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11th International Congress on Engineering and Food - Athens, Greece, 2011 i
ii
 FOOD PROCESS ENGINEERING
IN A CHANGING WORLD

Proceedings of the
11th International Congress on Engineering
and Food (ICEF11)

VOLUME III

Editors
Petros S. Taoukis
Nikolaos G. Stoforos
Vaios T. Karathanos
George D. Saravacos

ATHENS, GREECE
2011

11th International Congress on Engineering and Food - Athens, Greece, 2011 iii
Food Process Engineering in a Changing World
Proceedings of the 11th International Congress on Engineering and Food,
May 22-26, 2011, Athens, Greece.

ICEF 11 Secretariat:
Professor Petros Taoukis
School of Chemical Engineering
National Technical University of Athens
Athens 15780 Greece
e-mail: icef11@chemeng.ntua.gr

Technical & Scientific ICEF11 Editorial Team:

Dr. Efimia Dermesonluoglu, Dr. Eleni Gogou, Dr. Virginia Giannou, John
Tzigounakis
National Technical University of Athens

Published by:
Cosmosware, Ag. Ioannou 53, Athens, Greece, 0030 2106013922
cosmosware@ath.forthnet.gr

All papers appearing in the ICEF11 Proceedings were Peer Reviewed for acceptance by at least
two independent reviewers from the Scientific Committees.

Copyright © NTUA, School of Chemical Engineering, Athens 2011

SET ISBN: 978-960-89789-6-6
ISBN: 978-960-89789-5-9

iv
 ICEF11 ORGANIZERS

Organizer:
National Technical University of Athens, School of Chemical Engineering

Co-Organizers:
- Agricultural University of Athens
- Aristotle University of Thessaloniki
- Harokopion University of Athens
- Technical Chamber of Greece

Executive Committee:
President Prof. George Saravacos,
President of the International Association for Engineering and Food
Secretary Prof. Petros Taoukis, National Technical University of Athens
Treasurer Prof. Magda Krokida, National Technical University of Athens
Members Prof. Vaios Karathanos, Harokopion University of Athens
Prof. Harris Lazarides, Agricultural University of Thessaloniki
Prof. Nikolaos Stoforos, Agricultural University of Athens
Prof. Constantina Tzia, National Technical University of Athens
-Technical Chamber of Greece
Prof. Stavros Yanniotis, Agricultural University of Athens

International Honorary Committee:

J. Aguilera (Chile) P. Linko (Finland)
G.V. Barbosa-Canovas (USA) D. Lund (USA)
Z. Berk (Israel) B. McKenna (Ireland)
J.J. Bimbenet (France) A. Mujumdar (Singapore)
D. Farkas (USA) H. Schubert (Germany)
D.R. Heldman (USA) H. Schwartzberg (USA)
B. Hallstrom (Sweden) R.P. Singh (USA)
R. Jowitt (UK) W.E.L. Spiess (Germany)
M. Karel (USA) J. Welti-Chanes (Mexico)
D. Knorr (Germany) T. Yano (Japan)
T.P. Labuza (USA)

11th International Congress on Engineering and Food - Athens, Greece, 2011 v

vi
 ICEF11 SCIENTIFIC COMMITTEES

International Scientific Committee:

Abram, Veronika Dumoulin, Elisabeth Liao, Xiaojun
Afoakwa, Emmanuel Ohene Efremov, German Ivanovich Liapis, Athanasios
Aguilera, Jose, Miguel Erdogdu, Ferruh Lillford, Peter
Ahrne, Lilia Evans, Judith Lo, Martin Y.
Akterian, Stepan, G. Farkas, Brian E. Lund, Daryl
Alvarez, Graciela Fekete, Andras Marcotte, Michele
Alzamora, Stella Feng, Hao Mascheroni, Rodolfo H.
Bakalis, Serafim Fikiin, Kostadin Masi, Paolo
Balaban, Murat Fito, Pedro J. McCarthy, Michael
Balasubramaniam, V.M. (Bala) Floros, John D. McElhatton, Anna
Balla, Csaba Frias, Jesus M. McKenna, Brian
Barbosa-Canovas, Gustavo Fryer, Peter Miyawaki, Osato
Becker, Thomas Furuta, Takeshi Moresi, Mauro
Blahovec, Jiri Gekas, Vassilis Mujumdar, Arun Sadashiv
Boom, Remko Marcel Gonzalez-Martinez, Chelo Mulet, Antonio
Boudrant, Joseph Gutierrez-Lopez, Gustavo F. Murray, Andrew
Buckow, Roman Havet, Michel Nakanishi, Kazuhiro
Buera, Maria del Pilar Heinz, Volker Nedovic, Viktor
Chen, Hongda Heldman, Dennis R. Nguyen, Minh
Chernukha, Irina Hendrickx, Marc E.G. Nicolai, Bart
Chirife, Jorge Houska, Milan Niranjan, Keshavan
Choi, Yong-Hee Hubinger, Miriam Dupas O’Donnell, Colm
Ciprovica, Inga Hung, Yen-Con Okos, Martin
Clark, J. Peter Ives, Don Omobuwajo, Taiwo
Cleland, Don Karwe, Mukund Ortega-Rivas, Enrique
Costa, Rui Kaufmann, Stefan F.M. Ozilgen, Mustafa
Cunha, Luis Miguel Kaymak-Ertekin, Figen Ozkan, Necati
Cullen, Patrick J. Knorr, Dietrich Payne, Fred
Dalla Rosa, Marco Kokini, Jozef Pedreschi, Franco
Datta, Ashim Krijgsman, Ardjan Pham, Tuan
Davidson, Valerie J. Kristbergsson, Kristberg Pittia, Paola
De Carvalho, Rosemary Aparecida Labuza, Theodore Popa, Mona Elena
De Pinho, Samantha Cristina Laurindo, Joao Borges Poutanen, Kaisa
Dejmek, Petr Lebovka N.I., Nikolai Purwadaria, Hadi
Del Valle, Jose Manuel Lewicki, Piotr P. Rahman, Shafiur
Devahastin, Sakamon Li, Shu-Jun Ramaswamy, Hosahalli S.

11th International Congress on Engineering and Food - Athens, Greece, 2011 vii
Rao, Anandha (Andy)
Raspor, Peter
Razavi, Seyed M.A.
Reid, David
Roos, Yrjo
Sablani, Shyam
Saguy, Sam
Sastry, Sudhir K.
Schubert, Helmar
Shimoni, Eyal
Silva, Cristina L.M.
Simpson, Ricardo
National Scientific Committee:
Adamopoulos, Konstantinos G.
Bezirtzoglou, Eugenia
Biliaderis, Costas G.
Christakopoulos, Paul
Giannakourou, Maria
Goula, Athanasia
Karapantsios, Thodoris D.
Karathanos, Vaios
Kekos, Dimitris
Kiranoudis, Chris
Kolisis, Fragiskos
Kontominas, Michael
Kostaropoulos, Athanasios
viii
Singh, R. Paul
Singh, R.R.B.
Sobral, Paulo Jose do Amaral
Spiess, Walter E.L.
Sun, Da-Wen
Suzuki, Kanichi
Teixeira, Arthur A.
Tiwari, Brijesh
Torreggiani, Danila
Trystram, Gilles
Tsotsas, Evangelos
Tuzhilkin, Vyacheslav Ivanovitch
Koulouris, Alexandros
Koutinas, Apostolis
Koutsoumanis, Kostas
Krokida, Magda
Lambrinos, Grigoris
Lambropoulos, Athanasios
Lazarides, Harris
Lazos, Evangelos
Mallidis, Constantinos
Mandala, Ioanna
Marinos-Kouris, Dimitrios
Maroulis, Zacharias
Marousis, Stratis
Van de Voort, Frederick
Van der Linden, Erik
Van Impe, Jan
Venskutonis, Rimantas
Voilley, Andree
Vorobiev, Eugene
Welti-Chanes, Jorge
Windhab, Erich J.
Wu, James Swi-Bea
Zhou, Weibiao
Zwietering, Marcel
Nychas, George-John
Oreopoulou, Vassiliki
Papadakis, Spyridon
Raphaelides, Stylianos N.
Skandamis, Panagiotis
Stoforos, Nikolaos
Taoukis, Petros
Tzia, Constantina
Valdramidis, Vassilis
Xanthopoulos, George
Xenakis, Aristotelis
Yanniotis, Stavros
Zogzas, Nikolaos
 ICEF11 is organized under the auspices of the
International Association for Engineering and Food
(IAEF).

ICEF 11 Institutional Supporters

11th International Congress on Engineering and Food - Athens, Greece, 2011 ix

x
 ACKNOWLEDGEMENTS

The 11th International Congress on Engineering and Food has been supported by
the following companies and institutions:

Golden Sponsor
Delta

Silver Sponsors
Technical Chamber of Greece
Nestlé

Sponsors
Federation of Hellenic Food Industries (SEVT)
Coca-Cola 3E
Creta Farm
D. Nomikos
Papadopoulos
Elsevier
Hellenic Association of Chemical Engineers
Minerva S.A.
Sara Lee
Unilever
Agricultural Cooperatives Union Aeghion-Greece
Diversey Hellas
Gaea
INO

11th International Congress on Engineering and Food - Athens, Greece, 2011 xi

xii
 PREFACE

The present book is part of the three volume Proceedings of the 11th
International Congress on Engineering and Food (ICEF11) which took place in
Athens, Greece, May 22-26, 2011. The Congress was under the auspices of the
International Association for Engineering and Food (IAEF). It was organized by
the School of Chemical Engineering of the National Technical University of
Athens and the Agricultural University of Athens, the Aristotle University of
Thessaloniki, the Harokopion University of Athens and the Technical Chamber
of Greece.
Participation in ICEF 11 exceeded 1100 scientific papers presented by
colleagues from 70 countries. All papers were reviewed by at least two
independent members of an extended International Scientific Committee.
All submitted papers were assigned to 11 Scientific Topics: Food Materials
Science, Engineering Properties of Foods, Advances in Food Process Technology,
Novel Food Processes, Food Product Engineering & Functional Foods, Food
Waste Engineering, Hygienic Design and Operation of Food Plants, Modeling &
Control of Food Processes, Food Process Design & Economics, Modeling Food
Safety & Quality, and Innovation Management.
The 1st Volume contains the papers of the 415 Oral Presentations, while the 2nd
and 3rd Volumes contain the papers of the Poster Presentations. An
accompanying CD contains the extended electronic version of all papers included
in the three Volumes.
The Editors wish to thank the members of the Organizing and Scientific
Committees; their valuable contribution and help is greatly appreciated.

Petros Taoukis
Nikolaos Stoforos
Vaios Karathanos
George Saravacos

11th International Congress on Engineering and Food - Athens, Greece, 2011 xiii
xiv
 CONTENTS-VOLUME III

NOVEL FOOD PROCCESSES

Emerging technologies
Mass transfer of fruit slices in hypertonic solution 1609
F. A. Fazli, N. A. Fazli
Nanofiltration treatment of waste brine obtained from sugar decolorizing resin 1611
regeneration
F. Salehi, S.M.A. Razavi
Process development of ready-to-eat custard cream filled chinese steamed bun 1613
S. Chaiwanichsiri, N. Poonnakasem, K. Laohasongkram
Decontamination of spices by using a pulsed light treatment 1615
M. Moreau, I. Nicorescu, A.S. Turpin, A. Agoulon, S. Chevalier, N. Orange
Acceleration of precipitation formation in peach juice induced by high-pressure 1617
carbon dioxide
L. Zhou, Y. Zhang, X. Liaor, X. Hu
Effect of the electric field on the vitamins A, C and E alone and added to 1619
avocado paste
R.R.R. de la Torre, M.G.M. Ramos, Ma.R.R. López, J.A.A. Ortega, F.J.M. Montes
Effect of vacuum impregnation treatments to improve quality and texture of 1621
Zucchini (Cucurbita pepo, L.)
E. Occhino, I. Hernando, P. Pittia
Qualitative characteristics of sugar beet juices obtained in pilot extractor with 1623
pulsed electric field (PEF) pre-treatment
K. Loginova, E. Vorobiev, N. Lebovka
Modelling microbial load reduction in foods due to ozone impact 1625
E.M.C. Alexandre, T.R.S. Brandão, C.L.M. Silva
Use of organic acids on their own and in combination for decontamination of 1627
fresh vegetables and herbs as an alternative to chlorine
S. Bulut, E. OgrasÖcÖ
Use of a Weibullian model to characterize microbial inactivation in apple juice 1629
processed with ultraviolet light
E. Mytilinaki, S. Guerrero, S.M. Alzamora
Detection of pork freshness using NIR hyperspectral imaging 1631
D.F. Barbin, G. ElMasry, D.-W. Sun, P. Allen
Impact of non-thermal atmospheric pressure plasma on quality relevant food 1633
ingredients
B. Surowsky, F. Zülicke, O. Schlüter, D. Knorr

11th International Congress on Engineering and Food - Athens, Greece, 2011 xv

Effect of pulsed light and ascorbic acid/CaCl2 dipping on rheological properties 1635
of fresh-cut apples
P.L. Gómez, D.M. Salvatori, S.M. Alzamora
Modeling a pasteurization process of clarified apple juice based on pulsed 1637
ultraviolet light
I. Kasahara, P. Grogg, L. Aguilar
Encapsulation of Lactobacillus paracasei using Spray Gun technology 1639
M. Jiménez, E. Jiménez, E. Azuara, G. Luna, C.I. Beristain
Concentration of a vegetal enzymatic extract by microfiltration 1641
A.S.C. Teles, S.C. Terzi, L.F.M. Silva, F.S. Gomes, I.V.M. Moraes, A.S. Egito, L.M.C.
Cabral, V.M. Matta
Fresh produce decontamination by an atmospheric pressure plasma-jet 1643
M. Baier, M. Görgen, A. Fröhling, M. Geyer, W.B. Herppich, J. Ehlbeck, D. Knorr,
O. Schlüter
Intensification of process of water-thermal treatment of wheat grain before 1645
bread flour milling
O. Safonova, O. Razborskaya, V. Yuferov, O. Ozerov
The effect of abiotic stress pre-treatments on quality attributes of fresh-cut 1647
carrot cv. Nantes
C. Alegria, J. Pinheiro, M. Duthoit, E.M. Gonçalves, M.T. Coelho, M. Moldão-Martins,
M. Abreu
Yogurt from ultrasound treated milk: monitoring of fermentation process and 1649
evaluation of product quality characteristics
P. Sfakianakis, C. Tzia
Effect of sonication on malting behaviour of barley 1651
E. Dutheil, B. Tiwari, M. Gupta, P.J. Cullen, C. Brennan, C. O'Donnell

High pressure processing

A mathematical approach for using multiple enzyme based pressure- 1653
temperature-time integrators (PTTIs) for high pressure process evaluation
E. Gogou, P.Taoukis
Effect of high hydrostatic pressure treatments on physicochemical properties, 1655
microbial quality and sensory attributes of beef carpaccio
N. Szerman, Y. Barrio, B. Schroeder, P. Martinez, A. Sancho, C. Sanow, S.R. Vaudagna
Rheological properties of high pressure milk cream 1657
G. Donsì, G. Ferrari, P. Maresca
Effects of HHP combined with blanching on microorganisms and qualities of 1659
cloudy and clear strawberry juices
X. Cao, Y. Zhang, X. Liao, X. Hu
Effect of high pressure homogenization process on Bacillus stearothermophilus 1661
and Clostridium sporogenes spores in skim milk
C.R.G. Pinho, M.A. Franchi, A.A.L. Tribst, M. Cristianini

xvi
Effect of ultra high pressure homogenization on alkaline phosphatase and 1663
lactoperoxidase activity in raw skim milk
C.R. G. Pinho, M.A. Franchi, A.A.L. Tribst, M. Cristianini
Changes in texture, structure and pectin of peach during pressurization, heating 1665
or processing of high-pressure-induced and heat-induced jam
H. Kuwada, Y. Jibu, K. Nakamura, M. Tabuchi, Ai. Teramoto, K. Ishii, Y. Kimura, M.
Fuchigami
Effects of high pressure with the addition of sugar-alcohol on the improvement 1667
in texture and structure of frozen egg custard gel
A. Teramoto, Y. Jibu, H. Kuwada, Y. Kimura, K. Ishii, M. Fuchigami
Process variables study on supercritical CO2 extraction of Brazilian cherry seeds 1669
(Eugenia uniflora L.) rich in bioactive volatile
D. Nascimento e Santos, L.L. de Souza, N.J. Ferreira, A.L. de Oliveira
High hydrostatic pressure (HHP) microbial kinetics in orange comminuted 1671
V. Serment-Moreno, Z. Escobedo-Avellaneda, J. Welti-Chanes
Research development of ultra-high pressure processing on fruit juice 1673
W. Han, Z. Yunchuan, H. Qinghua, Z. Youbin
Effects of high hydrostatic pressure on antioxidant activity, mineral and starch 1675
content and bioaccessibility, in apple (Granny smith)
V. Briones-Labarca, G. Venegas-Cubillos, S. Ortiz-Portilla, M. Chacana-Ojeda, H.
Maureira
Microbiological stabilization of Aloe vera (Aloe barbadensis Miller) gel by high 1677
hydrostatic pressure treatments
J.E. Reyes, G. Tabilo-Munizaga, M. Guanoquiza, A. Vega-Galvez, M. Miranda, M.
Pérez-Won
Establishment of a processing method for tofu using high pressure compared to 1679
the heat induced method
Y. Jibu, K. Nakamura, A. Teramoto, H. Kuwada, M. Fuchigami
Enhanced infusion under High Pressure: New insights 1681
S. Mahadevan, M.V. Karwe
Structural changes of pectin methylesterase from orange peel subjected to 1683
thermal and high pressure processing
Z. Alexandrakis, T. Papadopoulos, F. Stavros, G. Katsaros, P. Katapodis, G. Nounesis,
P. Taoukis
Innovative value propositions for the food industry through non-thermal 1685
processing techniques
F. Purroy, C. Tonello

Separation and purification processes
Fractionation of liquid egg yolk: Influence of chemical and structural 1687
characteristics of egg yolk granular and plasma fraction on the continuous
centrifugal separation process
T. Strixner, M. Betz, U. Kulozik

11th International Congress on Engineering and Food - Athens, Greece, 2011 xvii
xviii
Bioprocessing engineering
Characterization of novel cholesterol esterase from Trichoderma sp. AS59 with 1721
high ability to synthesize steryl esters
A. Maeda, N. Hashitani, T. Mizuno, M. Bunya
Recovery of an antibacterial peptide fraction from snow crab by-products 1723
hydrolysate by electrodialysis with ultrafiltration membranes
A. Doyen, L. Saucier, L. Beaulieu, Y. Pouliot, M. Araya-Farias, L. Bazinet
Prospection of bacterial endophytes isolated from Baru (Dipteryx alata Vog.) 1725
as a potential source of bioactive compounds
G. Molina, A.P. Dionísio, M.R. Pimentel, G.T. Makita, R.C. dos Reis, G.M. Pastore
Biotransformation of R-(+)- and S-(ಥ)-limonene by Fusarium oxysporum 1727
G. Molina, R.L. da Costa, A.P. Dionísio, J.L. Bicas, G.M. Pastore

Novel food processes

Pulsed light decontamination of vegetables and fruits 1729
G. Pataro, G. Donsì, G. Ferrari
Shelf life extension of fresh-cut fruit by UV-light exposure 1731
L. Manzocco, S. Da Pieve, I. Bartolomeoli, M. Maifreni
Effect of ozonation on the sensory characteristics and pasting properties of 1733
cassava starch
E.O.C. Amorim, V.C. Doval, M. Cristianini
Production of antioxidant enriched cranberry juice by electrodialysis with 1735
filtration membrane: impact of process on juice composition
L. Bazinet, S. Brianceau, M. Araya-Farias, Y. Desjardins
Effect of sunflower oil applied by vacuum impregnation to refrigerated atlantic 1737
salmo
L. Puente, J. Ortiz, M. Leiva, S. Aubourg
Production of Mucor griceocyanus protease using different carbon sources in 1739
submerged fermentation
A. Ramírez, J. Sánchez, A. Iliná, J.C. Dusted Mendoza, J. Rodríguez, J.L. Martínez
Evaluation of MAP design parameters on quality of fresh-cut produce
F. Oliveira, M. Sousa-Gallagher, P. Mahajan, J. Teixeira 1741
Rational method for designing efficient food separation processes by 1743
chromatography. “Polyphenol-ethanol/water system with polymer-resins”
M. Hosono, R. Maeda, N. Yoshimoto, S. Yamamoto
Food-grade emulsions prepared by membrane emulsification techniques 1745
F. Spyropoulos, R.D. Hancocks, I.T. Norton
Use of supercritical CO2 for the inactivation of Aspergillus niger inoculated on 1747
stainless steel plates surface
M.A. da Silva, J. de Souza Ferreira, B.T. Iamanaka, F.S. Kihara, R.S. Cutolo, T.G.
Kieckbusch

11th International Congress on Engineering and Food - Athens, Greece, 2011 xix
Non-aqueous thermal processing of foods 1749
R. Steele, C. Kerjean

MODELING FOOD SAFETY & QUALITY

New technologies for the evaluation of quality and safety
Estimation of peroxidase activity in red cabbage by artificial neural network 1751
(ANN)
I. Shahabi Ghahfarrokhi, A. Daraei Garmakhany, S.M. Mousavi
Quality classification of corn tortillas by means of cross validation between 1753
sensorial evaluation and computer vision system
J.J. Chanona-Pérez, D. Mery, A. Soto, J.M. Aguilera, A. Cipriano, N. Veléz-Rivera, I.
Arzate-Vázquez, G.F. Gutiérrez-López
Effect of microwave blanching on acrylamide content and quality attributes of 1755
french fries
S. Tuta, K. Palazoglu, V. Gökmen
Effects of application of tranglutaminase in wheat proteins during the 1757
production of bread
E.Ap. Guastaferro Seravalli, A. Miwa Iguti, I.Ap. Santana, F. Finardi Filho
Agrophysical methods to determine bioenergetic status of agricultural 1759
products
J. Horabik, P. Baranowski, J. Tys
Separation between high and low quality coffees by FTIR-ATR 1761
A.S. Franca, A.P. Craig, L.S. Oliveira
Effect of temperature on biospeckle activity in apples 1763
A. Kurenda, A. Adamiak, A. Zdunek
Implementation of DNA technology in a Greek dairy company: An overview 1765
E. Beletsiotis, D. Ghikas, K. Kalantzi
Sensorial characteristics of goat milk cheeses made from ultra high-pressure 1767
homogenization-treated milk
B. Juan, J.M. Quevedo, B. Guamis, V. Ferragut, A.J. Trujillo
User-friendly software predicting the microbial spoilage of emulsified acid 1769
foods
S.G. Manios, A. Psomas, P.N. Skandamis
Detection of fecal contamination on leafy greens by hyperspectral imaging 1771
S. Kang, K. Lee, J.-G. Lim, M.S. Kim
Detection of mushroom Virus X (MVX) infection in asymptomatic 1773
mushrooms using FTIR microscopic imaging
L. Alvarez-Jubete, F. Bonnier, H. Byrne, H. Grogan, J.M. Frias
Design and validation of sensory focused processes of foods 1775
C. Tzia, V. Giannou, D. Lebesi, D. Sabanis, V. Polychniatou, P. Sfakianakis, C.
Chranioti, P. Moutsatsou

xx
Rapid HPTLC-based method for quality control: simultaneous chemical 1777
analysis and antioxidant activity determination in herbal, nutraceutical and
functional foods
K. Muñoz, J. Calderón, E. Osorio, D. Castro, R. Serna, J. Díaz, J. Londoño
Nondestructive evaluation of watermelon ripeness using LDV 1779
R. Abbaszadeh, A. Rajabipour, H. Ahmadi, M. Mahjoob, M. Delshad
Effect of pasteurization on bioactive amines in human milk 1781
F.F. Silva, M.B.A. Gloria
Integration of new/rapid methods and ICTs to improve food safety and 1783
quality
D. Lebesi, A. Bilbao, A.I. Díaz, I. Papadaki, V. Oreopoulou

Modeling of quality and safety and predictive microbiology

Commercial characterization of Madalenas: Relationship between physical 1785
and sensory parameters
M.M. Ureta, D.F. Olivera, V.O. Salvadori
Integrating strain variability in modelling Salmonella enterica growth 1787
A. Lianou, K. Koutsoumanis
A study on germination time and mycelium growth kinetics of single fungal 1788
spores
M. Gougouli, K. Koutsoumanis
Quantifying the combined effect of salt and temperature on the growth of 1789
Listeria strains isolated from salmon and salmon processing environments
T. Skåra, A.M Cappuyns, E. Van Derlinden, J.T. Rosnes, V.P. Valdramidis, J.F.M.
Van Impe
Modelling thermosonication inactivation of Aspergillus flavus combining 1791
natural antimicrobial at different pH
C.P. Coronel, M.T. Jiménez, A. López-Malo, E. Palou
Survival of Bifidobacterium longum in model solutions and fruit juices 1793
S. Nualkaekul, I. Salmeron, D. Charalampopoulos
Inactivation kinetics of attached Escherichia coli cells on stainless steel and 1795
fresh-cut apples by hydrogen peroxide disinfection treatments
S. Raffellini, S. Ortiz, S.N. Guerrero, S.M. Alzamora
Bi-phasic growth of Listeria monocytogenes Scott A in Modified 1797
Welshimer’s broth at 7, 10 and 14°C
N.A. Tyrovouzis, A.S. Angelidis, N.G. Stoforos

Reaction kinetics in food processing

Kinetic of white chocolate color loss 1799
D.C.P. Jardim, AG. Orse, P. Efraim, S.C.S.R. de Moura
Available lysine in powdered infant formula as described by reaction kinetics 1801
I. Schmitz, A. Gianfrancesco, U. Kulozik, P. Foerst

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxi
Kinetic modelling of colour changes during beef roasting 1803
S.M. Goñi, V.O. Salvadori
Instrumentation of a semi-industrial oven to monitor non-enzymatic 1805
browning kinetics during baking
M. Courel, B. Rega, S. Fehaili, P. Giampaoli, C. Bonazzi

Degradation of 5-hydroxymethylfurfural in malt during fermentation of beer 1807

G. AkÖllÖoølu, B. Ataç Mogol, V. Gökmen
Thermal inactivation kinetics of L-carnitine 1809
P. Prokopiou, A.M. Goula, N.G. Stoforos
Quality degradation of butterhead lettuce: the performance of General 1811
Stability Index (GSI) modified methodology
M.V. Agüero, S.I. Roura
A MALST method comparison over univariate kinetic modelling for 1813
determination of Shelf life in cereal snack of dried apples
J. Saavedra, A. Córdova, C. Quezada
Modulation of thermal inactivation of protease during enzymatic hydrolysis 1815
of salmon muscle
P. Valencia, N. Bustos, S. Almonacid

Risk assessment and safety assurance

Determination of aflatoxin M1 in raw milk by HPLC marker as evidence of 1817
cattle-food storage conditions from the herd suppliers of a dairy company in
the city of Valledupar
E. Fragoso, T. David, S. Romero, H. Ospino
Use of a Poisson-gamma regression model to assess the process hygiene 1819
criterion for Enterobacteriaceae on Irish sheep carcasses
U. Gonzales-Barron, F. Butler
Improvement of harvesting and processing of cultivated fresh water prawn 1821
(Macrobrachium rosenbergii)
T.C.A. Silva, L.S. Arrieche
Assessing the conditions of milk production on farms based on family farming 1823
M. da Penha Piccolo Ramos, F.C.N.N.Silva, L. Oliveira de Fariña, C. L. de Oliveira
Pinto
Regeneration of frying oils by using adsorbent resins 1825
N. Göncüoølu, B.Atac Mogol, V. Gökmen
Extending shelf life of watercress by means of alternative sanitizers and 1827
modified atmosphere packaging
C. Char, P. Villena, A. Hinojosa, V. Escalona
Modeling the effect of acid and osmotic shifts above and across the growth 1829
boundaries on the adaptation and growth of Listeria monocytogenes
C.-I. A. Belessi, S.I. Merkouri, A.S. Gounadaki, S. Schvartzman, K. Jordan, E.H.
Drosinos, P.N. Skandamis

xxii
Effect of contamination stage and inoculum history on the survival and 1831
growth of Listeria monocytogenes in semi-hard and hard cheese
C.-I.A. Belessi, S. Arapaki, A.S. Gounadaki, P.N. Skandamis
Inoculated pack study of an intermediate moisture egg patty 1833
M. Richardson, A. Sikes, C. Lee, S. Walker
HACCP implementation in public hospitals: a survey in Crete, Greece 1835
E. Kokkinakis, A. Kokkinaki, G. Kyriakidis, A. Markaki, G.A. Fragkiadakis
HACCP implementation in local food industry: a survey in Crete, Greece 1837
E. Kokkinakis, A. Kokkinaki, G. Kyriakidis, A. Markaki, G.A. Fragkiadakis

Management and optimization of the food chain-from production to

consumption
A simplified method for determination of the sour cassava starch expansion 1839
property
M. Janete Angeloni Marcon, D. Jacob Kurtz, M. Maraschin, V. Reginatto, I. Mottin
Demiate, E.R. Amante
Influence of room temperature on food safety in refrigerated display cabinet 1841
O. Laguerre, M. Hoang, G. Alvarez, D. Flick
Antemortem and postmortem biochemistry, drip loss and lipid oxidation of 1843
European sea bass muscle tissue
C. Nathanailides, S. Panopoulos, F. Kakali, C. Karipoglou, D. Lenas
Impact of initial handling and subsequent storage conditions on the safety 1845
and keeping quality of sardines
K. Chatzikyriakidou, E. Katsanidis
Survival of Salmonella and Escherichia coli O157:H7 during freezing, 1847
thawing and cooking of ground beef patties, simulating common household
practises
S.G. Manios, T. Giovanis, A. Lalechou, P.N. Skandamis
European food, technology and nutrition declaration (EFTN Declaration) 1849
P. Raspor, L. Baša
Optimization of shelf life distribution of frozen fish products based on 1851
modelling and TTI monitoring
M.N. Giannoglou , M. Loukianou, K. Tsatsaragou, T. Tsironi, P.S. Taoukis

Modeling food safety and quality

Modeling of Greek coffee aroma loss during storage at different temperatures 1853
and water activities
E. Makri, D. Tsimogiannis, E. Dermesonluoglu, P. Taoukis
Combined effect of meat composition and heating parameters on the 1855
physicochemical state of proteins
A. Promeyrat, L. Le Louët, A. Kondjoyan, T. Astruc, V. Santé-Lhoutellier, P. Gatellier,
J.D. Daudin

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxiii
Biogenic amine levels in dry fermented sausages produced and sold in Greece 1857
E.J. Papavergou
Spore inactivation by ultraviolet irradiation combining with different pre- 1859
heating treatment
D. Hamanaka, H. Yamada, T. Kadoyanagi, V. Tryvittayasil, F. Tanaka, T. Uchino
Aroma profile of different salted dried codfishes 1861
M. Costa Silva, L.R. Silva, P. Guedes-de-Pinho, P. Andrade, P. Valentão, R. Costa
Influences of pH and temperature on infrared spectroscopic features of brewed 1863
coffee
A. Hashimoto, Y. Sugimoto, K.-I. Suehara, T. Kameoka
Comparison of wild and farmed sea bass (Dicentrarchus labrax L) lipid quality 1865
D. Lenas, S. Chatziantoniou, C. Nathanailides, D. Triantafillou
Coupling between heat and mass transfer and stoechio-kinetic models to bring 1867
insight into maillard reaction kinetics during baking of sponge-cake products
C. Pénicaud, B. Broyart, D. Goujot, M. Courel, X.-M. Meyer, C. Bonazzi
A methodology for the certification of food-serving services according to the 1869
Mediterranean dietary model
E. Grigoroudis, A. Psaroudaki
Bactericidal effect of electrolyzed oxidizing (EO) water on E. coli O157:H7- 1871
and Salmonella-inoculated beef, chicken, and shrimp
J. Weese, T.-S. Huang
Predicting persimmon puree colour as a result of puree strength manipulation 1873
A.R. East, X.H. Tan, J. Suntudprom
Occurrence of furan in commercial samples of roasted coffee in Brazil 1875
A.P. Arisseto, E. Vicente, M.S. Ueno, M.C.F. Toledo
Potential of furan formation in roasted coffee as influenced by species and roast 1877
degree
A.P. Arisseto, E. Vicente, M.S. Ueno, S.A.V. Tfouni, M.C.F. Toledo
Thermal inactivation of Byssochlamys nivea in pineapple juice combined with 1879
preliminary high pressure treatments
E.H. da Rocha Ferreira, A. Rosenthal, V. Calado, J. Saraiva, S. Mendo, P. Rodrigues
De Massaguer
Role of spices on acrylamide formation in buckwheat ginger cakes 1881
L. Marková, Z. Ciesarová, K. Kukurová, H. Zieliľski, D. Zieliľska, A. Bednáriková
Detection of deoxynivalenol in wheat flour using fluorescence fingerprint 1883
J. Sugiyama, K. Fujita, M. Tsuta, M. Kushiro
Modeling of growth and ochratoxin A production of Aspergillus carbonarius 1885
and evaluation in food matrices: effect of (gel) microstructure, water activity,
and temperature
A.E. Kapetanakou, A. Abavi, S. Yanniotis, E.H. Drosinos, P.N. Skandamis
Modelling of in-mouth perception the case of sodium 1887
B.J.D. Le Révérend, I.T. Norton, S. Bakalis

xxiv
Furan derivatives dynamic in rye bread processing 1889
V. Ozolina, D. Kunkulberga, B. Cieslak, M. Obiedzinski
The effects of ƈeracleum platytaenium boiss essential oil on the growth of
ochratoxigenic penicillium verrucosum (D-99756) isolated from kashar cheese 1891
S. Ozcakmak, M. Dervisoglu, A. Akgun, A. Akcin, T. Aytaû Akcin, F. Seyis
The inhibition of contaminated molds by some essential oils in cheeses 1893
S. Ozcakmak, A. Akgun, M. Dervisoglu
Fungicidal against Aspergillus flavus and decontaminate Aflatoxin B1 with 1895
Neutralized and Acidic electrolyzed oxidizing water
Li Lite, Xiong Ke


ADVANCES IN FOOD PROCESS TECHNOLOGY

Cooling and freezing
Influence of different inulin types on bread quality in the process of freezing and 1897
thawing
J.S. Filipoviý, đ.B. Psodorov, N.K. Filipoviý, V.S. Filipoviý
Thermal analysis of strawberry preservation by cooling and freezing 1899
A.-G. Ghiaus, C. Vasilescu
Effects on Xe hydrate formation for texture in vegetable tissue 1901
H. Ando, T. Suzuki, K. Kajiwara, Y. Kawagoe, Y. Makino, S. Oshita
The potential of ambient cooling systems for reducing refrigeration loads and 1903
saving energy
S.J. James, C. James
Industrial superchilling, a practical approach 1905
A.M. Stevik, I.C. Claussen

Thermal processing
Evaluation of thermal resistance and efficiency of palm olein and canola oils in 1907
frying of potato chips
A. Rafe, S. Bolourian, G. Goli Movahhed, M. Afshari
Assessment of furfurol derivatives: food risk factors in natural apricot and peach 1909
juice
C. Jianu, I. Cocan, I. Jianu
Numerical evaluation of liquid food heat sterilization in a brick-shaped package 1911
P.E.D. Augusto, M. Cristianini
Effect of steam jet cooking on the destruction of corn starches 1913
L.H. Ferng, S.H. Chen, Y.A. Lin
Experimental studies and interpretation of pistachio nut roasting process 1915
G. Trystram, R. Yeganeh

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxv
Heat transfer analysis-based prediction of protein denaturation and umami 1917
component of meat during cooking
N. Ishiwatari, M. Fukuoka, N. Hamada, N. Sakai
Effect of steam cooking of food on mass transfer 1919
E. Descours, E. Ferret, N. Valance, A. Voilley, A.-M. Seuvre
Development of experimental devices in order to study the interactions between 1921
heat and mass phenomena and thermal degradation reactions of lipids during
domestic reheating of pre-fried food products
J. Cernela, ƃ. Heyd, ƃ. Broyart
The effect of UHT and VAT thermal processing systems on whey protein 1923
denaturation and gel strength of yoghurt
A. Labropoulos, T. Varzakas, S. Anestis
Application of ohmic heating to whole egg 1925
T. Nakai, M. Fukuoka, N. Sakai
Transient mass and heat transfer during potato deep fat frying - The effect of 1927
the oil type, frying load and initial frying temperature
J.S. Lioumbas, M. Kostoglou, T.D. Karapantsios


Innovation in traditional processing
Acceptance of Iron Fortified Rice (I-Rice) in the Philippines to Combat Iron 1929
Deficiency Anemia (IDA)
E.M. San Juan, N.O. Camitan, A.C. Natividad, M.U. Gochangco, L.D. Alkuino, A.R.
Cariso, Jr., A.O. Lustre, A.W. Tejada
Quality characteristics and drying behaviour of muffins baked in steam assisted 1931
and convectional ovens
M. Sakin Yilmazer, H. Isleroglu, T. Kemerli, O. Ozdestan, G. Guven, F. Kaymak-
Ertekin, A. Uren, B. Ozyurt
Study of an innovative combination between microwaves and enzymes applied to 1933
bakery products
T. De Pilli, A. Derossi, R. Giuliani, C. Severini
Effective removal of heavy metal in some fish sauce products by tannin treatment 1935
T. Sasaki, T. Michihata, S. Nakamura, T. Enomoto, T. Koyanagi, H. Taniguchi, M.
Aburatani, M Koudou, K. Tokuda
Crispy air-dried pineapple rings: optimization of processing parameters 1937
G. Cortellino, P. Pani, D. Torreggiani
Extraction of polyphenols from grape seeds by unconventional methods and 1939
extract concentration through polymeric membrane
D. Liu, E. Vorobiev, R. Savoire, J.-L. Lanoisellé
Performance of bovine and ovine liquid whey protein concentrate on functional 1941
properties of set yoghurts
M. Henriques, D. Gomes, D. Rodrigues, C. Pereira, M. Gil

xxvi
Manufacture of gelatin-based films using extrusion: Assessment of extrusion 1943
parameters on film properties
Z.A. Nur Hanani, E. Beatty, Y.H. Roos, J.P. Kerry
Combining microwave and jet-impingement in a oven prototype 1945
G. Ruocco, M.V. De Bonis, F. Marra
The sequential ventilation of cheese ripening rooms: an eco-design approach? 1947
P.-S. Mirade, B. Perret, H. Guillemin, D. Picque, C. Callon, M.-C. Montel, G. Corrieu
Influence of additives on white tin loaf alveolloli formation 1949
C. Domingues, P. Prazeres, P. Correia
Textural properties of vegetables: a key parameter on ultrasonic assisted 1951
convective drying
C. Ozuna, J.A. Cárcel, J.V. Santacatalina, A. Mulet, J.V. García-Pérez
The influence of palm oil quality on the refining conditions 1953
K.A. Sampaio, J.V. Ayalla, S.M. Silva, R. Ceriani, R. Verhé, A.J.A. Meirelles
Challenges and solutions of a novel muscle-food processing technology: acid and 1955
alkaline solubilization
P.K. Vareltzis, K.G. Adamopoulos, H.O. Hultin†
Effect of various proteins on characteristics and synerisis of tzatziki 1957
A.G. Stefanakis, E.K. Stavrakakis, K.G. Adamopoulos, P.K. Vareltzis, A.M. Goula
High-power ultrasound-assisted pasteurisation of honey 1959
D. Kabbani, F. Sepulcre, J. Wedekind, E. Gaston
Fourier transform infrared (FTIR) spectroscopic analysis of biodegradable 1961
gelatin films immersed in water
Z.A. Nur Hanani, Y.H. Roos, J.P. Kerry
Effects of edible chitosan- linseed mucilage coating on quality and shelf life of 1963
fresh-cut strawberry
L.E. Pérez Cabrera, G.C. Díaz Narváez, A. Tecante Coronel, C. González Martínez
How to apply acrylamide mitigation tools in food technology 1965
Z. Ciesarová, K. Kukurova, L. Markova, J. Sadecka
Coconut water processing using ultrafiltration and pasteurization 1967
L.A. Nakano, W.F. Leal Jr., D.G.C. Freitas, L.M.C. Cabral, E.M. Penha, A.L. Penteado,
V.M. Matta

Concentration and dehydration processes
Progressive freeze-concentration: Improvement and applications 1969
O. Miyawaki
Study of color and shrinkage of Physalis peruviana during convective drying by 1971
computer vision
L. Puente, C. Pinto, E. Echegaray, E. Castro, M. Cortés
Optimization of osmotic dehydration process coupled with ohmic heating using 1973
granny smith apples
A. Sepúlveda, S. Sastry, J. Moreno, H. Nuñez, S. Almonacid, R. Simpson

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxvii
Analysis of the quality attributes of osmotically dehydrated mango 1975
M.A. Khan, L. Ahrné, J. Oliveira
Osmotic dehydration process coupled with ohmic heating using granny smith 1977
apples and its effects on product quality
R. Simpson, C. Farias, V. Medina, S. Almonacid, H. Nuñez
Physico-chemical, rheological and sensory properties of shamia date sheets 1979
K. Youssef, A. Shatta, T. Moussa-Ayoub, S. El-Samahy
Colour stability of spinach leaves during freeze processing steps 1981
K. Youssef, A. Shatta, A, Al-Sanabani, S, El-Samahy
Study on combined hot-air and microwave vacuum drying for scallion 1983
Y. Li, S. Li, B. Yang, Q. Han, J. Ma, D. Zhao
Experimental study of vacuum discharge in microwave freeze-drying process 1985
Y. Cao, S. Li, B. Yang, F. Zhao, D. Su, Q. Zhao
Ultrasound application as pre-treatment for drying of fruits 1987
F.A.N. Fernandes, S. Rodrigues
A basic investigation on instant coffee production by vacuum belt drying 1989
K. Burmester, A. Pietsch, R. Eggers
Shrinkage of papaya (Carica papaya L.) during convctive drying: Influence of 1991
glass transition phenomenon
L.E. Kurozawa, M.D. Hubinger, K.J. Park
Influence of sucrose replacement on colour and texture of kiwi jam 1993
E. Rosa, I. Peinado, A. Heredia, A. Andrés
Influence of dry and wet osmotic dehydration on colour and texture of a spread 1995
kiwi product
I. Peinado, E. Rosa, A. Heredia, A. Andrés
Quality assessment of dried eggplant using different drying methods: hot air 1997
drying, vacuum freeze drying and atmospheric freeze drying
J.V. Santacatalina, C. Ozuna, J.A. Cárcel, J.V. García-Pérez, A. Mulet
Effect of Fluidized-bed drying on the microstructure of higuerilla seeds (Ricinus 1999
communis). An alternative source of protein and biofuel
J.J. Chanona Pérez, E. Terrés Rojas, J.A. Mendoza Pérez, H.M. Hernández, G.F.
Gutiérrez López, V. Garibay Febles, M. de Jesús Perea Flores
A simple mathematical model proposed to predict kinetics of mass transfer in 2001
osmotic dehydration of muskmelon
J. Lucena Barbosa, D.G. Correa Moreira Rocha, M.I. Martins Jacintho Barbosa, M.
Cordeiro Mancini, M. Dupas Hubinger
Drying characteristics of Açaí (Euterpe oleracea) 2003
A.M. Barbosa Neto, L.G. Marques, M.M. Prado
Vitamin C content of freeze-dried tropical fruits 2005
L.G. Marques, M.M. Prado, J.T. Freire
Rehydration characteristics of freeze-dried avocado (Persea americana) 2007
D.S. Souza, J.D.R. Pimentel, M.M. Prado, L.G. Marques, N. Narain


xxviii
Mechanical processing of foods
Studies on the cooking conditions and mechanical koji-making of black beans 2009
C.-L. Jao, W.-C. Ko, K.-C. Hsu
The use of xylanase to improve physicochemical characteristics of 2011
nixtamalized corn flour and tortilla texture obtained by extrusion
B. Ramírez-Wong, L.C. Platt-Lucero, P.I. Torres-Chávez, J. López-Cervantes, D.I.
Sánchez-Machado, E. Carvajal-Millán, F. Martínez-Bustos, A. Quintero-Ramos, I.
Morales Rosas
The design of non-contact automatic shell cutting machine of chestnut and the 2013
investigation of its effect by means of chestnut shelling experiment
H.-W. Xiao, Z.-L. Du, Z. Lou, L.-H. Wang, J.-W. Bai, Z.-J. Gao
Relationship between chromatographic profiling by HS-SPME and sensory 2015
quality of mandarin juices: effect of squeeze technology
R. Alvarez Quintero, C. Passaro Carvalho, O. Lara Guzmán, J. Londono Londoño

Advances in Food Processing Technologies

Effect of magnetic fields and ultrasound on aerobic mesophiles and histamine in 2017
beef loin tuna loin tuna (Thunnus albacares)
V.M. Gélvez Ordóñez, L. Fuentes Berrio
Relationship between pectic substances and strand separation of cooked 2019
spaghetti squash
K. Ishii, A. Teramoto, H. Kuwada, Y. Jibu, M. Tabuchi, Y. Kimura, M. Fuchigami
Improvement of an enzymatic process to elaborate orange segments in syrup 2021
R. Robles-López, A. Dorantes-Nieto, D. Díaz-Carvajal, R.R. Robles-De la Torre, M.D.
Bibbins-Martínez
The technology of butters’ enriching with carrots’ powder 2023
ǟ.Ǜ. Rashevska, Ǜ.Ǚ. Vasheka
Production of ewe’s milk cheese using calf rennet and a plant coagulant from 2025
flowers of cardoon Cynara cardunculus: Proteolysis during ripening
J. Fernández-Salguero, A. Pino, E. Galán
Production of ewe’s milk cheese using calf rennet and a plant coagulant from 2027
flowers of cardoon Cynara cardunculus: Sensory characteristics during ripening
E. Galán, R. González, J. Fernández-Salguero
Functional drink production through pomegranate juice fermentation 2029
S. Plessas, M. Koulis, A. Alexopoulos, E. Bezirtzoglou








11th International Congress on Engineering and Food - Athens, Greece, 2011 xxix
xxx
11th International Congress on Engineering and Food - Athens, Greece, 2011 xxxi
Functional foods enriched in aloe vera. Effects of vacuum impregnation and 2089
temperature on the respiration rate and the respiratory quotient of some
vegetables
S. Sanzana, M.L. Gras, D. Vidal-Brotóns
Production of 4th range Iceberg lettuce enriched with calcium. Evaluation of 2091
some quality parameters
M. L. Gras, D. Vidal-Brotóns, F.A. Vásquez-Forttes
Microencapsulation of probiotic bacteria with alginate and prebiotic and 2093
evaluation of survival in ice cream
C. Jurkiewicz, M.P.M. Boscarioli, R.G. Ferreira, E.P. Ribeiro, L. Kunigk
The influence of operational parameters in the pectin agglomeration 2095
T.A. Medeiros Hirata, V. Goulart Machado, G. César Dacanal, F.C. Menegalli
Antioxidant activity of microcapsules of Rubus sp. juice using spray drying 2097
M. Jimenez, E. Azuara, J. Vernon-Carter, G. Luna-Solano, C.I. Beristain
Novel ways to control enzymatic hydrolysis as a tool to produce functional 2099
peptides
E. Leeb, U. Kulozik, S. Cheison
Influence of the structure and composition of the País grape proanthocyanidins 2101
on the inhibition of angiotensin converting enzyme
S. Godoy, M. Roeckel, E. Aspé, K. Fernández
Kinetic characterization of inhibition of angiotensin converting enzyme by 2103
proanthocyanidins extracted from vitis vinífera L. cv. País
K. Álvarez, M. Roeckel, E. Aspé, K. Fernández
Enzymatic depolymerisation of oat Ƣ-glucan 2105
A.-I. Ninios, J. Sibakov, I. Mandala, K. Fasseas, K. Poutanen, E. Nordlund, P. Lehtinen
Parameters evaluation of fructooligosaccharides production by sucrose 2107
biotransformation using an osmophilic Aureobasium pullulans strain
J. Bueno da Silva, A.E. Cavalcante Fai, R. dos Santos, L.C. Basso, G.M. Pastore
Obtaining and characterization of mango peel powder and its use as a source of 2109
fiber and a functional ingredient in natural yogurt
C. Ruiz, C. Ramírez, C. Gutiérrez de Piñeresc, M. Ángulo, J. Hedreira
Influence of gamma radiation on sprouting inhibition of the rhizomes and 2111
on the quality of turmeric
L. Peret-Almeida, M.B.A. Gloria

Food product development

Antioxidant dyes and pigment extraction using a home-made pressurized 2113
solvent extraction system
D.T. Santos, C.L.C. Albuquerque, M.A.A. Meireles
Comparative study of the physicochemical characteristics of an economic 2115
Buffalo (Bubalus bubalis) meat product and an economic beef (Bos indicus)
meat product with incorporation of bovine hemoglobin in powder in both
formulations
J.F. Rey, C.L. Martínez, A. Urrea

xxxii
Production of Turkish delight (lokum) with its additives and quality 2117
A. Batu
Effect of fermented okara (bean curd lees) intake on TNCB (2, 4, 6- 2119
trinitrochlorobenzene)-induced chronic dermatitis in NC/Nga mice
T. Enomoto, M. Nishi, F. Barla, N. Murata, H. Matsui, H. Kumagai, H. Take, T.
Michihata, S. Nakamura, M. Kawashima, E. Fujihara
Development of a dehydrated and laminated probiotic product with B. 2121
infantis and L. acidophilus using goat sweet whey
G. Trujillo de Santiago, C. Sáenz Collins, C. Rojas de Gante
Lentil-based snacks: Structural and textural evaluation 2123
A. Lazou, M. Krokida, N. Zogzas, V. Karathanos
The study on SFLAB GanedenBC30 viability on baking products during 2125
storage
C.-L. Jao, S.-L. Huang, S.-C. Wu, Hsu Kuo-Chiang
Formulation and characterization of biocompatible microemulsions as 2127
nutraceutics
A. Xenakis, V. Papadimitriou, T.G. Sotiroudis
Processing and technological characterization of extruded breakfast cereal 2129
obtained with a mix of broken rice and common bean flour
A.V. Carvalho, P.Z. Bassinello, A. de O. Rios
Cereal bar development using exotic fruit 2131
E. Rabelo Torres, E. Santana Castro, R. Felix de Santana, J. Cordeiro Cardoso, C.M.
Faria Soares, Á.S. Lima
Substitution of ingredients by green coconut (Cocos nucifera L) pulp in ice 2133
cream formulation
A.M. Iguti, A.C.I. Pereira, L. Fabiano, R.A. F. Silva, E.P. Ribiero
Evaluation of drying green coconut pulp for obtaining a snack-like product 2135
W.H. Prieto, E.A.G. Seravalli, A.M. Iguti, M. Nitz
Physical-chemistry and microbiological analysis of probiotic dairy beverage 2137
fermented with kefir
L.R. Ito Morioka, M. de Fátima Fonseca, L. Avallone Bueno, D. Marques, G. Cruz
Ximenes, C. Souza, M. Antônio de Morais Jr
Phytochemicals and antioxidant activity of comminuted orange (Citrus 2139
sinensis L.)
Z. Escobedo-Avellaneda, V. Serment-Moreno, A. Valdez-Fragoso, H. Mujica-Paz, J.
Welti-Chanes
Possibility of using durum wheat flour as an improvement agent in bread 2141
making process
A. Torbica, M. HadnaĀev, T. Dapÿeviý HadnaĀev
Sensory and antioxidant properties of beer with Juniperus communis L. 2143
M. Veljovic, S. Despotovic, R. Djordjevic, S. Pecic, A. Kalusevic, I. Leskosek-
Cukalovic, V. Nedovic

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxxiii
Influence of phytosterols addition in the rheology and sensory attributes of 2145
dark chocolate
P. Efraim, G.C. Marson, D.C.P. Jardim, A.O. Garcia, K. Yotsuynagi
Addressing new functional fillo products through nutrition and healthy 2147
ingredients: Hi omega-3 fatty acids and phytosterol esters
T. Varzakas, A. Labropoulos, S. Anestis
The non–starch polysaccharides quantity changes in pastry products where 2149
Jerusalem artichoke (Helianthus tuberosus L.) added
I. Gedrovica, D. Karklina, A. Fras, O. Jablonka, D. Boros
Characterization of cookies formulated with rice and black bean extruded 2151
flours
P.Z. Bassinello, D.De G.C. Freitas, J.L.R. Ascheri, C.Y. Takeiti, R.N. Carvalho, S.N.
Koakuzu, A.V. Carvalho
Isolation of lactic acid bacteria in Marajoara cheese, Amazon, Brazil 2153
H. Mendes de Figueiredo, C. Gonçalves e Gonçalves, P.C. de Moura Guimarães, A.
Mendes de Figueiredo Jr
The physico-chemical and microbiological aspects in ice-cream of buffalo 2155
milk added for fiber food
G.C. B. Chinelate, D.F. Pontes, R.R. de A. Bezerra
Process optimisation of egg replacer in sponge cake baking 2157
L. Mai, T. Norton, W. Li, B. Tiwari, C. Brennan
Obtaining functional fermented beverages by using the kefir grains 2159
T. Balabanova, P. Panayotov
Effect of synthesis conditions of short-chain fructooligosaccharides to obtain 2161
high yield and volumetric productivity
R. Vega, M.E. Zúniga-Hansen
Effect of pH culture on growth and fatty acid profile of Lactobacillus 2163
plantarum bacteria
C. Soto

Engineering of delivery systems of bioactive foods
Quality decay and viability of Lactobacillus acidophilus free and 2165
encapsulated in buffalo milk yogurt
A.S. Shoji, A.C. Oliveira, M.A. Trindade, O. Freitas, M. Thomazini, R.J.B.
Heinemann, C.S. Favaro-Trindade
Supercritical fluid extraction with modifier of antioxidant compounds from 2167
jabuticaba (Myrciaria cauliflora) by-product: economic viability
R.N. Cavalcanti, P.C. Veggi, M.A.A. Meireles
Microencapsulation of sacha inchi (Plukenetia volubilis L.) oil with zein 2169
S. Quispe-Condori, M.D.A. Saldaña
Encapsulation of curcumin loaded oil droplets by cryotropic gel formation 2171
from o/w emulsion
K. Nakagawa, N. Sowasod, T. Charinpanitkul, A. Soottitantawat, W.
Tanthapanichakoon

xxxiv
Effect of different ratios of maltodextrin/gelatin and ultrasound in the 2173
microencapsulation efficiency of turmeric oleoresin
C.R. Malacrida, V.R. Nicoletti Telis
Encapsulation of Melissa Officinalis leaf’s active compounds in Ƣ- 2175
cyclodextrin and modified starch
I. Mourtzinos, S.E. Papadakis, P. Igoumenidis, V.T. Karathanos
Deployment of response surface methodology to optimize recovery of 2177
grape (Vitis vinifera) stem and seed polyphenols
E. Karvela, D.P. Makris, N. Kalogeropoulos, V.T. Karathanos
Production of 1-octen-3-ol by Neurospora species isolated from beiju in 2179
different culture medium
D.S. de Carvalho, A.P. Dionísio, R. dos Santos, S. Boguzs Jr, H.T. Godoy , G.M.
Pastore

Food Product Engineering and Functional Foods

Characterisation of a non-alcoholic beverage made of residues from king 2181
palm (Archontophoenix alexandrae) industry
K. Cardoso Tramonte, J.G. Provesi, I. Moreira Dutra Albuquerque E Silva, A.
Nairne Negrão Murakami, M. Maraschin, R. Dias De Mello Castanho Amboni,
E.R. Amante
Composition of aroma compounds in fermented apple juice: effect of apple 2183
variety, fermentation temperature and inoculated yeast concentration
R. Riekstina-Dolge, Z. Kruma, D. Karklina, D. Seglina
Mode of inhibition of ơ-glucosidase and ơ-amylase by polyphenol-enriched 2185
extracts of maqui (Aristotelia chilensis)
F. Acevedo, M. Rubilar, B. Palma, C. Shene
Influence of pH variation during propolis extraction with the use of water as 2187
solvent
B.C.B.S.Mello, P.M. Kakuda, M.D. Hubinger
Modifier effects on supercritical fluid extraction (SFE) of some Brazilian 2189
plants: Antioxidant activity and economical evaluation
P.C. Veggi, R.N. Cavalcanti, M.A.A. Meireles
Anthocyanin extraction from jabuticaba (Myrciaria cauliflora) skins by 2191
different techniques: Economical evaluation
P.C. Veggi, D.T. Santos, M.A.A. Meireles
Study of cleaning efficiency of organic microfiltration membranes by 2193
attenuated total reflectance infrared microspectroscopy
T.K. Gelaw, A. Trentin, C. Güell, M. Ferrando, S. de Lamo-Castellví
Comparative study on quality evaluation of buffalo meat slices incorporated 2195
with finger millet, oats and chickpea
M. Siddiqui, M.A. Khan
Microencapsulation of tocopherols in lipid matrix by spray chilling method 2197
O.Diaz Gamboa, A. Lireny Guaraldo Gonçalves, R.C. Grosso

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxxv
Amino acid profile of Sous vide cooked poultry breast meat products 2199
K. Ramane, R. Galoburda, V. Kreicbergs, I. Vanaga
Antioxidant activity and porphyran content in hydrothermal extracts of 2201
Porphyra Yezoensis (Susabinori)
C. Goto, S. Machmudah, M. Sasaki, M. Goto, K. Okai, Y. Okai, S. Kondo
Effect of frozen storage on the quality of camu camu (Myrciaria dubia (H. 2203
B.K.) McVaugh,) pulp
A.L.R. Souza, M.M Pagani, F.S. Gomes, L.M.C. Cabral
Effect of semolina particle size on the cooking kinetics and quality of 2205
spaghetti
G. Sacchetti, G. Cocco, D. Cocco, L. Neri, D. Mastrocola
Kinetics of heterogeneous amylolysis in oat flour and characterization of 2207
hydrolyzates
A. Patsioura, V. Gekas, A. Lazaridou, C. Biliaderis
Kinetics of Amycolatopsis mediterranei DSM 43304 lipase-mediated 2209
synthesis of isoamyl acetate in n-hexane
D.S. Dheeman, J.M. Frías, G.T.M. Henehan
PROBIOLIVES: Table olive fermentation with selected strains of probiotic 2211
lactic acid bacteria. Towards a new functional food (FP7-SME-2008- 2
project)
C.C. Tassou, E.Z. Panagou, A. Garrido-Fernandez, C. Peres, L. Cocolin, N.
Chammem
Effect of vacuum drying on blackcurrant’s antioxidant components 2213
M. Stéger-Máté, B. Nótin, R. Juhász, B. Verasztó, D. Jakab, J. Monspart-Sényi, J.
Barta
Production of bioactive metabolites with pharmaceutical and nutraceutical 2215
interest by submerged fermentation of pleurotus ostreatus in a batch stirred
tank bioreactor
L.-M. Papaspyridi, N. Aligiannis, P. Christakopoulos, A.-L. Skaltsounis, N.
Fokialakis

NOVEL FOOD PROCCESSES

Effects of High Intensity Pulsed Electric Fields or Thermal Treatments on 2217
Carotenoid Profile of a Fruit Juice-Soymilk Beverage along Chilled Storage
Laura Salvia-Trujillo, Mariana Morales-de la Peña, Ma. Alejandra Rojas-Graü, Olga
Martín-Belloso

Amino Acid Composition of a Fruit Juice-Soymilk Beverage as Affected by 2219

High Intensity Pulsed Electric Fields or Thermal Treatments during
Storage
Mariana Morales-de la Peña, Laura Salvia-Trujillo, Teresa Garde-Cerdán, Ma.
Alejandra Rojas-Graü, Olga Martín-Belloso

xxxvi
 HIGHTECH EUROPE PROJECT WORKSHOP
OPEN INNOVATION IN FOOD PROCESSING

Challenges and essentials for reinventing R&D in an open innovation 2221

ecosystem
I.S. Saguy
Philosophy of an open R&D system 2223
K. Chida
Creating value for SMEs in the food industry through open innovation - 2223
Examples from Norway
Ø. Fylling-Jensen
Open Innovation at Mars: Join-up, speed-up, scale-up 2223
O. Fleurot
Innovation sharing by cooperative R&D 2224
D. Albers
HighTech Europe Interactive Technology Portal – New tool for innovation in 2225
food processing
K. Lienemann, N. Ay, R. Groeneveld, D. Willems, I. Van der Plancken

INSIDEFOOD PROJECT WORKSHOP

NOVEL TECHNOLOGIES TO EXPLORE FOOD
MICROSTRUCTURE

Food microstructure: a 3-D experience 2227

B. Nicolaï
Possibilities of X-ray nano-CT for internal quality assessment of food products 2227
E. Herremans, S. Chassagne-Berces, H. Chanvrier, A. Atoniuk, R. Kusztal, E.
Bongaers, B.E. Verlinden, E. Jakubczyk, P. Estrade, P. Verboven, B. Nicolaï
Optical coherence tomography (OCT) for quality control and microstructure 2229
analysis in food
M. Leitner, G. Hannesschläger, A. Saghy, A. Nemeth, S. Chassagne-Berces, H.
Chanvrier, E. Herremans, B.E. Verlinden
Effect of fibres and whole grain content on quality attributes of extruded 2231
cereals
S. Chassagne-Berces, M. Leitner, A. Melado, P. Barreiro, E. Crostina Correa, I. Blank,
J.-C. Gumy, H. Chanvrier
NMR microscopy and NMR HR-MAS on apples of different qualities after 2233
different storage conditions
D. Gross, M. Spraul, E. Humpfer, H. Schaefer, A. Melado, T. Defraeye, P. Verboven

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxxvii
A Digital Laboratory for visual analysis of materials microstructure 2233
P. Estrade
Cryo scanning electron microscopy: enabling nano-imaging of food products 2233
F. Depypere, D. Van de Walle, K. Dewettinck
The application of acoustic emission to measure texture of food foams 2235
E. Jakubczyk, E. Gondek
Non destructive detection of brown heart in ‘Braeburn’ apples by time- 2237
resolved reflectance spectroscopy (TRS)
M. Vanoli, A. Rizzolo, M. Grassi, A. Farina, A. Pifferi, L. Spinelli, B.E. Verlindend,
A. Torricelli
Non-destructive characterization of food microstructure and composition by 2239
spatially resolved spectroscopy (SRS)
N. Nguyen Do Trong, M. Tsuta, E. Herremans, R. Watté, C. Erkinbaev, E.
Verhoelst, P. Verboven, B. M. Nicolaï, W. Saeys

FRISBEE PROJECT WORKSHOP

REFRIGERATION INNOVATIONS AND COLD CHAIN
MANAGEMENT

New tools, concepts and solutions for improving technologies along the 2241
European food cold chain: the FRISBEE project
G. Alvarez, A. Geeraerd, D. Leducq, J.Evans, E. Wissink, E. Indergård, C. Cotillon,
P. Taoukis
Management and optimization of the cold chain and the development of cold 2243
chain data base
P. Taoukis, G. Katsaros, T. Tsironi, E. Dermesonluoglu, E. Gogou
Towards a framework for evaluation of energy consumption, sustainability 2245
and associated food quality in the European cold chain
S.G. Gwanpua, B. Verlinden, S. van der Sluis, E. Wessink, J. Evans, T. Brown, D.
Leducq, G. Alvarez, P. Taoukis, G. Katsaros, V. Stahl, D. Thuault, I. Claussen, E.
Indergård, P. Verboven, B. Nicolaï, A. Geeraerd
Influence of room temperature on food safety in refrigerated display cabinet 2247
O. Laguerre, M. Hoang, G. Alvarez, D. Flick
Improvement of existing concepts and refrigeration technologies: advanced 2249
control and thermal energy storage applied to food refrigeration
D. Leducq, P. Shalbart, F. Trinquet, A. Graciela, B. Verlinden, S. van der Sluis, E.
Wessink, J. Evans, T. Brown, B. Nicolaï, A. Geeraerd, P. Verboven, J. M Lagaron, F.
Jay, M. Pirani, E. Indergård
Emerging refrigeration technologies at laboratory scale to improve food 2251
quality and reduce environmental impact and energy consumption
J. Evans, T. Brown, D. Leducq, G. Alvarez, P. Verboven, B. Nicolaï, A. Geeraerd, E.
Wessink, I. Claussen, E. Indergård, J.M. Lagarón, R. Pérez Masiá, S. Mousset, A.
Soysal, M.-C. Zelem, N. Wilson

xxxviii
The potential for superchilling to enable safe, high quality and long term 2253
storage of foods
I.C. Claussen
CAFÉ : Computer-Aided Food processes for control Engineering, European 2255
project CAFÉ
D. Dochain, A. Antonio

Design and development of REAlistic food Models with well-characterised 2257

micro- and macro-structure and composition: European project DREAM
M. Axelos

PROSPARE PROJECT WORKSHOP

INNOVATIVE FUNCTIONAL PROTEINS FROM
POULTRY LEFTOVERS

Innovative technologies from Animal-by Products bioconversion European 2259

project PROSPARE
A. Dossena, V. Popov
The Animal by Product (AB-P): challenging problem and resource 2263
W. De Roover
Approach and objectives of the PROSPARE project 2263
O. Koroleva
Innovative methodology and process technologies 2263
O. Koroleva
Molecular composition and functional properties of poultry hydrolyzates 2263
obtained in the PROSPARE Project
A. Dossena
Food & feed market exploitation and nutrition 2263
V. Guardiani

Authors Index

International Association for Engineering and Food

List of country delegates

11th International Congress on Engineering and Food - Athens, Greece, 2011 xxxix
xl
 Mass Transfer of Fruit Slices in Hypertonic Solution

Farid Amidi Fazlia, Neda Amidi Fazlib

a
Food Science and Technology Department of Islamic Azad University, Soofian Branch, Iran
(amidi_f@yahoo.com)
b
B.Sc student of Food Science and Technology, Tabriz, Iran

INTRODUCTION
Emerging of fruits in concentrated solutions cause counter current flow between fruit and
syrup, the water flow from food material to surrounding syrup and solutes from syrup to food.
The result of this process is semi dehydrated food with higher dry matter content in compare
with untreated food; the other aspect of this process is self formulation of food staff or
enrichment of it by adding especial and desirable osmosis agents like antioxidants or minerals.
The healthy foods are demanded by consumers in recent years; enriched fruits by minerals are
a kind of these products. They provide minerals which are needed to human body; the minerals
are used by body cells for daily activities as well as cell maintenance, minerals have important
role in precise periods of life like growth and mature ages. By osmosis process it is possible to
obtain natural enriched food materials with any type of minerals in desirable content.
The effect of calcium lactate on osmotic dehydration kinetics and on the respiration rate,
mechanical properties and shelf-life of fresh, vacuum impregnated (VI) and pulsed vacuum
osmodehydrated (PVOD) grapefruit has studied by moraga et al. [1]. Apple cylinders were
vacuum impregnated, osmodehydrated and rehydrated using solutions of glucose, sucrose and
trehalose. The kind of solute affected mass transfer rate and the glucose solution showed the
lowest kinetics [2].

MATERIALS & METHODS

This paper indicates the result of osmotic dehydration of kiwi fruit and banana slices in 55%
sucrose syrup in presence of calcium and phosphorus ions. Osmosis agent (55% (w/w)) was
prepared by solving sucrose in deionized water and adding calcium or phosphorus in 1 and 2%
concentration. 1 centimeter in thickness sliced fruits emerged in above syrup and process
completed in 30 and 60 minutes. Dry matter was determinate for untreated and treated samples
using 100 ÛC oven. Osmosis parameters as solid gain (SG), water loss (WL) and weight
reduction (WR) were calculated using below equations [3], where W0, W, S0 and S are initial
and final sample weight and initial and final sample dry matter respectively.
WR= (W0-W)/W0
SG= (S-S0)/W0
WL= WR+SG

RESULTS & DISCUSSION

Results (table 1) showed that highest mass transfer in kiwi fruit occurs when 1% calcium
solution applied for 60 minutes, values obtained for solid gain, water loss and weight reduction
were 42.60, 51.97 and 9.37 respectively in kiwi fruit where highest values for solid gain and
water loss were 35.25 and 41.07 respectively.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1609
 Table 1. mass transfer results for kiwi fruit and banana in different osmotic solutions
Concentration Time SG (%) WL (%) WR (%)
Mineral
(%) (min) Kiwi fruit Banana Kiwi fruit Banana Kiwi fruit Banana
Ca 1 30 22.83 3.541 29.53 7.636 6.701 4.095
60 42.6 8.729 51.97 13.25 9.369 4.523
2 30 30.25 2.5 37.71 6.498 7.464 3.998
60 27.51 6.584 36.44 11.31 8.927 4.725
P 1 30 35.25 12.62 41.07 15.86 5.822 3.24
60 33.14 10.75 40.35 15.47 7.207 4.724
2 30 25.25 8.509 30.99 12.79 5.737 4.28
60 23.36 21 30.59 25.84 7.23 4.835

when 1% phosphorus applied for 30 minutes, highest weight reduction in kiwi fruit was 7.23
when 2% phosphorus for 60 minutes was used as osmosis agent. Higher mass transfer in the
case of calcium may be due to small ion size of calcium. Adding calcium to solution can
provide higher molarity to phosphorus and consequently higher osmosis pressure can be
reached in solutions, higher osmosis pressure can lead to increased mass transfer as seen in
kiwi fruit dehydration by osmosis treatment using different minerals as osmosis agent.
Results of osmosis treatment of banana slices have showed in table 1. When 2% phosphorus
concentration was applied as osmosis agent for 60 minutes highest values for solid gain, water
loss and weight reduction obtained as 21, 25.84 and 4.84 respectively. When calcium was used
as osmosis agent mass transfer parameters were lower than mass transfer parameters when
phosphorus was used as osmosis agent. Solid gain, water loss and weight reduction reached to
8.73, 13.25 and 4.72 respectively when calcium was osmosis agent. Regarding table 1 and 2
can be lead to this fact that osmosis parameters in banana were lower than kiwi fruit osmosis
parameters in both calcium and phosphorus as applied as osmotic dehydration agent, this may
be due to different textural properties of mentioned fruits.

CONCLUSION
It seems this process is an appropriate way to produce enriched fruits by minerals and ensure
that body obtains enough and required quantity of minerals. Further research are recommended
on different minerals and any other micro nutrients; also in vivo studies are necessary to
determinate intake amount of such nutrients. More studies need to investigate different fruit
physicochemical properties of fruits which affect mass transfer during osmotic dehydration
process in plant tissues.

REFERENCES
[1] M.J. Moraga, G. Moraga, P.J. Fito, N. Martínez-Navarrete, 2009. Effect of vacuum impregnation with
calcium lactate on the osmotic dehydration kinetics and quality of osmodehydrated grapefruit. Journal of
Food Engineering, 90, 372–379. [2] L. Atarés, A. Chiralt, C. González-Martínez, 2008. Effect of solute
on osmotic dehydration and rehydration of vacuum impregnated apple cylinders (cv. Granny Smith).
Journal of Food Engineering, 89, 49–56. [3] Mavroudis, N. M., Gekas, V. Sjoholm, I. 1998. Osmotic
dehydration of apples: effect of agitation and row material characteristics. Journal of Food Engineering,
35, 191-209.

1610
 Nanofiltration treatment of waste brine obtained from sugar decolorizing resin

regeneration

Fakhreddin Salehi, Seyed M.A. Razavi

Department of Food Science and Technology, Ferdowsi University of Mashhad,

Khorasan, Mashhad, Iran (FS1446@Yahoo.com, S.Razavi@um.ac.ir)

INTRODUCTION
In the sugar industry, anion exchange resins are mainly used to remove high molecular weight
colorants like melanins, melanoidines, caramels and polyphenols with typical molar mass of
colorant ranges from 500 to 20000 D from cane sugar liquor. The colorant are at first adsorbed
onto the resins, and finally resins is regenerated by passing an alkaline brine solution [1, 2].
The waste brine stream arising from the regeneration of the resins usually imposes a serious
problem of disposal and significant environmental costs since the effluent contains a high
amounts of sodium chloride (50-100g/1), organic matter (5g/l as total carbon) and chemical
oxygen demand (COD, about 13000 mg/l) [1-3]. To counter these problems, nanofiltration
process has been successfully applied to recycle spent caustic CIP cleaning fluids and spent
anion exchange [4]. The aim of this paper was to study the performance of polyamide NF
membrane for the purifying of waste brine from resin regeneration to (i) investigate the
influence of feed concentration in a wide range as well as operating conditions (temperature
and transmembrane pressure) on permeate flux and rejections of NaCl and dyes, (ii) recover
brine for reuse, reduction in water consumption and effluent volume.

MATERIALS & METHODS

The polymeric tubular AFC80 membrane was supplied by PCI membrane systems, USA, made
from of polyamide film, was used in this study. The operating pressure of each run was at the
range of 1.0–2.0 MPa (at five levels). The temperature was varied from 30°C-50°C (at three
levels) and controlled by a tubular heat exchanger. In order to investigate the effect of feed
concentration on the permeate flux and rejection; feeds were prepared at four concentration
levels (40, 60, 80 and 100 g/l). Optical density (OD) was chosen as a measure for the colorant
concentration in the permeate and retentate samples [1]. The OD was measured at 420 nm and
25oC using a UV-VIS spectrophotometer (Jenway 6105, Bibby Scientific Limited, UK).

RESULTS & DISCUSSION

Result showed the permeate rate increases with increasing pressure and temperature and
decreases with increasing feed concentration. Hong et al (2006) [5] found the solution flux
decline with increasing NaCl concentration because of an increasing osmotic pressure drop
across the membrane. The sodium chloride rejection was decreased by increasing feed
concentration and temperature, whereas it increased with an increase in transmembrane
pressure. Fig. 1 shows the effect of temperature at different feed concentrations on the salt
rejection. The results show that the rejection of NaCl was greatly decreased from 37% to 19%
with increase in feed concentration from 40 to 100 g/l at 1.5MPa and 30°C. In the present
work, the highest rejection of salt for nanofiltration processing of waste brine was determined

11th International Congress on Engineering and Food - Athens, Greece, 2011 1611
as 37% at 40g/l, 2.0MPa TMP and 30°C. The polyamide NF membrane had not a serious
problem of fouling and the flux was approximately constant during whole membrane process
of waste brine. Dye rejection values were obtained by measuring the dye concentration in the
samples by UV-Visible spectroscopy. Uniformly, complete colorant removal (>99.9%) was
achieved in this study, whereas about 77% of the salt was recovered at 1.0MPa TMP, 50°C,
100g/l and the feed was concentrated up to VCF9.
40
35
NaCl rejection (%)

30
25
20
15
10
5 40g/l 60g/l 80g/l 100g/l
0
20 30 40 50 60
Temperature (°C)

Figure 1. Effect of feed concentration and temperature of nanofiltration process on NaCl rejection of
regeneration waste brine (ǻP=1.5MPa)

CONCLUSION
The trials display satisfactory qualitative performance for the removal of colorant from the
resin regeneration waste and recovery of salts for reuse. The NF membranes allowed the
achievement of a 77% reduction in salt consumption and an 90% reduction in water
consumption and removed of >99% of colorant components. Totally, the permeate flux
increased almost 0.538 kg/m2.h and 0.204 kg/m2.h as the pressure and temperature increased
0.1MPa and 1oC, respectively, however it decreased approximately 0.456 kg/m2.h when the
feed concentration increased by 10g/l. On the other hand, the rejection of salt by polyamide NF
membrane was declined about by 0.505% and 1.42% with 1oC and 10g/l increase in
temperature and concentration, respectively, whereas it increased averagely 0.65% as the
transmembrane pressure increased as 0.1MPa. The stable flux, high rejection of colorant and
high NaCl recovery show that polyamide NF membrane is a very promising method of
treatment for this type of waste brine.

REFERENCES
[1] Cartier S., Theoleyre M.A. & Decloux M. 1997.Treatment of sugar decolorizing regeneration waste using
nanofiltration. Desalination, 113, 7–17.
[2] Hinkova A., BubnÕk Z., Kadlec P. & Pridal J. 2002. Potentials of separation membranes in the sugar industry. Sep.
Purif. Technol., 26, 101–110.
[3] Wadley S., Brouckaert C.J., Baddock L.A.D. & Buckley C.A. 1995.Modelling of nanofiltration applied to the
recovery of salt from waste brine at a sugar decolorisation plant. J. Membr. Sci., 102, 163–175.
[4] Durham R.J., Sleigh R.W. & Hourigan J.A. 2003. Nanofiltration for recovery of spent ion exchange brines,
IMSTEC’03, 5th International membrane science and technology conference, Sydney.
[5] Hong S., Miller M.D. & Bruening M.L. 2006. Removal of Dyes, Sugars, and Amino Acids from NaCl Solutions
Using Multilayer Polyelectrolyte Nanofiltration Membranes. Ind. Eng. Chem. Res., 45, 6284-6288.

1612
 Process Development of Ready-to-eat Custard Cream Filled Chinese Steamed Bun

Chaiwanichsiri, S.a, Poonnakasem, N.a,b, and Laohasongkram, K.a,c

a
Department of Food Technology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand
(saiwarun.c @chula.ac.th)
b
(pomac116@hotmail.com)
c
(kalaya.l@chula.ac.th)

INTRODUCTION
Chinese steamed bun is a popular food in Asia. However, its storage life is short as it has high
moisture content. Freezing is sometimes used to prolong its shelf life but the energy
consumption is high. Hurdle technology is a combined preservation method by applying
different means such as adjusting water activity (aw), pH, temperature, redox potential,
modified atmosphere. Therefore, this study aimed to develop Chinese steamed bun with
custard cream filled (CCSB) that can be kept at room temperature for about 10 days by using
hurdle technique.

MATERIALS & METHODS

The hurdle parameters used were aw, pH, and modified atmosphere packaging (MAP).
Glycerol and fructose were added into the custard cream filling at 3-9% of total weight and aw,
and sensory qualities were evaluated. A 3x3 factorial design experiment was used and the
optimum conditions of the additives were determined from the response surface methodology
(RSM) on sweetness, texture, and overall acceptance (OAA). For the steamed bun, glycerol
(0-5%) and lactic acid (0-5%) were added to the dough during a two-step dough preparation.
The aw, pH, and sensory qualities (appearance, flavour, texture and overall acceptability) of the
buns were monitored. The effect of preservative, calcium propionate (CaP), on aw, pH, and
microorganism (aerobic bacteria, yeast and mold) was studied in the bun. Lastly, the shelf-life
of the custard cream Chinese steamed bun (CCSB) packaged with and without oxygen
absorber (OA) was investigated at room temperature (30 + 2oC). The aerobic plate count (AC),
aw, pH, texture and sensory quality were determined every 2 days until the CCSB spoiled or
was not accepted.

RESULTS & DISCUSSION

It was found that water activity (aw) of the custard cream decreased with increasing glycerol
and fructose while the sensory scores increased with increasing glycerol and fructose up to 6%
and then decreased after that. From RSM it was found that the optimum condition for custard
cream filling was the addition of 6% glycerol and fructose each, which could reduce aw to
0.915. For the Chinese steamed bun, the analysis of variance (ANOVA) showed that there was
no interaction effect of glycerol and lactic acid so each effect was analyzed separately. When
we consider the effect of glycerol it was found that aw of the bun decreased significantly as
glycerol increased from 0 to 5% (p < 0.05) presumably due to the lowering free water in the
dough. The pH and all sensory scores except appearance were not affected by the glycerol and
lactic acid (p > 0.05). When we consider the effect of lactic acid it was found that pH, flavour

11th International Congress on Engineering and Food - Athens, Greece, 2011 1613
and OAA of the bun decreased with increasing lactic acid (p<0.05) while aw, appearance, and
texture did not change significantly (p>0.05). Since the bun with 0.25% lactic acid had
sensory score on flavour and OAA insignificantly different from control (0% glycerol and
lactic acid). Therefore, buns having 2.5% glycerol and 0.25% lactic acid were prepared with
calcium propionate (CaP). The result showed that CaP did not affect the properties and shelf-
life of the bun. The aerobic plate count of the bun after storage for 16 days was less than 4 log
CFU/g which was lower than the acceptable limit of 5 log CFU/g. Therefore, the custard
cream having 6% glycerol and 6% fructose was filled into the Chinese steamed bun which had
2.5% glycerol and 0.25% lactic acid. The CCSB was individually packaged in a PVDC bag
with and without oxygen absorber (OA). The results showed that OA significantly slowed
down the growth of aerobic microorganism and there was no yeast and mold detected. The
CCSB packaged with OA could extend up to 10 days. However, from the textural
measurement the acceptable level of the sample was only 8 days. Thus the shelf-life of the
hurdle-treated CCSB without CaP packed with OA was 8 days.

CONCLUSION
The CCSB process was developed to produce the shelf-stable product using hurdle technology.
The optimum levels for custard cream filling were 6% of glycerol and fructose each. Addition
of 2.5% glycerol and 0.25% lactic acid reduced aw and pH of Chinese steamed bun to 0.912
and 5.78, respectively, and the sensory qualities were not significantly different from the
control. Based on microbiological test, this hurdle-treated bun could be stored for 16 days at
room temperature without preservative. The hurdle-treated CCSB with oxygen absorber in
PVDC packaging could be stored for at least 10 days at room temperature without microbial
spoilage. But the texture of the product was unacceptable. From the microbiological and
textural viewpoints, the shelf-life of the developed CCSB was extended to at least 8 days at
room temperature (30 ± 2OC).

REFERENCES
[1] Peng Q., Shun-he C., & Chuan M. 2007. Effect of Waxy Wheat Four Blends on the Quality of
Chinese Steamed Bread. Agricultural Sciences in China, 6, 1275-1282.
[2] Leistner L. & Gorris L.G.M. 1995. Food Preservation by Hurdle Technology. Trend in Food
Science and Technology, 6, 41-46.
[3] Das H. & Radhakrishna K. 2001. Preservation of Mutton as Ready-to Eat Curry by Hurdle
Technology. Journal of Food Science and Technology, 38, 287–289.
[4] Lombard G. E., Weinert I. A. G., Minnaar A., & Taylor J. R. N. 2000. Preservation of South African
Steamed Bread Using Hurdle Technology. Lebensmittel Wissenschaft Und Technologie Food
Science and Technology, 33, 138-143.

1614
 Decontamination of spices by using a pulsed light treatment

M. Moreaua, I. Nicorescua, A. S. Turpina, A. Agoulonb, S. Chevaliera, N. Orangea

a
Laboratoire de Microbiologie du Froid-Signaux et Micro-environnement EA 4123, 55 rue Saint
Germain, 27000, Evreux, France (irina.nicorescu@univ-rouen.fr)
b
AgroHall, 55 rue Saint Germain, 27000, Evreux, France (adrien.agoulon@agrohall.fr)

INTRODUCTION
For pasteurising dried food products, the most often employed decontamination techniques are
high temperature short time treatments, UV light irradiation, dry heat, steam, microwave and
IR heating. However, microbial reduction by using pulsed light (PL) is gaining researcher’s
attention because of the request of less of energy compared to thermal processes. PL
technology relies on a series of very short, high-power pulses of broad-spectrum light, typically
emitted by xenon lamps, to destroy bacteria (both vegetative cells and spores), yeasts, moulds
and even viruses. Pulsed light treatment is effective for the inactivation of bacteria (vegetative
cells and spores) on surfaces, packaging and recently in food products (meat, bread, vegetables,
and fruits). Indeed, Ozer and Demirci (2006) have reported a 1.09 log reduction for Listeria
monocytogenes on raw salmon filets after 180 pulses of light and, Sauer and Moraru (2009)
achieved a 7.15 log CFU reduction of Escherichia coli in apple juice. However, in the
literature, there is still a lack of knowledge concerning the decontamination of
powdered/granular foods by using pulsed light. Consequently, the main aim of our work was to
evaluate the effectiveness of a PL treatment on the inactivation of B. subtilis in spices.

MATERIALS & METHODS

Strain and cultivation conditions
B. subtilis spores supplied by INRA Dijon were dispersed in trypton salt medium (TS) in order
to obtain a 109 spores/mL concentration. Glass marbles (10g) and black peppercorn (25g) were
inoculated with spore suspension containing 108 spores/mL of B. subtilis, dried by using a hair
dryer and then, pulsed light treatment was carried out. The control and flashed samples (2, 4, 6,
8 and 10 flashes of light) were resuspended in TS medium and shaked during 15 min. to
reconstitute the cellular suspension. Decimal dilutions were spread on Colombia-agar medium.
Viability was evaluated by counting bacteria colonies (CFUs) and comparing with control
samples, which were prepared in the same conditions but not exposed to light flashes.

Pulsed light treatment

The PL experimental set-up (Claranor, France) is composed of a power supply unit and a flash
lamp. The treatment chamber contains four cylindrical xenon lamps. Samples were treated at
3000V and with 2, 4, 6, 8 or 10 pulses of light. For each pulse the duration was of 300 Ps and
the energy level measured by using a calorimeter Solo-2 (Gentec-EO, Canada) was found to be
of 1.06 J.cm-2. After exposure to PL, treated and untreated samples were analysed immediately.

Measurement of temperature profile

Control and flashed samples were taken in picture by using an IR camera FLIR T360 (FLIR
Systems France, France) in order to determine the radiation power and consequently, the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1615
temperature profile when applying a pulsed light treatment in our experimental conditions.
Images were then analysed by using a ThermaCAM Reporter software.

RESULTS & DISCUSSION

Temperature profile
Experimental results obtained on the glass marbles allowed us to verify the hypothesis that the
decontamination effect produced by pulsed light treatment was not given by the increase in
temperature but only by the action of UV rays. Indeed, this study allowed us to notice that
glass marbles were not heated during the PL treatment, since no significant temperature rise
was registered after 10 pulses of light at 3000 V. However, when an identical PL treatment was
applied to black peppercorn samples, a significant increase (13.5 °C) in the temperature profile
was observed, especially for a 10 pulses of light treatment. Concerning the 6 pulses of light
treatment applied on black peppercorn sample, the temperature rise recorded was of around 9
°C, these experimental data suggest that in the future, it will be wiser to choose this last
treatment and thus limiting the impact of the temperature rise on the food matrix.

Impact of Pulsed light treatment

Concerning the decontamination of glass marbles by pulsed light, experimental data found can
be interpreted as follows:
x 2 pulses of light leads to a decimal reduction of about 0.9 – 1.1 log CFU
x a reduction between 1.7 and 2.1 log CFU was obtained for a treatment of 4 pulses
x for a treatment superior to 6 pulses of light, decontamination is almost identical and
stabilizes around 2.6 log CFU for the glass marbles of 2 mm in diameter and around
2.8 log CFU for those of 4 mm in diameter.
Pulsed light treatment effects were greater in glass marbles than peppercorn decontamination.
Indeed, the destruction level remained below 1 log even when a treatment of 10 pulses was
carried out on the peppercorn samples. However, a slight improvement (> 0.4 log CFU) in
spice microbial reduction with increasing pulse number indicated that this parameter could be
investigated.

CONCLUSION
Experimental results showed that it is possible to find conditions that lead to an optimal
decontamination in the case of glass marbles (2.7 log CFU). Similar results were found when
the pulsed light treatment was carried out on black peppercorn, only that the decimal reduction
achieved was less important comparing to glass marbles (0.8 log CFU). However, our results
suggest that PL technology should be also efficient onto naturally infected spices.

REFERENCES
[1] Ozer, N.P., & Demirci, A. 2006. Inactivation of Escherichia coli O157:H7 and Listeria
monocytogenes inoculated on raw salmon filets by pulsed UV-light treatment. International Journal
of Food Science and Technology, 41, 354-360.
[2] Sauer, A., & Moraru, C.I. 2009. Inactivation of E. coli ATCC 25922 and E. coli O157:H7 in apple
juice and apple cider, using pulsed light treatment. Journal of Food Protection, 72, 937-944.

1616
 Acceleration of precipitation formation in peach juice

induced by high-pressure carbon dioxide

Linyan Zhoua, Yan Zhanga, Xiaojun Liaora, Xiaosong Hua

a
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China
(Liaoxjun@hotmail.com)

INTRODUCTION
The destabilization of cloudy juices by High pressure carbon dioxide (HPCD) has been
known[1], which could reduce appearance quality of the juice. In order to better understand the
acceleration of the precipitation formation and the role of the HPCD, the focus of the present
work is to analyze factors including particle size distribution, pH and ȗ-potential, contributing
to the precipitation formation, and to compare the effects of HPCD and heat on the stability of
the peach juice.

MATERIALS & METHODS

Peaches (Cultivar No.24 Beijing) were purchased from Beijing Guangyuan Yanwei
Agricultural Science and Technology Co., Ltd. The halves of peaches were pitted and slices by
hand, juiced with a screw juice extractor (GT6G7, Zhejiang Light Industry Machinery Plant,
Zhejiang, China), and filtrated with 4 layers of cheese cloth.
A 400 mL sample of peach juice was treated by HPCD and heat treatment. The treatment
parameters applied in this study were as bellows: the pressure was 30 MPa, the temperature
was 55 ć, and the treatment time was 30 s, 10, 40, and 60 min. The time of precipitation
formation was estimated immediately after HPCD or heat. Parameters contributed to
precipitation formation was analysed after HPCD and heat.

RESULTS & DISCUSSION

3
Volume (%)

1 10 100 1000

Particle diameter O ­mP

Figure 1. The PSD patterns of peach juices treated by HPCD at 30 MPa and 55ć and heat at 90 ć for 1
min.
untreated; 30 s; 10 min; 40 min; 60min; heat treatment

11th International Congress on Engineering and Food - Athens, Greece, 2011 1617
Figure 1 compared the PSD patterns of peach juices after HPCD and heat. As shown in Figure
1, the PSD patterns were significantly changed after HPCD. All the HPCD-treated juices
showed the widest PSD patterns from 0.3 to 600 ȝm with three volume peaks, and the highest
volume peak was shifted to 18~21 ȝm. The particle size of the HPCD-treated juices
significantly increased as compared with the untreated and heat-treated juices. The volume
value of the HPCD-treated juices all showed a noticeable decrease for the smaller particles and
increase for the larger particles. With increasing the treatment time, the volume peak values
showed an increase for the smaller particles and a decrease for the larger particles, meanwhile
the particle sizes of the highest volume peak showed a left shift. Table 1 showed that d43 and
d32 of the HPCD-treated juices were both significantly larger than those of the untreated and
heat-treated juices, and exhibited a decrease with increasing the treatment time.

Table 1. Characteristics of peach juice treated by HPCD

precipitation d43 d32 pH ȗ-potential Ca
formation (ȝm) (ȝm) (mV) (mg/L)
time (h)
Untreated 60 ± 3 2.15 ± 0.01d 1.24 ± 0.01d 3.82 ± 0.02a -6.76 ± 0.36a 31.8
30 s 6±1 28.28 ± 0.45a 6.80 ± 0.14a 3.83 ± 0.01a -6.68 ± 0.25a 28.7
10 min 5±1 22.31 ± 0.51b 4.53 ± 0.08b 3.83 ± 0.02a -6.56 ± 0.34a 31.6
40 min 5±1 19.31 ± 1.35bc 3.41 ± 0.01c 3.78 ± 0.03a -6.29 ± 0.25a 34.65
60 min 7±1 17.65 ± 1.53c 3.19 ± 0.37c 3.80 ± 0.03a -6.92 ± 0.26a 31.8
Heat(90
60 ± 2 3.66 ± 0.24d 1.19 ± 0.03d 3.82 ± 0.02a -6.54 ± 0.20a 34.25
,1 min)
peach juices treated by HPCD at 30 MPa and 55 ć for 30 s, 10 min, 40 min, 60 min, and heat at 90 ć for 1 min.

In the present work, no effects of HPCD and heat could be found on pH and ȗ-potential of
peach juices (Table 1). The values of the ȗ-potential were negative, and kept constant around -
6.7 mV. It should be noted that the above ȗ-potential and pH were measured after
decompression. In fact, the pH of the system could be significantly reduced by the carbonic
acid from the dissolved CO2 during the HPCD processing[2]. Therefore, it was reasonable to
believe that the pH and the absolute value of ȗ-potential in the peach juice could be lower than
3.8 and 6.7 during HPCD, respectively, and these declines possibly induced the coagulation of
protein in the juices, responsible for the alteration of the PSD patterns and the acceleration of
the precipitation formation in the HPCD-treated juices.

CONCLUSION
These results suggested that pH and the absolute value of ȗ-potential decline induced the
coagulation of protein and decrease of particle charge, responsible for the acceleration of the
precipitation formation.

REFERENCES
[1] Zhou L. Y., Wang Y. Y., Hu X. S., Wu J. H., & Liao X. J. 2009. Effect of high pressure carbon
dioxide on the quality of carrot juice. Innov. Food Sci. Emerg. Technol. 10, 321-327.
[2] Calix T. F., Ferrentino G., & Balaban, M. O. 2008. Measurement of high-pressure carbon dioxide
solubility in orange juice, apple juice, and model liquid foods. J. Food. Sci. 73, E439-445.

1618
 Effect of the electric field on the vitamins A, C and E alone and added to avocado paste.

Raúl René Robles de la Torrea, María Guadalupe Méndez Ramosa, Ma. Reyna Robles Lópeza, José
Alberto Ariza Ortegaa, Francisco Javier Martínez Montesa.

a
Centro de Investigación en Biotecnología Aplicada-IPN. Ex–Hacienda San Juan Molino, Km. 1.5.
Carretera Tepetitla-Tecuexcomac, Tepetitla, Tlaxcala. C.P. 90700. México. (rrenerdlt@yahoo.com)

INTRODUCTION
For many decades, methods to preserve avocado pulp have been sought; any method must
inhibit the activity of the polyphenol-oxidase enzyme and at the same time retain the sensory
and nutritional characteristics of the fresh product. Thermal treatments are not recommended
because they reduce the excellent sensory characteristics and increase the nutritional losses of
the fruit. The use of chemical preservatives has also been banned by several international
regulations. Due to that, the use of emerging technologies have been proposed, these
technologies do not use thermal energy or use it more efficiently, they are usually applied for
short times, and, it has been observed, they can preserve the fresh product characteristics.
Among the emerging technologies are the high hydrostatic pressure, the pulsed electromagnetic
fields, the ultrasound and more recently the electric fields, (EF). In a previous work it was
demonstrated the effective use of the electric field to inhibit the polyphenol-oxidase (PPO)
enzyme in avocado pulp. Therefore the objective of this work was to study the effect of the
electric field on standards of E-carotene, D-tocopherol and ascorbic acid, and on these same
vitamins added to avocado paste as internal standards.

MATERIALS & METHODS

In this study a generator of electric field device was used, it was designed and built by the
group of the CIBA-IPN, a unit of the National Polytechnic Institute, located in Tlaxcala,
México. This equipment has the four key elements to generate the EF: a source of high voltage,
a frequency modulator, a solid state relay or switch, and the treatment chamber. Samples were
treated using 9 kV/cm, 720 Hz and different times; the response variable was the residual
vitamin concentration, in general all samples were analyzed by HPLC, additionally, samples
with E-carotene were also analyzed by UV/vis spectroscopy and samples with D-tocopherol
with IR spectroscopy [1-4].

RESULTS & DISCUSSION

It was found, that with 3, 5, 10 and 15 minutes of treatment there were 57%, 52%, 46% and
33% of residual E-carotene, respectively (p<0.05); however when E-carotene was in the
avocado paste there were no differences with the control, (p>0.05). With L-Ascorbic acid the
EF caused degradation of the vitamin at 0.188 min-1 and left 1.5% of residual vitamin after 20
minutes of treatment; while within avocado pulp no observed effect on the ascorbic acid.
Finally, the EF had no significant effect on standard of D-tocopherol alone and within avocado
paste (p > 0.05). These results are resumed in Table 1.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1619
 Table 1. Results of the EF treatment (9 kV/cm and 720 Hz) on three vitamins.

ȕ-carotene L-Ascorbic acid Į-tocopherol

ȕ-carotene Degradation/ min = L-Ascorbic acid residual
t(min) EF had no significant
residual (%) 0.188 /20 min = 1.5%
effect on standard of
3 57
D-tocopherol alone
5 52 while within avocado pulp no observed effect on
and within avocado
10 46 the ascorbic acid (p>0.05).
paste (p > 0.05).
15 33

It was also observed that E-carotene degraded itself even without EF treatment; samples left in
the equipment while it was set off for 5 and 15 min, had the vitamin degraded for ca 30 and
50% respectively, these observations confirm the labiality of this vitamin. Standard of ascorbic
acid is exponentially degraded (0.188 min-1) with time of EF treatment but in avocado paste
this vitamin remains after the EF treatment. The HPLC chromatograms of EF treated ascorbic
acid showed two more peaks indicating the degradation of vitamin C as a result of the EF
treatment. D-tocopherol was not affected by the electric field as standard or as added to the
avocado paste.

CONCLUSION
Electric field strongly degrades, as a function of treatment time, both E-carotene and ascorbic
acid when they are treated as pure compounds, the degradations follows well a kinetic
exponential decay, being faster the degradation of ascorbic acid. However when they are added
to the avocado paste both vitamins remains after treatment, indicating that all others
components of avocado paste apparently protect labile compounds, by absorbing the effect or
diluting it. D-tocopherol is not degraded as standard nor as added to the avocado paste,
indicating that this vitamin is stronger and better additive into foods as antioxidant.

REFERENCES
[1] Azevedo M.C., Rodriguez A.D., (2004). Con¿rmation of the identity of the carotenoids of tropical
fruits by HPLC-DAD and HPLC-MS. Journal of Food Composition and Analysis 17, 385–396.
[2] Che Man, Y.B., Ammawath, W., Mirghani, M.E.S. (2005). Determining D-tocopherol in re¿ned
bleached and deodorized palm olein by Fourier transform infrared spectroscopy. Food Chemistry 90:
323 –327.
[3] Elez-Martínez, P., Martín-Belloso, O. (2007). Effects of high intensity pulsed electric ¿eld processing
conditions on vitamin C and antioxidant capacity of orange juice and gazpacho, a cold vegetable
soup. Food Chemistry 102, 201–209.
[4] Rodriguez-Amaya, D.B., and Kimura, M. (2004). HarvestPlus Handbook for Carotenoid Analysis.
HarvestPlus Technical Monograph 2. United States of America.
[5] Torregrosa, F., Cortés, C., Esteve M., and Frígola, A. (2005). Effect of High-Intensity Pulsed Electric
Fields Processing and Conventional Heat Treatment on OrangeíCarrot Juice Carotenoids. J. Agric.
Food Chem. 53: 9519í9525

1620
Effect of Vacuum Impregnation Treatments to Improve Quality and Texture of Zucchini

(Cucurbita pepo, L.)

Elisabetta Occhino a, Isabel Hernando b, Paola Pittiaa,b

a
Department of Food Science, University of Teramo Mosciano S.Angelo (TE), Italy (ppittia@unite.it)
b
Departamento de Tecnología de Alimentos. Universitad Politecnica de Valencia Valenvia (Spain)

INTRODUCTION
Vacuum Impregnation (VI) is a non-destructive technology, used to introduce external liquids,
in the porous structures of food matrices favoured by the action of hydrodynamic mechanism
as promoted by pressure changes. The substitution of internal gases by a liquid phase of
adjustable composition allows formulation of a food by expeditious compositional
modifications of the solid matrix. Thus, while a change in the product composition occur, the
VI impregnated product could exert different quality (nutritional, sensory, textural, health)
properties depending on the functionality of the components of the VI solution. Widely
explored is the use of VI solutions made with sugars alone or in combinations with salt; in
recent times the addition of biologically active compounds is under study [1].
Zucchini (Cucurbita pepo, L.) is a wide known vegetable in the Mediterranean area at low
solute content, with a mild flavour. Its texture is typically soft but firm and changes
meaningfully during storage or when the vegetable undergoes to heating for dishes preparation.
Previous studies evidenced the potentiality to apply VI to zucchini [2] but no systematic
studies have been carried out yet. Aim of this study was to investigate the effect of VI by using
mixed solutions containing structuring ability components (maltodextrins, salt and CaCl2) with
on quality and microstructural properties of zucchini.

MATERIALS & METHODS

Slices of zucchini (0.5-cm thick) (Cucurbita pepo, L.) were subjected to VI treatments by using
a rotavapor equipment and the following constant process conditions: P: 50 mbar, vacuum time
(tv): 10 min; post-vacuum or relaxation time (tpv): 30 min; the ratio product:solution was equal
to 1:3.3. VI solutions with different solutes composition were investigated. In particular
maltodextrines (MD, DE 7.5-9, 10%), NaCl (0-5%) and CaCl2 (0-1000 mM) were used to
prepare simple (1 solute) or mixed (2 and 3 solutes) VI solutions After 2 min drainage,
samples were analysed for total solids, soluble solids, salt and [Ca] content, textural properties
(shear force, relaxation test), sensory and microstructure (Cryo-SEM). Mass balance (Solute
Gain/Loss, Water Gain/Loss) was also computed.

RESULTS & DISCUSSION

The composition of the VI solution affected mass transfer In particular the use of VI solutions
with NaCl (alone or in combination with MD and CaCl2) caused a significant water loss while
it did not occur when solutions prepared with MD and CaCl2, simple or mixed were used
(Table 1). A significant change in the textural properties (shear force and energy) was observed
in zucchini processed with VI solutions containing NaCl and MD; the presence of CaCl2 in the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1621
VI solution in combination with the other solutes, due to its structuring effect was able limit the
hardness loss and when alone even to determine an hardening effect (Figure 1).

Table 1. Water Gain/Loss (WG/L, g/g), Solid Gain/Loss ( SG/L, g/g), dry matter variation (% in respect
to the fresh no-VI product) of VI zucchini
Category WG/L (g/g) SG/L (g/g) Dry matter
(D%)
10% MD 0.144 0.018 10,86
2.5% NaCl -0.107 0.004 18,12
100 mM CaCl2 0.176 0.019 8,88
10% MD+2.5% NaCl -0.169 0.018 52,91
10% MD+2.5% NaCl -0.210 0.033 87,95
+ 100 mM CaCl2

Cryo-SEM analysis carried out on the zucchini processed with VI solutions with different
compositions was able to highlight a marked damage on the vegetable tissue due to a
dehydration effect when NaCl was used alone and on the contrary a structuring effect of the
CaCl2 with more thick cell walls and turgid cells. In the mixed solutions the presence of MD,
limited the damages induced by NaCl and CaCl2; intercellular spaces rich of solutes, turgid
cells and thick walls were in this case observed.

40
Shear Force
30
Shear Energy
20
10
0
' (%)

-10
-20
-30
-40
-50
10% MD 2.5% NaCl 100 mM CaCl2 10%MD+2.5% 10%MD+2.5%
NaCl NaCl+100 mM
CaCl2

Figure 1. Shear Force and Energy variation (%, in respect to the fresh no-VI product) of zucchini
processed by using VI solutions with different solutes composition

CONCLUSION
The use of VI mixed solutions made with MD, NaCl and CaCl2 to process zucchini allows to
favour solute and water gain while limiting textural and microstructural changes.

REFERENCES
[1] Zao Y., Xie J. 2004. Practical applications of vacuum impregnation in fruit and vegetable processing.
Trends in Food Science and Technology, 15, 434-451.
[2] Gras M., Vidal-Brotons D., Betoret N., Chiralt A., Fito P. 2002.The response of some vegetables to
vacuum impregnation Innovative Food Science & Emerging Technologies 3, 263_269.

1622
 Qualitative characteristics of sugar beet juices obtained in pilot extractor with pulsed

electric field (PEF) pre-treatment.

Kseniia Loginovaa,b, Eugène Vorobieva, Nikolai Lebovkab

a
Département de Génie Chimique, Université de Technologie de Compiègne, Centre de Recherche de
Royallieu, Compiègne, France, kseniia.loginova@utc.fr, eugene.vorobiev@utc.fr
b
Institute of Biocolloidal Chemistry named after F. D. Ovcharenko, NAS of Ukraine, Kyiv, Ukraine,
lebovka@gmail.com

INTRODUCTION
The conventional technology of sugar extraction from beetroot consists of a thermal
denaturation of sliced beetroot tissue followed by aqueous diffusion in hot water at 70–75 °C
[1]. However, thermally induced degradation of beet tissue, extraction of non-sucrose cell
components and formation of colorants decrease the juice purity and requires its further
purification. Recently, the alternative studies have shown the principal possibility of PEF-
assisted sugar extraction by cold or moderately heated water [2]; it was reported that the purest
juice was obtained after the cold diffusion. However, the confirmation of the encouraging
results obtained with PEF-treatment in batch extraction chambers is still needed to predict the
PEF efficiency on the industrial scale. This work is aimed at approach of the study of sugar
diffusion from sugar beets treated by PEF to the industrial conditions using special
countercurrent extractor. The parametric study is carried out for investigation of the effect of
electrical (intensity of PEF) and main extraction parameters (temperature, draft) on the sugar
diffusion kinetics, as well as on the juice and pulp characteristics.

MATERIALS & METHODS

The field-grown sugar beet roots (Beta vulgaris) were used throughout this study. Extraction
was carried out in a specially developed pilot countercurrent extractor. Construction and
detailed principle of this extractor were reported [3]. The total time of extraction was 70 min.
The temperature of diffusion varied in different experiments from 30 to 70 °C. In some
experiments, the draft value (diffusion juice to cossettes ratio) was varied between 120 and
90%, but typically it was fixed at 120%. Exhausted cossettes (pulp) after extraction were
pressed. The portions of extracted juice were regularly taken to measure the soluble solids
(°Brix), sucrose content, coloration, turbidity and colloids, as well as contents of proteins and
pectins. The portions of pulp were sampled for measuring the insoluble solids, °Brix and
sucrose content. Juice purity was calculated as: P = (Sucrose content/Soluble solids
content) ·100%. The intensity of electric field was typically fixed at E = 600 V/cm. One train
of pulses was used for PEF treatment. The train consisted of n pulses (n = 500) with pulse
duration ti = 100 ȝs and pulse repetition time ǻt = 5 ms. The total time of PEF treatment was
tPEF = 50 ms. In some experiments with higher diffusion temperature (60 °C), the value of E
varied between 600 and 100 V/cm.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1623
RESULTS & DISCUSSION
Fig. 1 presents the sucrose content and the purity of final juices obtained by diffusion of
untreated and PEF-treated cossettes. For the PEF-treated cossettes, the sucrose content in a
cold diffusion juice (30 °C) was slightly lower than in a hot diffusion juice (70 °C). However,
the purity of a cold diffusion juice obtained from the PEF-treated cossettes was not lower than
the purity of a hot juice (Fig. 1).
non-treated PEF treated
Sucrose content, °S

14 92
12 88

Purity, %
10 84
8
80
6
70 60 50 30 76 70 60 50 30
Temperature, °C Temperature, °C

Figure 1. Sucrose content and purity of juices extracted from non-treated and PEF treated cossettes
versus diffusion temperature.

The concentration of colloidal impurities was significantly higher in a “hot” juice than in a
“cold” juice. The measured concentrations of proteins in the extracted juices did not depend on
the temperature of extraction. From the other side, the increase of temperature above 50 °C
resulted in solubilization of pectins. Increase of extraction temperature from 30 to 70 °C
resulted in increase of turbidity by 10% (± 5%) and juice coloration by 27% (± 5%).
The pulp dryness was lower for cossettes exhausted at higher temperatures.
The sucrose content in diffusion juice was increasing with draft decrease for both studied
temperatures. However, the draft decrease resulted also in increase of sucrose losses in the
pulp. The optimal value of electric field intensity E was 257 V/cm for the diffusion at 60 °C.

CONCLUSION
The purity of diffusion juice obtained by cold extraction (at 30 °C) was not lower than the
purity of juice obtained by hot water diffusion at 70 °C; such juice had lower concentration of
colloidal impurities (especially, pectins) and lower coloration. Decreasing of draft permitted
better concentration of the extracted juice, but the cossettes were worse exhausted. Increase of
the temperature of the extracting liquid up to 50 or 60 °C permitted treatment of cossettes by
PEF with lower intensity. The pulp obtained by cold extraction of PEF treated cossettes had
noticeably higher dryness than the pulp obtained by conventional hot water extraction.

REFERENCES
[1] Poel, van der, P. W., Schiweck, H., Schwartz, T. 1998. Sugar Technology. Beet and Cane Sugar
Manufacture. Verlag Dr. Albert Bartens KG, Berlin.
[2] Jemai, A. B., Vorobiev, E. 2003. Enhanced Leaching from Sugar Beet Cossettes by Pulsed Electric
Field. Journal of Food Engineering, 59, 405–412.
[3] Loginova, K. V., Vorobiev, E., Bals, O., Lebovka, N. I. 2011. Pilot Study of Countercurrent Cold
and Mild Heat Extraction of Sugar from Sugar Beets, Assisted by Pulsed Electric Fields. Journal of
Food Engineering, 102, 340–347.

1624
 Modelling Microbial Load Reduction in Foods due to Ozone Impact

Elisabete M.C. Alexandre, Teresa R.S. Brandão, Cristina L.M. Silva

Centro de Biotecnologia e Química Fina - Escola Superior de Biotecnologia - Universidade Católica

Portuguesa, Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal (clsilva@esb.ucp.pt)

INTRODUCTION
Ozone, due to its powerful oxidizing effect, is one of the most potent disinfectant agents. In 1997 it
was recognised by U.S. Food and Drug Administration as a GRAS substance (i.e. Generally
Recognised as Safe) for use as a disinfectant or sanitizer in foods and food processing. The use of
ozone as a sanitizer is a challenging technology with potential application in the food industry.
When dissolved in water, ozone has been applied as a convenient washing treatment of fruits and
vegetables, promoting shelf life extension of food products [1]. Besides several studies assess the
ozone impact at microbial loads [2], scarce information is available on modelling the kinetic
behaviour of food contaminants. Such models may contribute to determine the extent to which the
process should be applied in order to improve safe standards and process design.
The main objective of this work was to study the impact of ozone in aqueous solution on the
following microbial loads: Listeria innocua in red bell peppers, total mesophiles in strawberries and
total coliforms in watercress. Modelling of microbial load reduction throughout treatment time and
due to ozone effect were also targets.

MATERIALS & METHODS

Strawberries (Fragaria ananassa D.), watercress (Naturtium officinale R.Br.) and red bell peppers
(Capsicum annuum L.) were purchased in a local market. Red bell peppers were pre-washed in
deionised water and cut in portions of 20 g. Each sample was artificially inoculated with 250 μL of
L. innocua with a contact time of 15 minutes.
Strawberries and watercress were not washed and did not suffer artificial contamination. Native
total mesophiles were evaluated in strawberry samples and total coliforms in watercress samples.
Ozone treatments were performed in a pilot plant. The gas was continuously incorporated by
bubbling in the water (at ~ 15 ºC) and the aqueous ozone concentration was 0.3 ppm. Samples were
immersed in ozonated water (80g / 30L) and were removed after different contact times. Simple
deionized water-washings (without ozone) were performed under identical conditions. L. innocua,
total mesophiles and coliforms enumerations were assessed using Palcam agar containing selective
supplement, Plate Count Agar and Violet Red Bile Agar, respectively.
A Weibull-based model was assumed for microbial load reduction throughout time [3]:
E
§ t·
§ N · ¨ ¸
log¨¨ ¸¸ 1  e© D ¹ (1)
© N0 ¹
where N is the microbial load (the index 0 indicates initial values) at time t; D is a scale parameter
(the reciprocal of the rate constant) and E is a shape index. The experimental data were fitted to the
previous equation, using SPSS 17.0 software.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1625
RESULTS & DISCUSSION
The impact of deionized-water and ozonated-water washings on L. innocua/red bell peppers
(N0=1.9x107 cfu/g), total mesophiles/strawberries (N0=5.6x107 cfu/g) and total coliforms/watercress
(N0=3.9 x108 cfu/g) can be observed in Figure 1. The effect of ozonated-water washings on
microbial loads reduction was higher than the one observed when a simple water-washing was
carried out, for all cases and treatment times considered. However, a substantial portion of the
microbial populations were reduced by water washing alone, and the presence of ozone generally
added an additional reduction of approximately 0.4 log-cycles for all times considered, and for the
combinations L. innocua/red bell peppers and total coliforms/watercress. The difference between
water and ozonated-water washings was particularly evident for total mesophiles/strawberries, and
for short contact times. As an example, for 1 minute of dipping, water-washings allowed 0.3 log-
cycles reduction and the presence of ozone added an additional reduction of 1.2 log-cycles.
The Weibull-based model was satisfactorily used in data fitting. Quality of regressions was assessed
by residual analysis (randomness and normality of residuals were verified) and on the coefficient of
determination, which varied from 0.71 to 0.96.
0.0 0.0 0.0
a) b) c)
-0.5 -0.5 -0.5

-1.0 -1.0 -1.0

Log(N/N0)
Log(N/N0)

Log(N/N0)
-1.5 -1.5 -1.5

-2.0 -2.0 -2.0

-2.5 -2.5 -2.5

a) b) c)
-3.0 -3.0 -3.0
0 1 2 3 4 0 1 2 3 4 0 1 2 3 4
Time (min) Time (min) Time (min)

Figure 1. Effect of water and ozonated-water washings on log-reductions of: (a) L. innocua/peppers, (b)
total coliforms/watercress and (c) total mesophiles/strawberries. Lines are model fits ( water; - - ozone).

CONCLUSION
Ozonated-water washings are more effective in reducing microbial loads of the fruits and vegetables
studied, when compared to simple water dipping. Total coliforms in watercress are less sensitive to
both deionized-water and ozonated-water washings.
A Weibull-based model was adequate in describing the reduction of microbial loads and may
contribute to design more effective sanitizing processes.

REFERENCES
[1] Ölmez H. & Akbas M.Y. 2009. Optimization of Ozone Treatment of Fresh-cut Green Leaf Lettuce.
Journal of Food Engineering, 90(4), 487-494.
[2] Akbas M.Y. & Olmez H. 2007. Effectiveness of Organic Acids, Ozonated Water and Chlorine
Dippings on Microbial Reduction and Storage Quality of Fresh-cut Iceberg Lettuce. Journal of the
Science of Food and Agriculture, 87(14), 2609-2616.
[3] Mafart P., Couvert O., Gaillard S. & Leguerinel I. 2002. On Calculating Sterility in Thermal
Preservation Methods: Application of the Weibull Frequency Distribution Model. International
Journal of Food Microbiology, 72, 107-112.

1626
 Use of organic acids on their own and in combination for decontamination of fresh
vegetables and herbs as an alternative to chlorine

Sami Buluta, Emel OgrasÕcÕb

a
Trakya University Food engineering department, Edirne, Turkey, samibulut@trakya.edu.tr
b
Trakya University Food engineering department, Edirne, Turkey, emelograsici@gmail.com

INTRODUCTION
Demand for fresh minimally processed fruits and vegetables increasing. However, fresh
produce poses a risk of food-borne outbreaks as pathogenic bacteria can contaminate raw
agricultural commodities through various pathways. Therefore, there is a need for effective
decontamination of fresh produce. Currently, chlorinated water is the most commonly used
wash agent to reduce the number of microorganisms on fresh produce. However, there has
been an increasing concern over the potential for formation of harmful by-products such as
organochlorine when fresh produce is washed in chlorine-based wash waters. In search of a
better and safer decontamination system, in this study, various organic acids were used on their
own and in combination to decontaminate some fresh vegetables and herbs. By studying a
wider range of organic acids, optimising the various parameters of washing or finding an
optimum combination for a mixture of organic acids could lead to a potential wash system that
is more efficient as an alternative to chlorine and could be used for organic produce
decontamination. Here, initial results for washing of parsley in spirit vinegar would be
presented.

MATERIALS & METHODS

Whole fresh parsley samples were purchased from local market and visibly soiled, spoiled or
yellow leaves were separated by hand (gloved). Spirit vinegar solutions were prepared by
adding calculated amount of spirit vinegar containing 20% acetic acid to sterile tap water. The
pH of the solution was monitored and recorded by a handheld pH meter. The temperature of
the solution was adjusted by using boiled water or submerging the solution bottles into an ice
water. Small plastic containers of 3 Litre capacity were used to soak the samples in solutions.
About 50 g of parsley samples were soaked into the solutions for set periods of time before
they were removed from the solution and placed into salad spinner to remove excess moisture.
The samples were then placed into stomacher bags and stored in the fridge (2-5ºC) until the
time of microbial analysis. Number of microorganisms on parsley before and after vinegar
wash was determined by total aerobic count (TAC) on Plate Count Agar (PCA) and results
expressed as logarithmic reduction. The effect of three main variables vinegar concentration
(2.0-7.0%), soaking time (0.5-7.5 min) and temperature (2.5-50°C) and their interactions were
studied by the surface response methodology employing Design-Expert V8 Software (Stat-
Ease, Inc. Minneapolis, USA) using central composite design.

RESULTS & DISCUSSION

Results showed that the solution temperature does not play significant role on reduction of
TVC. Temperatures that are optimum for growth of most microorganisms around (30ºC)

11th International Congress on Engineering and Food - Athens, Greece, 2011 1627
seemed to be less effective on TVC whereas lower (12ºC) or higher temperatures (40ºC) results
in slightly better results. As shown in Figure 1, increased concentration of vinegar resulted in
increased reduction in TVC. Increased contact time contributed to log reduction significantly at
higher vinegar concentrations whereas, at low concentrations increased contact time did not
contribute to log reduction in TVC. Results on effectiveness of vinegar (acetic acid) wash on
fresh produce and herbs are various [1-3]. These variations may be due to differences in the
types and quantities of test microorganisms, acid concentrations of treatment solutions and the
types of produce used.

1.4

1.2

1.0
Log Reduction

0.8

0.6

0.4

0.2

6.0
5.4
6.0
4.8
5.0
4.2
4.0
3.6
A: Concentration
3.0
B: Time 2.0 3.0

Figure 1. Experimental and simulated time temperature profile.

Response surface between acetic acid concentration, time of soaking and log reduction in parsley.

CONCLUSION
Results suggest that vinegar solutions at about 1% acetic acid concentration (5.0% vinegar)
results in similar reductions obtained by commercial chlorination wash (usually 50 ppm free
chlorine). More study is needed to understand the sanitising power of different organic acids on
their own and in various combinations.

REFERENCES
[1] Escudero, M. E., Velazquez, L., Di Genaro, M. S. & de Guzman, A. M. S. (1999). Effectiveness of
various disinfectants in the elimination of Yersinia enterocolitica on fresh lettuce. J Food Prot 62,
665-669.
[2] Sengun, I. Y. & Karapinar, M. (2005). Elimination of Yersinia enterocolitica on carrots (Daucus
carota L.) by using household sanitisers. Food Control 16, 845.
[3] Ruiz-Cruz, S. l., E. Acedo-Felix, M. D az-Cinco, M. A. Islas-Osuna, and G. A. Gonzalez-Aguilar.
2007. Efficacy of sanitizers in reducing Escherichia coli O157:H7, Salmonella spp. and Listeria
monocytogenes populations on fresh-cut carrots. Food Control 18:1383-1390..

1628
Use of a Weibullian model to characterize microbial inactivation in apple juice processed

with ultraviolet light

Mytilinaki Elisaveta, Guerrero Sandra a, b and Alzamora Stella. M a, b

a
Departamento de Industrias. Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires.
Ciudad Universitaria (1428) C.A.B.A., Argentina.e-mail: sguerrero@di.fcen.uba.ar
b
Member of Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.

INTRODUCTION
Thermal treatment constitutes the most extensively available method for the inactivation of
microorganisms in fruit juices to achieve the required 5-log reduction in number of the most
resistant pathogens but causes substantial changes in their flavor and nutritional quality. Minimal
processing of fruits may include many novel technologies aimed to minimize these changes and to
improve shelf life. The objective of this work was to evaluate and characterize the effect of UV-C
light on the survival of E. coli ATCC 35218 and a mixture of four E. coli strains (cocktail 1)
(ATCC 35218; 8738; 11229 and 25922); L.innocua ATCC 33090; S. cerevisiae KE 162 and a
mixture of five yeast strains (cocktail 2) (S. cerevisiae KE 162; C. parapsilosis ATCC 22019;
Z.bailii NRRL 7256; Z. rouxii ATCC 52519 and P.anomala NRRL 3668) in apple juice processed
in a continuous flow system.

MATERIALS & METHODS

Evaluated systems were filtered natural apple juice (FJ) (pH 3.5; 13.6°Brix) obtained in the
laboratory and a commercial type clarified apple juice (CJ) (pH 3.5; 13.4 ºBrix). Acidified peptone
water (PW) (pH 3.5) was used as control system. The UV-C device consisted of a 0.87 m-long glass
tube with a UV-C lamp (TUV-100W , 253.7 nm) leaving a annular flow space (equivalent
diameter, De= 0.01604m; transversal section for flow, S=0,00030m; volume=0.22 L). UV-C light
intensity was 14 kJ/m2 as determined by the iodure/iodate chemical actinometer [1]. Five hundred
(500) mL- inoculated systems were recirculated with a peristaltic pump (1 L/min; 28°C). Samples
were taken at preset time intervals during 15 minutes. Microbial populations were monitored by
plate count technique. Survival curves were generated from experimental data by plotting Log N/N0.
Weibullian type distribution of resistances model was fitted to experimental data [2].

RESULTS & DISCUSSION

S. cerevisiae KE 162; E.coli ATCC 35218 and L. innocua ATCC 33090 were successfully
inactivating using non thermal UV-C irradiation . The extent of microbial inactivation was
dependent on the microorganism and the treated media. E. coli was the most sensitive to the UV-C
irradiation treatment while S. cerevisiae, the least. A considerable level of inactivation of these
microorganisms (almost 5 log cycle reduction or greater) was obtained in commercial apple juice
(CJ) and in peptone water (PW) after 15 minutes of UV-C treatment. The profiles of inactivation
curves in PW were similar for all the studied microorganisms reaching between 4.3 (L.innocua) and
5.2 (E. coli) log reductions after 15 minute treatment. In CJ, the total inactivation ranged between
4.2 (S. cerevisiae) and 6.4 (E.coli) log reductions. However, there was a markedly lower efficiency
of UV-C disinfection (~ 3.1- 4.3 log red.) in filtered natural apple juice (FJ), probably due to the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1629
presence of colored compounds and pulp particles which caused poor UV-C light transmission. The
inactivation curves exhibited in all cases upward concavity, being almost biphasic. Experiments
using mixtures of strains were also carried out to analyze the change in the inactivation response due
to the presence of other strains.

Figure 1. a) Effect of UV-C irradiation on single and mixed strains in commercial apple juice. Experimental (points)
and fitted values derived from the Weibullian model (lines); standard deviation (I). (Š)E. coli ATCC 35218; (¡)
cocktail 1; (ƒ)S.cerevisiae KE 162; ( ) cocktail 2. Weibull frequency distributions of resistances corresponding to
survival curves. b) (ņ) E.coli ATCC 35218; (- -) cocktail 1. c) (ņ) S.cerevisiae KE 162; (- -) cocktail 2.

The cocktails of strains showed survival curves similar in shape (n < 1) to the individual strains. The
cocktail 1 was a little more sensible to the UV-C treatment than the single strain, E.coli ATCC
35218 but when the UV-C dose increased the inactivation of the cocktail decelerated remaining a
greater fraction of resistant population (Figure 1a). This change in the survival profile was
evidenced by its correspondent frequency distribution of resistances, which was more skewed to the
right, without mode and with higher spread of data (similar mean value and higher variance) (Figure
1b). S. cerevisiae and the cocktail 2 showed similar behaviour being more resistant than E.coli and
its cocktail. Their frequency distributions exhibited higher mean and variance values. Also, at doses
d 9.5 kJ/m2 (~10 min.), the cocktail 2 was more sensitive than the individual strain, S. cerevisiae KE
162 (Figure 1a,c).

CONCLUSIONS
This work contributed to address some limitations of novel technologies when applied to food
systems. UV-C radiation in a continuous flow arrangement was effective inactivating E. coli ATCC
35218; an E. coli cocktail; S. cerevisiae KE 162 and a yeast cocktail in peptone water and clarified
apple juice but had less inactivation effect in the filtrated natural apple juice. The Weibull type
distribution model evidenced differences between survival curves that did not surge by applying a
kinetic model. Further investigation is needed to elucidate the way UV-C treatments may be
performed to increase their efficiency and in order to evaluate the efficacy of the treatment in other
media or with other microorganisms of relevance.

REFERENCES
[1] Rhan R .1997. Potassium iodide as a chemical actinometer of 254 nm radiation: use of iodate as an
electron scavenger. Photochemistry and Photobiology, 66, 450- 455.
[2] Peleg M & Cole MB .1998. Reinterpretation of microbial survival curves. Critical Reviews in Food
Science, 38, 353-380.

1630
 Detection of pork freshness using NIR hyperspectral imaging

Douglas F. Barbina; Gamal ElMasrya; Da-Wen Suna; and Paul Allenb

Į
Food Refrigeration and Computerised Food Technology (FRCFT), School of Agriculture, Food Science
& Veterinary Medicine, University College Dublin, Dublin, Ireland (dawen-sun@ucd.ie).
b
Ashtown Food Research Centre, Teagasc, Dublin 15, Ireland.

INTRODUCTION
Freezing is recognized as a safe way to preserve meat for longer periods. However, meat shows
a gradual deterioration in quality attributes with frozen storage such as increased drip losses
and decreased protein extractability. When frozen pork is thawed, juices and micro nutrients of
the meat run out together with the ice water so the pork becomes dry and less tasty when
cooked. This drip loss also causes the loss of thiamine and folates amongst others, and
significantly downgrades appearance of meat. In order to prevent meat retailers from offering
thawed, imported frozen meat as fresh domestic ones, various methods have been proposed in
the past for identification of frozen and thawed meat [1]. The suitability of spectroscopic
methods for identification of frozen and thawed beef in the NIR and visible range has been
investigated [2-4]. It is advantageous to use near-infrared to provide more detailed spectral
information, since hyperspectral systems operating in the visible region (400-800 nm) have
considerably weaker band intensities when compared to the NIR range (900-1800 nm). In this
study, the overall objective was to investigate the ability of NIR hyperspectral imaging
technique for accurate and objective classification of pork samples according to its freshness.

MATERIALS & METHODS

A set of 15 pork chops (one inch in thickness) from the longissumus dorsi muscle (2 days post-
mortem) were imaged in the hyperspectral system. After imaging, the samples were vacuum-
packed and frozen in a commercial freezer. After 6 months, the samples were thawed for two
hours at room temperature and imaged again.
A pushbroom hyperspectral imaging system was used to take images from each sample before
freezing and after thawing. A region of interest (ROI) was selected comprising only the loin
eye area. The loin eye average spectral information was used for comparison between fresh and
frozen-thawed samples. For each image, a mean reflectance spectrum of the ROI was
calculated by averaging the spectral responses of all pixels in the ROI. In total, 30 mean
reflectance spectra were obtained, one for each sample fresh and after thawing.
Second derivative was performed for the extracted spectral data of the two conditions to
identify the main wavelengths that could explain the difference among samples. Principal
component analyses (PCA) were performed on the spectral data of pork samples and the
resulting loadings were then used to extract the useful information attributed to difference in
pork freshness.

RESULTS & DISCUSSION

The derivatives show the location of maximum spectral variance, where more important
features at 961, 1071, 1124 and 1147 were identified. These wavelengths were used to build a

11th International Congress on Engineering and Food - Athens, Greece, 2011 1631
PCA model to identify fresh and frozen-thawed pork meat with reduced data processing. The
first three principal components were responsible for 99.98% of variability of the data; the first,
second and third principal components variability were 99.22%, 0.61% and 0.15%,
respectively. The principal components method is able to differentiate pork samples based on
the reflectance values obtained from a reduced number of wavelengths.

Figure 1. (a) Second derivative spectra of fresh and frozen-thawed samples; (b) Score plot of first two
principal components for spectral data extracted from loin eye region of fresh and frozen-thawed samples.

The first three PCs obtained from the spectral data of pork samples were used for generation of
PC score images, as they accounted for more than 99.9% of the variance in the spectral data.
Visualization of pseudo-colour images allows for the further classification of samples based
either on their spectral information, as shown by the PC scores plot, or by the number of pixels
from the images. The model can be further enhanced by including a larger amount of samples.

CONCLUSION
The study demonstrated the potential of NIR hyperspectral imaging coupled with principal
components analysis for evaluating freshness of pork. The fresh and frozen-thawed pork
samples could be distinguished by the respective spectral attributes. Few selected wavelengths
could be potentially utilized by a multispectral imaging system for identifying fresh pork meat
in real time. Customers could benefit from a fast and non-invasive system to guarantee meat
freshness.

REFERENCES
[1] Ballin, N. Z.; Lametsch, R. 2008. Analytical methods for authentication of fresh vs. thawed meat - A
review. Meat Science, 80, (2), 151-158.
[2] Thyholt, K., Isaksson, T. 1997. Differentiation of frozen and unfrozen beef using near-infrared
spectroscopy. Journal of the Science of Food and Agriculture, 73 (4), 525-532.
[3] Downey, G.; Dominique Beauchêne, D. 1997a. Discrimination between fresh and frozen-then-thawed
beef m. longissimus dorsi by combined visible-near infrared reflectance spectroscopy: A feasibility
study. Meat Science, 45, (3), 353-363.
[4] Downey, G.; Beauchêne, D. 1997b. Authentication of Fresh vs. Frozen-then-thawed Beef by Near
Infrared Reflectance Spectroscopy of Dried Drip Juice. Lebensmittel-Wissenschaft und-Technologie,
30, (7), 721-726.

1632
Impact of non-thermal atmospheric pressure plasma on quality relevant food ingredients

Björn Surowskya, Falk Zülickea, Oliver Schlüterb, Dietrich Knorra

Björn Surowsky, Berlin University of Technology, Department of Food Biotechnology and Food Process
Engineering, Germany, bjoern.surowsky@tu-berlin.de
Falk Zülicke, Berlin University of Technology, Department of Food Biotechnology and Food Process
Engineering, Germany, falk_zuelicke@web.de
Oliver Schlüter, Leibniz-Institute for Agricultural Engineering Potsdam-Bornim, Germany,
oschlueter@atb-potsdam.de
Dietrich Knorr, Berlin University of Technology, Department of Food Biotechnology and Food Process
Engineering, Germany, dietrich.knorr@tu-berlin.de

INTRODUCTION
In the food industry, decontamination of fresh cut products is commonly achieved by the use of
chemicals such as hydrogen peroxide or hypochlorite. Chemical treatments are usually
attended by a loss of sensorial and nutritional quality and do still not achieve sufficient
microbial inactivation in some cases. New preservation methods are required to provide food
safety on the one hand and nutritional and sensorial quality on the other hand.

Non-thermal atmospheric pressure plasma processes might fulfil the demand for a gentle
treatment which provides food safety as well as nutritional and sensorial quality. Such plasmas
combine several antimicrobial effects which are mainly based on the generation of UV photons
and reactive species like radicals [1]. In this study, the impact of cold argon plasma on model
solutions and fresh non-pasteurized apple juice has been examined.

MATERIALS & METHODS

Model solutions (vitamin C, catechin, polyphenoloxidase (PPO)) as well as fresh apple juice
were treated by a non-thermal atmospheric pressure plasma jet.
Vitamin C was dissolved in meta-phosphoric acid, catechin in a methanold/H2O mixture and
PPO in a pH 6.5 buffer solution. The amounts were based on their occurrence in apple juice.
Royal Gala apples were obtained from a local supermarket (Kaisers-Tengelmann, Berlin,
Germany). After the extraction (Gastroback Design Juicer Advanced 800), the juice was
centrifuged, filtered, filled into PE bottles and put into a freezer (-18 °C).

After thawing the samples, 2 ml of each were filled into small glass bottles and treated with the
plasma jet. The plasma was generated by exposing the feeding gas (argon 5.0) to a 1.1 MHz
electric field at the inner needle electrode of the jet. The gas composition (pure argon, argon +
0.1 % O2, argon + 0.01 % O2) and the treatment time (0 to 120 s) were varied. Immediately
after each treatment, the samples were filled into ice-cooled micro-centrifuge tubes.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1633
The concentration of vitamin C was observed by HPLC, equipped with a UV detector (Knauer,
Germany) and a 250 x 4 mm Hypersil ODS column (Knauer, Germany), using a
tetrabutylammonium hydrogen sulfate/dH2O/methanol mixture as eluent [2].
The total amount of phenolics was determined spectrometrically according to the method of
Folin-Ciocalteu [3].
By adding catechol to PPO containing samples, an oxidation to catechin takes place. The
resulting change in absorption was spectrometrically observed at a wavelength of 420 nm for
60 s [4].

RESULTS & DISCUSSION

The results of the treatment of fructose free model solutions showed a relatively uniform
image. Pure argon plasma showed the strongest effects: PPO as well as vitamin C and phenolic
compounds were harmed. After a treatment time of 120 s, the contents of vitamin C and
phenolic compounds decreased up to 45 % and 10 %, respectively. With a reduction up to 95
%, the PPO activity was the most affected property.
In general, the deterioration of these compounds was reduced by addition of fructose prior to
the plasma treatment. Neither the contents of vitamin C and phenolics nor the fructose itself
were affected, whereas the reduction of PPO activity dropped from 95 to 50 %. In the case of
PPO, it was possible to increase the inactivation from 50 to 70 % by adding 0.01 % oxygen to
the feeding gas.
After treating the complex apple juice matrix, the contents of vitamin C as well as the phenolic
compounds did not show significant differences compared to the control samples. The PPO
activity was reduced up to 20 % after treating the samples for 120 s regardless of the used gas
composition.

CONCLUSION
It can be concluded, that the complex food matrix has a protective effect on ingredients like
Vitamin C, phenolics and PPO. By means of the model solutions, it has been revealed that
carbohydrates like fructose are jointly responsible for this. Based on successful experiments
concerning the bactericidal effects of non-thermal atmospheric plasmas, these first promising
results demonstrate that cold plasmas could be suitable for enhancing the safety of perishable
foodstuff without affecting their nutritional quality too much.

REFERENCES
[1] Moisan, M.; Barbeau, J.; Moreau, S.; Pelletier, J.; Tabrizian, M.; Yahia, L’H. (2001): Low-
temperature sterilization using gas plasmas: a review of the experiments and an analysis of the
inactivation mechanisms. International Journal of Pharmaceutics 226, 1–21.
[2] Rückemann, H. (1980): "Methoden zur Bestimmung von L-Ascorbinsäure mittels Hochleistungs-
Flüssigkeitschromatographie (HPLC)." Zeitschrift f. Lebensmitteluntersuchung u. Forschung A,
171, 357-359.
[3] Singleton, V. L.; Rossi, J. A. (1965): Colorimetry of Total Phenolics with Phosphomolybdic-
Phosphotungstic Acid Reagents. In: Am. J. Enol. Vitic. 16, Nr. 3, S. 144-158.
[4] Siriphanich, J; Kader, A. A. (1985): "Effects of CO2 on total phenolics, phenylalanine ammonia
lyase and polyphenoloxidase in lettuce tissue." Journal of the American Society of Horticultural
Science, 110(2), 249-253.

1634
 Effect of pulsed light and ascorbic acid/CaCl2 dipping on rheological properties of fresh-

cut apples

Paula L. Gómez1,3, Daniela M. Salvatori2,3 and Stella M. Alzamora1,3

1
Departamento de Industrias. Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires,
Ciudad Universitaria, 1428 Buenos Aires, Argentina. (e-mail: alzamora@di.fcen.uba.ar )
2
Facultad de Ingeniería, Universidad Nacional del Comahue, Neuquén, Argentina.
3
CONICET, Buenos Aires, Argentina.

INTRODUCTION
Pulsed light (PL) treatment has emerged in recent years as an alternative to thermal treatment
for inactivation of pathogenic and spoilage microorganisms. It involves the use of intense and
short-duration (1 μs to 0.1 s) pulses of broad spectrum light of wavelength ranging from UV to
near-infrared (200 to 1,100 nm) [1, 2]. Its use has been approved by the FDA for the
decontamination of food and food surfaces [3]. The significant microbial reduction in very
short treatment times, the limited energy cost, the lack of residual compounds, and its great
flexibility are some of the major benefits claimed for this technique [4].
The literature on the application of pulsed light in foods is still scarce, especially regarding
minimally processed fruits and vegetables. The potential application of this technology in
minimal processing of fruits should not be only evaluated on its ability to contribute to food
safety but also on its influence on produce quality (color, texture, taste and aroma). In previous
works it has been reported that irradiation at higher doses produce an increase in the surface
browning, which can be prevented by application of a suitable antibrowning agent prior to
irradiation. The aim of this work was to evaluate the changes on linear viscoelastic properties
of fresh-cut apples irradiated with pulsed light (PL) subjected or not to an antibrowning
pretreatment.

MATERIALS & METHODS

Apple slices (3 cm diameter and 0.6 cm thickness) were dipped into a 1% (w/v) ascorbic acid +
0.1 % (w/v) calcium chloride solution for 5 min at 4ºC. The PL treatments were performed
with a RS-3000B Steripulse-XL system, which produced polychromatic radiation (200 to 1100
nm) and generated high intensity pulsed light. Samples were exposed to irradiation for 60 s at
10 cm from the lamp (fluence: 72 J/cm2). Slices irradiated with and without the antibrowning
pretreatment were compared to a control throughout a week of storage at 4-5ºC. Dynamic
oscillatory shear and creep/recovery tests were performed at 0 and 7 days. Creep behavior was
fitted with a mechanical model (a spring, two Kelvin-Voigt elements and a dashpot). Light
microscopy observations were made after processing and storage.

RESULTS & DISCUSSION

Storage moduli (G’) of apples exposed to PL were lower than those of the fresh fruit, both at 0
and 7 day. The decrease was more pronounced when apples were pretreated with the
antibrowning solution (data not shown). The instantaneous compliance (J0) and the retarded
compliance (J2) increased after 7 day storage in samples irradiated (with and without AD),

11th International Congress on Engineering and Food - Athens, Greece, 2011 1635
denoting that these tissues became more deformed. Rheological response of apple tissue was
correlated with microstructure and ultrastructure features.

Table 1. Viscoelastic parameters* for fresh and treated apple tissues stored at 5ºC. PL: pulsed light; AD:
antibrowning dipping.
J0 J1 J2 KN
Time
Treatment (1/Pa) (1/Pa) (1/Pa) O1 (s) O2 (s) (Pa.s)
(day)
(x 106) (x 106) (x 106) (x 10-8)
Control 3.1(0.9) 1.4(0.7) 0.7(0.3) 26.4(3.9) 2.6(0.5) 0.7(0.5) A

AD 3.1(0.8) 4.3(3.2) 0.5(0.2) 32.9(12.2) 3.1(1.1) 1.9(1.2) B

0
PL 4.9(1.1) 1.4(0.3) 0.9(0.2) 26.2(8.1) 2.4(0.6) 0.8(0.3) CDA
1.2
AD + PL 3.5(0.5) 0.6(0.1) 35.0(19.1) 2.1(0.6) 1.5(0.8) DA
(0.5)
1.1
Control 4.1(0.9) 1.9(1.8) 30.3(13.4) 2.6(0.6) 1.7(0.8) D
(0.4)

1.4
AD 5.2(1.4) 0.9(0.3) 19.1(5.7) 1.9(0.7) 0.8(0.6) CDA
(0.7)
7
PL 8.2(4.4) 2 (1) 2(1) 22.8(3.8) 2.1(0.6) 0.6(0.2) E

3.1 2.2
AD + PL 13(5.2) 21.3(3.9) 2.6(0.9) 0.4(0.2) E
(1.8) (0.9)
Results were expressed as mean followed by the standard deviation into brackets.*Parameters derived by fitting the
average compliance data from the creep phase. Different uppercase letters indicates significant differences between
treatments (p<0.05).

CONCLUSIONS
Pulsed light at the dose tested induced changes in viscoelastic properties of apple tissue that led
to both a loss of stiffness and higher capacitance values. These changes were increased by
ascorbic acid/CaCl2 treatment and during storage. Due to the low penetration of PL irradiation,
changes in viscoelasticity would be given mainly on surface region of irradiated samples.

REFERENCES
[1] Dunn J, Ott T. & Clark W (1995) Pulsed-light treatment of food and packing. Food Technology, 49,
95-98.
[2] Gómez-López VM, Ragaert P., Debevere J & Devlieghere F. (2007) Pulsed light for food
decontamination: a review. Trends in Food Science & Technology, 18, 464-473.
[3] FDA (1996) Code of Federal Regulations. 21CFR179.41.
[4] Oms-Oliu G, Aguiló-Aguayo I, Martín-Belloso O & Soliva-Fortuny R (2010) Effects of pulsed light
treatments on quality and antioxidant properties of fresh-cut mushrooms (Agaricus bisporus). Postharvest
Biology and Technology, 56, 216-222.

1636
 Modeling a pasteurization process of clarified apple juice based on pulsed ultraviolet

light

I. Kasaharaa, P. Grogga, L. Aguilarb

a
Escuela de Alimentos, Universidad Católica de Valparaíso, Valparaíso, Chile
(ikasahar@ucv.cl)
b
Laboratorio de Fotofísica y Espectroscopía Molecular, Universidad Católica de Valparaíso,
Chile (luis.aguilar@ucv.cl)

INTRODUCTION
Nowaday there is an increasing demand on more natural and minimally processed food.
Nevertheless this may result in an increase of consumer health risks due to a raw condition of
such foods and this has called attention from health authorities CFSAN-FDA [3]. Pulsed
ultraviolet (PUV) light seems to be a promising alternative to process food since it represents a
non thermal technology and does not leave chemical residues in the product Barbosa-Canovas
[1], Bialka [2], Demirci [4]. Extensive research have been carried out to determine relationship
existing between process parameters and the effect on microorganisms lethality and kinetics on
traditional thermal processes, such as sterilization or pasteurization, whereas only few works
have been published related to food processing based on PUV.
The aim of this work was to determine the lethality of microorganisms in clarified apple juice
when subjected to a pasteurization process based on a pulsed ultraviolet light treatment at
different conditions and establish relationship between inactivation and process parameters.

MATERIALS AND METHODS

The elimination of E. coli and A. acidoterrestris microorganisms on clarified apple juice by
mean a pasteurization process based on a pulsed ultraviolet (PUV) light treatment was studied.
PUV light was generated in an excimer laser source (Lambda Physik Compex 110). Survival
microorganisms before and after exposing juice samples to PUV light at different doses were
counted as colonies forming units. In order to establish the effectiveness of different levels of
energy, experimental data were adjusted to a Weibull model, according to equation proposed
by van Boekel (2002).
Samples of apple juice were inoculated separately with both microorganisms, and then exposed
to PUV light into quartz cells under static regimen and into a flow cell for a dynamic regimen.

RESULTS & DISCUSSION

Figures 1 present results related to the elimination of E. coli after PUV light pasteurization
process under a static and a dynamic regimen, respectively. Figure 1 shows that Weibull model
fit adequately experimental data. Besides, Figure 1(a) that represent a static condition, shows a
concave curve shape which in turn relates to Weibull model factor ß < 1, which indicates that
under a static regimen surviving cells have less probability to be inactivated while PUV light
dose is increased. On the contrary, Figure 2(b) which represents results obtained on a dynamic

11th International Congress on Engineering and Food - Athens, Greece, 2011 1637
regime, presents a concave curve shape and so factor ß > 1, indicating that microbial cells seem
to be more susceptible to be destroyed as PUV light dose is increased.
On the other hand, results obtained on elimination of vegetative cells and spores of A.
acidoterrestris, indicate that both forms show curves with ß < 1, therefore it could be assumed
that vegetative and latent forms have less probability to be inactivated while PUV light dose is
increased.

(a) (b)

Figure 1. Inactivation of E. coli in clarified apple juice after pulsed ultraviolet light pasteurization
process at different energy doses. Experimental data adjusted according Weibull model. (a) Static
regime, (b) flow regime

CONCLUSION
It is concluded that microorganisms’ destruction in clarified apple juice by mean a pulsed
ultraviolet pasteurization process essentially depends on energy dose. Besides, microbial
inactivation seems to depend also on apple juice clearness and soluble content. Experimental
data fit satisfactory well curves produced by Weibull model, either for inactivation of
vegetative cells of A. acidoterrestris and E. coli or spores form in the case of A. acidoterrestris.

REFERENCES
[1] Barbosa-Canovas G., Palou E., Pothakamury U. and Swanson B. 1998. Nonthermal Preservation of
Foods. 3rd Edition. Marcel Dekker Inc., New York, USA.
[2] Bialka K. & Demirci, A. 2008. Efficacy of pulsed UV-Light for decontamination of Escherichia coli
O157:H7 and Salmonella spp on raspberries and strawberries. Journal of Food Science 73(5), 201-207.
[3] CFSSAN-FDA 2000 Kinetics of microbial inactivation for alternative food processing technologies:
pulsed light technology. Center for Food Safety and Applied Nutrition, Food and Drug Administration.
http://vm.cfsan.fda.gov/comm/ift-puls.html
[4] Demirci A. 2002 Novel processing technologies for food safety. J. Assoc. Food Drug Officials 66(4),
1-8.

1638
 Encapsulation of Lactobacillus paracasei using Spray Gun technology

Maribel Jiménez a, Esmeralda Jiménez a, Ebner Azuara a, Guadalupe Luna b, Cesar I. Beristain a

a
Instituto de Ciencias Básicas, Universidad Veracruzana Xalapa ,Veracruz, México
(maribjimenez@uv.mx).
b
DEPI, Instituto Tecnológico de Orizaba, Orizaba, Veracruz, México

INTRODUCTION
Probiotic bacteria are defines as “live microorganisms which, when administered in adequate
amounts, confer a health on the host”. The ability of microorganisms to survive and multiply in
the host strongly influences their probiotic benefits. However the growth activity of probiotic is
affected by conditions such as pH, temperature, oxygen and the other factors [1]. A promising
solution to this problem is the encapsulation; the encapsulation is a process in which the cells
are retained within an encapsulating matrix [2]. The spay gun is a technique novel in the
pharmaceutical industry, this technique is cheap, uses room temperature is beneficial to
maintain the viability. On the other hand too keep the viability and extend their storage shelf
life is convenient to convert the capsules into drying employing techniques such as fluidized
bed drying [3]. The main objective if this study is to evaluate the application a novel technique
spray gun to encapsulating as well as the application of fluidized bed drying to produce dry
capsules of long shelf life, highly viable probiotic from Lactobacillus paracasei.

MATERIALS & METHODS

A lyophilized culture of Lactobacillus paracasei was cultured for 12 h at 37º C.
encapsulation procedure. Beads were a produced using a modified encapsulation method
originally by Sheu and Marshall [4] and Sultana et al. [5]. A 2 % alginate mixture was
prepared containing 2 % maize starch and 0.1 % culture. The mixture was added into 10 %
canola oil. The operating conditions of spray gun were as follows; spray pressure, 30 PSI,
nozzle diameter, 1.7 mm. Then the solution was sprayed on the 3% CaCl2 solution. For
obtained dry capsules used fluidized bed drying were as follows; fluidization velocity, 99 m3/s,
inlet temperature 30 ºC , the viable cells were counted according to the standard plate method.

RESULTS & DISCUSSION

The initial physical properties of capsules prepared and obtained by spray gun had a initial bulk
density was 0.458 g/cm3, compact density was 0.67 g/cm3, and particle density was 0.76
g/cm3. This important because, physical properties are directly related to the quality and
utilization of food products, causing changes in their structure and porosity. The physical
characteristics of the individual particles are mainly determined by the encapsulating material
and physical properties and indirectly, these properties can provide predictions about storage
stability. The initial moisture was 86 % and after 30 minutes of drying the moisture was 2.3 %.
The physical characteristics on the individual particles are mainly determined by the material
from which are encapsulated.
Powder property measurement is important because these properties intrinsically affect powder
behavior during storage, handling and processing such as transportation, mixing and
packaging. The viability of the solution before encapsulation was 7.10 X 109cfu, and after

11th International Congress on Engineering and Food - Athens, Greece, 2011 1639
encapsulation by Spray Gun was 2.40 X 108 CFU, which represent 3.38 % of the viability with
respect to the initial solution.
These results were similar to report by Semyonov et al. [6], when they used coacervation. The
vialibility of microorganism decreases with increased drying time, reaching values of 53 x105
corresponding to 2.2 % from initial viability (Figure 1).
1 0 0

8 0

6 0
% Viability

4 0

2 0

0
0 5 1 0 1 5 2 0 2 5 3 0 3 5

T im e (m in )
Figure 1.Viability of Lactobacillus paracasei encapsulating by spray gun and by fluidized bed drying.

One principal criterion for the stability of capsules is the moisture content. The higher viability
was when the capsules had moisture 36 %. When the capsules was dry the viability decreases
obtaining a major viability when the moisture content of capsules was high, the viability after
30 minutes of drying was 5.40x105, this viability was similar to reported by Semyonov et al [6]
when they used spray freeze to drying method and coacervation method of encapsulation

CONCLUSION
The present study demonstrate that fluidized bed drying is an appropriate process to generate
dried capsules of defined dimensions containing probiotic bacteria L. Paracasei, that retain
viability during the encapsulation by spray gun. We conclude that encapsulating by spray gun
and drying by fluidized bed drying are a good option to maintain viable L. paracasei to
incorporate new food systems.

REFERENCES
[1] Annan N.T., Borza N.T. & Trueistrup Hansen L. 2007. Encapsulation in alginate-coated gelatine microspheres
improves survival of the probiotic Bifidobacterium adolescentis 15703T during exposure to simulated
gastrointestinal conditions. Food Research International, 41, 184-193
[2] Homayouni A., Azizi A., Ehsani M.R., Yarmand M.S. & Razavi S.H. 2008. Effect of microencapsulation and
resistant starch on the probiotic survival and sensory properties of symbiotic ice cream. Food Chemistry, 111, 50-
55
[3] Anal A. & Singh KY. 2007. Recent advances in microencapsulation of probiotics for industrial applications and
targeted delivery. Trends in Food Science and Technology, 5, 240-251.
[4] Sheu T. Y. & Marshall, R.T., 1993. Improving survival of culture bacteria in frozen desserts by microentrapment.
Journal of Dairy Science, 76(7), 1902-1907
[5] Sultana K., Goodward, G., Reynolds N., Arumugaswamy R., Peiris P. & Kailasapathy K, (2000). Encapsulation
of probiotic bacteria with alginate-starch and evaluation of survival in simulated gastrointestinal conditions and in
yoghurt. International Journal of Food Microbiology, 62, 47-55.
[6] Semyonov D., Ramon O., Kaplun Z., Levin-Brener L., Gurevich N. & Shimoni E. 2010. Microencapsulation of
Lactobacillus paracasei by spray freeze drying. Food Research International, 43, 193-202.

1640
 Concentration of a vegetal enzymatic extract by microfiltration

Teles, A.S.C.a, Terzi, S.C.b, Silva, L. F. M.b, Gomes, F.S.b; Moraes, I.V.M.c; Egito, A.S.d; Cabral,
L.M.C.b; Matta, V.M.b

a
Central State University of West Zone, Rio de Janeiro, Brazil (aline_cascaes@yahoo.com.br)
b
Embrapa Food Technology, Rio de Janeiro, Brazil (vmatta@ctaa.embrapa.br)
c
Embrapa Tropical Agroindustry, Fortaleza, Brazil (ingrid@cnpat.embrapa.br)
d
Embrapa Goats and Sheep, Sobral, Brazil (egito@cnpc.embrapa.br)

INTRODUCTION
Milk clotting for cheese producing is usually performed using enzymes from animal sources,
and the most common is chymosin. However, its low availability and the increase in cheese
production have contributed to the lack of chymosin in world market, resulting in the search for
new sources, like the vegetal ones [1, 2]. An example of such application is the use of proteases
from vegetal that will also attend people with some religious or philosophical demands. Egito
et al. [2] have shown that sunflower extracts can be used for cheese production without
unintended flavor and that they have the same site of hydrolysis of chymosin when in presence
of casein. The objective of this study was to evaluate the concentration by membrane processes
of a vegetal proteolytic enzymatic extract obtained from sunflower seed.

MATERIALS & METHODS

The enzymatic raw extract was obtained according to the following steps: disintegration of
sunflower seed in a pilot plant mill, homogenization in an industrial blender, aqueous
extraction in brine solution containing 1% (w/v) NaCl during 18 hours under refrigeration and
centrifugation in a basket centrifuge (Bellinox, Brazil) at 35g, using a 150μm nylon screen
filter medium. The raw extract was microfiltered using ceramic membranes with 1.0 μm pore
size at 25oC and 2 bar of transmembrane pressure. Permeate flux was determined along the
process and samples were collected from feed (raw extract), permeate and retentate
(concentrated extract), for determination of total proteins, proteolytic activity and coagulation
unit.

RESULTS & DISCUSSION

The permeate flux behavior of microfiltration process is showed in Figure 1 where it is
possible to observe its stability along more than 200 minutes. The average flux value was 33
L/hm2 and a volumetric concentration factor equal to 5.0 was obtained in the process.
The results of total protein, proteolytic activity and coagulation unit are presented at Table 1.
As expected the coagulation unit was lower in the retentate fraction (4.72 CU/mL) than in the
feed (10.93 CU/mL) and no milk coagulation was verified with the permeate sample. No
proteolytic activity was detected in the permeate fraction and the total protein content was 0.29
mg/mL. Proteolytic activity has increased 1.5 times while total protein content has increased
about four fold. Both total protein (3.7 times) and enzymatic activity (1.5 times) were
concentrated in retentate fraction although they were not in the same range of the concentration
factor, probably due to the protein denaturing and enzyme inactivation. Neves et al. [3], when

11th International Congress on Engineering and Food - Athens, Greece, 2011 1641
studying the squid protein concentration by ultrafiltration, has attained a lower concentration
factor (1.8) although they have gotten to concentrate total proteins at the same range.

Figure 1. Permeate flux behaviour of sunflower extract microfiltration.

Table 1. Total protein, proteolytic activity and coagulation unit of the three fractions during
microfiltration of sunflower enzymatic extract
Parameter Total protein Proteolytic activity Coagulation unit
(mg/mL) (U/mL) (CU/mL)
Feed (raw extract) 1.69 ± 0.17 1.43 ± 0.07 10.93 ± 0.09
Permeate 0.29 ± 0.02 0 nc
Retentate 6.21 ± 0.24 2.18 ± 0.20 4.72 ± 0.01
nc – no coagulation

CONCLUSION
The obtained results suggest a potentiality for the concentration of the sunflower seed protease
by microfiltration, although this process needs further studies to reduce enzymatic activity
losses.

REFERENCES
[1] Cavalcanti M.T.H., Teixeira, M.F.S., Lima Filho, J.L. & Porto, A.L.F. 2004. Partial purification of
new milk-clotting enzyme produced by Nocardiopsis sp. Bioresource Technology, 93, 29–35.
[2] Egito A.S., Girardet J.M., Laguna L.E., Poirson C., Mollé D., Miclo L., Humbert G. & Gallard J.L.
2007. Milk-Clotting Activity of Enzyme Extracts from Sunflower and Albizia Seeds and Specific
Hydrolysis of Bovine k-casein. International Dairy Journal, 17, 816-825.
[3] Neves L.C, Cabral L.M.C, Stephan M.P., Leite S.G.F. & Matta V.M. 2006. Recovery of proteins from
residual brine of squid processing. Alimentaria, 119-123.
Acknowledgments: Embrapa and FUNCAP, for project funding, and Huberto Paschoalick for the seeds
supplying.

1642
 Fresh produce decontamination by an atmospheric pressure plasma-jet

a
Baier, M.; a Görgen, M.; a Fröhling, A.; a Geyer, M.; a Herppich, W.B.; b Ehlbeck, J.; c Knorr, D.;
a
Schlüter, O.

a
Leibniz-Institute for Agricultural Engineering, Potsdam, Germany (mbaier@atb-potsdam.de)
b
Leibniz-Institute for Plasma Science and Technology, Greifswald, Germany (ehlbeck@inp-
greifswald.de)
c
Technische Universität Berlin, Berlin, Germany (dietrich.knorr@tu-berlin.de)

INTRODUCTION
Developments of new atmospheric pressure plasma sources which allow generation of non-
thermal plasma at less and less temperatures approaching room temperature appear suitable to
treat heat-sensitive food surfaces. Over the last decade first attempts were conducted. The
antibacterial efficiency of a needle-to-plate-system was tested on apple juice and resulted in a
7 log units reduction of Escherichia coli O157:H7 [1]. Corresponding to a 2 log units
inactivation a helium-oxygen plasma could be applied on bell pepper without significant
effects of discoloration [2]. Using one atmosphere uniform glow discharge plasma samples
were treated in a remote exposure chamber. Strains of E. coli O157:H7, Salmonella and
Listeria monocytogenes on apples, cantaloupe, and lettuce, respectively, were reduced by at
least 2 log units [3]. A gliding arc plasma inactivated E. coli O157:H7 and Salmonella Stanley
on the surface of apples by 3 log units [4]. The efficacy of a plasma-jet on a pathogenic strain
and different spoilage microorganisms was examined on the pericarps of mango and melon [5].
The aim of this study was an alignment of antibacterial efficiency and quality assurance.
Different process parameters were varied to gain enhanced inactivation. Successful parameter
combinations were applied on Lambs lettuce and their suitability was monitored using
Chlorophyllfluorescence-Image-Analysis.

MATERIALS & METHODS

The used plasma device was an atmospheric pressure plasma-jet with argon as process gas and
a gas flow of 3 – 7 slm (standard liters per minute). The distance between sample and tip of the
plasma-jet was set at 18 mm. At 230 V and 50 Hz the jets power intake was at 8 W.
Escherichia coli was prepared using a two steps cultivation (pre-culture 24 h, 37 °C, main
culture 18 h, 37 °C, shaking at 125 rpm). 5*106 E. coli cells were placed on polysaccharide gel
discs (A=1 cm2), dried, and plasma-treated with varied parameters. Bacteria were resuspended
in 1 ml phosphate buffered saline, serially diluted and plated on ST-I agar plates.
Leaves of Lambs Lettuce were harvested and placed into sample holders, stored in a dark place
for 5 min to stop its photosynthetical activity. The chlorophyllfluorescence of the leaves was
measured before and immediately after treatment to assess the plasma´s impact on the tissue.

RESULTS & DISCUSSION

The inactivation kinetics show the impact of varied process parameters on the antibacterial
efficiency. The highest voltage applied resulted in highest inactivation of 3 log cycles.
However, within the first minute, treatment at 45 V remained about 1 log cycle less effective
than 35 V. Consequently, no direct relation of applied power and efficiency could be derived.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1643
Since treatment at maximum voltage of 65 V resulted in highest inactivation after 90 s and
120 s, this setting was chosen for further experiments. Altering the flow rate, the lowest value
of 3 slm showed 0.5 log cycles less inactivation after 2 min. The high gas flow of 7 slm
showed less effectiveness within the first 90 s and evened up with the medium gas flow within
the last 30 s. Achieving 3 log cycles inactivation the medium gas flow rate of 5 slm turned out
to be the most efficient setting. A considerable improvement was achieved when modifying the
gas composition. Small additions of 0.01 % and 0.05 % O2 to argon did not increase or even
decrease the efficacy. The addition of 0.1 % O2 resulted in inactivation of 3 log cycles after 1
min and 5 log cycles after 2 min treatment time.
As the average temperature (Tavg) did not exceed 15 °C, thermal effects in the inactivation
process can be excluded.
Following the microbiological tests, the most efficient parameter combination of 65.0 V,
5.0 slm flow rate, and a 0.1 % O2-addition was chosen to treat the lamb´s lettuce. The
measured variable Fv/Fm shows the maximum photosynthetic activity (exciton transfer
efficiency) which corresponds to the integrity of the photosynthesis apparatus. At a distance
between sample surface and tip of the plasma-device of 11 mm, all treatments from 20 s to 60 s
resulted in severe loss of up to 50 % Fv/Fm. Selecting a distance of 13 mm the initial decrease
has been reduced by almost 50 % and the impact of treatment duration appears more clearly. A
further increase to a distance of 18 mm between lettuce leaves and the tip of the plasma-device
did not lead to any effects after 20 s, 30 s, and 40 s. After 60 s treatment time Fv/Fm did not fall
below the characteristic level of 0.65 where alterations can be regarded as reversible. Until the
end of 72 h of observation the samples entirely recovered to the initial level before plasma
treatment.

CONCLUSION
The investigated plasma treatment resulted in a 5 log cycle inactivation of E. coli 147 after 2
min treatment time. The lamb´s lettuce remained intact after 60 s treatment time applying
optimised process parameters, which corresponds to a 3.0 log cycle inactivation. Further
experiments will be conducted to allow extended treatment times in plasma application for
fresh produce.

REFERENCES
[1] Montenegro RR (2002) Inactivation of E. coli O157:H7 using a pulsed nonthermal plasma system.
Journal of Food Science, 67(2), 646-648.
[2] Vleugels M., Shama G., Deng X.T., Greenacre E., Brocklehurst T., Kong M.G. (2005). Atmospheric
plasma
inactivation of biofilm-forming bacteria for food safety control. IEEE Transactions on Plasma
Science 33, 824-
828.
[3] Critzer, F.J., Kelly-Winterberg, K., South, S.L., Golden, D.A., 2007. Atmospheric plasma inactivation
of foodborne pathogens on fresh produce surfaces. Journal of Food Protection 70 (10), 2290–2296.
[4] Niemira B.A. and Sites J., 2008. Cold plasma inactivates Salmonella Stanley and Escherichia coli
O157:H7 inoculated on golden delicious apples, J. Food Prot. 71, pp. 1357–1365.
[5] Perni, S., Liu, D.W., Shama, G., Kong, M., 2008. Cold Atmospheric plasma decontamination of the
pericarps of fruit. Journal of Food Protection 71 (2), 302–308.

1644
 Intensification of process of water-thermal treatment of wheat grain before bread flour
milling
O. Safonova a, O. Razborskayab, V. Yuferov c, O. Ozerov d

a
Department of Foodstuffs Processing Technology, Petro Vasilenko Kharkiv National Technical
University of Agriculture, Kharkiv, Ukraine (avgust23@ukr.net)
b
Department of Foodstuffs Processing Technology, Petro Vasilenko Kharkiv National Technical
University of Agriculture, Kharkiv, Ukraine (neona84@ukr.net)
c
Institute of plasma electronics and new methods of acceleration, National Science Center “Kharkov
Institute of Physics and Technology” (v.yuferov@kipt.kharkov.ua)
d
Institute of plasma electronics and new methods of acceleration, National Science Center “Kharkov
Institute of Physics and Technology” (ozerov at @kipt.kharkov.ua)

INTRODUCTION
There is a new method of water-thermal treatment of wheat grain [1] in pneumopulsing device
by a wide range of acoustic waves at low pressure. This treatment can significantly reduce the
length of a given technological operation for several minutes. Application of this method is
more appropriate if wet processing of grain is used or for a small power plant, either for the
companies that are being built.
The aim of research is to examine the impact of the intensive WTT on technological properties
of wheat, which was processed by acoustic oscillations at low pressure before baking varietal
grinding.

MATERIALS & METHODS

The objects of investigated were wheat, flour, non yeast dough and yeast dough. Wheat was
characterized by the following quality: humidity - 12,7%, vitreousness - 50%, wet gluten
content - 18,0%. The grain of the wheat subjected to WTT in the following modes: 1 sample -
up to 15,5-16,0% humidity for 20 hours at a temperature of 18-20 oC (a traditional two-stage
cold WTT), 2 sample - were treated in pneumopulsing device pulsed power fluctuations - 3
units of device, pressure - 0,8 × 104 Pa, the number of oscillations - 120 impulses, 3 sample - 3
units of device, 2,4 × 104 Pa, 180 impulses, respectively.
The flour was produced from the grain at the laboratory mill MLU-202 Buhler. The hardness
of the grain was determined by using the experimental laboratory equipment. The degree of
swelling of the grain was measured by the change of its volume, flour whiteness was measured
with a device VBB-2m. To evaluate the strength of flour quantity and quality of gluten was
determined in the standard way, the sedimentation index of flour – a modified method of
Zeleny, to evaluate the physical properties of dough - farinograph Brabender, alveograph
Chopin. Amylolytic transformations were studied using amylograph and «Falling Number»
device.

RESULTS & DISCUSSION

Studying the changes of flour-milling properties of the grain after intensive WTT showed that
the effect of acoustic waves at low pressure on the hardness of the grain at axial compression is
reduced by 24,5%, the volume of the grain is increasing to 17,0-18,5%, yield of flour is rising
by 2,5-2,6%, and whiteness of flour - at 7-7,5 units. Unit energy consumption for grinding was

11th International Congress on Engineering and Food - Athens, Greece, 2011 1645
reduced to 22,5% in comparison to untreated grain. After intensive WTT the quantity of gluten
increased by 2,0-2,5%. To evaluate the strength of flour the sedimentation index of flour was
determined, which characterizes the quality of flour [2]. In this study the sedimentation index
of flour is the lowest in sample 1, which was prepared by the traditional cold WTT, and the
highest - in sample 2 (6,5%) which was subjected to impulsive treatment under less pressure.
Sample 3 has less sedimentation index compared to sample 2, but it is higher compared to
sample 1 (4,3%). Therefore, samples, subjected to impulsive processing, are characterized by a
higher sedimentation index, and thus the strength of flour is higher compared to the traditional
cold WTT. Studying the physical properties of non yeast dough in farinograph for 2 hours
showed that the flour subjected to traditional cold WTT (sample 1)is characterized by a higher
index of the rarefaction of the dough (220 units of farinograph (un. f.)) compared with the
experimental samples (170-210 un. f.). Samples processed by impulses (samples 2, 3) have
similar elasticity of dough, which exceeds the index of sample 1, on average, 10-20 un.f.
Similar changes are in the yeast dough: the samples processed by impulses are more elastic and
the index of the rarefaction is significantly lower (especially in sample 2) than in traditional
cold WTT. There is some improvement of baking properties of flour (strengthening of flour
and improving the physical characteristics of the dough) after intensive WTT of the wheat
grain in the pneumopulsing device before baking varietal grinding. Studying the physical
properties of the dough by using amylograph showed that after intensive WTT of the grain the
index of the elasticity of the flour increased to 12,7-14,3%, the strength of the flour increases
by 40-60 units of alveograph compared to the traditional cold WTT. Studying the amylolytic
properties of flour on amylograph shows that the maximum viscosity of the intensively treated
samples is higher by 30-40% compared to the traditional cold WTT. Higher indexes in these
samples were also got on «Falling Number» device. These facts indicate some amylolytic
transformations in the flour. Trial laboratory baking showed that intensive WTT of the grain
before a baking varietal grinding does not worsen the quality of the final products.

CONCLUSION
Studying the influence of the new WTT on the grain showed that under the conditions of
certain parameters of intensive treatment (power of the impulses, pressure and number of
impulses) the same technological properties of the grain can be achieved after the traditional
cold WTT. The duration of the WTT is considerably reduced and the grain is prepared for
grinding much quicker.

REFERENCES
[1] Utility patent 50 802. Ukraine. B02B1/00. Method of water-thermal treatment of the grain of the
wheat before baking varietal grinding. Safonova O. N., Razborska O. O., Domnich M. I., Yuferov V.,
Ozerov O. M., Ponomaryev O.N. Applicants and patentees: Safonova O. N., Razborska O. O. –
ʋ u 2009 13204; Publ. 12/18/2009 City; publ. 06/25/2010, Bull. Number 12.
[2] Vasilenko I. N. & Komarov V. I. 1987. Evaluation of grain quality: a reference. - Moscow:
Agropromyzdat, USSR.

1646
 The effect of abiotic stress pre-treatments on quality attributes of fresh-cut carrot cv.

Nantes

Carla Alegriaa,c, Joaquina Pinheiroa, Margarida Duthoita, Elsa M. Gonçalvesa, Maria Teresa Coelhob,
Margarida Moldão-Martinsc, Marta Abreua

a
UITA/INRB, Lisbon, Portugal (marta.abreu@inrb.pt)
b
Escola Superior Agrária de Castelo Branco, Castelo Branco, Portugal (mteresacoelho@ipcb.pt)
c
SCTA/DAIAT. ISA. Technical University of Lisbon, Lisbon, Portugal (mmoldao@isa.utl.pt )

INTRODUCTION
Abiotic stresses, such as heat shock and UV-C radiation, can be used to induce the synthesis of
bioactive compounds and prevent decay in fresh-cut fruits and vegetables. Among the
preservation techniques that are currently in use by the fresh-cut industry, e.g. antioxidants,
chlorines and modified atmosphere packaging, the use of heat shock and ultraviolet-C
treatments, alone or in different combinations have proved useful to control microbial growth
while maintaining quality during storage of fresh-cut produce [1].
The aim of this study was to evaluate the effects of heat shock and UV-C radiation stress
treatments, applied in whole carrots, on the overall quality of fresh-cut carrot cv. Nantes during
storage (5 ºC).

MATERIALS & METHODS

Raw whole carrots (Daucus carota L. cv. Nantes) without treatment (Ctr_samples) and
submitted to heat and UV pre-treatments (HS_ and UV_samples, respectively) were
subsequently minimal processed. Minimal processing (MP) included peeling, shredding,
decontamination process (chlorinated water, 200 ppm /1 min), rinsing, drying and packaging
operations. Heat shock was performed by immersion of whole peeled carrots in hot-water at
100 ºC for 45 s and paper dried. UV-C pre-treatment were prepared in a UV-C apparatus and
the whole peeled carrots were placed in a single layer for 2 min (0.78±0.36 kJ.m-2). After HS
and UV-C pre-treatments, the whole carrots were held at 5 ºC for 24h until minimal processing
operations without the decontamination step. Analytical procedures were carried out in
triplicate on five sampling dates, i.e. days 0, 3, 5, 7 and 10. Total mesophilic aerobic count
(TAPC) [2], headspace gas (O2/CO2, %) analysis (Abissprint), total phenolic content [3], total
carotenoid [4], peroxidase (POD) [5] were determined and colour whitening index (WI) was

calculated by the expression:


WI 100  100  L * 2  a * 2 b * 2. Statistically significant
differences (P<0.05) between samples were determined according to LSD test (ANOVA).

RESULTS & DISCUSSION

Heat shock (HS, 100 ºC/45s) and UV (0.78r0.36 kJ.m-2) samples had higher phenolic content
and exhibit reduced POD activities during storage regarding Ctr samples (200 ppm free
chlorine/1 min). All analyzed samples showed reduced carotenoid contents considering raw

11th International Congress on Engineering and Food - Athens, Greece, 2011 1647
material (unprocessed carrot). Nonetheless, UV samples registered a three-fold increase in
carotenoid content in subsequent storage. Colour of fresh-cut carrot showed a continuous
increase in WI values during storage regardless of treatment without impairing quality
acceptance. Respiratory metabolism was affected by both abiotic stress treatments since
reduced O2/CO2 rates were found, more significant in HS samples. The decontamination effect
of the tested stresses was more expressive in HS samples, where a § 2 Log10cfu.g-1 reduction in
initial microbial load was achieved and also provided reduced microbial development rate
during storage. Both stress treatments show beneficial effects on the overall quality of fresh-cut
carrot and proved to be more efficient than the industrial practice for fresh-cut carrot.

CONCLUSION
Both stress pre-treatments showed beneficial effects on the quality of fresh-cut shredded carrot,
namely increases in phenolic and carotenoid contents and also reduced POD activity during
storage. Moreover, heat shock pre-treatment promotes an effective decontamination effect and
reduced respiratory levels.

REFERENCES
[1] Allende A., Tomás-Barberán F.A. & Gil M.I. 2006. Minimal processing for healthy traditional foods.
Trends in Food Science & Technology, 17, 513-19.
[2] EN ISO 4833: 2003. “Microbiology of food and animal feeding stuffs - Horizontal method for the
enumeration of microorganisms - Colony-count technique at 30ºC”.
[3] Singleton V.L. & Rossi J.A. 1965. Colorimetry of total phenolics with phosphomolybdic-
phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144-158.
[4] Biehler E., Mayer F., Hoffman L., Krause E. & Bohn T. 2010. Comparision of 3 Spectrophotometric
Methods for Carotenoid Determination in Frequently Consumed Fruits and Vegetables. Journal of
Food Science, 75(1), C55-C61.

1648
Yogurt from ultrasound treated milk: monitoring of fermentation process and evaluation

of product quality characteristics.

Panagiotis Sfakianakisa & Constantina Tziab

Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical
University of Athens, 5 Iroon Polytechnioy St.,15780 Athens, Greece
a
psfakian@central.ntua.gr, btzia@chemeng.ntua.gr

INTRODUCTION
Milk and milk products, in order to become safe and palatable for consumption, undergo a
series of processes in the dairy industry, i.e. pasteurization, standardization (SNFC and fat
content) and homogenization. Homogenization is an important step that aims at reduction of
the milk fat globules size from about 10-12 ȝm to 1-2 ȝm and therefore at prevention of their
separation from the main volume of the milk. The principle of homogenization is the
application of extreme conditions to the milk fat globules, so they disperse and reform into
smaller. The most commonly used method in dairy industry is pressure homogenization.[3] An
alternative method efficient for milk homogenization is ultrasound treatment. High intensity
ultrasound causes vibrations and cavitation phenomena and thus the membrane that surrounds
the milk fat globules dissipates and reforms creating smaller globules [1]. Several studies have
applied ultrasound in milk homogenization and have shown that the milk fat globules (MFG)
reduce drastically in size the more intensive the ultrasound treatment in. Furthermore the use of
ultrasound treated milk has effect on yoghurt production and on yoghurt itself. The
fermentation process was longer, but the final product had superior rheological properties,
strong gel structure, high water holding capacity and low syneresis [4]. The present study will
apply ultrasound on raw bovine milk and afterwards, using the ultrasound homogenized milk,
produce yogurt.

MATERIALS & METHODS

Raw bovine milk (3,5% fat, 3,3% SNFC), after being heated at 60 oC, was treated by
ultrasound for 10min at amplitudes 150, 267,5, 375, 562,5 and 750 W, then pasteurized at
85 oC for 20min, cooled at 46 oC and inoculated with the started culture (industrial symbiotic
culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. Bulgaricus) and
incubated in 45 oC until the pH value of 4,7. After that the samples were stored to 4 oC for 24h.
After the ultrasound treatment MFG were pictured at an optical microscope. During
fermentation pH was monitored as well as viscosity. At the final product, viscosity and texture
properties, hardness, adhesiveness, cohesiveness and gumminess, were measured.

RESULTS & DISCUSSION

Ultrasound homogenization led to decrease of MFG size. Low amplitude (150W) had not a
satisfactory homogenization effect; the MFG size and distribution were similar to untreated
milk. Medium amplitude ultrasound (267.5, 375 W) had a good homogenization effect; MFG
average diameter was 2ȝm. Higher amplitude ultrasound reduced the MFG size a lot, making

11th International Congress on Engineering and Food - Athens, Greece, 2011 1649
them barely visible at the optical microscope (100x magnification); their average diameter size
was 0,3ȝm. During fermentation process the pH decrease rate of each milk sample,
homogenized by ultrasound, was slower compared with the respective described in the
literature. Instead of 3 to 4h, the fermentation of ultrasound treated milk lasted 4-5h. However
during fermentation the viscosity increase was rapid and the final viscosity values were high,
ranged from 1,4 to2 Pas*s for medium and high amplitude ultrasound (375, 562.5, 750 W) of
milk treatment. Both pH decrease and viscosity evolution for all samples followed the
sigmoidal course described by the modified Gompertz model [4]. The figure presents the pH
and viscosity change during fermentation of a milk sample homogenized by 375W ultrasound.

Figure 1. pH decrease and Viscosity evolution during fermentation of milk sample homogenized
by 375 W ultrasound.

The yoghurt produced from ultrasound homogenized milk had strong coagulum presenting
high viscosity values, ranged from 1-2 Pa*s. Also the texture characteristics (hardness
cohesiveness, adhesiveness and gumminess) were improved, The higher the intensity of the
ultrasound treatment, the higher viscosity and improved texture characteristics values were.

CONCLUSION
Concluding, high intensity ultrasound had a good homogenization effect, but leads to longer
fermentation process duration. The yogurt produced from ultrasound treated milk had higher
viscosity, stronger coagulum and superior texture characteristics.

REFERENCES
[1] Z.J. Dolatowski, J. Stadnik, D. Stasiak. Applications of Utrasound in Food Technology, Acta Sci.
Pol. Technol. Alimemnt. 2007, 6(3): 89-99
[2] H. Wu, G.J.Hulbers, J.R.Mount. Effect of Ultrasound on Milk Homogenization and Fermentation
with Yoghurt Starter, Innovative Food Science & Emerging Technologies, 2001, 1: 211-218
[3] P. Walstra, J.T. WWouters, T.J. Geurts. Dairy Science and Technology, CRC Press Taylor &
Francis Group 2006
[4] C. Soukoulis, P. Panagiotidis, R. Koureli, and C. Tzia. Industrial yogurt manufacture: Monitoring of
fermentation process and improvement of final product quality, J. of Dairy Science, 2007, 90:
2641-2654.

1650
 Effect of Sonication on Malting Behaviour of Barley

Emmanuel Dutheila, Brijesh Tiwarib, Mahesh Guptac, PJ Cullend, Charles Brennanb, Colm O'Donnella

a
Biosystems Engineering, University College Dublin, Dublin, Ireland
b
Department of Food, Manchester Metropolitan University, Hollings Faculty, Manchester, UK
c
Food and Environmental Health, Dublin Institute of Technology, Dublin, Ireland

INTRODUCTION
Germination capacity of barley grains plays a significant role in the brewing process of barley.
Germination capacity of barley grain is influenced by both pre-harvest and post harvest stages
[1]. Steeping time and temperature is one of the critical process influencing the modification
of endosperm materials of barley malt [2]. Several researchers examined various techniques to
reduce water usage during steeping for improving process efficiency and cost. Power
ultrasound as a novel processing method finds wide application in various food processing
operations including fruit juice preservation [3], enhancing drying rate [4] and extraction of
bioactive compounds [5]. The objective of this study was to investigate the effect of sonication
on the malting behaviour of barley grains.

MATERIALS & METHODS

Malting barley cultivar grains were obtained from a brewery (Guiness, Dublin, Ireland), sieved
and cleaned. One kg of cleaned barley grains were sonicated in 3 L of distilled water using an
ultrasonic bath (Bransonic® 5210E DTH). Sonication treatment was performed for 20, 40 and
60 min at a constant frequency of 47 kHz ± 6% at a set temperature of 25 oC. Sonicated and
control barley grains were steeped for 24 h by immersion in water at 16 °C and subsequently
germinated for 96 h. Samples were kilned at 50 °C for 16 h before manually derooted to obtain
malt. Pasting properties of sonicated barley grain and control samples obtained after milling
samples to flour were determined using a Rapid Visco Analyser. The tetrazolium chloride (TZ)
test was employed to determine seed potential viability and vigour. Determination of the ȕ-
glucan content of the two isolates was carried using a Megazyme® mixed linkage ȕ-glucan
assay.

RESULTS & DISCUSSION

The effect of sonication on acrospire length (mm) during germination is shown in Figure 1. A
significant increase in the length was observed with an increase in sonication time. The length
of acrospire was higher compared to control at any given sonication treatment during a
germination period of 96 h, clearly demonstrating that sonication enhances the germination
rate. The TZ test also indicated a 100 % grain germination rate for samples sonicated for 60
min compared to a rate of 95 % for samples sonicated for 20 and 40 min and an 80% rate for
control samples. Germination tests showed that sonication enhances both seed viability and
vigour. The sonication effect on germination is mainly due to sonochemical reactions involving
physical i.e. acoustic cavitations and chemical reactions [5]. The cavitations during sonication
may also induce microfissures on the grain surface and improve imbibition of moisture for
enhanced germination rate. The RVA profile of sonicated barley grains is shown an increase in

11th International Congress on Engineering and Food - Athens, Greece, 2011 1651
peak viscosity over control was observed at 20 and 40 mins, PV was found to decrease at 60
min treatment time. A significant increase in final viscosity was observed in sonicated samples.
The inconsistent changes in pasting profile of sonicated grains might also be influenced by
thermal effect induced by sonication. An increase in the temperature during the sonication from
25 °C to 33 °C for 20 min, 35.8 °C for 40 min and 43.9 °C for 60 min was observed.

50
Lengthofacrospire (mm)

40

30

20

10

0
0 24 48 72 96
Germinationtime(h)
Figure 1. Changes in length of acrospire during germination

No significant difference was observed for the ȕ-Glucan content of both sonicated (0.55 – 0.86
mg/100gm) and control (0.97mg/100g) samples. However a significant decrease was observed
during malting. A significant decrease of about 42.9 – 46.7% for sonicated and about 39.9 %
for control samples was observed during malting. A decrease in ȕ-glucan content during
sonication process and subsequent germination might be due to several reasons including
solublisation of soluble ȕ-glucan during sonication process and enzymatic degradation of ȕ-
glucan due to ȕ-glucanase. It has been reported that low power sonication enhances enzymatic
activity.

CONCLUSION
This study shows that sonication improves the germination capacity of barley grains.
Ultrasound as a pre-treatment during barley steeping process can help in improving
germination by reducing germination time and improving process efficiency.

REFERENCES
[1] Tiwari, U & Cummins, E., (2009). Factors influencing ȕ-glucan levels and molecular weight in cereal-based
products, Cereal Chemistry, 86, 290–301. [2] Bryce, J. H., Goodfellow, V., Agu, R. C., Brosnan, J. M., Bringhurst, T.
A., & Jack, F. R. (2010). Effect of Different Steeping Conditions on Endosperm Modification and Quality of Distilling
Malt. Journal of the Institute of Brewing, 116(2), 125-133. [3] Goussous, S. J., Samarah, N. H., Alqudah, A. M., &
Othman, M. O. (2010). Enhancing seed germination of four crop species using an ultrasonic technique. Experimental
Agriculture, 46(02), 231-242. [4] O'Donnell, C. P., Tiwari, B. K., Bourke, P., & Cullen, P. J. (2010). Effect of
ultrasonic processing on food enzymes of industrial importance. Trends in Food Science & Technology, 21(7), 358-
367. [5] Mason, T. J., Chemat, F., & Vinatoru, M. (2011). The Extraction of Natural Products using Ultrasound or
Microwaves. Current Organic Chemistry, 15(2), 237-247.

1652
 A mathematical approach for using multiple enzyme based pressure-temperature-time

integrators (PTTIs) for high pressure process evaluation

Eleni Gogou, Petros Taoukis

School of Chemical Engineering, National Technical University of Athens, Greece

(egogou@chemeng.ntua.gr)

INTRODUCTION
In order to ensure the optimization and control of high pressure processing, evaluation of the
process impact on both safety and quality attributes of foods is essential. Enzymes can serve as
effective tools in evaluating the impact of high pressure processes of foods [1, 2]. Pressure-
Temperature Time Integrators (PTTIs) show in a single response (i.e. the measurement of
activity change after HP treatment) the integral effect of process temperature and pressure (i.e
F-value) on the target index (a microorganism or an enzyme of the processed food). The
objective of this study was to investigate the combined use of enzyme systems as PTTIs in
quantifying the F-value of typical HP processes of food products.

MATERIALS & METHODS

The effect of high pressure processing on 80 800
the activity of selected enzymes was
(3)
formerly studied [2, 3, 4]. High pressure
600
processing was performed under several
Temperature (°C)

60 (2)

Pressure (MPa)
isobaric conditions (100, 200, 300, 450 (1)
and 600 MPa) combined with temperature 400
40
ranging from 25 to 70 °C. The
ȆȓİıȘ (MPa)
inactivation kinetics of enzymes such as 200
Ĭ

ĬİȡȝȠțȡĮıȓĮ
xylanases, tyrosinase, lipase and 20 ȆȓİıȘ
pectinmethylesterases were used in
0
simulated pressure-temperature-time 0 1 2 3 4 5
ȓ (°C)

profiles typical of HP processing (Figure Time

ȋȡȩȞȠȢ(min)
(min )
1). These simulations were used in order
to investigate the use of PTTIs for the Figure 1. Typical simulated pressure-temperature
evaluation of HP processes. profiles of HP processes.

RESULTS & DISCUSSION

The application of the enzyme based PTTIs was evaluated in simulated HP processes of
different target indices at different pressure- temperature conditions designed to achieve the
targeted F-values. The effectiveness of each of these processes could be evaluated by the PTTI
response i.e. the value of enzyme remaining activity. The PTTI responses that would be
obtained for the HP processes were calculated from the integration of the developed kinetic

11th International Congress on Engineering and Food - Athens, Greece, 2011 1653
model of enzymes inactivation. The enzyme inactivation of enzymatic PTTIs was translated to
F-values and compared to the F-value of the processes for different target indices.
Single PTTI response allowed reliable calculation of the process F-value process when both the
values of activation energy, Ea, and activation volume, Va, of PTTI and target index are in the
same range and show similar dependency on pressure and temperature respectively. This is a
very strict requirement.
700
Alternatively a double/multiple
(ǻ3) PTTI system can be used. The
600 responses after a certain process of
a double PTTI system (based on
500
ȆȓİıȘ (MPa)

two different enzymes e.g. a

400 xylanase and a tyrosinase) can be
Pressure

translated to F-values using the

300 PTTI kinetics and corresponded to
200 distinct isorate contour lines
showing equivalent combinations
100 of pressure and temperature
20 30 40 50 60 70 80 90
(Figure 2). The contours’ intercept
ĬİȡȝȠțȡĮıȓĮ (°C)
Temperature (°C) (shown in Figure 2) defines the
Figure 2. Contour plots of pressure and temperature process effective pressure and
combinations resulting in the same equivalent HP process temperature. These effective
impact (Fvalue) for four different enzyme based PTTIs process conditions are used for
(tyrosinase, xylanase, lipase, pectinmethylesterase). calculation of the actual F-value of
the HP process. The results for a
large number of simulated HP pressure-temperature-time profiles, showed difference of less
than 18% between the PTTI calculated F-value based on double or triple enzyme systems, and
the F value of the food.
CONCLUSION
It was demonstrated that the combined use of multiple enzyme PTTIs allows a satisfactory
mathematical evaluation of HP processes for different target indices.
REFERENCES
[1] Van der Plancken I, Grauwet T, Oey I, Van Loey A, Hendrickx M. 2008. Impact evaluation of high
pressure treatment on foods: considerations on the development of pressure temperature-time
integrators (pTTIs). Trends in Food Science & Technology 19(6):337-348.
[2] Gogou E., Katapodis P., Taoukis P.S. (2010). High Pressure Inactivation Kinetics of a Thermomyces
lanuginosus Xylanase Evaluated as a Process Indicator. Journal of Food Science, 75: E379–E386.
[3] Gogou E. & Taoukis P.S. (2009). Kinetic study of the inactivation of lipase under high hydrostatic
pressure treatment and the development of a pressure temperature time integrator (PTTI). Institute of
Food Technologists (IFT) Annual Meeting, Anaheim, OC CA, USA, June 06-09, 2009. Book of
abstracts p.215.
[4] Gogou E., Katsaros G., Boulekou S. & Taoukis P.S. (2008). Use of enzymes of different inactivation
kinetic characteristics as high pressure temperature time integrators (PTTIs). Institute of Food
Technologists (IFT) Annual Meeting, N. Orleans, Louisiana, USA June 28- July 01, 2008. Book of
abstracts p.182.

1654
 Effect of high hydrostatic pressure treatments on physicochemical properties, microbial

quality and sensory attributes of beef carpaccio

N. Szermana,b, Y. Barriob,c, B. Schroederc, P. Martinezc, A. Sanchoa, C. Sanowa, S.R. Vaudagnaa,b,c,d

a
Instituto Tecnología de Alimentos, CIA, INTA,CC77, Morón CP:B1708WAB, Argentina
b
CONICET, Buenos Aires, Argentina (e-mail: yanina.barrio@conicet.gov.ar)
c
Facultad de Ingeniería y Ciencias Exactas, UADE, Buenos Aires, Argentina
d
Facultad de Agronomía y Ciencias Agroalimentarias, Universidad de Morón, Morón, Argentina

INTRODUCTION
High hydrostatic pressure (HHP) technology has been successfully applied for the processing of cured
meat products -cooked or dried- and cooked ready-to-eat meats. In the case of ready-to-eat cured fresh
meat (i.e. carpaccio), the HHP technology can be an alternative for product pasteurization, assuring food
safety and extending shelf-life [1]. However, the application of HHP on fresh pigmented meats causes an
important discoloration, particularly at pressure levels above 300MPa, which are required for vegetative
cells inactivation [2]. HHP at refrigeration temperatures applied to previously frozen samples could
improve the appearance of beef carpaccio in comparison to HHP treatments at moderate (room)
temperature. The aim of this study was to evaluate the effect of sample conditioning (frozen or thawed)
and HHP treatments (different pressure levels at refrigeration or moderate temperature) on
physicochemical properties, microbial quality and sensory attributes of beef carpaccio.

MATERIALS & METHODS

For each carpaccio conditioning (frozen or thawed), a factorial randomized block (2) design was applied
with temperature (two levels: 0°C and 5ºC -frozen samples-; 5°C and 20ºC -thawed samples-) and
pressure (four levels: 0, 400, 500, 600MPa) as main factors. Holding time at working pressure was 5 min.
Control samples were fresh carpaccio non-submitted to HHP treatments (0MPa). Semitendinosus beef
muscles (21) were intermittently tumbled for 60min (5rpm-2min on / 8min off) with sodium chloride
(12g/kg), sodium tripolyphosphate (1g/kg), sodium citrate (0.5g/kg), sodium nitrite (0.15g/kg) and
sodium isoascorbate (0.5g/kg). Then, muscles were packed under vacuum and stored at 1ºC for 12 days.
After chill storage, cured muscles were frozen (-40ºC), sliced (slice thickness: 1.5-2mm), packed under
vacuum and stored at -40ºC. Frozen or thawed (4°C) samples were submitted to different HHP treatments
according to the experimental design. After HHP treatment, samples were stored at -40°C until further
testing. Analysis performed in all samples (previously thawed) were: expressible moisture (EM) by
centrifugation method, pH on meat slurries, shear force (SF) and work of shearing (WS) measured using a
10 blade Kramer shear cell, chromatic parameters (CIELab), sensory appearance (triangular test), and
aerobic total count (ATC) at 30ºC.

RESULTS & DISCUSSION

Frozen or thawed carpaccio treated by HHP presented EM values (Table 1) significantly higher (p<0.05)
than their respective controls. SF values of frozen or thawed samples treated by HHP (Table 1) were
higher than control ones, but those differences were not significant (p>0.05). Regarding WS, all frozen
samples treated by HHP and thawed ones treated at 600MPa presented significantly higher (p<0.05)

11th International Congress on Engineering and Food - Athens, Greece, 2011 1655
values than controls. A significant increase (p<0.05) of pH (Table 1) was observed in thawed samples
treated by HHP compared to their control sample. L* parameter values significantly (p<0.05) increased in
all samples treated by HHP (frozen or thawed) in relation to their control, probably caused by globin
denaturation and/or to heme displacement at pressures above 300MPa [2]. For all samples treated by
HHP, a* parameter values were lower than control ones, and b* parameter was significantly (p<0.05)
reduced in thawed samples. Sensory appearance of all treated samples was significantly different (p<0.01)
from control one. Maximum and minimum ATC reductions were 5.24log10 cycles at 600MPa-5ºC and
3.79log10 cycles at 400MPa-5ºC for thawed samples treated by HHP and 4.11log10 cycles at 500MPa-0ºC
and 2.74log10 cycles at 400MPa-0ºC for frozen ones.

Table 1. Expressible moisture (EM), shear force (SF), work of shearing (WS), pH and chromatic
parameters of frozen or thawed carpaccio treated by HHP at different pressures and temperatures.
T P EM SF WS Chromatic Parameters
pH
(°C) (MPa) (%) (N) (Nm) L* a* b*
0 400 26.56±3.09* 232.53±52.56 0.66±0.17* 5.87±0.12 48.04±0.82* 26.76±2.06* 18.65±1.03
0 500 28.33±3.46* 239.00±54.48 0.69±0.21* 5.88±0.08 47.72±2.37* 27.58±2.83* 19.03±1.93
Frozen

0 600 28.83±1.95* 223.32±30.48 0.66±0.12* 5.88±0.11 46.49±3.03* 27.64±1.61* 18.14±0.79

5 400 29.71±1.96* 247.82±55.57 0.73±0.17* 5.90±0.19 48.21±1.92* 25.88±1.45* 17.61±0.91
5 500 28.46±3.08* 235.53±57.17 0.64±0.15* 5.84±0.05 49.05±2.23* 26.46±2.72* 18.56±0.71
5 600 28.73±3.79* 241.48±25.94 0.69±0,09* 5.87±0.13 48.49±2.54* 26.26±1.78* 18.16±1.33
Control 20.49±4.18 199.05±18.78 0.53±0.04 5.79±0.07 44.50±2.74 29.77±2.24 18.86±1.16
5 400 25.57±2.62* 215.78±35.31 0.59±0.08 6.01±0.12* 57.13±3.50* 22.59±2.75* 15.37±1.22*
5 500 26.25±2.55* 220.82±59.43 0.59±0.18 5.92±0.07* 57.26±2.43* 22.82±2.20* 16.21±1.21*
Thawed

5 600 26.34±1.89* 234.78±35.17 0.65±0.12* 5.97±0.08* 56.56±3.13* 22.59±1.61* 16.38±1.33*

20 400 25.96±1.82* 241.85±45.76 0.66±0.12 5.98±0.11* 58.22±0.98* 21.92±0.53* 15.50±0.88*
20 500 26.29±2.34* 222.58±65.09 0.63±0.24 5.98±0.06* 55.91±1.17* 23.07±2.38* 14.53±1.79*
20 600 27.10±1.67* 236.32±56.13 0.69±0.20* 5.97±0.07* 56.88±3.42* 22.64±2.84* 15.65±0.76*
Control 22.33±3.99 190.78±38.42 0.51±0.08 5.88±0.07 45.33±3.35 29.89±3.18 19.77±0.80

CONCLUSION
Temperature applied during HHP processing of thawed (5 and 20°C) or frozen (0 and 5°C) carpaccio did
not have a significant effect on the measured parameters. Pressure level effect was only observed on ATC
and work of shearing of thawed treated samples. Frozen conditioning of carpaccio previous to HHP
treatments reduced the harmful effects of pressure on chromatic parameters (L*, a* and b*) and water
holding capacity. It was observed a lower effectiveness of HHP treatments on microorganisms’
inactivation for frozen carpaccio than for thawed one, probably as a result of the lower water activity of
the former.

REFERENCES
[1] Fernández P.P., Sanz P.D., Molina-García A.D., Otero L., Guignon B. & Vaudagna S.R. 2007. Conventional
freezing plus high pressure-low temperature treatment: physical properties, microbial quality and storage stability
of beef meat. Meat Science, 77(4), 616-625.
[2] Carlez, A., Veciana-Nogues, T., Cheftel, J.(1995). Changes in colour and myoglobin of minced beef meat due to
high pressure processing. Lebensmittel-Wissenschaft und-Technologie, 28(5), 528-538.

1656
 Rheological properties of high pressure milk cream

Donsì G.a,b Ferrari G.a,b, Maresca P.b

a
Department of Industrial Engineering, University of Salerno,
via ponte don melillo, 84084 Fisciano (SA), Italy
b
ProdAl Scarl, University of Salerno,
via ponte don melillo, 84084 Fisciano (SA), Italy (info@prodalricerche.it)

INTRODUCTION
Several studies reported in the literature demonstrated the pressure-induced modifications of
the main milk constituents [1,2,3]. The observed denaturation of milk macromolecules depends
on the applied pressure level, treatment time and temperature level. As a result of the protein
denaturation and micelles solubilisation, the proteins conglomerate with milk fat globules,
influencing the viscosity, viscoelasticity and creaminess and, in particular cases, causing the
gelation of the HP treated milk and allowing the application of HP milk in innovative dairy
productions. The aim of this paper was to setup the process cycles to produce a milk cream
under pressure. Several experiments were carried out to evaluate and model its rheological
behaviour. A preliminary concentration of the milk was carried out with the aim to enhance the
creaminess of the high pressure milk cream. Finally the experimental data attained were
analyzed in order to characterize the rheological behaviour of the HP milk cream.

MATERIALS & METHODS

Pasteurized and homogenized whole milk, kept on the local market, was concentrated (50%,
60%, 70%) by means of a rotary evaporator. The samples were stored under refrigerated
conditions until the High Pressure(HP) treatments carried out in a pilot plant MINI FOODLAB
FPG5620 (SFP Ltd, UK). Samples were preliminary treated at variable processing conditions
(pressure= 400-500 MPa, time= 5-10 min, temperature = 25 °C) to setup the process cycle.
Samples were, then, processed under the optimized processing conditions and the rheological
behaviour analyzed. Shear flow and stress sweep measurements were carried out on the fresh,
pre-concentrated and HP treated milk samples.

RESULTS & DISCUSSION

The experimental campaign was aimed to choose the optimal formulation and the process
conditions (pressure-temperature-time levels) to obtain a product with the creaminess and the
texture of a dairy cream. Firstly to setup the process cycle, the HP experiments were carried out
on pre-concentrated milk samples (50%) and the oscillatory rheological tests performed on the
fresh, concentrated and HP treated samples. The milk coagulation occurs applying a minimum
pressure level of 500 MPa and operating time of 10 min. Three milk formulations (50%, 60%;
70% pre-concentration), were treated under the optimized conditions and the rheological
behaviour analyzed. The results of the stress sweep measurements are shown in Figure 1. The
fresh and pre-concentrated milk show a prevalent viscous behaviour, being the values of the
elastic modulus round down to zero. The milk pre-concentration increases the values of the
viscous modulus without modifying the rheological behaviour of the samples.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1657
 100 100
50%HP 60%HP
90 90
70%HP yogurth0.1%
80 80
yogurth3,7% yogurth4,1%
70 70

60 60

G"(Pa)
G'(Pa)

50 50

40 40

30 30

20 20

10 10

0 0
0 2 4 6 8 10 12 14 0 2 4 6 8 10 12 14
strain(Pa) strain(Pa)
Figure 1 Elastic (G') and viscous (G") moduli measured for pre-concentrated (50%, 60%, 70%) high
pressure treated (HP) milk samples and commercial yogurts with different fat content (0.1 %, 3.7 %, 4.1 %)
in stress sweep tests (Pressure:500 MPa, temperature: 25°C, processing time: 10 min).

The HP treated samples, indeed, clearly show an evident viscoelastic behaviour. In this study
the elastic and the viscous moduli were used to estimate the creaminess of the pressure-induced
cream and the yogurt samples. In the case of yogurt samples, a higher creaminess perception is
associated with a higher fat content. The trends observed for the 50%-HP and 60%-HP dairy
cream overlap with the ones of the light and full-fat yogurt respectively. The 70%-HP dairy
cream shows higher values of the G’ and G” parameters, by far exceeding the values measured
for the other analyzed samples and demonstrating a higher compactness. HP samples may
ensure a creaminess and a texture perception similar to the commercial dairy products.
Moreover the fresh milk shows a typical Newtonian behaviour. Even if the same model holds
for the 50% and 60 % pre-concentrated milk, the more concentrated milk as well as the HP
treated samples shows a typical shear thinning behaviour, decreasing the apparent viscosity
upon increasing the shear rate. This result demonstrates that the pressure-induced dairy cream
is stabilized by weak interactions, which can be destroyed under stress conditions.

CONCLUSION
The experiments demonstrate that the milk coagulation occurs at a minimum pressure level of
500 MPa and a processing time of 10 min at ambient temperature. The milk pre-concentration
allows the rheological properties of the pressure-induced cream to be modified and improved.
The fresh milk and pre-concentrated milk show a prevalent viscous behaviour, while the
percentage of pre-concentration only increases the values of the viscous modulus. The high
pressure samples indeed, show an evident viscoelastic behaviour, due to the macromolecules,
which conglomerate under pressure and tend to align and provide greater resistance to flow.

REFERENCES
[1] Anema S. G., Lowe E. K. & Stockmann R. 2005. Particle size changes and casein solubilisation in
high-pressure-treated skim milk. Food Hydrocolloids, 19, 257í267.
[2] Hinrichs J. & Rademacher B. 2005. Kinetics of combined thermal and pressure induced whey protein
denaturation in bovine skim milk. International Dairy Journal, 15, 315í323.
[3] Huppertz T., Fox P. F. & Kelly A. L. 2004. High pressure treatment of bovine milk: Effects on casein
micelles and whey proteins. Journal of Dairy Research, 71, 97í106.

1658
 Effects of HHP combined with blanching on microorganisms and qualities of cloudy and clear

strawberry juices

Xiamin Caoa, Yan Zhanga, Xiaojun Liaoa, Xiaosong Hua

a
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China
(Liaoxjun@hotmail.com)

INTRODUCTION
The qualities of HHP-processed fruits and vegetables products changed during storage due to coexisting
chemical reactions when endogenous enzymes are incompletely inactivated. thus it was dispensable for
HHP-processed products to be refrigerated which increases the production cost and does not benefit the
commercialization. The purpose of this work was to evaluate the effects of HHP combined with
blanching on microorganisms and qualities of strawberry cloudy and clear juices, and to provide support
for designing the parameters of HHP commercialization in strawberry juices.

MATERIALS & METHODS

Figure 1. Flow chart of cloudy and clear juice preparation

To achieve microbiological safety, the parameters (600 MPa for 4 min) of HHP were finally selected for
qualities analysis.

RESULTS & DISCUSSION

The viscosity of HHP-treated cloudy juice was decreased by 12.40% (Fig. 2). In this study, blanching
prior to HHP inactivated PME, the viscosity of cloudy juice was not affected by inactivated PME. The
interactions between pectin molecules and other components in cloudy juice were produced by no-
covalent bonds including hydrophobic bonding, salt bridges, metalic bonds and intra molecular forces.
HHP can destroy these interactions. As shown in Fig 3, smaller viscosity corresponding to higher cloud in
HHP-treated cloudy juice and bigger viscosity corresponding to lower cloud in untreated cloudy juice was
manifested. On one hand, the gel formation of pectin catalyzed by PME in untreated cloudy juice caused
higher viscosity, and the pectin gel was more easily removed by centrifugation and thus lower cloud was
detected. On the other hand, the disruption of no-covalent interactions between pectin molecules and
other components in cloudy juice after HHP made the juice more stable during centrifugation and lead to
higher cloud.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1659
 0.35 8
7
A660; Viscosity
0.30 7

6 0.25 6

Viscosity (mPa.s)

Viscosity mPa S-1

0.20 5
5

A660
0.15 4

0.10 3
2

0.05 2

0.00 1
0 10 20 30 40 50 60 70
l HHP
contro HHP contro
l
juice
Shear rate (1/s) juice juice juice clear
cloudy cloudy clear

Figure 2. The flow behavior of cloudy and clear juice. Figure 3. Effect of HHP at 600 MPa/4 min/ambient
Ŷ untreated cloudy juice; Ƒ HHP cloudy juice; temperature on the cloud and viscosity of
Ÿuntreated clear juice; ƸHHP clear juice cloudy and clear juices.

Table 1. Effect of HHP on antioxidantive compounds and antioxidant capacity of cloudy and clear juices.
Ascorbic Anthocyanins Total Antioxidant capacity
Juices
acid Cy-3-glu Pg-3-glu Pg-3-rut phenols DPPH FRAP
Cloudy control 11.26±0.54 1.75±0.03 7.61±0.19 3.78±0.18 112.07±2.38 74.31±1.58 38.28±0.29
juice HHP 10.38±0.68 1.75±0.02 7.53±0.02 3.55±0.03 116.31±3.42 77.11±2.37 38.45±0.09
Clear control 6.43±0.23 1.44±0.01 7.38±0.11 2.39±0.03 86.45±2.18 50.34±1.22 26.31±0.22
juice HHP 5.62±0.18 1.42±0.01 7.37±0.04 2.34±0.04 89.79±1.73 49.89±3.28 25.85±0.07
Ascorbic acid and anthocyanins: mg/100 mL.Total phenols : equal to mg GAE/100 mL. DPPH and FRAP: equal to mg Vc/100 mL

In this study, there was 7.82% and 12.60% loss of ascorbic acid in HHP-treated cloudy and clear juices,
respectively. Monomeric anthocyanins exhibited no changes. HHP resulted in an increase trend of total
phenols in cloudy juice, which could be related to an increased solubilization of some antioxidant
components such as anthocyanins, amino acids and protein with phenolic hydroxyl group in cells into the
juice. No significant change of the antioxidant capacities in cloudy and clear juices was detected after
HHP.
Table 2. The effect of HHP on color of cloudy and clear juices
Color
Juices
* *
L a b* ǻE
Cloudy control 28.91f0.21 34.89f0.49 35.12f0.99 1.69
juice HHP 29.81f0.84 33.46f0.5 35.14f0.48
Clear control 47.36f1.22 37.96f0.83 50.23f0.88 3.64
juice HHP 50.57f1.39 36.25f0.98 50.63f1.05

The L* value of HHP-treated cloudy juice exhibited an increase tendency and of HHP-treated clear juice a
significant increase. The a* and b* value of the HHP-treated juices exhibited no significant change. The
ǻE value of cloudy juice was lower than 2 while that of clear juice was greater than 2, suggesting that
there was a visible color difference in clear juice after HHP.

CONCLUSION
HHP treatment (600 MPa/4 min/ambient temperature) combined with blanching could totally inactivate
natural microorganisms and well preserved the quality characteristics including cloud, anthocyanins and
total phenols, antioxidant capacity and color, and it was an effective preservation method of strawberry
juices.

1660
 Effect of high pressure homogenization process on Bacillus stearothermophilus and

Clostridium sporogenes spores in skim milk

Cláudia R. G. PINHO; Mark A. FRANCHI; Alline A. L. TRIBST; Marcelo CRISTIANINIa

a
Department of Food Technology (DTA), School of Food Engineering (FEA), University of Campinas
(UNICAMP), Campinas, SP, Brazil (olecram@fea.unicamp.br)

INTRODUCTION
High pressure homogenization (HPH) is an alternative food processing technique. It preserves
sensory and nutritional food characteristics due to low heating, being interesting to guarantee
the safety of thermo-labile food [1]. Milk is a food that can be spoilage by many bacteria
genera. The inactivation of sporulated microorganisms as Bacilli and Clostridia, in milk is a
challenge, since they are thermoresistant and important for milk deterioration. The present
work aimed to evaluate the inactivation of B. stearothermophilus ATCC 7953 and C.
sporogenes PA 3679 spores in skim milk by HPH.

MATERIALS & METHODS

For the tests, commercial UHT skin milk was inoculated with 105 spores.mL-1 and then
subjected to homogenization at pressures of 100, 200 and 300 MPa in a high pressure
homogenizer Stansted, model FPG 7400H:350 . The heat resistance of spores at 110, 115 and
121ºC were measure before and after HPH at 300MPa using Thermal Death Tube (TDT)
method. The effect of previous heat shock (110 ºC/15’) on resistance to HPH was also
evaluated by heating of the inoculated milk prior to homogenization at 300 MPa. To measure
the effect of inlet temperature in the spore inactivation, skin milk inoculated was pre heated to
45ºC and then subjected to 300MPa. Additionally, the effect of successive treatment of
homogenization at 300MPa was evaluated by milk recirculation on the homogenizer until it
reached the equivalent to 16 times. In all tests performed the count of spores suspension were
determined before and after the treatment, in order to evaluate the spores reduction (NDR),
through equation 1. Data were statistically evaluated through variance analysis (ANOVA) and
average test (Tuckey) using the software STATISTICA 5.0.

NDR log initial _ spores  log spores _ after _ treatment   Equation 1

RESULTS & DISCUSSION

The results showed no inactivation of spores at 100, 200 and 300MPa, also the NDR
determined for B. stearothermophilus were negative, indicating a possible activation of the
spores, as previously described using high hydrostatic pressure (HHP) [2]. Considering that
HPH was not enough to inactivate the spores in milk, it was tested if the HPH could sensitize
the spores heat, due to its activation. This effect was previously observed by using HHP [3],
however no changes in D-value and z-value were observed for microoganisms after
homogenization at 300MPa. Therefore, the HPH was not able to promote sublethal injuries or
activation in theses spores in milk, which is in accordance with others authors [4,5].

11th International Congress on Engineering and Food - Athens, Greece, 2011 1661
The previous thermal germination of spores were tested but, again no significant reductions of
B. stearothermophilus and C. sporogenes were observed, indicating that HPH up to 300MPa
was also not able to inactivate germinated spores. The use of HPH at high temperatures (84ºC)
also was not effective against the tested spores. The last alternative to makes viable the use of
HPH to inactivate B. stearothermophilus (figure 1) and C. sporogenes spores was the multiple
treatments at 300MPa (multiple passes). Again, low efficacy was observed in the inactivation
of the spores.

1,00E+06 0,8
0,6
Spores counts / ML

0,4
0,2

NDR
1,00E+05
0
-0,2
-0,4
1,00E+04 -0,6
0 5 10 15 20
Number of Passes
Spores counts/ mL NDR

Figure 1. Effect of multiple passes at 300MPa on the inactivation of B. stearothermophilus spores

CONCLUSION
It was concluded that HPH process was not able to reduce or the tested spores in skim milk at
the pressure up to 300MPa, even when applied multiple treatment at high pressure of 300MPa,
or associated with mild heat treatment. Therefore, this process is not suitable for commercial
purposes aiming to inactivate spores in milk.

ACKNOWLEDGEMENTS
The authors thank FAPESP for financing project 2001/06872-2, 2004/07074-0, 2005/53668-2.

REFERENCES
[1] Feijoo, S.C.; Hayes, W.W.; Watson, C.E.; Martin, J.H. (1997). Effects of Microfluidizer Technology
on Bacillus licheniformis spore in ice cream mix. Journal of Dairy Science, 80: 2184 –2184
[2] Gould, G. W.(1973). Inactivation of Spores in Food by Combined Heat and Hydrostatic Pressure.
Acta Alimentaria. 2 (4), 377-383.
[3] Mills, G.; Earnshaw, R.; Patterson, M. F. (1998). Effects of high hydrostatic pressure on Clostridium
sporogenes spores. Letters in Applied Microbiology,26, 3, 227-230.
[4] Wuytack, E.Y; Diels, A.M.J; Michiels, C.W. (2002). Bacterial inactivation by high-pressure
homogenization and high hydrostatic pressure. Int. J. Food Microb., 77: 205-212.
[5] Briñez, W.J.; Roig-Sagués, A.X.; Herrero, M.M.H.; López, B.G (2007). Inactivation of
Staphylococcus ssp. strains in whole milk and orange juice using ultra high pressure homogenization
at inlet temperatures of 6 and 20ºC. Food Control, 18: 1282-1288.

1662
 Effect of ultra high pressure homogenization on alkaline phosphatase and

lactoperoxidase activity in raw skim milk

Cláudia R. G. PINHO; Mark A. FRANCHI; Alline A. L. TRIBST; Marcelo CRISTIANINIa

a
Department of Food Technology (DTA), School of Food Engineering (FEA), University of Campinas
(UNICAMP), Campinas, SP, Brazil (olecram@fea.unicamp.br)

INTRODUCTION
High pressure homogenization (HPH) is an alternative food processing technique. It preserves
sensory and nutritional food characteristics due to low heating, being interesting to guarantee
the safety of thermo-labile food [1]. The inactivation of native biota and also of some
microorganisms intentionally added to milk [1, 2] suggested that HPH in milk promotes effect
similar to thermal pasteurization, which is usually conduced at 72.5oC/15seconds. Alkaline
phosphatase (AP) and lactoperoxidase (LP) are enzymes used as thermal pasteurization
indicator, since AP is thermo-labile and inactivated by pasteurization while LP is heat resistant.
The present work aimed to evaluate the residual activity of AP and LP after treatment by HPH,
to determine if these enzymes can be also used as HPH indicator.

MATERIALS & METHODS

The assays were performed by treating raw milk in a high pressure homogenizer Stansted,
model FPG 7400H:350 at pressures of 100, 150, 200, 250, 270 and 300 MPa. The tests were
performed in triplicate. The activity of AP was determined colorimetrically based on the
phenol liberation and the activity of LP was determined spectrophotometrically based on the
oxygen liberation and reaction with guaiacol. Enzymatic activities were measured before and
after each pressure treatment and the tests were performed in duplicate.

RESULTS & DISCUSSION

The enzymes inactivation can be associated to HPH effects or to the heating that occurred on
milk due to the high shear that occurs during the decompression. Considering the thermal
effect, it was observed that at 270MPa and 300MPa the reached temperature were 74ºC and
80,2ºC, respectively. Considering the D and z-values of both enzyme, in HPH above 270MPa it
probably occurred thermal inactivation of AP and, at 300MPa, partial inactivation of LP.
Figure 1 shows the residual activity of AP and LP. The inactivation of AP is probably due to
heating effect associated to HPH, as previously observed by other authors [3]. Considering that
the majority of HPH process to milk pasteurization suggests application of pressure above
250MPa [2], the activity of AP can be used as an indicative of inadequate HPH pasteurization.
The results of LP indicated that only treatment at 300MPa was able to partially reduce the
enzymatic activity, making possible the use of this enzyme as an indicator of excessive
treatment. It is not possible to determine if the partial inactivation observed at 300MPa is
consequence of the homogenization or the heating associated to the process, however, previous
work had observed that the inactivation of LP by HPH is higher than the obtained with the
compatible heat treatment, suggesting an additive effect of the HPH and the generated heat [3].
Results obtained for LP at pressures of 100 and 250MPa showed incease of the enzymatic

11th International Congress on Engineering and Food - Athens, Greece, 2011 1663
activity. Recent results have been indicating that relative low pressures can be activate
enzymes due to changes on the spatial configuration, with the enhancement of active sites
exposure [4].

160
140
Enzymatic Activity (%)

120
100
80
60
40
20
0
0 50 100 150 200 250 300
Pressure (MPa)

Alkaline Phosphatese Lactoperoxidase

Figure 1. Residual activity of Lactoperoxidase and AP after high pressure homogenization

CONCLUSION
It was concluded that AP presented high sensitivity to homogenization process than LP,
however, this sensitivity probably is associated to the heat generated during the process, since
the AP is thermo-labile. Also, for the LP it was concluded that just 300MPa were able to
promote some reduction on the enzymatic activity, which can be attributed to the sum of the
effects of pressure and the temperature reached during the treatment. Considering the results it
was concluded that AP and LP activities can be used as an indicative of pressure level reached
during HPH.

ACKNOWLEDGEMENTS
The authors thank FAPESP for financing project 2001/06872-2, 2004/07074-0, 2005/53668-2.

REFERENCES
[1] Lanciotti, R.; Patrignani, F.; Iucci, L.; Saracino, P.; Guerzoni, M.E. (2007). Potential of high pressure
homogenization in the control and enhancement of proteolytic and fermentative activities of some
Lactobacillus species. Food Chemistry, 102: 542-550. [2] Thiebaud, M.; Dumay, E.; Picart, L.; Guiraud,
J. P.; Cheftal, J. C. (2003). High-pressure homogenization of raw bovine milk. Effects on fat globule size
distribution and microbial inactivation. International Dairy Journal, 13(6): 427-439. [3] Datta, N.; Hayes,
M. G.; Deeth, H. C.; Kelly, A. L (2005). Significance of frictional heating for effects of high pressure
homogenization on milk. Journal of Dairy Research, 72 (4): 393-399. [4] Liu, W.; Liu, J.; Liu, C.;
Zhong, Y.; Liu, W.; Wan, J. Key, S. (2009). Activation and conformational changes of mushroom
polyphenoloxidase by high pressure microfluidization treatment. Innovative Food Science and Emerging
Technologies, v. 10, p. 142–147.

1664
 Changes in texture, structure and pectin of peach during pressurization, heating or

processing of high-pressure-induced and heat-induced jam

Hiroko Kuwadaa, Yuri Jibub, Keiko Nakamurab, Mayumi Tabuchib, Ai Teramotoc, Kayoko Ishiia, Yasumi
Kimuraa, Michiko Fuchigamia

a
Department of Nutrition and Life Science, Fukuyama University, Fukuyama, Japan
(kuwada@fubac.fukuyama-u.ac.jp; ishii@fubac.fukuyama-u.ac.jp; kimura@fubac.fukuyama-u.ac.jp;
fuchigam@fubac.fukuyama-u.ac.jp)
b
Department of Nutritional Science, Okayama Prefectural University, Soja, Japan (yjibu@fhw.oka-
pu.ac.jp; keikotin@yahoo.co.jp; tabuchi@fhw.oka-pu.ac.jp)
c
Department of Health and Nutrition, Kanto Gakuin University, Yokohama, Japan (teramoto@kanto-
gakuin.ac.jp)

INTRODUCTION
Jam is prepared by boiling fruit with sugar. During boiling, two processes, pectin extraction
and jam manufacture, are performed. Heat-induced-jam has some faults such as off-flavor and
deterioration of food components, nutrients and especially color. However, high pressure can
produce jam without heating because pressurization accelerates hydrogen bonds between
pectin macromolecules. It does not greatly change food color during processing [1]. In
previous research high-methoxyl pectin was extracted by soaking in 0.01N HCl solution (pH
2.0) at 35ÛC due to remove of Ca2+, the vegetables softened [2] [3]. This extraction method of
pectin was used for softening the peel by using citric acid instead of HCl for yuzu marmalade
(Kuwada et al., 2010). Consequently, the peel softened. Although peach is softer than yuzu
peel, soaking in citric acid solution may be useful for the extraction of pectin from peach. Thus,
the objectives of this study are to research the relationship between pectin and the softening of
peach by soaking in citric acid solution, pressurizing or heating, and to establish a process for
HP-jam and compare it with H-jam.

MATERIALS & METHODS

Sample preparation
Peach (Prunus persica L.) was diced into 1 cm pieces. The vacuum-packed pieces were
pressurized for 30 min at 500 MPa at room temperature using a Dr. Chef high pressure food
processor (Kobe Steel Ltd.) or boiled in hot water for 10 min. Also, pieces were soaked in
citric acid solutions (pH 2.0, 2.2 or 2.5) for 24 hrs at 35ÛC.

Texture and structure measurements and extraction of pectin

Texture and histological structures were measured by a Rheoner (RE-33005) and a cryo-
scanning electron microscope (S-4500). Pectic substances were extracted successively into five
reagents; distilled water, 0.01N HCl, 0.1M sodium acetate buffer, 2% sodium
hexametaphosphate solution, and 0.05N HCl. These extracts were designated as WSP, PA, PB,
PC and PD, respectively.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1665
Jam preparation and methods for the quality evaluation of jams
Eight kinds of peach jam (65% sugar, pH 2.0 or pH 2.2, and 50% or 60% sugar, pH 2.5) were
produced. Peach dices were soaked in citric acid then sucrose was added. They were vacuum-
packed, then pressurized for 30 min at 500 MPa (HP-jam) or boiled for 10 min (H-jam),
respectively. The Color and rheology of jams were measured using a spectrophotometer (ZE-
6000) and a Rheosol-G3000, respectively. Sensory evaluation of peach jam was performed
using a five point scale. The color, transparency, flavor (smell), texture, sweetness, sourness,
mouthfeel, total taste and preference of jam were compared.

RESULTS & DISCUSSION

Changes in texture, structure and pectin composition of peach during soaking, pressurizing or
heating
Firmness of the peach decreased greatly when soaked at pH 2.0 > heated > soaked at pH 2.2 or
2.5 > pressurized, respectively. Middle lamella of cell walls separated more from heating for
10 min than soaking at pH 2.5. However, they did not separate when pressurized. About 88%
of the peach pectin was water-soluble-pectin (WSP) of low molecular weight and high-
methoxyl-pectin (PA), while low-methoxyl pectin (PB, PC and PD) was slight. The pectin did
not change during pressurization. However, pectin degraded during heating; consequently the
middle lamella separated. The pH value of raw peach was 4.46. Since pectin does not degrade
through ȕ-elimination by pressurization or by heating at pH 4, it might degrade through
hydrolysis by enzyme.

The quality of high-pressure-induced and heat-induced peach jams

The steady-flow viscosity of H-jam was slightly higher than HP-jam. As the pH value of citric
acid solution was higher and sugar content was lower, viscosity decreased. As pH values
became lower, L-, a-, b-values of the jam became higher, and thus, jam became pinker. L-, a-,
b-values of HP-jam were higher than H-jam. This suggests that the amount of anthozyan
(pigment of peach) was maintained by pressurizing but decreased by heating. Color and flavor
of HP-jam were better than the H-jam. However, there was no significant difference in sensory
evaluation between HP- and H-jams.

CONCLUSION
Raw peach contained about 0.3~0.4% pectin, therefore an addition of 0.6% pectin was needed
for pressure-induced (HP) jam.

REFERENCES
[1] Hayashi, R. 1989. Use of high pressure in food (pp. 1-30), San’ei Press, Kyoto.
[2] Fuchigami, M. & Okamoto, K. 1984. Fractionation of pectic substances in several vegetables by
successive extraction with dilute hydrochloric acid and acetic buffer solutions. Journal of Japanese
Society for Nutrition and Food Science, 37(1), 57-64.
[3] Fuchigami, M. 1987. Relationship between pectic compositions and the softening of the texture of
Japanese radish roots during cooking. Journal of Food Science, 52(5), 1317-1320.
[4] Kuwada, H., Jibu, Y., Teramoto, A., &, Fuchigami, M. 2010. The quality of high-pressure-induced
and heat-induced yuzu marmalade. High Pressure Research, 30(4), 547-554.

1666
Effects of high pressure with the addition of sugar-alcohol on the improvement in texture

and structure of frozen egg custard gel

Ai. Teramotoa, Yuri Jibub, Hiroko Kuwadac, Yasumi Kimurac, Kayoko Ishiic, Michiko Fuchigamic

a
Department of Health and Nutrition, Kanto Gakuin University, Yokohama, Japan (teramoto@kanto-
gakuin.ac.jp)
b
Department of Nutritional Science, Okayama Prefectural University, Soja, Japan (yjibu@fhw.oka-
pu.ac.jp)
c
Department of Nutrition and Life Science, Fukuyama University, Fukuyama, Japan
(kuwada@fubac.fukuyama-u.ac.jp, kimura@fubac.fukuyama-u.ac.jp, ishii@fubac.fukuyama-u.ac.jp,
fuchigam@fubac.fukuyama-u.ac.jp)

INTRODUCTION
Egg custard gel is formed by heat-induced aggregation of egg protein molecules. A smooth
taste is considered to be highly important in egg custard gel. However, the texture of egg
custard gel when frozen at atmospheric pressure (0.1 MPa) is unsuitable for consumption
because it is spongy. A non-freezing region (liquid phase) below 0ÛC exists under high
pressure [1]. When water is pressurized at 200 MPa at -20ÛC, it does not freeze. However,
when the pressure is released, water freezes quickly by pressure-shift-freezing [2]. If this is
done to a frozen gel with high water content, damage to the gel may be reduced. The objective
is to determine the effects of pressure-shift freezing with the addition of sorbitol or maltitol on
the improvement in texture and structure of frozen egg custard gel.

MATERIALS & METHODS

Three packs of egg custard gel with 0% (sugar-free) and 5% sorbitol or maltitol were put into a
pressure vessel and immediately pressurized for 90 min at -20ÛC at 0.1 ~ 686 MPa using a high
pressure food processor (Kobe Steel Ltd.) [3] [4] [5]. After decompression, gel was stored for 1
day at -30ÛC, then thawed at 20ÛC. Changes in the temperature of samples during freezing were
compared. Also, the texture and structure of high-pressure-frozen gel were compared with
untreated and frozen (in freezers at -20ÛC, -30ÛC or -80ÛC) gels using a creepmeter (Rheoner,
RE-33005, Yamaden Ltd.) and a cryo-scanning electron microscope (S-4500, Hitachi Ltd.),
respectively.

RESULTS & DISCUSSION

During pressurization at 100, 500, 600 and 686 MPa, the phase transition of liquid to ice
occurred in sugar-free-gel. However, the sugar-free and 5% sugar-samples did not freeze at -
20ÛC at 200 ~ 400 MPa and 200 ~ 500 MPa, respectively, but phase transition occurred when
the pressure was released. It was found that the amount of syneresis from gel pressure-shift-
frozen at 200 ~ 400 MPa was smaller than that frozen at other pressures.
When gel was frozen at 100, 500, 600, or 686 MPa, rupture stress and strain increased.
However, that of pressure-shift-frozen gels did not change greatly. Furthermore, change in the
rupture stress of gels was reduced with the addition of 5% sugar.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1667
The size of the ice crystals in gel pressure-shift-frozen at 200 ~ 400 MPa was smaller than that
in the gel frozen at other pressures. A large number of small round ice crystals formed
homogenously throughout the pressure-shift-frozen gel.

CONCLUSION
The gels with 0% and 5% sugar-alcohol (sorbitol or maltitol) did not freeze at about -20ÛC
during pressurization at 200 ~ 400 and 200 ~ 500 MPa, respectively. Thus, depression of the
freezing point in 5% sugar-alcohol-gel was greater than the 0% sugar-alcohol-gel. When
pressure was released, the supercooled gel froze quickly by pressure-shift-freezing and small
ice crystals of a granular shape were dispersed. Therefore, stress and strain of pressure-shift-
frozen gel did not change greatly, while, the addition of sorbitol or maltitol decreased the size
of ice crystals. Thus, pressure-shift-freezing and the addition of sugar-alcohol appeared to be
effective in improving the quality of frozen egg custard gel.

REFERENCES
[1] Fretcher, H. N. 1970. The Chemical Physics of Ice. (p. 271), Cambridge Univ. Press, Bristol.
[2] Kanda, Y., Aoki, M. & Kosugi, T. 1992. Freezing of Tofu (Soybean Curd) by Pressure-shift
Freezing and its Structure, Journal of the Japanese Society for Food Science and Technology, 39(7),
608-614.
[3] Teramoto, A., Ogawa, N. & Fuchigami, M. 2001. Structural and Textural Quality of High-pressure-
frozen Egg Custard Gel as Affected by Sucrose. In, Proceedings of the Eighth International
Congress on Engineering and Food, Lancaster.
[4] Teramoto, A. & Fuchigami, M. 2002. Effects of High Pressure and Salts on Frozen Egg Custard Gel.
In Trends in High Pressure Bioscience and Biotechnology (Hayashi, R. ed., pp. 469-476), Elsevier
Science B.V., Amsterdam.
[5] Teramoto, A., Jibu, Y. & Fuchigami, M. 2006. Structural and Textural Quality of Pressure-shift-
frozen Egg custard Gel as Affected by Glucose, Trehalose or Sucrose. Journal of Cookery Science
of Japan, 39(3), 194-202.

10 10
Sugar-free
Ruptue stre ss (x10 N/m )

5% Sorbitol
2

8 8
5% Maltitol
4

6 6

4 4

2 2

0 0 -20
Control0.1 100 200 340 400 500 600 686 -30 -80

Pressurized (MPa) at about -20䉝 Frozen in freezers (䉝 )

Figure 1. Effects of high pressure and sugar-alcohol on rupture stress of frozen-thawed egg custard gel.

1668
 Process variables study on supercritical CO2 extraction of Brazilian cherry seeds

(Eugenia uniflora L.) rich in bioactive volatile

Débora Nascimento e Santosa, Larissa Lima de Souzaa, Nilson José Ferreiraa, Alessandra Lopes de
Oliveiraa*

a
College of Animal Science and Food Engineering ou Faculdade de Zootecnia e Engenharia de
Alimentos, University of São Paulo, Pirassununga, Brazil (alelopes@usp.br)

INTRODUCTION
The Brazilian cherry or Pitanga (Eugenia uniflora L.) is a widely distributed plant in Latin
America, and it presents numerous benefits to human health. The Pitanga leaves and fruits are
used in home medicine in several country regions because they are considered stimulant,
febrifuge, aromatic, antirheumatic, antidiarrheal and, the alcoholic extract is used in bronchitis,
coughs, fevers, anxiety, hypertension, and worm diseases [1]. The Pitanga seed is currently a
waste from fruit process and has no proper commercial but was found a high antioxidant
potential, twice as large as various pulps [2]. The supercritical fluid extraction (SFE) has been
recognized as a promising process for application in food, pharmaceutical and cosmetic
industries as having high selectivity, low extraction times and for not using toxic organic
solvents [3]. Our objectives were to obtain seeds extracts from Brazilian cherries using
supercritical CO2, to quantify the total phenolic compounds in these extracts and perform the
composition analysis by GC/MS.

MATERIALS & METHODS

Brazilian cherries collected were selected, washed, dried and ground in mill hammer type. To
characterize the sample, an analysis for moisture content, fat and particle size was performed.
The extraction procedure is based on promoting contact among the ground seeds with
supercritical CO2 under set conditions of pressure (P) and temperature (T), defined according
to CCD 22 with P and T as independent variables (Table 1). In extracts obtained were
quantified the total phenolic compounds and the volatile composition of the extracts was
analyzed by gas chromatography coupled with mass spectrometry (GC/MS).

RESULTS & DISCUSSION

The samples of crushed dry seeds showed moisture content of 12.73 ± 0.2%. The average value
of ethereal extract was 0.52 ± 0.09% and the means of particle size was 0.48 mm. Table 1
shows the results of yield and concentration of phenolics compounds for each essay based on
the experimental design.
Highest yields are obtained with higher P and therefore it is possible to use central values of T
in the process that could ensure the integrity of thermosensitive substances. With respect to
phenolic compounds (TPC) for the variables studied in their respective intervals, no significant
effect was observed. The range which has achieved the highest amounts of phenolic
compounds is in middle ranges of T and P. The chromatographic analysis indicated that major

11th International Congress on Engineering and Food - Athens, Greece, 2011 1669
volatile constituents are the sesquiterpenes, germacrone and germacrene B, which are also
present in the Pitanga fruit and leaves essential oils [4].

Table 1. Matrix of CCD 22 (including three central points) to study the effect of P and T on the yield and
TPC.
Coded Variables Variables TPC
Yield TPC
Essay 2 o 2 o (mg GAE×100-1 g
P (Kgf/cm ) T ( C) (P) (Kgf/cm ) T ( C) (%) (ppm)
seeds)
1 -1 -1 110 35 0.28 49.92 0.014
2 +1 -1 240 35 0.46 42.76 0.019
3 -1 +1 110 55 0.23 61.89 0.014
4 +1 +1 240 55 0.47 17.95 0.008
5 -1.41 0 83 45 0.16 39.88 0.006
6 +1.41 0 267 45 0.42 48.93 0.021
7 0 -1.41 175 31 0.39 23.28 0.009
8 0 +1.41 175 59 0.46 67.09 0.031
9e 0 0 175 45 0.44 73.89 0.033
10e 0 0 175 45 0.40 44.54 0.018
11e 0 0 175 45 0.48 61.72 0.030
e = central points

CONCLUSION
The study of operational conditions for SFE of Brazilian cherry seeds showed that the best
yields can be obtained at high pressures and low temperatures. For the phenolic compounds it
was noted that P and T did not influence this content when the CCD was applied. The major
volatile compounds found were the sesquiterpenes germacrene B and germacrone, with
biological action and potential application in pharmaceutical and cosmetic industry.

REFERENCES
[1] Consolini A.E. & Sarubbio M.G. 2002. Pharmacological Effects of Eugenia uniflora (Myrtaceae)
Aqueous Crude Extract on Rat’s Heart. Journal of Ethnopharmacology , 81, 57-63.
[2] Bagetti, M., Facco, E.M.P., Rodrigues, D.B., Vizzotto, M. & Emanuelli, T. 2009. Antioxidant
Capacity and Composition of Pitanga Seeds, Ciência Rural, 39(8), 2504-2510.
[3] Serra, A.T., Seabra, I.J., Braga, M.E.M., Bronze, M.R., Sousa, H.C. & Duarte, C.M.M. 2010.
Processing Cherries (Prunus avium) Using Supercritical Fluid Technology. Part 1: Recovery of
Extract Fractions Rich in Bioactive Compounds. The Journal of Supercritical Fluids, 55, 184-191.
[4] Costa, D.P., Alves Filho E.G., Silva, L.M.A., Santos, S.C., Passos, X.S., Silva, M.R.R., Seraphin, J.C.
& Ferri, P.H. 2010. Influence of Fruit Biotypes on the Chemical Composition and Antifungal Activity
of the Essential Oils of Eugenia uniflora Leaves. J. Braz. Chem. Soc., 21(5), 851-858.

1670
 High hydrostatic pressure (HHP) microbial kinetics in orange comminuted

Vinicio Serment-Moreno a, Zamantha Escobedo-Avellanedab, Jorge Welti-Chanesc

a
Instituto Tecnológico y de Estudios Superiores de Monterrey, Monterrey, México(vsermentm@gmail.com)
b
Instituto Tecnológico y de Estudios Superiores de Monterrey, Monterrey, México(zamantha17@yahoo.com.mx)
c
Instituto Tecnológico y de Estudios Superiores de Monterrey, Monterrey, México(jwelti@itesm.mx)

INTRODUCTION
An orange derived paste can be obtained by mixing several of its components, where juice is the major
contributor. This product, known as comminuted, can be used as a refreshing drink base to enhance nutritional
and sensorial characteristics. However, as orange peel is used for comminuted elaboration, microbial
contamination may also be favoured. Orange fruit spoilage is frequently related to acidic fermentative
microorganisms which can be readily inhibited by freezing or applying heat treatment [1]. Nevertheless,
orange peel is a great source of nutraceutic compounds [2] which can easily be degraded by extreme
temperature exposure. The present study proposes a high hydrostatic pressure treatment (HHPT) to achieve
microbial safety in orange comminuted.

MATERIALS & METHODS

Orange comminuted was prepared by grinding orange components in the following proportions: juice 71.6%
(w/w), pericarp 12.4% (w/w) and outer layer 16.0% (w/w). Fifty grams of comminuted were vacuum sealed in
polyethylene bags and processed with a Welch 2L Food Processor (Avure Technologies). Samples were
pressurized from 100-414 MPa during 1-4 min at room temperature. Aerobic counts were performed by
poured plate technique for both nontreated and processed samples. Treatment efficiency was determined by
the microbial logarithmic reduction.

RESULTS & DISCUSSION

Comminuted orange presented a moderate initial microbial load (103-105 CFU/g). Low pressures (100-200
MPa) barely accounted to reduce 1.5 log cycles. Bacterial populations were stabilized as pressure and
treatment duration levels increased and a “tail effect” trend can be observed. Meawhile, treating comminuted
samples with 300 and 414 HHP resulted effctive to eliminate microbial load (<10 CFU/g) by treating samples
with at least 4 and 2 min respectively. However, [3] achieved complete E. coli inhibition by just applying
CUT (Come Up Time, which accounts for the amount of time the equipment needs to reach the desired
operating pressure) at 276 MPa and higher pressures.
It is also important to denote the curvature concavity inversion when applying 300 MPa. This adjustment may
indicate population variation, or that their adaptability towards the stress conditions of high pressure
processing have gone beyond its limit and can no longer be accumulated. As a result, treatment efficiency was
remarkably increased as both pressure and time levels augmented.
Calculated inactivation constants (k [=] min-1) for a first order kinetics ranged from 0.13 through 1.41
(R2=0.78-0.91) are shown in Table 1. Guerrero-Beltrán and others [4] reported k values of 0.51 and 1.53 to
inactivate aerobic bacteria in orange juice at 300 and 400 MPa respectively. Decimal reduction times obtained

11th International Congress on Engineering and Food - Athens, Greece, 2011 1671
at 300 and 400 MPa (D = 1.99 and 0.69 min) were in accordance with those of [5] for native microflora in
Hamlin orange juice treated at 350, 400 and 450 MPa (D = 1.32 min, 0.47 min and 0.18 min respectively).
Bacterial population shows moderate resistance toward pressure exposure as noted by z = 317 MPa. However,
z value calculated diverted largely when compared with 85-103 MPa obtained by [3, 5] or 556 MPa [4]. Result
variation may be due to orange variety, processing conditions, peel addition or come up time consideration for
kinetic calculations, since D values are similar between them.
Nonetheless, the first order kinetics was only followed at 100 MPa, since all greater pressures exhibit a
resistant remaining population. Weibull equation (1) was also applied to describe the experimental data
generated. This model adequately described kinetics from 200-414 MPa, especially in the nonlinear data
range. All curves presented downward concavities as noted by n<1 (Table 1) with the exception of 300 MPa.
This could indicate that damage is being accumulated until a pressure threshold is reached (n=1, between 200-
300 MPa). Extending treatments beyond this critical value can result in diminished bacterial resistance and the
already noticed curve concavity inversion. Both parameters seem to be pressure dependant with no obvious
tendency, manifesting peak values for the upward concavity curve (300 MPa).
െ Ž‘‰ ܵ ൌ െሺ‫ݐ‬Ȁߜሻ௡ (1)

Table 1. First order (left) and Weibull (right) inactivation kinetic parameters for pressure treated orange comminuted
P(MPa) k(min-1) D(min) R2 į(min) n R2
100 0.13 7.42 0.89 1.59 -0.84 0.25
200 0.33 3.02 0.78 1.00 0.31 1.00
300 0.50 1.99 0.85 8.74 1.28 0.74
414 1.41 0.69 0.91 0.74 0.77 0.80

CONCLUSION
High hydrostatic pressure processing is definitely suitable for orange comminuted pasteurization. Orange peel
addition for comminuted preparation may imply a greater microbial initial population, but does not represent
an important safety threat. Inactivation kinetics for comminuted do follow a first order inactivation model, and
seems very closely related to those describing microorganism inhibition for different orange juice varieties.

REFERENCES
[1] Zook C.D., Parish M.E., Braddock R. J. & Malaban M. O. 1999. High pressure inactivation kinetics of Saccharomyces
cerevisiae ascospores in orange and apple juices. Journal of Food Science 64(3), 533-535.
[2] Tripoli E., La Guardia M., Gimmanco S., Di Majo D. & Giammanco M. 2007. Citrus flavonoids: Molecular structure,
biological activity and nutritional properties: A review. Journal of Food Chemsitry 104(4), 466-479.
[3] Guerrero-Beltrán J.A., Barbosa-Canovas G. V. & Welti-Chanes J. 2011. High hydrostatic pressure effect on natural
microflora, Saccharomyces cerevisiae, Escherichia coli, and Listeria Innocua in Navel orange juice. International
Jouranl of Food Engineering. Manuscript 2166.
[4] Dogan C. & Erkmen O. 2004. High pressure inactivation kinetics of Listeria monocytogenes inactivation in broth, milk
and peach and orange juices. Journal of Food Engineering 62, 47-52.
[5] Parish M. E. 1998. High pressure inactivation of Saccharomyces cerevisiae endogenous microflora and
pectinmethylesterase in orange juice. Journal of Food Safety 18, 57-65

1672
 Research Development of Ultra-High Pressure Processing on Fruit Juice
Wu Han, Zhang Yunchuan, Han Qinghua, Zhao Youbin
Chinese Academy of Agriculture Mechanization Sciences, Beijing, china (paley366@sina.com)

INTRODUCTION
Ultra high pressure (UHP) technique subjects liquid and solid food to pressures between 100
and 1000MPa with exposure times ranging from a few seconds to over 30min, which is on the
aim of microbial inactivation. As a non-thermal preservation technique UHP technology can
ensure high quality food products. The use of UHP for food processing has been developed
extensively and has a broad application prospect. Since beginning, the research about the UHP
technique was mainly used for food preservation. And the sterilization is the primary intention
of research about the UHP technique all the time, where there are three primary factors of
preservation treatment: sterilization, enzyme inactivity and the quality. In addition the
development of UHP technique research is based on the breakthrough of the UHP equipment
technique.
STERILIZATION OF UHPP ON FRUIT JUICE
The principle of sterilization about UHP technique is inactivation of microorganism by
pressure. Recently many researches about sterilization of the UHP were carried out. Their
materials were as follow: orange juice, apple juice, tomato juice, watermelon juice, pear juice,
Hami melon (Cucumis Melo L.) juice and so on. The researches indicated that the sterilization
effects of fruit juice by the UHPP concerned with the pressure size, the time pressed, the
quantity of microorganism in fruit juice, the microorganism type, the material PH, the
temperature of processing and so on many factors. At present the prominent problem of the
UHP technology is the resistance of pressure- resistant spore in sterilization. It is some
bacterial spores that still could survive under the pressure of 1200MPa[1]. Therefore
experiments were carried out on various materials by the cooperated processing methods of
sterilization, which are the UHPP and the temperature, the PH value or the ultrasonic wave
processing work together. And good effects have been obtained[2]. It is difficult to carry out the
cooperated processing methods outside, where the UHPP needs a strict pressurizing in
processing tank. So finding cooperated processing methods is still the direction of research in
recent years.
ENZYME INACTIVITY OF UHPP ON FRUIT JUICE
The UHP treatment mainly takes effects on the tertiary structures of enzyme, which make the
inhibition of enzyme activities. Researches indicate that the inactivation of enzyme is the
causation of secondary bonus’s breakage and protein structure destroyed [3]. It is similar with
the UHP sterilization which has many influencing factors, the enzyme inactivity effects of fruit
juice by the UHPP concerned with the pressure size, the time pressed, the quantity of enzyme
in fruit juice, the enzyme type, the material PH, the temperature of processing, the enzyme
inhibitory and so on many factors. The enzyme inhibitory by the UHPP has its particularity. It
will activate some enzyme, when many enzyme activities were inhibited [4]. Some enzymes,
such as pectin methyl esterase, peroxidase, polyphenol oxidase, or lipoxygenase, are specially
resistant to pressure. The peroxidase has the strongest resistance to pressure, which might
maintain 90% activeness under the UHP treatment of 30min at 600MPa and 60 oC [4]. In view

11th International Congress on Engineering and Food - Athens, Greece, 2011 1673
of the fact that some enzymes cannot inactivated completely with simple high pressure, study of new
technical coordination UHPP is still one of solution methods.
QUALITY OF UHPP ON FRUIT JUICE
In recent years, there are a lot of researches about processing quality on fruit juice by the UHP treatment,
which concern the colour, texture, flavor, nutrient of fruit juice. Many researches indicate that the fruit
juice treated by UHPP may have storage more than a half year, moreover it has much less change in
nutrient, flavor and colour compared with the heat-treatment method. This is the latent superiority of the
UHPP in fruit juice processing. a part of basement researches about the UHPP are studies about the
changes of the colour, texture, flavor and nutrient on single material juice by UHPP compared with the
heat-treatment method.
EQUIPMENT OF UHPP

The UHP technique has been used in food processing until Japan mad the first high pressure food
processing testing facility at the end of 1980s. At present, America and Japan, and the others have got
achievement of research and development, standardization and mass production in high pressure
processing facility, such as the Flowing Company and Wenger Company of America, Meidi-ya food
Company of Japan, which all has its own characteristic productions.The processing capacity of high
pressure processing of food facility has got 275mpa, 24.6kg/min serialization production. In china, the
leading high pressure processing of food facility manufacture-----Baotou High Pressure Technique Co;
LTD that the facility capacity in sold is only 15L at the maximum working pressure 800mpa; and the
facility capacity is just 500L at the maximum working pressure 600mpa. Currently, in china it is difficult
in application and extension of the UHPP products of high pressure technique. But the industrial
development of UHPP technique has unlimited potential after reducing the cost of equipment and
processing.
Final Remarks: Ultra-high pressure processing (UHPP) technique is a new technique in food cold
sterilization, which has a broad application prospect. The ultra-high-pressure processing technique has
already used and applied in production on a large scale, as the good processing quality makes
destructiveness very small in the nature nutrient component, flavor, and the color which turns brown.
However, the effect of UHPP in sterilization of bacterial spores and inactivation of peroxidase is not
good, and which cannot be improved simply by enhances the pressure. As the research of next step, we
can carry on the work as follows. Some enhancement methods of Ultra-high-pressure processing, such as
magnetization and ultrasonic wave, are carried on fruit juice. Research of dynamics model about
Sterilization and enzyme inactivity of Ultra high pressure processing on fruit juice are also needed. In
shot, UHPP is a promising technology that could eventually replace many heat treatments in food
processing.

REFERENCES

[1] Zeng QinmeiˈPan JianˈXie Huiminˈet alˊStudy on ultra-high pressure treatment of microflora sterilization of
strawberry juice [J]ˊFood Scienceˈ2004ˈ25 (1)˖31~34. [2] Zhang YongˈDuan XuchangˈLi Shaofengˈet alˊInfluence
factors of sterilization and deactivation enzyme of ultra-high pressure processing [J] ˊ Food Research and
Developmentˈ2007ˈ28 (7)˖140~143ˊ[3] Rastogi.N.K, Eshtiaghi.M.N, Knorr.DˊEffects of peroxide and high pressure
treatment on the reduction of peroxidase and polyphenoloxidase activity un red grapes[J] ˊ Food
Biotechnology,1999,13(2)˖195~208ˊ[4] Murao, S, et alˊEnhancement of acitivities of cellulases under high
hydrostatic pressure[J]ˊBiosci. Biotech. Biochem.,1992,56(8)˖1366~1367ˊ

1674
 Effects of high hydrostatic pressure on antioxidant activity, mineral and starch content

and bioaccessibility, in apple (Granny smith)

Vilbett Briones-Labarcaa, Gabriela Venegas-Cubillosa, Susana Ortiz-Portillaa, Marcelo Chacana-Ojedaa,

Hector Maureirab

a
Department of Food Engineering, Universidad de La Serena, La Serena, CHILE, vbriones@userena.cl
b
Central Laboratory Analysis, Universidad de La Serena, La Serena, CHILE.

INTRODUCTION
The diet is important for human health because it is associated with the morbidity and mortality
in the chronic diseases such as cardiovascular disease, cancer, hypertension and obesity. The
effects of the food matrix on the bioavailability or bioaccessibility of antioxidant minerals and
starch have not been examined in much detail. Direct interactions between this component and
some components of food, such as binding to proteins and polysaccharides, can occur, and
these interactions may affect digestion and absorption. The objective of this study was to study
the effect of high pressure on the bioaccessibility of specific nutrients (antioxidant, minerals
and starch) in apple and to establish processing conditions that maximize the health benefits.

MATERIALS & METHODS

Apple samples were pressurized at 500 MPa during 2, 4, 8 and 10 min. Free radical scavenging
activity of the samples were determined using the 2,2,-diphenyl-2-picryl-hydrazyl (DPPH)
method (Brand-Williams, Cuvelier, & Berset, 1995). The mineral elements (Ca, Fe and Zn)
were measured using an atomic absorption spectrophotometer. In vitro digestion was
performed by the method originally developed by Miller, Schricker, Rasmussen & Van
Campen (1981), was modified by Bosscher, Lu, Cauwenbergh, Van Caillie-Bertrand,
Robberecht, & Deelstra (2001) and Jovaní, Viadel, Laparra, Barberá & Farré (2001). The
digested apple sample was used to determine the bioaccessibility of antioxidant capacity, iron,
zinc and calcium. Determination of resistant and digestible starch was used proposed by Goñi,
Garcia-Diz, Matas & Saura-Calixto (1996).

RESULTS & DISCUSSION

The antioxidant activity, mineral and starch content and bioaccessibility of apple samples were
significantly affected by the processing and digestion conditions. Therefore, these results
indicated that in-vitro digestion has a noticeable effect on the antioxidant concentration, IC50,
with much lower values (a smaller IC50 value corresponds to a higher antioxidant activity) of
apple samples compared with those untreated and non-digestion. Apple has the highest calcium
content (30.33±1.94 mg/100g), iron (14.46±3.49 mg/100g) and zinc (6.22±0.91 mg/100g).
High hydrostatic pressure increased the mineral contents availability by 2.11-303.00% for
calcium, 4.63-10.93% for iron and 8.68-28.93% for zinc. The dialyzability and solubility of
calcium, iron and zinc with respect to the values for the untreated sample were reduced by this
high pressure technique. Digestible starch content in the untreated sample was lowest than the
samples treated with high hydrostatic pressure but opposite effect was observed with resistant
starch.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1675
 Table 1: Effect of high pressure hydrostatic on DPPH free radical scavenging activity (IC50), mineral
elements and starch content in apple sample with non- and in-vitro digestion at treatment time different.
Apple samples
Untre ate d 500 MPa/2min 500 MPa/4min 500 MPa/8min 500 MPa/10min
IC 50 (mg/mL) 4.07±0.06a 4.81±0.42b 4.66±0.18b 4.58±0.02b 4.47±0.01bc
a a b b
IC 50 in-vitro digestion (mg/mL 0.28±0.04 0.20±0.06 0.04±0.03 0.08±0.07 0.09±0.06b
a b b b
Calcium solubility (%) 14.41±0,42 1.03±0.12 1.05±0.32 1.09±0.11 6.49±0.98c
b b b b
Calcium dialysis (%) 79.06±5.26 5.74±1.22 5.74±0.91 4.56±1.18 15.60±3.84c
a a b a
Iron solubility (%) 0.69±0.14 0.59±0.04 0.93±0.18 0.56±0.06 0.61±0.14a
a ab c b
Iron dialysis (%) 0.67±0.27 0.84±0.27 0.38±0.09 1.09±0.30 0.70±0.09a
ab b a a
Zinc solubility (%) 1.43±0.37 2.19±1.18 1.36±0.58 0.89±0.21 1.53±0.34ab
a b c
Zinc dialysis (%) 1.12±0.30 0.37±0.25 0.43±0.16b 6.38±0.99 4.29±0.49d
a a a b
Total starch (Holm`s method) 81.68±3.37 85.23±1.22 86.97±0.31 97.18±1.91 99.77±7.34b
Digestible starch (% D) 75.45±0.71a 76.58±1.73a 83.78±7.72a 96.43±7.00b 100.44±6.49b
Resistent starch (% R) 19.53±1.89a 19.42±4.47a 14.93±1.23ab 4.41±0.48c 3.12±0.51c
Total starch (% (D+R)) 95.07±3.00a 96.05±6.20a 98.68±8.95a 100.87±7.48a 103.52±6.99a
Different letters in the same row indicate that the values are significantly different (p < 0.05).

CONCLUSION
In summary, the antioxidant capacity, mineral and starch content of fruit may be
underestimated in the literature because the extraction solvents usually used do not allow a
complete release of antioxidant compounds. On the other hand, the analysis of in-vitro
digestive enzymatic extracts suggests that the antioxidant activity of fruit in the human gut may
be higher than what might be expected from literature data based on measurements of aqueous-
organic extracts. It is possible that changes to the tissue matrix induced by high hydrostatic
pressures, for example disruption of plant cell walls, resulted in the release of compounds with
antioxidant actions and increased mineral and starch content into the extracellular environment.
Consumption of apple under high hydrostatic pressure may supply substantial antioxidants,
mineral and starch which may provide health promoting and disease preventing effects.

REFERENCES
[1] Brand-Williams W., Cuvelier, M. E. & Berset, C. 1995. Use of a free radical method to evaluate
antioxidant activity. LWT- Food Science and Technology, 28, 25–30.
[2] Miller D. D., Schricker B. R., Rasmussen R. R., & Van Campen D. 1981. An in vitro method for
estimation of iron availability from meals. American Journal of Clinical Nutrition, 34, 2248-2256.
[3] Bosscher D., Lu Z., Cauwenbergh R.V., Van Caillie-Bertrand M., Robberecht H., & Deelstra H.
2001. A method for in vitro determination of calcium, iron and zinc availability from first-age infant
formula and human milk. International Journal of Food Sciences and Nutrition, 52, 173–182.
[4] Jovaní M., Viadel B., Laparra M., Barberá R., & Farré R. 2004. Improvement of analytical conditions
of mineral caco-2 cells uptake assays. Food Science and Technology International, 10, 197–201.
[5] Goñi I., Garcia-Diz L., Matas E., & Saura-Calixto F. 1996. Analysis of resistant starch: a method for
foods and food products. Food Chemistry, 56 (4), 445-449.

1676
 Microbiological stabilization of Aloe vera (Aloe barbadensis Miller) gel by high

hydrostatic pressure treatments

J.E. Reyes a, G. Tabilo-Munizaga a, M. Guanoquiza a, A. Vega-Galvez b, M. Mirandab and M. Pérez-Wonb

a
Food Engineering Department, University of Bio-Bio, PO Box 447, Chillán, Chile. (gtabilo@ubiobio.cl)
b
Food Engineering Department, University of la Serena, PO Box 559, La Serena, Chile.
(avega@userena.cl)

INTRODUCTION
In recent years, the use of Aloe vera in the formulation of food products has assumed an
important role, mainly due to recognition of their functional properties [1]. However, due to its
highly perishable nature, mainly from the microbiology point of view, aloe gel should be
processed in order to extend its shelf-life. For Aloe vera gel stabilization, it is often necessary
to apply thermal processing, which can produce irreversible modifications of the functional
components. A non-thermal technology such as high hydrostatic pressure (HHP) can be a good
alternative to conventionally thermal processing for aloe gel, because it has the potential to
inactivate spoilage microorganisms without significantly affecting the sensory, nutritional and
functional properties, and so increase the shelf-life [2]. Thus, the aim of this study was to
evaluate the application of HHP on microbial behavior and the shelf-life extension of the Aloe
vera gel during cold storage at 4 ºC.

MATERIALS & METHODS

The gel was obtained from whole fresh leaves of Aloe vera, which was homogenized twice by
a Warring-blender at high speed for 5 min, and then settle down for 24 h at 4oC. Gel samples
were packed in polyethylene flexible pouches and subjected to 300, 400 and 500 MPa for 1, 3
and 5 min at room temperature. HHP treatments were performed in a 2 litter processing system
(Avure Inc., Kent, WA, USA). Unpressurized (control) and pressurized samples were stored at
4°C for up to 90 days. All samples were analyzed at regular intervals for numbers of aerobic
mesophilic and psychrophilic microorganisms, Enterobacteriaceae and fungi by standard
methods. A re-parameterized version of the modified Gompertz equation [3] was used to
estimate the growth kinetic parameters, including the shelf-life. Data were analyzed by one-
way analysis of variance (ANOVA) and mean separations were obtained using the LSD test;
the level of significance was set a p < 0.05 level.

RESULTS & DISCUSSION

All HHP-treatment applied were able to significantly reduced (p < 0.05) counts of microbial
populations studied, except for mesophilic count in samples treated at 300 MPa/1 min, to non-
detectable levels (< 1.0 CFU/ml), reaching a reduction of ca. 2 to 4 log cycle (data not shown).
During storage at 4°C, the aerobic mesophilic and psychrophilic counts followed a similar
behavior (Figure 1A and 1B). In control samples, on the first day of storage showed mesophilic
counts higher than 2.0 log CFU/ml, which is considered the upper acceptable limit for Aloe
vera gel by WHO [4], and were found to be > 8.0 log UFC/ml at the end of storage period. In

11th International Congress on Engineering and Food - Athens, Greece, 2011 1677
contrast, in samples treated at 300 MPa (1, 3 and 5 min) and 400 MPa (1 min), this value was
reached after 18, 24, 35 and 53 days, respectively. In samples pressurized at 400 MPa (3 and 5
min) and 500 MPa (1, 3 and 5 min), the counts were kept below the detection limit during the
whole storage period. Enterobacteriaceae and fungi were not detected after any pressure
treatment and the number of survivors was kept below the detection limit during the whole
storage. A B
10 10

9 9

Microbial growth (log CFU/ml)

8
Microbial growth (logCFU/ml)

7 7

6 6

5 5

4 4

3 3

2 2

1 1

0 0
0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90

Storage time (days) Storage time (days)

Figure 1. Growth curve of (A) aerobic mesophilic and (B) psychrophilic on aloe gel storage at 4°C.
Control (z), 300 MPa/1 min (S), 300 MPA/3 min („), 300 MPa/5 min (T), 400 MPa/1 min (U), 400
MPa/3 min (…), 400 MPa/5 min (V), 500 MPa/1 min (¡), 500 MPa/3 min (²), 500 MPa/5 min (Ì).
Data were fitted by a re-parameterized version of the Gompertz equation. Bars denote standard deviation
of the mean.

On the other hand, the microbial shelf-life estimated for aloe stored a 4°C was between 18
and 53 days for the samples treated at 300 MPa (1, 2, and 3) and 400 MPa (1 min),
respectively. However, for the samples treated over 400 MPa for 3 min was > 90 days. The
long shelf-life of aloe gel treated by HHP-treated was attributed to the high initial microbial
reduction, as well as to increase in the duration of the lag phase (Ȝ) and a decrease in the
growth rate (μmax) of injured microorganisms (data not shown).

CONCLUSION
In this study, HHP-treatment at 400 MPa for 3 min and above was sufficient to produce
microbiology stable Aloe vera gel up to 90 days stored at 4°C.

ACKNOWLEDGMENTS: This study was financially supported by FONDECYT program

(project 1090228).

REFERENCES
[1] Miranda, M., Maureira, H., Rodríguez, K. &Vega-Gálvez, A. 2009. Influence of temperature on the
drying kinetics, physicochemical properties, and antioxidant capacity of Aloe vera (Aloe
Barbadensis Miller) gel. Journal of Food Engineering, 91(2), 297-304.
[2] Patterson, M.F. 2005. Microbiology of pressure-treated foods: a review. Journal of Applied
Microbiolgy. Food Science and Technology International, 98, 1400–1409.
[3] Corbo, M.R., Del Nobile, M.A. & Sinigaglia, M. 2006. A novel approach for calculating shelf life of
minimally processed vegetables. International Journal of Food Microbiology, 106, 69-73.
[4] WHO. 1999. Monographs on selected medicinal plants. Vol. 1., World Health Organization. Geneva,
Switzerland.

1678
 Establishment of a processing method for tofu using high pressure compared to the heat-

induced method

Yuri Jibua, Keiko Nakamuraa, Ai Teramotob, Hiroko Kuwadac, Michiko Fuchigamic

a
Department of Nutritional Science, Okayama Prefectural University, Soja, Japan (yjibu@ fhw.oka-
pu.ac.jp; keikotin@yahoo.co.jp)
b
Department of Health and Nutrition, Kanto Gakuin University, Yokohama, Japan (teramoto@kanto-
gakuin.ac.jp)
c
Department of Nutrition and Life Science, Fukuyama University, Fukuyama, Japan
(kuwada@fubac.fukuyama-u.ac.jp; fuchigam@fubac.fukuyama-u.ac.jp)

INTRODUCTION
Currently, the production method used for packed tofu (soybean curd) needs a lot of energy for
heating and cooling. The amount of added thermal energy is high and the environment of the
factory for tofu production is bad due to steam from heating. At high hydrostatic pressure,
proteins are denaturated and microorganisms are inactivated [1]. Extensive conformational
changes in protein are not caused below 300 ~ 400 MPa so denaturation only occurs at high
pressures [2]. Thus, high pressure is more useful than heating for food production, not only
food that contains protein but also jam and fruit sauce. Therefore, using high pressure for tofu
production to reduce thermal energy will be examined. Also, quality of high-pressure-induced
(HP-tofu) and heat-induced tofu (H-tofu) will be compared. The objectives of this study are to
establish a process for HP-tofu and compare it with H-tofu and heat-induced Market tofu (M-
tofu).

MATERIALS & METHODS

A mixture of soymilk (300 g) and 4 g or 6 g of 14.5% coagulant (Enden Nigari, Ako Kasei
Ltd.) solution was vacuum packed. Using a high pressure food processor (Kobe Steel Ltd.), it
was pressurized at 400, 500 and 600 MPa and kept at 25°C or 50°C for 10, 20, 30, 40 and 60
min (HP-tofu) [3]. The mixture was also heated for 30, 40 and 60 min at 88°C (H-tofu). The
tofu was then sliced into 1cm thick pieces using an ultrasonic sample cutter (Yamaden Ltd.).
After taking a photograph, the rupture stress and strain were measured by a creepmeter
(Rheoner, RE-33005, Yamaden Ltd.). The structure was observed with a cryo-scanning
electron microscope (S-4500, Hitachi Ltd.) and a sensory evaluation was done.

RESULTS & DISCUSSION

The rupture stress and strain of HP-tofu with 4 g and 6 g coagulant were compared in Figure 1
(including H-tofu). When the sample was pressurized at 25ÛC and 400 MPa for 60 min, it did
not coagulate. However, it coagulated above 500 MPa. As time of pressurization became
longer, the HP-tofu became firmer. As both pressure (from 500 MPa to 600 MPa) and
temperature (from 25ÛC to 50ÛC) increased, HP-tofu became more firm. Thus using a sensory
test, HP-tofu pressurized at 50ÛC and 500 MPa for 60 min, or at 50ÛC and 600 MPa for 30 min
was preferred as smoothest for mouthfeel and firmness and taste were preferred.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1679
It was found that HP-tofu, using the same amount of coagulant (4 g) as Market tofu, was softer
than M-tofu, but HP-tofu with 6 g of coagulant became harder. This HP-tofu was evaluated as
too hard and lacked smoothness. Also, H-tofu became spongy and the appearance became
worse when heating time was increased.
Coagulant 4g   Coagulant 6g

High-pressure-induced tofu Heat-induced tofu

400 MPa 500 MPa 600 MPa 0.1 MPa 400 MPa 500 MPa 600 MPa
150
150
25njC 25njC 88njC 25njC
Rupture stress (x10 N/m )

50njC
2

Rupture stress (x10 N/m )

2
50njC 50njC 50njC
2

2
100
100

50
50

0 0
60 30 40 60 10 20 30 30 40 60 60 20 30 40 60 10 20 30
60
25njC 25njC 60
50njC 88njC 25njC
50njC 50njC
50 50 50njC
Rupture strain (%)

Rupture strain (%)

40 40

30 30

20 20

10 10

0 0
60 30 40 60 10 20 30 30 40 60 60 20 30 40 60 10 20 30
Time (min) Time (min)
Figure 1. Rupture stress and strain of high-pressure-induced tofu (HP-tofu) and heat-induced tofu
(H-tofu) made of coagulant (4g or 6g) and soymilk (300g).

CONCLUSION
Tofu with 4 g of coagulant pressurized at 50ÛC for 60 min at 500 MPa or for 30 min at 600
MPa was preferred by a sensory test. Although this tofu was softer than Market tofu, the taste
was good due to smooth texture, because there were smaller air bubbles and the gel network of
protein was homogeneous.

REFERENCES
[1] Hayashi, R. 1989. Use of High Pressure in Food. San’ei Press, Kyoto.
[2] Hayashi, R. & Balny, C. (1995). High Pressure Bioscience and Biotechnology. Elsevier, Amsterdam.
[3] Kato, H.N., Teramoto, A., & Fuchigami, M. 1997. Pectic Substance Degradation and Texture of
Carrots as Affected by Pressurization. Journal of Food Science, 62(2), 359-362, 398.

1680
 Enhanced Infusion under High Pressure: New Insights

Swetha Mahadevan and Mukund V. Karwe*

Department of Food Science, Rutgers University, New Brunswick, NJ 08901, USA

*(karwe@aesop.rutgers.edu)

INTRODUCTION
Osmotic dehydration has been traditionally used in the food industry for infusion of substances
into food materials. Diffusion, the underlying phenomenon of osmotic dehydration, is
inherently a slow process and there is a need for additional methods that can accelerate mass
transport rates. High hydrostatic pressure processing (HHPP) is one such alternative. Studies
have shown that HHPP can enhance infusion of substances, such as salt and sugar, into fruits,
vegetables and meat [1], [2], [3]. The commonly accepted explanation is that upon exposure to
HHPP, the cell membranes of the food tissues rupture thereby decreasing the resistance to
infusion [2]. This explanation, however, is only causal and other mechanisms that increase
driving force or reduce resistance to infusion even without changes in the microstructure may
be operative, resulting in enhanced infusion. This study was undertaken to test the hypothesis
that other possible mechanisms exist that can enhance infusion under high pressure. The model
system of study chosen was an antioxidant-fruit system of quercetin- frozen-thawed cranberries
where quercetin was the infusant and frozen-thawed cranberry was the fruit substrate. We find
that enhanced infusion of quercetin into frozen-thawed cranberries occurred without any
additional cell-membrane permeabilization under high pressure indicating other possible
mechanisms at play during high-pressure induced infusion.

MATERIALS & METHODS

Scarified-frozen-thawed cranberries were immersed in a solution of glycerin-ethanol (1:1)
carrying 0.5% quercetin and high-pressure processed in bags at pressures between 100 and 551
MPa for 1-60 minutes at room temperature; cranberries had to be scarified since diffusion
through intact cranberry skin is negligible even under high pressure. Amount of quercetin
infused in the processed samples was measured by RP-HPLC. Cell-membrane
permeabilization changes were quantified using an electrophysical measurement based on
impedance spectroscopy [4]. The cell-membrane permeabilization changes were expressed in
terms of a permeabilization index, Zp. Fruit microstructure was analyzed using light
microscopy. All measurements were made in triplicates.

RESULTS & DISCUSSION

Preliminary experiments had shown that diffusion through cranberry skin was almost
negligible even under high pressure. In order to reduce this mass transfer barrier, fruit surface
was partially scarified by making sub-millimeter pinholes.
High pressure treated cranberries showed significantly higher levels of quercetin compared to
control. The amount of quercetin infused in cranberries processed under high pressure was
found to be three times that in cranberries processed at ambient conditions (control). In
addition, infusion under high pressure was much faster than that at ambient conditions: the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1681
amount of quercetin that was infused at ambient conditions in 3 hours was infused in only 10
min under high pressure (Figure 1). Furthermore, the amount of quercetin infused into
cranberries processed under high pressure was found to be independent of the applied pressure
in the range investigated. This observation was unlike other studies, which have shown
increase in diffusion with increase in applied pressure [2,3].
The cell-membrane permeabilization changes due to HHPP were recorded and expressed in
terms of Zp (permeabilization index). There was no significant difference in cell-membrane
permeabilization between control and
HHPP treated cranberries. Unlike other
studies [1], however, we did not find
additional cell permeabilization in frozen-
thawed cranberries after HHPP. These
results were in agreement with our
observations of fruit tissue microstructure
obtained by light microscopy. Our results
suggest that enhanced infusion in HHPP
may be caused by factors other than cell-
membrane permeabilization and that more
studies are required to understand the Figure 1. Variation of quercetin infused with applied
fundamental mechanisms of infusion under pressure in frozen-scarified-thawed cranberries
HHPP.

CONCLUSIONS
In scarified-frozen-thawed cranberries, HHPP treated cranberries showed a significant increase
in the amount of quercetin infused over that in those treated at ambient conditions. Under
HHPP, mass transfer rates were enhanced and processing time was shortened. Furthermore, the
amount infused into cranberries under high pressure was found to be independent of the
applied pressure in the range investigated. Remarkably, HHPP had no additional effect on cell-
membrane permeabilization in frozen-thawed cranberries. Our results of similar degree of
permeabilization in all samples and observed enhanced infusion suggest that cell-membrane
permeabilization may not be the only cause for high pressure induced infusion. Further studies
to investigate other mechanisms operating during high pressure induced infusion are required.
This work elucidates important aspects of the science of pressure-enhanced infusion. In
addition, this work demonstrates the potential of HHPP to develop nutraceutical-enriched food
products.

REFERENCES
[1] Rastogi N.K., Angersbach A., Niranjan K., and Knorr D. 2000. Synergistic effect of high hydrostatic
pressure pretreatment and osmotic stress on mass transfer during osmotic dehydration. Journal of Food
Engineering, 45, 25-31. [2] Sopanangkul A., Ledward D.A., and Niranjan K. 2002. Mass transfer during
sucrose infusion into potatoes under high pressure. Journal of Food Science, 67(6), 2217-2220. [3]
Villacis M.F., Rastogi N.K., Balasubramanian V.M. 2008. Effect of high pressure on moisture and NaCl
diffusion into turkey breast. LWT, 41, 836-844. [4] Angersbach A., Heinz V., and Knorr D. 1999.
Electrophysical model of intact and processed plant tissues: cell disintegration criteria. Biotechnology
Progress. 15, 753-762.

1682
 Structural changes of pectin methylesterase from orange peel subjected to thermal and

high pressure processing

Z.Alexandrakisa, T. Papadopoulosb, Ph.Stavrosb, G.Katsarosa, P.Katapodisa, G.Nounesisb, P.Taoukisa

a
Laboratory of Food Chemistry and Technology, School of Chemical Engineering,National Technical
University of Athens, Athens, Greece, taoukis@chemeng.ntua.gr
b
Biomolecular Physics Laboratory, IRRP, National Centre for Scientific Research Demokritos,Aghia
Paraskevi, Greece, nounesis@rrp.demokritos.gr

INTRODUCTION
Pectinmethylesterase (PME), an endogenous enzyme found in fruits and vegetables, affects the
quality characteristics of final products (such as juices). In the literature there is a significant
number of papers describing the effect of high pressure (HP) and temperature on PME activity
from different fruits and vegetables, such as orange, tomato, peach, strawberry, green beans
and papaya. The results found in literature and the obtained results from experiments
conducted from our laboratory show that the inactivation depends on the origin of the enzyme
[1,3]. Despite the extensively studied PME inactivation using thermal and HP processing, there
is limited work done on the description of structural changes of this enzyme processed by HP.
The objective of this research was to study the structural changes of PME before and after
treatment of the enzyme at temperatures (40-80oC), combined with pressures (0.1-800 MPa).

MATERIALS & METHODS

The effect of high pressure (HP) in conjunction with mild temperature on the activity and the
conformation of PME purified from orange peel (Valencia var.) by cation-exchange
(UNOspere S, Biorad) and gel filtration (Biogel P-30, Biorad) chromatography was studied.
High pressure treatments were achieved using a pilot scale HP equipment with a maximum
operating pressure and temperature of 1000 MPa and 100°C respectively (Food Pressure Unit
FPU 1.01, Resato International BV, Roden, Holland). The secondary and tertiary structure of
HP treated and untreated purified PME was analyzed using a Jasco-715 spectropolarimeter in
the near and far ultraviolet regions. The assay for the measurement of PME activity
spectrophotometrically at 620 nm was described by Cameron [2].

RESULTS & DISCUSSION

The far –UV CD characterization of HP treated and native PME is depicted in Fig.1. The far
UV CD spectra of all samples were analyzed immediately after HP treatment. A characteristic
for ȕ-structure (ȕ-sheets and turns) proteins negative peak at ~ 218 nm was observed for PME.
HP processing resulted in simultaneous inactivation of PME and decrease of the ȕ-structure
fraction of their secondary structure.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1683
 PME_untreated
PME_200 MPa_40oC,40 min, R.A. = 0,74
10 PME_600 MPa_40oC, 7 min, R.A. = 0,39
[T]*10 (deg.cm .dmol ) PME_ 0,1 MPa_60oC, 5 min, R.A. = 0,76
-1

0
2

-10
-3

-20

-30
200 220 240 260
Wavelength (nm)

Figure 1. Far-UV CD spectra of untreated and HP treated PME at 25o C.

CONCLUSION
The secondary structure of PME treated by HP changed as evidenced by CD analysis. The
intensity of negative peak in the CD spectra as well as the relative residual activity of PME
decreased with the increase of pressure and treatment time. The obtained results explain the
reduced enzyme activity (inactivation) of this enzyme after processing at certain conditions
which is in close agreement with its conformation change.

ACKNOWLEDGEMENTS
This research has been co-financed by the European Union (European Social Fund – ESF) and
Greek national funds through the Operational Program "Education and Lifelong Learning" of
the National Strategic Reference Framework (NSRF) - Research Funding Program: Heracleitus
II. Investing in knowledge society through the European Social Fund.

REFERENCES
[1] Balogh T., Smout C., Ly Nguyen B., Van Loey A.M., Hendrickx M.E. 2004. Thermal ana High-
Pressure inactivation of carrot pectinmethylesterase : from model system to real foods, Innovative
Food Science and Emerging Technologies, 5, 429-436.
[2] Cameron, R. G.;,Buslig, B. S.,Shaw, P. E.1992. Adaptation of a spectrophotometric assay for
pectinmethylesterase to a kinetic microplate reader. J. Food Sci., 57, 1006-1008.
[3] Nienaber U. and Shellhammer T.H.2001.High-pressure processing of orange juice : kinetics of
pectinmethylesterase inactivation, Journal of Food Science, 66 (2), 328-331.

1684
 Innovative value propositions for the food industry through non-thermal processing

techniques

Francisco Purroy a, Carole Tonelloa

a
NC Hyperbaric SA, Burgos, Spain (f.purroy@nchyperbaric.es )

INTRODUCTION
Emerging, Non-Thermal processing technologies are becoming widespread in the food
industry, mainly as post-packaging interventions for food safety assurance and stabilisation of
All-Natural, preservative-free propositions. Techniques such as High Pressure Processing
(HPP) prove to be effective and economically feasible, showing now consistent double-digit
growths. Being increasingly implemented in the last decade, solutions like HPP are opening
other options to the food industry worldwide: developments in the last 24 months include both
application or product innovations, and new value-added industrial processing scenarios for the
food sector. This paper reviews some of these advances including: substitution of traditional
thermal techniques for novel product manufacturing; development of new functional
beverages; tenderisation of low value meat cuts; and the new value proposals being offered by
copackers and refrigerated services suppliers.

MATERIALS & METHODS

This paper reviews activities by various organisations including research bodies such as
CSIRO-Food Science Australia and DIL (German Institute of food technologies) in Germany;
institutions such as MLA (Meat&Livestock Commission) in Australia; and companies such as
Fonterra Cooperative Group or New Image Natural Health in New Zealand, or Cargill and
Hormel Foods, in the United States. Their work has resulted in published whitepapers (DIL,
CSIRO), patents (Hormel Foods, Fonterra) and commercial potentialities through the use of
high pressure processing techniques (MLA, Cargill, Hormel Foods, New Image Natural
Health).

RESULTS & DISCUSSION

Substitution of thermal treatments and development of new functional products
Extensive research carried out in Germany, New Zealand and the US over the last years led to
the creation of new opportunities that are being adopted by food industries, including: a)
substitution of traditional thermal treatments in the manufacturing of spreadable sausages, liver
sausages and cooked-cured loin products. High pressure processing techniques developed in
Germany are allowing manufacturers to eliminate heat steps in their process flow, in order to
improve textures or achieve protein gelification and firmness. The high isostatic pressure step
also offers dietary and sensory advantages, as it retains valuable micro-molecules such as
aroma components and vitamins, which bonds can not be modified by HPP process. b) creation
of new opportunites in the functional products space. Private company research from a major
dairy multinational in New Zealand creates a market space for probiotic, functional juices in

11th International Congress on Engineering and Food - Athens, Greece, 2011 1685
which no culture management is necessary and a extended shelf-life is achieved, too. HPP
seems to offer a solution in probiotic propositions, in which a thermal pasteurisation is not an
option as heat inactivates the probiotic strains.
Tenderisation of red meats
Another research field showing now strong commercial perspectives has been developed by
scientists mainly in Australia and North America. By minimally processing low value red meat
cuts with usage of high pressure processing principles, with parameters around 200MPa-
2000bar for a few minutes (in opposition to the usual 600MPa-6000bar used for post-
packaging high pressure pasteurisation) consistent results are shown including higher water
binding in meat proteins, improved tenderness, decreased collagen taste and better overall
sensorial results. This is giving very interesting possibilities to industries willing to add value
to low value beef and lamb cuts. MLA (Meat and Livestock Australia) is aiming to take the
project into industrialisation phase.
Hormel Foods, contemporaneously to the abovementioned research, has obtained in 2010 a
patent for inhibition of post-mortem glycolisis by the use of high pressure processing.
Processing meat cuts of pork, turkey, beef under pressures of a minimum 175MPa and
maximum 320MPa for a few seconds or minutes in a right-after slaughtering intervention,
inhibition of rigor mortis is achieved in the muscles, rendering improved juiciness, texture and
palatability of meats.
Value-adding refrigerated services and Co-packing
Toll processing of innovative technologies is a scheme that has been increasingly utilised in the
food industry over the last five years. At the turn of the decade, innovative value services
appear, now not only offered by copackers but also by companies dedicated to serve food
industries as logistic platform and integral refrigerated services solution. Such companies are
strategically moving towards adding more value to their customers: now they target not only
stock management, chilled supply chain or JIT deliveries. In a new business case, organisations
offering refrigerated services are currently adding more value to their service, by offering new
technological capabilities and granting customers the access to post-packaging techniques for
shelf life extension or pathogen destruction. Thus food&beverage industries who are shipping
their product to these cold storage facilities, can also benefit from other innovative processing
solutions they don’t have in their own facilities.

CONCLUSION
High Pressure Processing (HPP) technology seems to keep on broadening its spectrum of
potentialities and commercial value propositions for food industries, and proves that non-
thermal technologies can prove beneficial and profitable in various fields and sectors of
application.

REFERENCES
[1] Jung C, Tonello C et al,. 2011. High Pressure Processing of Foods. Alternatives to conventional food
processing, 6, 254-306
[2] Sikes A et al, 2010. A proposed mechanism of tenderising post-rigor beef using high pressure
treatment. Meat Science 8390-399.
[3] Purroy F et al, 2011. Value adding refrigerated services through nonthermal processing techniques.
International Meat Tech sessions, Chicago, USA. 13-16 April, 2011.

1686
 Fractionation of liquid egg yolk:

Influence of chemical and structural characteristics of egg yolk granular and plasma

fraction on the continuous centrifugal separation process

Strixner Thomas, Michael Betz, Ulrich Kulozik

Chair for Food Process Engineering and Dairy Technology,

ZIEL Food and Nutrition Research Center, Technische Universität München, Weihenstephaner Berg 1,
85354 Freising-Weihenstephan, Germany
thomas.Strixner@wzw.tum.de

INTRODUCTION
Protein stabilised emulsions play a major role in many areas of the food industry. One of the
most complex protein-based emulsifier systems is hen’s egg yolk. The emulsifying properties
of the two main egg yolk fractions, egg yolk granules (HDL) and plasma (LDL), are known to
be completely different for each fraction. Plasma contains mainly low-density lipoproteins
(LDL) and globular glycoproteins. LDL, whith apoproteins called lipovitellenins, is the main
constituent of egg yolk. Granules contain phosvitin, a phosphoprotein, and high-density
lipoprotein (HDL). In addition, small amounts of LDL can be found in insoluble granule
aggregates. The complex between HDL and phosvitin is held together by phosphocalcic
bridges.
Changes in pH or salt concentration influence the chemical and structural characteristics for
each fraction in a different way. Recent studies substantiate that a separate use of each fraction
in innovative food products will offer new ways in emulsification technology. However, so far
no systematic study deals with the separation process as a function of physico-chemical factors.
This study applies a wide range of environmental conditions and centrifugal separation
parameters enabling the determination of the optimal operating conditions for the
centrifugation of liquid egg yolk, which is a highly complex system. A pilot scale bowl
centrifuge is used to evaluate the process parameters varying centrifugal acceleration and
residence time. The influences of different pH and salt concentrations, where granules are in
their native and a disintegrated form, are investigated. Calcium is added in different
concentrations to influence the phosphocalcic bridges within the HDL complexes.

MATERIALS & METHODS

The maximum of depositing granula at physiological milieu conditions was assessed by a
modified method of McBee and Cotterill [1]. The egg yolk was at first diluted (1:2, w/w) in an
isotonic sodium chloride solution (0.15 M NaCl) and stirred gently for 1 h before
centrifugation at 10.000g for 45 min at 10 °C.
The dry matter DM of the egg yolk solution, the plasma and granula fraction was determined
by the sea sand method according to handbook of analysis methods, volume 6 C, 35.3,
VDLUFA, 1985.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1687
Rheological measurements were carried out with a controlled shear rate rheometer AR 1000
from TA Instruments (Eschborn, Germany) equipped with cone plate geometry (d=6 cm;
angle: 2°, gap width 400 μm)

RESULTS & DISCUSSION

It became evident that the granula fraction represents no homogeneous component, but consists
of different subclasses with different size, density and structure. This inhomogeneity leads to a
subclass depending sedimentation behaviour of the whole granula fraction in egg yolk. A
variation within the egg yolk pH to acidic milieu conditions results in changes of each subclass
constitution. Especially sub fractions with small particle size and high protein content
incorporate a higher amount of granular LDL. Therefore, the particle density is decreased,
which leads to a much lower sedimentation velocity. Based on these findings, the impact of
variable rheological properties on the sedimentation behaviour of polydisperse colloidal
suspensions with high particle concentrations is discussed. It was detected that if a limiting
particle concentration is reached all particles within the polydisperse particle collective are
sedimenting with the same velocity. At the so called zone-sedimentation the high fluid
viscosity as well as strong particle-particle interactions are resulting in constrained separation
effects.
100

22% DM 50°C
separation efficiency K[%]

22% DM 4°C
80

60

34% DM 4°C

40

20

Granula
0
0 2000 4000 6000 8000 10000 12000

g-force
Figure 1. Separation efficiency Ș of the egg yolk granula fraction in dependency of the g-force for
different dry matter concentrations in the egg yolk solution (22 % and 34 %) and product temperatures
(4°C and 50°C).

CONCLUSION
The study offers a detailed description for the separation of egg yolk granula and plasma
fraction under varying parameters in a laboratory scale. Additionally, the results allow a deeper
insight in the structural complexity of the LDL and HDL egg yolk fraction and offer therefore
new information for the techno-functional properties of egg yolk fractions.

REFERENCES
[1] McBee, L. E., Cotterill, O. J (1979). Ion exchange chromatography and electrophoresis of egg yolk.
Journal of Food Science, 44, 656–660.

1688
 Refining of crude canola oil using PSA ultrafilteration membrane

Ali Rafea, Seyed Mohammad Ali Razavib, M. H. Haddad Khodaparast a,b

a
Department of Food Science and Technology, Ferdowsi University of Mashhad (FUM),
PO Box: 91775-1163, Mashhad, Iran (alirafe1400@yahoo.com)
b
Department of Food Science and Technology, Ferdowsi University of Mashhad (FUM),
PO Box: 91775-1163, Mashhad, Iran (s.m.a.razavi@um.ac.ir)

ABSTRACT
The aim of this paper was to study the behavior of spiral wound configuration of polysulfone
amide (PSA) membrane with 20 kDa molecular weight cut off (MWCO) in pilot-plant scale
equipment used for refining micella canola oil under different operating conditions. Refining
process parameters including phospholipids (PLs), color and free fatty acids (FFAs) were
determined to express retentions (R%) and ultrafiltration process performance parameters such
as permeate flux, membrane fouling and hydraulic resistances were determined. The results
showed that the permeate flux was decreased considerably with increasing process time,
although it was increased by increasing temperature from 30 to 50ÛC and transmembrane
pressure from 1.5 to 2 bar. The irreversible fouling resistance (Rif) and percentage of fouling
were decreased as the temperature or transmembrane pressure increased. The results showed
that the concentration polarization resistance (Rrf) was much higher than other resistances;
therefore reversible resistance had an important role in total hydraulic resistance. In this paper,
the retention of PLs, FFAs and color was so interesting due to improving the efficiency of oil
refining process. The percentage of retention of PLs and FFAs was increased by temperature
and decreased by transmembrane pressure and time; however, there was no significant
difference in removing color under different operating conditions.

CONCLUSION
This study showed that membrane separation is effective in removing PLs, FFAs and
increasing color quality particularly removing chlorophyll pigment in canola oil. Despite the
promising results reported here, there is still a lot of work to be undertaken and phenomena
responsible for the selectivity of the separation, mainly. For example these trials can be used
for PES membrane which has the best efficiency for refining canola oil.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1689
1690
 Optimization of proteins recovery process from cheese whey

Cuellas Anahía, Jagus Rosab, Wagner Jorge R.a,c

a
Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Buenos Aires, Argentina
(acuellas@unq.edu.ar, jwagner@unq.edu.ar)
b
Facultad de Ingeniería, U.B.A., Buenos Aires, Argentina (rjagus@di.fcen.uba.ar)
c
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)

INTRODUCTION
Cheese whey, the main by-product in the manufacture of cheese, has a severe problem because of its
high biological oxygen demand (35-60 g/l). Hence, there is an increasing need to develop methods
for making use cheese whey, a nutritional by product, in order to avoid the negative environmental
implications and to give a better economic return. The most important cost-effective utilizations of
cheese whey are valorization technologies, such as recovery of protein and lactose from whey [1].
The aim of this work was to optimize the process to obtain a protein-containing ingredient from the
remaining whey of Argentinean “Cuatriolo” cheese production. In order to optimize the protein
recovery, physical and chemical treatments were investigated.

MATERIALS & METHODS

Fat removal from cheese whey was carried out by filtration and centrifugation. Two protocols were
assayed, Protocol I: heat whey until boiling, adjusting to pH 4.6 with acetic acid and keep the
heating to 90 ºC for 30 min. Protocol II: heat whey until boiling, holding at 90 ºC for 30 min and
adjusting to pH 4.6. Protein aggregates and supernatant were separated by centrifugation (Samples I
and II). The influence of fat presence and CaCl2 addition were evaluated on protocol II. Sample II
Ca: addition of CaCl2 (200 mg/ml) in the acidification step. Sample II fat: idem sample II without
fat elimination. Sample II Ca-fat: idem sample II Ca without fat elimination. Protein determination
was performed by the Lowry method. Yield was determined from the relationship between
precipitated protein and whey protein. Water retention capacity was determined from the ratio of
precipitate wet weight and precipitated proteins. Particle size distribution, expressed in % number,
was determined by laser scattering using a Malvern Mastersizer 2000E (Malvern Inst. Ltd, UK).

RESULTS & DISCUSSION

Comparison between samples I and II showed that a heat denaturation process previous to acetic
acid addition (protocol II), allows to obtain a better protein recovery (Figure 1a) with a higher
water retention capacity of aggregates (Figure 1b). Since whey proteins are relatively low
molecular weight and soluble at its isoelectric point, a thermal treatment is necessary to precipitate
them [2]. During this process, ȕ-lactoglobulin undergoes a structural change that exposes the S-S
groups, which play a central role in the formation of covalent "bridges" with other proteins. The
presence of CaCl2 and/or fat in protocol II did not enhance the protein recovery (Figure 1a). The
unfolding of the native structure of proteins, promotes the establishment of hydrophobic interactions
and disulfide bonds with protein molecules, carbohydrates and lipids. According to the results
observed, the presence of fat interferes with the formation of protein aggregates, resulting in a lower
yield.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1691
 80 a 50 b

Water retention capacity (g/g)

70

60 40

Yield (%)
50
30
40

30 20

20
10
10

0 0
I II II Ca II fat II Ca-fat I II II Ca II fat II Ca-fat
Sample Sample

Figure 1. Yield (a) and Water retention capacity (b) of protein aggregates

On the other hand, a decrease of water retention was observed in the presence of calcium (sample II
Ca, Figure 1b). Calcium is not involved in denaturalization process, but it takes part in the
aggregation of denatured proteins, intervening in the structure of the obtained aggregates. The
simultaneous presence of fat and CaCl2 (sample II Ca-fat) increase the amount of water retained in
the aggregated protein (47.62 g precipitate wet /g protein precipitate), nevertheless the alone
presence of fat does not modify the water retention. The yield of protein recovery is related to the
particle size of the aggregates. An increase in the particle size produces an increase in the recovery
of proteins. Protocol I yields particles smaller than protocol II, therefore lower protein recovery. A
higher particle size could be a consequence of a higher hydration degree of particles, which was
clearly observed in the water retention capacity of samples I and II (9.4 and 41.7 %, respectively).
Electron micrographs of aggregates (data not shown) allow to see that sample I is composed of
aggregates with enclosed structure. This spatial arrangement of aggregates could explain the low
water retention of the protein precipitate with protocol I. On the other hand, the protein aggregates
obtained from protocol II have a net-like open structure, capable of occluding water. The presence
of calcium and to a greater extent the presence of fat, show a lower yield of recovery (72.8 and 69.4
%, respectively) and this correlates with a smaller particle size.

CONCLUSION
The results showed that the highest yield of the process and an increase in water retention are
achieved when the acid is added after whey protein denaturalization at 90 ° C for 30 minutes. The
simultaneous presence of fat and CaCl2 increase the amount of water retention capacity Therefore,
the process of aggregation in whey protein should take into account both the design of suitable
protocol such as the presence of fat and additives. Hence, the optimization of the aggregation
process allows to reduce the volume of effluent from the cheese industry and getting new protein
ingredients for use in the food industry.

REFERENCES
[1] Smithers, G.W. 2008. Whey and whey proteins – From ‘gutter - to – gold’. Int. Dairy Journal, 18:
695-704.
[2] Hill. A. R., Irvine, D. M. & Bullock. D. H. 1982. “Precipitation and Recovery of Whey Proteins: A
Review”. Can. Inst. Food Sci. Technol. J. 15(3):155-160.

1692
 Production of adsorbents based on food waste (corn cobs) for removal of phenylalanine

and tyrosine from aqueous solutions

Adriana S. Francaa,b, Cibele C. O. Alvesb, Leandro S. Oliveiraa,b

a
Departamento de Engenharia Mecânica, Universidade Federal de Minas Gerais (UFMG), Av. Antônio
Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil (adriana@demec.ufmg.br)
b
Programa de Pós-Graduação em Ciência de Alimentos, UFMG, Belo Horizonte, MG, Brazil

INTRODUCTION
Phenylketonuria (PKU) and tyrosinemia (TYR) are hereditary disorders, in which the
metabolisation of the corresponding amino acids phenylalanine (Phe) and tyrosine (Tyr) is
impaired due to the deficiency of the respective enzymes, resulting in several problems in
untreated patients. PKU and TYR nutritional therapies are accomplished by rigorous diets
based on protein substitutes, either mixtures of free amino acids or protein hydrolysates. In
Brazil, such mixtures of free amino acids are imported and available at high costs. One
alternative to reduce costs is the use of residues from the food industry in the development of
protein hydrolysate in association to the use of low-cost adsorbents for Phe removal. Thus, the
objective of this work was to evaluate the performance of corn cobs in the production of
adsorbents for Phe and Tyr removal from aqueous solutions.

MATERIALS & METHODS

The adsorbent was prepared by thermo-chemical activation of corn cobs with H3PO4
(impregnation ratio of 85 % - 3 min at 25 °C followed by 1h carbonization at 400°C). Batch
experiments of adsorption were performed and the effect of contact time was evaluated at time
periods ranging from 5 min to 12 hours, for initial solution pH of 6 and adsorbent
concentration of 10 g L-1. Phe and Tyr concentrations was determined by a UV-Vis
spectrophotometer (Hitachi U-2010) at 257 and 273 nm, respectively. For evaluation of the
binary systems, batch adsorption tests were conducted employing equal volumes of each
aminoacid solution at the following concentrations: a) Phe-Tyr: Phe 500 mg.L-1; Tyr ranging
from 20 to 150 mg.L-1; b) Tyr-Phe: Tyr 50 mg.L-1; Phe ranging from 200 to 1500 mg.L-1.

RESULTS & DISCUSSION

The adsorption isotherms obtained for each aminoacid, in single and binary systems, are shown
in Figure 1. The shapes of all curves indicate favorable adsorption [1]. A comparison of the
curves obtained for single and binary adsorption of each aminoacid shows that, in the case of
Phe, the presence of Tyr favoured faster adsorption whereas the opposite behavior was
observed for Tyr. Such results indicate that in the binary systems Phe will be adsorbed
preferably in comparison to Tyr. The adsorption dynamics of the aminoacid mixture was also
evaluated using qe'/qe ratios, were the prime denotes the presence of another aminoacid [2]. In
general, three possible types of behavior are exhibited: qe'/qe > 1, indicating synergism (the
effect of the mixture is greater than that of the individual adsorbates in the mixture); qe'/qe < 1,

11th International Congress on Engineering and Food - Athens, Greece, 2011 1693
corresponding to antagonism (the effect of the mixture is less than that of each of the individual
adsorbates in the mixture); and qe'/qe = 1, non-interaction (the mixture has no effect on the
adsorption of each of the adsorbates in the mixture). The qe'/qe ratio was 1.14 for Phe
adsorption in the presence of Tyr, suggesting a slightly sinergistic effect. However, the qe'/qe
ratio for Tyr adsorption in the presence of Phe was 0.5, indicating that Tyr adsorption was
depressed by Phe in the binary solution.

60 10

50 8
40
6

qe (mg/g)
qe (mg/g)

30
4
20

10 2

0 0
0 100 200 300 400 500 0 5 10 15 20

Ce (mg/L) Ce (mg/L)

(a) (b)

Figure 1. Adsorption isotherms for (a) phenylalanine and (b) tyrosine removal at 25 °C ( pure; x
binary solution; solid lines correspond to Langmuir and LCM fits for single and binary systems,
respectively)

CONCLUSION
The feasibility of employing corn cobs as raw material in the production of adsorbents
for removal of phenylalanine and tyrosine from aqueous solutions was investigated.
The obtained results showed that Phe will be adsorbed preferably in comparison to
Tyr in a binary system and indicate that the produced adsorbent is a promising
alternative for reducing costs in the production of Phe-depleted protein hydrolysates.

ACKNOWLEDGEMENTS
The authors acknowledge financial support from the following Brazilian Government
Agencies: CNPq and FAPEMIG.

REFERENCES
[1] Do, D. D. 1998. Adsorption Analysis: Equilibria and Kinetics. Series on Chemical Engineering:
Australia. 2, 913p.
[2] Mahamadi, C.; Nharingo, T. 2010. Competitive adsorption of Pb2+, Cd2+ and Zn2+ ions onto
Eichhornia crassipes in binary and ternary systems. Bioresource Technology, 101, 859–864.

1694
 The effect of applied conditions on whey separation and fractionation using ultra- and

nanofiltration

Petra Zidova, Andrea Hinkova, Vladimir Pour, Zdenek Bubnik, Svatopluk Henke, Alena Salova and
Pavel Kadlec

Institute of Chemical Technology Prague, Department of Carbohydrate Chemistry and Technology,

Technicka 5, 166 28 Prague 6, Czech Republic (e-mail: petra.zidova@vscht.cz)

INTRODUCTION
Membrane separation processes find their applications in dairy industry since the beginning of
1970s and their number and importance is still rising. Nowadays, membrane separations form the
integral part of the dairy technology.
Whey is a liquid part separated from the curd during cheese production. Whey composition depends
on milk properties and technology used. The dry matter varies between 5.5 and 6.5 % (lactose, whey
proteins, minerals, non-protein nitrogen compounds, fats and acids) [1].
Utilisation of minor whey components represents a big challenge for whey processing also in the
future [2]. Membrane processes might be very effective in isolation of bioactive peptides, growth
factors or oligosaccharides. Ultrafiltration together with nanofiltration can be used for peptide and
amino acid fractionation, where not only the size of molecule but also its effective charge can
improve the NF separation [3].

MATERIALS & METHODS

Ultrafiltration module used: TIA (Bollene, France) equipped with inorganic tubular membranes
Membralox (Pall, France). Nanofiltration module used: ARNO 600 (Mikropur, Czech Republic).
NF membranes tested: Desal DL (GE Osmonics), NF-245 (Dow Chemicals) and TFC – SR100
(Koch).
Two types of solution were processed by UF followed by NF: dried sweet whey solution and fresh
natural whey.
All filtrations were performed in a retentate recycling mode at constant temperature and pressure.
Ultrafiltrations were carried out for purifying of solutions as single or multiple stages. The pH of
solutions was modified only before NF. During filtration the kinetics parameters were measured
followed by the measurements of apparent rejection Ri [%] of lactose, ions and whey proteins.

RESULTS & DISCUSSION

The filtration kinetics and separation efficiency of ultrafiltration and nanofiltration
It was carried out 5 series of ultrafiltrations experiments. Achieved mass concentration factors were
between 2.3 to 7 proving sufficient concentration of initial feed. The average permeate fluxes show
big differences. The 5 kD ultrafiltration was the slowest UF giving the fluxes of 14 - 19 l/h.m2.bar
for natural whey.
Pure water fluxes before filtration were very variable and illustrate extensive membrane fouling and
unsatisfactory cleaning. Cleaning procedure used sodium hydroxide, nitric acid and finally sodium
hypochlorite.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1695
Also the separation efficiency has been studied and expressed as Rejection. Results show that 100
and 500nm UF membranes were highly permeable for lactose molecules and lactose rejection on
5kDa membrane was only about 0.1 %, which documents minimum losses of lactose in retentate.
The rejection of whey proteins were measured during multiple step ultrafiltrations. The highest
protein rejection (84 – 100 %) was observed on 5 kDa membrane, 500nm membrane protein
rejection varied between 60 and 100 % and the one on 100nm membrane was 63 – 90 %. Using 5
kDa membrane will totally remove all protein which can be further used for production of protein
isolates or concentrates, however it reduces the velocity of whey purification due to the low
permeate flux on this membrane.
During NF tests especially the effect of pH on separation was observed. The average fluxes for
natural whey were 34 - 50 l/h.m2 on TFC-SR100, 63 – 68 l/h.m2 on NF-245 and 15 – 40 l/h.m2 on
Desal DL membrane. MCF values varied between 1 and 1.9 which is not sufficient for industrial
application. This was mostly caused by small membrane area and verification on larger module (e.g.
spiral wound) will be necessary.
The separation efficiency was significantly affected by pH of entering whey. The data confirm
increasing rejection of monovalent ions on the membrane TFC-SR100 at pH 6 and 5.5 with minimal
rejection at pH 5. Similarly, the rejection on lactose on this membrane reached the maximum at pH
6.5 and minimum at pH 5.5, however the differences in minimum and maximum lactose rejection
were only 2.6 %. Membrane NF-245 showed low ion rejection at pH 5, whereas the lactose at this
pH was nearly 99 %, that is why, this membrane is more able to separate lactose and monovalent
ions. Rejections of bivalent ions (70 % for NF-245 and 86 % for TFC-STR100) were close to the
lactose rejection which means, this method is not completely suitable for total whey desalination.

CONCLUSION
The average fluxes during natural whey ultrafiltration were 14 – 66 l.h-1.m-2. The influence of UF
pre-treatment on the velocity of following NF was minimal. The rejection of lactose on UF
membranes having cut off 100 and 500 nm was zero, and on the 5kDa membrane was 0.1 % only.
Three different types of flat polymeric membranes were compared (Desal DL (Osmonics), NF 245
(Dow) a TFC SR100 (Koch)). The average flux varied between 16 and 183 l/h.m2. Regarding the
kinetics, the most convenient membrane is NF-245 membrane which gives stable permeate flux
about 61 – 68 l.h-1.m-2 only slightly affected by the pH. The membrane TFC-SR100 gives the
highest permeate fluxes at pH 5.5 – 6, NF-245 at pH 5 – 6. Lactose rejection on all types
membranes was between 93-100%. Rejections of bivalent ions (70-86%) were close to the lactose
rejection which means, this method is not completely suitable for total whey desalination.

REFERENCES
[1] Suková I. 2006. In: Syrovátka v potravináĜství: Informaþní pĜehledy ÚZPI, Praha.
[2] Horton B.S. 1995. Commercial Utilization of Minor Milk Components in the Health and Food
Industries. Journal of Dairy Science, 78(11), 2584 – 2589.
[3] Groleau P.E., Lapointe J.F., Gauthier S.F. and Pouliot Y. 2004. In: International Dairy Federation
(Ed.). Advances in fractionating and separation: Process for novel dairy applications, Bulletin 389,
Brussels, Belgium, 85 – 91.

1696
 Separation and Fractionation of Aquilaria Malaccensis Oil Using Supercritical Fluid

Extraction and the Cytotoxic Properties of the Extracted Oil

A. H. Ibrahim1, S. S. Al-Rawi2, A. M. S. Abdul Majid1, N. N. Ab. Rahman3, K. M. Abo- Salah4, M. O.

Ab Kadir2

1
Department of Pharmacology, School of Pharmaceutical Sciences, Universiti Sains Malaysia, Penang,
Malaysia (aminmalikshah@gmail.com), (abraihimi@yahoo.com)
2
Department of Environmental Technology, School of Industrial Technology, Universiti Sains Malaysia,
Penang, Malaysia (akmomar@usm.my), (swsanrawi@yahoo.com)
3
Department of Biology, School of Distance Education, Universiti Sains Malaysia, Penang, Malaysia
(norulain@usm.my)
4
king Abdulla Institute for Nanotechnology, King Saud University, Riyadh, Saudi Arabia
(abusalah@ksu.edu.sa)

INTRODUCTION
Latest studies have started to be concerned in natural products from higher plants to isolate
and detect new active medicinal compounds. The importance role of these compounds have
expanded their widely applications to prevent and treat variety of human diseases. Aquilaria
species is aromatic plants commonly known as Gaharu wood in South East Asia [1]. The
essential oil of A. malaccensis is safe, simple and commonly used in traditional medicine to
relive pain, fever, rheumatism and asthma. Dash [2] extracted many compound from A.
malaccensis such as alkaloids, tannins, phenols, terpenoids, Quinones and Àavoenoids. Sice
there is no available study that investigates the cytotoxicity for A. malaccensis oil. Thus, the
aims of this study is to extract and fraction Aquilaria Malaccensis oil using supercritical fluid
extraction as a clean extraction method, and to investigate the cytotoxic properties of both the
extracted and fractioned oil.

MATERIALS & METHODS

Supercritical carbon dioxide was used to extract the oil of A. Malaccensis at temperature of 40-
50°C and set of pressure ranged 20.7, 27.6 and 34.5MPa, extraction dynamic time was 30 min.
The highest extracted sample was then fractionated using the best operating condition for
extracting the highest oil yield. Both the samples and the fractioned were tested for it cytotoxic
properties by employing MTT assay as an invitro study on the colon cancer cell HCT-116.

RESULTS & DISCUSSION

The result of this study showed that extraction yield increased with increasing both pressure
and temperature, and the highest extraction yield was 3.66g oil / 100g sample, at 50 ºC,
pressure 34.5 MPa, CO2 flow rate ” 1ml/ min, and particle size ” 500μm, dynamic time 30
minutes. [3] Reported that increasing temperature and pressure in supercritical can increased
the yield significantly. Moreover, increasing temperature increased extraction yield more than
increasing pressure while leaving the other parameters constant. Cytotoxic property of the
extracted oil of A. Malaccensis using supercritical was investigated by employing MTT assay

11th International Congress on Engineering and Food - Athens, Greece, 2011 1697
as an in vitro study on the colon cancer cell HCT-116. Result of this experiment showed that
the extracted sample at 50 ºC, pressure 20.7MPa, inhibited 99% of the colon cancer cell growth
at the concentration of 25μg/ml. However, temperature role was more crucial from pressure in
extracting the sample with the most cytotoxic. Under the best condition that give the highest
cytotoxic sample, the sample was fractioned into three fractions, and the three fractions were
tested for cytotoxicity using MTT assay. The result of this study showed that the first fraction
extracted at the first 10 min has the highest cytotoxicity property with 94% inhibition of colon
cancer cell growth. The IC50 for this fraction was carried out using six concentrations ranging
from 2.5, 5, 10, 15, 20 and 25μg/ml by employing the MTT assay as shown in figure 1.

Figure 1. The IC50 of A. Malaccensis oil extracted

using supercritical on the colon cancer cell HCT116.

The IC50 for this fraction was 3.5μg/ml which is representing the amount of drug that can kill
50% of the cell.

CONCLUSION
The results of this study revealed that the the supercritical extraction yield of A. Malaccensis
increased with increasing both pressure and temperature. Moreover, the temperature is more
crucial factor for the extraction yield as well as the property of the extracted sample as showed
from the result of this study increasing temperature increased the cytotoxic property of A.
Malaccensis oil on the colon cancer cell HCT116. Yet other investigations are now in process
to confirm its cytotoxicity and it use as an anticancer drug.

REFERENCES
[1] Takemoto, H., Ito, M., Shiraki, T., Yagura, T. and Honda, G. 2008. Sedative effects of vapor
inhalation of agarwood oil and spikenard extract and identification of their active components. J Nat
Med, 62, 41–46.
[2] Dash, M., Patra, J. K. and Panda P. P. 2008. Phytochemical and antimicrobial screening of extracts of
Aquilaria agallocha Roxb. African Journal of Biotechnology, 7 (20), 3531-3534.
[3] Hassan, M. N., Norulaini, N. A. N., Omar, A. K. M., & Ibrahim, M. H. 2000. Simple fractionation
through the supercritical carbon dioxide extraction of palm kernel oil. Journal of Separation and
Purification Technology, 19, 113–120.

1698
 Sugaring Out for Separation of Acetonitrile and Extraction of Proteins and Antibiotics

Pradip B. Dhamolea,b, Prafulla Mahajana, Hao Feng a,c

a
Energy Biosciences Institute, University of Illinois at Urbana-Champaign, Urbana, IL, USA
(haofeng@illinois.edu)
b
Department of Biotechnology, Sinhgad College of Engineering, Pune, India
c
Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign,
Urbana, IL, USA

INTRODUCTION
Liquid-liquid extraction, salting out, and reversed phase, high pressure liquid chromatography
(RP-HPLC) are typical methods for the separation of biomolecules, especially proteins and
antibiotics. Acetonitrile (ACN) is widely used as a solvent or mobile phase. It is miscible in all
proportions with water, which makes their separation a problem. Recent studies reported that
the addition of a monomeric sugar or a disaccharide to an ACN-water mixture created two
phases, one solvent-rich and the other aqueous, a phenomenon termed “sugaring out” [1-2]. In
this study, sugaring out was used for the separation of ACN from an aqueous phase and for the
extraction and recovery of selected biomolecules (BSA, trypsin, pepsin erythromycin,
streptomycin, and nalidixic acid). This method does not require a subzero temperature for the
phase partition, nor does it alter environmental conditions such as pH. Our earlier studies
investigated the phase separation at 1oC for relatively low sugar concentrations (15-50 g/L). In
the current work, the effects of different temperatures and glucose concentrations on sugaring
out were also studied.

MATERIALS & METHODS

Glucose at different concentrations was dissolved in DI water. The BSA, trypsin, pepsin,
streptomycin, and nalidixic acid were dissolved in the above sugar solutions while the
erythromycin (10 mg/mL) was dissolved in ACN before adding the sugar solutions. Sugar
solutions containing the hydrophilic biomolecules were mixed with ACN in a ratio of 1:1 (v/v).
The upper and lower phase volumes were recorded after phase separation. Samples were
collected using a disposable syringe and analyzed for solute (protein/antibiotic) concentration.
The results were expressed in terms of phase ratio (PR) and extraction (%).

RESULTS & DISCUSSION

Increasing the temperature from 6 to 18oC was accompanied by a gradual reduction in the
upper phase volume and hence a reduction in the phase ratio (Fig. 1a). When the temperature
was fixed, the upper phase volume and phase ratio increased with an increase in the glucose
concentration. Increasing the temperature lowered the ACN extraction efficiency (Fig. 1b). As
the sugar concentration was increased, the amount of ACN in the upper phase increased. The
maximum ACN recovery from the upper phase was 62%, which was achieved at the conditions
for the highest phase ratio. In the phase ratio vs. ACN plot (Fig. 1c), the ACN recovery
increases linearly with the phase ratio.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1699
BSA, trypsin, and pepsin were selected as model proteins in this study. All are hydrophilic in
nature. It was observed that more than 95% of the proteins were retained in the aqueous phase
(data not shown). An increase in the glucose concentration or temperature did not affect the
protein extraction. This shows that almost all of the proteins were retained in the aqueous
phase and very few (less than 2 %) were lost during the ACN separation process. Thus,
sugaring-out can be used for an effective recovery or purification of proteins from the ACN-
containing effluent of an RP-HPLC process.

The distributions of streptomycin, erythromycin and nalidixic acid were decided by their
hydrophobic/hydrophilic nature. For erythromycin, phase separation did not occur even with
120 g/L glucose at 18oC. 80-90% of the streptomycin and 91-94% of the nalidixic acid were
retained in the aqueous phase. On the other hand, 48-65% of the erythromycin was extracted
from the ACN-rich phase.

Figure 1. The effects of temperature and glucose concentration on sugaring out (1a); The effects of
temperature and glucose concentration on acetonitrile recovery (1b and 1c).

CONCLUSIONS
More than 95% (w/w) of the proteins were retained in the aqueous phase after the sugaring-out
separation of the ACN. Hydrophobic erythromycin (65% w/w) was extracted from the ACN–
rich phase and streptomycin (80-90% w/w) and nalidixic acid (91-94% w/w) were retained in
the aqueous phase.

REFERENCES
[1] Wang, B., Ezejias, T., Feng, H., Blaschek, H., 2008a. Sugaring-out: A novel phase separation and
extraction system. Chemical Engineering Science 63, 2595-2600.
[2] Wang, B., Feng, H., Ezeji, T., Blaschek, H., 2008b. Sugaring-out separation of acetonitrile from its
aqueous solution, Chemical Engineering Technology 31, 1869-1874.

1700
 Gamma-oryzanol Solubility and Effect of Solvents Mixture

Maitê S. Cuevasa, Regiane E. Shinzatoa, Mariana C. Costaa, Christianne E. C. Rodriguesb, Antonio J. A.

Meirellesa

a
University of Campinas, Campinas, Brazil (tomze@fea.unicamp.br)
b
University of São Paulo, Pirassununga, Brazil (chrisrodrigues@usp.br)

INTRODUCTION
Gamma-oryzanol is a complex mixture of triterpenic alcohols and phytosterols esterified with
ferulic acid. This compound has been reported as a powerful antioxidant and
hypocholesterolemic agent in several scientific studies [1,2]. This nutraceutical compound can
be extracted from rice bran oil through oil saponification followed by solvent extraction and
crystallization [3]. Despite the functional importance of this compound, there are no studies
reporting the gamma-oryzanol solubility in organic solvents that could allow the extraction or
preservation of this substance in the rice bran oil.
Hexane is commonly used as solvent for rice bran oil extraction and the knowledge of gamma-
oryzanol solubility in this solvent is an important step to preserve rich fractions of oil. In
addition, mixture of solvents can increase the gamma-oryzanol solubility as suggested by
Scatchard-Hildebrand theory [4].
The main aim of this study was to investigate the gamma-oryzanol solubility in pure solvents
(hexane and hexanol) as well as in two binary mixtures of these solvents with mass ratios of
1:1 and 3:1, from 283.2 to 323.2 K. The solid-liquid equilibrium data were correlated using the
modified Apelblat equation.

MATERIALS & METHODS

Solvent and solute in excess were added to glass jacketed cells. The temperature in each cell
was controlled by circulating thermostatic water in the jacket and was considered to be
accurate within ± 0.1 K. At a pre-set temperature the mixture was stirred for 30 min and left to
rest for at least 24 h. The composition of gamma-oryzanol in the liquid phase was determined
using gravimetric method. The sample was dried at 378.2 K until constant mass. After that, the
solid-liquid equilibrium data of gamma-oryzanol was correlated using the Apelblat equation
[5].

RESULTS & DISCUSSION

Figure 1 shows the experimental data, obtained in the temperature range from 283.2 to 323.2
K, and calculated data by modified Apelblat equation. It can be observed that the solubility of
gamma-oryzanol increases with the increasing of temperature.
It is also possible to note that the compound studied presented lower solubility in hexane than
in hexanol. In addition, it is observed that gamma-oryzanol has its solubility increased when
mixed solvents are used, especially in the 1:1 mass ratio of hexane: hexanol.
Scatchard Hildebrand theory predicts that the solubility of a solid has a maximum value in that
solvent whose solubility parameter is the same as that of the solute. In general, the equation

11th International Congress on Engineering and Food - Athens, Greece, 2011 1701
proposed by Apelblat can describe the temperature dependence of solubility of gamma-
oryzanol in the proposed solvents.

-2

-4
lnx2

-6 hexane
hexanol
hexane-hexanol 1:1
hexane-hexanol 3:1
Apelblat
-8
0.0030 0.0032 0.0034 0.0036
-1
1/T (K )

Figure 1. Experimental and calculated solubilities of gamma-oryzanol in different organic solvents.

CONCLUSION
Solubility data of gamma-oryzanol in pure solvents (hexane and hexanol) and in two binary
mixtures with mass ratios of 1:1 and 3:1 in the temperature range from 283.2 to 323.2 K were
determined. It was observed that the solubility of nutraceutical compound increase with rising
temperature and in the alcohol solvent. It was also observed that the mixture of the two pure
solvents presented a synergetic effect improving the solubility of the compound, being that this
behavior can be explained by the Scatchard Hildebrand theory. In general, the modified
Apelblat model can describe the temperature dependence of solubility of gamma-oryzanol.

ACKNOWLEDGEMENTS; The authors wish to acknowledge FAPESP (09/17855-3,

07/06170-4, 08/56258-8), and CNPq (140642/2010-2) for the financial support.

REFERENCES
[1] Seetharamaiah G.S. & Chandrasekhara N. 1993. Comparative hypocholesterolemic activies of
oryzanol, curcumin and ferulic acids in rats. Journal of Food Science and Technology, 30 (4), 249-
252.
[2] Deckere E.A.M. & Korver O. 1996. Minor constituents of Rice bran oil as functional foods. Nutrition
Reviews Nutrition Reviews, 54 (11), 120S-126S.
[3] Rao, K.V.S.A., Rao, B.V.S.K. & Thengumpillil, N.B.K. 2002. Process for the Isolation of Oryzanols
from Rice Bran Oil Soap Stock. U.S. Patent 6410762.
[4] Prausnitz, J.M., Linchtenthaler, R.N. & Azevedo E.G. 1999. Molecular Thermodynamics of Fluid-
Phase Equilibria. Prentice Hall, New Jersey, USA.
[5] Liu C.W. & Fu A.W. 2004. Solubility of Niacin in 3-Picolin + Water from 287.65 K to 359.15 K.
Journal of Chemical & Engineering Data, 49(1), 155-156.

1702
Extraction of ascorbic acid using alcohol/phosphate potassium salt – based aqueous two-

phase system

Igor A. O. Reisa, Samuel B. dos Santosa, Ludimila A. S. Nascimentoa, Naiana Oliveiraa, Sónia P.M.
Venturab, João A.P. Coutinhob, Cleide M.F. Soaresa,c, Álvaro Silva Limaa,c,*

a
Universidade Tiradentes, Aracaju-Sergipe, Brasil
b
Universidade de Aveiro, Aveiro, Portugal
b
Instituto de Tecnologia e Pesquisa, Aracaju-Sergipe, Brasil, *E-mail: alvaro_lima@unit.br

INTRODUCTION
The extraction and partial purification of bioactive molecules is a field of great interest to the
pharmaceutical and bioengineering. To achieve these purposes, some solvents with different
polarities such as ethanol, methanol, chloroform and acetone are used. However, the yield and
purity of the interest products is relatively low, therefore the solvent consumption is inevitably
high, and therefore the process costs are also high. An effective and economically viable
method for the separation and purification of biomolecules is the partitioning in aqueous two-
phase system (ATPS). The system consists of two immiscible liquid phases above a critical
concentration. The technique is widely used because of their low-coast, biocompatibility and
easy scale up, allowing a selective and rapid extraction process. Ascorbic acid is a powerful
antioxidant found in citrus fruit, which presents as white crystalline solid, odourless and water
soluble. The goal of this work is to determine the application of aqueous two-phase system
(alcohol/phosphate potassium salt) as a step to extract ascorbic acid.

MATERIALS & METHODS

The binodal data was determined for the studied system through the cloud point titration
method at 298.15 using alcohols and phosphate salts. The ternary system compositions were
determined by the weight quantification of all components added. The tie-lines and the tie-line
lengths were also represented for this system. The tie-lines (TLs) were determined by a
gravimetric method adopted from Merchuck et al. [1]. Each individual TL was determined by
application of the lever arm rule. The concentration of ascorbic acid was measured using the
Tillmans method. The partition coefficient of L-ascorbic acid K, is defined here as the ratio of
the concentration of ascorbic acid in the alcohol and salt-rich phases.

RESULTS & DISCUSSION

The binodal curves indicate that the alcohol with larger linear alkyl chain, the greater is the
ability for ATPS formation. It is well-know that the solubility of an aliphatic alcohol in water
and the mutual miscibility depend on the chain length, and decrease with increasing number of
carbon atoms in the chain [2]. The relation between the linear alkyl chain and alcohol’s
hydrophobicity nature is directly proportional. The branched-chain alkyl alcohol had a
negatively effect on the formation of the ATPS, due to higher polarity of 2-propanol as
compared with that of 1-propanol. The capability for creating the ATPS followed the
Hofmeister series: K3PO4 > K2HPO4 > K2HPO4/KH2PO4 [3]. The aqueous solutions of

11th International Congress on Engineering and Food - Athens, Greece, 2011 1703
potassium phosphate salts confer different pH values for each system, which are 13.7; 9.1 e 7.0
for K3PO4, K2HPO4 and K2HPO4/KH2PO4, respectively. The binodals curves were fitted using
the approach described by Merchuck et al. [1]. Almost all systems have R2 values above 0.99,
indicating good model fit to experimental data. For all systems the top-rich phase is the
alcohol-rich phase and the bottom phase is the potassium phosphate salts-rich phase.
The L-ascorbic acid was preferentially partitioned to bottom phase (K<<1.0). L-ascorbic acid
solubility increases in the solvent of higher polarity in the order of 2-propanol, ethanol,
methanol and water [4]. On the other hand, the increase of linear alkyl chain length and
consequently the hydrophobicity of alcohol, for systems with K2HPO4 and buffer solution,
increase the K. However, by Wang et al. [5], the interaction ion-dipole (in microcosms)
between salt and water molecules leads to the hydratation of ions, and then the phase-forming
slats can dissolve in the bottom phase; meanwhile, the above interactions decrease the amount
of free water molecules in the bottom phase, and it leads to the exclusion of alcohol and target
to top phase. The alkyl chain ramification of alcohol does not modify the K of L-ascorbic acid.
The decrease in pH value of system, due to potassium phosphate salt, increases the partition
coefficient of L-ascorbic acid for all cases. Depending on the pH value of the system, L-
ascorbic acid will exist in two forms: when the pH of system is acid, the non dissociated
(neutral charge) form will be dominant, on the other hand at alkaline pH the dissociated
(negative charge). Probably the dissociated form trend to migrate to bottom phase decreasing
the partition coefficient. But in non dissociated form, the phase more hydrophobic (top phase)
can attract the nucleus of five atoms of L-ascorbic and the K increase. The best value for K was
achieved by the buffer solution (neutral pH), in which the L-ascorbic acid is more stable.

CONCLUSION
The systems with aliphatic chain promote the best phase separation, and the K2HPO4/KH2PO4
buffer has a lower ability for the ATPS formation. The TL were satisfactorily correlated with
Merchuck’s equations. The L-ascorbic acid was partitioning to bottom phase, but decreasing of
pH value of system, a little portion of L-ascorbic acid trend to migrate to top phase.

REFERENCES
[1] Merchuk, J.C., Andrews, B.A. & Asenjo, J.A. 1998. Aqueous two-phase systems for protein
separation: Studies on phase inversion. Journal of Chromatography B: Biomedical Sciences and
Applications. 711, 285-293.
[2] Ooi, C.W., Tey, B.T., Hii, S.L., Kamal, S.M.M., Lan, J.C.W., Ariff, A. & Ling, T.C. 2009.
Purification of lipase derived from Burkholderia pseudomallei with alcohol/salt-based aqueous two-
phase systems. Process Biochemistry, 44, 1083–1087.
[3] Golubitskii, G.B., Budko, E.V., Basova, E.M., Kostarnoi, A.V. & Ivanov, V.M. 2007. Stability of
ascorbic acid in aqueous and aqueous–organic solutions for quantitative determination. Journal of
Analytical Chemistry, 62, 742–747.
[4] Shalmashi, A. & Eliassi, A. 2008. Solubility of L-(+)-ascorbic acid in water, ethanol, methanol,
propan-2-ol, acetone, acetonitrile, ethyl acetate, and tetrahydrofuran from (293 to 323) K. Journal of
Chemical Engineering Data, 53, 1332-1334.
[5] Wang, Y., Han, J., Xu, X., Hu, S. & Yan, Y. 2010. Partition behavior and partition mechanism of
antibiotics in ethanol/2-propanol–ammonium sulfate aqueous two-phase systems, Separation and
Purification Technology, 75, 352–357.

1704
 Partition of Amyloglucosidase in poly(ethyleneglycol) / sodium-poly(acrylate) Aqueous
Two-Phase Systems

L.A.P. Alcântaraa, L.A. Minima*, C.A. Mourãoa, V.P.R. Minima

a
Federal University of Viçosa, Viçosa,Brazil (*E-mail: lminim@ufv.br)

INTRODUCTION
Amyloglucosidade (exo-1,4-Į-glucosidadese, E.C. 3.2.1.3) is an amylolitic enzyme of high
commercial value widely used in the food industry as well as in the starch processing, baking,
brewing and distilling industry [1]. Reducing cost and enzyme losses in the purification
process are of fundamental importance in the viability of the process [2]. Liquid-liquid
extraction by aqueous two-phase systems (ATPS) has proved to be an important tool for
separating and purifying mixtures of biomolecules by extraction. The advantages of aqueous
two-phase extraction compared to other purification methods lie in high selectivity, low cost
and can be suitable for continuous operation in large scale, thus allowing wider
biotechnological applications [2]. These systems are suitable for purification of biological
material as the phases contain 70% to 90% water, thus reducing the denaturation of
biomolecules [3]. Therefore, the aim of this work was to evaluate the influence of pH and
concentration of polyethylene glycol (PEG) and sodium polyacrylate (NaPA) on the partition
of amyloglucosidase (AMG). In order of optimizing the partition of this enzyme in PEG/NaPA
aqueous two-phase systems a response surface technique was employed.

MATERIALS & METHODS

The experiments were performed using poly (ethylene glycol) (4000 g·mol-1) and sodium
polyacrylate (15000 g·mol-1) at different conditions of pH (6.0-7.0), PEG concentration (11.0%
-12.0% w/w) and NaPA concentration (10.4% - 11.4% w/w), at 298.15 K. The concentration of
sodium chloride (NaCl) has been fixed at 0.2 mol·L-1. The experiments were conducted
employing a face-centered design (FCD), added of 4 central points. The partitioning
experiments were prepared in 10 mL centrifuge tubes using 2.0 mL of upper and lower phases
of a pre-equilibrated ATPS. A volume of 100 ȝL of enzyme solution (269.18 units·mL-1 of
amyloglucosidase) were added in the tubes, which were thoroughly mixed, centrifuged at
4000 × g for 15 min and incubated in a water bath of constant temperature for 6 hs. The
enzymatic activity of amyloglucosidase in the top and bottom phases were determined
according to Tanuja et al. [4]. Reducing sugars released during the enzymatic reaction were
quantified by the method dinitrosalicylic acid [5]. The amyloglucosidase partition coefficient
(KAMG) is defined as the ratio of the volumetric activity in the top phase (At) to that in the
bottom phase (Ab), according to the Eq.1.
At
K AMG (1)
Ab
RESULTS & DISCUSSION
A FCD design was employed in order to evaluate the effect of pH and NaPA concentration
(CNaPA) on the partition behavior of the AMG. A detailed representation of the KAMG from the
experimental results is presented as contour plot in Figure 1.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1705
Figure 1. Effect of pH and NaPA concentration on the partition coefficient of amyloglucosidade.

The experimental results were submitted to analysis of variance and showed that partition of
AMG was most suitably described using a quadratic polynomial model. KAMG was influenced
by the linear and quadratic contribution of pH and NaPA (p<0.05). Only the contribution of
PEG was not statistically significant (p>0.05). The model of AMG partition is described in the
Eq.2.
y  15 . 303  35 . 673 x 1  18 . 506 x 2  2 . 715 x 12  0 . 855 x 22 (2)
where y is the AMG partition coefficient (KAMG), x1 and x2 is the values for pH and
concentration of NaPA (CNaPA), respectively. The value o the determination coefficient (R2 =
0.916) confirmed the suitability of the model.

CONCLUSION
The feasibility of using ATPS constituted of PEG (molar mass of 4000 g·mol-1) and NaPA
(molar mass of 15000 g·mol-1) for the AMG purification was shown in this work. The
partition coefficient values was found in the range of 1.8 to 2.0 and it was verified that the
AMG partition are mainly affected by pH and the concentration of NaPA.

REFERENCES
[1] Guzman-Maldonado, H., & Paredes-Lopez, O. 1995. Amylolytic enzymes and products derived from
starch: a review. Critical Reviews in Food Science and Nutrition, 35, 373–403.
[2] Diamond, A.D., & Hsu, J.T. 1992. Aqueous two-phase systems for biomolecule separation. Advances
in Biochemical Engineering Biotechnology, 47, 89–135.
[3] Albertsson, P.A. 1986. Partition of Cellular Particles and Macromolecules (3rd ed.). Wiley
Interscience, New York.
[4] Tanuja, S., Scrinivas, N.D., Rao, K.S.M.S.R., & Gowthman, M.K. 1997. Aqueous two phase
extraction for downstream processing of amyloglucosidase. Process Biochemistry, 32(8), 635-641.
[5] Miller, G.L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugars. Analytic
Chemistry, 31 426-428.

1706
11th International Congress on Engineering and Food - Athens, Greece, 2011 1707
subsequently temperature maintained on 35.5 °C, 44.8 °C, 54.5 °C and 62.5 °C. The feed
mixture has been injected for 3 min. The flow rate was 1.5 l.h-1. The maximum mannose purity
77.5 % was achieved 62.5 °C. On the base of graphs obtained there were evaluated parameters
which are necessary to evaluate the HETP (height equivalent to theoretical plate). In examined
range of temperature the HETP was lowest at 62.5 °C, so here we can in find the optimum
separation conditions for glucose-mannose separation.

Continuous separation
The optimal temperature 62.5 °C for SMB separation was chosen according discontinuous
measurements. The operational parameters were obtained using special software [5] from
chromatographic curves of discontinuous measurements at optimal temperature. The values of
operational parameters were verified with simulation program [5]. The SMB system was
started with predicted parameters and there were collected fractions in extract and raffinate
during the steady state. Purity in each stream was calculated according the content of both
substances in streams. The purity of mannose achieved in experiment 71.2 %, in simulation
88.6 %. The setting and maintaining of continuous mode demand very precious flow meters in
this case of near retention times of both separated compound. There could be the drop of
maximum purity between discontinuous and continuous separation.

CONCLUSION
The mixture of mannose and glucose was separated using gel-type catex resin in Na form
(Lewatit MDS 1368 Na). The discontinuous measurements on two serially connected columns
were performed at four different temperatures 35.5 °C, 44.8 °C, 54.5 °C and 62.5 °C. The
maximum purity 77.5 % and also lowest HETP were achieved at 62.5 °C in the case of
discontinuous separation. The maximum purity achieved in the continuous measurement was
71.2 %.

REFERENCES
[1] Koeckritz A., Kant M., Walter M. & Martin A. 2008. Rearrangement of glucose to mannose catalysed
by polymer-supported Mo catalysts in the liquid phase. Applied Catalysis A: General, 334(1-2), 112-
118.
[2] Thomas J. & Lobel L.H. 1976. Chromatographic separation of glucose and mannose on cation-
exchange resin. Analytical Biochemistry, 73(1), 222-226.
[3] Sherman J.D. & Chao C.C. 1984. Separation of mannose by selective adsorption on zeolitic molecular
sieves. US Patent 4 471 114.
[4] Kulprathipanja S. 1989. Process for separating glucose and mannose with CA/NH4-exchanged ion
exchange resins. US Patent 4 837 315.
[5] Henke S., Bubník Z. & Kubát M. 2006. Modelling, simulation and design of SMB apparatus. 2nd
Technical Symposium of CIGR Section VI future of Food Engineering, Warszawa, Poland, April 26-
28, 2006. Proceedings p.223.

1708
 Brewer’s spent grain standardization and upstream processes for enzymatic hydrolysate

production

Catalina E. Kotlar a, b, Mariela Belagardi a, c, María V. Agüero a, b and Sara I. Roura a, b

a
Grupo de Investigación en Ingeniería en Alimentos, Departamento de Ingeniería Química y en Alimentos, Facultad
de Ingeniería, Universidad Nacional de Mar del Plata, Mar del Plata, Argentina
b
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina
c
Licenciatura en Nutrición, Universidad F.A.S.T.A., Mar del Plata, Argentina

INTRODUCTION
A wide range of agro-industrial by-products are available in large quantities which have considerable
nutritional potential. Brewery waste is a typical example of such unrealized potential. Value-added
products are increasingly being sought from brewer’s spent grain (BSG), a protein rich residue generally
used as livestock diets. One hundred kilograms of barley produces 170 kg of wet BSG (having 80–85%
moisture). To determine the potential use of a substrate is necessary to realize its correct standardization.
The lack of protein solubility is one of the limitations for their extensive use in food processing. In order
to use the residue BSG as a substrate for growth and enzyme production of proteolytic bacteria, the main
objectives were to standardize the BSG used as raw material and to define the fermentation upstream
processes for enzymatic hydrolysis by Bacillus cereus. This work is of usefulness to increase the added
value by generating protein hydrolyzates with potential use in various industries.

MATERIALS & METHODS

The composition characterization, granulometry assay, size reduction analysis and the microbiological
stability of different lots and varieties of BSG provided by a local company (Antares S.A., Argentina)
were carried out. Simple pre-treatments of the substrate: granulometry and grinding assays, polyphenols
extraction (after alkali hydrolysis by Folin-Ciocalteu reagent and using gallic acid as standard) and
sterilization were defined in order to eliminate interference and standardize the substrate accordingly.
After that the BSG was hydrolyzated in submerged fermentation with Bacillus cereus. Antagonistic and
synergistic effects were evaluated by combinations of strains as inoculum.

RESULTS & DISCUSSION

The total bacteria count of dry BSG remained almost constant during the overall storage period. Wet BSG
microorganism load achieved a final CFU value five orders higher than BSG previously dried. The initial
microflora was predominantly termophilic aerobic bacteria. Drying was the better alternative for BSG
preservation. BSG showed small variations in composition. The water, lipids and ash contents differed
significatively with the BSG varieties (Table 1). The raw material was dried at 60ºC for 24-48 h and
sieved. BSG retained above mesh Nº10 was ground, then polyphenols were extracted with an alcohol:
water solution 30:70 for 60 min in an orbital shaker at 50 rpm and finally BSG was added to Mineral salt
Medium (MSM) and sterilized in autoclave for 15 min at 121ºC. The obtained material was termed
fermentation substrate (FS).
Different fractions of FS (w/v) were fermented through B. cereus activity and the evolution of soluble
protein content (SPC) during the fermentation period was analyzed. Firstly an increase in the SPC was
observed as the FS concentration increases during the overall incubation period (30 h), achieving at 36%
w/v of FS the higher SPC. At higher FS concentration, the SPC decreased. Higher FS concentration

11th International Congress on Engineering and Food - Athens, Greece, 2011 1709
could increase the viscosity of the medium which possible resulted in oxygen limitation for bacterial
growth, restricting the production of protease by B. cereus and then decreased the soluble protein content.
Fermentation of FS was carried out using microbial culture or crude enzymatic extract and SPC was
measured during the incubation time. The higher SPC was achieved by inoculating 5% v/v of crude
enzyme; approximately a 20% lower SPC was found when the same concentration of fresh culture was
used, supporting the advantage of using enzymatic extract vs. Microbial culture.

Table 1. Proximate composition of BSG varieties

BSG variety
Assays
100 % Pilsner 93% Pilsner 78% Pilsner
Moisture (%w/) b b
77,40 ± 0.28 77,69 ± 0.30 79,70 ± 1.18a
Total Ash (%w/w) DB1 a a
2,58 ± 0.128 2,21 ± 0.49 2,11 ± 0.38a
Crude Protein (%w/w) DB a a
29,97 ± 0.93 30,52 ± 0.43 34,55 ± 0.49a
Fat (%w/w) DB
5,82 ± 0.93b 5,67 ± 0.79b 6,74 ± 2.74a
Carbohydrates (%w/w) DB
61,55 ± 1.12a 60,97 ± 0.79a 56,29 ± 2.748b
1
DW: dry weight
The number marked with different superscript letters in the same row meant statistic difference at significant level 5%.

Looking for synergistic effects of B. cereus with other proteolytic microorganisms, a notorious synergistic
effect was observed when B. cereus was inoculated either with Pseudomonas pútida or Pseudomonas not
classified (Table 1).
Additionally, the proteolytic activity displayed a similar trend to protein solubility, confirming the
presumption of Chen et al. that protease was induced to breakdown the BSG into soluble protein to
support the microorganism growth.

Table 1. Proteolytic activity measured at 24 h from FS fermentation medium (36 %w/v) with mixed inoculum at 32ºC
in an orbital shaker at 60 rpm.
Inoculum Proteolytic activity (Abs 420 nm)
2.5% v/v B. cereus + 2.5% v/v Pseudomonas not classified 0,254 ± 0.027a
2.5% v/v B. cereus + 2.5% v/v Pseudomonas pútida 0,264 ± 0.011a
2.5% v/v B. cereus + 2.5% v/v Enterococcus hirae (a) 0,128 ± 0.005 c
2.5% v/v B. cereus + 2.5% v/v Enterococcus hirae(b) 0,125 ± 0.006 c
2.5% v/v B cereus + 2.5% v/v Lactococcus lactis subsp. lactis 0,098 ± 0.009 d
5% v/v Pseudomonas not classified 0,102 ± 0.005 d
5% v/v Pseudomonas pútida 0,095 ± 0.002 d
5% v/v Enterococcus hirae (a) 0,206 ± 0.012 b
5% v/v Enterococcus hirae (b) 0,134 ± 0.006 c
5% v/v Lactococcus lactis subsp. lactis 0,121 ± 0.008 c
5% v/v B. cereus 0,131 ± 0.009 c
The number marked with different superscript letters in the same row meant statistic difference at significant level 5%.

CONCLUSION
This study provides a fundamental work for the production of hydrolyzates and the conversion of BSG to
soluble protein through submerged fermentation process by Bacillus cereus. This methodology could be
applied on a large scale and extended to other agro industrial wastes.

1710
 Treatment of passion fruit juice by membrane process technology

Rui Dominguesa, Grasiele Madronab Vicelma Luiz Cardosoa, Miria H. M. Reisa

a
Federal University Of Uberlandia, Chemical Engeneering Faculty, Uberlândia-MG, Brazil (miria@feq.ufu.br)
b
State University of Maringa, Food Engeneering School, Maringá-PR, Brazil (grasiele@yahoo.com.br)

INTRODUCTION
Traditional stabilization methods, such as thermal pasteurization, are generally applied for fruit juice
processing. However, heat processing for yellow passion fruit results in change of its aroma and flavor.
Membrane processes have been studied for clarification and concentration of fruit juices. However,
fouling is the major constraint during separation using membranes. Its occurrence leads to a decline in
membrane permeability. Matta et al. indicated that the use of enzymatic pretreatment in fruit juice may
increase membrane fluxes, since the presence of cell-wall polysaccharide compounds is the main cause of
fouling occurrences in juice filtration. Vaillant et al. studied the crossflow microfiltration of passion fruit
juice using ceramic membranes with 0,2 μm average pore size. The scope of the present work was to
evaluate different conditions of enzyme pretreatments for yellow passion fruit juice, aiming to minimize
the juice viscosity for further membrane filtrations. After that, membrane filtrations were carried out in
order to observe the applicability of this process for juice clarification.

MATERIALS & METHODS

The pulp of passion fruit was purchased from a local pulp industry (Minas Gerais – Brazil). It was stored
at -16ºC and defrosted to room temperature before use. Enzymatic treatments proposed in this word
applied different available commercial enzymes with different activities. Bacterial Amylase, Celluclast,
and Pectinex 3X L were purchased from Novozymes, and Pectinase from Aspergillus niger was
purchased from Sigma-Aldrich. Two enzymatic mixtures were also evaluated, by mixing equal quantities
of different enzymes: Amylase + Celluclast + Pectinex (mixture 1) and Pectinase + Amilase (mixture 2).
In order to investigate the action of each one of these enzymes, preliminary tests were conducted with
these enzymes (pure and mixed) for 60 minutes in a 250 mL conic flasks at constant agitation and
temperature (200 rpm and 50ºC, respectively), using a concentration of 5 mL/L of each enzymatic
preparations. Using the most promising enzyme observed in preliminary tests, a factorial planning was
carried out in order to observe which variables most influence the process. These analyses were done
using the software STATISTICA 7.0. The microfiltration unit used in this work was purchased from
PAM (Rio de Janeiro, Brazil). It works with polyetherimide (PEI) hollow fibber membranes, with total
effective filtration area of 0,056 m2 and 0,4 μm average pore diameter. Microfiltrations were carried out
with 3L batches of passion fruit at room temperature and at 1, 2, and 3 bar. The fruit pulp was previously
centrifuged at 12000 rpm during 4 minutes. After that, the centrifuged juice was treated at the best
condition of the enzymatic treatment.Color analyses were carried out by collecting the absorbance at 540
nm using a Shimadzu UV 1240 spectrophotometer. Soluble solids were measured with a calibrated hand
refratometer and turbidity was measured with a Nova Organica HD 144 turbidimeter. Viscosities were
measured by using a Brookfield LVDV-III digital rheometer at 25º at a 303 s-1 shear rate.

RESULTS & DISCUSSION

Enzymatic Treatment
The obtained results with the different proposed enzymes for passion fruit juice treatment showed that the
enzymatic preparations of Pectinex 3X L, Mixture 1, and Mixture 2 resulted in higher viscosity

11th International Congress on Engineering and Food - Athens, Greece, 2011 1711
reductions. Taking availability and economic reasons into account, Pectinex 3X L, which contains
pectinolityc, cellulase, and amylase activities, was chosen for use in subsequent experiments in this work.
The effect of enzyme concentration was evaluated by treating the samples with different
concentrations of Pectinex 3X L at 50ºC for 60 min. The results have shown that viscosity
removal is function of the concentration. However, runs with enzyme concentrations higher
than 1 mL/L did not show increase in viscosity reduction. Similar results were observed by
Vaillant et al.. The effect of temperature was evaluated by testing enzymatic reactions on
several temperatures, using an enzymatic concentration of 1 mL/L at 60 min of incubation
time. The obtained results showed that 50°C is the best temperature for enzymatic treatment of
passion fruit juice. The obtained results with the factorial planning 23 showed that an increase on
viscosity reduction is observed with the temperature increasing. The enzyme concentration is also a
significant variable, as the viscosity is also reduced with enzyme concentration increasing. Temperature
and enzyme concentration have positive effects on viscosity reduction, and are significant at p<0.05. The
standardized effect estimates show that the temperature has a higher effect in comparison to the enzyme
concentration. The time of incubation did not seem to be significant in the process, and neither any of the
interactions between the observed variables at p<0.05. Practical questions indicate the application of a
minimum of 60 min incubation time in order to guarantee the complete reaction, as suggested by Vaillant
et al..
Microfiltration experiments
Results presented in Figure 1 show that the permeate flux reduced after 20 min of filtration. This behavior
was also observed by Yu et al. for passion fruit juice ultrafiltrations. It can be also observed that the
higher pressures slightly increases the permeate flux.
21 Table 1. Parameter reductions after the microfiltration
19 process in the clarified juice in comparison with the
17
enzymatic treated juice
F lu x (L h -1 m -2 )

15 P= 2 bar
Soluble
13 P= 3 bar Pressure Viscosity Color
11 P= 1 bar Turbidity (%) Solids
(bar) (%) (%)
9 (%)
7
1 99.8 16.9 97.8 17.9
5
0 20 40 60 80 100 120 140 160 180 200 220 2 99.7 22.7 98.4 4.2
Time (min)
3 99.1 27.9 97.3 16.0
Figure 1. Permeate fluxes after microfiltration
mean 99.5 22.5 97.8 12.7
processes at 1, 2, and 3 bar.

The results presented in Table 1 show that the microfiltration process reduces significantly the juice’s
color and turbidity. The applied pressure does not influence significantly these removals, although it
seems to have a trend that color and turbidity reductions are higher in smaller pressures. Membrane
process does not reduce significantly the viscosity and the soluble solute.

CONCLUSION
Pectinex 3X L has proved to be the most efficient enzyme for viscosity reduction of passion fruit juice,
among the enzymes evaluated in this work. The 23 factorial planning showed that reaction time did not
influence viscosity reduction of passion fruit juice within the analyzed time intervals of 30 to 120 min.
Filtration experiments showed that the microfiltration is a suitable process for passion fruit juice
clarification.

1712
 Honey & Honey Adulteration Detection: A Review

Laleh Mehryara, Mohsen Esmaiilib

a
Department of Food Science and Technology, University of Urmia, Iran (laleh.mehryar@gmail.com)
b
Department of Food Science and Technology, University of Urmia, Iran (m.esmaiili@urmia.ac.ir)

INTRODUCTION
Honey is an ancient valuable food and in most cases has enchanted its consumers by its medic
characteristics. It consists mainly of sugars. Honey composition according to the studied
literature is mainly dependant on its floral source and differs in various honeys. The dietary
frauds in particular the adulteration are practices in constant progress. Adulteration consists of
adding external chemical substance(s) into a food product that contains naturally similar
substance(s). Honey adulteration appeared on the world market in the 1970s when high-
fructose corn syrup was introduced by the industry. Many foods have the potential to be
deliberately adulterated, but those that are expensive and are produced under wide fluctuations
in weather and harvesting conditions are particularly susceptible; honey is one of such
material. Although the adulteration of honey is not injurious to health, problems of honey fraud
negatively influence market growth by damaging consumer confidence. It seems quite
necessary that preparing an overall review of the applied procedures by researchers in detecting
honey adulteration would be useful and serve as a good source in oncoming works.

HONEY ADULTERATION
Analysis methods
Gas Chromatography (GC) and Liquid Chromatography (LC) analysis: This method may
be considered as a replacement of isotopic analysis, which has some limitations.

Near Infrared Transflectance Spectroscopy (NIR): It is a rapid, non-destructive and

relatively inexpensive method which may be suitable for use as a screening technique in the
quality control of honey [1].

Fourier Transform Infrared (FTIR) spectroscopy with Attenuated Total Reflectance

(ATR): In contrast to the time-consuming carbon isotope ratio analysis techniques, these FTIR
spectroscopic procedures can be performed in very short time [2].

Protein characterization: The major proteins in honey have different molecular weights
depending upon the honeybee species. Therefore, the measurement of major proteins in honey
is a useful method to discriminate the honey that produced from different honeybee species.

High-Performance Anion-Exchange Chromatography with Pulsed Amperometric

Detection (HPAEC-PAD): It is an efficient tool for the characterization of the honey floral
species. This method is less time consuming and less expensive than other methods [3].

11th International Congress on Engineering and Food - Athens, Greece, 2011 1713
Liquid Chromatography Coupled to Isotope Ratio Mass Spectrometry (HPLC-IRMS):
The new procedure has advantages over existing methods in terms of analysis time, sensitivity,
lack of sample preparation, reduced consumption of reagents, and simplicity of the operative
procedure. In addition, it is the first isotopic method developed that allows beet sugar addition
detection [4].

Calorimetric methods (Application of DSC): Application of DSC showed the possibility of

using the glass transition temperature to distinguish between honeys and syrups and is a
powerful technique for characterizing the thermal behavior of honeys and for detecting the
effect of adulteration on physicochemical and structural properties of samples.

Stable Carbon Isotope Ratio Analysis (SCIRA): It is determined by the 13C/12C isotope ratio,
which is different in C4 or CAM plants, when compared to C3 plants [2].

Fourier Transform (FT) Raman spectroscopy: FT-Raman spectroscopy is successfully

applicable to detect beet and cane invert syrups. This method can also be used to discriminate
between the types of adulterants irrespective of its floral origin [5].

Microscopic detection: Microscopic analysis of adulterated honeys with cane sugar exhibited
parenchyma cells, single ring vessels and epidermal cells. Overall the microscopic procedure is
a good screening method for the detection of adulteration of honey with cane sugar products.

CONCLUSION
To sum up, according to the obtained data from studied literature it is to some extent obvious
that nearly the majority of physicochemical characteristics of honey depend on floral source.
Honey adulteration is a critical problem which is determined by various techniques to get
information from each aspect of it. Based on the type of adulterants each applied method seems
to be beneficiary by itself.

REFERENCES
[1] Kelly J.D., Petisco C. & Downey G. 2006. Potential of near infrared transflectance spectroscopy to
detect adulteration of Irish honey by beet invert syrup and high fructose corn syrup. J. Near Infrared
Spectrosc. 14, 139-146.
[2] Gallardo-Velázquez T., Osorio-Revilla G., Zuñiga-de Loa M. & Rivera-Espinoza Y. 2009.
Application of FTIR-HATR spectroscopy and multivariate analysis to the quantification of
adulterants in Mexican honeys. Food Research International 42, 313–318.
[3] Morales V., Corzo N. & Sanz M.L. 2008. HPAEC-PAD oligosaccharide analysis to detect
adulterations of honey with sugar syrups. Food Chemistry 107, 922–928.
[4] Cabañero A.I., Recio J.L. & Rupérez M. 2006. Liquid Chromatography Coupled to Isotope Ratio
Mass Spectrometry: A New Perspective on Honey Adulteration Detection. J. Agric. Food Chem. 54,
9719-9727.
[5] Paradkar M.M. & Irudayaraj J. 2001. Discrimination and classification of beet and cane inverts in
honey by FT-Raman spectroscopy. Food Chemistry 76, 231–239.

1714
 Scaling-up effects on supercritical CO2 extraction kinetics of pelletized tomato

Gonzalo A. Núñeza, Lorena I. Mödingera, José M. del Vallea,*, & Rudolf Eggersb

a
Dept. Chemical & Bioprocesses Engineering, Pontificia Universidad Católica de Chile, Santiago, Chile
(*email: delvalle@ing.puc.cl)
b
Inst. Thermal Separation Processes, Technische Universität Hamburg-Harburg, Hamburg, Germany

INTRODUCTION
Carotenoids are natural pigments that give yellow, orange, or red colour to fruits, vegetables
and plants, and helps preventing cardiovascular diseases and cancer [1]. Among carotenoids,
lycopene from tomato is of special interest because of its particular functional properties. In
literature there are reports on SCFE of tomato using SC CO2 as solvent at a laboratory scale
[2,4], but none at pilot-plant or larger scale. Most works use tomato processing by-products as
raw materials, and grinding and drying as pre-treatments.
The objective of this work was to study the scaling-up of SCFE of tomato pellets using SC CO2
at high pressure, considering a one-pass, screening unit and three pilot plants with solvent-
recycling capabilities and different sizes.

MATERIALS & METHODS

We extracted dehydrated commercial tomato flakes containing 6% water. These flakes were
pelletized in two different samples (MP1 and MP2). Moisture of tomato pellets was measured
gravimetrically by drying samples in a convection oven at 105 °C to constant weight (12-17 h).
Oleoresin content was measured by Sohxlet extraction (24 h) using hexane. Lycopene content
in acetone-extracted and saponified pellet samples was measured by HPCL. Screening studies
were done in a computer controlled one-pass laboratory unit (LU) extracting 40-g samples of
MP1 with 12 g/min of CO2 at 40 or 60 °C and 30 or 50 MPa. Pilot plant extractions were all
carried out at 60 °C and 50 MPa (best conditions in screening extractions) in three different
sizes plants. Pilot plant PP1 (Chile) extracted 380-g samples of MP1 (Ub = 760 kg/m3) in 500-
cm3 extraction vessel using 90 g/min of CO2. Pilot plant PP2 (Germany) extracted 340-g of
samples of MP1 (Ub = 708 kg/m3) placed in a 480-cm3 basket of a 1.3-dm3 pilot plant (PP2)
using 288 g/min of CO2. Pilot plant PP3 extracted 1260-g samples of MP2 (Ub = 553 kg/m3)
placed in the 2.28-dm3 basket of a 4-dm3 using using 156 g/min of CO2. Extraction yield was
expressed as percent oleoresin recovered of total available in the substrate.

RESULTS & DISCUSSION

Tomato pellets MP1 and MP2 had 3.4-4.0 mm in diameter and 5.0-5.5 mm in length.
Pelletization of tomato flakes (Ub = 196 kg/m3) increased bulk density of the substrate from
2.4-fold in the case of MP2 to 4.1-fold for MP1. Oleoresin contents were 1.16 in MP1 and
1.47% in MP2. Main difference between pellet samples was in moisture, which was 18%
(w.b.) in sample MP1, and 3.7% (w.b.) in sample MP2. In screening studies, extraction yields
increased with both temperature and pressure. Indeed extraction yield was 2.4% (40 °C and 30
MPa), 8.3% (60 °C and 30 MPa), 17.8% (40 °C and 50 MPa), and 25.1% (60 °C and 50 MPa),
meaning an average improvement of 2.4 times when increasing temperature from 40 to 60 °C,

11th International Congress on Engineering and Food - Athens, Greece, 2011 1715
and a 5.2-fold improvement (average) when increasing pressure from 30 to 50 MPa. Figure 1
shows a comparison among cumulative extraction curve at 60 ºC and 50 MPa for LU, PP1,
PP2, and PP3. Results were unexpected in that the initial slopes of the curves varied widely
depending on the extraction plant, being the same (and highest) in PP1 and PP3, and lowest in
PP2. When cumulative extraction curves plotted as oleoresin yield versus specific CO2
consumption do not coincide for different superficial CO2 velocities, it can be claimed that the
process is not controlled by solubility phenomena but rather by inner mass transfer phenomena.

Figure 1. Kinetics of the SCFE of tomato pellets at 60 °C and 50 MPa. Comparison of the extraction
yield in function of time for three different-sized pilot plants and laboratory scale.

CONCLUSION
In screening studies, oleoresin yield increased more than twice when increasing temperature
from 40 to 60 °C, and more than five times when increasing pressure from 30 to 50 MPa. The
highest yield in LU was 25.1% at 60 °C and 50 MPa. Scaling-up experiments under these
conditions produced unexpected results. Extraction yield in a 500-cm3 pilot plant decreased to
17.6% but increased to 30.7% in 4-dm3 pilot plant. These experiments used two different
substrates, and we believe differences were due to changes in initial moisture and bulk density
between them. Packing high-moisture pellets densely possible resulted in agglomeration of the
substrate and undesirable channelling within the packed bed.
Acknowledgements. This work was funded by Fondecyt (project 108-0211) from Chile.

REFERENCES
[1] Perera, C. O. & Yen, G. M. 2007. Functional properties of carotenoids in human health. International
Journal of Food Properties, 10(2), 201-230. [2] Sabio, E., Lozano, M., de Espinosa, V. M., Mendes, R. L.,
Pereira, A. P., Palavra, A. F. & Coelho, J.A. 2003. Lycopene and beta-carotene extraction from tomato
processing waste using supercritical CO2. Industrial & Engineering Chemistry Research, 42(25), 6641-
6646. [3] Topal, U., Sasaki, M., Goto, M. & Hayakawa, K. 2006. Extraction of lycopene from tomato
skin with supercritical carbon dioxide: Effect of operating conditions and solubility analysis. Journal of
Agricultural and Food Chemistry, 54(15), 5604-5610. [4] Rozzi, N. L., Singh, R. K., Vierling, R. A. &
Watkins, B. A. 2002. Supercritical fluid extraction of lycopene from tomato processing byproducts.
Journal of Agricultural and Food Chemistry, 50(9), 2638-2643.

1716
 Supercritical extraction of astaxanthin from H. pluvialis using ethanol-modified CO2.

Experiments and modelling.

A. Bustamante, P. Roberts,1 R. Aravena, J.M. del Valle2

1
Departamento de Ciencia y Tecnología Química de los Alimentos, Facultad Ciencias Químicas y
Farmacéuticas, Universidad de Chile, Santiago, Chile. (proberts@uchile.cl)
2
Departamento de Ingeniería Química y Bioprocesos, Pontificia Universidad Católica de Chile,
Santiago, Chile.(delvalle@ing.puc.cl)

INTRODUCTION
Haematococcus pluvialis is an interesting alterative for obtain astaxanthin from a natural
source. Astaxanthin can be extracted from H. pluvialis using SuperCritical (SC) carbon dioxide
(CO2). Several researchers studied SC CO2 extraction of astaxanthin from H. pluvialis
reporting high yield, and high concentrations of astaxanthin in extracts depending on operating
conditions [1, 2]. With the objective of obtaining an astaxanthin rich extract, we used response
surface (RS) analysis to evaluate the effect of extraction temperature, extraction pressure and
ethanol concentration on astaxanthin recovery in 5-h SC CO2 extraction experiments. Results
were not as expected. We speculated that anomalies were due to kinetic effects during
extraction. Thus, the main objective of this work was measuring and modelling kinetic curves
of astaxanthin extraction under selected conditions to evaluate the effect of extraction time on
astaxanthin recovery.

MATERIALS & METHODS

Treatments. H. pluvialis were extracted in a one-pass laboratory SCE device. Extractions were
carried out at 313, 328, or 343 K, and 30, 42.5, or 55 MPa, using 4.5 g/min of pure or ethanol-
modified (4 or 8% v/v) CO2 during 5 h.
Experimental design. RS analysis was used to evaluate the effects of three independent
variables on astaxanthin recovery.
Modeling. Extraction curves were modeled using the model of Sovova [3]. This model
considers a tissue containing a fraction of broken cells and a remainder of intact cells. The
mass transfer occurs from intact to broken cells and from broken cells to SC CO2.

RESULTS & DISCUSSION

Response Surface Analysis. Depending on extraction conditions and ethanol added,
astaxanthin recoveries ranged 43-82.8%. Results indicate no interactions between independent
variables on astaxanthin recovery. The increase in pressure had a large positive effect in the
astaxanthin recovery, but it was lower that reported by others [1, 2]. On the other hand, the
astaxanthin recovery decrease as a result of an increase in temperature is opposite observation
of others [1, 2]. Besides, the use of ethanol as co-solvent improves astaxanthin recovery, as
expected.
Kinetic curves of extraction and modelling. The kinetics curves (Figure 1) shows that
extraction rate is very slow after 2 h (180 g CO2/g microalga), which indicates that longer
extraction times do not contribute to astaxanthin recovery to a great extent.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1717
Regarding the solubility of astaxanthin in pure CO2, model shows an increase with density, as
expected. However, the solubility of astaxanthin in blend of CO2 and ethanol is slightly lower
that in pure CO2, which was not expected. On the other hand, the others parameters that
characterize the sorcion isotherm were effected for use ethanol.
With respect to mass transfer parameters, the fraction of broken cells (58%) was considered
constant because it depends only of the raw material. On the other hand, in all cases external
mass transfer coefficient is much larger than external mass transfer coefficient as reported by
Sovová [3]. Besides, added ethanol improved the internal mass transfer, being the internal mass
transfer coefficient with co-solvent 10 times higher than pure CO2.
Considering all results of kinetic studies, it seems that the improvement in astaxanthin recovery
when adding ethanol is due to its large positive effect over the mass transfer process rather than
its effect in equilibrium.

Figure 1. Kinetic curves of astaxanthin extraction from H. Pluvialis

CONCLUSIONS
The results show that in the experimental region the pressure and add ethanol has a positive,
and temperature has a negative effect. Besides, model of Sovova [3] fitted well astaxanthin
extraction curves using both pure and ethanol-modified CO2. Model parameters suggest that
the positive effect of added ethanol is due to enhancement of the mass transfer from unbroken
cells.

REFERENCES
[1] Machmudah S., Shotipruk A., Goto M., Sasaki M. & Hirose T. 2006. Extraction of astaxanthin from
Haematococcus pluvialis using supercritical CO2 and ethanol as entrainer. Industrial Engineering
Chemistry Research, 45, 3652-3657.
[2] Thana P., Machmudah S., Goto M., Sasaki M., Pavasant P. & Shotipruk A. 2008. Response surface
methodology to supercritical carbon dioxide extraction of astaxanthin from Haematococcus pluvialis.
Bioresource Technology, 99(8), 3110-3115.
[3]Sovova H. 2005. Mathematical model for supercritical fluid extraction of natural products and
extraction curve evaluation. Journal of Supercritical Fluids, 33, 35-52.

1718
 Supercritical carbon dioxide extraction and fractionation of rapeseed cake oil

Edgar Uquichea, Katherine Salazara, Ximena Ficaa, José Manuel del Valleb

a
Universidad de La Frontera (UFRO), Temuco, Chile, (euquiche@ufro.cl)
b
Pontificia Universidad Católica de Chile, Santiago, Chile, (delvalle@ing.puc.cl)

INTRODUCTION
Rapeseed (Brassica napus) press cake rich in lipids and minor lipids such as phytosterols,
tocopherols, and carotenoids of great functional value as natural antioxidants. Fractionation
effects during extraction that may increase the concentration of these minor lipids in the oil are
desirable, and this work explored the possibility of using SuperCritical carbon dioxide (SC-
CO2) to achieve fractionation of press cake rapeseed oil. The objective of this study was to
identify the effects of various combinations of medium to high pressures and temperatures and
short times on the yield of oil and the concentration of minor lipid in fractions of SC-CO2-
extracted cold-pressed rapeseed cake, with the aim of separating fraction enriched in minor
lipids.

MATERIALS & METHODS

Supercritical extraction was carried out in a Spe-ed SFE unit (Applied Separations, Allentown,
PA). Milled cold-pressed rapeseed cake samples were placed in a 50 cm3 vessel and extracted
with CO2. Extractions were carried out at 30, 35, 40, 45, or 50 MPa and at 40 or 80 ºC, at
solvent flow corresponding to superficial velocity of 1 mm/s. Recovered oil was assessed
gravimetrically by difference with cleaned and dried vials, and cumulative oil yields were
calculated. Oil in vials was dissolved in chloroform p.a. and flashed to 50 cm3 in volumetric
flask prior quantification of sterols, tocopherols, and carotenoids by UV spectrophotometry.

RESULTS & DISCUSSION

Cumulative oil yield increased with extraction pressure, and with extraction temperature at •40
MPa, but was lower at 80 than 40 ºC at ”35 MPa, which is consistent with a solubility-
controlled process and a crossover pressure between 35 and 40 MPa. Figure 1 plots cumulative
oil yield versus specific solvent consumption to unveil the relationship between extraction rate
and oil solubility based on the correlation proposed by del Valle et al. (submitted) for the
solubility of vegetable oils in SC-CO2. In Figure 1, the 45º line by the origin represents
extractions carried out under the solubility-controlled conditions hypothesized for the initial
stages of the SC-CO2 extraction process. It appear as if extractions are solubility-controlled as
long as the total amount of oil extracted is limited to ca. 25-40% of the total (40-60 g oil/kg dry
substrate), and that the correlation of del Valle et al. (submitted) provides good estimates of
rapeseed oil solubility within the experimental region. Besides a solubility-controlled initial
period, SC-CO2 extraction curves consider subsequent periods where extraction rate is
controlled by external mass transfer mechanisms, inner mass transfer mechanisms, and
desorption mechanisms that are markedly dependent on the substrate pretreatment and particle
size (Güçlü-Üstündag & Temelli, 2004).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1719
Figure 1. Cumulative oil yield versus specific solvent consumption at ( , ) 30 MPa, ( , ) 35 MPa,
( , ) 40 MPa, ( , ) 45 MPa, or ( , ) 50 MPa, ( , , , , ); 40 ºC or ( , , , , ) 80 ºC.

The weight and concentration of minor lipids (sterols, tocopherols, carotenoids) in oil fractions
collected during the first 60 min of extraction were recorded and analyzed. Differences in
solubility between the oil and minor lipids explained fractionation effects that were small for
tocopherols. Unlike tocopherols, that are more soluble in SC-CO2 than the oil, sterols and
carotenoids are less soluble than the oil, and their concentration increased in the later stages of
the extraction process, particularly at •40 MPa, when there was no enough oil to saturate the
CO2 phase. Consequently, this study suggests that SC-CO2 extraction can be used to isolate
vegetable oil fractions having increased functional value.

CONCLUSIONS
SC-CO2 extraction of rapeseed oil appears to be a solubility-controlled partially dependent on
residual oil concentration in the prepressed seeds. Differences in solubility between the oil and
minor lipids (sterols, tocopherols, carotenoids) also explain fractionation effects. These
fractionation effects are small for tocopherols that are more soluble in SC-CO2 than the oil. On
the other hand, there is an increase in concentration of sterols and carotenoids in the later
stages of the extraction process, when there is no enough oil to saturate the CO2 phase, because
these two components are less soluble in SC-CO2 than the oil.

REFERENCES
[1] del Valle, J.M., Uquiche, E.L. & de la Fuente J.C. A refined equation for predicting the solubility of
vegetable oils in high-pressure CO2. The Journal of Supercritical Fluids (submitted).
[2] Güçlü-Üstünda÷, Ö. & Temelli, F. 2004. Correlating the solubility behavior of minor lipid
components in supercritical carbon dioxide. The Journal of Supercritical Fluids, 31, 235–253.

1720
 Characterization of Novel Cholesterol Esterase from Trichoderma sp. AS59 with High

Ability to Synthesize Steryl Esters

Atsushi Maedaa, Norihumi Hashitania, Takayuki Mizunoa, Masanori Bunyaa

a
Faculty of Engineering, Tokushima Bunri University, 1314-1 Shido, Sanuki 769-2193, Japan
(maeda@fe.bunri-u.ac.jp)

INTRODUCTION
The structures of phytosterols that are naturally occurring in plants are very similar to that of
cholesterol, and they are effective in inhibiting the absorption of cholesterol in the small
intestine. Thus, phytosterols have promise as a functional food material. However, they have
not been actively used as material for low solubility in both oil and water.
We found a novel cholesterol esterase to synthesize steryl ester from the culture filtrate of a
fungal strain Trichoderma sp. AS59 isolated from soil [1]. In this study, we examined the
ability of the enzyme to synthesize steryl esters from sterol and free fatty acids of varying chain
lengths.
If the ability of the synthesis of steryl ester is similar to that of the hydrolysis, it is possible to
synthesis the useful sterol esters as a functional food material. This leads to further widen the
application of plant sterol to food, and it is still challenging work.

MATERIALS & METHODS

Cholesterol esterase from Trichoderma sp. AS59 was purified from a 3-day liquid culture
according to the method already reported [5]. The substrate solution 30 mL of hexane
including 100 mmol/L of free fatty acids and saturated sterol. Then, the esterification was
examined by mixing 30 mL of the substrate solution with lyophilized enzyme and a certain
amount of water in 50 mL glass vial. A mixture was shaken at 120 rpm in a 27oC water bath
for 120 h. The products and the remaining reactants were determined by gas chromatography.

RESULTS & DISCUSSION

We measured the saturated concentration of cholesterol in hexane by GC, and it was about 25
mmol/L. Figure 1 shows the effect of chain length of fatty acid on cholesterol esterification.
40

30
Yield [%]

20

10

0
3 4 6 8 10 12 14 16 18
Chain length of saturated fatty acid

Figure 1. Effect of chain length of FFAs on cholesterol esterification.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1721
This indicates that all the medium- and long-chain FFAs used were successfully employed in
ester synthesis, whereas the short-chain FFAs were unavailable for this reaction. In the
previous study [1], the short chain fatty acid cholesteryl esters (especially cholesteryl butyrate)
were available for hydrolysis. It is clear that the fatty acid specificity in ester synthesis is
entirely different from that in hydrolysis.

6
Yield [%]

0
0 20 40 60 80 100 120 140

Time [h]

Figure 2. Time course of esterification of stigmasterol with palmitic acid. A substrate mixture
containing 100 mmol/L of palmitic acid and 7.0 mmol/L of stigmasterol was combined with
100 U of the enzyme dissolved in 20 PL of 0.2M phosphate buffer (pH 7.0) at 27oC.

Figure 2 shows the time course of the esterification of stigmasterol with palmitic acid. The
yield of stigmasteryl palmitate was reached to 7%. However, extending the reaction time even
further didn’t also improve the yield. Because the saturated concentration of stigmasterol in
hexane measured by GC was about 5.6 mmol/L, the chemical equilibrium was established at
the time of synthesis of some stigmasteryl palmitate. In these conditions, the enzyme turned
into paste and clung to the inside of the vial. It is easy to separate the enzyme and the solvent,
then the reaction rate may be recovered and the mass of steryl ester may increase by replacing
the substrate solution.

CONCLUSION
All the medium- and long-chain FFAs used are successfully employed in cholesterol ester
synthesis in large scale such as 30 mL of solvent. We were also able to synthesize stigmasterol
palmitate as the plant sterol ester. These results could pave the way large-scale synthesis of
sterol ester and indicate the potential utility of the enzyme in the food industry.

REFERENCES
[1] Maeda A., Mizuno T., Bunya M., Sugihara S., Nakayama D., & Tsunasawa S. 2008. Characterization
of novel cholesterol esterase from Trichoderma sp. AS59 with high ability to synthesize steryl esters.
J Biosci Bioeng, 105, 341-349.

1722
Recovery of an antibacterial peptide fraction from snow crab by-products hydrolysate by

electrodialysis with ultrafiltration membranes

Alain Doyena,b, Linda Sauciera,c, Lucie Beaulieua,d, Yves Pouliota,b, Monica Araya-Fariasa,b, Laurent
Bazineta,b

a
Institute of Nutraceutical and Functional Foods (INAF), Université Laval, Québec, Canada.
laurent.bazinet@fsaa.ulaval.ca
b
Department of Food Science and Nutrition, Université Laval, Québec, Canada
c
Department of Animal Sciences, Université Laval, Québec, Canada
d
Department of Biology, Chemistry and Geography, Université du Québec à Rimouski (UQAR),
Rimouski, Canada

INTRODUCTION
By-products and surpluses from food industries could generate peptides with great interest for
food formulation. Hence, the separation and concentration of bioactive peptides from natural
sources becomes increasingly attractive to the food industry [1]. Recently, a snow crab by-
products hydrolysate showed antibacterial activity [2]. This activity was due to an antibacterial
peptide of about 800 Da [2]. However, the antibacterial activity was only detected at high
peptide concentration. Consequently, peptide hydrolysate has to be fractionated to obtain
peptides in a more purified form. Electrodialysis with ultrafiltration membranes (EDUF),
developed and patented in 2005 [3], allowed separation of molecules from a complex mixture
according to their charge and their molecular weights. EDUF had already successfully
demonstrated the recovery of interesting peptide fractions [4, 5]. Hence, the aim of this work
was to recover and concentrate the active antibacterial fraction.

MATERIALS & METHODS

The electrodialysis cell used for this experiment was a MP type cell with one cation exchange
membrane (CEM) membrane, one anion exchange membrane (AEM) and two UFM with
MWCO of 50 kDa or 20 kDa. The electrodialysis configuration was divided into four
compartments. Two of them containing 1.5L of KCl solution for the recovery and
concentration of peptides (compartments KCl1 and KCl2 for the recovery of anionic and
cationic peptides respectively) . One compartment containing the electrode rinsing solution and
another one for the feed solution (snow crab by-products hydrolysate). Electroseparation was
performed in batch process using electrical field strengths of 2 V/cm or 14 V/cm during six
hours. Afterwards, the evolution of peptide concentration in the KCl compartments were
determined by BCA protein assays and peptide molecular weight and distribution in the KCl
compartment was determined by LC-MS and the KCl fractions were tested on Escherichia coli
ATCC 25922 and Listeria innocua HPB 13.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1723
RESULTS & DISCUSSION
In the KCl1 compartment, the highest peptide transport rate was obtained at 14 V/cm with
UFM MWCO of 50 kDa with a value of 32.4 g/m2.h. At 2 V/cm and whatever the UFM
MWCO, similar values of peptide transport rates were calculated. For the KCl2 compartment
and contrary to KCl1 compartment, the peptide transport rate and peptide percentages in KCl
powders remained low during the electroseparation. As observed for the KCl1 compartment,
the important electrical field strength applied in the system (14 V/cm) combined to a large
UFM MWCO (50 kDa) allowed the highest peptide migration. With respect to the peptide
molecular weight and distribution, in the KCl1 and KCl2 compartments after EDUF, whatever
the electric field strength and UFM MWCO, the 300-600 Da molecular weights fraction was
the most abundant with an average of about 94% and was also the one with the highest
migration rate. The 800-900 Da molecular weight range peptide (peptide which demonstrated
antibacterial properties in the initial snow crab by-products hydrolysate) was only detected in
the KCl1 compartment after electroseparation performed at 14V/cm with UFM MWCO of 50
kDa; this molecular weight range represented a migration rate of about 30%. With respect to
the antibacterial activities, only the KCl1 fraction at 50g of peptide/L and recovered after
EDUF separation at 14 V/cm with UFM MWCO of 50 kDa showed antibacterial activity on
Escherichia coli ATCC 25922 and Listeria innocua HPB 13with inhibition diameter of 4.0
mm. The initial snow crab by-product hydrolysate at the same concentration (50 g of
peptide/L) did not exhibit antibacterial activity.

CONCLUSION
These results demonstrated the selectivity of the EDUF process concerning the migration and
the concentration of specific peptide fractions depending on the process parameters (electrical
field strength and UFM MWCO). Finally, different experiments to determine amino acid
sequence of the antibacterial peptide in the EDUF antibacterial fraction (KCl1 recovered after
EDUF separation performed at 14V/cm with UFM MWCO of 50 kDa) will be carried-out in
the near future.

REFERENCES
[1] H. Korhonen, A. Pihlanto. 2003. Food-derived Bioactive Peptides – Opportunities for Designing
Future Foods. Current Pharmaceutical Design, 9 (16), 1297-1308.
[2] L. Beaulieu, J. Thibodeau, M. Desbiens, R. Saint-Louis, C. Zatylny-Gaudin, S. Thibault. 2010.
Evidence of Antibacterial Activities in Peptide Fractions Originating from Snow Crab (Chionoecetes
opilio) By-Products. Probiotics and Antimicrobial Proteins, 2 (3), 1-13.
[3] L. Bazinet, J. Amiot, J.F. Poulin, A. Tremblay, D. Labbé. 2005. Process and system for separation of
organic charged compounds. Brevet PCT/CA2005/000337.
[4] J.F. Poulin, J. Amiot, L. Bazinet. 2006. Simultaneous separation of acid and basic bioactive peptides
by electrodialysis with ultrafiltration membrane. Journal of Biotechnology, 123 (3), 314-328
[5] L. Firdaous, P. Dhulster, J. Amiot, A. Doyen, F. Lutin, L.P. Vézina, L. Bazinet. 2010. Investigation of
the large-scale bioseparation of an antihypertensive peptide from alfalfa white protein hydrolysate by
an electromembrane process. Journal of Membrane Science, 355 (1-2), 175-181.

1724
 Prospection of bacterial endophytes isolated from Baru (Dipteryx alata Vog.) as a

potential source of bioactive compounds

Gustavo Molinaa, Ana Paula Dionísioa, Mariana Recco Pimentela, Gisele Tokie Makitaa, Renato Correia
dos Reisa, Gláucia Maria Pastorea

a
Department of Food Science, School of Food Engineering, University of Campinas (UNICAMP).
Campinas, São Paulo, Brazil (gustavomolinagm@gmail.com)

INTRODUCTION
The term “endophytes” includes a suite of microorganisms that grow intra and/or
intercelullarly in the tissues of higher plants without causing over symptoms on the plants in
which they live. These micro-organisms represents a potential source of novel natural products
for medicinal, agricultural and industrial uses, such as antibiotics, anticancer agents, biological
control agents, and other bioactive compounds [1, 2].
Endophytes provide a broad variety of bioactive secondary metabolites with unique structure,
including alkaloids, benzopyranones, chinones, flavonoids, phenolic acids, quinones, steroids,
terpenoids, tetralones, xanthones, and others [3]. Such bioactive metabolites find wide-ranging
application as agrochemicals, antibiotics, immunosuppressants, antiparasitics, antioxidants, and
anticancer agents [4].
The aim of the present work was to investigate the biotechnological potential of bacterial
endophytes isolated from Baru (Dipteryx alata Vog.). Accordingly, the antimicrobial activity,
the enzymatic profile and the biotransformation of Į-pinene were evaluated.

MATERIALS & METHODS

In the biotransformation process, the bacterial culture was grown and the concentrated biomass
was placed in a 250 mL conical flask containing 50 mL of mineral medium. Biotransformation
was started by adding 0,5% (v/v) of Į-pinene, and the flasks were incubated in rotary shaker at
30 ºC and 150 rpm, and samples were monitored by gas chromatography until 96h. To access
antimicrobial activity, one full loop of the 24h old culture was transferred to a 50 mL
Erlenmeyer flask containing 10 mL of liquid YM medium. After 8 days of incubation at 30 °C
and 150 rpm, the biomass was recovered by vacuum filtration with membrane 0.22 ȝm
(Millex® - Millipore) to obtain the bacterial extract, which was tested against pathogenic
cultures by the disc diffusion method. The isolated strains were also tested for their ability to
produce extracellular enzymes that degrade culture medium containing starch, protein and
olive oil, for a screening of amylase, protease and lipase enzymes production, respectively.

RESULTS & DISCUSSION

A total of 17 bacteria strains were isolated from Baru and identified in the present work.
Screening of the antimicrobial activity of endophytic extracts revealed a considerable activity
against the pathogenic cultures tested. Most of the extracts inhibited the growth of Candida
albicans, Escherichia coli and Sthaphylococcus aureus.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1725
Furthermore, it was observed that the vast majority of bacteria showed a high level protease
activity, the lipase and amylase profile was detected in most isolates but with less intensity.
The preliminary results obtained from the biotransformation process showed that 2 bacterial
strains bioconverted Į-pinene into verbenol (85% similarity in MS results). The bioconversion
occurred based on the biochemical reaction of hydroxylation of Į-pinene, and this reaction was
reported in some articles [5, 6].

CONCLUSION
This paper represents the use of bacterial endophytes isolated from the Cerrado brazilian biome
and demonstrates a partial use of these microorganisms in biotechnological processes and their
potential as source of bioactive compounds.

REFERENCES
[1] Li J., Zhao G.Z., Chen H.H., Wang H.B., Qin S., Zhu W.Y., Xu L.H., Jiang C.L. & Li W.J. 2008.
Antitumour and Antimicrobial Activities of Endophytic Streptomycetes from Pharmaceutical Plants
in Rainforest. Letters in Applied Microbiology, 47, 574–580.
[2] Tan R.X. & Zou W.X. 2001. Endophytes: a Rich Source of Functional Metabolites. Natural Product
Reports, 18, 448–459.
[3] Kogel K.H., Franken P. & Huckelhoven R. 2006. Endophyte or Parasite — What Decides? Current
Opinion in Plant Biology, 9, 358–363.
[4] Strobel G.A. 2003. Endophytes as Sources of Bioactive Products. Microbes and Infection, 5, 535–
544.
[5] Maróstica Jr M.R., Mota N.O., Baudet N. & Pastore G.M. 2007. Fungal Biotransformation of
Monoterpenes Found in Agro-Industrial Residues from Orange and Pulp Industries into Aroma
Compounds: Screening using Solid Phase Microextraction. Food Science Biotechnology, 16.
[6] Agrawal R., Deepika N.U.A. & Joseph R. 1999. Strain Improvement of Aspergillus sp. and
Penicillium sp. by Induced Mutation for Biotransformaiton of Į-pinene to Verbenol. Biotechnology
and Bioengineering, 63, 249-252.

1726
 Biotransformation of R-(+)- and S-(௅)-limonene by Fusarium oxysporum

Gustavo Molinaa, Renata Lino da Costaa, Ana Paula Dionísioa, Juliano Lemos Bicasb, Gláucia Maria
Pastorea

a
Department of Food Science, School of Food Engineering, University of Campinas (UNICAMP).
Campinas, São Paulo, Brazil (gustavomolinagm@gmail.com)
b
Cap, Federal University of São João Del Rey. Ouro Branco, Minas Gerais, Brazil

INTRODUCTION
In the past few years much work has been done on the biotransformation of limonene, an
inexpensive hydrocarbon monoterpene, which is one of the most widely distributed terpene in
nature. As its chemical structure is similar to that of many oxygenated monoterpenoids
presenting a pleasant fragrance, e.g. perillyl alcohol, carveol, carvone, menthol and Į-terpineol,
it may be used as a precursor in the synthesis of these flavor compounds [1].
Recently, the strain F. oxysporum 152b has been selected based on its high production of
extracellular alkaline lipase [2]. Simultaneously, other publications have also described the
biotransformation of R-(+)-limonene into R-(+)-Į-terpineol catalysed by the same strain [3,4].
The main characteristics of this conversion were described in recent papers [5] but no
information deals with the bioconversion of its isomer, S-(௅)-limonene.

MATERIALS & METHODS

A 72 h culture grown on agar in a Petri dish was divided amongst 250 mL conical flasks
containing 50 mL of YM medium (10 g.L-1 glucose, 5 g.L-1 peptone, 3 g.L-1 yeast extract, 3
g.L-1 malt extract, pH 6.7) and homogenized under sterile conditions using an Ultra-Turrax®
T18 (Ika, Wilmington, NC, USA) until complete disruption of the solid matter. After 72 h
incubation at 30 °C/150 rpm, the cell mass was concentrated by vacuum filtration using a
Buchner funnel with Whatman n° 1 filter paper. The concentrated biomass was placed in a 250
mL conical flask containing 50mL of mineral medium. Biotransformation was started by
adding 0,5% (v/v) of S-(௅)-limonene, the flasks were incubated in rotary shaker at 30 ºC and
150 rpm and samples were monitored by gas chromatography until 96h.

RESULTS & DISCUSSION

In former studies, Bicas et al [3] optimised the medium composition and the culture conditions
involved in the biotransformation of R-(+)-limonene by Fusarium oxysporum. In sequence,
Bicas et al [5] described the production of almost 4 g.L-1 of Į-terpineol after 48 h of
fermentation, which remained stable until reaching 96 h.
However, the biotransformation of S-(-)-limonene led to the production of 1.2 g.L-1 limonene-
1,2-diol, under non optimised conditions. This is a result from the ring double bond
epoxidation of limonene, followed by the corresponding diol formation. However, the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1727
production of the intermediate limonene-1,2-epoxide was not detected, indicating that the
reaction proceeds mainly for the diol production.
It was proven that the enzyme responsible for the production of Į-terpineol by the
biotransformation of R-(+)-limonene was enantioselective and enantiospecific, has an
intracellular nature and acts in anaerobic conditions. In the present study, it was observed that
the production of limonene-1,2-diol from S-(௅)-limonene has also an intracellular nature but it
was highly influenced by an aerobic system and seemed to be cofactor dependent, considering
that the product was not detected with an anaerobic biotransformation.

CONCLUSION
The results collected encourage further studies with this biocatalyst and the characterization of
its enzyme system, involved in the biotransformation of S-(௅)-limonene, considering the
concentration of limonene-1,2-diol achieved. Moreover, the optimization of culture conditions
could enhance its production and the application of other terpenes as substrate is of great
importance and could be source of novel aroma compounds with industrial interest.

REFERENCES
[1] Bicas J.L., Dionísio A.P. & Pastore G.M. 2009. Bio-oxidation of Terpenes: An Approach for the
Flavor Industry. Chemical Reviews, 109, 4518-4531.
[2] Prazeres J.N., Cruz J.A.B. & Pastore G.M. 2006. Characterization of Alkaline Lipase from Fusarium
oxysporum and the Effect of Different Surfactants and Detergents on the Enzyme Activity. Brazilian
Journal of Microbiology, 37, 505–509.
[3] Bicas J.L., Barros F.F.C., Wagner R., Godoy H.T. & Pastore G.M. 2008. Optimization of R-(+)-Į-
terpineol Production by the Biotransformation of R-(+)-Limonene. Journal of Industrial Microbiology
and Biotechnology, 35, 1061-1070.
[4] Maróstica Jr M.R. & Pastore G.M. 2007. Production of R-(+)-Į-Terpineol by the Biotransformation of
Limonene from Orange Essential Oil, using Cassava Waste Water as Medium. Food Science, 101,
345-350.
[5] Bicas J.L., Quadros C.P., Neri-Numa I.A. & Pastore G.M. 2010. Integrated process for co-production
of alkaline lipase and R-(+)-a-terpineol by Fusarium oxysporum. Food Chemistry, 120, 452–456.

1728
 Pulsed Light Decontamination of Vegetables and Fruits

Gianpiero Pataro a*, Giorgio Donsì a,b, Giovanna Ferrari a,b

a
Department of Industrial Engineering, University of Salerno, Fisciano, Italy (gpataro@unisa.it)
b
ProdAl scarl, Fisciano, Italy (info@prodalricerche.it)

INTRODUCTION
Pulsed light (PL) is an emerging technology which has considerable potential as an alternative
to thermal and chemical methods for rapid and effective inactivation of microorganisms on
food surfaces. The technique involves the use of intense short duration light pulses of a wide
broad spectrum light (100-1100 nm) [1]. A significant number of publications on the subject
have documented the ability of PL to inactivate microbial species spread on agar surfaces [2],
while only few publications are dealing with the PL decontamination of food products [3].
In this work the ability of PL treatments for surface decontamination of vegetables and fruits
by killing native microflora was investigated.

MATERIALS & METHODS

Samples of three varieties of tomatoes (Micron, BNC8015 and AA7033) and two varieties of
peaches (Lucia and Duceur) were exposed to PL treatments at different energy doses (35-250
J/cm2/side). PL flashes (3 pulses/s, pulse width 360 Ps, 0.39 J/cm2/pulse) were generated by
SteriPulse®-XL 3000 Pulsed light Sterilization System (Xenon Corp., Wilmington, Mass.,
USA). The lethality of PL treatments was assessed by the total aerobic mesophilic count and
yeast and mould population.

RESULTS & DISCUSSION

The results reported in Table 1 show that the reduction of the native microflora is progressively
increased with increasing the energy incident on the surface of each product. However, no
complete inactivation was detected even when the higher energy dose was applied.
At fixed energy dose applied the effectiveness of the treatment depended on the PL sensitivity
of the microbial population living on the surface of each variety of product. The native
microflora present on the surface of tomatoes of the variety Micron highlighted the greater
sensitivity to light pulses followed by the one living on the surface of the tomatoes of the
variety BNC8015 and AA7033. Between the two variety of peaches tested microbial population
living on the surface of Lucia variety showed the higher sensitivity to light pulses.
Thermal damages were visible especially on the surface of the peaches when treatments of
higher energy dose were applied. Therefore, multi-step treatment in which each side of the
product was exposed several times to light pulses of low energy dose (31.4 J/cm2/side) per step
made it possible to achieve the same lethal effect along with the minimum thermal damage
(data not shown).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1729
Table 1. Microbial counts of total plate counts and yeast and mould in fresh and PL treated tomatoes and
peaches.
Product Product Variety Energy dose* Aerobic mesophilic count Yeast and Mould
J/cm2/side Log-CFU/mL Log-CFU/mL
Tomato Micron 0 3.16 2.48
6 2.70 1.89
18 1.51 0.90
35 0.78 0.30
BNC8015 0 2.48 2.42
6 2.49 2.17
18 1.62 1.93
35 0.85 1.57
AA7033 0 2.00 2.54
6 1.95 2.50
18 1.95 2.45
35 1.00 1.36
Peaches Lucia 0 3,42 2,89
21 3,22 2,81
31 2,22 2,35
63 1,91 1,80
126 1,51 1,10
252 0,92 0,69
Duceur 0 3,98 3,22
21 3,68 3,01
31 2,88 2,55
63 2,23 2,44
126 1,59 1,39
252 0,82 0,69
* Two sides per fruit were considered

CONCLUSION
Our results suggest that PL treatment operated under optimal conditions may constitute an
effective method for the surface decontamination of fruit and vegetable products.

REFERENCES
[1] Krishnamurthy K., Tewari J.C., Irudayaraj J. & Demirci A. 2008. Microscopic and spectroscopic
evaluation of inactivation of Staphylococcus aureus by pulsed UV light and infrared heating. Food
and Bioprocess Technology. doi:10.1007/s11947-008-0084-8.
[2] Elmnasser N., Guillou S., Leroi F., Orange N., Bakhrouf A. & Federighi M. 2007. Pulsed-light system
as a novel food decontamination technology: A review. Canadian Journal of Microbiology, 53, 813–
821.
[3] Gómez-López, V.M., Devlieghere, F., Bonduelle, V. & Debevere, J. 2005. Intense light pulses
decontamination of minimally processed vegetables and their shelf-life. International Journal of Food
Microbiology, 103, 79-89.

1730
 Shelf life extension of fresh-cut fruit by UV-light exposure

Lara Manzocco, Sara Da Pieve, Ingrid Bartolomeoli, Michela Maifreni

Dipartimento di Scienze degli Alimenti Università degli Studi di Udine, Udine, Italy
(lara.manzocco@uniud.it)

INTRODUCTION
Fresh-cut fruit and vegetables are tremendously growing segments in retail establishments due
to increasing consumer demand for fresh, healthy and convenient foods. Fresh-cut processing
is well known to promote a faster degradation. For this reason, new technologies for
maintaining quality while inhibiting undesired microbial growth are increasingly investigated.
Ultraviolet-C (UV-C) treatment from 200 to 280 nm is a powerful non-thermal germicidal
method which has raised a large attention due to easy use and favourable costs of equipments,
energy and maintenance [1]. Moreover, such technology has been reported to promote
enzymatic inactivation in vegetable tissues without forming known toxic or significant non-
toxic by products. The aim of this study was to evaluate the effect of UV-C light on safety and
quality of fresh-cut fruit.

MATERIALS & METHODS

Fruits were cut in cubes, slices or sticks, irradiated with increasing UV-C light doses up to
12000 J/m2 (maximum emission 253.7 nm) and packed. During storage at 6 °C up to 10 days,
samples were analysed for colour, firmness, juice leakage and microbial population.
Microscopic analyses of plant tissues as well as trained sensory panel analysis and preference
analysis were also performed during storage.

RESULTS & DISCUSSION

Microbial populations of fresh-cut samples were significantly reduced by light exposure. UV-C
light treatment leaded to a number of log reductions higher than 2 for total viable count,
enterobacteria and yeasts and moulds. In particular, Table 1 shows the effect of UV-C light
exposure on total viable count of different fresh-cut fruits.

Table 1. Log reductions in total viable count of fresh-cut fruit exposed to 1200J/m2 UV-C light.
Fresh-cut fruit Total viable count Log reductions
Apple slices 3.1 ± 0.3
Melon cubes 2.2 ± 0.2
Pineapple sticks 2.0 ± 0.4
n=15

11th International Congress on Engineering and Food - Athens, Greece, 2011 1731
Total viable counts remained about 2 Log cycles lower than that of the untreated fruit during
the entire storage time at 6 °C.
The decontamination effect of UV-C light is strictly dependent on the fruit matrix considered.
Fruits presenting a flat and smooth surface are generally associated to higher decontamination
efficiency of UV-C radiation. To this regard, intense surface roughness, typical of pineapple
sticks (table 1) is expected to shadow eventual microbial cells, thus impairing the germicidal
effect.
UV-C light treated samples showed not significantly different colour and firmness as compared
to the control. In the case of apple derivatives, this was attributed to complet inactivation of
polyphenoloxidase on cut surface [2] By contrast, leakage of intracellular liquids from UV-C
treated samples was significantly lower, possibly due to the dehydration of a thin surface layer
of fruit tissue upon light exposure [3]. It can be hypothesized that UV-C exposure promotes the
modification of fruit cell structure leading to the rupture of membranes of cells and thus
favouring the progressive dehydration of the sample. The hypothesis of the loss of cell integrity
upon light exposure was also supported by optical microscopy observations performed to
assess the structure changes produced, at a cellular level.
In order to verify the potential effects of UV-C light exposure on the sensory properties of
fresh-cut fruit, samples were submitted to sensory analysis using both trained panel and
consumers. Results indicated UV-C treated samples to have better flavour than the untreated
fruit while no differences were detected in the other sensory descriptors. In addition, UV-C
light treated samples were associated to an off-flavour perception lower than that of the control
during the entire storage, possibly due to lower microbial growth during storage. As suggested
by the specific microbiological criteria for minimally processed fruits and vegetables, a total
viable count of 7 log CFU/g was taken as acceptability limit to estimate shelf life. UV-light
exposure caused a 40 % extension in shelf life.

CONCLUSION
Results indicate that UV-C treatment represents an interesting technology to non-thermally
decontaminate the surface of fresh-cut products, thus extending their shelf life. These effect
can be attributed to: death of spoilage microorganisms physically exposed to the radiation;
prevention of off-flavour formation during storage; formation of a dried protective edible film
inhibiting microbial growth and juice leakage.

REFERENCES
[1] Bintsis, T., Litopoulou-Tzanetaki, E. & Robinson, R. K. 2000. Existing and potential applications of
ultraviolet light in food industry – a critical review. Journal of the Science of Food and Agriculture,
80, 637-645.
[2] Manzocco L., Quarta B. & Dri A. 2009. Polyphenoloxidase inactivation by light exposure in model
systems and apple derivatives. Innovative Food Science and Emerging Technologies, 10, 4, 506-511.
[3] Manzocco L., Da Pieve S. & Maifreni M. 2011. Impact of UV-C light on safety and quality of fresh-
cut melon. Innovative Food Science and Emerging Technology, in press.

1732
Effect of ozonation on the sensory characteristics and pasting properties of cassava starch

Emanuele O. C. AMORIM, Vanessa C. DOVAL, Marcelo CRISTIANINI

Department of Food Technology (DTA), School of Food Engineering (FEA), University of Campinas
(UNICAMP), Campinas, SP, Brazil (olecram@fea.unicamp.br)

INTRODUCTION
The ozone is a strong antimicrobial agent with high reactivity and spontaneous decomposition
to non-toxic products [1]. Ozone has potential applications in the food industry and it can be
applied to food in liquid and gaseous forms [2]. Starch has been largely used in the food and
cosmetics industry due to its functional properties and its low cost. Being one of the most used
ingredients, starch imparts structure, texture, consistency and appeal to many food systems [3].
Previous works have shown that ozone is effective to decrease starch microbial contamination.
The objective of this study was to determine the effect of different ozone treatments on the
sensory characteristics and pasting properties of cassava starch.

MATERIALS & METHODS

Cassava starch (11% moisture) was humidified to obtain a final moisture content of
approximately 30%. The product was exposed to ozone in gas form (14 m3/h) pumped into a
horizontal dry mixer at approximate concentrations of 40 ppm for 30, 60, 90 and 120 minutes
and 118 ppm for 30 and 60 minutes. As a control, an experiment was carried out using only air
at the same flow rate. The samples were dried at 45°C for 24 h and their sensory characteristics
and pasting properties were evaluated.

RESULTS & DISCUSSION

On the pasting properties, the effect of ozone was more evident on the peak viscosity and
breakdown. The peak viscosity of the ozonized samples, with the exception of the sample
treated at 40 ppm for 30 minutes, was significantly higher than of the control and standard
samples. The ozonized samples presented lower cooking stability under agitation than the
untreated and control samples. The treatments that affected the greatest number of viscographic
characteristics were at 40 ppm/90 min. and 118 ppm/60 min. On the other hand, the sample
treated at 40 ppm/30 min. did not present any significant change on its pasting properties. The
difference between the two ozone concentrations employed was very discrete, and it was
noticeable only on the parameters breakdown, final viscosity and setback and basically in one
exposure time (60 minutes). The effect of increase in exposure time was observed only for the
parameters peak viscosity and breakdown, when the 40 ppm concentration was used, and for
the setback, when the 118 ppm concentration was employed.
Table 1 shows the mean notes for the color and odor attributes of the samples evaluated. The
results indicated that there was no significant difference among ozonized and standard samples
regarding the color, with the exception of the samples treated at 40 and 118 ppm during 30
minutes that were scored as higher (brighter) than the untreated sample.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1733
 Table 1. Mean values of the notes attributed to samples for the color and odor attributes
Samples
Attribute Exposure time
40 ppm 118 ppm Control Standard
30 minutes 1,42a 1,00a 1,42a 0,32b
60 minutes 0,64b 0,70 b
1,04a 0,39b
Cor
90 minutes 0,67b - 0,77a 0,40b
120 minutes 0,67a - 0,93a 0,57a
30 minutes 0,89a 1,07a 0,87a 0,39b
60 minutes 0,67a 0,70 a
0,54a 0,54a
Odor
90 minutes 0,95a - 0,37b 0,27b
120 minutes 0,99a - 0,47a 0,67a
In the same row, treated samples (ozonized and control) codified with different letters from the
standard sample differed significantly from this.

The effect of ozone on the odor attribute was more pronounced than on the color. The samples
treated at 40 ppm for 30 and 90 minutes and 118 ppm for 30 minutes were scored as different
from the untreated sample regarding the odor.

CONCLUSION
Previous work showed that ozone can be efficient to decrease starch microbial contamination.
However, the ozone treatment changed partially the pasting properties and sensory
characteristics of the product.

REFERENCES
[1] Kim J.G., Yousef A.E. & Dave S. 1999. Application of ozone for enhancing the microbiological
safety and quality of foods: a review. Journal of Food Protection, 62(9), 1071-1087.
[2] Dhillon B., Wiesenborn D., Wolf-Hall C. & Manthey F. 2009. Development and evaluation of an
ozonated water system for antimicrobial treatment of durum wheat. Journal of Food Science, 74(7),
396-403.
[3] Bhat R. & Karim A.A. 2009. Impact of radiation processing on starch. Comprehensive reviews in
food science and food safety, 8, 44-58.

1734
 Production of antioxidant enriched cranberry juice by electrodialysis with filtration

membrane: impact of process on juice composition

L. Bazinet1, S. Brianceau1, M. Araya-Farias1, Y. Desjardins2

1
Institute of Nutraceuticals and Functional Foods (INAF), Université Laval, Department of Food
Sciences and Nutrition, Laval University, Sainte-Foy (QC), Canada, G1V 0A6.
Laurent.Bazinet@fsaa.ulaval.ca
2
Institute of Nutraceuticals and Functional Foods (INAF), Université Laval, Department of Phytology,
Laval University, Sainte-Foy (QC), Canada, G1V 0A6

INTRODUCTION
Cranberry is recognized for its high antioxidant potential. So, the enrichment of a cranberry
juice with polyphenols, such as anthocyanins and proanthocyanidins, would be particularly
interesting for the ever growing nutrition-health market. To answer the consumer demands for
healthy products, the food industry is looking for innovative technologies for their
manufacture. Bazinet et al. [1] demonstrated that it is possible to enrich a cranberry juice in
antioxidants from another cranberry juice by electrodialysis with filtration membrane (EDFM).
This process is inexpensive, easy to use and environmently friendly. Nevertheless, in this first
preliminary work on the subject, it was demonstrated that if used as is this technology has the
disadvantage to generate a raw juice impoverished in antioxidants with an unsatisfactory color
and taste. In order to transpose this technology at an industrial scale for the production of an
antioxidant enriched cranberry juice, Bazinet et al.[1] proposed the integration of EDFM to the
conventional process used for cranberry juice production, in a way avoiding the generation of a
source juice impoverished in polyphenols. However, the effectiveness of such a process was
not demonstrated.
In this context, the objectives of this work were 1) to validate the feasibility of the integrated
proposed process, 2) to study the composition evolution of source and enriched cranberry
juices during consecutive EDFM treatments 3) to study the EDFM parameters evolution.

MATERIALS & METHODS

EDFM treatments were performed in batch process using a constant voltage difference of 30 V.
In order to evaluate the feasibility of EDFM treatments for the production of a polyphenol-
enriched cranberry juice 300 mL of cranberry juice were circulated in the compartment on the
cathode side and 3L of cranberry juice in the compartment on the anode side. The cranberry
juices were circulated three times (3 cycles). The duration of the treatment was 2 h.
Conductivity and pH of cranberry juices were recorded throughout the process as well as the
global system resistance. Anthocyanin, proanthocyanidin, organic acid, mineral contents (Na,
K, P, Ca, Mg, Cu) juice color parameters (L*, a* and b*) and °Brix were determined before
treatments and following each cycle on the cranberry juices and on a control.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1735
RESULTS & DISCUSSION
The general trends in the different juices were that the anthocyanin contents increased in the
enriched juice all along the treatment, while at the same time the concentration of anthocyanins
were constant whatever the cycle. Total anthocyanin concentration in the enriched juice
increased from 136.4 ± 7.1 to 162.9 ± 8.0 g/L during treatments corresponding to a 19.4%
enrichment value. However, total concentration in the raw juice remained constant at 125.9 ±
6.3 g/L throughout the treatment. Organic acid concentrations remained constant and were
preserved in treated cranberry juices except for citric acid concentration. EDFM treatments did
not have a significant effect on the pH of the enriched and raw cranberry juices. The mean pH
of enriched and raw juices was 2.6 ± 0.1 and 2.9 ± 0.1 respectively. Soluble sugars (Brix) in
juices remained constant at 6.9 ± 0.5 throughout treatments. Color parameters for the enriched
juices remained constant during all cycles of EDFM. The mean global system resistance
increased from 50.3 ±1.5 Ÿ to 83.0 ± 4.0 Ÿ during the first cycle and from 65.3 ± 2.4 Ÿ to 87.3
± 10.3 Ÿ during the second and third cycle. The decrease in the global system resistance at the
beginning of each cycle might also be due to a fouling of the membranes. During each cycle,
the decrease in the electrical conductivity of ion-exchange membranes resulted in an increase
of the system resistance.

CONCLUSION
Results obtained in this study showed the effectiveness of the process to increase the
antioxidant capacity and polyphenol content of a cranberry juice and confirmed the possibility
to use EDFM process for the production of an antioxidant-enriched cranberry juice fraction
from a large juice source. The model proposed for the integration of an EDFM system to the
process used for the production of cranberry juices can be transposed at an industrial scale.
However, prior to the development of an industrial process, it will be essential to carry-out a
study on the fouling of the membranes and determine the evolution of membrane parameters
such as the streaming potential determination and the identification of the molecules interacting
with the different types of membranes.

REFERENCES
[1] Bazinet L., Cossec C., Gaudreau H. and Desjardins Y. (2009). Production of a phenolic antioxydant
enriched cranberry juice by electrodialysis with filtration membrane, J. Agric. Food Chem. 57(21) :
10245-10251.

1736
 Effect of sunflower oil applied by vacuum impregnation to refrigerated atlantic salmon

Puente, L.a, Ortiz, J.a, Leiva, M.a, Aubourg, S.b

a
Universidad de Chile, Food Science And Chemical Technology, Santiago, Chile (lpuente@ciq.uchile.cl)
b
IIM-CSIC, Biotechnology and Acuiculture, Vigo, España (saubourg@iim.csic.es)

INTRODUCTION
Salmon can be considered as a natural functional food, highly consumed due to their pink meat
and a high content of poliinsaturated fatty acids omega-3 (acid eicosapentaenoic (C20:5, EPA)
and acid docosahexaenoic (C22:6, DHA). These functional properties can be increased by
adding some antioxidant compounds to improve their stability against oxidative deterioration.
One of the novel technologies to obtain new functional foods are vacuum impregnation,
technique based on the addition of physiologically active compounds to food matrix by means
of the hydrodynamic mechanism.

MATERIALS & METHODS

In this work we used the vacuum impregnation in two different matrices, fillets and gels of
Atlantic salmon, we studied the effect using two impregnating solutions, sunflower oil as a
source of alpha-tocopherol and a mixture of this oil with rosemary extract . The effects on the
development of oxidative rancidity (peroxide value and p-anisidine value) and the content of
alpha-tocopherol were measured in both arrays in cooling additionally measured astaxanthin
content and microbiological growth (RAM).

RESULTS & DISCUSSION

The effects on the development of oxidative rancidity (peroxide value and p-anisidine value)
and the content of alpha-tocopherol were measured in both arrays in cooling additionally
measured astaxanthin content and microbiological growth (RAM) . The results indicated
differences in the penetration of substrate and the protective effect of rancidity during storage
depending on the type of matrices, gel fillet salmon lipoperoxidation parameters indicated
higher levels of primary and secondary oxidation in the gel, possibly associated the
characteristics of the vehicle through impregnation and tocopherols, also existed but
astaxanthin degradation caused by the impregnation process of the fillets and the effect of the
gelation temperature, affecting the color. On the other hand, the impregnation of sunflower oil
and rosemary extract showed some antimicrobial activity

11th International Congress on Engineering and Food - Athens, Greece, 2011 1737
a) b)

meq Oxígeno/Kg
6 5
meq oxígeno/Kg

5 4
4 3
3 2
2 1
1 0
0 0 5 10 15 20 25 30
0 5 10 15 20 25 30 Tiempo [días]
Tiempo [días]

Impregnación aceite Impregnación aceite + extracto Romero Control Impregnacióna aceite impregnación aceite + extracto Romero Control

Figure 1. Peroxide value for salmon fillets and gel.

CONCLUSION
After the results we can conclude that the impregnation of sunflower oil and rosemary extract
may reduce rancidity and inhibit microbial growth in chilled Atlantic salmon.

1738
 Production of Mucor griceocyanus protease using different carbon sources in submerged

fermentation

a
Alejandro Ramírez, aJose Sánchez , aAnna Iliná, bJulio C. Dusted Mendoza, aJ. Rodríguez, aJosé L.
Martínez

a
Dpto. de Biotecnología, Facultad de Ciencias Químicas. Universidad Autónoma de Coahuila. Saltillo,
Coahuila, México. bGrupo de Biotecnologia, Facultad de Ingeniería Química, Instituto Superior
Politécnico José A. Echeverría. Habana Cuba. (jose-martinez@uadec.edu.mx)

INTRODUCTION
At present, the enzymes are the bigger market covering 60% of this industry are proteases,
enzymes that break peptide bonds [1,3,4]. These enzymes are found naturally in living
organisms, plants, animals and microorganisms (fungi, bacteria and yeast). The applications of
proteases are various as production of cheese and elaboration of detergents. The biotechnology
and bioengineering have enabled the collection and production of proteases of microbial origin.
Some Mucorales species have been reported as protease producers but few papers have been
published about the variables that control the production of protease from Mucor species [1,3].
In this study we evaluated the ability to synthesize Mucor griseocyanus proteases in submerged
culture using different carbon sources (glucose, lactose and whey).

MATERIALS & METHODS

Enzyme activities were compared by testing the synthetic media glucose and lactose
concentrations of 40, 60 and 80 g/L, in the case of whey was tested without modification. It
also assessed the effect of initial pH over the protease production. Finally, we carried out a
fermentation bioreactor scale (4L) under optimal conditions [2]. The optimal pH and
temperature of protease of Mucor griseocyanus was evaluated.

RESULTS & DISCUSSION

This study allowed to demostrate the ability of M. griseocyanus to produce extracellular
protease enzyme in submerged fermentation using different carbon sources at different
concentrations.

Figure 1. Maximum levels of protease activity recorded using glucose, lactose and whey as carbon
source.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1739
According to Figure 1, in glucose medium at concentration of 40 g/L the protease activity level
in tyrosine units (TU) was 29574 U/L at 24 h. Subsequently presented a second peak of 20090
UT / L at 48 h. A similar profile was observed when using lactose, showing a level of
maximum protease activity 16200 U/ L at 24 h, and 25038 U/L at 48 h, being the highest level
of activity registered. This behavior suggests the presence of isoenzymes or different types of
protease that is synthesized at different times of growth. At concentration of 60 and 80 g / L is
observed the same profile, however, low protease activity was detected (Fig. 1). Is possible
catabolic repression.

It was determined that the best medium for protease production with small-scale Mucor
griseocyanus was whey, as it presented a volumetric productivity of 648 U/L * h. The best
conditions of pH and temperature for protease activity of M. griseocyanus were: 4-5 and 30 °C,
respectively. The maximum level of protease activity produced by M. griseocyanus the 48 h of
fermentation in a 4 L fermentor (Fig. 2), by using whey as the sole source of nutrients enriched
with trace salts, was 27,400 U/L.

Figure 2. Protease production by M. griseocyanus using whey as a source of nutrients in 4-liter reactor.

CONCLUSION
It was found that M. griseocyanus synthesized proteases when used in submerged fermentation
using as carbon sources: glucose, lactose and whey. Concentrations evaluated permit highest
levels of enzyme activity using a carbohydrate concentration of 40 g / L, however, levels in the
whey were slightly higher. The whey permit to obtaining higher levels of enzyme protease,
therefore, favor the development of economic growth media, because it is a byproduct of the
dairy industry and generates high costs in its treatment to reduce environmental impacts.

REFERENCES
[1] Alves H. M., de Campos-Takaki M. G., Okada K., Ferreira H. I. and Milanez A. I. 2005. Detection of
extracelular protease in Mucor species. Rev. Iberoam. Micol., 22: 114-117.
[2] Coca, J. & Dustet J. 2006. Expression and characterization of lipase produced by Mucor griseocyanus.
La Habana, Cuba. En: Biotecnología Aplicada, 23:224-228.
[3] Haq, I., Mukhtar H. and N. Riaz, 2004. Protease biosynthesis by mutant strain of Penicillium
griseoroseum and cheese formation. Pakistan J. Biol. Sci., 7: 1473 – 6.
[4 ] Haq, I., Mukhtar H., Umber H. 2005. Production of Protease by Penicillium chrysogenum Through
Optimization of Environmental Conditions. Journal of agriculture & social sciences, 2(1): 23-25.

1740
 Evaluation of MAP design parameters on quality of fresh-cut produce

F. Oliveira12, M. Sousa-Gallagher1, P. Mahajan1, J. Teixeira2

1
Department of Process & Chemical Engineering, University College Cork, Ireland
(feliz.oliveira@hotmail.com, m.desousagallagher@ucc.ie, p.majahan@ucc.ie)
2
IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of
Minho, Portugal (jateixeira@deb.uminho.pt)

INTRODUCTION
MAP design depends of the characteristics of the product, its recommended gas composition
and its respiration rate as affected by temperature and headspace gas composition; and the
permeability of the packaging materials (perforated or non-perforated polymeric film) and its
dependence on temperature. An inappropriately designed MAP system may be ineffective or
even shorten the storage life of a product: if the desired atmosphere is not established rapidly,
the package draws no benefit; if O2 and O2/CO2 levels are not within the recommended range,
the product may experience serious alterations and its storage life may be shortened. The
objectives of this work were to determine the effect of the MAP design parameters (number of
perforations, weight of CO2 scavenger and amount of product) on fresh sliced mushrooms,
based on headspace gas composition and quality parameters changes; and to determine number
of perforations, weight of CO2 scavenger and amount of mushrooms to be packed, to obtain
optimum MAP conditions for increasing the shelf life of the product.

RESULTS AND DISCUSSION

Since mushrooms have high respiration rate, equilibrium modified atmosphere was achieved
within one day. The number of perforations, amount of CO2 scavenger and weight of sliced
mushrooms showed a significant effect on headspace gas composition, as shown in Figure 1a.
Pareto chart shows that the presence of CO2 scavenger had no significant effect on package O2
concentration; on the contrary, it had a significant effect on package CO2 concentration (Figure
1b).
All factors studied had a significant effect (p < 0.05) on weight loss of mushrooms. Number of
perforations and amount of CO2 scavenger had a significant effect (p < 0.05) on pH, whereas,
the weight of mushrooms within the package did not have a significant effect on pH. The
number of perforations had a significant effect on firmness and colour of mushrooms.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1741
 Figure 1. a) Surface plots with effects of number of perforations (N),weight of CO2 scavenger (S) and amount of
mushrooms (W) on 1) headspace O2 composition and 2) headspace CO2 composition, after 3 days at 10 ºC. b) Pareto
charts of standardized effects for number of perforations, weight of CO2 scavenger and amount of product on 1)
headspace O2 composition and 2) headspace CO2 composition, at 95 % significance level, indicated as a vertical
dashed line.

Figure 2. Quality parameters of sliced mushrooms at time 0 and after 3 days in the package with 3 perforations in
the film, 1 g of CO2 scavenger and 100 g of sliced mushrooms.

CONCLUSIONS
Optimisation of MAP design interplays for the best headspace gas composition and
consequently lower changes in quality parameters of the product. This work showed that at
lower O2 in the headspace composition, obtained in packages with 1 perforation, the quality
changes were smaller. Adding CO2 scavenger in the package increased the deterioration of
mushrooms. Good MAP design requires consideration of weight of mushrooms to be packed
and number of perforations in the film (i.e., 110 g of sliced mushrooms, 2 perforations) in order
to achieve the best headspace gas composition (3.6 % of O2 and 11.5 % of CO2, after 3 days at
10 ºC) and to extend the shelf life of fresh sliced mushrooms.

1742
 Rational method for designing efficient food separation processes by chromatography.

“Polyphenol-ethanol/water system with polymer-resins”

Mareto Hosono, Ryo Maeda, Noriko Yoshimoto, Shuichi Yamamoto*

Bio-Process Engineering Laboratory, School of Engineering & Graduate School of Medicine
Yamaguchi University, Ube 755-8611, Japan (shuichi@yamaguchi-u.ac.jp)

INTRODUCTION
A method for predicting efficient conditions for polyphenol separations by
polymer-resin-based chromatography. Our concept “iso-resolution curve”
originally
developed for linear gradient elution (LGE) chromatography of proteins[1], which is useful for
understanding the relationship between separation time and solvent consumption was extended
to stepwise (isocratic) elution chromatography. For the iso-resolution curve calculations the
distribution coefficient K as a function of ethanol concentration I was determined by using our
LGE model[1]. The HETP was measured as a function of mobile phase velocity, u for different
I. From the HETP-u curves the pore diffusivity was determined as a function of I. The
iso-resolution curves calculated with these parameters were examined in order to understand
the separation process and optimize the process.

MATERIALS & METHODS

The experimental setups and protocols were essentially the same as in our previous study[2].
Polystyrene-divinyl benzene (PS-DVB) resin particles (Diaion HP20SS, Mitsubishi Chemicals)
were packed into a column (diameter dc=1.1 cm, packed bed height Z = 15-20 cm). The particle
diameter dp was measured microscopically as a function of I; dp = 62 Pm at I =0.2, dp =
67 Pm at I=0.3 and dp = 72Pm at I=0.4. The model polyphenol samples were catechin
(C15H14O6 mol.wt. 290) and epigallocatechin gallate (C22H18O11, mol.wt. 458) abbreviated as
EGCG. The mobile phase was an ethanol-water mixture and the ethanol concentration is given
by I [ethanol volume/(ethanol volume + water volume)]. The temperature was maintained at
298r2K.
Isocratic elution experiments were carried out as a function of u, and the peak retention time tR
(or retention volume VR ) and the peak width (standard deviation) Vwere measured. Then,
HETP and K were calculated according to the following equations.
HETP = Z(V/tR)2 (1) VR=FvtR=Vo+(Vt-Vo)K = Vo(1+HK) (2)
Fv=volumetric flow-rate, Vo = void volume, Vt = bed volume. H=(Vt-Vo)/Vo=(1-H)/H is the
volumetric phase ratio. u is given by Fv/(AcHwhere Ac=column bed cross sectional area.

RESULTS & DISCUSSION

Figure 1 shows typical elution curves of catechin and EGCG. Although decreasing I resulted in
better separations, the peak became wider and the elution volume increased. The elution curve
became also wider with increasing u while the peak retention volume remained constant. HETP
was expressed as a function of u by the following equation.
HETP = Ao +Cou = Ao + HKdp2u/[30DS(1+HK)2] = Ao + HK2dp2u/[30DDm(1+HK)2] (3)
o
A is the axial dispersion term (=0.04 cm). The pore diffusion coefficient DS was found to be

11th International Congress on Engineering and Food - Athens, Greece, 2011 1743
expressed by the molecular diffusion coefficient
Dm and K as DS = DDm/K = 0.07 Dm/K (4)
The iso-resolution curve for a given column
length Z can be calculated by Eq.(5).
Rs=(tR2-tR1)/[0.5(W1+W2)] where W is the baseline
peak width W=4V
Z H ( K 2  K1 )
Rs 1
2 (1  HK ) A o  C o u  (1  HK ) A o  C o u
1 1 1 2 2 2
(5)
The subscript 1 and 2 imply the component 1 and
2 respectively (The component 1 elutes first).
The separation volume (solvent consumption)
VSEP and the separation time tSEP are given by
V SEP V0 (1  HK 2 )  ( FV W 2 ) / 2
V0 (1  HK 2 )  2V0 (1  HK 2 ) ( A2 o  C 2 o u ) / Z (6)

t SEP V SEP / Fv (7)

The calculated iso-resolution curves for different

column lengths are shown in Fig.2. It is shown
that there is a region where both VSEP and tSEP are
small. In order to verify the iso-resolution curve,
experimental elution curves are compared with the
calculated ones (Fig.3). The agreement between
the experimental and calculated curves is
reasonable when we consider the errors involved
in the correlation equations for K and HETP. Our
proposed method is easy to use and allows to
design an economically feasible food separation
process by polymer resin adsorption
chromatography.

CONCLUSION
A method for calculating the iso-resolution curve
was developed and verified experimentally for the
polyphenol-ethanol/water system with poly
styrene divinylbenzene (PS-DVB) resin
chromatography. The iso-resolution curve was
found to be useful for determining suitable
(economically feasible) separation conditions.

REFERENCE
[1] Yamamoto S. & Kita A. 2005. Theoretical background of short chromatographic layers: Optimization of gradient
elution in short columns. Journal of Chromatography A, 1065, 45-50. [2] Yamamoto S., Hakoda M., Oda T. & Hosono
M. 2007. Rational method for designing efficient separations by chromato- graphy on polystyrene-divinylbenzene
resins eluted with aqueous ethanol. Journal of Chromatography A, 1162, 50-55

1744
 Food-grade emulsions prepared by membrane emulsification techniques

F. Spyropoulos, R.D. Hancocks, I.T. Norton

School of Chemical Engineering, University of Birmingham, UK ( F.Spyropoulos@bham.ac.uk)

INTRODUCTION
Membrane emulsification is a relatively new technique which attempts to improve on
traditional emulsification methods by producing each droplet singly, as required (1, 2). The aim
of this study was to explore the full potential of membrane emulsification in the production of
food emulsions. The effect of both the process and the material parameters on the structure of
oil-in-water (o/w) emulsions was investigated.

MATERIALS & METHODS

In all produced oil-in-water (o/w) emulsions, the oil phase was commercially available
sunflower oil and the water phase was passed through a reverse osmosis unit and a milli-Q
water system. Tween 20 and 80, used to stabilise the emulsions, were all purchased from
Sigma Aldrich (Dorset, UK). The SPG and ceramic membranes used were purchased from
SPG Technologies (Japan) and TAMI industries (France), respectively. Droplet sizes (D4,3) of
all emulsions were measured using a Malvern Mastersizer (UK).

RESULTS & DISCUSSION

Effect of trans-membrane pressure
The data obtained show that increasing the trans-membrane pressure (¨PTM) results in an
increase in the droplet size of produced emulsions but also their droplet size distribution. ¨PTM
must be above a critical pressure before any droplets are produced. At low ¨PTM (just above
critical) the smallest droplets are produced, but the flux is generally very low. Increasing the
¨PTM increases flux, but also droplet size and size distribution. The observed behaviour was
found to be membrane type dependant.
Effect of cross-flow velocity
In general, increasing the cross-flow velocity results in “earlier” droplet detachment and thus
smaller droplet size. The size of droplets prepared with SPG membranes appear to be more
“responsive” to changes in cross-flow velocity. At higher cross-flow velocities the mean
droplet size/mean pore size ratio is less than 2:1, compared to the generally achievable ratio of
around 5:1(3).
Effect of emulsifier type and concentration
Increasing the concentration of Tween 20 and 80 emulsifiers has a similar effect on the o/w
droplet size reduction (Table 1). In both cases, an equilibrium droplet size is produced with
about 0.2% emulsifier.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1745
Effect of dispersed phase volume fraction
The effect of dispersed phase volume fraction (up to 70%) on the droplet sizes of emulsions
stabilised using varying concentrations of Tween 20, was investigated. It was shown that for
volume fractions of up to 50% there was only a small increase in droplet size with further
volume fraction increases giving higher droplet sizes. The latter is suggested to be due the
strong tendency of the system, at such high dispersed phase volume fractions, to phase invert.
Comparison between membrane emulsification and high shear mixing
Equilibrium droplet size using membrane emulsification is achieved for a 0.2% emulsifier
rather than about a 1% when processing using a high shear mixer (Table 1). This suggests the
membrane emulsification is more “efficient” in its use of emulsifier, i.e. the same droplet size
can be achieved using less emulsifier than what is required in conventional processes. This is
likely due to the lower rate of interfacial area generation induced when processing by
membrane emulsification in comparison to the more “explosive” rate of interfacial area
generation associated with systems processed by droplet break-up equipment.

Table 1. Average droplet size of emulsions produced with different emulsifiers using either membrane
emulsification or high shear mixer (Silverson).
Membrane Emulsification High shear mixer
Tween 20 Tween 80 Tween 20

Emulsifier concentration [wt. %] D4,3 [Pm] D4,3 [Pm] D4,3 [Pm]

0.01 26.12 r 6.41 26.51 r 7.02 29.08 r 8.35
0.1 11.81 r 3.24 12.91 r 3.85 20.34 r 4.29
0.2 7.44 r 2.11 8.14 r 2.25 14.46 r 3.12
0.6 6.26 r 1.88 6.46 r 1.85 8.73 r 2.34
1 5.91 r 1.85 6.22 r 1.83 7.42 r 2.28

CONCLUSION
Membrane emulsification can produce food emulsions with a wide range of droplet sizes,
droplet size distributions and dispersed phase volume fractions. Both process (e.g. cross-flow
velocity) and material (e.g. emulsifier concentration) parameters can be carefully chosen to
control the emulsion microstructure produced. Several advantages of the studied process over
conventional comminution techniques were demonstrated including the ability to produce
stable emulsions with lower (up to 80%) emulsifier concentration.

REFERENCES
[1] Charcosset C., Limayem I. & Fessi H. 2004. The membrane emulsification process - a review. Journal
of Chemical Technology & Biotechnology, 79(3), 209-218.
[2] Joscelyne S.M. & Trägårdh G. 2000. Membrane emulsification - a literature review. Journal of
Membrane Science, 169(1), 107-117.
[3] Schadler V. & Windhab E.J. 2006. Continuous membrane emulsification by using a membrane system
with controlled pore distance. Desalination, 189(1-3), 130-135.

1746
 Use of supercritical CO2 for the inactivation of Aspergillus niger inoculated on stainless

steel plates surface

Mariana Altenhofen da Silvaa, Juliana de Souza Ferreirab, Beatriz Thie Iamanakac, Fabiana Sayuri
Kiharaa, Ricardo Soares Cutoloa, Theo Guenter Kieckbuscha

a
School of Chemical Engineering, University of Campinas, Campinas, Brazil (theo@feq.unicamp.br)
b
Department of Chemical Engineering, Federal University of Uberlandia, Uberlandia, Brazil
c
Laboratory of Microbiology, Institute of Food Technology (ITAL), Campinas, Brazil

INTRODUCTION
The use of supercritical carbon dioxide (SC-CO2) for the inactivation of microorganisms
continues to attract attention as an alternative non-thermal technique for pasteurization and
sterilization of food products and packaging materials. SC-CO2 is expected to induce only
minimal thermal degradation and change in product quality because the process is performed
under relatively low pressure and temperature [1].
In high pressure carbon dioxide (HPCD) processing, the products are contacted with either sub-
or supercritical CO2 for a designated time in a batch, semi-batch or continuous manner.
Supercritical CO2 is CO2 at a temperature and pressure above its critical point values (Tc
=31.1°C, Pc =7.38 MPa), while for subcritical (gaseous or liquid) CO2 the temperature or
pressure are below its thermodynamic critical point values [2].
Using CO2 as a sterilizing agent has several potential benefits. Carbon dioxide is a non-toxic
and inexpensive gas and has been considered in the sterilization of a variety of heat sensitive
products[3].
In the present study, preliminary tests to investigate the sterilizing potential of SC-CO2
treatment against Aspergillus niger inoculated on the surface of stainless steel plates were
conducted. This was done with the purpose to adapt the experimental apparatus and validate
the methodology for SC-CO2 treatment, as an alternative sterilizing method, mainly for
materials that need less aggressive treatments.

MATERIALS & METHODS

The use of high pressure CO2 as a microbial inactivating agent less aggressive to a widespread
of goods is considered. A semi-continuous flow apparatus, operated in a batch mode with
independent control of temperature and pressure was used in the inactivation experiments.
Several operational changes were necessary in order to adapt the SC-CO2 experimental set-up
for sterilization process. Stainless steel plates (1 cm x 1cm) inoculated with Aspergillus niger
(» 105 CFU/mL) were transferred to a sterile support and carefully put inside the sterilization
chamber. At the end of each experiment the pressure vessel was opened under aseptic
conditions and the metal plates were collected and submitted to viable surviving spores
counting by the standard plating technique on DG18 agar plates. Results of SC-CO2 treatments
to attain sterilization of Aspergillus niger at a temperature of 30ºC, one pressure cycle up to 10,
7.5 and 6 MPa without retention time using an adapted pressure vessel are reported.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1747
RESULTS & DISCUSSION

(a) (b)

Figure 1. Experimental set-up of the SC-CO2 sterilization system before (a) and after (b) the operational
changes.

Table 1. Efficiency of the CO2 treatment (6, 7.5 and 10 MPa/30°C/time for system pressurization and
depressurization) in the inactivation of Aspergillus niger inoculated on stainless steel plates surface
Count before Count after treatment
Pressure Treatment time (min)
treatment (CFU/mL)
(MPa)
Pressurization Depressurization (CFU/mL) After test After 4 weeks
5
10.0 15 5 2.13·10 ND* ND
5
7.5 6 2 2.68·10 ND ND
3 2
6.0 0.5 0.3 7.50·10 2.25·10 NC**

*ND: not detected; **NC: not concluded

CONCLUSIONS
Results confirm the ability of SC-CO2 to inactivate Aspergillus niger inoculated on metal
plates’ surface under mild conditions (10 and 7.5 MPa and 30°C). A reduction of 1.5 log cycles
was attained using an experimental pressure of 6 MPa (subcritical condition for CO2). Further
investigations are under way in order to evaluate the effect of subcritical CO2 pressures with
different treatment times in the inactivation of Aspergillus niger.

REFERENCES
[1] Zhang J., Davis T.A., Matthews M.A., Drews M., Laberge M. & An Y.H. 2006. Sterilizing using
high-pressure carbon dioxide. The Journal of Supercritical Fluids, 38, 354-372.
[2] Garcia-Gonzalez L., Geeraerd A.H., Elst K., Van Ginneken L., Van Impe J.F. & Devlieghere F. 2009.
Influence of type of microorganism, food ingredients and food properties on high-pressure carbon
dioxide inactivation of microorganisms. International Journal of Food Microbiology, 129, 253–263.
[3] Ballestra P. & Cuq J. 1998. Influence of pressurized carbon dioxide on the thermal inactivation of
bacterial and fungal spores. Lebensmittel Wissenschaft und Technologie, 31 (1), 84-88.

1748
 Non-aqueous thermal processing of foods

Robert Steele a and Clemence Kerjeana

a
CSIRO Food and Nutritional Sciences, North Ryde, Australia (Bob.Steele@csiro.au)

INTRODUCTION
Water has been conventionally used as the heat transfer medium for both heating and cooling
because of its excellent heat transfer properties, low cost and easy disposal. Its continuing use
should be questioned because of the predicted shortage of water in some countries. The
purpose of this investigation was to examine the potential of non-aqueous thermal processing
to deliver safe, wholesome food. No commercially available air-based retorts were available
for this study so measurements were made at T< 75°C
The following goals were addressed:
x To compare fan-forced air and water as heating media.
x Does non-aqueous thermal processing affect the properties of the plastic containers?
x Is the quality of food more adversely affected by heating in hot air than in water?
Plastic packaging often includes a barrier layer of ethylene vinyl alcohol copolymer film
between polypropylene layers. The barrier properties of this layer are known to be
compromised by the inevitable hydration that accompanies aqueous thermal processing.
Non-aqueous thermal processing offers an environmentally friendlier way to process foods
which may reduce costs and may prevent compromising the barrier.
Most attempts at developing non-aqueous thermal processing were aimed at developing
processes for improving the quality of the food. Concerns with respect to the extensive use and
cost of process water were not the serious environmental issue they are today.

MATERIALS & METHODS

Three container structures were studied:
Pack 1. A rectangular parallelepiped aluminium pack with dimensions 73.5 X 73.5 X 25 mm,
average thickness = 0.13 mm.
Pack 2. A rectangular parallelepiped plastic pack 64.4 X 64.4 X 23.4 mm, average thickness
0.44 mm, with PP/NY MXD6 /PP layers.
Pack 3. A plastic truncated cone h = 47.6 mm diam = 69.2 mm, average thickness 0.50 mm
with PP/EVOH/PP layers.
The packs were filled with cream style corn. Temperatures were measured at six points of
each of the containers, three in the middle and three near the inner wall.
Aqueous processing
Packages were immersed in a water bath with a metal grid to ensure full immersion.
Non-aqueous processing
Packages were placed in a fan-forced hot air oven and centre temperatures measured at two
velocities : 0.2m.s-1 and 1.6m.s-1.
Cooling was not studied in this project.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1749
Oxygen rate transmission (OTR) was determined using a Mocon (OX-TRAN® Model 2/21
oxygen permeability) by ASTM F1307 at 21°C after two hours and repeated at approximately
24 h intervals for 5 days. In this study the OTR is reported as ml 02.day-1.pkg-1.atm-1 .
Product quality the colour change of the corn was measured using a colorimeter (Minolta
chroma meter CR-400/410) in triplicate before and after heating

RESULTS & DISCUSSION

Air heating took longer than water for the unaccomplished temperature differential to reach
within 0.01 K of the heating medium (Tm) consistent with the findings of Ekelund et al1 .
Increasing the velocity of heated air from 0.2 to 1.5 m.s-1decreased the fh of pack 3 from 100
to 50 min. Plastic containers, with a low thermal conductivity (0.45 W.m-1.K-1) and are
comparatively thick, approach Tm more slowly than the aluminium containers, irrespective of
which heating medium is used. Despite aluminium’s high thermal conductivity (237 W.m-1.K-
1
) the advantage of using aluminium is diminished when using a low heat transfer fluid such as
air. The fh of the plastic packs was about 16% greater than aluminium packs when heated in
water however in air shows that the thermal advantage of using aluminium was slightly
diminished to 13%.The colour change was more marked for air-heated plastic packs than for
water-heated packs.
After heating, water-heated packs had about twenty times the OTR of the untreated packs.
While the 0TR decreased with the time it did not return to its initial value, consistent with the
findings of Tsai & Jenkins (1988). On the other hand after air heating, the OTR remained low
~ 0.02 as shown in Figure 1.

Figure 1. OTR of plastic packs heated at 60 C in air and water

CONCLUSION
Hot air is a cheap, simple and safe heating medium than has some advantages over water or
steam especially the retention of barrier properties of EVOH barriers. The expected
environmental gain may allow this process to replace conventional steam and water retorts
however further studies at temperatures >100ºC are needed.

REFERENCES
[1] Ekelund, E.A., Frisk, P., and Berglund, S.D.T..1961. New methods of sterilizing heat on hot air
sterilization and heat transfer. Int. Congr. Cann. Fds 4th, 125.

1750
 Estimation of Peroxidase Activity in red cabbage by Artificial Neural Network (ANN)

Iman Shahabi Ghahfarrokhia, Amir Daraei Garmakhany b, Seied Mohamad Mousavi c

a
Islamic Azad University- Shahrekord Branch, Food Science and Engineering Dept, Shahrekord, Iran
(shahabi_62@yahoo.com )
b
Gorgan University of Agricultural Sciences and Natural Resources, Food Science and Engineering
Dept, Gorgan, Iran (amirdaraey@yahoo.com )
c
University of Tehran, Food Science and Engineering Dept, Tehran, Iran (mousavi@ut.ac.ir)

INTRODUCTION
Peroxidase (POD) is an enzyme commonly found in vegetables which cause off-flavors and
colors in raw and un-blanched frozen vegetables [1]. Inhibition of the enzyme activity in fruits
and vegetables is generally achieved using physical or chemical treatments such as heating
(blanching), lowering pH and/or aw or adding chemical additives. Due to the consumer market
demands which is concerned about the use of chemicals in such products, more attention has
been given to the search for alternative anti browning compounds [2]. Mathematical model is
sometimes difficult to realize since created models, based on microbial or enzyme kinetics, are
very often highly complex. Artificial Neural network (ANN) are universal approximators
which require relatively little time to construct and do not require any prior knowledge [3].

MATERIALS & METHODS

Ten grams of each vegetable and water (30ml) were homogenized for 3 min. The slurry was
filtered and centrifuged. The supernatant was used as the enzyme source. Peroxidase activity
was determined spectrophotometrically at 470nm using guaiacol and H2O2 [2] One unit of
activity is defined as a change in absorbance of 0.001 min-1. In total, 240 data were collected
for the three different essential oils (Cumin, Fennel and Clove), four antioxidant concentrations
(50, 75, 100 and 200 Pl / 100ml ) and 20 durations of enzyme activation (0, 20, 40… 400
seconds) as input neuron and enzyme activity as output neuron. the data order was randomized
and divided to training data (40% of data), crossvalidation data (30% of data), testing data
(30% of data) for. During training, momentum value was fixed at 0.7, and learning rate was
determined at level 1 on the hidden layer and 0.1 at the output layer. The training process was
carried on for 65000 epochs or until the cross-validation data’s MSE, did not improve for 100
epochs to avoid over-fitting of the network.

RESULTS & DISCUSSION

The ANN parameters used for prediction of POD activity in red cabbage are shown in Table 1.
The optimal number of hidden layers and number of neurons in the hidden layers were selected
by using a trial and error method based on minimizing the difference between estimated ANN

11th International Congress on Engineering and Food - Athens, Greece, 2011 1751
outputs and experimental values. It was found that ANN with one hidden and 21 hidden
neurons had the minimum MSE for training and cross validation 7.9815 u 10 6
and 2.9123 u 10 5 , respectively. POD activity was shown the highest sensitivity to time. It is
clear; all chemical reaction is dependent to time. Between 3 essential oils, cumin was shown
the highest effect on POD activity due to high antioxidant properties

Table 1. The best structure and optimum values of the ANN produced in testing stage

Step size Momentum Transfer function Testing

ANN ANN
Hidden Output Hidden Output Hidden Output Epoch
structure model` MSE NMSE MAE R2
layer layer layer layer layer layer
Linear
3-21-1 MLP 1 0.1 0.7 0.7 Tanh 65,000 0.0002629 0.0026579 0.0102657 0.9974
Sigmoid

CONCLUSION
The POD activity of vegetable and fruits is a linear phenomenon which depends on several
factors. ANNs are mathematical models whose architecture has been inspired by biological
neural networks. ANNs are very appropriate for the modelling of linear and non-linear
processes. The advantage of ANNs over conventional method like regression test is time and
cost saving. Also, the ANNs can consider more input parameters and the performance is better
than conventional methods. The result of this study showed that The ANN model predicted
POD activity with MSE 0.0002629 and good correlation between predicted and experimental
data (R2= 0.9974). These results show the ability of ANN technology for predicting POD
activity of red cabbage under natural antioxidants. Results showed that POD activity was the
highest sensitivity to time and between 3 essential oils, cumin was shown the highest effect on
POD activity due to high antioxidant properties

REFERENCES
[1] Lee H.C. & Klein B.P. 1989. Evaluation of combined effects of heat treatment and antioxidant on
peroxidase activity of crude extract of green peas. Food Chemistry 32, 151-158.
[2] Daraei Garmakhany A., Mirzaei H. O., Aghajani N. & Kashiri, M. 2010. Investigation of natural
essential oil antioxidant activity on peroxidase enzyme in selected vegetables. J. Agri Scie and
Techno USA., 4, 78-82.
[3] Bryjak J., Ciesielski K. & Zbicinski I. 2004. Modelling of glucoamylase thermal inactivation in the
presence of starch by artificial neural network. Journal of biotechnology 114, 177-185.

1752
 Quality classification of corn tortillas by means of cross validation between sensorial
evaluation and computer vision system

José Jorge Chanona-Péreza, Domingo Meryb, Alvaro Sotob, José Miguel Aguilerab, Aldo Ciprianob,
Nayeli Veléz-Riveraa, Israel Arzate-Vázqueza, Gustavo Fidel Gutiérrez-Lópeza

a
Departamento de Graduados e Investigación en Alimentos, Escuela Nacional de Ciencias Biológicas,
Instituto Politécnico Nacional, Carpio y Plan de Ayala s/n, Santo Tomás 11340,
D.F. México, México (jorge_chanona@hotmail.com).
b
Escuela de Ingenieria, Pontificia Universidad Catolica de Chile, Av. Vicuna Mackenna 4860 (143),
Macul, Santiago de Chile, Chile.

INTRODUCTION
In Mexico, tortillas are consumed by 94% of the population. Interestingly, 60% of those
tortillas are processed in small stores called tortillerias [1]. There are three main levels of
production and distribution of corn tortillas: 1) small commercial scale (S) (tortillerias), 2)
medium commercial scale or supermarkets (M), and 3) large commercial scale or samples
packed in plastic bags (L). Many aspects of the tortilla production still depend on the operator’s
expertise such as dough input, mixing, shaping, baking and other process parameters. The
study [2] revealed critical sensory attributes and gave a look into how Mexican consumers
weighted characteristics such as appearance, plasticity, chewiness, taste, and overall, liking
when making purchasing decisions about corn tortillas. With the necessity of more in-depth
information regarding the control process of corn tortilla production, the aim of this
investigation is to use a computer vision framework, to automatically determine the quality of
corn tortillas. The computer vision system is trained using the selected features obtained from a
panel of sensory analysis to classify automatically a tortilla.

MATERIALS & METHODS

A total of 750 corn tortillas were evaluated representing each level of corn tortilla production
(S, M, L), which represent a wide range of desirable and undesirable visual appearance
characteristics of corn tortillas. These characteristics contributed to overall visual appearance
depending on the origin production. In order to gain an understanding of the consumers’
preferences, a questionnaire was designed to establish which attributes were relevant to the
visual appearance of the samples. Mexican consumers N=100, who regularly ate corn tortillas,
answered the questionnaire. Colour was the most important attribute (around of 70%) while the
homogeneity of the borders or contours was the least important attribute to consumers (around
40%). Then, the samples were shown randomly to ten trained panellists and each one evaluated
depending on the three production levels (S, M, L). 100% of the samples were classified
correctly by the panellists. They determined five sub-classes according to the 5-point hedonic
scale (1: like extremely, 3: neither dislike nor like, 5: dislike extremely) as shown in Fig. 1.
Representative tortilla images were collected with a computer vision system similar to the one
described in [4] and were segmented according to that described by [3]; features from each
image were extracted and analyzed in order to select only those features that were relevant to
the classification tasks. Finally, 54 geometric and 2270 colour features were extracted. This
extraction and the subsequent classification of production levels and subclasses, was performed
using algorithms such as sequential forward selection (SFS), among others.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1753
 Figure 1: Gallery of corn tortilla images in
their three production level (classes) and
five sub-classes

RESULTS & DISCUSSION

75% of the samples of each class were
randomly chosen to perform the
feature selection. Results obtained in
the two mentioned classification parts:
(i) determination of hedonic scale (sub
class), and (ii) determination of production level (class), are discussed below. Hedonic scale
classification (sub-classes): Five data sets were built, one for each class. Forty features for each
data set using the SFS were selected. The performance was evaluated using cross-validation.
For m = 15 extracted features, our classifier had very high performance (average performance
Ș=0.95). In overall for this case, the sub-classes given by our classifier will be the same as the
ones given by the sensorial panel. For the second classification, two sets of features selected
using SFS were analyzed. Set-1 consisted on the best ten features selected from the 2324
extracted features. Set-2 contained the best 10 features chosen from the 64 features selected
from the hedonic scale classification. The performance of the classification using the
Mahalanobis classifier and the selected features for Set-1 and Set-2 were validated using cross-
validation. It is evident that Set-1 achieved a higher performance (Ș=0.995) than Set-2 although
this one also had a very high performance (Ș=0.96).

CONCLUSION
The method was tested in three different tortilla production levels with tortillas of five different
hedonic sub-classes yielding a performance of 95% in accuracy and 2770 features geometry
and color were extracted for classification. It is possible that the proposed approach opens new
possibilities not only in the field of automated visual inspection of tortillas but also in other
similar food products.

REFERENCES
[1] Ayala-Rodríguez, A., Gutiérrez-Dorado, R., Milán-Carrillo, J., López-Valenzuela, S.M., R.J., Valdez-
Ortiz, A., Paredes-López, O., Reyes-Moreno, C., 2009. Nixtamalised flour and tortillas from
transgenic maize (zea mays l.) expressing amarantin: technological and nutritional properties. Food
Chemistry 114 (1), 50–56.
[2] Herrera-Corredor, J., Saidu, J., Khachatryan, A., Prinyawiwatkul, W., Carballo-Carballo, A., Zepeda-
Bautista, R., 2007. Identifying drivers for consumer acceptance and purchase intent of corn tortilla.
Journal of Food Science 72 (9), S727–S730.
[3] Mery, D., Pedreschi, F., 2005. Segmentation of colour food images using a robust algorithm. Journal
of Food Engineering 66 (3), 353–360.
[4] Pedreschi, F., Mery, D., Mendoza, F., Aguilera, J., 2004. Classification of potato chips using pattern
recognition. Journal of Food Science 69 (6), E264–E270.

1754
Effect of Microwave Blanching on Acrylamide Content and Quality Attributes of French

Fries

Sezin Tutaa, Koray Palazoglua, Vural Gökmenb

a
Department of Food Engineering, Mersin,Turkiye(sezintuta@gmail.com,
koray_palazoglu@yahoo.com)
b
Department of Food Engineering, Ankara, Turkiye (vgokmen@hacettepe.edu.tr)

INTRODUCTION
Fried potato products are among the foods with the highest levels of acrylamide due to the high
level of acrylamide precursors naturally present in potato. Therefore, reducing acrylamide level
of fried potato products such as French fries is important. Blanching by electromagnetic energy
presents advantages over conventional blanching by reduction of process times, energy and
water usage and improvement of product quality [1,2]. The objective of this study was to
employ microwave blanching (instead of conventional blanching) during manufacturing of
frozen par-fried potato strips and investigate its effect on acrylamide content and quality
attributes (texture, color, oil content) of French fries.

MATERIALS & METHODS

Microwave blanching was performed by immersing the potato strips in boiling water and
microwaving for 1, 2, 3, and 4 min at 900 W. Blanched potato strips were then dried (5 min at
70 ºC), parfried (1 min at 180 ºC), cooled (20 min at 4 ºC) and frozen (2h at -30 ºC). Control
samples were prepared by two step blanching procedure ( 3 min at 80 ºC and 20 min at 65 ºC)
and all other steps were the same. Sensorial analysis (thoroughly cooked potato strip with a soft
interior and a crispy outer crust) were done to determine frying times for final frying at 180 °C.
Frying times for the control and 1, 2, 3 and 4 min microwave blanched samples were 5, 4, 3.5,
3 and 2.5 min, respectively. Color measurement was done by digital image analysis, Texture
Analyzer was used for texture measurement and oil content was determined using Soxhlet
method. An LC-MS method was used for acrylamide analysis.

RESULTS & DISCUSSION

Acrylamide reduction was % 18, % 48, % 71 and % 79 for blanching 1, 2, 3, 4 min with
microwave, respectively (Figure 1). It has been reported that microwave pre-treatment to
potato strips reduce the frying time resulting in a decrease in acrylamide level. Acrylamide
formation was reported to further decrease with increasing microwave application time [3,4].
In this research in contrast to conventional method potato strips were heated volumetrically
during microwave blanching resulting in a greater degree of cooking. As a result frying time
was found to decrease in comparison to control. It was observed that the longer the microwave
blanching time, the lower the surface temperature during frying , and hence less acrylamide
formation. Since potato tissue becomes more permeable upon microwave pre-treatment, less

11th International Congress on Engineering and Food - Athens, Greece, 2011 1755
resistance to mass transfer takes place during frying. This results in a moisture transfer from
the interior to the surface at a greater rate limiting temperature increase in the surface region.

Acrylamide, ng/g
800
600
400
200
0
Control MW1 MW2 MW3 MW4

The samples that were microwave-blanched for 3 min were the most comparable to the control
in terms of quality attributes of the final product (Table 1). Although a*, b* values were
significantly different in comparison to control (p<0.05), when visually evaluated all samples
appeared to be comparable in terms of color.

Table 1. Quality attributes of control and microwave blanched samples (mw_1,2,3,4).

Blanching
Color values Oil Content Fmax
time
(%) (N)
(min) L* a* b*
Control 83.68±1.16 -17.29±0.22 45.65±1.14 10.06±0.08 3.52±0.22
Mw_1 83.39±0.59 -14.03±0.69 54.50±3.30 7.85±0.97 2.81±0.15
Mw_2 84.27±1.80 -19.29±0.76 57.11±2.40 11.63±2.38 2.60±0.29
Mw_3 84.13±0.80 -19.64±0.87 60.56±2.55 13.17±3.16 2.73±0.50
Mw_4 85.83±1.11 19.66±0.13 59.19±3.24 10.01±0.56 1.91±0.40

CONCLUSION
In this study, frying time was reduced by microwave blanching while production of frozen
potato strips, and the effect of this application on acrylamide level and quality attributes of
French fries was determined. Acrylamide level was significantly reduced for 3 min (% 71) and
4 min (% 79). The method modification also shortened blanching times. Therefore, the results
of this study may be useful in industrial production of French fries. However, the effect of
microwave power should be investigated and optimization of blanching time should be
performed for industrial-scale microwave systems.

REFERENCES
[1] Ponne, C. T., Baysal, T., Yuksel, D. 1994. Blanching Leafy Vegetables with Electromagnetic Energy.
Journal of Food Science, 59(5), 1037-1041. [2] Ramesh, M. N., Wolf, W., Tevini, D., Bognar, A. 2002.
Microwave Blanching of Vegetables. Journal of Food Science, 67 (1), 390-398. [3] Erdo÷du, S. B.,
Palazo÷lu T. K., Gökmen V., ùenyuva H. Z., Ekiz H. ø. 2007. Reduction of Acrylamide Formation in
French Fries by Microwave Pre-cooking of Potato Strips. Journal of the Science of Food and Agriculture.
87: 133-137. [4] Tuta, S., Palazo÷lu, T. K., Gökmen, V. 2010. Effect of Microwave Pre-Thawing of
Frozen Potato Strips on Acrylamide Level and Quality of French Fries. Journal of Food Engineering. 97,
261–266.

1756
 Effects of Application of Tranglutaminase in Wheat Proteins During the Production of Bread.

Elisena Ap. Guastaferro Seravallia, Antonia Miwa Igutia, Inês Ap. Santanaa , Flavio Finardi Filhob

a
Maua Institute of Techonology-School of Engineering-Chemical and Food Engineering, Sao Caetano do Sul, Brazil
(elisena@maua.br)
b
University of Sao Paulo, Sao Paulo, Brazil

INTRODUCTION
The characteristics of proteins are one of the main parameters that affect the quality of wheat flour and
therefore the breadmaking quality. Some authors reviewed the structural models for the formation of the
dough and discussed chemical interactions that could influence the structure, with emphasis on reactions
of disulphide. In a accordance with researcher a protein of insoluble matrix is an essential prerequisite for
the formation of a cohesive dough; a quantity of insoluble protein is essential to form a continuous protein
phase in the presence of starch and water. Gerrard et al. [1] were the first researchers who used the
MTGase in wheat-based products. They suggested that the enzyme could have beneficial effects during
the manufacture of bread comparable to those produced by traditional oxidants improvers. The aim of this
study was to evaluate the use of transglutaminase as an improver agent in the bread and dough, and
compare the functional and molecular effects with the action exerted by traditional improver in the
protein fractions of glutenins, the raw dough, the dough after fermentation and the bread.

MATERIALS & METHODS

The procedure of sequential extraction, fractionation by solubility Osborne, was used with few
modifications. The protein content of each fraction was determined by Kjeldahl (factor=5.7). After
extraction of gliadins, the residue was used for extraction of glutenins. The glutenins were extracted at 20
°C with tetraborate buffer 0.05 mol/L (pH=8.5), 2-mercaptoethanol 2% (v/v), glycine 1 g/L and urea 6
mol/L, adapted by Larré et al. [2]. To avoid glutenin subunits repolymerization during the
chromatographic analysis, it was made the alkylation of these subunits [3]. The chromatographic fractions
of proteins were made using an HPLC system: Varian, a column Chromspher C8, 250 x 4.6 mm, particle
internal 5 Pm and UV visible detector 9050. The elution system used: A) Trifluoroacetic acid
(TFA)(0.1%,v/v), B) Acetonitrile (ACN)+TFA (99.9/ 0.1%, v/v), varying the gradients.

RESULTS & DISCUSSION

Moisture and protein for the doughs were evaluated before fermentation (M1), after 150 minutes of
fermentation (M2) and the bread after baking. All samples were removed during the manufacturing
process of bread. The doughs and bread Zero, bread Control and bread MTGase were analyzed (Table 1).
The results of the table 1 showed that the amounts of protein residues in M1 of the doughs are smaller
than those found in M2. These significant differences (p<0.05) can be observed both for the bread Zero
(2.3% to 3.0%) and bread MTGase (3.80% to 5.90%). However, the difference between the amounts of
protein samples of bread MTGase is much larger and can be explained by the presence of the enzyme that
can promote the polymerization of the subunits during the 150 minutes of rest dough. These results are
consistent with those found by some authors who worked with different methods to verify the presence of
protein polymers in residue.
After the separation of gliadins, the residue was used for extraction of glutenins. Below, the
chromatographic profiles from production of bread Zero and bread MTGase (Fig.1, 2).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1757
 Table 1-Levels of protein fractions of dough and bread(i).
Albumins/Globulins Gliadins Glutenins Residues Total Protein
M1(Zero) 1,44 r 0,01 2,84 r 0,06 ª,b
4,00 r 0,02 c
3,80 r 0,05 c
12,4
M2( Zero) 1,38 r 0,09 2,95 r 0,09a 2,23 r 0,01d 5,90 r 0,01d 12,8
bread Zero 1,30 r 0,01 2,85 r 0,03ª,b 2,2 r 0,3d 5,04 r 0,07e 11,6
M1(MTGase) 1,56 r 0,01 2,8 r 0,2 b,c
5,0 r 0,1 b,e
2,3 r 0,2f 12,0
M2(MTGase) 1,6 r 0,1 2,75 r 0,01c 4,10 r 0,06c 3,0 r 0,1g 12,0
bread MTGase 1,85 r 0,06 2,95 r 0,02 a
5,15 r 0,02 e
1,7 r 0,1h 11,76
(i) Means (5 determinations) with different letters in the same column differ statistically (p<0.05) by Duncan test.

Figure 1- Chromatographic profiles of glutenins from M1, M2, bread Zero. I=HMW-GS; II=LMW-GS.

Figure 2- Chromatographic profiles of glutenins from M1, M2, bread MTGase. I=HMW-GS; II=LMW-GS

The differences between the profiles of the flour, the doughs M1 and M2 for the bread Zero and bread
MTGase were clearly in high molecular weight glutenin subunits (HMW-GS), the least hydrophobic. For
the profiles of the doughs of bread Zero it was observed that there was a decrease in peak height for the
HMW-GS, M1 to M2, which may mean that during the rest of the dough decreased the soluble glutenins,
or also an increase of non-extractable glutenins, in the residue. Work performed by Caballero et al. [3],
showed that the addition of MTGase improved the functionality of the flour proteins through the
formation of large polypeptides resulting in decreased solubility.

CONCLUSION
We conclude that the chromatographic analysis revealed the influence of the enzyme in glutenins, but not
in gliadins. The rest of the dough reduced the extractability of glutenins, and the doughs containing the
enzyme reduction was more pronounced.

REFERENCES
[1] Gerrard J.A., Fayle S.E., Wilson A.J., Newberry M.P., Ross M. & Kavale S. 1998. Dough properties and crumb
strength of white pan bread as affected by microbial transglutaminase. Journal of Food Science, 63 (3), 472-475.
[2] Larrè C., Nicolas Y., Desserme C., Courcouxt P. & Popineau Y. 1997. Preparative separation of high and low
molecular weight subunits of glutenin from wheat. Journal of Cereal Science, 25, 143-150.
[3] Caballero P.A., Gomez M. & Rossel C.M. 2007. Improvement of dough rheology, bread quality and bread shelf-
life by enzymes combination. Journal of Food Engineering, 81, 42-53.

1758
 Agrophysical methods to determine bioenergetic status of agricultural products

Józef Horabik, Piotr Baranowski, Jerzy Tys

Institute of Agrophysics Polish Academy of Sciences, Lublin, Poland (jhorabik@ipan.lublin.pl)

INTRODUCTION
Detection of mechanical defects is very important in the systems of quality inspection of apples. Existing
automatic apple sorting systems still possess insufficient precision in detecting bruises, especially early
ones. In spite of the fact that bruising is the reason for rejecting the highest number of fruit in sorting
lines, the manual sorting method is still used (Leemans et al., 2004; Xing et al., 2007). In recent years
visual sorting systems were applied for detecting mechanical defects. They perform a multispectral
analysis of colour and use advanced procedures of image processing and analysis, including neural
networks, principal component analysis, etc. (Lu et al., 1999; Peng and Lu, 2006). Because of some
shortages of existing methods for early apple bruise detection, a growing interest is observed in
incorporating multi range non-destructive sorting methods. In this paper the comparison of hyperspectral
imaging (400-2500 nm) and thermal imaging (3500 -5000 nm) is presented.

MATERIALS & METHODS

Before hyperspectral and thermal imaging investigations ‘Jonagold’ and ‘Golden Delicious’ apples
(Malus domestica Borkh. The following imaging spectrographs were used in the study: the VNIR
ImSpector V10E imaging spectrograph (400-1000 nm) and the SWIR N25E 2/3” imaging spectrometer
(1000 nm – 2500 nm) by SPECIM, Finland. To reduce the dimensionality of data sets, to segregate noise
components from images and especially to produce uncorrelated output bands for which bruise
discrimination would be possible two methods were used. The first one was the Principal Components
Analysis (PCA). The second studied method was Minimum Noise Fraction Transform (MNF). An active
thermography system was designed, consisting of a thermographic camera, excitation source, and a
system for controlling the heat pulse time as well as registration parameters and external conditions in the
thermostated laboratory. In the study, a thermographic camera SC7600 (FLIR Systems, Inc., USA) was
used. It is sensitive in the spectral range of 3-5μm. Two approaches to active thermography were applied
in this study: the pulsed-phase thermography (PPT) and the lock-in thermography (LIT).

RESULTS & DISCUSSION

Principal component analysis of the apples of two studied varieties revealed its usefulness for extracting
the most relevant information from multiple band imagery gathered in a few component images. The use
of the Minimum Noise Fraction Transform (MNF) made the information about the bruise localisation and
composition even more readable. For VNIR range of wavelengths the most suitable components for
bruise detection were MNF4, MNF5 and MNF6 (Fig.1), whereas for SWIR bands, the components which
were the most independent from uneven illumination effects and could be used for bruise detection were
MNF4 and MNF5. In Fig. 2 the results of this analysis are presented for ‘Jonagold’ apple. In this figure,
thermogram marked with a) is the image of the apple before heat pulse or wave extinction (cold image).
Images in b) and c) are the results of lock-in analysis.
The lock-in ampligram is strongly influenced by reflections from illumination source. Therefore, the
phase analysis, which is free of this influence is preferable for defect recognition. The images in d), e) and
f) are phasegrams for various phase shifts of the same fruit obtained by pulsed-phase thermography.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1759
 MNF1 MNF2 MNF3

MNF4 MNF5 MNF6

Figure 1. Minimum Noise Fraction Transform (MNF) scores images for the VNIR wavelength range

a) thermal image without b) lock-in ampligram c) lock-in phasegram

heating

d) pulsed- phase (phasegram 1) e) pulsed- phase (phasegram 2) f) pulsed- phase (phasegram 3)

Figure 2. Scores of active thermography (3-5 Pm) of ‘Jonagold’ apple

CONCLUSION
The whole studied spectrum range (400 nm- 5000 nm) is useful for detecting mechanical defects such as
bruise. Hyperspectral analysis of VNIR and SWIR wavebands can be effectively performed by
application of principal component analysis (PCA). Even better results are obtained by the use of
minimum noise fraction transform (MNF) which components could be preferable for image segmentation
purpose. Thermal imaging (3000 nm -5000 nm) is useful for bruise recognition when active approach
(lock-in or pulsed-phase) is applied.

REFERENCES
[1] Leemans, V., Magin, H. & Destain, M.F. 2002. On-line fruit grading according to their external quality using
machine vision. Biosyst. Eng. 83, 397–404. [2] Lu, R., Chen, Y.R. & Park, B. 1999. Hyperspectral imaging for
detecting bruises in apples. ASAE Annual International Meeting. Paper No. 99-3120. Toronto, Canada, 18–21 July. [3]
Peng, Y. & Lu, R. 2006. An LCTF-based multispectral imaging system for estimation of apple fruit firmness. Part II.
Selection of optimal wavelengths and development of prediction models. Trans. ASAE 49, 269–275. [4] Xing, J.,
Jancso, P., De Baerdemaeker, J. 2007. Stem-end/calyx identification on apples using contour analysis in multispectral
images. Biosystems Engineering 96, 231–237.

1760
 Separation between high and low quality coffees by FTIR-ATR

Adriana S. Francaa,b, Ana Paula Craigb, Leandro S. Oliveiraa,b

a
Departamento de Engenharia Mecânica, Universidade Federal de Minas Gerais (UFMG), Av. Antônio
Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil (adriana@demec.ufmg.br)
b
Programa de Pós-Graduação em Ciência de Alimentos, UFMG, Belo Horizonte, MG, Brazil

INTRODUCTION
The presence of defective coffee beans depreciates the quality of the coffee beverage
consumed worldwide. The intrinsic defects (sour, black and immature) are the ones that, when
roasted, contribute the most to the depreciation of the coffee beverage quality. Colour sorting is
the major procedure employed for separation of defective and non-defective coffee beans prior
to roasting. However, such procedure is not efficient for separation of immature beans. Thus,
the objective of the present study was to evaluate the feasibility of employing FTIR for
separation between high quality (non-defective) and low quality (defective) coffee beans.

MATERIALS & METHODS

Arabica green coffee samples were comprised of coffee beans obtained from different
cooperatives located in Minas Gerais State, Brazil, that were rejected by color sorting
machines. Black, sour (separated into light and dark coloured), immature and non-defective
beans were manually picked to constitute separate sampling lots and ground to a particle
diameter of 0.42 mm. A Shimadzu IRAffinity-1 FTIR Spectrofotometer (Shimadzu, Japan)
with a DLATGS (Deuterated Triglycine Sulfate Dopedwith L-Alanine) detector was used in
the measurements that were all performed in a dry atmosphere at room temperature
(20 ± 0.5 °C). A horizontal ATR sampling accessory (ATR-8200HA) equipped with ZnSe cell
was employed. All spectra were recorded within a range of 4000–700 cmí1 with a 4 cmí1
resolution and 20 scans. Spectra treatment consisted of baseline correction and normalization.

RESULTS & DISCUSSION

PCA analysis of the ATR reflectance spectra, employing baseline correction and normalization
is displayed in Figure 1. Analysis was based on a 54 x 1188 data matrix assembled so that each
row corresponded to a sample and each column represented the spectra data at a given
wavelength. The two first components accounted for 76% of the total sample variance. The
first component provided separation of the evaluated samples into two major groups: non-
defective/light sour (positive PC1) and black/dark sour/immature (negative PC1). Evaluation of
the loadings plot (not shown) indicated that the spectral ranges that presented the highest
influence on PC1 values in association with the black/dark sour/immature group were the
following: 1482-1554, 1776-1797 and 3020-3100 cm-1. The only significant band that can be
observed in the ATR spectra in those ranges is the one at 1534 cm-1. It was also present in the
spectra obtained by Lyman et al. [1] for aqueous extracts of roasted coffee, regardless of
roasting conditions, although no identification was attempted. In the case of PCA based on the
first-derivative of the spectra (not shown), the first and second principal components accounted

11th International Congress on Engineering and Food - Athens, Greece, 2011 1761
for 28.6 and 12.6% of the total sample variance, respectively. No separation could be observed.
The results presented in Figure 1 confirm that separation between high (non-defective) and low
quality (black, immature and dark sour) coffees can be accomplished by FTIR-ATR analysis.
Although light sour beans could not be separated from non-defective ones, such separation
could be easily performed by bi-chromatic colour sorting. Given that the main problem with
colour sorting is the separation of immature and non-defective beans, colour sorting could be
employed as a first step to eliminate sour and black beans and thus FTIR-ATR could be
employed for separation between immature and non-defective coffees.

Figure 1. PCA scores scatter plot of FTIR-ATR spectra submitted to normalization and baseline
correction (PC1 vs. PC2). non-defective; immature; sour (light); sour (dark); black.

CONCLUSION
The feasibility of employing FTIR as a methodology for the separation between low quality
(defective) and high quality (non-defective) coffees was evaluated. PCA results based on
normalized ATR-FTIR reflectance spectra indicated separation of the samples into two major
groups: non-defective/light sour and black/dark sour/immature. The preliminary results
obtained in the present study confirm that FTIR analysis presents potential for the development
of an analytical methodology for separation between defective and non-defective coffee beans.
Further studies will be conducted employing a larger set of samples in order to develop
predictive models. The methodology will be also tested for roasted coffee samples.

ACKNOWLEDGEMENTS
The authors acknowledge financial support from the following Brazilian Government
Agencies: CNPq and FAPEMIG.

REFERENCES
[1] Lyman, D.J., Benck, R., Dell, S., Merle, S. & Murray-Wijelath, J. 2003. FTIR-ATR analysis of
brewed coffee: effect of roasting conditions, Journal of Agricultural and Food Chemistry, 51, 3268-3272

1762
 Effect of temperature on biospeckle activity in apples

Andrzej Kurendaa, Anna Adamiaka, Artur Zduneka

a
Department of Microstructure and Mechanics of Biomaterials, Institute of Agrophysics PAS
DoĞwiadczalna 4, 20-290 Lublin 27, Poland

INTRODUCTION

Optical techniques are fast developing group of methods of food products quality evaluation, because of the
speed and precision of measurement as well as the non-destructive character. Biospeckle is another optical technique,
although less known, that could be used to this purpose.
Previously conducted experiments on biological materials have demonstrated the usefulness of biospeckle
technique in issues such as:analysis of maturation and bruising of fruits and vegetables [1], monitoring of apple shelf
life [2]. However it is still unknown what kind of biological processes are at the basis of this phenomenon. It is
suspected that the biospeckle activity is caused by biological processes such as: cytoplasmic streaming, organelle
movement, cell growth and division during fruit maturation and biochemical reactions, but processes such as Brownian
motion or diffusion should also be taken into account.
To test whether the biospeckle activity is caused by metabolic or physical processes, an experiment was made
which consists in recording the activity phenomena with decreasing temperature of the biological object. In the case of
living organisms, whose operation is based on enzymes, the rate of biological processes depends on temperature in a
characteristic non-linear way, unlike processes such as Brownian motion, which in the investigated temperature range
should change linearly. An example of that temperature-dependent biological process is the cytoplasmic streaming in
Characeae cells [3] which is suspected to be one of the biospeckle sources.

MATERIALS & METHODS

The device for biospeckle measurements was similar to that which was previously used by Zdunek et al.[2].
The experiment consisted in examination of the biospeckle activity in three variants of temperature: a constant
temperature of 24°C, a constant temperature of 4°C and at gradually lowering the temperature from 29 to 5°C.
Biospeckle obtained by illumination of fruits by He-Ne laser light were recorded and their activity was determined by
three methods of image analysis, involving assessment of the correlation coefficients between the first and successive
frames, contrast changes and moments of inertia of the co-occurrence matrix in subsequent frames in time.

RESULTS & DISCUSSION

The results of our experiments unequivocally indicate that all the methods of image analysis reveal a stability of
phenomena in a constant temperature and a decrease in the biospeckle activity with decreasing temperature. We also
observed some differences between the methods of assessment of the activity of the phenomenon studied. The highest
differences in the activity between extreme temperatures are visible in the method of testing the correlation coefficient,
and the lowest - in the method of contrast evaluation
Surface of apples placed in a 2°C, reached a minimum temperature of about 4°C after 4-5 hours of the
experiment. In additional experiments, it was found that despite 24 hours of storage at 2°C, apples did not reach the
ambient temperature which may be a side effect of metabolic reactions. As shown in Figure 4, the values of correlation
coefficient increase with decreasing temperature which means a decrease of biospeckle activity. The relationship
between temperature and the correlation coefficient is nonlinear and shows the best fit to the third degree polynomial
functions as confirmed by the coefficient of determination R2 of about 0,95. The chart also contains the standard
deviation for the biospeckle activity in different temperatures.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1763
 Phenomena related to the movement of substances and organelles in cells are probably the cause of coherent
light scattering and consequently the biospeckle activity. All processes of active transport are linked with the
consumption of energy present in the cells in the form of high-energy chemical bonds. So if the source of the
biospeckle are biochemical reactions occurring in a cell, the lowering the temperature of the


Figure 4. Biospeckle activity with temperature decrease, expressed as the inverse of the correlation coefficient.

object will cause decrease of biospeckle activity of the same nature as decrease in the rate of biochemical reactions,
which in the investigated temperature range is S-shaped [4].Given that the relationship between temperature and other
phenomenon that may affect the activity of biospeckle - Brownian motion - is linear [5], the relationship shown in
Figure 4 is more appropriate for the dependence temperature-biochemical reactions than for the temperature-Brownian
motion.

CONCLUSION

The nature of changes of biospeckle activity with temperature, shows that the main source of phenomena are
biochemical processes (which probably include intracellular transport processes) rather than physical processes.
Moreover, assuming that the biospeckle activity is caused by life processes, and taking into account the high sensitivity
of the method on the rate of these processes, we believe that it can be used to evaluate some parameters of fruits and
vegetables quality as well as in biological research.

REFERENCES
[1] Pajuelo, M., Baldwin, G., Rabal, H., Cap, N., Arizaga, R., Trivi, M., 2003. Bio-speckle assessment of bruising in fruits.
Opt. Lasers Eng. 40, 13–24.
[2] Zdunek, A., Frankevych, L., Konstankiewicz, K., Ranachowski, Z., 2008. Comparison of Puncture Test, Acoustic
Emission and Spatial-Temporal Speckle Correlation Technique as Methods for Apple Quality Evaluation. Acta Agrophys.
11(1), 303-315.
[3] Shimen, T., Yoshida, S., 1993. Analysis of temperature dependence of cytoplasmic streaming using tonoplast-free cells of
Characeae. Protoplasma. 176, 174-177
[4] Kotov, N.V., Baker., R.E., Dawidov, D.A., Platov, K.V., Valeyev, N.V., Skorinkin, A.I., Maini, P.K., 2007. A study of the
temperature dependence of bienzyme systems and enzymatic chains. Comput. Math. Meth. Med. 8(2), 93-112.
[5] Jia, D., Hamilton, J., Zaman, L.M., Goonewardene, A., 2007. The time, size, viscosity, and temperature dependence of the
Brownian motion of polystyrene microspheres. Am. J. Phys. 75(2), 111-115.

1764
 Implementation of DNA technology in a Greek dairy company: An overview

E. Beletsiotisa, D. Ghikasa, K. Kalantzia

a
DELTA FOODS S.A, Athens, Greece (vagbel@delta.gr, dimghi@ delta.gr , kelkal@ delta.gr )

INTRODUCTION
The knowledge and understanding of microbial ecosystem (micro-environment) of each production
facility is a necessity in order to avoid spoiling of the products and constitute an important part of
HACCP plan. The microbiological analyses, conducted by Quality Control (QC) department, at the
critical points of the production lines, gives information about the diversity and taxonomic identity
of the species that could contaminate the food at every stage of production [1]; [2]. Applications of
DNA technology in food industry are relatively new. The construction of a Molecular Biology
Laboratory (MBL) and the incorporation of molecular techniques, in the QC control scheme of a
dairy company, is an innovation for the Greek industry. The MBL operates providing auxiliary
support to the monitoring of the micro-environment for each of the manufacturing plants, according
to HACCP standards each one has set. Services provided include microorganism and arthropod
molecular identification, typing of microorganisms and GMO analyses in foodstuff and products.
Another aspect of MBL is to co-operate with QC and R&D, in projects and certain scientific needs.

MATERIALS & METHODS

MBL uses a variety of standard microbiological and molecular techniques. For microorganism
culturing a large number of microbial practices are followed according to ISO, IDF standards and
QC working instructions of each plant. Isolation of DNA, Polymerase Chain Reactions (PCR), Real
Time PCR (RT-PCR), sequencing and typing of microorganisms, GMO analyses are made
according standard molecular techniques and kit utilization. Resulted sequences analyzed with
Lasergene suite programs (DNASTAR) and then they were screened using BLASTn algorithm
(www.ncbi.nlm.nih.-gov/BLAST/).

RESULTS & DISCUSSION

Identification of microorganisms was made, using sequencing information, obtained from 16S
rDNA gene for bacteria and from 28S rDNA region for fungi.Total number of analyses conducted
for strain identification from 2008 until 2010 was 2850 for bacterial species and 1741 for fungal
species from specimens provided by QC of six different plants. Differences in number of
microorganisms identified for each plant was due to the kind of product, volume of production and
monitoring scheme. Although, all plants processed milk and milk products, each one has its unique
micro-environment. Differences, in percentages of bacterial taxinomical orders identified, vividly
indicate different manufacturing practices, processes and adaptation of microorganisms in each
facility. Data obtained from identification of microorganisms, ecology, specific needs for nutrients,
growth rates and conditions (temperature, humidity) and isolation frequency in the plant
environment, allowed the creation of “micro-bank”. The identity of microorganisms allowed us to
obtain a better image of the micro-environment of the plants, to decide and apply appropriate
corrective measures, regarding changes in the production area and machinery [2].
Other services provided are related to Integrated Pest Management (IPM) of each plant. The
laboratory developed and applied a molecular technique for the identification of arthropods in the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1765
order/genus level. In one year application 82 analyses have been made and interesting conclusions
have been emerged. Arthropods, that have been identified from plants, participating in that program,
belonged mainly to class Insecta. Geographical position and specific environmental conditions
applied in each manufacturing plant are among the factors that contribute substantial to the
differences, observed in insects order and genus level among the different plants. An “insect-bank”
was constructed, containing data regarding the ecology of the insects identified, the ethology, the
isolation sites inside the plants, while new data continuously enrich the data bank. These data are
taking into consideration from the personnel of Q.C when the IPM control scheme is re-evaluated
and corrective measures are taken if needed.
Another aspect of MBL is to work with QC and R&D, in projects and certain scientific needs.
Information related to species and strain level of microbial strains for two R&D projects regarding
yoghurt starter cultures and an ongoing typing project for one of our plants have been made.
MBL has a supportive role in the surveillance plan for GMO presence in foodstuff and company’s
final products. Analyses were made with RT-PCR techniques, for qualitative and quantitative
identification of soya and GMO crops harboring NOS-terminator and 35S promoter insertions and
has the molecular tools, to identify the majority number of soya and maize modifications of crop
varieties, approved for food use in Europe [3]. In a three year period the MBL has analyzed 362
samples for presence/absence of crop and soya GMO modifications and last year, up to 126
different analyses have been conducted, for the detection of maize GMO events. All the results were
negative for the presence of GMO material. These results are in accordance with results obtained
from external laboratory tests indicating that foodstuff and company’s final products are “GMO
free”.

CONCLUSIONS
This study is unique as it describes examples of the implementation of DNA technology in a Greek
dairy company.
a. Routinely, microorganisms are identified using molecular techniques, with high speed and
accuracy. That allowed the construction of a micro-bank, specific to the micro –environment
of each facility. Knowledge of that kind, allows the company to take appropriate preventive
measures assuring high level of safety in the final products.
b. Molecular identification of arthropods, which to our knowledge, is unique application among
Greek industries, provides accurate information for the construction of an insect-bank and
optimization of IPM scheme for each plant.
c. Participation in “in house” research projects, resulting to novel approaches and contributing to
a higher level of scientific information, regarding microbiological aspects.
d. GMO analyses, for all known soya and maize genetic events, resulting to a higher confidence
level of GMO free policy, of the company.

REFERENCES
[1] Fleet G.H. 1999. Microorganisms in food ecosystems. International Journal of Food Microbiology, 50, 101-117.
[2] Marco M.L. & Wells-Bennik M.H. 2008. Impact of bacterial genomics on determining quality and safety in the
dairy production chain. International Dairy Journal, 18, 486-496.
[3] Batista R. & Oliveira M.M. 2009. Facts and fiction of genetically engineered food. Trends in Biotechnology. 27(5),
277-286.

1766
 Sensorial Characteristics of Goat Milk Cheeses Made From Ultra High-Pressure

Homogenization-Treated Milk

B. Juan, J. M. Quevedo, B. Guamis, V. Ferragut, A. J. Trujillo

Centre Especial de Recerca Planta de Tecnologia dels Aliments (CERPTA), XaRTA, TECNIO, MALTA
Consolider, Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, 08193
Bellaterra (Spain)

INTRODUCTION
Ultra-high pressure homogenization (UHPH) is one of the most promising alternatives to
traditional thermal treatment of food preservations and diversification. UHPH has been used
for the stabilization of emulsions and to inactivate harmful bacteria in milk [1]. However, little
research exists on the sensory characteristics of milk cheeses made from UHPH-treated milk.
The aim of the current work was to study the effect of UPHP in goat milk at a pressure of 200
MPa on the sensorial characteristics of goat milk cheeses (UH-cheeses). The results obtained
were compared with those from cheeses produced by conventional homogenized-pasteurized
(18+2 MPa, 72ºC for 15 s, PH-cheeses) and heat-pasteurized (72ºC for 15 s, PA-cheeses)
milks.

MATERIALS & METHODS

Ultra-high pressure homogenization was carried out by subjecting milk to single stage UHPH
(200 MPa) using a Stansted high-pressure homogenizer (model FPG11300, Stansted Fluid
Power Ltd., Essex, UK) at an inlet temperature of 30 ± 1°C. Two-stage homogenization (18
MPa plus 2 MPa) and pasteurization (72ºC for 15 s) of raw milk were carried out with a Niro
Soavi homogeniser (model X68P, Parma, Italy) and a Finamat heat exchanger (model
6500/010, Gea Finnah GmbH, Ahaus, Germany), respectively. Three independent productions
of cheese were carried out in the Centre Especial de Recerca Planta de Tecnologia dels
Aliments (CERPTA) at the Universitat Autònoma de Barcelona.
Cheeses were analysed for pH, total solids (TS) [2], moisture content (100 – TS), total nitrogen
(TN) [3] and protein (TN x 6.38). Instrumental texture was evaluated by uniaxial compression
test using a TA-TX2 Texture Analyser (State Micro system, Surrey, UK) as described by [4].
Sensory assessments of 60 day-old cheese samples were performed by a panel of 10 members
from the CERPTA. Hardness, springiness, granular, sticky and watery were used for described
the cheese texture. Intensity of flavour, aroma and off-flavour also were evaluated. Panellists
marked responses on a 9-point intensity scale were pasteurized milk cheeses (PA) were used as
a control (0 = no differences with control, ±1 = minimal differences, ±2 = noticeable
differences, ±3 = considerable differences, ±4 = very considerable differences). Negative (-) or
positive (+) indicates lower or greater perception related to control sample. Panellist also
described the preference order of cheeses.
Analysis of variance (ANOVA) was performed by SPSS Win version 17.0 (SPSS Inc.,
Chicago, IL). Student-Newman-Keuls test was used for comparison of sample data.
Evaluations were based on a significance level of P < 0.05.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1767
RESULTS & DISCUSSION
No differences in pH values were observed between cheeses at 1 or 60 days of ripening. At 60
days of ripening UH-cheeses showed the highest fracture stress and fracture strain, becoming
the firmest and the most deformable cheeses, agreeing with the sensorial analysis described by
the panellists. The higher values of moisture content found in UH-cheeses would contribute to
a cheese that deformed more easily.

Table 1. Texture parameters of PA, PH and UH-cheeses at 1 and 60 days of ripening.

Fracture stress (x10 kPa) Fracture strain (-)
Days 1 60 1 60
PA 3,47 ± 0,62b 38,49 ± 2,95b 0,90 ± 0,17a 0,27 ± 0,05c
PH 2,58 ± 0,44c 36,92 ± 6,32b 0,96 ± 0,05b 0,31 ± 0,03b
UH 5,65 ± 0,78a 49,01 ± 7,30a 0,43 ± 0,05c 0,40 ± 0,04a
abc
Means within the same column followed by different superscript are significantly different (P<0.05)

Panellists described UH-cheeses grainier and less watery than PA-cheeses. The release of
water inside the mouth during mastication would be linked to the structure of the cheese rather
than the water content. The homogenization enhances casein-casein interactions resulting in a
tight protein matrix [5] increasing the entrapped water in UH-cheeses and resulting described
as less watery cheeses in spite its high moisture content. PH-cheeses were qualified less firm
than PA-cheeses. However, panellist described PH-cheeses like the springiest, pastiest, less
grainy and most watery cheeses. The descriptive analysis showed that no statistical differences
were found on the mean scores of flavour and aroma intensities in cheeses. Results of the
preference test showed that panellist preferred PA-cheeses on the first position, followed by
UH-cheeses. PH-cheeses were evaluated in the last position.

CONCLUSION
The treatment of UHPH in goat milk at a pressure of 200 MPa produced slowly differences in
the texture characteristics of cheeses, increasing the firmness and granularity of cheeses.
However, these differences were not evaluated as defects by the panellists. In addition, UHPH-
treatment had any important consequences in flavour of aroma of cheeses, suggesting that it
could be used as an alternative to traditional thermal treatment to make goat milk cheeses.

REFERENCES
[1] Pereda, J., Ferragut, V., Quevedo, J.M., Guamis, B. & Trujillo, A.J. 2007. Effects of Ultra-High Pressure
Homogenization on microbial and physicochemical shelf life of milk. Journal of Dairy Science, 90, 1081-1093.
[2] IDF. 2004. Cheese and processed cheese-Determination of the total solids content (reference method). IDF
Standard 004:2004. International Dairy Federation, Brussels, Belgium.
[3] IDF. 2002. Milk and milk products-Determination of nitrogen content. IDF Standard 185:2002/ISO 14891.
International Dairy Federation, Brussels, Belgium.
[4] Juan, B., Trujillo, A.J., Guamis, B., Buffa, M. & Ferragut, V. 2007. Rheological, textural and sensory
characteristics of high-pressure treated semi-hard ewes’ milk cheese. International Dairy Journal, 17, 248-254.
[5] Zamora, A. 2009. Ultra-high pressure homogenization of milk: effects on cheese-making. Doctoral thesis.
Universitat Autònoma de Barcelona.

1768
 User-friendly software predicting the microbial spoilage of emulsified acid foods

Stavros G. Maniosa, Antonis Psomas, Panagiotis N. Skandamis

a
Laboratory of Food QualityControl and Hygiene, Department of Food Science and Technology,
Agricultural University of Athens, Iera Odos 75, 118 55, Athens, Greece. (pskan@aua.gr)

INTRODUCTION
“Traditional Greek Salads” (TGS) are highly acidic, pourable appetizers which are
manufactured based on traditional recipes. Their low pH (3.6 – 4.5) in combination with salt
(1-2%) allow a shelf life of 2-3 months under refrigeration. However, temperature abuse during
distribution or retail and home storage may accelerate the microbial spoilage of these products,
especially during summer. A variety of kinetic models has been developed to predict the
growth of spoilage microorganisms in foods. These models are useful for the determination of
the shelf-life of such products. However, there is lack of models predicting the shelf-life of
weak acid acidified, RTE products which are being stored under refrigeration. Thus, the
development of a ‘generic’ model, which is able to correlate microbial growth with the initial
pH and weak acids concentration of such foods, is of high importance for assessing microbial
stability of similar products. The objectives of this study were; (i) to characterize the microbial
spoilage of three TGS, by correlating changes in microbial populations with sensory
evaluation, physicochemical changes, as well as changes in species diversity during isothermal
storage; (ii) to develop and validate a predictive spoilage model in the form of a user-friendly
software (i.e., tertiary model based on Visual Basic platform), which may provide predictions
on the microbial status and the remaining shelf life of the above products under dynamic chill
chain conditions and; (iii) to evaluate the applicability of a unified (generic-like) predictive
model of microbial spoilage based on the initial concentration of the acidulant (acetic acid) and
the initial pH of relevant products, regardless of differences in the recipe and the
manufacturing process.

MATERIALS & METHODS

Commercial samples of Pepper-based salad (PS), Fava beans-based salad (FS) and Eggplant-
based salad (ES) were stored under isothermal conditions (4 – 25oC) until occurrence of
spoilage. The populations of lactic acid bacteria (LAB) were primarily modelled through the
Baranyi model. Growth rates were further modelled as a function of temperature with a
polynomial model, based on storage temperature, the initial pH and undissociated acetic acid
concentration. The developed model was validated under dynamic temperature conditions in
four household refrigerators and measured LAB populations were compared with those
predicted by the model. Changes in pH, titratable acidity and organic acids concentration
(HPLC) as well as sensory characteristics were correlated with LAB population at the point of
organoleptic rejection, in order to define the spoilage level of LAB. In addition, LAB isolates
from each appetizer and storage temperature were characterized based on their SDS-PAGE
pattern and were further identified with 16s rRNA

11th International Congress on Engineering and Food - Athens, Greece, 2011 1769
RESULTS & DISCUSSION
The shelf-life of the products was defined as the time that organoleptic rejection occurred. This
coincided with LAB populations levels of 7.5-8 log CFU/g, drop of pH below 3.9 and increase
of NaOH consumption over 3 ml. The latter two may likely occur as a result of lactic acid
production which was observed in all appetizers or acetic acid which is a metabolic product of
hetero-fermentative LAB. Other organic acids such as succinic, malic and quinic displayed
mildly reduction whereas citric acid remained almost unchanged. These results where further
explained by the molecular analysis or the specific spoilage microorganisms (SSOs). Notably,
despite the large diversity in species at the beginning and early storage, Lactobacillus brevis
and L. plantarum dominated the microbial flora of the products at the onset of spoilage. This
SSOs diversity likely affected the dependency of ȝmax on storage temperature. The initial pH
and acidity of the products strongly influenced their shelf-life. Products with lower pH and/or
higher acetic acid content showed higher microbial stability, while in ES different treatments
on the added eggplant (with or without dipping in lactic acid) appeared to affect the growth rate
of LAB. In a further step, a unified-generic model which described the predicted maximum
specific growth rates of LAB in all weak acid acidified products, as a function of temperature,
the initial pH and the undissociated, weak acid concentration was assessed, and the predictions
demonstrated good agreement with the populations of LAB observed during storage under real
chill chain conditions (Fig. 1)

Figure 1 Predictions of the unified – generic model (—) correlated with the observed (z) growth of lactic
acid bacteria in fava beans salad, stored under two different time-temperature (---) profiles. An average ho
value of 1.13 was used for the simulations.

CONCLUSION
The results suggest that the potential of microbial spoilage of these products is highly
associated with their initial pH and potential temperature fluctuation, with a temperature
stability zone between 4 and 10oC. The proposed dynamic model may assist in the
management of such products during chill chain. It may also be used as a tool to characterize
the stability of emulsified acetic-acid based foods as a whole product category, based solely on
their initial acidulant concentration and pH levels.

1770
 Detection of fecal contamination on leafy greens by hyperspectral imaging

Sukwon Kang, Kangjin Lee, Jong-Guk Lim, Moon S. Kima

Rural Development Administration, Suwon, Korea

a
Agricultural Research Service, USDA, Beltsville, MD, U.S.A.

INTRODUCTION
It has been reported that several E.coli O157:H7 outbreaks were associated with shredded
iceberg lettuce and romaine lettuce [1]. The origin of the such contamination may from
irrigation water contaminated with wildlife feces [2]. These foodborne illness outbreaks drive
the food safety initiatives to save public health from pathogens associated with fresh produce.
Thus, development of automatic inspection system with fast and accurate has been required to
detect the bovine fecal contamination on fresh produce.
Hyperspectral imaging technique has been applied to food quality and safety to detect the
defects or fecal contamination on fruits and vegetables [3, 4] and find the microbial
contamination such as bacterial biofilms on food processing equipments [5].
From the hyperspectral imaging technique, the fluorescence imaging showed good detection
results for the spots of diluted fecal contamination on apples. Detection can be allowed by
contaminant constituents in feces such as chlorophyll a and its byproducts .
The objective of this study is to use a hyperspectral fluorescence imaging system to detect the
bovine fecal matter on leafy green vegetables such as romaine lettuce and baby spinach. Image
processing algorithm will be investigated to find the possibility of detecting the fecal
contamination spots on leafy green vegetables.

MATERIALS & METHODS

The prepared leafy vegetables are romaine lettuce (Lactua sativa L.) and baby spinach leaves
(Spinacia olerace L.) and they were obtained from a local market. Two or three leaves from
each head of lettuce were taken and washed for approximately 30 seconds. The each spinach
leaf was washed by the same method. The feces samples are obtained from Holstein cows
which are growing at the Dairy Operations Unit, Beltsville Area Research Center, Agricultural
Research Service, United States Department of Agriculture. Forty leaves of romaine lettuce
and forty leaves of baby spinach leaves were prepared for the fecal contamination. For each
fresh produce, twenty leaves were used the adaxial surface and another twenty leaves were
used for abaxial surface. Approximately 5-mm diameter fecal spot was applied on each lettuce
and baby spinach leaf surface using a spatula. The used hyperspectral line-scan imaging
systems has electron-multiplying-charge-coupled-device (EMCCD) camera, an imaging
spectrograph, lenses, a pair of ultraviolet (UV)-A lights at 365 nm providing near-uniform
illumination to the linear field of view (FOV). The image size for analysis was reduced to 60
(spectral) × 200 (spatial) pixels. Image acquiring program was developed by using MS Visual
Basic with the development kit provided by the EMCCD manufacturer. After hyperspectral
fluorescence images are acquired, image processing program was used to analyze the image
data using the Principal Component Analysis (PCA).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1771
RESULTS & DISCUSSION
The spectra of soil were similar to those of non-fluorescent black cloth. The emission maxima
for the romaine lettuce and baby spinach leaves were between 660 nm and 690 nm,
approximately, and this may be related to the intensity of chlorophyll a in the romaine lettuce
and baby spinach leaves. Compared to the emission peak for leafy green surface, the emission
peak for the fecal contamination spots are shifted toward the shorter wavelength. It was
distinguishable that each emission peaks for vein, inter-vein, and bovine manure.
PCA analysis results showed that the hyperspectral imaging system with the developed
detection algorithm could successfully detect the fecal contamination on the leaf surfaces and
differentiate the contaminated fresh produce from the uncontaminated ones. The result also
showed that the differences between two leaf surface (adaxial and abaxial), and two leaf
surface areas (inter-vein and vein) did not cause any difficulty to the detection of bovine fecal
contamination. However, it is necessary to develop the different PC band for the different fresh
produce for the higher detection accuracy. This benefit is essential for online application of the
algorithm by saving computation and operation time and effort.

CONCLUSION
In this study, the hyperspectral fluorescence line-scan imaging system was used for the
detection of fecal contamination on the leaf surface of fresh produce. The research indicated
that the principal component analysis method can be used for bovine fecal contamination
detection. The mask created by the waveband at the 692 nm could be used to distinguish the
objects from the background before obtaining the image. The threshold of 0.45 was applied to
the ratio image to mark the pixels with fecal contamination, and the 3 × 3 filter was then
applied to eliminating false positive pixels. The final binary images for certain principal
component showed that the algorithm could successfully detect all of fecal contamination spots
on the adaxial and abaxial surfaces of romaine lettuce and baby spinach.

REFERENCES
[1] FDA. 2006. Questions & Answers: Taco Bell E. coli O157:H7 lettuce outbreak. FDA, Washington,
DC.
[2] Armstrong, G.L., Hollingsworth, J. and Morris Jr., J.G. 1996. Emerging foodborne pathogens:
Escherichia coli O157:H7 as a model of entry of a new pathogen into the food supply of the
developed world. Epidemiologic Reviews, 18(1), 29-51.
[3] Kim, M.S., A. M. Lefcourt, A.M., Y. R. Chen Y. R. & Yang, T. 2005. Automated detection of fecal
contamination of apples based on multispectral fluorescence image fusion. Journal of Food
Engineering, 71(1), 85-91.
[4] Vargas, A.M., Kim, M. S., Tao Y., & Lefcourt., A. M. 2005. Detection of fecal contamination on
cantaloupes using hyperspectral fluorescence imagery. Journal of Food Science, 70(8), 471-476.
2005.
[5] Jun, W., Kim, M. S., Lee, K., Millner, P., & Chao, K. 2009. Assessment of bacterial biofilm on
stainless steel by hyperspectral fluorescence imaging, Sensing and Instrumentation for Food Quality
and Safety, 3(1), 41-48.

1772
 Detection of mushroom Virus X (MVX) infection in asymptomatic mushrooms using

FTIR microscopic imaging

Alvarez-Jubete La, Bonnier Fb, Byrne Hb, Grogan Hc, Frias JMa

a
Dublin Institute of Technology, School of Food Science and Environmental Health, Dublin, Ireland,
City, Ireland
b
Dublin Institute of Technology, FOCAS Research Institute, Dublin, Ireland
c
Horticultural Development Unit, Teagasc, Kinsealy, Dublin, Ireland

INTRODUCTION
Mushroom Virus X affects important traits associated with mushroom quality including colour
and appearance [1]. As a result, the spread of Mushroom Virus X to mushroom crops may
potentially result in devastating economical effects for mushroom growers and producers. To
prevent cross-contamination from occurring, it is essential than MVX infected crops can be
readily identified. At present, the only valid method available to confirm MVX infection is
based on PCR technology, by detecting the presence of viral dsRNA [2]. However, this method
is time-consuming and requires highly skilled personnel. FT-IR spectroscopy has been used
successfully in several studies to measure key parameters associated with mushroom quality
[3-5]. In this study, we investigated the use of FT-IR imaging spectroscopy as a rapid method
to effectively detect the presence of Mushroom Virus X in asymptomatic mushroom tissue

MATERIALS & METHODS

A horticultural cropping experiment was conducted at the experimental mushroom unit at
Teagasc, Kinsealy (Ireland). Control non-infected mushrooms and MVX-infected mushrooms
were grown during the course of two crops. The set of samples employed in this study was
selected using first, second and third flushes. A total of twenty-two control mushrooms and
twenty-five MVX-infected samples were used in the case of pielipellis tissue. For stalk tissue, a
smaller set was selected comprising of ten non-infected samples and nine MVX infected
samples. Following harvest, mushroom samples were subjected to a freezing pre-treatment and
stored at -80oC prior day of analysis. On the day of measurement, mushroom sections from the
pilei (cap) and stalk (stem) were cut using a Leca cryomicrotome instrument. Mushroom
sections were collected on CaF2 slides and stored at – 20oC until analysis. Spectra acquisition
took place within 8 hours of sample cutting using a Perkin Elmer Spotlight 400N over the
frequency range 700-4000 cm-1.
The microscopic images corresponding to the different mushroom tissues were sectioned
through k-means clustering analysis into homogeneous zones. The fingerprint region (830-
1750 cm-1) of the IR spectra was selected for discrimination [4]. A training subset (50% of the
total database of samples) of those spectra was used to build discrimination models of MVX
infection using Random Forest (RF) and Partial Least Squares Discriminant Analysis (PLS-
DA) classification techniques. A testing subset (the remaining 50% of the database) was used
to confirm the ability of those models to generalise this classification ability.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1773
RESULTS & DISCUSSION
The results for discrimination between MVX-infected and non-infected samples are
summarised in Table 1. The RF models resulted in good classification between MVX-infected
and non-infected control mushrooms. In particular, 98% correct discrimination of the test
subset was obtained through RF for stalk tissue. Regarding the cap tissue, the surface layer
(pileipellis) proved to be the location with the best ability to discriminate between infected and
non-infected mushrooms through changes in the FTIR spectra, with a 93% correct
classification of the test subset through RF. From the results presented in Table 1, it can be
observed that RF performance in predicting the test subset was superior compared to PLS-DA,
although both methods obtained satisfactory discrimination considering the characteristic
variability of mushroom.

Table 1. Summary of results for discrimination between MVX-Infected and non-infected control
mushrooms in the validation subset

CONCLUSION
FTIR microscopy coupled with multivariate data analysis including K-means, PLS-DA and
Random Forest offers a fast method to indicate MVX infection which can then be further
confirmed by PCR analysis

REFERENCES
[1] Grogan H.M., Gaze R.H. & Holcroft S. 2005. Viral dsRNAs in Agaricus bisporus: Transmission,
symptom expression and control. In: Tan Q., Zhang J., Chen M., Cao H., Buswell J.A (Eds.). Fifth
International Conference on Mushroom Biology and Mushroom Products. Acta Edulis Fungi, China,
363-367.
[2] Martens, H., Næs, T. 1989 Multivariate calibration.Chichester: Wiley
[3] Gaston E., et al. Prediction of Polyphenol Oxidase Activity Using Visible Near-Infrared Hyperspectral
Imaging on Mushroom (Agaricus bisporus) Caps. Journal of Agricultural and Food Chemistry,
58(10), 6226-6233.
[4] O'Gorman A., et al. Use of Fourier Transform Infrared Spectroscopy and Chemometric Data Analysis
To Evaluate Damage and Age in Mushrooms (Agaricus bisporus) Grown in Ireland. Journal of
Agricultural and Food Chemistry, 58(13), 7770-7776.
[5] Esquerre C., Gowen A.A., O'Donnell C.P., & Downey G. 2009. Initial Studies on the Quantitation of
Bruise Damage and Freshness in Mushrooms Using Visible-Near-infrared Spectroscopy. Journal of
Agricultural and Food Chemistry, 57(5), 1903-1907.

1774
 Design and validation of sensory focused processes of foods

C. Tzia, V. Giannou, D. Lebesi, D. Sabanis, V. Polychniatou, P. Sfakianakis, C. Chranioti, P. Moutsatsou

National Technical University of Athens

Laboratory of Food Chemistry and Technology
Iroon Polytechniou 9, 15780, Polytechnioupoli Zografou, Athens, Greece
Tel: +30 210 772 3165, e-mail: tzia@chemeng.ntua.gr

INTRODUCTION
Sensory characteristics are the main quality characteristics of foods that designate the consumer
choice and acceptability. The sensorial attributes are influenced during processing of foods
depending on the applied method and conditions. Sensory quality of processed foods may also be
altered during storage depending on the packaging process (packaging method, packaging material
etc.) and storage conditions. The possibility for control and maintenance of the sensory attributes of
foods is of interest for food processors. Moreover, the identification of the processes directly and
critically affecting the sensory quality of a food product is of great interest as well.
However, some food processes intend to develop one or certain sensory characteristics of food (i.e.
extraction of flavor compounds, removal of undesirable constituents, yogurt coagulation etc.). The
occurrence of a sensory attribute in respect to a certain process can be affected by various factors
(i.e. the type and the amount of a material, the proper processing conditions etc.) while the
effectiveness of such a process has important impact on the total quality of the final product. Such
sensory focused processes should be designed and validated.

MATERIALS &METHODS
Sensory attributes (color, size, form, flavor, taste and texture) of a product can be influenced by its
chemical composition and as well as by process parameters (temperature, pressure, energy input,
time, etc.), equipment (size, supplier, etc.) and storage conditions.
In the present study representative sensory focused processes of foods are presented in relation to
the sensory characteristic they induce (e.g. flavor, taste, odor, texture, appearance, color).
Thereafter the methodology for their design and validation is developed.

RESULTS & DISCUSSION

Process design, from the sensorial characteristics point of view, includes the determination of the
relative process parameters (formulation and conditions) focusing on the critical factors. A proper
methodology is proposed for design verification and process validation.
As far as the role of constituents is concerned the critical factors for the formulation is the
concentration of the ingredients (basic or minor) and their addition in the correct/optimum quantity.
Several food processes are specialized in the development of a sensory property (e.g. the critical

11th International Congress on Engineering and Food - Athens, Greece, 2011 1775
factors for the development of shape/size can be the extrusion conditions, coloring reactions
(Maillard, caramelization) or bleaching can affect color, fermentation or deodorizing induce odor
characteristics, etc.). Optimum parameters of processing and storage/maintenance can ensure the
retention of desirable properties of foods and lead to the development of intended attributes in the
final product. Heating (thermal processing), freezing, proofing, baking/grilling and aeration are
examined and the critical factors which may induce sensorial characteristics, such as time and
intensity, are recorded. Packaging and storage can also be very important. Parameters such as the
material, type and integrity of packaging may affect the formation of off-flavors or generate
oxidation. Finally, storage conditions (temperature, humidity, aeration, duration, etc.) may affect
color, odor, taste or flavor characteristics of food products through degradation, browning,
decolorization, gelling or whey off reaction. The above are summarized in Figure 1.

Figure 1. Parameters affecting the sensory properties of foods during processing.

CONCLUSION
Sensory experiments are required for process design (at different percentages of ingredients/
components used or conditions applied).
A trained sensory panel should be introduced as well as standardized methods and procedures
should be applied for sensory testing in each design/validation step.
Sensory data should be utilized for evaluation of products’ sensory quality and design verification.
Required changes should be incorporated and process should be validated.

REFERENCES
[1] Lyon D.H., Francombe M.A., Hasdell T.A. & Lawson K. 1992. Guidelines for Sensory Analysis in Food
Product Development and Quality Control, 2nd Edition. Chapman & Hall, UK.
[2] Lawless H.T. & Heymann H. 2010. Sensory Evaluation of Food: Principles and Practices, 2nd Edition.
Springer Science & Business Media, USA.
[3] Sikorski Z.E. 2002. Chemical and Functional Properties of Food Components, 2nd Edition. CRC Press
LLC, USA.

1776
 Rapid HPTLC-based method for quality control: simultaneous chemical analysis and

antioxidant activity determination in herbal, nutraceutical and functional foods

Katalina Muñoza, Jeniffer Calderóna, Edison Osorioa, Dagoberto Castrob, Raquel Sernab, Jaiber Díazb,
Julián Londoñoa

a
Universidad de Antioquia, Medellín, Colombia (kmunos@gmail.com)
b
Universidad Católica de Oriente, Rio Negro, (dcastro@uco.edu.co)

INTRODUCTION
Plant secondary metabolites, such as essential oils and flavonoids, have been widely studied for
their biological activities [1]. Nowadays, there is an increasing interest in natural antioxidants;
particularly for phenols intended to prevent not only the presumed deleterious effects of free
radicals in the human body but also the deterioration of fats and other constituents of foods.
The antioxidant property of essential oils also has been verified in vitro by physical–chemical
methods to promote their use as natural food additives [2].

Thymus vulgaris (T. vulgaris) and Rosmarinus officinalis (R. officinalis), Calendula officinalis
(C. officinalis), as well as many other aromatic plants biosynthesize high amount of volatile
compound referred as the essential oil. The whole plant and its essential oils and non-volatile
compounds can be used as natural preservative and antioxidants ingredients in the food
industries [3].

Although, antioxidants have a high demanding and also their sources, there is a great problem
for assurance raw material quality due the variability of metabolite content. This work uses
HPTLC to compare quality in terms of metabolite content and antioxidant capacity.

MATERIALS & METHODS

Plant material: For T. vulgaris (Alzate 3438) 3 materials were evaluated (T1, T2 and T3),
harvested in 6 regions in Antioquia state (Col.) Rio Negro (RN), La Ceja (LC), Marinilla (M),
Peñol (P), Guarne (G) and Santuario (S). For R. officinalis (Alzate 3444 ) 2 materials were
evaluated R1 and R2, harvested in the same locations. For C. officinalis (Alzate 3444) CI6A,
C11C, CI8K, C4, C9, C2, C2I, C9A were evaluated, cultivated in Rio Negro. Samples were
dried at 45ºC in an air-forced dryer (Dies 2009, Colombia) and ground in mill-excelsior (IKA
A11 Basic). Standars were from Chromadex.

HPTLC Analysis: Aluminium sheets Kieselgel 60 from Merck (Darmstadt, Germany) were
used. Samples were applied with a 100 uL sample syringe (Hamilton, Bonaduz, Switzerland)
using a Linomat V system (Camag, Switzerland). 5 uL were applied as 5 mm bands. Plates
were developed in a vertical glass chamber (Camag, Switzerland) for 5 min using toluene/
ethyl acetate (97:3 by volume) as a mobile phase for T. vulgaris, toluene/ethyl formiate/formic
acid for R. officinalis and ethyl acetate/formic acid/glacial acetic acid/water (100:11:11:26 by
volume) for C. officinalis. After development, the components were visualized by UV
irradiation at 280 nm for tymol, 250 nm for rosmarinic acid and 360 nm for rutin. Then the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1777
plates were dipped into a 0.05% DPPH solution. Antiradical activity was estimated on the
intensity of disappearance of violet/purple background of plate and was quantified by
densitometric scanning at 518 nm as negative peak. Stock solutions were prepared in methanol
at different concentration levels, these solutions were analyzed by HPTLC exactly as described
above and calibration curves were prepared by plotting the negative peak area versus
concentration. Antioxidant activity was expressed as ng of rutin equivalent/mg of sample.

RESULTS & DISCUSSION

Thymol, rosmarinic acid and rutin from T. vulgaris, R. officinalis and C. officinalis extracts,
respectively, were identified by comparisons of their Rf values and UV spectra to standards,
while the quantitative data were calculated from their calibration curves. The calibration curves
of negative peaks of rutin were obtained from a calibration solutions developed in the HPTLC
and postchromatographic DPPHƔ radical derivatization. A linear correlation was found between
the amount of analyte and the negative peak area (r2: 0.9735, n=4). Antioxidant activity
amongst the 18 samples of T.vulgaris was significantly different (p-value< 0.0001). For T.
vulgaris, the best material expressing the metabolite responsible of the antioxidant activity,
thymol, is T3, cultivated in S (T1S: 2.37x104 ± 4.42x102; T2S: 2.00x104 ± 1.06x103, T3S:
2.83x104 ±2.70x103 ng rutin-eqv/mg) and in Guarne (G) (T1G: 1.38x1043.58x102; T2G:
1.94x104 ±1.65x102; T3G: 2.47x104 ±4.83x101ng rutin-eqv/mg ± SD). R. officinales had a
different behavior, in this case the best places are RN (R1RN: 8.03x103±1.09x102; R2RN:
8.84x103±3.46x102) and LC (R1LC: 7.38x103 ± 6.87x101, R2LC: 1.18x104±1.95x103). The
antioxidant activity in the 12 samples tested was significantly different (p-value< 0.0001). For
C. officinalis the content of ng rutin-eqv/mg is expressed for a group of three compounds
(chlorogenic acid, hyperosid and rutin), the materials evaluated, are significantly different in
terms of their antioxidant activity (p-value< 0.0001).

CONCLUSION
In the study was observed that the effect of the ambient on the antioxidant profile is extremely
significant. The procedure can be used for rapid analysis in pharmaceutical and food industries
where standardization of raw material plays a key role on quality and efficacy of final products.
The postchromatographic derivatization with DPPH can be used as a cheap, fast, and efficient
alternative. This work pretends to demonstrate the great importance of using new technologies
in quality control.

REFERENCES
[1]. Faleiro, L., et al. 1999. Antimicrobial activity of essential oils of Rosmarinus officinalis L., Thymus
mastichina (L) L. ssp. mastichina and Thymus albicans. in II WOCMAP congress on medicinal and
aromatic plants, part 2: pharmacognosy, pharmacology, phytomedicine, toxicology.
[2]. Ruberto, G. and M.T. Baratta, 2000. Antioxidant activity of selected essential oil components in
two lipid model systems. Food chemisty. 69: p. 167-174.
[3]. Nguefack, J., et al., 2009. Food preservative potential of essential oils and fractions from
Cymbopogon citratus, Ocimum gratissimum and Thymus vulgaris against mycotoxigenic fungi.
International Journal of Food Microbiology. 131(2-3): p. 151-156.

1778
 Nondestructive evaluation of watermelon ripeness using LDV

Rouzbeh Abbaszadeha, Ali Rajabipoura, Hojjat Ahmadia, Mohammad Mahjoobb, Mojtaba Delshadc

a
Department of Mechanic of Agricultural Machinery, University of Tehran
b
Faculty of Mechanical Engineering, University of Tehran
c
Department of Horticultural Sciences, University of Tehran

INTRODUCTION
Watermelon quality during consumption, mainly depends on its extent of ripeness.In recent years
researchers have been studied a new non-destructive vibration technique using LDV technology to
test the quality of some fruits. Muramatsu , et al evaluated the texture and ripeness of some
varieties of kiwi, peach and pears. They excited samples at different stages of ripeness, by the sine
wave with frequencies from 5 to 2000 Hz and vibration at top point of the fruit were measured by
LDV. Then the phase shift between input and output signals was compared with the data obtained
from the method of force - displacement. Significant relationship between these two methods
obtained in 1200 and 1600 Hz frequencies. [1]. Terasaki , et al were used LDV to assess properties
of kiwifruit at different stages of ripeness. They consider two-factor S = fn = 22m2/3 and Ș = (f2-f1) / fn =
2 where fn = 2 : second peak resonance frequency, m: mass of fruit and f2 and f1 at 3 dB below peak
resonance are determined. The relationship between S and firmness of kiwifruit was significantly
high. Ș also showed well match with soluble solids content [2] Taniwaki , et al also conducted a
separate investigation to review the trend of change in elasticity index figures from the persimmon
and pear after harvest period. They were determined elasticity index from the formula f22m2 / 3. f2
was obtained using LDV. The samples evaluated using professional's senses and fruits ripeness were
evaluated considering features such as appearance, sweetness, firmness and etc. (each separately).
High correlation between the elasticity index and the mentioned properties were observed. [3,4].
The main goal of this research is: study the vibration response of watermelon using LDV and
developing and introducing a nondestructive method to determine the watermelon ripeness.

MATERIALS AND METHODS

In this study 14 watermelons of Crimson Sweet varietywere selected for the experiments.
First each watermelon was placed on shaker. Then samples were excited by random signal. This
signal generated by computer and were applied on a range of frequencies from zero to 300 Hz. The
signals were also amplified by a signal amplifier. For every sample, experiments were repeated in
different positions of fruit on shaker. Vibration applied to the fruit by shaker was measured by
accelerometer installed in fruit placing position and finally transmitted to the computer.
Simultaneously vibration response of fruit top point was measured by LDV Using a fast Fourier
transform algorithm with considering ratio of response signals to exciting signals, frequency
response of fruit was analyzed and the desired results was extracted.
Using frequency response curves between the accelerometer and LDV, damping ratio and resonance
frequencies of first two vibrational modes were measured. The damping ratio obtained from the
relationship Ș = (f2-f1) / f0 where Ș: the damping factor, f0:,resonance frequency, and f1 and f2 are
determined at 3 dB below the resonance peaks. phase shifts between input and output vibrations also
were considered in predetermined frequencies (50,100,150,200,250,300).In addition f02.m f02.m2 /
were also calculated using the test results applied in the analysis. After determining the vibration

11th International Congress on Engineering and Food - Athens, Greece, 2011 1779
response of the samples and measuring their weight, watermelons were sensory evaluated.
Seventeen panelists graded the fruits in a range of ripeness based on sweetness, taste (except
sweetness), color, texture and also in terms of overall acceptability (total desired traits consumers).
The fruit ripeness indices were scored on a scale of 1–5 (1: unripe, 3: ripe, and 5: overripe).Finally
the correlation between LDV-test results and the consumer opinions was determined.

RESULTS & DISCUSSION

Spectrums showed that second resonance peak (128 Hz) has better contrast and less sharpness
which can increase the accuracy.There is no significant difference between second resonance
frequency and sweetness, taste, color, texture at 1% level as well as at 5% for overall acceptability.
Index obtained f02.m2/3 has a significant correlation with color at 5% level but significant difference
was observed between quality characteristics and damping ratio and also index calculated from
f02.m at 1% level.The results shows that phase shifts at 50,100 and 300 Hz have significant
difference with sensory test results but significant relationship was observed between phase shift at
200 Hz and consumer opinions at 0.01 level. There are no significant difference between phase shift
of 150Hz and taste and overall acceptability as well as phase shift of 250 Hz and color at 0.05
level.It seems second resonance frequency and index calculated by f02.m2/3 can be used in detection
of watermelon ripeness but damping ratio and f02.m aren’t suitable for it. Meanwhile the phase shift
at 200 Hz had good relation with quality indicator. Taking into account figure 3mentioned
phenomenon can happens because of existing valley in this part of spectrum. Therefore study of
other similar points is suggested. Comparing with other methods, this technique is more accurate
without limitations and problems of acoustic method due to distribution of excitation energy in a
wide frequencies range and a period of time and lack of additional mass. Also in this method the
vibration response of watermelon using LDV is measured without direct contact, accurate and
timely that has significant advantage for grading and sorting of the melon for commercial use.

CONCLUSION
Present study demonstrates potential of laser vibrometry for predicting quality of fruit as an online
contactless sensing method. Diagnosing poor-quality watermelons in a bottleneck and separate
them, this could increase the consumer satisfaction, and providing a plan for using those products is
conceivable.

REFERENCES
[1] Muramatsu N.1; Sakurai N.; Wada N.; Yamamoto R.; Takahara T.; Ogata T.; Tanaka K.; Asakura T.; Ishikawa-
Takano Y.; Nevins D.J.(1999) Evaluation of fruit tissue texture and internal disorders by laser Doppler detection,
Postharvest Biology and Technology, Volume 15, Number 1, January 1999 , pp. 83-88(6)
[2] Terasaki, S., Wada, N., Sakurai, N., Muramatsu, N., Yamamoto, R., & Nevins, D. J. (2001). Nondestructive
measurement of kiwifruit ripeness using a laser Doppler vibrometer. Transactions of the ASAE, 44, 81–87.
[3] Taniwaki, M., Hanada, T.,Tohro, M. & Sakurai, N.(In press) Non-destructive determination of the optimum eating
ripeness of pears and their texture measurements using acoustical vibration techniques. Postharvest Biol.
Technol(2008).
[4] Taniwaki, M., Hanada, T., & Sakurai, N. (2009). Postharvest quality evaluation of‘‘Fuyu” and ‘‘Taishuu”
persimmons using a nondestructive vibrational method and an acoustic vibration technique. Postharvest Biology
and Technology. Volume 51, Issue 1, January 2009, Pages 80-85

1780
 Effect of pasteurization on bioactive amines in human milk

Fabiane Fátima Silva & Maria Beatriz A. Gloria

LBqA – Laboratório de Bioquímica de Alimentos, Faculdade de Farmácia, UFMG, Av. Antônio Carlos,
6627, CEP 31270-901, Belo Horizonte, MG, Brasil (mbeatriz@ufmg.br)

INTRODUCTION
Human milk is a complete food with the energy and nutrients requirements for the infant;
furthermore it provides protective and growth factors. Among growth factors present in human
milk, polyamines play an important role [1]. Polyamines and biogenic amines are bioactive
amines, which are aliphatic, alicyclic or heterocyclic bases. The polyamines spermine and
spermidine play important roles in growth regulation and cell proliferation, in the stabilization
of DNA, RNA transcription, protein synthesis, apoptosis and immune response regulation.
They are also relevant in the maturation of the intestinal tract. The biogenic amines are vaso-
or neuro-active. The concentration of these substances in breast milk depends on the diet and
on mothers’ conditions, such as nutrition, age, diet etc [2,3]. When mother’s own milk is not
available, processed human milk from an appropriately screened donor can be used. Human
Milk Banks (HMB) are units for the encouragement and promotion of breastfeeding as part of
public health policies. The human milk collected in HMB must be pasteurized before
distribution to infants. Pasteurization is a heat treatment (62.5 ºC/30 min) followed by rapid
cooling. It is undertaken to inactivate 100% of pathogenic microorganisms and 99.9% of
saprophyte microflora [1]. It was observed to be effect in the elimination of potential viral
contaminants such as human immunodeficiency virus, human T-lymphoma virus and
cytomegalovirus, as well as tuberculosis and other bacterial contaminants, while maintaining
the greatest possible complement of its unique bioactive factors [4]. This study was
undertaken to evaluate the influence of pasteurization on the profile and levels of bioactive
amines in human milk.

MATERIALS & METHODS

This study was conducted in a Human Milk Bank (HMB) located in Betim, metropolitan area
of Belo Horizonte, Minas Gerais, Brazil. It was approved by the Research Ethics Committee
of the Universidade Federal de Minas Gerais and of the HMB. Milk samples (n = 40) were
obtained from 21 donors. The samples were collected randomly and screened for acidity from
1 to 7 ºD. The samples of human milk were analyzed before and after pasteurization at
62.5 ºC/30 min. The profile and levels of polyamines were determined by ion-pair HPLC, post
column derivatization with o-phthalaldehyde and fluorimetric detection [2]. The data was
submitted to Anova and compared by the Mann-Whitney test at p < 0.05.

RESULTS & DISCUSSION

The polyamines spermine and spermidine were detected in the samples analyzed. The
prevalent amine was spermine, followed by spermidine in both raw and pasteurized human
milk. The levels of spermine varied from non detected (nd) to 3.29 mg/L (mean of 0.65 and
median of 0.49 mg/L) and the levels of spermidine ranged from nd to 1.31 mg/L (mean of 0.32

11th International Congress on Engineering and Food - Athens, Greece, 2011 1781
and median of 0.25 mg/L) in the raw milk. Some biogenic amines were also detected, among
them, putrescine, cadaverine, histamine, tyramine, phenylethylamine, serotonin and
tryptamine. These amines have been detected in human milk in other studies. The variation on
the levels could be related to several factors among them, health, age, number of children of
the mother. The diet of the mother can also affect the levels of amines [2,5]. As indicated in
Figure 1, pasteurization did not affect significantly the levels of the polyamines spermine and
spermidine (Mann-Whitney test, p < 0.05), which are growth and protective substances in
human milk. This result suggests that the pasteurization conditions were adequate regarding
the presence of polyamines. However, pasteurization affected significantly the levels of the
biogenic amine histamine, which increased after pasteurization. The role of histamine in
human milk must be ascertained in order to determine the impact of its changes during human
milk pasteurization.

Figure 1. Mean levels (mg/L) of polyamines and biogenic amines in human milk before and after
pasteurization at 62.5 ºC/30 min (difference for histamine, Mann-Whitney test, p < 0.05).

CONCLUSION
Pasteurization at 62.5 ºC/30 min did not affect the levels of polyamines in human milk which
are growth factors and protective substances. There was a significant increase on histamine
levels. Studies are needed to investigate the origin of this amine and its role in infant’s health.

REFERENCES
[1] Almeida S.G. & Dórea J.G. 2006. Quality control of banked milk in Brasília, Brazil. Journal of
Human Lactation, 22, 335-39.
[2] Buts J.P., Keyser N.D., Laurence R.D., Collete E. & Sokal E.M. 1995. Polyamine profiles in human
milk, infant artificial formulas, and semi-elemental diets. Journal of Pediatric Gastroenterology and
Nutrition, 21, 44-49.
[3] Larqué E., Molina M.S. & Zamora, S. 2007. Biological significance of dietary polyamines. Nutrition
23, 87-95.
[4] Braga L.P.M. & Palhares D.B. 2007. Efeito da evaporação e pasteurização na composição bioquímica
e imunológica do leite humano. Jornal de Pediatria, 83, 59-63.
[5] Dorhout B., Van Beusekom C.M., Huisman M., Kingma A.W., De Hoog E., Rudy E.B. & Muskiet
F.A.J. 1996. Estimation of 24 hour polyamine intake from mature human milk. Journal of Pediatric
and Gastroenterology Nutrition, 23, 298-302.

1782
 Integration of new/rapid methods and ICTs to improve food safety and quality

D. Lebesia, A. Bilbaob, A. I. Díazb, I. Papadakia & V. Oreopouloua

a
Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical
University of Athens, 5 Iroon Polytechniou St., 15780 Athens, Greece
b
GAIKER Centro Tecnológico, IK4 Research Alliance, Parque Tecnológico, Edificio 202, 48170
Zamudio- Bizkaia, Spain

INTRODUCTION
The control of food quality and safety is of major importance for both consumer and food
industry. Conventional analytical techniques used (e.g. chromatography, electrophoresis) do
not allow an easy continuous monitoring as they are expensive, time consuming and need well
trained operators. Consequently, new/rapid methods are continuously being developed. In the
framework of MoniQA project “Towards the harmonization of analytical methods for
monitoring food quality and safety in the food supply chain” an extensive research was
conducted to gather data about developed rapid methods of analysis and their validation status,
as well as Information and Communication Technology (ICT) systems that can be adopted by
the food industry for better management. Additionally, two surveys were carried out in
companies covering the whole food chain. The objective was to identify the current situation,
the gaps and future needs associated with the implementation of food safety and quality
management systems, and to assess implications of rapid and easy to apply methods and ICTs.

MATERIALS & METHODS

The surveys were conducted through the distribution of two questionnaires focused mainly on
the current use and future needs of rapid methods for the analyses of food hazards (specifying
the exact hazard and the rapid method used). In addition, respondents were asked to specify
which on line controllers and ICT systems they are accustomed to use routinely. More than
2600 questionnaires were circulated in 17 European and non European countries for 6 weeks to
companies covering the whole food chain e.g. raw material and ingredient suppliers, food
processing companies, retailers and catering companies in different food sectors (Bakery &
Breakfast Cereals, Dairy, Meat/Fish, Oil, Fruit/Vegetables, Prepared Foods and Beverages).

RESULTS & DISCUSSION

According to the results, several rapid methods have been developed for microbial analysis,
mycotoxins and food allergens, while few focus on the analysis of chemical contaminants
(Figure 1). Many rapid methods for microbial analysis are validated, while validation of
methods for mycotoxins and food allergens is ongoing. There are no validated methods for
several chemical contaminants (e.g. pesticides, organic environmental contaminants), while
very few rapid methods have been developed for some categories (e.g. heavy metals, food
additives, processing toxicants).The surveys among food companies indicated that the most
common type of rapid analysis used was for microbiological analytes (by 58% of the
respondents), while a smaller percentage of respondents used mycotoxin (14%) and allergen
related rapid methods (10%). However, overall the use of rapid methods in the food industries
was not that common, as indicated in Figure 1. This can be mainly attributed to

11th International Congress on Engineering and Food - Athens, Greece, 2011 1783
inadequate/poor information about existing rapid methods, except for those developed for
microbiological contaminants, and to limited existence of validated rapid methods for some
analytes, e.g. chemical contaminants and food additives. The ease of application is another
major factor, consequently, industries use mostly non validated, easy to apply test kits for food
allergens and mycotoxins.

Used Nonvalidated Validated

Pesticides
Org.environmentalcontaminants
Mycotoxins
Histamine
Heavymetals
Foodadditives
Allergens
Microbiological

0 20 40 60 80 100 120 140 160 180

Numberofrapidmethods

Figure 1. Rapid methods available and used by respondents for microbiological and chemical hazards
detection
Furthermore, companies use and need automatic on/in line monitoring systems mainly for
temperature control, as temperature is a critical parameter for the safety of the product;
automatic on/in line foreign body detection systems are also of high demand. Regarding ICT-
based management systems, there is interest in their use mainly for product and raw material
traceability and for storage area management, while ICT is also used to source information
about local legislation and regulations, existing suppliers and customers.

CONCLUSION
Rapid methods, although not used extensively, are needed throughout the food supply chain to
aid in the identification of potentially contaminated raw materials/additives/food lots.
According to the conducted survey, a lot of industries were not well informed about the rapid
methods available for evaluating the quality and safety of food and didn’t completely grasp the
essence of rapid methods, while 62% of the respondents that used rapid methods considered
that their introduction had contributed to improved food safety management. Finally, the
integration of ICTs in total quality and safety food management systems can be further
enhanced, taking into consideration the needs of food industries for improved data accuracy
and real time data acquisition.

ACKNOWLEDGMENTS
The MoniQA Network of Excellence is funded by the European Commission (contract no. FOOD-CT-
2006-36337) within the Sixth Framework Programme Topic T5.4.5.1: Quality and safety control
strategies for food (NOE). The authors appreciate the contribution of the following MoniQA partners:
Hanna-Leena Alakomi, Anton J. Alldrick, Ayse Bakan, Zsuzsanna Bugyi, Cathrine Finne Kure, Velitchka
Gotcheva, Sandra Kerbach, Susan Paulin, Xiaofang Pei, Maddalena Ragona, Christelle Robert, Kim Ahn
To and Halina Tureskja.

1784
 Commercial characterization of Madalenas: Relationship between physical and sensory

parameters

M. M. Uretaa, D. F. Oliveraa,b, V.O. Salvadoria,b

a
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), Fac. de Cs. Exactas,
UNLP – CCT La Plata, CONICET, 47 y 116, B1900AJJ, La Plata, Argentina.
b
MODIAL, Depto. Ing. Qca., Fac. de Ingeniería, UNLP, 1 y 47, B1900TAG, La Plata, Argentina.
(micaelaureta@gmail.com, danielaolivera@conicet.gov.ar, vosalvad@ing.unlp.edu.ar)

INTRODUCTION
The ability to produce high quality processed foods is essential in order to compete in a market
were consumers have exigent requests. In particular, baking industry has an additional
challenge: to achieve a mechanical and automatic process maintaining a given identity of
quality in time. Different aspects, apart from baking operative conditions, contribute to the
product quality and induce consumer’s acceptance. Among others, the change in volume and
shape, color, texture and flavor can be mentioned [1]. Madalenas, very common in Argentina
for breakfast or at tea time, are included within the general definition of sweet baked products.
They have a particular taste and shape, similar to a small cake, with a formulation that basically
consists in eggs, sugar, butter, flour and yeast.
The objective of the present work was to analyze the physical and sensory quality of
commercial madalenas with the purpose of obtaining relationships between consumer’s
preference and physical parameters.

MATERIALS & METHODS

Four different trademarks of madalenas (MC1, MC2, MC3 y MC4), normally encountered in
Argentinean food stores, were used in this study. The instrumental analysis consisted in:
texture characterization through compression [2], relaxation [2] and TPA tests using a
TA.XT2i Texture Analyzer (Stable Micro Systems, UK); water content [2] and water activity
using an AquaLab CX-2 (Decagon, USA); porosity measured by image analysis; color using a
colorimeter (Minolta CR-300, Japan); and global and crumb density. All tests were performed
by quadruplicate for each trademark. The sensory characteristics of madalenas were measured
by an untrained panel of 35 consumers (23 % males and 77 % females) using the acceptance
test, i.e. the affective method in a verbally structured 9-point hedonic scale (from 9 - extremely
like, 5 - neither like nor dislike, to 1 - extremely dislike).

RESULTS & DISCUSSION

The results of analysis are shown in Table 1. A good correlation between structural parameters
(crumb density), and textural (firmness) was found (R2!0.9). This is, the greater the density,
more firmness the sample showed.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1785
The joint analysis of these results and the previously presented physical properties showed that
acceptability are strongly correlated with porosity, crumb density and firmness (R2=0.97,
0.87and 0.92, respectively). It showed a high preference for products more porous and less firm
[3].
Although TPA is an excellent test for texture analysis in many kind of food, in this case, it
didn’t reveal significant differences between samples. On the other hand, there were found
important variation when color was evaluated, what contributed to characterize each
trademark.

Table 1. Results from all the tests performed for the analysis.
MC1 MC2 MC3 MC4
Water content (%) 17.70a 17.97a 17.91a 16.35b
Water activity 0.77a 0.77 a 0.78a 0.78a
Crumb density (g/cm3) 0.66a 0.60b 0.53c 0.73d
Global density (g/cm3) 0.50a 0.47a 0.43a 0.46a
Porosity (cell/cm2) 6.80a 4.80b 10.40c 2.30d
Firmness (N) 85.75a 69.04b 23.15c 138.97d
Elasticity (%) 48.00 a 45ab 45a 51b
Cohesiveness 0.47 a 0.51b 0.50b 0.55b
Acceptability (%) 73 % 71 % 80 % 64 %
Values followed by a different letter are significantly different (P < 0.05).

CONCLUSION
The present study is a preliminary stage of a more complete research that is focused on the
optimization of baking process conditions affecting the final quality of sweet baked products. It
showed that sensory analysis alone is not sufficient to establish commercial trends for a given
product. However, if it is accompanied by physical and texture evaluation, it can be correlated
to define the desirable quality and, what is more, it will be a useful tool to design an industrial
process with high quality standards of production.
In this case, consumers appreciate more soft and aerated product, so there is a challenge to
achieve a process that ensures these characteristics on sweet baked products.

REFERENCES
[1] van Kleef, E., van Trijp, H.C.M. and Luning, P. 2005. Consumer research in the early stages of new
product development: a critical review of methods and techniques. Food Quality and Preference, 16,
181–201.
[2] AACC. 2000. Approved Methods of the American Association of Cereal Chemists, St Paul, MN,
AACC.
[3] Morr, C.V., Hoffmann, W. and Buchheim, W. 2003. Use of applied air pressure to improve baking
properties of whey protein isolates in angel food cakes. Lebensmittel Wissenchaft und Technologie,
36, 83-90.

1786
 Integrating strain variability in modelling Salmonella enterica growth

Alexandra Lianou, Koutsoumanis Kostas

Aristotle University Of Thessaloniki

ABSTRACT
Strain variability of the growth behavior of foodborne bacterial pathogens has been well
documented and acknowledged as an issue of major importance in food safety management.
Aiming at integrating such a variability in predictive modelling approaches, a model
incorporating intra-species variability data and predicting the maximum specific growth rate
(ȝmax) of Salmonella enterica as a function of pH and aw was developed.
For this purpose, growth kinetic data (ȝmax values) of 60 S. enterica isolates, generated during
monitoring of growth at 37°C in tryptone soy broth of different pH (4.0-7.0) or aw values
(0.964-0.992), were used. The effects of pH and aw on ȝmax were modelled for each tested
strain using a cardinal parameter model (CPM), and the parameters pHmin, pHopt, awmin, awopt
were estimated and used to describe the variability of ȝmax among the strains. Specifically, S.
enterica strain variability was described by introducing cumulative frequency distributions for
the cardinal parameter values into the model. In this way, the prediction of ȝmax provided by the
model using Monte Carlo simulation is a distribution of values, allowing for the strain
variability of this growth kinetic parameter to be taken into account. The developed model was
validated against growth kinetic data generated for the 60 strains of the pathogen at pH 5.0-aw
0.977.
The model predicted accurately the ȝmax of S. enterica, with the mean and standard deviation of
the latter being 0.78 and 0.09 h-1, respectively, and with 5th and 95th percentiles of 0.62 and
0.91 h-1, respectively. The mean, standard deviation, and the 5th and 95th percentiles of the
observed ȝmax values were 0.73, 0.09, and 0.61 and 0.85 h-1, respectively.
The stochastic approach developed in this study can be useful in describing and incorporating
the strain variability of S. enterica growth kinetic behavior in predictive microbiology and
microbial risk assessment.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1787
 A study on germination time and mycelium growth kinetics of single fungal spores

Maria Gougouli, Kostas Koutsoumanis

Aristotle University Of Thessaloniki

ABSTRACT
Most of the available studies on fungal behaviour in foods deal with monitoring growth of
mycelia originated from a large number of spores. In practice however, the contamination of
food with fungal spores occurs at very low numbers. Therefore, the study of large populations
can lead to unreliable conclusions, because it does not take into account the variability between
individual spores.
This study aims at describing the germination time and mycelium growth kinetics of single
fungal spores and establishing a relationship between them.
The germination time and the mycelium growth kinetics of single fungal spores of Penicillium
expansum and Aspergillus niger were monitored at isothermal conditions ranging from 0 to 33
°C and from 5 to 42 °C, respectively. For each temperature, the germination time of 450 spores
on malt extract agar was monitored microscopically, while the radial growth of 200 mycelia
originated from a single spore was assessed macroscopically. The data on the germination and
the lag time of single spores were expressed as cumulative frequencies and fitted to the logistic
model. In addition, a time-lapse microscopy method was developed for monitoring single spore
behaviour using a z-motorized microscope (Olympus BX61) equipped with ×40 and x4
objectives (Olympus) and a high-resolution device camera (Olympus DP71). The quality of the
images was improved by developing an auto focus procedure with an Extended Depth of Focus
(EDF) system using the ScopePro module of ImageProPlus software. The output of the above
method was a video for each spore showing the behaviour of single spores from germination to
mycelium formation.
The results of the study showed similar temperature dependence for the germination and the
lag time of single spores. However, the temperatures limits for germination time were wider
compared to the lag time due to the fact that at the growth boundary spores germinated but
were not able to form a mycelium. The lag time was significantly longer that the germination
time. This was confirmed by the spore behaviour videos which clearly showed the relation
between spore germination time and the lag calculated from the mycelium growth.
The data provided by this work describe the relation between germination and lag time of
single spores and their variability, and can help to the development of stochastic methods for
determining the risk of food spoilage by fungi.

1788
Quantifying the combined effect of salt and temperature on the growth of Listeria strains

isolated from salmon and salmon processing environments

Torstein Skåra1,2, Astrid M Cappuyns2, Eva Van Derlinden2, Jan Thomas Rosnes1, Vasilis P Valdramidis3, Jan FM Van
Impe2

1
Nofima, Stavanger, Norway (Torstein.Skara@nofima.no)
2
Katholieke Universiteit Leuven, Department of Chemical Engineering, BioTeC - Chemical and Biochemical Process
Technology and Control, Leuven, Belgium (jan.vanimpe@cit.kuleuven.be)
CPMF² - Flemish Cluster Predictive Microbiology in Foods – http://www.cpmf2.be/
3
Biosystems Engineering UCD, School of Agriculture, Food Science and Veterinary Medicine University College
Dublin, Ireland (Vasilis.Valdramidis@ucd.ie)

INTRODUCTION
Recent legislation for shelf life determination of RTE-products recommends documentation of growth
potential of L. monocytogenes to be obtained either through challenge studies, or modelling [1].
Monitoring bacterial growth response in nutrient media through measurements of optical density is an
often used method for estimation of growth parameters [2]. Although validated growth models exist [3],
the slow growth response of Listeria at refrigerated storage temperatures makes growth parameter
estimation useful, for example, for screening strains and new preservation conditions. The objective of
this work is to study the effect of temperature and salt concentration on the growth of Listeria in a liquid
medium model system. The strains investigated originate from or are relevant for salmon processing.

MATERIALS & METHODS

L. monocytogenes strains (5, 11, 14, 15, 21, 26, 44, 51-2) isolated from salmon and salmon processing
environment and two L. innocua strains, ATCC 33090 and CCUG 35613 (= ATCC 51742) were
investigated with respect to their growth kinetic properties.10-fold serial dilutions of inocula grown to
stationary phase were prepared in TSBYE with 0, 2.5 and 5 % (w/v) NaCl. 5 ten-fold dilutions of each
strain were transferred to microtiter plates and mounted in a Bioscreen C measuring absorbance at 600
nm (abs600nm) at regular time intervals, at 4, 12 and 30ºC. The maximum specific growth rate, ȝmax, and
the lag time, Ȝ, were estimated from abs600nm by determination of the time to detection, and by using the
serial dilution method [4].

RESULTS & DISCUSSION

For the different salt and temperature combinations under study, no significant differences in ȝmax and Ȝ
were observed for the different strains within one species. Therefore, no individual strain results are
presented, but the results are grouped per species. The estimated ȝmax and the estimated Ȝ are presented in
table 1, showing the decrease maximum growth rate with decreasing temperature. The effect of increasing
the salt concentration from 0% to 5% led to a reduced growth rate. Overall L. innocua showed a higher
growth rate than L. monocytogenes at 30ºC, while at 4ºC L. monocytogenes showed the higher growth
rates. For L. monocytogenes the lag phase increased with decreasing temperature. Salt concentration also
affected the lag phase of L. monocytogenes. At 30°C, however, the estimated lag phase values were in the
same range for the different salt concentrations. For L. innocua an increase in lag phase can also be seen

11th International Congress on Engineering and Food - Athens, Greece, 2011 1789
for decreasing temperature. An increase of Ȝ was seen with increasing salt concentration. In general the
lag phase of L. innocua increased more than that of L. moncytogenes with increasing salt concentration
and decreasing temperature.

Table 1: Effect of temperature and salt concentration on the maximum growth rate (mean maximum growth rate, ȝmax
(h-1) ± SD) and lag phase (mean lag phase, Ȝ (h) ± SD) of L. monocytogenes and L. innocua strains in TSBYE
Estimated Temperature (ºC)
Salt (%) Species
Parameter 30 12 4
L. monocytogenes ± 1.03 ± 0.01 0.185 ± 0.004 0.042 ± 0.002
0
L. innocua ± 1.05 ± 0.02 0.179 ± 0.003 0.034 ± 0.001
ȝmax L. monocytogenes ± 0.85 ± 0.02 0.158 ± 0.006 0.033 ± 0.003
2.5
L. innocua ±± 0.89 ± 0.03 0.157 ± 0.001 0.023 ± 0.002
L. monocytogenes ± 0.49 ± 0.05 0.124 ± 0.007 0.028 ± 0.002
5
L. innocua ±± 0.62 ± 0.02 0.119 ± 0.007
L. monocytogenes 1.1 ± 0.1 5.7 ± 0.7 37 ± 7
0
L. innocua 1.5 ± 0.5 6.8 ± 1.7 80 ± 8
L. monocytogenes 1.3 ± 0.2 6.5 ± 1.4 23 ± 8
Ȝ 2.5
L. innocua 1.7 ± 0.3 10.0 ± 0.7 41 ± 20
L. monocytogenes 1.0. ± 0.5 9.9 ± 1.6 53 ± 9
5
L. innocua 2.2 ± 0.5 15.5 ± 2.4

CONCLUSIONS
The results show an increase in growth rate of Listeria with increasing temperatures. The levels were
quite similar for both species, although L. monocytogenes showed the higher growth rate at 4ºC. Lag
phase was more affected than growth rate; both by temperature and salt concentration, and these effects
were most prominent for L. innocua. As the major factors for controlling growth of Listeria in salted fish
products, the effects of temperature and salt content are relevant to product development and food safety.

ACKNOWLEDGEMENTS: This work was funded by The Research Council of Norway (project no.
186905). The authors wish to thank the National Institute of Nutrition and Seafood Research
(www.nifes.no) for providing the strains and Stord/Haugesund University College for lending their
Bioscreen C instrument. E. Van Derlinden is supported by grant PDMK/10/122 of the Research Fund of
the Katholieke Universiteit Leuven. Jan Van Impe holds the chair Safety Engineering sponsored by the
Belgian chemistry and life sciences federation essenscia. This work was supported by the Belgian
Program on Interuniversity Poles of Attraction, initiated by the Belgian Federal Science Policy Office.

REFERENCES
[1] COMMISSION REGULATION (EC) No 1441/2007 of December 2007 amending Regulation (EC) No 2073/2005
on microbiological criteria for foodstuffs. L 322, 12-29. 2007. Official Journal of the European Union.
[2] Augustin J.C., Rosso L., & Carlier V. 1999. Estimation of temperature dependent growth rate and lag time of
Listeria monocytogenes by optical density measurements. Journal of Microbiological Methods, 38(1-2), 137-
146.
[3] Mejlholm O. & Dalgaard P. 2007. Modeling and predicting the growth boundary of Listeria monocytogenes in
lightly preserved seafood. Journal of Food Protection, 70(1), 70-84.
[4] Biesta-Peters E.G., Reij M.W., Joosten H., Gorris L.G.M., & Zwietering M.H. 2010. Comparison of two optical-
density-based methods and a plate count method for estimation of growth parameters of Bacillus cereus.
Applied and Environmental Microbiology, 76(5), 1399-1405.

1790
 Modelling thermosonication inactivation of Aspergillus flavus combining natural

antimicrobial at different pH

Claudia P. Coronela, Maria T. Jiméneza, Aurelio López-Maloa, Enrique Paloua

a
Universidad de las Américas Puebla, Cholula, Puebla, México (claudiap.coronelaa@udlap.mx)

INTRODUCTION
High-intensity ultrasound represents an alternative for the development of minimally processed
foods as a possible emerging preservation factor in combination with other hurdles to reach the
desired inactivation effect [1]. The introduction of any alternative technology or combination
of various alternative/traditional technologies requires scientific data about microbial response.
Kinetic parameters and models are essential to develop food preservation processes that ensure
safety. The parameters also allow comparison of the ability of different process technologies to
reduce microbial populations. An alternative survival model is the Weibull distribution
function that presents the main advantage of remaining very simple and being sufficiently
robust to describe both downward concave survival curves (n > 1) and upward concave curves
(n < 1), and the model includes the traditional case where the survival curve is linear [2, 3].
The objective of this study was to model the combined effect of thermosonication at different
pH on the inactivation kinetics of A. flavus.

MATERIALS & METHODS

A. flavus was inoculated into sterile broth prepared with Sabouraud glucose 2% broth adjusted
by sucrose addition to aw 0.99 and citric acid to pH 5.5 or 3.0 and 500 ppm vanillin added, then
treated with combining thermosonication (52.5, 55 and 57.5°C and ultrasound at 20 kHz with
13 mm diameter probe operating continuously at an amplitude of 60, 90 and 120 μm). Survival
viable mold spore counts were determined immediately after treatment by surface plating on
potato-dextrose agar and incubated at 27°C for 3-5 days. Survival data were fitted to the
cumulative form of frequency Weibull distribution of resistances.

RESULTS & DISCUSSION

Weibull model successfully captured all dose-response curves with a satisfactory fit and lower
mean square error values (MSE). Predicted survival curve corresponding to 57.5°C, amplitude
120μm and pH 3.0 followed a first order kinetics. Shape factors (n) in Table 1 indicate that
most of the tested thermoultrasonic treatments were concave downward (n > 1), while few
treatments were concave upward (n < 1). A. flavus exhibited higher sensitivity at lower pH 3.0
and it is noticeable that at pH 5.5, in most of the cases, n was higher than 1, which suggests
that the remaining cells become weaker when treatment time increases, indicating that
accumulated damage may occur due to combined stress factors. In this study, increasing the
amplitude at pH 3.0 changed the survival patterns of the mold, however at higher pH 5.5 the
change of ultrasound intensity (higher amplitude) had no influence on the survival patterns.
Treatment at 90 μm amplitude frequency distribution parameters (mode, mean, variance and
coefficient of skewness) indicated overall spread of data with corresponding mean values
greater than for 60 μm and 120 μm amplitudes at 57.5°C at pH 3.0 or 5.5.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1791
 Table 1. Weibull distribution parameters (b, n) for A. flavus under thermosonication treatments at
selected pHs and 500 ppm vanillin.
pH T Ampli- b n R2 MSE Mode Mean Varian- Coefficien
(°C) tude (min) (min) ce t of skew-
(μm) (min2) ness (-)
3.0 52.5 60 0.071 1.224 0.985 0.012 2.175 8.158 44.896 0.066
90 0.127 0.942 0.952 0.004 -- 9.150 94.443 0.020
120 0.227 0.886 0.962 0.009 -- 5.650 40.824 0.038
55.0 60 0.102 1.220 0.977 0.003 1.595 6.099 25.271 0.091
90 0.040 1.827 0.941 0.008 3.753 5.147 8.525 0.620
120 0.270 0.814 0.988 0.001 -- 5.595 47.915 0.032
57.5 60 0.727 0.733 0.983 0.005 -- 1.874 6.771 0.210
90 0.402 1.186 0.875 0.027 0.452 2.032 2.957 0.317
120 0.543 1.031 0.975 0.008 0.060 1.787 3.007 0.307
5.5 52.5 60 0.280 0.696 0.953 0.007 -- 7.936 136.328 0.012
90 0.035 1.930 0.971 0.002 3.895 5.043 7.406 0.828
120 0.292 0.678 0.976 0.002 -- 8.019 147.949 0.012
55.0 60 0.074 1.542 0.939 0.009 2.735 4.851 10.316 0.291
90 0.071 1.539 0.920 0.020 2.829 5.039 11.171 0.281
120 0.041 1.737 0.940 0.017 3.833 5.596 11.045 0.467
57.5 60 0.379 1.473 0.981 0.005 0.897 1.748 1.455 0.541
90 0.136 2.390 0.960 0.017 1.837 2.043 0.828 1.679
120 0.510 1.304 0.981 0.007 0.549 1.547 1.431 0.500

In addition, the lower amplitude (60 μm) curve was more skewed to the right than curves for
the other amplitudes evaluated, showing that an important fraction of the mold can survive
under this treatment. The higher distribution mean values, had much more spread evidenced by
the high variance values (Table 1) at the lowest temperature evaluated (52.5°C).

CONCLUSION
Weibull parameters are useful to explain mold inactivation and perform comparisons among
treatments. The pH treatment medium had an important effect for inactivating the mold since
much lower frequency distribution of resistances were observed at lower pHs. Combining
thermoultrasonication, vanillin and pH allows shorter treatment times for mold inactivation

REFERENCES
[1] Guerrero S., López-Malo A. & Alzamora, S.M. 2001. Effect of ultrasound on the survival of Saccharomyces
cerevisiae: influence of temperature, pH and amplitude. Innovative Food Science and Emerging Technologies,
2(1), 31-39. [2] Alzamora S.M., Guerrero S., López-Malo A., Palou E., Char C.D. & Raffellini S. 2009. Models
for microorganism inactivation: application in food preservation design. In “Processing Effects on Safety and
Quality of Foods”. E. Ortega (ed.). CRC Taylor and Francis, New York, USA. [3] Mafart, P., Couvert, O.,
Gaillard, S. and Leguerinel, I. 2002. On calculating sterility in thermal preservation methods: application of the
Weibull frequency distribution model. International Journal of Food Microbiology 72, 107-113.

1792
 Survival of Bifidobacterium longum in model solutions and fruit juices

Sawaminee Nualkaekul1, Ivan Salmeron2, Dimitris Charalampopoulos1

1
Department of Food and Nutritional Sciences, University of Reading, PO Box 226, Reading RG6 6AP, UK
2
Facultad de Ciencias Quimicas, Universidad Autonoma de Chihuahua, Chihuahua, Chih. Mexico C.P. 31125

INTRODUCTION

Although significantly less amount of research has been conducted on the factors that influence the viability of
probiotics during storage in fruit juices compared to dairy products, it has been shown that the storage temperature and
time [1] and the presence of dietary fibre in the product [2] are likely to have an important role. However, there is very
little information on the effect of the chemical composition of the fruit juices on probiotic survival. The aim of the
study was to develop a mathematical model describing the survival of a model probiotic B. longum strain during
refrigerated storage in model solutions, as a factor of the pH, and the citric acid, protein and dietary fibre
concentrations. These results were then used to explain the survival of B. longum in various types of fruit juices taking
into account their composition.

MATERIALS & METHODS

Preparation of bacterial culture
Bifidobacterium longum NCIMB 8809 (National Collection of Industrial and Marine Bacteria, UK) was used in this
study. The cells were grown in trypticase-phytone-yeast extract medium at 37 ºC for 16 h.
Experimental design
Four factors (pH, citric acid, protein and dietary fibre) were studied at three different levels; pH 3.2-4, citric acid 2-15
g/l, protein 0-10g/l, dietary fibre 0-8 g/l. The experiments were designed on the basis of a Central Composite Design
(CCD). In total, thirty-one experimental runs were performed. Multiple regression analysis was carried out in order to
fit a second order polynomial equation describing the log decrease [log10N0week-log10N6 week], as a function of the four
factors. The basal medium for all solutions contained 50 g/l sucrose, 25 g/l glucose and 25 g/l fructose. The initial cell
concentration was approximately 1× 108 CFU/ml. The solutions were stored at 4 °C for 6 weeks. Samples were
collected every week for analysis.
Chemical analyses
Sugar concentrations (sucrose, glucose and fructose) and organic acids were determined by high performance liquid
chromatography (HPLC). The total dietary fibre in the juices was determined by a combination of enzymatic and
gravimetric methods (AOAC, 1997), whereas the total phenol by the Folin–Ciocalteu assay.
Cell survival in fruit juices
Six commercial fruit juices (orange, grapefruit, blackcurrant, pineapple, pomegranate and strawberry juice) were
purchased from a local supermarket. The PBS/cell suspension was added into 40 ml of each juice, so that the initial cell
concentration was approximately 1 × 108 CFU/ml, and the juices stored at 4 °C for 6 weeks. Samples were collected
weekly and analysed for pH, cell concentration and composition.

RESULTS & DISCUSSION

ANOVA analysis of the stepwise regression model demonstrated that a second order polynomial model fitted well the
data. Based on the regression coefficient estimates, it was deduced that all the controlling factors, i.e., pH, and citric
acid, protein and dietary fibre concentrations had a significant effect on the model. The resulting model describing the
log decrease [log10N0week-log10N6week] as a function of the four factors was:
[log10N0week- log10N6week] = 4.051-0.703[pH]-0.152[citric acid]-0.151[protein]+0.005[citric acid][citric
acid]+0.009[dietary fibre][dietary fibre]+0.026[pH] [protein]-0.026[pH][dietary fibre]+0.003[citric acid][protein]

11th International Congress on Engineering and Food - Athens, Greece, 2011 1793
Figure 1 shows the contour plots of the log decrease [log10N0week- log10N6week] as a function of two factors, with the
other being kept constant at the middle point values. Taking into account the plots, it can be derived that good survival
of B. longum (log decrease < 0.4 log) was obtained in the model solutions were the pH was above 3.7, the citric acid
concentration 8-15 g/l, the protein concentration above 3 g/L and the dietary fibre concentration 3-7 g/l.

Figure 1 Contour plots describing the dependence of the

log decrease [log10N0week- log10N6week] as a function of
two controlling factors. A): pH and citric acid (CA),
protein 5 g/l, dietary fibre (DF) 4g/l ; B): pH and
protein, CA 8.5 g/l, DF 4 g/l; C): pH and DA, CA 8.5 g/l,
protein 5 g/l; D): CA and protein, pH 3.6, DF 4 g/l.

It has been well established that the pH is an important

factor influencing the survival of probiotic bacteria in
food products. Regarding the role of citric acid, up to
now, there has been no information in the literature on
the role of organic acids in the survival of lactic acid
bacteria in food products during storage. It was
expected that organic acids would have a negative
effect. However, according to the results from the
model solutions, citric acid was seen to have a significant (P < 0.05) positive effect, especially when present at
concentrations between 8 g/l and 15 g/l (Figure 1). Possible explanations for this could be that citric acid was either
metabolised by the cells during storage, although HPLC analysis did not show any significant (P < 0.05) decreases
(data not shown), or that there was some kind of buffering effect by citric inside the cells.
The model predicted well the survival of the Bifidobacterium strain in grapefruit and pineapple and adequately in
orange and blackcurrant. However, it failed to predict the cell survival in pomegranate and strawberry. This suggests
that although the four controlling factors that were studied in this work, namely pH, citric acid, protein and dietary
fibre, are important factors influencing the cell survival in fruit juices, additional ones, for example the levels of
phenols in the juices, are likely to be involved. The highest cell survival (less than 0.4 log decrease) after 6 weeks of
storage was observed in orange and pineapple, both of which had a pH of about 3.8. Although the pH of grapefruit and
blackcurrant was similar (pH ~ 3.2), the log decrease of the former was ~ 0.5 log, whereas of the latter was ~ 0.7 log.
One reason for this could be the fact that grapefruit contained a high amount of citric acid (15.3 g/l). The log decrease
in pomegranate and strawberry juices was extremely high (~8 logs) most likely due to the high levels of total phenol.

CONCLUSIONS

Based on the storage experiments of Bifidobacterium longum in model solutions and the mathematical model that was
developed it was shown that high levels of pH, citric acid, protein and dietary fibre enhanced the cell survival during
refrigerated storage. The mathematical model was able to predict adequately cell survival in orange, grapefruit,
blackcurrant and pineapple juices. However, the model failed to predict cell survival in pomegranate and strawberry, in
which the cell viability declined rapidly, most likely due to the very high levels of phenolic compounds in these two
juices.

REFERENCES

[1] Saarela, M., Alakomi, H.L., Puhakka, A., & Matto, J. 2009. Effect of the fermentation pH on the storage stability
of Lactobacillus rhamnosus preparations and suitability of in vitro analyses of cell physiological functions to predict it.
Journal of Applied Microbiology, 106, 1204-1212.
[2] Saarela, M., Virkajarvi, I., Alakomi, H.L., Mattila, & P.S., Matto, J. 2006. Stability and functionality of freeze-
dried probiotic Bifidobacterium cells during storage in juice and milk. International Dairy Journal, 16, 1477–1482.

1794
 Inactivation kinetics of attached Escherichia coli cells on stainless steel and fresh-cut

apples by hydrogen peroxide disinfection treatments

Raffellini, S.a, Ortiz, S.a, Guerrero S.N.b, Alzamora, S.M.b

a
Department of Technology, University of Luján, 6700 Luján, Argentina.(sraffellini@gmail.com)
b
Natural and Exact Sciences School, Buenos Aires University, Ciudad Universitaria, 1428, Ciudad
Autónoma de Buenos Aires, Argentina (smalzamora@gmail.com)

INTRODUCTION
The foodborne illness outbreaks linked to fresh-cut fruits and vegetables have increased in the
last years. Chemical disinfection is one of the most critical processing steps in fresh-cut
vegetable production [1]. Most of the literature regarding the use of sanitizers concluded that
the disinfection process reduces the microbial populations on the surface of the produce by
only 2 log units. This behaviour is possibly due to the attached microbial cells are more
resistant to sanitizer treatments than the corresponding planktonic cultures. Hydrogen peroxide
(H2O2) has been proposed as an alternative for decontaminating fruit and vegetables. However,
there is lack of quantitative information concerning its efficacy for inactivating attached
microbial cells on different surfaces. The objective of this study was to evaluate and to model
the survival of attached E. coli cells on stainless steel and fresh-cut apple slices as affected by
H2O2 treatment at different concentrations and temperatures.

MATERIALS & METHODS

Stainless steel coupons and fresh-cut apple slices (10 x 50 mm) were suspended in 12 mL
Tryptic Soy Broth and the growth medium was inoculated with E. coli ATCC 35218. Test
samples were harvested after 48 h at 37ºC, when they reached §106 E. coli cells/cm2 (adhesion
by immersion). Additionally, apple slices were inoculated by spreading with E. coli ATCC
35218 and incubated in sterile Petri plates 24 h at 37ºC. Samples with attached bacterial cells
were rinsed in phosphate buffer and suspended in sterile citric acid – Na2HPO4 buffer solutions
(pH 3.0) containing H2O2 solutions (0; 2 and 3% w/v) at 25ºC and 50ºC. The treated samples
were periodically collected, neutralized by immersion in 4% w/v Na2S2O3.5H2O solution, and
shaken 2 min to dislodge attached cells. Dislodged cells were serially diluted in 0.1% peptone
water, pour plated using Plate Count Agar and counted after incubation at 37° C for 48 h.

RESULTS & DISCUSSION

The semilogarithmic experimental survival curves of stainless steel attached E. coli cells
treated with different H2O2 concentrations and temperatures are presented in Figure 1. For
comparative purposes, Figure 1 includes survival curves of planktonic cells obtained in a
previous research [2]. H2O2 efficacy was less as concentration and temperature decreased. At
50ºC, 3% w/v H2O2 caused 5.0 log reductions of attached cells after 1 min exposure while 4
min were necessary to obtain the same reduction when using 2% w/v H2O2. The same
treatment caused 5.0 log reductions in planktonic E. coli cells with exposures of less than 1

11th International Congress on Engineering and Food - Athens, Greece, 2011 1795
minute [2]; therefore attached E. coli cells were more resistant to H2O2 treatments than
corresponding planktonic cultures.
Time (min)
0 5 10 15 20 25

-0.5

¯ Control
Ŷ  2.0% H2O2 – 25°C – attached cells
-1.5
Ÿ  3.0 % H2O2 – 25°C – attached cells
Ɣ  ˜  2.0% H2O2 – 25°C – planktonic cells
-2.5
¡ 3.0% H2O2 – 25°C – planktonic cells
Log (N/N0)

Ƒ--- 2.0% H2O2 – 50°C – attached cells

ǻ 3.0% H2O2 – 50°C – attached cells
-3.5

-4.5

-5.5

-6.5

Figure 1. Effect of H2O2 concentration (% w/v) and temperature on semilogarithmic survival curves of
planktonic and stainless steel attached E. coli ATCC 35218 cells. Experimental (points) and fitted
values derived from the Weibullian model (lines). Data of planktonic cells obtained from [2].

Cells attached on apple slices were even more resistant, and different mechanisms of adhesion
to the tissue caused different resistance to H2O2: 3.6 and 10.5 min were necessary to obtain 5-
log reductions in attached cells by immersion or spreading respectively with 3.0% w/v H2O2 at
50°C. Survival curves of attached cells on stainless steel and fresh-cut apples were not linear
unlike those reported for planktonic cultures for the same H2O2 treatments [2]. Inactivation
kinetics of attached cells on stainless steel with enough experimental data was modelled by
using a Weibull type distribution of resistances model (Figure 1). Experimental curves were
highly correlated to fitted data, obtaining significant adjusted coefficients of determination
R2adj (0.96–0.99). Heavy tails in curves derived from the model would be indicating the
presence of attached cell subpopulations highly resistant to H2O2 treatments [3].

CONCLUSION
This study confirmed that model systems of attached cells to inert or fruit surfaces provide
results that are more comparable to the actual situations in food processing than those
performed with planktonic cells.

REFERENCES
[1] Gil M., Selma M., López-Gálvez F. & Allende A. 2009. Fresh-cut product sanitation and wash water
disinfection: problems and solutions. International Journal of Food Microbiology, 134(1-2), 37-45.
[2] Raffellini S., Schenk M., Guerrero S. & Alzamora S.M. 2011. Kinetics of Escherichia coli
inactivation employing hydrogen peroxide at varying temperatures, pH and concentrations. Food
Control, 22(6), 920-932.
[3] van Boekel M.A.J.S. 2002. On the use of the Weibull model to describe thermal inactivation of
microbial vegetative cells. International Journal of Food Microbiology, 74(1-2), 139–159.

1796
 Bi-phasic growth of Listeria monocytogenes Scott A in Modified Welshimer’s broth at 7,

10 and 14°C

Nikolaos A. Tyrovouzisa, Apostolos S. Angelidisb and Nikolaos G. Stoforosc

a
Aristotle University of Thessaloniki, Department of Chemical Engineering, Greece (tyrovou@auth.gr)
b
Aristotle University of Thessaloniki, School of Veterinary Medicine, Greece (asangel@vet.auth.gr)
c
Agricultural University of Athens, Department of Food Science and Technology, Greece
(stoforos@aua.gr)

INTRODUCTION
The psychrotrophic nature of Listeria monocytogenes is one of the most salient characteristics of
this food-borne pathogen and highly important for food processors. Research has shown that the
pathogen’s ability to proliferate at low temperatures, involves, amongst others, the intracellular
accumulation of compatible solutes such as glycine betaine and L-carnitine, which act as
cryoprotectants [1]. Defined media are used in microbiology in order to conduct reproducible
experiments and avoid confounding by extraneous, often-unknown factors originating from the
composition of the growth medium. Modified Welshimer’s Broth (MWB) is the defined medium
that has been most frequently used by researchers working on the physiology and genetics of
L. monocytogenes [2].
The objective of this work was to further explore a previous observation [3] concerning the
bi-phasic growth of L. monocytogenes Scott A in MWB at 7°C. In the current work, further
experiments were conducted with L. monocytogenes Scott A in MWB at 4°C, 7°C, 10°C, 14°C and
18°C. At each tested temperature, cultures were grown both in the presence (20 ȝȂ) or absence of
L-carnitine. Cell density was directly measured by surface-plating aliquots from each culture at
selected time intervals. Additionally, in four sets of experiments at 7°C, the composition of MWB
was modified using a rotation scheme of ten-fold increases in different sets of medium constituents.

MATERIALS & METHODS

L. monocytogenes cultures were grown overnight at 30°C in Brain Heart Infusion (BHI) broth, and
1-mL aliquots were centrifuged at 10000 rpm for 10 min. The pellets were washed twice with 1-mL
of MWB and used to inoculate (1%) 125-mL flasks containing 50 mL MWB. The cultures were
grown with mild shaking (120 rpm) at 30°C for 48 hours, until stationary phase to cell densities of
ca. 109 CFU/mL. Stationary phase cultures were then serially diluted in MWB and used to inoculate
10 sets of 125-mL flasks containing 50 mL of MWB. These cultures were incubated aerobically
(120 rpm) at 4°C, 7°C, 10°C, 14°C and 18°C with or without 20 ȝM L-carnitine. Growth was
monitored by spreading appropriately diluted aliquots in triplicate on the surface of BHI plates.
Plates were incubated at 30°C for 48 h and the CFU/mL was determined for each sampling point
based on viable counts. The different trials were tested at least in duplicate.

RESULTS & DISCUSSION

Experimental data for L. monocytogenes growth in MWB are presented on Figure 1. The growth of
L. monocytogenes in MWB at 7°C and 10°C (but not at 4°C or 18°C) was bi-phasic irrespective of

11th International Congress on Engineering and Food - Athens, Greece, 2011 1797
the presence of L-carnitine. At 14°C, growth was bi-phasic in the absence of L-carnitine, but not
when the cryoprotectant was added in the growth medium. At 4°C, growth was observed only in the
presence of L-carnitine. The effect of temperature and L-carnitine addition was assessed by
estimating, through Gompertz equation, the kinetic parameters of L. monocytogenes growth.
In order to investigate whether the observed growth phenotypes have a nutritional basis, four
modifications in the composition of the growth medium (MWB) were tested at 7°C. These data
indicated either that the observed bi-phasic growth in MWB, at least at 7°C, does not have a
nutritional basis, or that a different combination of MWB constituents, other than the combinations
tested in this work, needs to be augmented in order to alleviate growth.
10

8
log(CFU/mL).

4
4°C 7°C 10°C
14°C 18°C
2
0 10 20 30 40 50 60
Time (days)

Figure 1. Growth of L. monocytogenes in MWB at different temperatures. (Lines represent Gompertz equation
fitting to the experimental data points.)

CONCLUSION
The shape of the growth curves for L. monocytogenes cultures in MWB was function of the growth
temperature, and to a lesser degree, of the presence of L-carnitine in the growth medium. It
appeared that bi-phasic growth occurred when experimental conditions allowed for intermediate
growth rates. Modifications in the concentration of MWB constituents did not appear to influence
the bi-phasic nature of L. monocytogenes growth when tested at 7°C.

REFERENCES
[1] Angelidis A.S. & Smith G.M. 2003. Role of the glycine betaine and carnitine transporters in adaptation of
Listeria monocytogenes to chill stress in defined medium. Applied and Environmental Microbiology,
69(12), 7492-7498.
[2] Premaratne R.J., Lin W.-J. & Johnson E.A. 1991. Development of an improved chemically defined
minimal medium for Listeria monocytogenes. Applied and Environmental Microbiology, 57(10), 3046-
3048.
[3] Tyrovouzis N.A., Angelidis A.S., Liakopoulou-Kyriakides M. & Stoforos N.G. 2008. Effect of
cryoprotectants on the growth characteristics of Listeria monocytogenes at low temperatures. 10th
International Congress on Engineering and Food, Viña del Mar, Chile. Paper No. K-04.

1798
 Kinetic of White Chocolate Color Loss

Denise C. P. JARDIMa; Aline G. ORSEb; Priscilla EFRAIMa; Silvia C. S. R. de MOURAa

a
Instituto de Tecnologia de Alimentos (ITAL), Campinas, Brazil (djardim@ital.sp.gov.br,
efraim@ital.sp.gov.br, smoura@ital.sp.gov.br)
b
Universidade Metodista de Piracicaba (UNIMEP), S. Barbara d´Oeste, Brazil (alineorse@gmail.com).

INTRODUCTION
Quality loss caused by yellowing (loss of white color, darkening or browning) during its
storage and marketing is considered the main reaction which limits the shelf-life of white
chocolate [1]. The system sugar, fat and milk is susceptible to browning reactions, mainly by
Maillard reaction, especially because of the sugars and amino acids in the composition. The
occurrence of reaction depends on the conditions of humidity and temperature, as well as on
changes to these conditions, during chocolate storage. The kinetics data (constant and order of
reaction, activation energy (Ea) and factor Q10) give information about the loss of quality.
In the literature there are few articles about white chocolate despite its importance and presence
in the market. Some ingredients of the white chocolate formulations can accelerate the loss of
color and others can decrease it. One ingredient, recently made available to the industries, by
Corn Products Company, called GlobeTM Chocosystem, is offered as an inhibitor of the
browning in white chocolate.
The main objective of this research was to determine the kinetics data of transformations that
entail loss of quality in white chocolate, being color the attribute chosen, to be studied over
time or the shelf-life. Three basic formulations were studied, including whey and Globe Choco
System® as ingredients.

MATERIAL & METHODS

For the kinetic studies three formulations were prepared: WCh1 containing 9% of
demineralized whey in the formulation, WCh2 containing 9% of skimmed powered milk and
WCh3 containing 9% Globo Choco System®. Each formulation was completed with 47.0%
sugar, 27.4% cocoa butter, 16% of whole powered milk, 0.3% soya lecithin, 0.2% of PGPR
(polyglycerol polyricinoleate) and 0.1% of aroma.
White chocolate samples were produced in the pilot-plant following the conventional steps [2].
Color: According to the Instruction of colorimeter, model Chroma Meter CR-400/410, Konica
Minolta obtaining the L*, a* and b * values of the Cielab System, at 25 °C.
Calculation of kinetic parameters: For the kinetics study the samples (Table 1) were stored at
10, 20 and 30 °C and 82% RH to accelerate the reaction. For kinetic data it was monitored the
variation of color (values L*, a*, b*). For kinetic parameters calculation first was determined
the order of reaction, by charts of the constant k reaction. The values of ln k were plotted as a
function of inverse temperature (Arrhenius chart). Through the values of the slope of the
straight line, the activation energy (Ea) was determined. The calculation of Q10 was done
according to the Ea as shown in Q10 = 10 ((Ea/(0.46 x T ^ 2)).

RESULTS & DISCUSSION

Initially, the water activity (at 25 °C) of samples was around 0.40 and at the end it was around
0.70. It suggests there was water uptake, change of physical state and loss of quality.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1799
The results were obtained over time with the three formulations of white chocolate stored at 10,
20 and 30 °C and 82% RH.
Kinetic Parameters: “a*” values were selected for kinetic study. The order of reaction was
determined as 2nd order. Values of R2 and equations, activation energy and Q10 for the
formulations (WCh1, WCh2 and WCh3) were presented at Table 1.

Table 1. Values of R2 and equations, activation Energy (Ea) and Q10 for the formulations (WCh1, WCh2,
WCh3), for lost of color of white chocolate
Ea
T (°C) R2 Equation (y) (kcal/ gmol.K) Q10 *
WCh1 25.43 4.0
10 0.856 -0.0001x-0.3618
20 0.989 -0.0003x-0.3655
30 0.918 -0.002x-0.2619
WCh2 38.27 9.3
10 0.863 -0.00008x-0.3439
20 0.965 -0.0002x-0.3447
30 0.958 -0.0007x-0.2964
WCh3 9.25 2.0
10 0.866 -0.0001x-0.4116
20 0.952 -0.0001x-0.4199
30 0.837 -0.0003x-0.3641
*Q10=10((Ea/(0.46 x T^2))

If the activation energy and Q10 increase, the monitored transformation of the food will
accelerate [3]. The results of Table 1 showed the formulation WCh3 (prepared with GlobeTM
Chocosystem, without skimmed milk or whey) had more stability related to color, followed by
formulation WCh1 and WCh2.

CONCLUSION
It can be concluded that white chocolate containing GlobeTM Chocosystem can maintain the
white color longer compared to formulations containing whey and even more in those
containing skimmed milk.

ACKNOWLEDGMENTS: We do hereby thank CNPq [Conselho Nacional de Desen-

volvimento Científico e Tecnológico] and CornProducts Brasil - Ingredientes Industriais.

REFERENCES
[1] VERCET. A. Browning of white chocolate during storage. Food Chemistry. v. 81. p. 371-377. 2004.
[2] BECKETT. S.T. Industrial Chocolate Manufacture and Use. 2.ed. London: Chapman and Hall. 1994.
408p. [2] VERCET. A. Browning of white chocolate during storage. Food Chemistry. v. 81. p. 371-377.
2004. [3] TAOUKIS. P.. LABUZA. T. P.; SAGUY. Kinetics of Food Deterioration and Shelf-Life
prediction. In: The Handbook of Food Engineering Practice. K.J.Valentas. E.Rotstein. R.P.Singh (ed).
CRC Press. p.361-403. 1997.

1800
 Available lysine in powdered infant formula as described by reaction kinetics

I. Schmitza, A. Gianfrancescob, U. Kulozika, P. Foersta

a
Chair for Food Process Engineering and Dairy Technology, Technische Universität München,
Freising/Weihenstephan, Germany (iris.schmitz@wzw.tum.de)
b
Nestlé Research Center, Nestec Ltd., Lausanne, Switzerland,
(alessandro.gianfrancesco@rdls.nestle.com)

INTRODUCTION
Powdered infant formulas are usually produced by spray drying which is a gentle drying
procedure. However, degradation reactions as the loss of available lysine due to the Maillard
reaction with lactose can occur during spray drying of infant formula.[1, 2] Milk proteins contain
relatively high amounts of lysine which is an essential amino acid that can only be metabolized
in the human body if it has a free İ-amino group. However, so far the loss of available lysine
observed after spray drying has not systematically been linked to the processing conditions. As
the product is dehydrated and heated simultaneously during spray drying, the temperature and
concentration of the product change continuously during the drying process. Thus, to describe
the lysine losses during spray drying, the reaction kinetics have to be established for the high
concentration regime at temperature-time conditions relevant for spray drying which has not
been done yet.
The objective of this study is to describe the loss of available lysine in an infant formula model
system at conditions applicable to spray drying, i.e. in a high concentration regime. These
reaction kinetics are established taking into account the concept of water activity, glass
transition and molecular mobility with the aim of obtaining a deeper understanding of the
mechanism of lysine loss in concentrated milk systems.

MATERIALS & METHODS

The model system was prepared by reconstituting skim milk powder, whey protein isolate,
lactose, potassium citrate and sodium hydrogenphosphate in deionized water (all provided by
Nestec Ltd., Switzerland). The solution was freeze-dried, ground after freeze-drying and the
water activity of the obtained powder was adjusted to 0.11, 0.23, 0.33 and 0.43. For the
heating experiments, the samples were transferred to heating cans and heated in a water bath
at 60 – 90 °C for 0.5 – 30 min. Available lysine was determined using the fluorimetric o-
phthaldialdehyde (OPA) method. To measure the glass transition temperature of the
samples, differential scanning calorimetry was used in the modulated mode. Midpoint
temperatures are given. The delay of lactose crystallization was quantified by isothermal
differential scanning calorimetry (DSC). The molecular mobility was evaluated by measuring
the transversal relaxation time T2 with a low resolution 1H-NMR spectrometer

RESULTS & DISCUSSION

Loss of available lysine was observed at all conditions applied. The lysine loss increases with
heating time for all temperatures and water activities (data not shown). A maximum in lysine
loss was determined for all temperatures (Figure 1), while the location of the maximum

11th International Congress on Engineering and Food - Athens, Greece, 2011 1801
depends on the temperature. Rather small lysine losses were measured at 60 °C and the impact
of the water activity was not significant. Raising the temperature to 70 °C leads to significantly
higher lysine losses and a maximum was detected at a water activity of 0.43. The maximum in
lysine loss is shifted to lower water activities at higher temperatures and can be found at
aw 0.23 at 80 and 90 °C.

Figure 1. Loss of available lysine after 30 min of heating as a function of water activity, temperature and
distance from the glass transition temperature.

Taking the physical state of the system into consideration, it becomes obvious that lysine
losses are small in the glassy state. Furthermore, it can clearly be noticed that the maximum in
lysine loss is found in the transition zone from the rubbery to the crystalline state. Under these
conditions, the samples stay for the longest period in the rubbery state. Samples that are
crystalline after 30 min of heating passed the rubbery state faster. This means that the rubbery
state is more reactive regarding lysine loss than the glassy and the crystalline state.

CONCLUSION
Our results show that lysine losses increase with increasing temperature and are dependent on
the water activity. The importance of the physical state of lactose and of the molecular mobility
(data not shown) arises. The kinetics obtained during this study are used to build a model
describing lysine blockage during spray drying. This should allow optimisation of the drying
process, thus limiting the loss of available lysine.

REFERENCES
[1] Meade S.J., Reid E.A. & Gerrard, J.A. 2005. The impact of processing on the nutritional quality of
food proteins. Journal of AOAC International, 88(3), 904-922.
[2] Finot P.A. 1983. Chemical modifications of the milk-proteins during processing and storage -
nutritional, metabolic and physiological consequences. Kieler Milchwirtschaftliche
Forschungsberichte, 35(3), 357-369.

1802
 Kinetic modelling of colour changes during beef roasting

S.M. Goñia,b, V.O. Salvadoria,b

a
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA, CONICET-La Plata),
Fac. de Cs. Exactas - UNLP, 47 y 116, B1900AJJ, La Plata, Argentina.
b
MODIAL, Área Deptal. Ing. Qca., Fac. de Ingeniería, UNLP, 1 y 47, B1900TAG, La Plata, Argentina.
(smgoni@cidca.org.ar, vosalvad@ing.unlp.edu.ar)

INTRODUCTION
During the roasting of beef samples, the muscle suffers several changes associated with its
internal temperature evolution. At the end of cooking beef’s colour is the main indicator of the
doneness as it is perceived by consumers. This way of perceiving doneness by consumers can
lead to safety problems, since there exists evidence that visual appearance does not imply that a
safe temperature, from a microbiology point of view, have been reached ([1]). On the other
hand, mathematical modelling and simulation has been proved to be a useful tool in several
meat processing operations, like hamburger cooking, where calculation of cooking time to
achieve safe products and optimization of the process has been done successfully. Then, the
objective of this work was to develop a simple kinetic model to describe the colour changes
produced during beef roasting. Furthermore we test coupling it to a previous developed and
validated model of beef roasting, which describe simultaneous heat and mass transfer during
the process.

MATERIALS & METHODS

Thin slices (4 mm thickness, 4u4 cm2 lengthuwidth) of beef semitendinosus muscle were used
to obtain colour change information. The samples were packed and then subjected to different
time-temperature treatments (2.5-30 minutes, 40-100ºC) using a thermostatic water bath. After
each heat treatment, instrumental measures of surface colour in the CIEL*a*b* colour space
was obtained, using a MINOLTA colorimeter.

RESULTS & DISCUSSION

In all the treatments the a*-value was lower than the raw ones, the variation was more
pronounced at high temperatures. The L*-value of cooked samples was higher than the value of
raw ones, and the variation was less pronounced at high temperatures and long process time.
Variations in b*-value did not exhibit a clear trend, being in a narrow region.
From this analysis, variations of the a*-value appears to be the most important, since it
determines the change from a pink-red colour of raw beef to a grey-brown colour of cooked
beef; such changes are actually the ones perceived by consumer and can determine product
acceptability and doneness. To describe the variation of a*-value a first order fractional kinetic
model was used (Eq. (1)), being reaction rates (k) correlated with temperature according to an
Arrhenius relationship. An equilibrium value a*f depending on temperature also was obtained
from experiments (Eq. (2)). The model fits well to the experimental data, with an absolute
relative error of 6.45%; parameters were found as k0=1.4482u1010 s-1, and the activation energy
was found as Ea=80.7397 kJ mol-1.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1803
da* (1)
k  a*  a*f 
dt
15.08 (2)
af* a0* 
1  1.84exp  0.17  47.17  T  

Later, the kinetic model was coupled to a previously developed and validated beef roasting
model. Such model considers simultaneous heat and mass transfer during the process, and their
predictions were in good agreement with experimental test ([2], [3]). Since for large samples
can exist large temperature differences between inner-outer regions, also can exist colour
differences in such regions. Then the model allow to obtain a non uniform internal distribution
of a*-values, which is actually observed in large cooked samples, still more when end
temperature is low. For instance, Figure 1 shows the application of the methodology using a
3D irregular domain of one semitendinosus muscle sample. The cooking model was simulated
until the coldest point in the geometry reach a temperature of 72ºC.

Figure 1. Example of the application of the methodology. (a) 3D irregular geometric model; (b) middle
cross-section colour (a*) prediction coupled to whole domain, at different process times.

CONCLUSION
From a model-based framework, the usefulness of beef cooking models describing the relevant
heat and mass transfer mechanisms can be improved incorporating other important issues,
related to safety restrictions, i.e. to reach a given temperature or lethality at certain point, and
also consider quality features, as colour or texture, among others. In this sense, the developed
model, which describes the redness variation during beef roasting, can be used to establish
operating strategies to reach desirables colour values or colour uniformity at end of cooking.
Furthermore, other beef muscles (or muscles of another kind of animal) can be considered
provided that appropriate information of colour variation is known.

REFERENCES
[1] King, N., & Whyte, R. (2006). Does it look cooked? A review of factors that influence cooked meat
color. Journal of Food Science, 71(4), 31-40.
[2] Goñi, S.M. (2010). Simulación y optimización de la cocción de productos cárneos en hornos
convectivos, Ph.D. thesis. Universidad Nacional de Quilmes, Argentina.
[3] Goñi, S.M., & Salvadori, V.O. (2010). Prediction of cooking times and weight losses during meat
roasting. Journal of Food Engineering, 100(1), 1-11.

1804
 Instrumentation of a semi-industrial oven to monitor non-enzymatic browning kinetics

during baking

Courel Mathildea,b, Rega Barbaraa,b, Fehaili Souada,b, Giampaoli Pierrea,b and Bonazzi Catherinea,b

a
INRA, UMR1145 Ingénierie Procédés Aliments, Massy, France (mathilde.courel@agroparistech.fr)
b
AgroParisTech, UMR1145 Ingénierie Procédés Aliments, Massy, France

INTRODUCTION
The objective of this work was to design and characterize an experimental device enabling to
generate reliable kinetic data on thermal reactions occurring during the transformation of
bakery products under real processing conditions. A baking oven was thus developed with the
objective of controlling heat fluxes, which is a determining parameter of baking process.
Sponge cake was used as a model of solid matrix which complex composition and structure is
changing during baking. In this study, 5-hydroxymethylfurfural (HMF) was used as a specific
chemical indicator of both Maillard and caramelization reactions and monitored in the food
matrix as well as in the baking vapors. This work was carried out with the financial support of
the French National Research Agency under the "Programme National de Recherche en
Alimentation et nutrition humaine" project ANR-06-PNRA-023.

MATERIALS & METHODS

Sponge cake preparation: composition and preparation of the dough are fully described in [1].
The baking oven was a static convective batch type oven (Bongard, France, 7.5 kW power, 96
L working volume) generating ambient temperatures of up to 300°C. A frequency variation
system (25 to 50 Hz) provided two convection levels: min and max. K-type thermocouples
were used for cake and air temperature measurements. Cakes’ surface temperature was
measured using an infrared thermometer (Optris CT, Germany). A Hygrox-C2 humidity sensor
(McQueen Cairns International, UK) enabled monitoring air humidity in the oven. Heat flux
sensors gave access to radiative and convective surface heat fluxes, ĭrad and ĭconv (W.m-2),
enabling estimation of hconv (W.m-2.K-1), the convective heat transfer coefficient. It remained
very stable with a mean relative standard deviation of ± 2% during acquisition time. The flux
mapping gave relative standard deviations of reproducibility of 6.5%, 5.5% and 3.0% for ĭconv,
ĭtotal and hconv respectively. Two original sampling devices were specifically designed to carry
out kinetic studies during baking. The sponge cake sampler enabled extracting a pan within 10
s without disturbing the oven thermal environment. The vapor sampling device was based on
the dynamic Head-Space Solid Phase Micro-extraction (HS-SPME) of vapors during baking.
The SPME fibers were analyzed immediately after extraction using GC–MS [1].
Baking conditions: temperature/ventilation were set at 140°C/max (A), 170°C/max (B),
170°C/min (C) and 200°C/max (D) with 30 min total baking time. Cake and baking vapors
were sampled synchronously during 5 min intervals throughout baking. Temperatures (cake
core and surface, air), air humidity, water content, HMF concentration were monitored during
baking. Experiment (B) was done in triplicate for reproducibility estimation of the baking
operation and extent of chemical reaction at all sampling times.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1805
RESULTS & DISCUSSION
Simultaneous changes to HMF concentrations in the cake and baking vapors at a baking
temperature of 170 °C are represented in figure 1-A. After 5 min baking, the concentration of
this compound rose rapidly showing intense production in the cake matrix and volatilization in
the oven. An 8.7% relative error of reproducibility was obtained in the sponge cake which is
very satisfactory and 20.4% in the baking vapors, which corresponds to the reproducibility of
the analytical method.
Three loads of sponge cake (100, 240 and 420 g) were baked independently at 170°C with
maximum ventilation, to check for possible saturation of the SPME extraction fiber. The
chromatographic response of all studied volatile compounds had a linear shape (R2 > 0.9)
versus baking load; the case of acetic acid is shown in figure 1-B. Thus, despite air extraction
for volatile analyzes and partial air renewal in the oven, the conditions prevailing during a
baking operation are steady enough to let the composition of the baking vapors be
representative of the load being baked.
A. Hydroxymethylfurfural B. Acetic acid
60 24 200
HMF in sponge cake
peak area/g DM (E+05)

50 HMF in baking vapors 20 30 min

mmol/g DM (E-04)

160
y = 0,3676 x + 32,721
Peak area (E+06)

2 25 min
40 16 R = 0,9926
120 20 min
30 12 15 min
80 10 min
20 8
y = 0,0382 x + 2,2918 5 min
10 4 40 2
R = 0,9933

0 0 0
0 5 10 15 20 25 30 35 0 100 200 300 400 500
Baking time (min) Sponge cake load (g)

Figure 1. 1-A. Monitoring of HMF concentration during baking at 170°C in sponge cake and baking
vapors. 1-B. Chromatographic response of volatile acetic acid sampled during the baking of different
sponge cake loads at 170°C. DM: dry matter.

CONCLUSION
A semi-industrial convective oven was instrumented and equipped with two original sampling
devices to study the development of thermal reactions like Maillard and caramelization during
the baking of sponge cake under fully controlled thermal conditions. Uniformity and
reproducibility of the heat fluxes in the oven cavity lead to highly reproducible development of
non-enzymatic browning reactions: 8.7% relative variation was obtained for HMF production
in the cakes at six different baking times of a 30 min baking operation. This oven should be of
precious help for further kinetic modeling of complex thermal reactions occurring in solid food
matrix baked under real process conditions.

REFERENCES
[1] Fehaili S. 2010. Développement d’un simulateur de cuisson pour l’étude du couplage entre les
transferts d’énergie et de matière et les cinétiques de réactions de Maillard ayant lieu au cours de la
cuisson de produits céréaliers de type génoise. Unpublished PhD Thesis. Institute of Life and
Environmental Science and Technology, Department of Food Science and Process, Massy, France.

1806
 Degradation of 5-Hydroxymethylfurfural in Malt during Fermentation of Beer

Gül AkÕllÕo÷lu, Burçe Ataç Mogol, Vural Gökmen

Department of Food Engineering, Hacettepe University, 06800 Beytepe, Ankara, Turkey

(gulakillioglu@hacettepe.edu.tr, burcea@hacettepe.edu.tr, vgokmen@hacettepe.edu.tr )

INTRODUCTION
During brewing, Maillard reaction and caramelization occur as roasting proceeds at high
temperatures up to 250 ºC leading to the formation of HMF and furfural [1]. There are several
reports claiming that furfural and HMF inhibit the growth of yeasts, hence cause a decrease in
the yield of ethanol production [2, 3]. On the other hand, HMF was found to be taken up and
converted by the yeast during both aerobic and anaerobic conditions, and the main conversion
product was found to be 5-hydroxymethylfurfuryl alcohol (HMF alcohol) [4]. Preliminary
experiments indicated that HMF levels in beers are relatively low (<5 mg/L) even though
significantly higher levels (>1000 mg/kg) may form in dark roasted malts. Thus the present
study aimed to investigate the mechanism and kinetics of HMF degradation during yeast
fermentation of malt.

MATERIALS & METHODS

Wort obtained from roasted and pale malt was fermented with baker’s yeast (Saccharomyces
cerevisiae) at 30ºC for 24 hours. Sampling was performed at certain time intervals during
fermentation that was carried out both in the presence and absence of sugar. HMF and sugar
concentrations were determined using Shimadzu UFLC system (Shimadzu Corporation, Kyoto,
Japan) and Agilent 1100 HPLC system (Waldbronn, Germany), respectively.

RESULTS & DISCUSSION

HMF content of dark malt was found to be 520 mg/kg. Wort and sweet wort obtained from the
dark malt had HMF contents of 71.99 ± 0.89 and 65.50 ± 1.10 mg/L, respectively. It was
observed that HMF contents of both worts were decreased during fermentation process
indicating that it is utilized by yeast.
HMF data were evaluated with MATLAB® 7.0.1. HMF content of worts showed an
exponential decay as fermentation progressed (Figure 1). Data were fitted to equation
[HMF]=[HMF]0×(exp(-k×t)), where “t” and “k” correspond to time in minutes and rate
constant min-1, respectively. The degradation rate constant of HMF was 0.693×10-2 min-1 and
1.397×10-2 min-1 for wort and sweet wort samples, respectively, indicating that sugar enhances
the activity of yeast. Sucrose was converted into glucose and fructose just in the first hour of
fermentation. Glucose and fructose contents of dark roasted malt decreased more rapidly than
those of pale malt (p<0.05). There was no decrease in the number of viable cells during
fermentation and yeast count at the 24th hour of the fermentation was higher in the presence of
HMF.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1807
 Figure 1. Change of HMF concentration in wort during fermentation.

CONCLUSION
The results of present study do not support that HMF may inhibit yeast growth as claimed by
others. It was observed that HMF was utilized as a carbon source during fermentation besides
glucose and fructose, and its degradation was more rapid than those of glucose and fructose. It
is thought that roasting modifies the malt composition that makes the nutrients more available
for yeasts enhancing their growth.

REFERENCES
[1] Woffenden H.M., Ames J.M. & Chandra S. 2001. Relationships between antioxidant activity, color,
and flavor compounds of crystal malt extracts. Journal of Agricultural and Food Chemistry, 49,
5524-5530.
[2] Palmqvist E., Almeida J.S. & Hahn-Hagerdal B. 1999. Influence of furfural on anaerobic glycolytic
kinetics of Saccharomyces cerevisiae in batch culture. Biotechnology and Bioengineering, 62, 447-
454.
[3] Palmqvist E., Grage H., Meinander N.Q. & Hahn-Hagerdal B. 1999. Main and interaction effects of
acetic acid, furfural, and p-hydroxybenzoic acid on growth and ethanol productivity of yeasts.
Biotechnology and Bioengineering, 63, 46-55.
[4] Taherzadeh M.J., Gustafsson L., Niklasson C. & Liden G. 2000. Physiological effects of 5-
hydroxymethylfurfural on Saccharomyces cerevisiae. Applied Microbiology and Biotechnology, 53,
701-708.

1808
 Thermal inactivation kinetics of L-carnitine

Polyvios Prokopioua, Athanasia M. Goulab, Nikolaos G. Stoforosc

a
Pipis Farm Ltd., Nicosia, Cyprus (polyviosprokopiou@hotmail.com)
b
Department of Food Science and Technology, Faculty of Agriculture, Aristotle University of
Thessaloniki, Greece (athgou@agro.auth.gr)
c
Department of Food Science and Technology, Agricultural University of Athens, Greece
(stoforos@aua.gr)

INTRODUCTION
L-carnitine is a quaternary ammonium compound biosynthesized in living cells from the amino
acids lysine and methionine. Its primary function is to facilitate the transport of activated long
chain fatty acids from the cytosol into the mitochondria and, thus, it is essential for the
production of energy from lipids. In addition, L-carnitine helps to remove toxic compounds
from within the cells. Although small amounts of L-carnitine can be synthesized by adults, the
majority of L-carnitine needed in humans is taken through food consumption. Exogenous
supply of carnitine is mainly supplied by foods of animal origin, where its concentration varies
between 8 and 530 mg/kg of dry mass, whereas fruit and vegetables contain very little, if any,
L-carnitine [1]. The development of accurate models able to predict the behavior of substances
under specific environmental conditions is of main importance for the food industry and relies
on the estimation of appropriate kinetic data. However, studies on thermal inactivation of L-
carnitine have not been found in the literature. Thus, the objective of this work was the kinetic
study of thermal inactivation of L-carnitine. Existing procedures for kinetic parameter
estimation are normally based on experiments under isothermal conditions. However,
temperature may vary extensively throughout a thermal process. A few works approach the
estimation of inactivation kinetics considering time-varying temperature conditions [2]. So,
another task undertaken in this study was to develop a procedure for thermal inactivation
kinetics determination, from dynamic temperature profile experiments.

MATERIALS & METHODS

The inactivation experiments of L-carnitine took place in glass Pasteur pipettes immersed in a
water/oil bath set at specified temperatures (80, 85, 90, 95, 100, 110, 120, and 130°C) selected
after preliminary experiments. The L-carnitine content was spectrophotometrically determined
[3]. DT values at each temperature tested were calculated with the progressive use of the
experimental remaining concentration data (Eq. (1)) and afterwards these DT values were used
for the calculation of the z value (Eq. (2)) by successive linear regressions.
t (1)
log C log C o 
DT
Tref  T (2)
log DT log DTref 
z
where Co is the initial L-carnitine concentration, C is the L-carnitine concentration at time t, DT
is the decimal reduction time at temperature T, z is the thermal resistance constant, and Tref is a
reference temperature. Furthermore, remaining L-carnitine concentration data were collected
during a non-isothermal experiment for a step-wise increasing temperature profile.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1809
RESULTS & DISCUSSION
Data on remaining L-carnitine concentration against processing time as a function of
processing temperature are shown on Fig. 1. The parameters D120°C and z were calculated equal
to 50.6 min and 30.2°C, respectively.

-2
log C (C in g/100 mL).

-3

80°C 85°C 90°C 95°C

-4
100°C 110°C 120°C 130°C

-5
0 100 200 300 400 500 600
t (min)

Figure 1. Experimental (symbols) and predicted (lines) values of remaining L-carnitine concentration as
a function of processing time and temperature.

Similar values were obtained from the measurements during the dynamic temperature profile
experiment. For this case, D120°C and z values were found equal to 48.8 min and 29.9°C,
respectively. The two methods resulted in not significantly different results. In addition, both
methods yielded precise and accurate predictions. Correlation between experimentally
determined concentrations of L-carnitine after non-isothermal treatment and those calculated
by means of kinetic parameter estimates form dynamic and isothermal data gave R2 and SSE
values of 0.999 and 8.19, respectively, for the dynamic method and 0.998 and 10.28 for the
isothermal experiments.

CONCLUSION
Based on the agreement between the parameters estimated using isothermal or non-isothermal
temperature profiles, and given the reduced number of experimental data required by the latter
approach, kinetic parameter estimation from experiments at dynamic conditions is
recommended. Although in the present study L-carnitine thermal inactivation followed first
order kinetics and the methodology used was focused on such kinetic behavior, deviations from
first order kinetics can be handled.

REFERENCES
[1] Seline K.-G. & Johein H. 2007. The Determination of L-Carnitine in Several Food Samples. Food
Chemistry, 105(2), 793–804.
[2] Van Boekel M. 1996. Statistical Aspects of Kinetic Modelling for Food Science Problems. Journal of
Food Science, 61, 477–485.
[3] Schafer J. & Reichmann H. 1989. A Spectrophotometric Method for the Determination of Free and
Esterified Carnitine. Clinica Chimica Acta, 182, 87–94.

1810
 Quality degradation of butterhead lettuce: the performance of General Stability Index
(GSI) modified methodology

María Victoria Agüeroa, Sara Inés Rourab

a
Grupo de Investigación en Ingeniería en Alimentos (GIIA). Facultad de Ingeniería. UNMdP. CONICET,
Mar del Plata, Argentina. (mvaguero@fi.mdp.edu.ar)
b
GIIA. Facultad de Ingeniería. UNMdP. CONICET, Mar del Plata, Argentina. (sroura@fi.mdp.edu.ar)

INTRODUCTION
Lettuce shelf life is a dynamic period in which microbiological, chemical, enzymatic and
physicochemical reactions simultaneously take place affecting several quality aspects. Each
quality index presents particular behavior during lettuce storage. Moreover, changing storage
conditions affects differentially each index. These facts make the evaluation of shelf-life an
arduous task. GSI (General Stability Index) method [1] and its modified version [2] are
valuable tools that allow the simultaneous evaluation of most relevant quality indices
expressing such variations as a single value. The objective of the present work was to
characterize the kinetic evolution (specially reaction order and constant rate) of GSI for
butterhead lettuces stored under two different conditions commonly found during its
distribution from farm to vegetable processors: optimal (0-2ºC, 97-99% relative humidity) and
suboptimal (0-2ºC, 70-72% relative humidity). Each lettuce section (external, middle and
internal) was analyzed independently.

MATERIALS & METHODS

Quality indices were: relative water content (RWC), water content (WC), free water (FW),
bound water (BW) and the ratio free to total water (FW/TW) as physiological indices; total
chlorophyll content (C) as greenness index; ascorbic acid content (AA) as nutritional index,
total microbial counts (MC) as microbial index and overall visual quality (OVQ) as sensorial
index. Sampling days were: 0, 2, 5, 8, 13, 16 and 20; and 0, 1, 2, 3, 4 and 5 for lettuces stored
under optimal or suboptimal conditions, respectively. GSI was calculated for each lettuce
section and each storage condition following the methodology suggested by Ansorena et al.
(2009). GSI was modeled according to: d(GSI)/dt=-k(GSI).
Finally, it was established the dependence of kinetic parameters (n and k) with section and
storage condition. Statistical analyses were carried out using SAS software version 9.0 (SAS
Institute 2002). Adjustment of equations was done using SYSTAT 5.0 (SYSTAT Inc, 1992).

RESULTS & DISCUSSION

The lettuce section and the storage condition significantly affected the composition of the
tetrad that best represent the quality evolution during storage. The tetrads selected for lettuces
stored under optimal conditions were: (RWC, FW, C, AA), (RWC, WC, C, AA) and (RWC,
FW, WC, AA) for external, middle and internal sections, respectively; and for lettuces stored

11th International Congress on Engineering and Food - Athens, Greece, 2011 1811
under sub-optimal conditions, were: (RWC, WC, AA, MC), (RWC, WC, TC, AA) and (RWC,
BW, WC, MC) for external, middle and internal sections, respectively.
Regardless of the lettuce section or the storage condition, the evolution of GSI was
characterized by a first order kinetic, with a rate constant (k) greatly affected by both factors.

1 kI = 0.05 day-1
0,8

0,6
kI = 0.04 day-1
GSI

0,4
kM = 0.25 day-1 kM = 0.05 day-1
0,2
kE = 0.44 day-1 kE = 0.11 day-1
0
0 5 10 15 20

Time (days)
Figure 1. GSI evolution in external (Ɣ and ż), middle (Ŷ and Ƒ) and internal (S and U) lettuce sections
during storage of lettuce heads under different relative humidity conditions: 97 – 99 %HR (- - - -) and 70
– 72 %HR ( ).

These results supported the great impact of relative humidity condition on butterhead quality
and shelf life. The higher impact of humidity condition occurred in external and middle
sections with a 4.2-4.9 fold increase in the k value, while the internal section only increases 1.3
fold in sub-optimal condition respect the optimal one. Differences observed in k values from
different lettuce sections could be attributed to both the degree of tissue development and the
differential exposure to environment. In this way, internal leaves are younger than external
ones and this could imply different physiological behavior. Moreover, due to lettuce plant
morphology, the internal leaves are wrapped by middle and external leaves, protecting them
from environment stress.

CONCLUSION
The modified GSI methodology was successfully applied to evaluate the quality degradation of
butterhead lettuce under different postharvest conditions. The GSI was composed by different
indices depending on both lettuce section and storage condition. However, as GSI is a
dimensionless parameter, comparisons of GSI evolution could be done. First order kinetic was
found for all situations but the rate constant was greatly affected by both factors. The
methodology simplifies the quality evolution quantification because it recommends the
determination of only four indices, and simplifies the comparisons between different storage
conditions.

REFERENCES
[1] Achour, M. (2006). A new method to assess the quality degradation of food products during storage.
Journal of Food Engineering, 75 (4), 560-564.
[2] Ansorena, M.R., Goñi, M.G., Agüero, M.V., Roura, S.I., & Di Scala, K.C. (2009). Application of the
General Stability Index to assess the Quality of Butter Lettuce during Postharvest storage by a multi-
quality indices analysis. Journal of Food Engineering, 92, 317-323.

1812
 A MALST method comparison over univariate kinetic modelling for determination of

Shelf life in cereal snack of dried apples

J. Saavedraa,b, A. Córdovaa, C.Quezadaa.

a
Research Group on Chemometrics, Department of Food Engineering, Pontificia Universidad Católica de
Valparaíso. (jorge.saavedra@ucv.cl)
b
Centro Regional de Estudios en Alimentos Saludables (CREAS). Valparaíso, Chile.

INTRODUCTION
Formal determination of Shelf Life is a key factor in the research and development of food,
since provides information regarding the time that the product aptly retains its attributes. This
determination is usually performed by measurement of quality attributes [1], alternating
accelerated ageing methods under extreme conditions, being methodoloy proposed by [2], the
mostly used. However, the determination becomes complex when multiple attributes must be
studied simultaneously since each one has an own specification related with the date of
expiration, being useful in these cases the use of multivariate statistical techniques. In this
sense, Accelerated Shelf Life Testing methods have incorporated multivariate tools, called
Multivariate Accelerated Shelf Life Testing (MALST). In this context, the objective of this
research was comparatively assessing Shelf Life of an absolutely new product in market snack-
cereal of dried apple type, through traditional univariate method and multivariate accelerated
method.

MATERIALS & METHODS

Samples of a new product in the market of dried Apple snack type from an exporting
agribusiness (Maule, Chile) packed in multi-laminated bags were incubated at 18 ° C, 25 ° C
and 35 ° C, for 18 months. Quality attributes considered: Aw, humidity, content of SO2,
sensory attributes, and color CIE-Lab. Data were modeled by univariate degradation kinetics [2,
3] and multivariate analysis was performed with PCA analysis, in order to model multivariate
kinetics, according methodology proposed by [5]. All calculations and adjustments were made
with SIMCA-P+ 12, Sigmaplot 11 and Excel 2003.

RESULTS & DISCUSSION

Graphic Scores (Figure 1) shows the evolution of the attributes used at three temperatures.
Model retained 2 Principal Component (PC) explaining 83.1% of the total variability (PC 1:
68% and PC2: 16.2%). PCA ordered the variability of samples based on the time through the
first component (t1). Inspecting the graphics of contribution it could be appreciated the
variability explained by the second computer (t2) explains different behavior for 3 temperatures
profiles of storage. Thus, for the treatment of 18°C, contribution is mainly explained by
attribute Aw, while at 25°C, the color variables of and SO2 content acquired greater importance.
At 35°C the greatest contributions were associated with moisture and texture, at final incubation
times. The above, would imply that model reflected in terms of variability, biochemical
phenomena associated with the deterioration of the product.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1813
 Figure 1. Scores Plot for 3 storage temperatures

From the specification values of the product (not shown values) and the loadings matrix of the
first PC related with time, was obtained the cut-off criteria (tc), which in this case was 0.2183.
It can be observed first order kinetic degradation for all storages conditions. Thus, shelf-life was
18.2 months for storage to 18 ° C, 18.1 months at 25 ° C and 15.5 months at 35 °C. By
comparing this value with the obtained by univariate kinetics, where the degradation only
contemplates humidity as attribute, shelf-life of the product was 17.4 months, existing a
difference of a few weeks in estimating by one method and other.
Due to little difference of Shelf Life at 18°C and 35°C, it could be inferred the high stability of
the product.

CONCLUSION
MALST methodology could estimate simultaneously deterioration of quality attributes of the
product, showing the interactions that occur between them.

REFERENCES
[1] Van Boekel, M., 2008. Kinetic Modeling of Food Quality: A Critical Review, in Comprehensive
reviews in Food Science and Food Safety 7, Institute of Food Technologists p. 144-158.
[2] Labuza, T., 1982. Shelf Life Dating of Foods. Food and Nutricion Press, Inc. Westport, Connecticut,
U.S.A
[3] Man,C. & Jones, A.,1999. Shelf Life Evaluation of Foods. Aspen Publishers, Inc, Gaithersburg.
[4] Sjostrom, M., Wold, S., Lindberg, W., Persson, J.A., Martens, H., 1983. A multivariate calibration
problem in analytical chemistry solved by partial least-squares models in latent variables, Analytica
Chimica Acta, Volume 150, 1983, Pages 61-70.
[5] Pedro & Ferreira, 2006: Multivariate accelerated shelf-life testing: a novel approach for determining
the shelf-life of foods. Journal of Chemometrics, 20, 76–83.

1814
 Modulation of thermal inactivation of protease during enzymatic hydrolysis of salmon

muscle

Pedro Valencia, Noé Bustos, Sergio Almonacid

Universidad Técnica Federico Santa María, Valparaíso, Chile (pedro.valencia@usm.cl)

INTRODUCTION
Modulation consists in the protection or destabilization of an enzyme by substrates or products
present in reaction [1]. This is a major concern for the hydrolysis of insoluble proteins using
proteases because enzyme inactivation is a significant phenomena in this process and adsoption
of enzyme on substrate suggests that native conformation of enzyme is stiffened causing a
stabilization of activity. The objective of this work was to compare thermal inactivation of
Alcalase under non-reactive and reactive conditions during the process of enzymatic hydrolysis
of the substrate salmon muscle protein.

MATERIALS & METHODS

In non-reactive conditions Alcalase was mixed with 100 mM phosphate buffer pH 8.0. In
reactive conditions Alcalase was mixed with a blended muscle slurry prepared from salmon
fillets and the same phosphate buffer following the method of Krinstinsson and Rasco [2]. In
both conditions the mixture was maintained in a temperature-controlled bath at 60ºC and
constant pH 8.0 during the whole experiment. Samples were withdrawn, cooled and assayed
for Alcalase activity with azocasein method.

RESULTS & DISCUSSION

Thermal inactivation of Alcalase was tested under reactive and non-reactive conditions, in the
presence and absence of the salmon muscle as protein substrate. The effect of proteic substrate
on the thermal inactivation of Alcalase is observed in Figure 1. A protective effect is observed
when Alcalase was maitained in the presence of salmon muscle proteins at 1% and 5% being 4-
fold and 7-fold more stable than in non-reactive condition. Half-life of enzyme activity in non-
reactive conditions was 33 min and increased until 142 and 233 min in the presence of 1% and
5% of salmon muscle protein. These experiments reveal the protective effect of the substrate
upon the thermal inactivation of Alcalase. The hypothesis is positivelly contrasted with this
evidence. The protective effect during the thermal inactivation of Alcalase suggests that
enzyme tightly attaches to muscle proteins during hydrolysis decreasing the loss of its three-
dimensional conformation. This implies that the adsorption of the enzyme is a step to be taken
into account when modeling the hydrolysis of insoluble protein by-products. Thermal
inactivation experiments in reactive conditions were done during the hydrolysis of salmon
muscle protein, so substrate concentration decreased during reaction. Degree of hydrolysis
during reactive condition experiments reached about 15% in 200 min. This means that product
generated during hydrolysis of proteins could have a modulating effect on the enzyme. This
effect was not quantified and the possibility that peptides formed from hydrolysed proteins

11th International Congress on Engineering and Food - Athens, Greece, 2011 1815
modulates thermal inactivation of enzyme cannot be supported nor discarded with these results.
New experiments must be designed for this purpose.

Figure 1. Thermal inactivation of Alcalase at 60ºC in non-reactive (triangles) and reactive conditions
with 1% (circles) and 5% (squares) of salmon muscle protein. Lines represent first-order model.

CONCLUSION
Thermal inactivation of Alcalase during hydrolysis of salmon muscle protein was clearly
exposed. The protective effect of substrate on the thermal inactivation of Alcalase during the
hydrolysis of salmon muscle protein was demonstrated. Thermal inactivation of Alcalase was
decreased in the presence of salmon muscle proteins. This observation suggests that enzyme
tightly attaches to this substrate during hydrolysis avoiding the loss of its three-dimensional
conformation. In this way mechanisms and conceptual mathematical models for hydrolysis of
proteins must consider adsorption, thermal inactivation and modulation components for proper
enzyme reactor design.

REFERENCES
[1] Illanes A. 2008. Enzyme Biocatalysis: Principles and Applications. Springer Science + Business
Media B.V.
[2] Kristinsson H., Rasco B. 2000. Kinetics of the hydrolysis of Atlantic salmon (Salmo salar) muscle
proteins by alkaline proteases and a visceral serine protease mixture. Process Biochemistry 36(1-2),
131-139.

1816
 Determination of aflatoxin M1 in raw milk by HPLC marker as evidence of cattle-food

storage conditions from the herd suppliers of a dairy company in the city of Valledupar

Fragoso E.a, David T. a, Romero S.a, Ospino H.a

a
Universidad de Santander, Valledupar, Colombia (eliamercedes@hotmail.com)

INTRODUCTION
The production of mycotoxins in human or animal food represents a potential danger for health
and milk production. Colombia is a country with large agricultural and livestock activities, and
these areas are seriously affected by the presence of aflatoxins [1]. The contamination by
aflatoxin M1 in milk and its derivatives varies between 40% and 60% and is given by the
presence of aflatoxin B1 in food and feed commodities for consumption of such cattle, which
has become a problem with global significance [2]. To contribute to the monitoring of these
mycotoxins in Colombia, a study on thirty herd providers from a dairy company was carried
out in the city of Valledupar to determine the concentration of aflatoxin M1 in raw milk by
HPLC as evident marker of cattle-food storage conditions.

MATERIALS & METHODS

This research is descriptive. The population consisted of 97 herds suppliers of a dairy company
in the city of Valledupar. The sample used consisted of the 30 herd .The sampling was carried
out for convenience. The research was conducted in three phases: In the first stage data were
organized in the forms of acceptance to create access routes herds. Subsequently conducted an
inspection of the storage in herds for which they filled out the health inspection form. In the
second phase proceeded with the test sample of 250g of the food field collecting 5 points
equidistant from storage. The samples were transported to the laboratory of food science at the
University of Santander. Then processed the samples obtained by direct seeding in the PDA
medium. It took 10 g sample of 250g collected in each of the points of food stored in herds to
seek the fungal growth. Once growth was obtained microorganisms were replicated to PDA
agar for isolation and identification. In the third stage were collected on the platform of receipt
of the dairy raw milk 500 ml of the thirty herds under study obtained a sample. Subsequently
refrigerated and were transported to the toxicology laboratory at the National University where
they were processed by HPLC technique for determination of aflatoxin M1. The limit of
quantification LOQ of the technique was from 5ng/L-1 and the limit of detection (LOD) was
less than 5ng/-1 concentration.

RESULTS & DISCUSSION

Storage conditions in the herds are not adequate for this purpose, which was corroborated with
data from the inspection report, according to which 100% showed clearly identified hazards of
contamination of food intended for livestock consumption. Similarly, 68% do not have an
exclusive site for storage of food for cattle, given that most are exposed to environmental
conditions that promote the spread and growth of A. flavus [3]. Of direct seeding in stored food
PDA agar growth was obtained corroborating mycelial yeasts and inadequate food storage

11th International Congress on Engineering and Food - Athens, Greece, 2011 1817
conditions, this procedure was isolated from the fungus Aspergillus flavus aflatoxin B1
producer, but can’t relate strictly the presence of this fungus with aflatoxin M1 concentrations
found in milk samples for this influence as other factors such as the aw [4].

Figure 1.6. Comparison of concentration levels between herds AFM1 (minimum and maximum)

The found concentrations of aflatoxin M1 in raw milk of 30 herds were within the levels
permitted by the Colombian technical standard ICONTEC, but it would be limiting if you want
to export the product since it significantly exceeds the maximum established for that purpose.
The upper limit was obtained 160.2ng / L (figure 1.6) followed by 86.9ng / L. These according
to this study would be the maximum to which consumers would be exposed as the other values
were lower.

CONCLUSION
Both yeast and mycelial growth were obtained from direct seeding; mycelial growth was
consistent with the producer of aflatoxin B1 fungus Aspergillus flavus. In the determination of
AFM1 concentrations in raw milk, samples yielded results that found concentrations of
aflatoxin M1 in the raw milk of 30 herds within the levels allowed by ICONTEC. The
threshold obtained was 160.2 ng / L followed by 86.9 ng/L. These, according to this study,
would be the maximum limits to which consumers would be exposed as the other values were
lower. According to the results of this study it is recommended to implement the determination
of aflatoxin M1 in raw milk produced in the department of Cesar, in order to verify the real
situation regarding this toxin and thus exercise control and surveillance measures which ensure
the welfare of the consumer population.

REFERENCES
[1] Díaz, G. (2005) Aflatoxina M1: un carcinógeno de potencial presencia en la leche. En: Seminario
nacional en producción y sanidad bovina, Secretaría de agricultura y desarrollo rural. Mayo 18–20:
Bogotá, Colombia. Universidad Nacional de Colombia.
[2] Cullen J. M. & Newberne P. M. 1993. Acute hepatoxicity of aflatoxins In: The toxicology of
Aflatoxins, human health, veterinary and agricultural significance. ISBN 0-12-228255-8 Press Inc,
Baltimore, MD. 3-20.
[3] Gimeno, A. 2002 Principales factores condicionantes para el desarrollo de los hongos y la producción
de micotoxinas.
[4] Gimeno, A. & Martins, M. (2001) Residuos de aflatoxina M1 y otras micotoxinas en leche y
derivados; control y recomendaciones. En: XVIII seminario g-temcal organizado por g-temcal/danone
Portugal Septiembre 28 Lisboa, Portugal.

1818
 Use of a Poisson-gamma regression model to assess the process hygiene criterion for

Enterobacteriaceae on Irish sheep carcasses

Ursula Gonzales-Barron and Francis Butler

Biosystems Engineering, UCD School of Agriculture, Food Science and Veterinary Medicine, University
College Dublin, Dublin 4, Ireland (ursula.gonzalesbarron@ucd.ie; f.butler@ucd.ie)

INTRODUCTION
Historically, in the development or evaluation of sampling plans, two simplifying assumptions
have been always made: that the true concentration of microorganisms is log-normally
distributed within the batch, and that the variance of the samples within a batch is constant.
Gonzales-Barron and Butler [1] showed however that neither of these assumptions necessarily
hold, and in fact they demonstrated that the Poisson-gamma distribution is by far more
appropriate than the Poisson-lognormal or lognormal when modelling low microbial counts in
foods. In the present study, we model the within-batch and between-batch variability in
Enterobacteriaceae counts sampled from pre-chill sheep carcasses using a random-effects
Poisson-gamma regression model, and we use this model in assessing the actual performance
of the process hygiene criterion stipulated in EC No 2073/2005.

MATERIALS & METHODS

Plate count data was available for twenty pre-chill sheep carcasses swabbed on each of the four
sampling visits to five Irish abattoirs (n=400, j=20 batches). The between-batch and within-
batch variability in Enterobacteriaceae sampled from pre-chill sheep carcasses were modelled
using a Poisson-gamma regression model with correlated random effects for the log average
[log mj] and the log dispersion measure [log kj] of the within-batch gamma distributions. In this
model, the random effects between the log average and log dispersion uj and vj, were assumed
to be realisations of a bivariate normal distribution with mean [0,0] and covariance matrix Ȉ.

RESULTS & DISCUSSION

The random effects of the two parameters of the within-batch true distributions (gamma) were
moderately correlated (r=-0.62): the higher the contamination level within a batch, the lower
the dispersion (i.e., the lower the variability). Therefore, there is no reason to assume that the
variability in Enterobacteriaceae counts is approximately constant and independent of the
contamination level (i.e., within-batch mean). According to the model, when five samples are
taken from a batch, only 0.93% (95% CI: 0-4.08%) of the batches of pre-chill sheep carcasses
produced in Ireland would be found to be above the m limit of 1.5 log CFU/cm2 while virtually
no batches would be found to be above the M limit of 2.5 log CFU/cm2.
The Poisson-gamma model was also used in the assessment of the operating characteristic
(OC) curve of the acceptance sampling plan set by EC 2073/2005 (Figure 1). Along the
average values of probability of batch acceptance, the OC curve presents the 95% confidence
intervals that stem from the uncertainty in the dispersion factor sampled for a batch of a given
mean. At the consumer’s risk of 5% (5% probability of accepting a batch that exceeds a level
that poses an unacceptable risk to the consumers), batches of 250 CFU/cm2 mean (95% CI:

11th International Congress on Engineering and Food - Athens, Greece, 2011 1819
150-400 CFU/cm2) may still pass the hygiene criterion. Now suppose that this hygiene
criterion has been developed under the two-class sampling plan, with lognormal assumption
and a constant standard deviation of, say, 0.8 CFU/cm2. In that case, the acceptance sampling
plan m=1.5 log CFU/cm2 and n=5, would have been set to have a 5% probability of accepting
batches of average higher than ~140 CFU/cm2. The Poisson-gamma therefore is more
conservative than the lognormal (two-class sampling plan) as for the consumer’s acceptable
level of safety established by the latter (140 CFU/cm2 at 5% consumer’s risk), a higher sample
size (and/or a lower m limit) would be required to ensure it according to the random-effects
Poisson-gamma approach. To attain a consumer’s level of protection of accepting batches of at
least 140 CFU/cm2 on average with a probability of 5%, a sampling plan could be set at n=5
and m=55 CFU/cm2 (Figure 1). Under this sampling plan, 1.66% (95% CI: 0-19.6%) of the
batches of pre-chill sheep carcasses produced in Ireland would prompt revision of the
production process and hygiene practices. Although not shown comparatively, it is evident
from this analysis that the higher the between-batch heterogeneity in the dispersion measure,
the least effective will be the acceptance sampling plan or hygiene criterion.
1.0
0.9 m=1.5 log CFU/cm²,
n=5 (Micro. criteria)
P(samples' mean < m limit)

0.8
0.7 m=55 CFU/cm², n=5
0.6 (Variables sampling
plan)
0.5
0.4

0.3
0.2
0.1
0.0
10 100 1000
2
Within-batch average (CFU/cm )

Figure 1. Operating characteristic curves of the microbiological criteria (m=1.5 log CFU/cm2; n=5) and a
variables sampling plan (m=55 CFU/cm2; n=5) testing Enterobacteriaceae counts on a batch of pre-chill
sheep carcasses. 95% CI originate from accounting variable within-batch dispersion measures at a given
within-batch average.

CONCLUSION
A new Poisson-gamma modelling framework has been applied for the first time in assessing
the performance of the EC 2073/2005 process hygiene criterion for Enterobacteriaceae in pre-
chill sheep carcasses. A clear advantage of the random-effects Poisson-gamma model is that no
assumption in relation to constant within-batch variance has to be made, and therefore
probabilities can be estimated with confidence intervals, which arise from accounting variable
within-batch dispersion measures at a given within-batch average. The Poisson-gamma model
appeared to be more conservative than the lognormal in relation to variable sampling plans.

REFERENCES
[1] Gonzales-Barron U. & Butler F. 2011. Characterisation of within-batch and between-batch variability
in microbial counts in foods using Poisson-gamma and Poisson-lognormal regression models. Food
Control (2011), doi:10.1016/j.foodcont.2011.01.028

1820
 Improvement of harvesting and processing of cultivated fresh water prawn

(Macrobrachium rosenbergii)

T. C. A. Silva, L. S. Arrieche

Federal University of Espirito Santo , São Mateus, Brazil (leonardoarrieche@ceunes.ufes.br)

ABSTRACT
The availability of water and climate allow promising prospects for the industrialization of
shrimp produced in confinement in Brazil. The challenge lies in maintaining the postharvest
quality, ensured by the disposal of the product properly, since, after the capture and slaughter,
the fish undergoes changes related to the biochemical, chemical, microbiological and also the
loss of nutrients, until the complete deterioration.
This work will analyze the process of improvement of freshwater species Macrobrachium
rosenbergii, to introduce the HACCP (Hazard Analysis and Critical Control Points). The need
for the establishment of the HACCP is related to food security, increased competitiveness of
product marketing and product with uniform standards of quality. Production will be evaluated
at the Cooperativa dos Aquicultores do Espírito Santo – CEAQ, Brazil, from the harvest of
shrimp to delivery of the product in the Cooperative.
The steps cover the description of the processes of harvest, washing, immobilization and
storage relating them to fit with the good manufacturing practices ideals. Critical limits are
established based in the process measures as well as microbiological and physical-chemical so
that they identify control points and critical control points. Routine monitoring, verification
systems and the establishment of corrective actions may be created from these critical limits
and critical control points identified and propose a schedule for improvements in the
beneficiation. In a second step, we intend to search for new chemical additives for improving
the freezing process that can add the greatest value to the product, resulting in the increased
shelf life of shrimp.
We expected to contribute to the generation of scientific and technological knowledge in order
to effectively meet the practical needs of efforts aimed at the industrialization produced in the
state of Espírito Santo, as the freshwater shrimp farming is presented as an alternative
profitable and high growth potential due to its commercial acceptance.

CONCLUSION
The adoption of the HACCP system in shrimp farms has the advantage of providing a high
level of security to the shrimp farm, allow work proactively and not reactively, also contributes
to increased productivity and reduced costs by reducing rework and waste, permits a more
effective monitoring of product quality at lower cost, by the supervisory authorities and
customers.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1821
1822
 Assessing the conditions of milk production on farms based on family farming

Maria da Penha Piccolo Ramosa, Francisca C.N.N.Silvab, Luciana Oliveira de Fariñac, Cláudia Lúcia de
Oliveira Pintod

a
Universidade Federal do Espírito Santo, UFES, São Mateus, Brazil (penhapiccolo@ceunes.ufes.br)
b
Agente de Desenvolvimento Rural, Incaper, São Mateus, Brazil
c
Universidade Estadual do Oeste do Paraná, UNIOESTE, Cascavel, Brazil
d
Empresa de Pesquisa Agropecuária de Minas Gerais, Epamig, Viçosa, Brazil

INTRODUCTION
In Brazil, the milking activity is one of the main sectors of income generation and tax
collection. Given this economic importance and problems related to the quality of milk in
Brazil, the Ministry of Agriculture, Livestock and Supply has approved Instruction 51, which
establishes physicochemical and microbiological tests as well as the Ministry of Agriculture
requires the product cooling immediately after milking on the farm and its delivery in bulk to
dairies [1]. According to data from Embrapa Gado de Leite [2], most milk producers in Brazil
can be classified as small or medium, with daily production from 50 to 100 L, and they are
typically based on family farming. In particular, the state of the Espirito Santo contributes
approximately with 2% of the national milk production, and in São Mateus, a medium town
located in the north of the State, milk production has its origin mainly in rural areas, based on
family farming. These places possess traditional milkmaid cattle, with animal races of low
potential (average of 3.9 L/milk/day/cow) and few animals in lactation per property. Good
Farming Practice (GFP) includes procedures that intervene directly with the production, quality
and harmless of raw milk. It Includes procedures for health control of the mammary gland,
health of the flock, hygiene in milking and environment of production, quality of the water and
hygiene and health of the milking workers. In this sense, the main objective of this study was
to evaluate the hygienic conditions of milk production on farms of São Mateus/ ES/ Brazil, in
order to propose improvements for the sector.

MATERIALS & METHODS

The research was accomplished in 30 rural properties and data was collected by a questionnaire
previously elaborated on the basis of Good Practice requirements, concerning the Farming
Legislation of the Brazilian Agriculture Ministry. Questions included were type of milking,
hygienic habits, health conditions of the milking workers, control of mastitis, resources of
water and water treatment as well. In all properties, manual milking procedure was employed.
The data were analyzed using response values in relative frequency (%).

RESULTS & DISCUSSION

Responses obtained through the questionnaires to farmers from São Mateus town, in the State
of Espírito Santo, they showed a picture very similar to that one observed in other regions of
Brazil [3, 4, 5]. In this study, results showed that only 40% of the milking man used to wash

11th International Congress on Engineering and Food - Athens, Greece, 2011 1823
theirs hands and arms before milking. Thirteen percent were accustomed to use antiseptic
solutions after washing hands and only 4% of the workers used to pass through periodic health
examinations. In none of the properties pre-dipping and post-dipping procedures were carried
out. Equipment cleaning was performed immediately after milking, however, without usage of
products for sanitization. Mastitis control was not performed in 94% of the properties and none
of them accomplished the California Mastitis Test (CMT), which is an important tool in the
detection of subclinical mastitis. The water used for hygienic cleaning of the equipment,
utensils and installations was originated from springs (33%), wells (50%) and local stations of
water treatment (17%). Irregularities detected in these properties can seriously compromise the
quality and safety of raw milk and dairy products.

CONCLUSION
This study was useful to warn the producers about the irregularities and to propose changes for
qualification in GFP. Gradual implementation of improvements will be introduced in order to
adjust the procedures to current legislation. This will reduce profit losses by industry
devolutions, promote increase of the familiar income and contribute to guarantee the
alimentary security of the population as well.

REFERENCES
[1] Brasil. Ministério da Agricultura, Pecuária e Abastecimento. Departamento de Inspeção de Produtos
de Origem Animal. Instrução Normativa nº 51, de 18 de setembro de 2002. Coleta de leite cru
refrigerado e seu transporte a granel. Diário Oficial da República Federativa do Brasil, n. 172, p. 8-13,
20set. 2002a. Seção I.
[2] Embrapa Gado de Leite. Estatísticas do leite. Juiz de Fora, MG, 2007.
[3] Guerreiro, P.K.; Machado, M.R.F.; Braga, G.C.; Gasparino, E.; Franzener, A.S.M. Qualidade
microbiológica de leite em função de técnicas profiláticas no manejo de produção. Ciência e
Agrotecnologia, Lavras, v. 29, n. 1, p. 216-222, jan./fev. 2005.
[4] Nero, L.A.; Viçosa, G.N.; Pereira, F.E.V. Qualidade microbiológica do leite determinada por
características de produção. Ciência e Tecnologia de Alimentos, Campinas, v. 29, n. 2, p. 386-390,
abr.-jun. 2009.
[5] Pedrico, A.; Castro, J. G.D.; Silva, J.E. C.; Machado, L.A.R. Aspectos higiênicos- sanitários na
obtenção do leite no Assentamento Alegre, município de Araguaína, TO. Ciência Animal Brasileira,
v. 10, n. 2, p. 610-617, abr./jun. 2009.

1824
 Regeneration of Frying Oils By Using Adsorbent Resins

Neslihan GÖNCÜOöLUa, Burçe ATAÇ MOGOLa, Vural GÖKMENa,b

a
Department of Food Engineering, bFood Research Center, Hacettepe University, Ankara, Turkey
(neslihangoncuoglu@hacettepe.edu.tr, burcea@hacettepe.edu.tr, vgokmen@hacettepe.edu.tr )

INTRODUCTION
Deep-fat frying is one of the most widely used process in the world. During deep-fat frying
frying, oils are exposed to high temperatures ~160-180°C and when they come into contact
with frying material several chemical reactions occur. Two of the most important reactions
occurring during frying are the Maillard reaction and lipid oxidation in food and oil,
respectively. These reactions generate neo-formed compounds that are potentially harmful
from the viewpoint of health. These harmful compounds, so called thermal process
contaminants accumulate in frying oil during its repetitive use for practical reasons. It is
undesirable to use highly contaminated frying oil because it becomes a significant part of the
fried product. The worldwide production of edible vegetable oil is about 60 million tonne a
year [1], most of this oil is used for frying and being discarded. Discarding frying oils
immediately after use is not reasonable because of economical and waste disposal problems
which endangers environment. Regenerating used frying oils by adsorption as a solution not
only reduce disposal problems, but, more importantly, would increase the quality of oil.

MATERIALS & METHODS

Intensively used frying oil containing reasonably high concentration of HMF (~6 mg/L) was
used. The suitability of six styrene-divinylbenzene based adsorbent resins for removal of HMF
from oil was tested. The successive use of resin was also tested after reconditioning with
hexane. Among resins one was selected, whose oil regeneration ability is higher, for further
analysis. Adsorption experiments were performed by mixing 100 mL of oil with different
amounts of resin (1.0, 2.5, 5.0, 7.5, 10.0 g) at different temperatures (30, 40, 50°C).

RESULTS & DISCUSSION

The objective of this study was regeneration of frying oils that contain high amount of
Hydroxymethylfurfural (HMF) as a thermal process contaminant. In order to monitor the
success of the regeneration process, it is advantageous to choose a marker molecule indicating
the level of frying oil contamination, rather than a global index like total polar compounds.
HMF is formed during the Maillard reaction [2] and sugar decomposition [3] reactions at high
temperatures. Due to its partial polar structure, it may transfer to oil from food during frying.
The results revealed that styrene-divinylbenzene based resins are capable of removing HMF
from oil. Of the resins tested, Dowex SBR LC NG(OH) provided 99.3% reduction in HMF
concentration of oil within 1.5 hrs (Figure 1).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1825
 Figure 1. HMF reduction of styrene-divinylbenzene based resin, Dowex SBR LC NG(OH) during
adsorption process

Equilibrium adsorption data was fitted to different adsorption models such as Langmuir,
Freundlich etc. to describe the adsorption behavior of HMF onto resin. The data fitted well to
Langmuir model at all temperatures studied. The adsorption process followed pseudo-second
order kinetics, and pore diffusion was found to be the effective adsorption mechanism. The
resin was able to remove ~90% of HMF in oil even after fifth adsorption cycle.

CONCLUSION
HMF should be taken into consideration as a quality index of frying oils. Dowex SBR LC
NG(OH) , styrene-divinylbenzene based resin, was found to be an effective solution to remove
HMF from highly contaminated frying oil.

REFERENCES
[1] United States Department of Agriculture 2000. United States Government Printing Office,
Washington, USA. In Agricultural Statistics pp. III-26.
[2] Ames, J. M. 1992. Biochemistry of Food Proteins. In : B. J. F. Hudson (Eds). Elsevier Applied
Science London pp. 99, 153.
[3] Kroh, L. W. 1994. Caramelisation in food and beverages. Food Chemistry, 51, 373-379.

1826
 Extending shelf life of watercress by means of alternative sanitizers and modified

atmosphere packaging
a,b a a a,c

a
Cielo Char , Paulina Villena , Andrea Hinojosa and Víctor Escalona
b
Center of Postharvest Studies, University of Chile. Santiago, Chile. web site: www.cepoc.cl.
Agroindustry and Enology Department, Fac. Agricultural Sciences, University of Chile, Santiago, Chile
c
(cdchar@u.uchile.cl)
Agricultural Production Department, Fac. Agricultural Sciences, University of Chile, Santiago, Chile
(vescalona@uchile.cl)

INTRODUCTION
Manipulation of vegetables and fruits causes physiological stress and wounds, resulting in
increased respiration rate and ethylene production, membrane deterioration, water loss and
susceptibility to microbial contamination [1]. The disinfection is one of the most critical steps
in the development of minimally processed fresh (MPF) products affecting the quality, safety
and shelf-life of the end product [2]. The aim of this research was to extend shelf life of ready
to eat watercress salads by means of the reduction of the respiration rate and prevention of
spoilage proliferation. The combination of passive modified atmosphere with peroxyacetic acid
(PAA), chlorine dioxide (CD) and acidified sodium chlorite (ASC) were assessed. Headspace
gases evolution and native flora were monitored at selected time intervals during 12 days of
refrigerated storage.

MATERIALS & METHODS

Watercress leaves were pre-washed with water at 5º C for 5 min to remove any foreign
material. Leaves
-1 were immersed for 3 min in different sanitizer
-1 solutions: chlorine dioxide (5
or 10-1mg L ), acidified sodium chlorite (250 or 500 mg L ) and peroxyacetic acid (50 or 90
mg
-1 L ). The efficiency of the sanitizers was compared to that of sodium hypochlorite (100 mg
L ). Leaves
-2 -1 were packed (50 g) in -2plastic
-1 bags PD-961EZ of 28 x 13 cm, (permeability: 7000
mL m d O2 and 21,000 mL m d CO2), which were heat sealed generating a passive
modified atmosphere and stored at 5°C for 12 days. The evolution of O2 and CO2 inside the
plastic bags was monitored by taking gas samples with a 10 mL syringe and injected into a gas
chromatograph (Hewlett Packard 5890 Series II). For microbiological analysis 10 g were
placed in sterile bags with peptone water and processed in an stomacher. Serial dilutions were
plated to determine mesophilic aerobic bacteria (Plate Count Agar, 37°C 2d.);
Enterobacteriaceae (Eosine Methylene Blue, 37°C 2 d.); acido lactic bacteria (Man Rogosa
and Sharpe, 37°C 3 d); yeasts and molds (Acidified Potato Dextrosa Agar, 22°C 5 d) and
psychrophilic bacteria (Plate Count Agar, 5°C 7 d.). Three samples per treatment were
analyzed on two different runs.

RESULTS & DISCUSSION

-1 -1
Initial respiration rate values were in a range of 89 – 135 mg CO2 kg h and declined
-1 -1 during
storage for all treatments, reaching equilibrium after § 8 days (45 to 65 mg CO2 kg h ). These

11th International Congress on Engineering and Food - Athens, Greece, 2011 1827
high respiration rates could be explained by the wounding produced during the preparation
process and subsequent -1physiological response in the tissue. Initial lowest respiration rate was
obtained for 10 mg L CD treatment. Even though, after equilibrium was reached non
significant differences (p < 0.05) were observed among all treatments. -1 -1 In the second
experiment, initial respiration rates were lower (80-99 mg CO2 kg h ). However, during
storage respiration rates reached similar values. Oxygen levels decreased and CO2 levels
increased in all MA packages during the storage period at 5 ºC due to watercress respiration.
Oxygen level was reduced from § 18 % to a range of 5.7 - 2.2 % after 8 days of storage.
Carbon dioxide levels promptly increased during the first 24 h from less than 1% to more than
2.5%, and reached equilibrium levels § 3.5 % during storage. Only slight differences were
observed with the different treatments after 5-8 days of storage and non significant differences
(p < 0.05) were observed at the end of storage. In the second experience a similar pattern was
observed. Initial microbial load reduction in less contaminated raw material was greater for DC
and
-1 SH (1-1.2 Log units reduction of mesophilic bacteria) maintaing low counts (§ 5 logCFU
g ) during 13 days of storage. Higher reductions were obtained using raw material with higher
microbial load (1.9, 1.6 and 1.4 Log -1units for CD, ASC and SH, respectively). In spite of this,
all sanitizers exceeded 8 logCFUg at the end of storage. Modified atmosphere had an
inhibitory effect on microbial growth obtaining lower counts for SH with MA than in a
perforated bag. The different sanitizers reduced initial psychrotrophic bacteria counts (1.7- 1.5
Log units).
-1 SH was the most effective; however, DC ended up with the-1lowest counts (§ 4 log
CFU g ) at the end of storage. In cleaner raw material, CD (10 mg L ) and SH successfully-1
inhibited Enterobacteriaceae during -1 all storage accomplishing
-1 less than 5 Log CFUg . Higher
concentrations of PAA (90 mg L ) and ASC (500 mg L ) also achieved reasonable levels.
Conversely, when raw material came with high initial Enterobacteriaceae count, none of the
sanitizers was efficient enough to diminish that load to acceptable levels. Sanitizers reduced
mold and yeasts (0.5-0.85 log units). These microorganisms,-1as well as lactic acid bacteria
slightly increased during storage, but never exceed 2 logCFU g with none of the treatments.

CONCLUSION
All sanitizers and MA helped to maintain acceptable levels of microbial flora and reduced
respiration rate of watercress for 13 or 8 days-1depending on the sanitizer and the initial
microbial load of the raw material. DC (10 mg L ) would be a good alternative to replace SH
reducing the rates of deterioration and extending shelf life. It diminished initial microbial load
and also maintained low counts of all microorganisms during storage in the raw watercress
with low initial load. PAA and ASC also achieved reasonable counts at 8 days -1 of storage.
When the raw material had high initial load, all sanitizers exceeded 8 logCFU g at the end of
storage.

REFERENCES
[1] Rico D., Martín-Diana A.B., Barat J.M. & Barry-Ryan C. 2007. Extending and Measuring the Quality
of Fresh-Cut Fruit and Vegetables: a Review. Trends in Food Science and Technology, 18, 373-386.
[2] Gil M.I., Selma M.V., López-Gálvez F. & Allende A. 2009. Fresh-Cut Product Sanitation and Wash
Water Disinfection: Problems and Solutions. International Journal of Food Microbiology 134: 37-45.

1828
Modeling the effect of acid and osmotic shifts above and across the growth boundaries on

the adaptation and growth of Listeria monocytogenes

Charalampia-Irini A. Belessia, Sofia I. Merkouria, Antonia S. Gounadakia, Sol Schvartzmanb, Kieran

Jordanb, Eleftherios H. Drosinosa and Panagiotis N. Skandamisa

a
Laboratory of Food Quality Control and Hygiene, Department of Food Science and Technology,
Agricultural University of Athens, Greece. Correspondence: pskan@aua.gr
b
Teagasc, Dairy Products Research Centre, Moorepark, Fermoy, Co. Cork, Dublin

INTRODUCTION
Exposure of bacteria to dynamic environmental conditions may pose additional adaptation
work, expressed in the form of intermediate lag time [3, 5]. Acid and osmotic shifts pose a
higher energetic burden than temperature shifts, especially around the growth boundaries. In
the present study, Listeria monocytogenes growth was evaluated at 10°C, being shifted after (i)
growth at growth permitting pH and aw levels or (ii) habituation at no-growth acid and osmotic
conditions for up to 10 days.

MATERIALS & METHODS

A L. monocytogenes isolate (102 cfu/ml), which persisted for 10 years in the environment of a
dairy plant was grown to late-exponential phase in TSBYE at 7 pH (7.2, 6.5, 6.0, 5.8, 5.5, 5.3,
5.1 at aw 0.995), adjusted with lactic acid and 5 aw (0.995, 0.97, 0.95 and 0.93 at pH 7.2).
When L. monocytogenes reached ca. 8-9 log cfu/ml in each of the above conditions, it was
shifted to all the remaining growth-permitting pH and aw levels at 10°C. Shifts from growth to
no growth conditions were also carried out by transferring L. monocytogenes, habituated at pH
4.9 and aw 0.90 for 1, 5 and 10 days to all the growth permitting conditions. Growth curves
were fitted with the Baranyi model [1]. Secondary models based on multiplicative effects of
each factor as well as an interaction term (ȟ) were developed to predict lag time and ȝmax in
response to pH and/or aw shifts also taking into account pH and aw before each shift. A similar
approach was adopted for the work-to-be done (ho) of cells following each shift.

RESULTS & DISCUSSION

Acidic and osmotic downshifts linearly decreased the maximum specific growth rate of L.
monocytogenes. The lag time of the organism increased with all osmotic downshifts, as well as
with the reduction of pH to 5.1. Conversely, any type of shift within pH 5.5-7.2 did not
markedly affect the lag times of L. monocytogenes. Downshifts to pH 5.1 induced 96 and 142 h
of lag to cultures derived from pH 7.2 and 5.3, respectively [3], whereas cultures grown at pH
5.5-6.0 showed no lag time. With regards to the habituation at no-growth conditions, longer
incubation of the cells at aw 0.90, increased their subsequent growth initiation, suggesting
adaptation to osmotic stress. Conversely, extended habituation at pH 4.9 reduced subsequent
growth of L. monocytogenes, possibly due to cell injury. These results suggest that there is an
adaptation or injury rate induced at conditions inhibiting the growth of the pathogen (Fig. 1a).
A dynamic model, describing the effect of both the magnitude of shifts and the initial pH or aw,
successfully predicted growth of L. monocytogenes in milk and cheese during ripening (pH

11th International Congress on Engineering and Food - Athens, Greece, 2011 1829
6.4-5.5 and aw 0.98 to <0.90; Fig 1b). In contrast, using ȝmax as a function of momentary pH
and aw and/or assuming that no adaptation work is induced due to environmental shifts resulted
in significant overestimation of L. monocytogenes growth, especially during downshifts in pH
and aw. Contrary to the observations with aw, the longer the cells were held at no growth pH
(4.9), the higher was the work needed for growth initiation in a more favourable environment.
This may be explained on the basis of the higher energetic burden posed by pH on cells
compared to aw at no growth conditions [4].

Figure 1: Conceptual basis of the model for the lag time of L. monocytogenes (Į) in response to aw or pH
shifts back and forth across the growth boundary and growth of L. monocytogenes (Ŷ,b) at 10°C in milk
under simultaneously varying conditions of pH (dotted and dashed line) and aw (dotted line), shown as a
comparison between the model predictions with (continuous line) and without (dashed line) taking into
account the effects of sudden shifts (From: Le Marc et al., [2]).

CONCLUSION
A dynamic model capable of predicting growth of L. monocytogenes in response to pH and aw
shifts, needs to consider: (i) the dependence of μmax on the magnitude and the directions of pH
and aw shifts; (ii) the increase of h0 in case of osmotic or pH downshifts; and, (iii) the rate of
adaptation work performed during habituation at no growth aw and the ‘rate of injury’ of cells
habituated at no growth pH.

REFERENCES
[1] Baranyi, J. & Roberts, T. A. 1994. A dynamic approach to predicting bacterial growth in food.
International Journal of Food Microbiology 23, 277-294. [2]Le Marc Y., Skandamis P.N., Belessi C.I.A.,
Merkouri S.I., George S.M., Gounadaki A.S., Schvartzman S., Jordan K., Drosinos E.H., & J. Baranyi.
2010. Modeling the Effect of Abrupt Acid and Osmotic Shifts within the Growth Region and across
Growth Boundaries on Adaptation and Growth of Listeria monocytogenes, Applied and Environmental
Microbiology, 76(19), 6555–6563. [3] Robinson T.P., Ocio M.J., Kaloti A. & Mackey B.M. 1998. The
effect of the growth environment on the lag phase of Listeria monocytogenes. International Journal of
Food Microbiology 44, 83-92. [4] Shabala L., Lee S.H., Cannesson P. & Ross, T. 2008. Acid and NaCl
limits of growth of Listeria monocytogenes and influence of sequence of inimical acid and NaCl levels on
inactivation kinetics. Journal of Food Protection 71, 1169-1177. [5] Swinnen I.A.M., Bernaerts K.,
Gysemans K., & Van Impe J.F. 2005. Quantifying microbial lag phenomena due to a sudden rise in
temperature: a systematic macroscopic study. International Journal of Food Microbiology 100, 85-96.

1830
Effect of contamination stage and inoculum history on the survival and growth of Listeria

monocytogenes in semi-hard and hard cheese

Charalambia-Irini A. Belessi, Sonia Arapaki, Antonia S. Gounadaki and Panagiotis N. Skandamis

Laboratory of Food Quality Control and Hygiene, Department of Food Science and Technology,
Agricultural University of Athens, Greece. Correspondence: pskan@aua.gr

INTRODUCTION
Listeria monocytogenes is a food borne pathogen of major concern for the dairy industry. Dairy
products are commonly ready-to-eat products and the evaluation of their ability to support
growth or survival of this pathogen is crucial for risk assessment. Previous studies have
demonstrated L. monocytogenes behavior in soft whey cheese [5], in semi-hard [4] and hard
ripened cheese [2], in most cases, when contamination takes place at the final product.
Nevertheless, contamination of a product in a dairy factory can occur at various stages (i.e.,
storage, production and ripening) and from different sources (i.e. unpasteurized milk, final
products, humans, surfaces, insects). All sources provide cells with different physiology and
probably different capacity to overcome the hurdles posed to L. monocytogenes throughout
manufacture and ripening of dairy products, as well as by their physicochemical properties.
The objective of the present study was to determine L. monocytogenes survival or inactivation,
after habituation or biofilm formation at various environments, in a semi-hard (Feta) or a hard
(Graviera) cheese, contaminated at different stages of manufacture procedure.

MATERIALS & METHODS

Planktonically grown and detached L. monocytogenes cells were prepared as inocula (twelve in
total), when seven L. monocytogenes isolates (from cheese, dairy surfaces or humans) were left
to grow separately in: TSBYE with 1% glucose (acid adapted) or without glucose
(nonadapted), Maximum Recovery Diluent (MRD), full fat milk, and commercial Feta and
Graviera cheese in the presence of stainless steel coupons (2x5cm2) for 3 days at 20°C and then
detached by bead vortexing. The inocula were used separately to inoculate (102 CFU/g or ml)
pasteurized milk, curd during cutting, Feta and Graviera cheese after ripening. At all cases full
manufacture was followed, including ripening and storage at 4oC.

RESULTS & DISCUSSION

Since cross contamination with L. monocytogenes can occur at various parts of cheese
manufacturing it is important to investigate its behavior after contamination at each part
separately. The three scenarios chosen were of the major critical control points of Graviera and
Feta manufacturing procedures [1, 3]. Inoculation after milk pasteurization resulted in growth
of all inocula at both cheeses (Fig 1a, b). Most of inocula types remained above regulatory
levels (100 cfu/g) during 90 days of storage in both Graviera and Feta cheeses (Fig 1a, b).
Inoculation during curd cutting resulted in a long-term survival on Graviera cheese (possibly
due to the reheating stage after cutting in Graviera cheese technology), while in Feta cheese, a
significant growth was followed by inactivation, in most cases. Initial growth was not observed

11th International Congress on Engineering and Food - Athens, Greece, 2011 1831
in Graviera cheese due to reheating of the product. When inoculated after ripening, L.
monocytogenes cells survived throughout storage (82 days) of Graviera cheese at low levels
(<2 log CFU/ml), while rapid reduction (<20 days) of all inocula was observed in Feta cheese.
Concerning inoculum history, TSBYE detached (BF) and milk planktonic (PL) cells
demonstrated higher growth, when inoculated in milk or curd, or higher survival (in cheeses)
than other inocula, suggesting that prior habituation in the ecosystems of Feta and Graviera
cheese or MRD probably poses large energetic burden to L. monocytogenes cells, rendering
them susceptible to subsequent stresses.

(a) Graviera cheese

(b) Feta cheese
8 8

7 7

6 6
log cfu/g or cm2

5 5
4 4
3 3
2 2
1 1
0 0
0 20 40 60 80 100 0 20 40 60 80 100
days days
TSBYE-PL TSBYE-BF Milk-PL Milk-BF Graviera-PL Graviera-BF TSBYE-PL TSBYE-BF Milk-PL Milk-BF Feta-PL Feta-BF

Figure 1. L. monocytogenes counts in Graviera (a) and Feta (b) cheese samples, during storage at 4oC.
Cheese samples were inoculated after milk pasteurization with L. monocytogenes cells previously
habituated (PL) or formed bioflms (BF) in TSBYE, milk, Graviera cheese or Feta cheese.

CONCLUSIONS
The results may address safety implications relevant to the diversity of L. monocytogenes cells
in a food processing environment and their variant ability to proliferate or survive in dairy
products, which might pose risks to public health. Furthermore, the importance of the stage
contamination is highlighted, as if contamination occurs at the first stages of the manufacturing
procedure, L. monocytogenes can proliferate and the final physicochemical properties of the
dairy product may not secure satisfactory inhibition of the pathogen.

REFERENCES
[1] Arvanitoyannis I.S. & Mavropoulos A.A., 1999. Implementation of the Hazard Analysis Critical
Control Point (HACCP) System to Kasseri/Kefalotiri and Anevato Cheese Production Lines. Food
Control, 11, 31-40. [2] Giannou E., Kakouri A., Matijasic B.B., Rogelj I., & Samelis J. 2009. Fate of
Listeria monocytogenes on Fully Ripened Greek Graviera Cheese Stored at 4, 12, or 25 0C in Air or
Vacuum Packages: In Situ PCR Detection of a Cocktail of Bacteriocins Potentially Contributing to
Pathogen Inhibition. Journal of Food Protection. 72, 531–538. [3] Mavropoulos A.A. & Arvanitoyannis I.
S., 1999. Implementation of Hazard Analysis Critical Control Coint to Feta and Manouri Cheese
Production Lines. Food Control 10, 213-219. [4] Papageorgiou D.K., & E.H. Marth, 1989. Fate of
Listeria monocytogenes During the Manufacture, Ripening and Storage of Feta Cheese. Journal of Food
Protection, 52, 82-87. [5] Papageorgiou D.K., M. Bori & A. Mantis, 1996. Growth of Listeria
monocytogenes in the Whey Cheeses Myzithra, Anthotyros, and Manouri During Storage at 5, 12, and
22°C. Journal of Food Protection 59:1193-1199.

1832
 Inoculated pack study of an intermediate moisture egg patty

Michelle Richardson a, Anthony Sikes, Claire Lee, and Sydney Walker

U.S. Army Natick Soldier Research Development & Engineering Center, Natick, MA, USA
(michelle.j.richardson@us.army.mil)

INTRODUCTION
The number of breakfast items containing eggs available to military personnel is very limited
and not highly acceptable. This limited variety of breakfast items results in noticeable
increases in menu monotony, decreased consumption, and reduced nutritional intake.
Therefore a need exist to provide military personnel with breakfast components that are
microbiologically stable and highly acceptable. Previous studies show sodium laureate (SL)
EDTA and butylated hydroxyanisole (BHA) alone and in combination with Nisin was effective
in preventing the growth of three strains Staphylococcus aureus in intermediate moisture egg
patties. However, when compared to other antimicrobials, Nisin is very expensive. Although
sucrose laurate is not as expensive as nisin, it is not readily available to commercial producers.
Thus objective of this study was to validate the use of an alternative antimicrobial ingredient
that is less expensive than Nisin and more readily available than sucrose laurate to inhibit the
growth of three strains of S. aureus in an intermediate moisture egg (IME) patty.

MATERIALS & METHODS

Dehydrated whole eggs (Henningsen, Omaha Nebreska); glycerol (KIC, Chemicals, Amonk,
NY); dehydrated egg white (Debel Foods Corp., Elizabeth NJ) nisin (Nisaplin, Danisco USA);
xantham gum (Keltrol F, Kelco, San Diego, CA); sodium acid sulfate (Jones Hamilto Co.,
Walbridge, OH); citric acid, sorbic acid, tartaric acid, and malic acid (Spectrum Chemical
Corp., Gardena, CA); glucono-į-lactone (Balchem Corp., New Hampton, NY); natural and
artificial butter flavor F004418 (Edlong Flavors, Elk Grove Villlage, IL); and 100cc oxygen
scavengers (Mutlisorb Technolgoies, Buffalo, NY). Bacteria used for inoculation were three
strains of Staphylococcus aureus obtained from the American Type Culture Collection (ATCC,
Rockville MD). The strains included ATCC 27154, ATCC 6538 and ATCC 8095. Media and
diluents used included tryptic soy agar (TSA, Dif co) supplemented with 0.5 percent yeast
extract, Tryptic soy broth (TSB, Difco); TSB-yeast extract (Difco); 0.1 percent sterile peptone
water (Difco); Butterfield ‘s Phosphate Buffer pH 7.2; and Staphylococcus 110 culture agar,
(DIFCO).
The experimental units consisted of four variables: 20 gram egg patties that were (1) untreated
positive control, with an average water activity (aw) of 0.989 and pH of 7.23; (2) treated
positive control with 3.5% L Sodium Lactate (Purasal S100) with an average aw 0.989 and pH
6.55; (3) untreated intermediate moisture control, with an average aw of 0.919 and pH of 5.61;
and (4) treated intermediate moisture control with 3.5% Purasal S 100, with an average aw of
0.902 and pH of 5.6. An inoculum cocktail of three strains of S. aureus was delivered across
the surface of the IME patty; the products were packaged in tri-laminated pouches with
oxygen-scavenging sachets and put into storage for 6 months at 25°C. Microbiological
enumerations were performed at 0, 7, 14, 21, 28, 42, 56, 84, 105, 133, and 161days of storage

11th International Congress on Engineering and Food - Athens, Greece, 2011 1833
RESULTS & DISCUSSION
IMF items are appropriate for field feeding because they directly feed a faster, lighter
mobilized military 1. Previous studies show Sodium Laureate (SL) EDTA and Butylated
Hydroxyanisole (BHA) alone and in combination with Nisin was effective in preventing the
growth of three strains s. aureus in intermediate moisture egg patties 2. Indications are that
both the untreated and treated positive control patties supported the growth of s. aureus. There
was more than a three log increase for the control patties after 7 days of storage (25o C); the S.
aureus population was greater than 106 CFU/g throughout the study for both control samples.
The level of S. aureus did not increase during storage for the untreated and treated IME patties.
After 14 days the s. aureus population was below the minimum detection level (<10 cfu/g) for
the IME patties. The Purasal had no effect on s. aureus growth.

CONCLUSION
Hurdle technology was exploited to develop IME patties for use alone or as an ingredient in a
multi-component breakfast entrée for military field feeding. The various hurdles employed
were aw, pH, oxygen removal, pasteurization and bacteriocin utilization. The positive controls
had a three log increase over inoculum level and populations greater than 106 CFU/g.
Although toxin testing was not conducted, toxin production is likely (> 106 cfu/g) and the
positive control patties require time/temperature control. The s. aureus growth was limited for
the treated and untreated IME patties throughout the study and time/temperature control is not
needed, i.e., it is considered microbiologically stable. The limited s. aureus growth was not
due to purasal, however, the ability to provide military personnel with breakfast components
that are microbiologically stable and highly acceptable is feasible.

REFERENCES
[1]. Taoukis, P. S. and Richardson, M. (2008) Principles of Intermediate-Moisture Foods and Related
Technology, in Water Activity in Foods: Fundamentals and Applications (eds G. V. Barbosa-
Cánovas, A. J. Fontana, S. J. Schmidt and T. P. Labuza), Blackwell Publishing Ltd, Oxford, UK.
doi: 10.1002/9780470376454.ch11
[2]. Richardson, M., Sikes, A. 2008. Development of an intermediate moisture egg (IME) using hurdle
technology. Institute of Food Technologists Annual Meeting.

1834
 HACCP implementation in public hospitals: a survey in Crete, Greece

E. KokkinakisĮ,b, A. Kokkinakia, G. Kyriakidisb, A. Markakib, G.A. Fragkiadakisb

a
Technological Education Institute (TEI) of Crete, Department of Commerce and Advertising, Ierapetra,
Crete, Greece (manoskokkinakis@yahoo.gr; katrinakok@yahoo.gr)
b
Technological Education Institute (TEI) of Crete, Department of Nutrition and Dietetics, Siteia, Crete,
Greece (gregkyriakidis@staff.teicrete.gr ; markaki@staff.teicrete.gr; fragkiadakis@staff.teicrete.gr)

INTRODUCTION
In the island of Crete, Greece, 7 major hospitals offer their services in conjunction with a
number of public health centres. Between 2004 and 2009, personnel of the Technological
Education Institute (TEI) of Crete, carried out food safety surveys in these hospitals. Our basic
aim was to evaluate the degree of compliance to food safety [1] and to investigate the
difficulties that exist during the HACCP implementation process, in hospital mass-catering
systems. In this paper we summarise our relevant experience concerning the actual problems
we observed on implementing HACCP in the hospitals of Crete.

MATERIALS & METHODS

The evaluation process was based in personnel interviewing; checklist screening of important
processes: menu design, suppliers evaluation, incoming materials control, storage, freezing and
refrigeration, food preparation, food distribution and general HACCP prerequisites application,
e.g. sanitation procedures, water hygiene, pest control, personnel hygiene etc; microbiological
analysis in food and water samples. Two of the hospitals we surveyed were following HACCP
methodology, two had just started to introduce the system and the remaining three were
planning HACCP introduction. The overall results of the above screening have only been
partially presented [1, 2].

RESULTS & DISCUSSION

Studies concerning the barriers of HACCP implementation in hospitals [8], have underlined the
lack of HACCP prerequisites in hospitals; the lack of in-house HACCP skills; the high cost;
the long time; the staff turn-over regulations within the hospital; the lack of
management/owner commitment; the poor ownership of externally designed HACCP plans etc.
Some potential problems were detected in our survey also and unfortunately actual food-safety
problems did occur in these hospitals in the same period [3, 4].

Based on our actual findings from the 7 hospitals, we present a specific key-elements approach,
in order to both implement and successfully operate HACCP systems in public hospital mass-
catering systems. This approach could potentially be valuable to public health institutions or
authorities, currently under the process of implementing HACCP or quality management
system, such as ISO 9001:2000. Our proposed approach is based on fourteen key elements
(Table 1). A “managerial effect” was observed to be of great importance for efficient HACCP

11th International Congress on Engineering and Food - Athens, Greece, 2011 1835
implementation in the hospitals. Without active managerial involvement a HACCP system is
almost impossible to operate successfully.

Table 1. Crucial elements for HACCP implementation in hospitals

No Description
1. Managerial commitment
2. Availability and enforcement of risk-informed regulations
3. Hospital registered-dieticians, food technologists, and hygienists’ involvement
4. Patients menu’s planning within the HACCP system
5. Prevention of malnutrition in patients
6. Hospitals kitchen and food-management personnel training
7. Integration of HACCP procedures with other hospital functions
8. Hygiene supervision by central and regional public health authorities
9. External food delivery/introduction into the hospital
10. Handling of minimally processed food
11. Regular inspections of kitchenware
12. Food storage conditions
13. Food remaining disposal
14. HACCP-certified suppliers’ availability

CONCLUSIONS
Based on our actual findings and experiences after a survey in Crete, Greece, we emphasize
some elements, crucial for implementing and successfully operating HACCP systems in
hospitals. The specific requirements of HACCP in the hospitals, as well as the benefits that can
offer to patients must be further investigated and presented to the health professionals involved,
as nutritionists and/or dieticians, food technologists, nurses, biologists etc. Cooperation is
necessary between hospital administrators, public-healthy authorities, universities and research
centres, in order to advance the quality of the services that the Greek hospitals offer.

REFERENCES
[1] Kokkinakis E. & Fragkiadakis G.A. 2007. HACCP Effect on Microbiological Quality of Minimally
Processed Vegetables: a Survey in Six Mass-Catering Establishments. International Journal of Food
Science and Technology, 42(1), 18-23.
[2] Food Standards Agency, FSA 2000. Code of Practice No.9: Food Hygiene Inspections (Second
Revision, October 2000), London, UK.
[3] “Patris” Newspaper, “ISO certification for Venizeleion Hospital” (in Greek), November the 26th
2005, (http://www.patris.gr/articles/74175?PHPSESSID=5djb1h53aukdqsm5koi8ncc8r7).
[4] “Patris” Newspaper, “Innocent for Salmonella in PAGNH” (in Greek), February the 1st
2010, (http://www.patris.gr/articles/173570?PHPSESSID=5djb1h53aukdqsm5koi8ncc8r7).

1836
 HACCP implementation in local food industry: a survey in Crete, Greece

E. KokkinakisĮ,b, A. Kokkinakia, G. Kyriakidisb, A. Markakib, G.A. Fragkiadakisb

a
Technological Education Institute (TEI) of Crete, Department of Commerce and Advertising, Ierapetra,
Crete, Greece (manoskokkinakis@yahoo.gr; katrinakok@yahoo.gr)
b
Technological Education Institute (TEI) of Crete, Department of Nutrition and Dietetics, Siteia, Crete,
Greece (gregkyriakidis@staff.teicrete.gr ; markaki@staff.teicrete.gr; fragkiadakis@staff.teicrete.gr)

INTRODUCTION
Small food-processing companies contribute substantially to the production, manufacture and
retail of food in the periphery of most countries. For Crete, a leading destination depending on
tourism and on the agro-food sector, the high quality/safety and prestige of the food offered in
the island is crucial, in order to preserve the international recognition on the brand name
“Crete” and on the “Cretan diet” healthy lifestyle. The aim of our survey/study was to evaluate
the actual changes to the microbiological quality of locally produced/packed food following the
implementation of HACCP systems in three different enterprises: a. an ice cream producing
factory, b. a company preparing pre-packed sandwiches, c. a water bottling company.
Emphasis was given to the processing steps, transportation, storage and retailing.

MATERIALS & METHODS

We applied mainly Association of the Official Analytical Chemists (AOAC), and International
Organization for Standardization (ISO) reference-methods, using the recommended pre-
enrichments and selective enrichments; in addition, other methods as i.e. Petrifilm [1, 2, 3].

RESULTS & DISCUSSION

The results of our survey show the extent of the positive effects that a Hazard Analysis Critical
Control Points (HACCP) system, introduced in an ice-cream factory, had on both the
microbiological quality of the final product as well as on the total quality/hygiene management.
Among others, two main hygiene problems concerning the presence of the pathogen
Staphylococcus aureus in the final product and the contamination of the mains-supply water
with Enterococcus faecalis were detected and corrected [1]. Further linkage of the HACCP
system introduced in the factory to quality management systems, such as ISO 9001:2000, can
be possibly proved to provide higher quality/hygiene standards, along with higher awareness of
the factories customers (i.e. ice cream retailers) [1].

Concerning the sandwiches producing plant, no special food-safety problem was detected,
possibly due to the establishment of a HACCP system in the plant. The majority of the
sandwiches were usually consumed within 1-3 days, however, indicator bacteria values cause
some concerns on possible dangers to human health and prove the necessity of monitoring the
food-storage conditions at the retailer’s level [2]. Since, due to the modern way of life, the
consumption of “ready to eat” food, as pre-packed sandwiches, increases, and further research
is needed on the logistics and retailing chain.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1837
Finally, concerning the water-bottling company, the results indicated the need to improve the
HACCP system, in order to continuously monitor the water quality of the supply source in the
plant, and to fully implement the correct storage conditions, hygiene procedures, and customer
training at supermarkets [3]. The monthly storage in the supermarkets did not seem to affect
the microbial safety of the bottled water, since all the samples tested were within safety
specifications. The contribution of plant’s HACCP system was valuable in terms of
transportation conditions of the bottled water, since the microbial quality of the final products
did not change during the 1st month of storage in supermarkets. A small increase in
Heterotrophic Plate Count (HPC) levels, during the 2nd and 4th month of storage, was within
the safety limits with all the tested samples safe for human consumption [3].

Both in the case of the ice-cream factory [1], as of course and in the water-bottling plant [3],
the quality of the water source were proved crucial. The responsibility on this water quality
concerns the local self-administration, since it is influenced by the potable water offered by
self-administration companies [1] as well as by environmental parameters that influence
spring-water quality [3]. In the first case [1] specific technical problems arise, as i.e. whether
the water must be chlorinated and what may be the effect of chloride in the final product. In the
second case [3], monitoring of animal husbandry activities and use of agricultural pesticides is
crucial in preserving the environment and the water. The application of good agricultural
practises protocols, as the AGRO 2-1 & 2-2 can reduce microbial hazards for consumers and
furthermore can establish practices in compliance to modern European requirements [4].

CONCLUSION
In all cases studied, HACCP implementation had a positive impact on company functioning,
personnel professionalism, and raw-materials quality-standards, while it was effective in
reducing average microbiological counts during food preparation. However, further attention is
required, in order to sustain and improve the quality perception of the local food-processing
industry. During the implementation of HACCP, this industry faces special technical problems
and challenges that require innovation and close cooperation with the local tertiary education
and research infrastructure. This cooperation, in the years of crisis, is a prerequisite not only for
preserving food-safety but also for developing future innovation applications.

REFERENCES
[1] Kokkinakis E.N., Fragkiadakis G.A., Ioakeimidi S.H., Giankoulof I.B. & Kokkinaki A.N. 2008.
Microbiological Quality of Ice-Cream before and after HACCP Implementation: A Factory Case
Study. Czech Journal of Food Science, 26(5), 383-391.
[2] Kokkinakis E.N., Fragkiadakis G.A., Kokkinaki A.N. & Lapidakis N.E., 201?, Microbiological
Quality of Pre-Packed Sandwiches at the Retailing Level. Acta Alimentaria (under revision).
[3] Kokkinakis E.N., Fragkiadakis G.A. & Kokkinaki A.N. 2008, Monitoring Microbiological Quality of
Bottled Water as Suggested by HACCP Methodology. Food Control, 19(10), 975-961.
[4] Kokkinakis E., Boskou G., Fragkiadakis G.A., Kokkinaki A. & Lapidakis N. 2007. Microbiological
Quality of Tomatoes and Peppers Produced under the Good Agricultural Practices Protocol AGRO 2-
1 & 2-2 in Crete, Greece. Food Control, 18(12), 1538-1546.

1838
 A simplified method for determination of the sour cassava starch expansion property
a
Maria Janete Angeloni Marcon, aDiego Jacob Kurtz, aMarcelo Maraschin, aValéria Reginatto, bIvo
Mottin Demiate, aEdna Regina Amante*

a
Department of Food Science and Technology, Agricultural Sciences Centre, Federal University of Santa
Catarina, Rodovia Admar Gonzaga, 1346, Itacorubi, Florianópolis, Santa Catarina, Brazil, CEP 88034001
Email: eamante@cca.ufsc.br
*Correspondent author
b
Department of Food Engineering, Ponta Grossa Estadual University, Ponta Grossa, Paraná, Brazil.

INTRODUCTION
Expansion is a natural characteristic of polvilho azedo, defined as the growth rate of the dough
during oven cooking, also referred to as expansion rate [1]. It is directly related to specific
volume, expressed in cm3.g-1. Based on results of several other published works [2], including
advanced techniques for analysis such as ATR-FTIR (Attenuated total reflectance-Fourier-
transform infrared spectroscopy), DSC, X-Ray diffraction, and others, this present work
intends to show variables of easy determination that could be suggested as an easy analysis to
predict the performance of polvilho azedo.

MATERIALS & METHODS

Every bakery of every bakery in the centre of Florianópolis, Brazil, were interviewed. All
samples were characterized for moisture content (method AOAC 921.10) [3]. Acid factor, pH,
expansion rate, swelling power, and specific volume were determined for all samples, and
intrinsic viscosity was determined for commercial native cassava starch (five samples) and
commercial polvilho azedo (three samples). Swelling power was evaluated at 90ºC, as
described by, with modifications [4]. The solubility was expressed in weight percentage and
the swelling power in weight gain. The determination of baking expansion capacity followed
the procedures proposed by the CERAT – Tropical Root Centre [5]. The formulation was
made with 50 g of sample with 40 mL boiling water for the production of the dough [6].
Specific volume of the dough balls after cooking was determined through the rape seed
displacement method and calculated as displacement/weight (cm3.g-1) [7]. The Intrinsic
viscosity [8] of the cassava starch samples was measured with a thermostatized viscometer
Schott, AVS 350, CT 52, at constant temperature 30oC, capillary diameter 0.63 mm; capillary
constant (K) of 0.01511 mm2.s-2.

RESULTS & DISCUSSION

Table 01 shows the value of co-relation coefficients for several parameters of the starch
quality. The results of the samples studied in this work show that the commercial polvilho
azedo with more desirable expansion property has pH values between 3.47 and 4.18, and acid
factor values between 3.17 and 4.36. These minimum and maximum values can be useful as
limits to predict the performance of polvilho azedo. The weight of the samples of specimen can
also be related with their expansion because the weight of the cooked product made with a
good polvilho azedo is lower than a polvilho azedo with low expansion performance.
Considering the method used in this work for determination of specific volume [7], we can find

11th International Congress on Engineering and Food - Athens, Greece, 2011 1839
a high amount of data, enough to establish a relation between the lost of weight sample of
specimen and the expansion rate. It is possible to predict that a good polvilho azedo must show
weight loss around 40 to 45 % after cooking. This evaluation is independent of pH or acid
factor determination, but it is advantageous in comparison to the traditional method used in
bakeries because it uses only a minimal quantity of sample, oven, and scale and also because it
reduces time of assay.

Table 1. Co-relation coefficient between expansion rate, and acid factor, pH, swelling power, specific
volume and specific viscosity for several samples of polvilho azedo with high and low expansion
performance and native cassava starch.
Sour cassava starch quality parameter Expansion rate(*) co-relation coefficient
r
Swelling power 0.52
Acid factor 0.81
pH 0.92
Intrinsic viscosity (**) 0.93
Specific volume 0.97
* Data from samples of native cassava starch (n=6) and sour cassava starch produced in laboratory (20) and
commercial trade marks (17). **Native cassava starch (n=6); commercial sour cassava starch (n=3).

CONCLUSION
Among several quality variables such as intrinsic viscosity, swelling power, specific volume,
pH, and acid factor, only the last two could contribute to predict performance of polvilho azedo
for products obtained through the traditional process, which takes long time for fermentation.
The inclusion of the percentage of weight loss during cooking can be a good variable, along
with pH and acid factor, to establish a prediction of the expansion performance of polvilho
azedo.

REFERENCES
[1] Mestres, C., Boungou, O., Akissoe & N., Zakhia, N. 2000. Comparison of the expansion ability of
fermented maize flour and cassava starch during baking. Journal of Science Food Agriculture 80, 665 –
672.
[2] Marcon, M.J.A., Kurtz D.J., Raguzzoni, J.C., Delgadillo, I., Maraschin, M., Soldi, V., Reginatto, V. &
Amante, E. R. 2009. Expansion Properties of Sour Cassava Starch (Polvilho Azedo): Variables Related to
its Practical Application in Bakery. Stärke/Starch 61, 716 – 726.
[3] Association of Official Analytical Chemists (AOAC). Official Methods of Analysis. 16 ed. Gaithersburg;
1999.
[4] Leach, H.W., McCowen, L.D. & Schoch, T.J. 1959. Structure of the starch granule. I. Swelling and
solubility patterns of various starches. Cereal Chemistry 36, 534 – 544.
[5] Maeda, K.C. & Cereda, M.P. 2001.Avaliação de duas metodologias de expansão ao forno do polvilho azedo.
Ciência e Tecnologia de Alimentos 21, 139 – 143.
[6] Pereira, J., Ciacco, C.F., Vilela, E.R. & Texeira, L.S. 1999. Féculas fermentadas na fabricação de biscoitos:
estudo de fontes alternativas. Ciência e Tecnologia de. Alimentos 19, 287 – 293.
[7] Cereda, M.P. 1983. Avaliação da qualidade de duas amostras de fécula fermentada de mandioca (polvilho
azedo). Boletim da Sociedade Brasileira de Ciência e Tecnologia de Alimentos 17, 305 – 320.
[8] Leach, H.W. 1963. Determination of intrinsic viscosity of starches. Cereal Chemistry 40, 593 – 600.

1840
 Influence of room temperature on food safety in refrigerated display cabinet

Laguerre O.a, Hoang M.a, Alvarez G.a, Flick D.b

a
Refrigeration Process Engineering, Cemagref, 92160 Antony, France
(onrawee.laguerre@cemagref.fr;hong-minh.hoang@cemagref.fr; graciela.alvarez@cemagref.fr)
b
AgroParisTech, 16 rue Claude Bernard, 75231 Paris Cedex 05, France (denis.flick@agroparistech.fr)

INTRODUCTION
A survey carried out by our team [1] showed that 30% of products presented in refrigerated
display cabinet were subjected to temperature abuse (more than 2°C higher than recommended
preservation temperature). Willocx et al [2] carried out a survey on processed vegetables in
Belgian retail display cabinets. This study also showed that retail display cabinets are a critical
point in the cold chain. Evans et al [3] observed that in open front display cabinet, the majority
of high temperature packs (97%) were located at the front and the largest number (60%) of
them was at the front base.
This work was carried out to; firstly, experimentally study the influence of the room
temperature on the product temperature in an open refrigerated display cabinet. Then, these
product temperatures were used in a predictive microbiological model to estimate the growth
of Listeria monocytogenes.

MATERIALS & METHODS

Figure 1 shows the side view of the display cabinet used in our study which was equipped with
one air curtain and 5 shelves. It was loaded with packages of test product made of
methylcellulose. Some packages were instrumented by calibrated thermocouples (T-type).
The display cabinet was located in a test room in which the room temperature was controlled at
20, 25 and 30°C. The temperature of air and test packages was measured every minute until the
steady state was reached.

RESULTS & DISCUSSION

The average temperature was calculated over 3h of the steady state period and reported in
figure1. The rise of room temperature leads to increase the air and the load temperatures
particularly at the front of the display cabinet.
A simple predictive model was used assuming a first order growth rate. The growth of Listeria
monocytogenes was estimated at various product temperatures after 4 days storage (Table 1).

CONCLUSION
Higher room temperature leads to higher air and product temperatures particularly the one
located at the front. This can be explained by the external air infiltration and the heat loss from
the display cabinet. The product temperature was used in a predictive microbiological model to
estimate the Listeria monocytogenes growth. This approach can be used as a tool of risk
evaluation.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1841
 a-Troom = 20°C b-Troom = 25°C c-Troom = 30°C

Figure 1. Product and air temperatures in the studied display cabinet.

bold and italic=air temperature, underlined= surface temperature of package, not underlined = centre
temperature of package

Table 1. Influence of product temperature on the growth of Listeria monocytogenes after 4 days storage.
Temperature Log[N(t)/N0)]
1.3°C (lowest observed product temperature) 0.13
4.0°C (maximal recommended storage temperature) 1.2
7.0°C 3.8
9.9°C (highest observed product temperature) 7.4

REFERENCES
[1] Cemagref & ANIA 2004. La chaine du froid du fabricant au consommateur: résultats de l'audit
ANIA/Cemagref. Revue Générale du Froid, 1042, 29-36.
[2] Willocx, F., Hendrick, M., Tobback, P., 1994. A preliminary survey into the temperature conditions
and residence time distribution of minimally processed MAP vegetables in Belgian retail display
cabinets. International Journal of Refrigeration, 17(7), 436-444.
[3] Evans, J.A., Scarcelli S. & Swain M.V.L. 2007. Temperature and energy performance of refrigerated
retail display and commercial catering cabinets under test conditions. International Journal of
Refrigeration, 30, 398-408.

1842
Antemortem and postmortem biochemistry, drip loss and lipid oxidation of European sea

bass muscle tissue

Nathanailides Cosmas, Panopoulos Soctrates., Kakali Fotini, Karipoglou Costas, Lenas Dimitrios

Dept Aquaculture & Fisheries, TEI of Epirus, Igoumenitsa, Greece,(cosmasfax@yahoo.com)

INTRODUCTION
Slaughtering methods of farmed fish can influence swimming activity and stress levels with
consequences for the flesh quality properties of fish fillets. For example, muscle metabolic
activity prior to death can affect the concentration of oxygen, ATP and the pH of muscle.
Handling stress of fish prior to harvesting is characterized by increased swimming activity
which leads to anaerobic metabolism while energy stores are depleted and white muscle pH
drops leading to acceleration of autolytic reactions after death with consequences for the
organoleptic parameters of fish fillets and the filleting yield of farmed fish[1]. In addition to the
food quality aspects of the slaughtering method, the potential suffering of fish being handled
during common aquaculture procedures or during slaughtering prompted scientific research in
the welfare of farmed fish during harvesting [5]. The purpose of this work was to investigate
the effect of handing stress on post- mortem muscle pH and rigor mortis of sea bass.

MATERIALS & METHODS

European sea bass were harvested using two different levels of handling stress. In one tank
(High handling stress group, HHSG) the water was lowered and the fish were captured using a
net and the fish were killed by immersion in and ice cold bath (4 part ice: 1 part sea water). In
the other tank (Lower handling stress group, LHSG) the level of water was lowered and fish
were anaesthetized moderately by immersion in a 30mg l-1 clove oil bath for 5 minutes.
Subsequently the fish were slaughtered by immersion in ice cold sea water (4 part ice:1part sea
water). After 30 min, the fish were stored with the ventral side upwards in ice. After 30 min,
the fish were stored with the ventral side upwards in ice. Body temperature of the fish was
determined at regular intervals using a temperature probe. Muscle pH was monitored shortly
after death and after ice storage. Drip loss, and thiobarbituric acid reactive substances
(TBARS) were measured in ice stored fillets.

RESULTS & DISCUSSION

The different methods of slaughtering used in the present work varied in terms of the
anticipated level and duration of stress. The fish of the HHSG exhibited significantly longer
period of escape swimming activity during the immersion in an ice cold bath. Moderate
anaesthetised fish (LHSG) exhibited minimal and short duration swimming activity during the
immersion in the ice slurry. The levels of lactate present in the muscle tissue of the two groups
varied according to the killing method, the lactate levels of the HHSG fish were higher
(11.64±1.08ȝmoles g-1) than the LHSG fish (9.6±1.1ȝmoles g-1). This difference in lactate
levels indicates a higher level of slaughtering stress in the HHSG. Under handling stress, fish
exhibit an escape swimming behaviour, which utilises the anaerobic metabolic pathways of the
fast white skeletal muscle.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1843
Table 1. Fillet pH drip loss and levels of Thiobarbituric acid reactive substances (TBARs) after 12 days
ice storage of sea bass. LHSG: Clove oil treated fish; HHSG: High handling stress group. n=10 per
group, Average values (+/- s.d.).
LHSG(Clove oil treated fish) HHSG (Control) ANOVA
pH 6.59 (0.06) 6.58 (0.08) NS,(P=0.7)
Fillet Drip loss (%) 4.77 (0.48) 5.39 (0.52) P=0.006
TBARS 1.04 (0.07) 1.16 (0.02) *
(ȝg MDA mg–1) P=0,001)

The results indicate that the intensity of handling stress during slaughtering of sea bass can
influence the development of biochemical changes and the onset of rigor mortis. Our results
support the hypothesis that increased levels of handling stress can lead to increased lipid
oxidation in the fillets of the harvested fish [2]. The handling stress prior to slaughtering results
in rapid ATP depletion and reduced nucleotides such as NAD+ and NADP+ which are
involved in the regeneration of various pro-oxidant substances [3] may have contributed in the
difference of lipid oxidation between the stressed group (HHSG) and the clove oil treated fish.
Lipid oxidation of fish fillets is highly undesirable with negative consequences for the
organoleptic and the nutritional parameters of the fillets. The clove oil treated fish exhibited
lower rate of lipid oxidation and drip loss during storage. In agreement with previously works,
on sea bass and sea bream [1] the result indicate that a reduction of slaughtering stress can
reduce the suffering of farmed fish during harvesting and also improve the quality of the fillets
[4,5] during storage, with benefits for both the animals and the consumers.

CONCLUSION
Our results indicate the significance of slaughtering procedure for both the food quality aspects
and the welfare of the harvested farmed fish. In the commonly used ice-slurry slaughtering
method, the death of the un-anaesthetized fish of the HHSG appeared to progress much slower
than the clove oil treated fish. This long agony of the HHSG appears to be a less humane
killing method with consequences for the meat quality of the harvested fish.

REFERENCES
[1] Bagni M., Civitareale C., Priori A., Ballerini A., Finoia M., Brambilla G., Marino G. 2007. Pre-
slaughter crowding stress and killing procedures affecting quality and welfare in sea bass
(Dicentrarchus labrax) and sea bream (Sparus aurata). Aquaculture 263, 52–60.
[2] Giuffrida A., Pennisi L., Ziino G., Fortino L., Valvo G., Marino S., Panebianco A. 2007. Influence of
slaughtering method on some aspects of quality of gilthead seabream and smoked rainbow trout.
Veterinary Research Communications, 31(4), 437-446.
[3] Hultin H.O., 1992. Biochemical deterioration of fish muscle. In: Huss, H.H. Jakobsen M. and Liston,
J. (eds.), Quality Assurance in the Fish Industry (Elsevier, Amsterdam), 125–138.
[4] Ribas L., Flos R., Reig L., MacKenzie S., Barton B.A., Tort L., 2007. Comparison of methods for
anaesthetizing Senegal sole (Solea senegalensis) before slaughter: stress responses and ¿nal product
quality. Aquaculture 269, 250–258
[5] Sigholt T., Erikson U., Rustad T., Johansen S., Nordtvedt T., Seland A., 1997. Handling stress and
the temperature affect meat quality of farmed-raised Atlantic salmon Journal Food Science 62, 868–
872.

1844
 Impact of initial handling and subsequent storage conditions on the safety and keeping

quality of sardines

K. Chatzikyriakidoua,b and E. Katsanidisa

a
Department of Food Science and Technology, Faculty of Agriculture, Aristotle University of
Thessaloniki, Box 235, GR-54124 Thessaloniki, Greece. (e-mail: ekatsani@agro.auth.gr)

b
Current affiliation: Department of Food Science, College of Agriculture and Life Sciences, University of
Wisconsin, Madison, WI, USA. (e-mail: hatzikyriakidouk@yahoo.gr)

INTRODUCTION
Sardine (Sardina pilchardus) is a highly perishable food commodity with a very short shelf-
life. Biogenic amines (including histamine, putrescine and cadaverine) are formed due to
microbial activity in fish tissue and they can cause adverse health effects for the consumers.
Total volatile basic nitrogen (TVBN) and oxidation levels (as measured by the TBA test) are
indicators of the safety and quality of the fish. It is hypothesized that varying initial handling
conditions, which may not cause immediately detectable changes, can have a very significant
impact on the safety and quality of fish during subsequent storage.
The objective was to study and model the impact of the initial handling conditions (during the
first 24 hrs) on the safety and quality of sardines subsequently stored at different temperatures.

MATERIALS & METHODS

Freshly caught sardines were initially stored at 0, 5 and 10 °C for 24 hrs in high precision (±0.2
°C) low temperature incubators. Then, they were vacuum-packaged and stored at either 4 or 8
°C for 12 and 5 d, respectively. The pH, total volatile basic nitrogen (TVBN, official EU
method 95/149/ǼU), oxidation levels (using the TBA test [1]) and the concentration of
putrescine, cadaverine and histamine (using HPLC analysis [2]) in the fish tissue were
measured throughout the storage period. Specific reaction rates for fish exposed to different
initial conditions were calculated where appropriate.

RESULTS & DISCUSSION

No significant differences were observed in the pH and TBA values of sardines initially stored
at 0, 5 and 10 °C. A small increase in pH during storage can be attributed to proteolysis and
decarboxylation reactions. The TBA values for samples initially stored at 5 and 10 °C reached
their maximum value sooner than the samples maintained at 0 °C. The TVBN increased over
time and samples initially stored at higher temperatures exhibited a faster development of
TVBN, for both storage temperatures (Table 1). Significant differences in the concentrations of
biogenic amines in the fish tissue were observed throughout storage at 4 and 8 °C, with the 8
°C samples reaching higher levels of biogenic amines (Table 1). Samples exposed to lower
initial temperatures exhibited a slower rate of biogenic amines production during storage at

11th International Congress on Engineering and Food - Athens, Greece, 2011 1845
both temperatures. The rates of putrescine, cadaverine and histamine formation were calculated
for the different initial and subsequent storage temperature conditions.

Table 1. Effect of initial handling conditions on the TVBN and histamine levels of sardines stored at 4
and 8 °C *
Stored at 4 °C Stored at 8 °C
Initial TVBN (%mg) Histamine (ppm) TVBN (%mg) Histamine (ppm)
conditions
0 °C 29.7a 21.0a 20.3a 7.1a
b b b
5 °C 39.6 130.5 27.8 125.2b
c b c
10 °C 41.9 150.6 33.3 208.8c
* Values are least square means for the duration of the storage period (12d at 4 °C and 5d at 8 °C).
a,b,c
Different superscripts in the same column denote statistically significant differences.

CONCLUSION
The initial handling temperature (during the first 24 hrs after the fish are caught) is a
determining factor for the safety and quality of sardine. Regardless of the subsequent storage
temperatures, the biogenic amines formation is accelerated significantly if the sardines are
initially exposed to elevated temperatures (5 or 10 °C). A small temperature abuse during the
distribution chain may not be immediately detected, but it can affect the safety and keeping
quality of fish.

REFERENCES
[1] Salih, A.M., Smith, D.M., Price, J.F. and Dawson, L.E. 1987. Modified extraction 2-thiobarbituric
acid method for measuring lipid oxidation in poultry. Poultry Sci. 66:1483-1488.
[2] Malle P. & Vallé M. 1996. Assay of biogenic amines involved in fish decomposition. Journal of
AOAC International, 79(1): 43-49.

1846
 Survival of Salmonella and Escherichia coli O157:H7 during freezing, thawing and

cooking of ground beef patties, simulating common household practises

Stavros G. Maniosa, Thomas Giovanisa, Argiro Lalechoua, Panagiotis N. Skandamisa

a
Laboratory of Food QualityControl and Hygiene, Department of Food Science and Technology,
Agricultural University of Athens, Iera Odos 75, 118 55, Athens, Greece. (pskan@aua.gr)

INTRODUCTION
Ground beef patties constitute conventional Ready-To-Cook meat products, ranked among the
top most frequently consumed meat products worldwide. These products may be prepared in-
house from ground beef or may also be found available in the form of pre-shaped frozen
patties, which is highly convenient for Quick Service Restaurants. In the recent years, ground
beef has been linked with several outbreaks of foodborne diseases caused by Salmonella or
Escherichia coli O157:H7. Therefore, Food Safety authorities have issued general guidelines
for the proper handling of ground beef in households or catering services. Through these
guidelines it is recommended that ground beef should be stored in refrigerators for 1-2 days or
in freezer up to 4 months, thawed in refrigerator or in microwave and cooked until the internal
temperature of patties reaches 71oC. However, the storage and cooking practices being applied
by the consumers, either in households or in catering services and restaurants, are generally
based on personal preferences and convenience for handling and consuming foods, rather than
on the existing recommendations. Any deviation from the suggested guidelines may likely
compromise the safety of ground beef and increase the risk of foodborne diseases. According
to the above, we aimed to evaluate the effect of frozen storage, thawing and cooking method of
beef patties on the survival of Salmonella and E. coli O157:H7, simulating common consumer-
style practices.

MATERIALS & METHODS

Portions (400 g) of ground beef were inoculated (~6.5 log CFU/g) with a five-strain composite
of Salmonella or a three-strain composite of Escherichia coli O157:H7 and stored at -22oC.
After 5 and 75 days of frozen storage, thawing took place as follows: (i) in refrigerator at 4oC
for 16 hours; (ii) at 20oC for 12 hours, simulating thawing on counter, or; (iii) in microwave for
22-24 minutes. Following thawing, 90 g beef patties were shaped and cooked by broiling or in
pan-grill up to two levels of internal temperature: 60oC, simulating undercooking or 71oC
(recommended cooking temperature). In addition, the survival of the two pathogens after direct
cooking of frozen patties was studied, simulating commercial cooking practices in catering
services and restaurants. The temperature of the samples during thawing or cooking was
measured with type-K thermocouples, which were fitted in the geometrical center of the
patties. Changes in the microbial populations that survived after cooking of patties were
monitored on TSA (total viable counts), CT-SMAC (E. coli O157:H7) or XLD (Salmonella).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1847
RESULTS & DISCUSSION
The populations of the pathogens exhibited slight reduction (0.5 – 0.7 log CFU/g) during
frozen storage for 5 or 75 days. This may occurred due to the mechanical injury of the cells
during freezing. The time-temperature profile during thawing of ground beef overnight (12
hours) at 20oC showed that the samples maintained in the “danger zone” (5-60oC) for over 7
hours. This extended exposure to ambient conditions resulted in an increase of 1 log CFU/g of
both pathogens. Cooking by broiling was more effective for eliminating 6.5 logs of both
microorganisms compared to cooking in pan-grill, especially when the target internal
temperature was 71oC. This may have occurred due to the slower heat transfer rate and the
longer exposure of the patties to heat during cooking in broiler compared to pan-grill (Figure
1). Undercooking (60oC) resulted only in 0.6 – 2.8 log CFU/g and 0.6 – 1.8 log CFU/g
decrease of Salmonella and E. coli O157:H7 populations, respectively, regardless of the
cooking method. Defrosting methods did not affect significantly (p<0.05) the survival of the
pathogens during subsequent thermal treatments. In contrast, frozen storage for 75 days
enhanced the survival of E. coli O157:H7, as the populations of the pathogen remained at high
levels, even when cooked by broiling at 71oC (1.7 – 3.1 log CFU/g).

(a) (b)

(d)
(c)

Figure 1 Changes of temperature in the geometrical center of beef patties during cooking in broiler (S- 60oC;
³ - 71oC) or pan-grill (Ƈ - 60oC; „ - 71oC), after thawing in refrigerator (4oC / 16 h; a), kitchen counter (20oC /
12 h; b), microwave (c) or without thawing (d)

CONCLUSION
The results may be used in the development and/or the revision of guidelines for the
appropriate practices, associated with defrosting and cooking of patties in households or
catering services.

1848
 European Food, Technology and Nutrition Declaration (EFTN Declaration)

Peter Raspora, Lidija Baša a

a
Chair of Biotechnology, Microbiology and Food Safety, Biotechnical Faculty, University of Ljubljana,
Jamnikarjeva 101, 1000 Ljubljana, Slovenia (peter.raspor@bf.uni-lj.si)

INTRODUCTION
History of human development illustrates clear image of impotence of food in our life. Food
has played important part in religion, is science, in technology, in medicine; not very pleasant
to hear but also in wars, in class distinction and relations between people. In everyday’s life we
have a “dialogue” with food (we have to eat, we have to buy food, we have to produce food,
ect). This “dialogue” has its benefits and its negative sides, that might be shown through
malnutrition on one side and overnutrition on other side, stable economy on one side and
unstable economy on other side, ect. There is always involved this balance catching issue.
From this point of view, the scientists and experts from field of food area initiated the EFTN
Declaration. EFTN Declaration’s aim is pointed at collaborative work among scientists and
experts who are dealing with challenging questions about food and nutrition in the Europe,
taking into account cooperative activities with the rest of the world. The inauguration event of
signing ceremony was held on 7 November, 2008 in City Hall in Ljubljana, Slovenia.

MATERIALS & METHODS

In this contribution we will avoid strict term materials & method since interted parties
contributed to the document development in written process. This was based on concept to
improve situation locally and globally. This was actually based on philosophy. The declaration
is not a paper of results where data would be gained by quantitative or qualitative method. It is
just a paper dealing with the broad area of food queries. EFTN Declaration`s main concept-
platform-philosophy is to bring together scientists, experts, those European professionals in
food science, technology and nutrition who are daily challenged by food dealing questions.
And what is the argument of this action? To found out the answer, its deepness and importance
please read the following quotation from the declaration:
“Food is necessary for human growth, development and the functions of the body. Good
nutrition demands a well-balanced diet that provides an adequate daily amount of all nutrient
classes and optimal intake of energy for the human body. Food must be safe, nutritious and
provided in a sustainable way that maintains consumer’s dignity and cultural identity (EFTN
Declaration, 2008).”

RESULTS & DISCUSSION

The first idea of the declaration raised in spring 2007. In the same year, 2007, in December
first call for Ambassadors was realised. To this action followed nomination of Ambassadors
(January 2008). First draft of declaration was prepared in June, 2008 and already in November
2008 the declaration was signed for the first time by Ambassadors from 41 different countries.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1849
Until now the declaration has been signed by EFTN Declaration`s Ambassadors from 43
different countries (Albania, Armenia, Austria, Belarus, Belgium, Bosnia and Herzegovina,
Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Georgia,
Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg,
Macedonia, Malta, Moldova, Montenegro, Netherlands, Norway, Poland, Portugal, Romania,
Russia, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey, Ukraine, United
Kingdom) and translated into 26 languages (Albanian, Armenian, Bosnian, Bulgarian,
Croatian, Dutch, English, Estonian, French, Georgian, German, Greek, Hungarian, Icelandic,
Italian, Lithuanian, Macedonian, Moldovan, Romanian, Russian, Serbian, Slovak, Slovenian,
Spanish, Turkish, Ukrainian). From the beginning of 2010 the EFTN Declaration is being
presented to the world through its website (http://eftndeclaration.aaeuropae.org).

CONCLUSION
For the conclusion let’s return to another quotation from the EFTN Declaration. This quotation
is about the cohesion of European food science, technology and nutrition.
“This declaration aims to initiate debate amongst European food scientists, food technologists
and nutrition professionals to establish mechanisms through which, hopefully, future
integration can be achieved. A second objective is to assist the harmonisation of the European
food industry and food quality and safety standards. All of these initiatives can help to promote
the expanding influence that the food science, technology and nutrition professions should
have on technological, scientific, political, environmental, social and cultural thought in
Europe. Of course, this must be done without harming nutrition and regional specificities of
food and diets in Europe since it is this diversity, which will generate future discoveries and
innovations (EFTN Declaration, 2008)”
There are different issues on food technology and nutrition. This is a permanent evolution and
novelties are coming in. This process will continue side by side with incremental development
in technology and habits. The consumer will stay in focus of food quality and food safety. But
it should be also in focus for food security. Unfortunately food reserves are going down and
prices are rising. In spite of this or better because of this issue the technology has many
possibilities; nutrition has many opportunities and general food sciences many challenges to
solve all this elements. This is real challenge to have better tomorrow with safe and wholesome
foods, but it will not help if we will not educate and train conscious consumer.

REFERENCES
[1] European Food Technology and Nutrition Declaration (EFTN Declaration).
http://eftndeclaration.aaeuropae.org, <10.01.2011>.
[2] Raspor, Peter. 2009. The European declaration on food, technology and nutrition: editorial. Acta
aliment. (Bp.), 1(38), 3-7.

1850
Optimization of shelf life distribution of frozen fish products based on modelling and TTI

monitoring

M. N. Giannogloua , M. Loukianou, K. Tsatsaragou, T. Tsironi and P.S. Taoukis

a
Laboratory of Food and Chemistry, School of Chemical Engineering, National Technical University of
Athens, 5 Iroon Polytechniou Str, 15780, Athens, Greece (giannoglou@chemeng.ntua.gr)

INTRODUCTION
Monitoring of storage temperature, the main shelf life determining post-processing parameter
of foods, constitutes an essential prerequisite for effective shelf life management. Application
of an optimized quality and safety assurance system for the distribution of frozen products
requires continuous monitoring and control of storage conditions. Time-Temperature
Integrators (TTI) are inexpensive, active “smart labels” that can show an easily measurable,
time-temperature dependent change that reflects the temperature history of a food product to
which it is attached.
The objectives of this study were the determination of the response function of two types of
UV activated photochemical and one type of enzymatic TTI labels, the development and
validation of kinetic models for the quality indices deterioration of the frozen fish products and
the evaluation of the applicability of a TTI based system to improve frozen fish product
quality.

MATERIALS & METHODS

Frozen shrimps and frozen sea-bream fillets were packed and stored at controlled isothermal
conditions from -15 to-5C. Samples were taken in appropriate time intervals for efficient
kinetic analysis of quality parameters (total viable count, pH, color, texture, lipid oxidation
(TBARS), total volatile basic nitrogen (TVB-N), trimethylamine nitrogen (TMA-N) and
sensory characteristics [1, 2, 3].
The OnVu-Logistics and the F4 photochemical prototype TTI labels(BASF, Germany) were
studied. Biserba GLP80 labelling unit (Biserba GmbH & Co. KG, Balingen, Germany), with
2” thin-layer thermal print head and TTF equipment was used for UV charging of the Logistics
for 0.2-0.8 s and 3-10 s for F4 TTI labels and subsequent laminating with a film (TTR 70QC)
which acts as an optical filter and protects TTI from light exposure and recharging.
M-type enzymatic TTIs (VITSAB,Sweden) with different enzyme concentrations ranging from
10 to 200 Units were studied.
For monitoring the colour change of a number of TTI tags, isothermally stored at constant
temperatures (from -15 to -5C), at appropriate time intervals, a colour measuring instrument
(X-Rite, Michigan, USA) was used displaying L, a and b values of the CIELab scale.

RESULTS & DISCUSSION

For monitoring the shelf lives of frozen shrimps and frozen sea-bream fillets, selected quality
indices (colour, texture, oxidative rancidity, TVBN and TMA analysis, aerobic plate count, pH
and sensory evaluation) were kinetically modelled and the temperature dependence of the
quality loss rates was modelled by the Arrhenius equation.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1851
For the selection of the appropriate photochemical and enzymatic TTI for monitoring the shelf
lives of the studied frozen fish products, TTI labels response was kinetically studied. For the
modelling of the response rate constant of the two photochemical TTIs as a function of degree
of initial activation and storage temperature, composite mathematical equations were
developed. A mathematical model which describes the effect of the concentration of the
substrate and the storage temperature on the response of the enzymatic TTIs was also
developed. Applying the developed models, the required charging levels for the photochemical
TTIs and the required substrate concentrations for the enzymatic TTI labels, to effectively
monitor the shelf life of the frozen fish products, were estimated.
Figure 1 shows the correlation between the shelf life of the frozen sea bream fillets and frozen
shrimps with the response times of two M-type TTI labels. M-17U and M-35U enzymatic TTI
labels response times are well correlated with the shelf lives of the frozen sea-bream fillets and
frozen shrimps, respectivelly.

50 0 M-17U CONCLUSION
M-35U
40 0 The selection of the appropriate TTI for
Shelf Lif e (da ys)

30 0 Sea-br eam monitoring the shelf lives of frozen

shelf life
20 0 Shrimps
shrimps and frozen sea breams fillets,
shelf life was based on developed mathematical
10 0
models. Applying the developed models,
0
the required charging levels for the
-20 -1 5 -1 0 -5 0
T (oC) photochemical TTIs and the required
substrate concentrations for the
Figure 1. Correlation of the response times of M-17U enzymatic TTI labels were estimated to
and M-35U TTI labels with the shelf lives of the frozen effectively monitor the shelf life of
sea-bream fillets and frozen shrimps frozen fish products.

Validation experiments measuring the response of the selected TTIs during a period of 12
months at various frozen temperatures are in progress.

REFERENCES
[1] Loovas E. A., Sensitive spectrophotometric method for lipid hydroperoxide determination, Journal of
the American Oil Chemists' Society 69 (1992), pp. 777–783
[2] Pivarnik L., Ellis P., Wang X. and Reilly T., Standardization of the Ammonia electrode method for
evaluating seafood quality by correlation to sensory analysis, Journal of Food Science 66 (7) (2001),
pp. 945–952.
[3] Tsironi T., Taoukis P., Modeling Microbial Spoilage and Quality of Gilthead Seabream Fillets:
Combined Effect of Osmotic Pretreatment,Modified Atmosphere Packaging, and Nisin on Shelf Life,
Journal of Food Science 75 (4) (2010), pp.243-251

Acknowledgement: This study was partly supported by the European Commission FP7 Collective
Research Project IQ-Freshlabel “Developing novel intelligent labels for chilled and frozen food products
and promoting the influence of smart labels application on waste reduction, food quality and safety in the
European supply chains” FP7-SME-2008-2-243423 (http://www.iq-freshlabel.eu).

1852
Modeling of Greek coffee aroma loss during storage at different temperatures and water

activities

E. Makri, D. Tsimogiannis, E.K. Dermesonluoglu, P.S. Taoukis

Laboratory of Food Chemistry and Technology, School of Chemical Engineering,

National Technical University of Athens, Athens, Greece (taoukis@chemeng.ntua.gr)

INTRODUCTION
Ground roasted coffee is a shelf-stable product. Because of the high temperature attained in the
roasting process, coffee is characterized by a very low water activity (aw) as well as the
presence of Maillard reaction products with antimicrobial properties [1]. However, during
storage, coffee may undergo important chemical and physical changes, responsible for coffee
“staling”, which affects the quality and acceptability [2]. The main causes of coffee staling are
attributable to losses of volatile compounds, in particular, of key sulfur-containing odorants
and oxidation reactions, the latter being responsible for off-flavor formation [3]. Temperature,
oxygen concentration and relative humidity/water activity are the major factors that affect the
shelf life of roasted coffee. The rate of coffee degradation reactions, may suddenly increase
after the packaging has been opened by the consumer, thus determining the so-called
“secondary shelf life”.
Coffee aroma, which involves more than 800 volatile compounds, is one of the most
contributory factors for the high acceptability of coffee by consumers. In fact, in the coffee
industry, sensory profiling is still the most widespread technique employed to evaluate the final
quality of both raw material and finished products. Several research groups have tried to
associate coffee staling with chemical changes in roasted coffee, obtaining the ratios between
certain pairs of volatile compounds, called aroma indices, which have been used as indicators
of coffee storage time [4].
The objective was to determine and characterize changes in the composition of the volatile
fraction of Greek coffee as a function of temperature and water activity during storage.

MATERIAL AND METHODS

Sample preparation
Greek coffee samples equilibrated at aw values, 0.15, 0.22, 0.33, 0.52, were stored at
temperatures T, 25, 35, 45°C. To maintain the water activity values, coffee samples were
placed in jars over saturated salt solutions (no salt -initial aw value-, CH3COOK, MgCl2,
Mg(NO3)2*6H20, Mg(NO3)2,ȱ respectively). Water activity was measured with a Rotronic A6
AM3t-AwVD (Basserdorf, Switzerland) aw-meter. To simulate home storage conditions, coffee
samples were not hermetically packaged.
Analysis of flavor compounds
The profile of volatile compounds was obtained directly from ground coffee samples using
purge & trap(OI Analytical 4660, USA)-gas chromatography-mass spectrometry methodology
(Agilent Technologies, USA, 5975C MSD System with Triple Axis Detector & 7890A GC
System). The storage temperature and water activity effect on volatile compounds was studied.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1853
RESULTS AND DISCUSSION
Volatile compounds identified and quantified throughout 150 days of storage at temperatures
studied were 7 aldehydes, 1 ester, 7 furans, 3 ketons, 1 pyrrole, 1 pyridine, 11 pyrazines and 1
alcohol. Among aldehydes detected, hexanal, n-decanal, n-heptanal and nonanal (malty flavor)
were the most abundant. The formation of hexanal (rancid flavor), due to the oxidation of
polyunsaturated fatty acids such as linoleic acid, seems to have a certain influence on the
staling of the coffee brew. It exhibited a stationary phase and then a rapid increase during
storage; however it should not be considered as a good marker. Among detected furans (burnt
sugar, caramel aromas), furfural that is the oxidative product of furfuryl alcohol showed an
exponential increase with storage that could be described mathematically (R2>0.90). In Fig. 1,
the change of furfural for coffee samples stored at T= 45°C and aw= 052 is representatively
shown. Within the group of ketones, special mention should be made of decrease of 2,3
pentanedione (buttery flavor). The most abundant pyrazine (roasty, earthy/musty flavors) was
2-methylpyrazine.
Increase of water activity (for aw values above 0.33) and increase of storage temperature (from
25 to 45°C) caused decrease of shelf life estimated based on selected aroma index for Greek
coffee. In Table 2 calculated shelf-life values for Greek coffee samples stored at different
temperature and water activity conditions were presented.
160 Table 1. Shelf-life values calculated based on aroma
retention for Greek coffee samples stored at T 25-45°C
140
and aw 0.15-0.52.
120
T, °C aw Shelf-life, days
100
Furfural

45 0.52 15
80
45 0.33 41
60 45 0.22 50
40 45 0.15 49
20 35 0.52 22
35 0.33 57
0
35 0.22 65
0 30 60 90 120 150
35 0.15 66
Storage Time (days)
25 0.52 88
Figure 1. Changes in furfural of coffee brews: 25 0.33 97
aw 0.52; T 25, 35, 45°C; t 150 days 25 0.22 98
25 0.15 104
CONCLUSIONS
The results of this study would give an idea about the possibility of the control of coffee
quality by measuring only a few volatile compounds. Furthermore, they can be used to define
tolerance on home storage conditions of Greek coffee.

REFERENCES
[1] Daglia, M., Cuzzoni, M.T., Dacarro, C. 1994. J. Agric. Food Chem., 42, 2270. [2] Nicoli, M.C.,
Savonitti, O. Espresso Coffee, 2nd ed.; Illy, A., Viani, R., Eds.; Elsevier Academic Press:San Diego, C.A.
2005. p.230. [3] Anese, M., Manzocco, L., Nicoli, M.C. 2006. J. Agric. Food Chem., 54, 5571. [4] Perez-
Martinez, M., Sopelana, P., Paz de Pena, M., Cid, C. 2008. J. Agric. Food Chem., 56(9), 3145

1854
 Combined effect of meat composition and heating parameters on the physicochemical

state of proteins

Promeyrat Aurélie, Le Louët Laure, Kondjoyan Alain, Astruc Thierry, Santé-Lhoutellier Véronique,
Gatellier Philippe, Daudin Jean Dominique

INRA, UR370 QuaPA, 63122 Saint Genès Champanelle, France (aurelie.promeyrat@clermont.inra.fr)

INTRODUCTION
During meat cooking, proteins undergo some oxidations and conformation changes which can
induce a loss in the nutritional value of products by a decreased bioavailability of essential
amino acids [1]. The present study aimed at determining the effect of heat treatments on the
physicochemical state of proteins (oxidation and thermal denaturation). To avoid confusing
effect due to the uncontrolled biological variability and to assess independently the specific
incidence of various compounds, we used two mimetic models: a basic model, composed of an
aqueous suspension of myofibrillar proteins, and a complex model, in which oxidants were
added in physiological concentrations. Heat treatments were applied at 4 temperatures (from
45°C to 90°C) during 5 times (from 5 to 120 min) and changes in proteins state were evaluated
by the measurement of carbonyl groups and protein surface hydrophobicity. Then, the results
were compared to those obtained on a pork meat model (M. Longissimus dorsi), cooked in the
same conditions.

MATERIALS & METHODS

1. Mimetic model making: The basic mimetic model was composed of myofibrillar proteins
in suspension in a phosphate buffer with a same pH and ionic strength than muscle (40 mM
and pH 6). Our objective was to determine kinetic laws of protein changes in pure fibre types
and to extrapolate these laws to mixed fibre types. Myofibrillar proteins were extracted from a
rabbit muscle (Psoas major, which is a 100% Į-white fibre type muscle), according to the
method of Pietrzak et al., 1997 [2] and delipidated by solvent (butanol/di-isopropyl ether)
which preserved the protein state.
Thermal treatments: Myofibrillar protein suspensions were heated at 45°C, 60°C, 75°C, and
90°C during 5, 10, 30, 60 and 120 min, in a dry bath.
Addition of oxidants: The basic mimetic model was complexified by addition, just before
heating, of a mixture of oxidants composed of hydrogen peroxide (2 mM), ascorbate (0.1 mM),
and ferrous iron. Iron concentrations of 0.05 mM, 0.2 mM and 0.6 mM, were tested to screen
different lean meat tissues. These oxidants produce hydroxyl radicals (OH°), by the Fenton
reaction, which are very reactive towards proteins.
2. Pork meat model: To avoid biological variability of raw meat the experiments were carried
out on samples taken from only one pig muscle Longissimus dorsi (fibre type: 11% ȕ-red, 16%
Į-red, 73% Į-white). Thin slices of meat (1.5 mm) were enclosed in plastic bags and heated in
a water bath using the same conditions than with the mimetic models. Myofibrillar proteins
were prepared from the heated samples using the same protocols as described above.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1855
3. Characterisation of protein physicochemical changes: Protein oxidation was assessed by
the measurement of carbonyl groups [3] and protein thermal denaturation was evaluated by the
protein surface hydrophobicity [4].

RESULTS & DISCUSSION

Protein oxidation: In proteins, carbonyl groups are formed by oxidation of basic amino acids,
which are essential amino acids for humans. Moreover, carbonyls groups can react with free
amino groups of non oxidized amino acids of proteins to form amide bonds which are
precursors of protein aggregation. The results obtained with the two mimetic models showed
that carbonyls can not be produced under the thermal process alone; oxidants are required for
their formation. A synergic effect of the chemical and thermal treatments was noticed on
protein oxidation. Carbonyl production was considerably higher in the complex mimetic model
than in meat and this can be explained by the muscle antioxidant protection which was not
taken into account in this preliminary version of the mimetic model.
Protein surface hydrophobicity: Thermal treatment can impair the structure of meat proteins
by breaking of hydrogen and electrostatic bonds. As a consequence of this thermal
denaturation, an exposure to the protein surface of hydrophobic amino acids can occur and
favour a tendency of proteins to form aggregates. The mimetic models results showed that
protein surface hydrophobicity was mainly under the dependence of thermal denaturation and
that oxidants poorly affected its change. In contrast to protein oxidation, hydrophobicity values
were higher in meat than in the mimetic model. In general, at highest temperatures, thermal
denaturation is a rapid process, the maximum value of hydrophobicity was reach from 5
minutes and no further increase was observed with increasing heating time.

CONCLUSION
The main advantage of using the mimetic models was to assess the relative contributions of the
chemical and thermal effects on protein changes. Our ultimate goal is to predict through a
mathematical tool the effects of heat treatments on the nutritional value of meat products.
Thus, to build up this tool the kinetics of protein oxidation and denaturation will be modelled
and combined to heat-mass transfer models to consider realistic cooking conditions in large
pieces of meat.
Acknowledgement: The research leading to these results has received funding from the
European Community's Seventh Framework Programme (FP7/ 2007-2013) under the grant
agreement n°FP7-222 654-DREAM.

REFERENCES
[1] Santé-Lhoutellier V., Aubry L. & Gatellier, P. 2007. Effect of oxidation on in-vitro digestibility of
skeletal muscle myofibrillar proteins. Journal of Agricultural and Food Chemistry, 55, 5343-5348.
[2] Pietrzak M., Greaser M.L. & Sosnicki A.A. 1997. Effect of a rapid rigor mortis processes on protein
functionality in Pectoralis major muscle of domestic turkeys. Journal of Animal Science, 75, 2106-
2116.
[3] Oliver C.N., Alin B.W., Moerman E.J., Goldstein S. & Stadtman E.R. 1987. Age-related changes in
oxidized proteins. Journal of Biological Chemistry, 262, 5488-5491.
[4] Chelh I., Gatellier P. & Santé-Lhoutellier V. 2006. Technical note: A simplified procedure for
myofibril hydrophobicity determination. Meat Science, 74, 681-684.

1856
 Biogenic amine levels in dry fermented sausages produced and sold in Greece

E.J.Papavergoua

a
Laboratory of Food Technology, Department of Food Hygiene & Food Technology of Animal Origin,
Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Greece (ekpap@vet.auth.gr)

INTRODUCTION
Biogenic amines (BAs) are formed in foods by enzymatic decarboxylation of free amino acids
produced during fermentation and/or aging of foods. Fermented sausages may contain high
levels of BAs due to the high protein content of raw materials and also their production
technology which includes a microbial fermentation period followed by a long ripening period
during which extended proteolysis occurs [1]. Since BAs can cause various food-born diseases
to consumers and also, their formation in fermented foods may indicate the presence of
undesired contaminating microbial flora, there is an increased need for monitoring their
presence in fermented sausages. The present study was carried out in order to determine the
levels of BAs in dry fermented sausages produced and consumed in Greece. The amines
measured include putrescine (PU), cadaverine (CA), tyramine (TY), histamine (HI), tryptamine
(TR), ȕ-phenyl-ethylamine (PHE), spermidine (SD) and spermine (SP).

MATERIALS & METHODS

Forty samples of various types of dry fermented sausages made in Greece by different
manufacturers were bought from retail markets of Thessaloniki during the summer of 2010.
BAs were extracted from the samples with 0.6 M HClO4 and subsequently measured by HPLC
using fluorescence detection after post column derivatisation with o-phthaldialdehyde [2].

RESULTS & DISCUSSION

The formation of BAs during the production of dry fermented sausages is mainly attributed to
the decarboxylating activity on free amino acids of certain fermenting or contaminating
microbial groups present in them such as several Lactobacillus and Enterococcus strains
(formation of TY, PHE, PU), Pseudomonas spp, (formation of PU), genera of the family of
Enterobacteriaceae, (formation of PU, CA and HI) and of Micrococcaceae (formation of HI,
PHE) [1]. Various fermentation and ripening parameters such as high availability of free amino
acids due to extended proteolysis, prolonged ripening period, pH decrease, increased
fermentation and storage temperatures and potentially high microbial load of raw materials also
favour the accumulation of BAs in fermented sausages [1].
The levels of BAs measured in the examined samples are summarized in Table 1. A wide
variation of BA concentrations was observed among the samples. In most of them, the BAs
present in the highest concentrations were PU or TY. Similar TY and PU levels to those found
in the present study were reported in traditional fermented sausages manufactured in other
countries [1]. However, selection of bacterial starter cultures with limited tyrosine and
ornithine decarboxylating activity might reduce TY and PU levels in fermented sausages [3].
CA concentrations were lower and exceeded 100 ȝg/g in only 17.5% of the examined samples.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1857
High CA contents were usually accompanied by high PU levels in the same sample indicating
either poor hygienic conditions and freshness of raw materials, or adoption of incorrect
production procedures by the manufacturer [1]. HI levels were low in most of the examined
samples. However, a notable proportion of the samples (37%) far exceeded the toxicity limit of
100 mg/kg legally set for HI in some fish species. High initial microbial load of raw materials,
inadequate decrease of pH at the beginning of the ripening process, or extended ripening time
could be the reasons for considerable accumulation of HI in some of the fermented sausages.

Table 1. Biogenic amine contents (mg/kg) in fermented sausages produced and consumed in Greece
Amines Range Median Mean ± S.D.
TY 3.66 – 381.43 175.19 164.95 ± 98.08
PU 0 – 491.74 176.62 187.86 ± 144.94
CA 0 – 1014.08 15.31 94.19 ± 213.62
HI 0 – 375.75 36.28 89.07 ± 101.55
TR 0 – 60.53 6.81 14.27 ± 17.61
PHE 0 – 56.40 2.81 6.71 ± 11.24
SD 1.45 – 19.53 5.72 6.71 ± 3.98
SP 13.37 – 60.11 35.92 36.63 ± 10.30
TY+HI+PHE 5.56-619.48 264.7 261.29 ± 176.37

TY, HI and PHE show vasoactive and psychoactive properties and foods containing them in
high levels may constitute a potential risk for certain sensitive groups of consumers exhibiting
deficient biogenic amine metabolising ability [4]. The sum of these three amines exceeded in
63% of the examined samples the proposed for fermented sausages limit of 200 mg/kg [1] for
good manufacturing practices and safe consumption. The presence of high contents of PU and
CA found in many of the examined samples might further potentiate the toxicological impact
of TY, HI and PHE in them [4].

CONCLUSION
It is concluded that an improvement in production technology and raw material hygienic
quality used for manufacturing various types of dry fermented sausages in Greece may be
necessary in order to ensure the safe consumption of this type of meat products with respect to
their BA content.

REFERENCES
[1] Suzzi G. & Gardini F. 2003. Biogenic amines in dry fermented sausages: a review. International
Journal of Food Microbiology, 88(1), 41-54
[2] Hernández-Hover T., Izquierdo-Pulido M., Veciana-Nogués M.T. & Vidal-Carou M.C. 1996. Ion-
Pair High-Performance Liquid Chromatographic Determination of Biogenic Amines in Meat and
Meat Products. Journal of Agricultural and Food Chemistry, 44(9), 2710-2715.
[3] Komprda T., Sládková P. & Dohnal V. 2009. Biogenic amine content in dry fermented sausages as
influenced by a producer, spice mix, starter culture, sausage diameter and time of ripening. Meat
Science, 83(3), 534-542.
[4] Shalaby A.R. 1996. Significance of biogenic amines to food safety and human health. Food Reasearch
International, 29(7), 675-690.

1858
 Spore Inactivation by Ultraviolet Irradiation Combining with Different Pre-heating

Treatment

Daisuke Hamanakaa, Hironori Yamadab, Takashi Kadoyanagib , Vipavee Tryvittayasilb,

Fumihiko Tanakaa, Toshitaka Uchinoa

a
Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku,
Fukuoka 8128581, Japan (hamanaka@bpes.kyushu-u.ac.jp)
b
Graduate School of Biores. Bioenviron. Sci., Kyushu University, 6-10-1
Hakozaki, Higashi-ku, Fukuoka 8128581, Japan

INTRODUCTION
The disinfecting treatment by ultraviolet ray (UV) irradiation has been widely applied in
various industrial fields [1]. In relation to the preservation of fresh agricultural produce, several
researchers investigated the application of UV irradiation to surface decontamination. The
inactivating efficiency of UV irradiation was enhanced by the combination of other
decontaminating techniques. Some researchers tried to extend the shelf life of fresh fruits by
the combining treatment of UV irradiation and thermal technologies [2-3]. However, no
information on the effect of the treatment temperature controlled by infrared radiation heating
(IRH) on the inactivating efficiency of UV irradiation was reported. In the present study, we
investigated the influence of IRH temperature in addition to UV irradiation for microbial spore
inactivation on agar media. We also compared with the killing efficiency of UV irradiation
under the same temperature conditions prepared by conductive heating (CH) using an electric
hot plate.

MATERIALS & METHODS

The sample microbial spores (Penicillium isolated from peach fruit and Bacillus subtilis
NBRC3134) inoculated onto the surface of agar media (potato dextrose agar, nutrient agar)
plate were heate d to the required temperatures (20, 30, 40, and 50 °C) by either CH or IRH,
and then exposed to UV light for required periods by laboratory assembled devices (Figure 1).
The surface temperature of agar media was monitored by infrared thermography camera.
Surface temperature profile of IRH was adjusted by the current setting of IR heater using an

㻔㻭㻕 㻔㻮㻕

a a
150mm
b 150mm
c c

d
e
Figure 1. Schematic diagrams of experimental devices. The distribution of irradiation
intensity of UV at the surface of sample plate was ensured to be uniform at both devices.
a: ultraviolet lamp, b: infrared radiation heater, c: ȭ90mm sample plate (potato dextrose
agar/nutrient agar), d: table, e: electric hot plate

11th International Congress on Engineering and Food - Athens, Greece, 2011 1859
electric transformer in order to give same profile of CH treatment.

RESULTS & DISCUSSION

The survival curves of spores of Penicillium and B. subtilis did show convex upward and
downward curves, respectively. Generally, the inactivation efficiency of UV irradiation with
CH (UV-CH) was greater than that with IRH (UV-IRH) for Penicillium spores. The greatest
inactivat ion, 2.3 logs reduction, was obtained by 60 sec of UV-CH treatment at 20 °C. In
contrast, the inactivation efficiency of UV-IRH was greater than that of UV-CH for B. subtilis
spores. A 4.1 log reduction of B. subtilis spores was obtained by 60 sec of UV-IRH treatment
at 50 °C. The Duv-values (the times required for 90% reduction) of Penicillium spores appeared
to be greater with the increase of temperature in both treatments of UV-IRH and UV-CH. The
greater Duv -values of B. subtilis spores were obtained at a mild temperature range (around
30 °C), especially with the treatment of UV-CH (Figure 2).
40
㻰㼑㼏㼕㼙 㼍㼘㻌㼞㼑㼐㼡㼏㼠㼕㼛㼚㻌㼠㼕㼙 㼑㻌㻔㼟㼑㼏㻕

30

20

10

0
10 20 30 40 50 60
㼀㼑㼙 㼜㼑㼞㼍㼠㼡㼞㼑㻌㻔䉝㻕
Figure 2. Relationship between decimal reduction time (Duv-value) of Bacillus
subtilis spores and temperature controlled by conductive heating (‫ )ۑ‬and infrared
radiation heating (‫)ڦ‬. Duv-values were calculated by the linear regression portion of
the survival curves.

CONCLUSION
It is suggested that these obtained differences in the inactivation characteristics of UV
irradiation could be caused by the differences in UV absorption characteristics of microbial
spores treated by different thermal processes.

REFERENCES
[1] Bintsis, T., Litopoulou-Tzanetaki, E. & Robinson, R.K. 2000. Existing and Potential Applications of
Ultraviolet Light in The Food Industry – a critical review. Journal of the Science of Food and
Agriculture, 80, 637-645.
[2] Marquenie, D., Michiels, C.W., Geeraerd, A.H., Schenk, A., Soontjens, C., Van Impe, J.F. & Nicolai,
B.M. 2002. Using Survival Analysis to Investigate The Effect of UV-C and Heat Treatment on
Storage Rot of Strawberry and Sweet Cherry. International Journal of Food Microbiology, 73, 187-
196.
[3] Pan, J., Vicente, A.R., Martinez, G.A., Chaves, A.R. & Civello, P.M. 2004. Combined Use of UV-C
Irradiation and Heat Treatment to Improve Postharvest Life of Strawberry Fruit. Journal of the
Science of Food and Agriculture, 84, 1831-1838.

1860
 Aroma profile of different salted dried codfishes

Marta Costa Silvaa, Luís R. Silvab*, Paula Guedes-de-Pinhoc, Paula Andradeb, Patrícia Valentãob, Rui
Costaa*
a
CERNAS/ Escola Superior Agrária, Instituto Politécnico de Coimbra, Coimbra, Portugal
(ruicosta@esac.pt)
b
REQUIMTE/ Dept of Pharmacognosy, Faculty of Pharmacy, Porto University, Porto, Portugal
(ext_lsilva@ff.up.pt)
c
REQUIMTE/ Department of Toxicology, Faculty of Pharmacy, Porto University, Porto, Portugal

INTRODUCTION
Salted dried codfish is a highly consumed product in Mediterranean and South American
countries. Several procedures are involved in the traditional manufacturing of this product that
brings up organoleptic differences. Aroma is the first characteristic that influences consumer
perception of codfish quality at the selling point and latter at the consumption stage.
The aim of the present study was to evaluate the differences in volatile profile of salted dried
codfishes species commercialized in Portugal, caught in different fishing zones, and cured by
different processes and for distinct periods. For these purposes HS-SPME/GC IT-MS analysis
was performed, data obtained was treated by using a principal component analysis (PCA).

MATERIALS & METHODS

Volatile profiles of three species (G. morhua, G. macrocephalus and T. chalcogramma) caught
in distinct origins along with two curing processes (Traditional and Yellow curing) and
different processing times, were characterized by HS-SPME/GC IT-MS. The codfish samples
were supplied by Lugrade (Coimbra, Portugal).
Each sample was magnetically stirred (200 rpm) at 40 °C, for 10 min. The fibre coated with
divinylbenzene/polydimethylsiloxane (DVB/PDMS - 65 ȝm) was exposed to the headspace for
30 min, with agitation (200 rpm) at 40 ºC. Afterwards the fibre was inserted into the injection
port of the GC system for thermal desorption, for 1 min. The fibre was then conditioned in
another GC injection port for 10 min at 250 °C.
Principal component analysis (PCA) was carried out using XLSTAT 2010.3.01 software.

RESULTS & DISCUSSION

The analysis by HS SPME/GC-IT-MS of salted cured codfish samples allowed the
characterization of fifty-five volatile compounds, which were distributed by distinct chemical
classes: two amines, two hydrocarbons, one ester, one chlorinated compound, ten aldehydes,
eight aromatic hydrocarbons, sixteen alcohols, three acids, one sulphur compound, two ethers,
three ketones, two terpenes and four other compounds. The major class of identified
compounds in the analyzed samples was that of alcohols (ca. 29.1 % of total identified
compounds), followed by aldehydes (ca. 18.2%) and aromatic hydrocarbons (14.5%). Esters,
chlorinated and sulphur compounds represented the minor components (<1.8% each).
Many of the compounds here identified were already reported in codfish aroma profiles [1, 2].
The highest diversity of compounds was found in yellow cured codfish sample (MOIY) and
traditional cured codfish sample from Canada zone (MOC), with fifty one compounds being
determined. Yellow cured codfish (MOIY) showed the highest amount of volatiles, being

11th International Congress on Engineering and Food - Athens, Greece, 2011 1861
richer in compounds from all chemical classes excepting amines, terpenes, sulphur compounds
and acids. Amongst all, salted dried codfish from Russian/Norway fishing zone (MOR)
exhibited the poorer volatiles profile, with thirty four compounds being characterized.
To assess the variation of volatile composition of the analyzed codfish samples, PCA was
performed (Fig. 1).

Variables (axes F1 and F2: 73,80 %) Observations (axes F1 and F2: 73,80 %)
1
4
Amines
0,75
Ethers 3
MOC
0,5
Sulphur
compound 2 MAP7

0,25 Ketones
Acids
F2 (23,42 %)

F2 (23,42 %)
Aldehydes 1
Alcohols MAP
0
Others
Aromatics 0
MOR
Ͳ0,25 hydrocarbons MOI
Chlorinated
compound Ͳ1
MOIY
Ͳ0,5 Esters
Hydrocarbons
Ͳ2
Ͳ0,75 Terpenes

TCP
Ͳ3
Ͳ1
Ͳ3 Ͳ2 Ͳ1 0 1 2 3 4 5 6
Ͳ1 Ͳ0,75 Ͳ0,5 Ͳ0,25 0 0,25 0,5 0,75 1

F1 (50,38 %) F1 (50,38 %)

Figure 1 - PCA of the volatile profiles of salted dried codfish samples: projection of volatile compounds
into the plane composed by the principal axes F1 and F2 (73.80%).

CONCLUSIONS
Thirty compounds were fully characterized and 25 were tentatively identified, from which 26
are reported for the first time in dried salted codfish aroma, contributing to a greater knowledge
of their volatile composition. Differences among fish species and fishing zones were not
perceivable, with the exception of T. chalcogramma (TCP) that presented more terpenes and
less aldehydes than the other samples. Microbial activity, know to be much higher in the
yellow curing, appears to have a high contribution to the overall production of volatiles.

REFERENCES
[1] Olafsdottir G., Jonsdottir R., Lauzon H.L., Luten J., & Kristbergsson K. 2005. Characterization of
volatile compounds in chilled codfish (Gadus morhua) fillets by gas chromatography and detection of
quality indicators by an electronic nose. Journal of Agricultural and Food Chemistry 53,10140-10147.
[2] Guillén M.D., Errecalde M.C., Salmerón J. & Casas C. 2006. Headspace volatile components of
smoked swordfish (Xiphias gladius) and codfish (Gadus morhua) detected by means of solid phase
microextraction and gas chromatography – mass spectrometry. Food Chemistry 94, 151-156.

1862
 Influences of pH and temperature on infrared spectroscopic features of brewed coffee

Atsushi Hashimoto, Yoshiko Sugimoto, Ken-ichiro Suehara, Takaharu Kameoka

Department of Sustainable Resource Sciences, Graduate School of Bioresources, Mie University, Tsu, Japan
(hasimoto@bio.mie-u.ac.jp)

INTRODUCTION
A huge amount of brewed coffee is manufactured, and very long time has passed since the start of the
manufacturing. However, the coffee process is empirically controlled based on the information provided
by the cup tasters and the manufacturing experts, and the process control could be generally due to the
expertise accumulated in each coffee manufacturing company. Quality of brewed coffee highly depends
on the chemical components and their interactions. The states of the chemical components and their
interactions are significantly influenced by the temperature and pH.
The application of spectroscopy, especially in mid-infrared (MIR) region, to the above measurement is
then desirable as a high potential implement. The objective of this study is to develop a simple, rapid,
accurate and non-chemical method evaluating the brewed coffee by MIR spectroscopy. We studied the
influences of the pH and temperature on the MIR spectral characteristics of the brewed coffee by
considering hot and iced brewed coffee and by spectroscopically discussing the spectral features of the
brewed coffee model composed from the main component.

MATERIALS & METHODS

Aqueous solution of the commercial instant coffee was prepared as brewed coffee. Additionally,
chlorogenic acid, citric acid, quinic acid, trigonelline, caffeine and arabinogalactan were selected as the
main components in brewed coffee, and their aqueous solution and the mixture (brewed coffee mode;
Table 1) were prepared.
Table 1. Concentrations of reagents in brewed coffee model.
Reagents Concentration [mg/100 ml]
chlorogenic acid 37
citric acid 20
ionic dissociative components
quinic acid 46
trigonelline 10
caffeine 60
ionic undissociative component
arabinogalactan 300

The infrared spectroscopic measurement system [1, 2] was used for the spectral measurements, which is
composed from the FT-IR system (Magna-IR 750; Nicolet Instrument Corp.), the cylindrical sample
container, a water bath and a data logger. The bottom part of the cylindrical sample container was the
ATR accessory (DuraSampleIR; SensIR Technologies) equipped the FT-IR system, and the cylinder
made of stainless steel adheres to the top of the ATR accessory. The ATR spectra were measured at 283,
298 or 333 K.

RESULTS & DISCUSSION

The influences of the pH and temperature on the second derivative spectral characteristics of brewed
coffee are displayed in Figure 1. The absorption peak shifts and the absorption intensity changes due to

11th International Congress on Engineering and Food - Athens, Greece, 2011 1863
the pH and temperature changes are observed. The ionic equilibrium conditions of the ionic dissociative
materials such as chlorogenic and quinic acids as the main components in the brewed coffee could be
influenced by the pH value, and could be also changed due to the temperature at the same pH value.
In order to study spectral behavior of brewed coffee, the spectra theoretically calculated based on the
spectral additivity and the ionic dissociation equilibrium conditions were compared with measured ones
under several pH and temperature conditions [3, 4]. Figure 2 shows influences of the simultaneous
changes of the temperature and pH on the experimental and calculated infrared absorption spectra of the
brewed coffee model. The calculated spectra represented the infrared absorption peaks characterizing the
brewed coffee model as well as the measured ones. Additionally, the peak shifts due to the temperature
and pH changes of the brewed coffee model of the calculated spectra consistently agree with the
experimental ones. These results suggest that the spectral analyzing method of the brewed coffee model
developed in this study could be acceptable to spectroscopically understand the quality in consideration of
the temperature and pH.

Figure 1. Influences of pH and temperature Figure 2. Influence of pH and temperature

on infrared absorption spectra of brewed coffee. on actual and calculated second derivative
and calculated second derivative spectra
of brewed coffee model.

CONCLUSION
This study is an important step in the developments of the quality and taste of brewed coffee based on the
MIR fingerprint spectra, which include the information of the chemical components and the interactions
in brewed coffee, and of the non-intensive on-line monitoring of the brewed coffee production process.

REFERENCES
[1] Pan, T., Hashimoto, A., Kanou, M., Nakanishi, K. & Kameoka, T. 2003. Development of a Quantification System
of Ionic Dissociative Materials in the Glycolytic Pathway Using an FT-IR/ATR Method. Bioprocess and
Biosystems Engineering, 26(2), 133-139.
[2] Hashimoto, A., Pan, T., Kanou, M., Nakanishi, K. & Kameoka, K. 2005. Mid-Infrared Spectroscopic Monitoring of
Enzyme Reaction Associating with Ionic Dissociative Metabolites. In: Pons, M.-N. & van Impe, J.F.M. (Eds.).
Computer Applications in Biotechnology 2004. Elsevier Science, Oxford. pp.375-380.
[3] Nakanishi, K., Hashimoto, A., Kanou, M., Pan, T. & Kameoka, T. 2003. Mid-Infrared Spectroscopic Measurement
of Ionic Dissociative Materials in Metabolic Pathway. Applied Spectroscopy, 57(12), 1510-1516.
[4] Hashimoto, A., Sugimoto, Y., Suehara, K., Kanou, M. & Kameoka, T. 2009. Infrared Spectroscopic Analysis on
Wine Components Using Ionic Dissociative Information. 5th International Conference on Advanced Vibrational
Spectroscopy, Melbourne, Australia, 12-17 July 2009. Book of Abstract, pp. 117-118.

1864
 Comparison of wild and farmed sea bass (Dicentrarchus labrax L) lipid quality

Lenas Dimitriosa, Chatziantoniou Soumelab, Nathanailides Cosmasa, Triantafillou Dimitriosb

a
Dept Aquaculture & Fisheries, TEI of Epirus, Igoumenitsa, Greece, (Lenasds@teiep.gr)
b
Alexander Technological Institute of Thessaloniki, Dept Nutrition & Dietetics, Thessaloniki, Greece
(tridim@nutr.teithe.g)r

INTRODUCTION
The nutritional value of fish is attributed to the amino-acids, trace elements, vitamins, but most
importantly, to their fatty acids (FAs). Recently, a reduction in the utilisation of fish oil in the
manufacturing of aquaculture feeds is reflected in the FAs of farmed fish. The aim of the
present work was to compare the fatty acids profile of wild and farmed sea bass flesh.

MATERIALS & METHODS

Wild and farmed sea bass specimens were obtained from a fisherman and a fish farm in NW
Greece. The feed (manufacturer label) chemical analysis was: 45% proteins, 20% lipids, 14.5%
carbohydrates, 9% ash, 1.5% fibres, 10% moisture, vitamins and trace elements. The raw
materials were fish meal, fish oil, wheat flour, corn gluten, vitamins and minerals (no
indication of %). Fillets were obtained and lipid was extracted using chloroform and methanol.
The fatty acids were methyl-esterified in 12% (BF3-MeOH). Methyl-esters were obtained with
normal hexane for gas chromatography.

RESULTS & DISCUSSION

The dominance of 18:2n-6, linoleic acid (LA) the increased 18:3 n-3, Linolenic acid (LNA)
and the increased n-6/n-3 ratio in the flesh in farmed fish appears to be a result of the inclusion
of increased amounts of plant oils as an alternative raw material for fish feeds [1]. As a result
farmed fish exhibited lower nutritional value in terms of FAs, especially in terms of high LA&
LNA content and low n-3/n-6 ratio, which is important for human’s health. LA and LNA in
fish feeds cannot be transformed easily to n-6 and n-3 FAs in the flesh of marine fish, contrary
to fresh water fish. This is probably due to lack of necessary enzymes (D-5-6-Desaturase)
which elongate the carbonic chain of fatty acids [5]. This disability of marine fish leads to LA
and LNA inclusion in flesh lipids, which finally end to the human body through consumption.
The main reason for the use of alternative sources of raw materials in fish feeds is the high cost
of fish meal and fish oil due to increasing demand and limited production from wild fish
stocks. This trend tends to increase as a convenient tool to reduce production cost in an
intensely competitive market and have been reflected in previous published workss [1,4] and in
FAO reports for decreased imports and consumption of fish meal and fish oil, which during the
last decade, decreased by 18% in the European Union, despite the increase of fish farming
production. This fact indicates that the alteration of fish feeds composition have been
established in the at least during last decade. Nutritional experiments in fish showed that the
addition of 60% soybean oil in fish feed resulted in a 428-647% increased levels of LA in the
flesh of fish compared to fish oil based feed. In the same manner, 60% linseed oil resulted in
increased levels of LA in the flesh of fish compared to controls [3]. Increased LA consumption

11th International Congress on Engineering and Food - Athens, Greece, 2011 1865
may cause 100 times greater risk of homicide mortality. Increased LA consumption may
contribute to depression and increased cardiac mortality [2].

Table 1. Fatty acid profile (% ±SD) of wild and farmed sea bass lipids
Fatty acid Wild Sea bass Farmed Sea bass
C14:0 2.34±0.11 2.75±0.14
C15:0 0.64±0.03 0.33±0.04
C16:0 17.69±0.40 13.81±0.63
C18:0 5.40±0.62 3.72±0.15
C16:1 n-7 (9C) 5.72±0.28 3.97±0.19
C18:1 n-9 (9C) 20.01±1.94 19.56±0.49
C18:1 n-7 (11C) 4.55±0.47 3.15±0.20
C20:1 n-9 (11C) 0.00±0.00 0.41±0.06
C22:1 n-9 (13C) 7.55±0.36 6.71±0.14
C18:2 n-6 4.42±0.29 18.02±o.12
C18:3 n-3 1.53±0.03 2.08±0.10
C18:4 n-3 0.62±0.02 0.69±0.06
C20:4 n-6 3.63±0.23 0.68±0.04
C20:5 n-3 0.00±0.00 2.15±0.08
C22:4 n-6 0.00±0.00 0.12±0.02
C22:5 n-3 2.96±0.13 1.81±0.06
C22:6 n-3 15.41±0.63 8.60±0.17
Total n-3 Fatty acids 20.52±0.54 15.33±0.44
Total n-6 Fatty acids 8.05±0.51 18.81±0.18
Ratio n-3/n-6 2.55±0.10 0.81±0.02

CONCLUSION
The increased LA, LNA and n-6 of farmed fish can be attributed to the increased amounts of
plant oils in fish feeds manufacturing. This feed driven change of farmed fish works against the
public health and those who consume it for the cardiovascular benefits of a high n-3/n6 diet.

REFERENCES
[1] Dubois V., Breton S., Linder M., Fanni J., & Parmentier M. 2007. Fatty acid profiles vegetable oils
with regard to their nutritional potential. European Journal of Lipid Science & Technology, 109, 710-732.
[2] Hibbeln J.R., Nieminen L.R.G., Blasbalg T.L., Riggs J.A., & Lands W.E.M. 2006. Healthy intakes of
n-3 and n-6 fatty acids: estimations considering worldwide diversity. American Journal of Clinical
Nutrition, 83, 1483S-1493S. [3] Izquierdo M.S., Montero D., Robaina L., Caballero M.J., Rosenlund G.,
& Gines R. 2005. Alterations in fillet fatty acid profile and flesh quality in gilthead sea bream feed
vegetable oils for a long term period. Recovery of fatty acid profiles by fish oil feeding. Aquaculture, 250,
431-444. [4] Phichova J. & Morkore T. 2007. Alternate oils in fish feeds. European Journal of Lipids
Technology, 109, 256-263. [5] Tocher D.R. (2003). Metabolism and functions of lipids and fatty acids in
teleost fish. Reviews in Fisheries Science, 11(2), 107-184.

1866
 Coupling between heat and mass transfer and stoechio-kinetic models to bring insight

into Maillard reaction kinetics during baking of sponge-cake products

Caroline Pénicaud b,a, Bertrand Broyarta,b, Daniel Goujotb,a , Mathilde Courela,b, Xuan-Mi Meyerc,
Catherine Bonazzib,a

a
AgroParisTech, UMR 1145 Ingenierie Procedes Aliments, Massy, France
(bertrand.broyart@groparistech.fr)
b
INRA, UMR 1145 Ingenierie Procedes Aliments, Massy, France (catherine.bonazzi@groparistech.fr)
c
Université de Toulouse, Laboratoire de Génie Chimique CNRS/INPT/UPS, Toulouse, France
(XuanMi.Meyer@ensiacet.fr)

INTRODUCTION
The present work is part of the REACTIAL project "Prediction and control of the appearance or
disappearance of reactional markers during food process and conservation" (ANR-06-PNRA-023)
supported by the French National Research Agency. Very few studies have taken into account both
effect of heat and mass transfer phenomena and composition of the product (in terms of chemical
species involved) upon nature and extent of Maillard reaction within solid food products undergoing
“realistic” heat treatments. One of the major issues of this work is hence to extract an “apparent
reaction scheme” from the complex Maillard reaction scheme. This apparent reaction scheme must
be: (1) able to give reliable representation of the real complex reaction scheme of Maillard reaction
(2) coherent with analytical capabilities by isolating so-called chemical markers representative of
the extent of Maillard reaction. From this apparent reaction scheme, stoechio-kinetic models will be
developed. These equations will appear as sink- and source-terms in classical heat and mass transfer
continuity equations in order to predict local kinetics of product temperature rise and moisture loss
as well as kinetics of variation of concentrations of chemical species appearing in the reaction
scheme. The aim of this work is to give an overview of the results concerning the prediction of
extent of Maillard reaction during sponge cake baking coupled with the prediction of product
temperature rise and moisture loss kinetics during baking. Main hypotheses of both physical and
chemical models will be given as well as constitutive equations and preliminary results of
validation.

MATERIALS & METHODS

Baking experiments of sponge-cake and associated chemical analyses (including vapour products
present in the baking atmosphere) were realised using a specifically instrumented electric
convective oven presented in [1]. Oven atmosphere was continuously extracted from the oven cavity
and analysed using dynamic headspace-solid phase microextraction (HS-SPME) and GC-MS
system adapted to the present oven from a previous study [2]. Seven Maillard reaction markers were
followed during baking among which non-volatile markers measured in the product (glucose, amino
groups and 5-hydroxymethylfurfural HMF) and volatile markers measured in the baking atmosphere
(furfural, 2,3-dihydroxy-6-methyl-pyranone or DDMP, acetic and formic acids and HMF).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1867
RESULTS & DISCUSSION
Concerning the modelling of heat and mass transfer phenomena, the cereal product is considered as
a biphasic medium constituted of a continuous “pseudo-liquid” phase (L) containing liquid water,
dissolved chemical markers of Maillard reaction and dry matter, locally in equilibrium with a
gaseous phase (G) composed of vapour and volatile chemical markers. The heat and mass transport
phenomena in the product are assumed unidirectional. The upper surface of the product is heated by
convection and radiation and its lower surface partly by contact with a perforated baking tray and
partly by convection and radiation. The value of product porosity is assumed uniform within the
product and constant during baking. The product is assumed non-shrinking during baking. The
porosity of the product is considered as an open porosity allowing gas diffusion from the core of the
product to its surface described by a pseudo-Fick law using apparent gas diffusivity. Diffusion of
chemical species in liquid phase is assumed negligible except for water where pseudo-Fick law is
used with an apparent liquid water diffusivity. In the open porosity of the product and at its surface,
the composition of the liquid phase and the gaseous phase in vapour and volatile compounds of
volatile chemical markers is assumed locally at equilibrium. Concerning the volatile chemical
markers, they are quantified using dynamic headspace-solid phase microextraction (HS-SPME) and
GC-MS system. The information given by this analysis is expressed in peak area on a
chromatogram and converted in gas concentration using a proportionality coefficient quantifying the
affinity of the species for the fibre selected for the extraction. For prediction of product physical
variables (temperature and moisture content) and composition in chemical markers (non-volatile
and volatile compounds), a relatively good agreement is found between predicted and experimental
measurements. Concerning stoechio-kinetic model, data about kinetic parameters for the apparent
reactional can not be found in the literature. A lack of data has also been noted for equilibrium data
for the volatile chemical markers: partition coefficient for the equilibrium between liquid and
gaseous phase and proportionality coefficient between the gaseous phase and the surface of the fibre
used for the HS-SPME measurements. For this reason, a trial and error methodology has been
adopted to find acceptable values for the kinetic parameters appearing in the stoechio-kinetic model.
These values must be considered as rough estimates (and hence used in extrapolation with caution)
taking into account the numerous hypotheses made when developing the model and the lack of data
in the literature for the model unknown parameters.

CONCLUSION
This work presents an attempt to predict mechanistically the extent of Maillard reaction during
baking of a sponge-cake type bakery product. The complexity of phenomena taken into account and
the lack of literature data about unknown model parameters force us to fix the optimal values of
unknown kinetic parameters using a trial and error methodology. The values of identified kinetic
parameters must hence be used with caution in extrapolated conditions.

REFERENCES
[1] Fehaili, S., Courel, M., Rega, B. & Giampaoli, P.. 2010. An instrumented oven for monitoring of
thermal reactions during the baking of sponge cake. Journal of Food Engineering, 101(3), 253-263.
[2] Rega, B., Guerard, A., Delarue, J., Maire, M. & Giampaoli, P. 2009. On-line dynamic HS-SPME for
monitoring endogenous aroma compounds realeased during the baking of a model cake. Food
Chemistry, 112(1), 9-17.

1868
 A methodology for the certification of food-serving services according to the

Mediterranean dietary model

Evangelos Grigoroudisa, Antonia Psaroudakib,c

a
Technical University of Crete, Chania, Greece (vangelis@ergasya.tuc.gr)
b
Technological Educational Institute of Crete, Sitia, Greece (psaroudaki@staff.teicrete.gr)
c
Agricultural University of Athens, Athens, Greece

INTRODUCTION
In the era of globalization, the concept of national dietary models has been remarkably reduced
to a minimum, given the fact that the selection of different types of food depends on more
factors compared to the past. Research and epidemiological results associating the
Mediterranean diet with good health have led to an examination of the economic aspect of the
Mediterranean diet. Certifying a dietary model presupposes its historic character and the exact
determination of its content. The aforementioned findings justify the need to certify this model
in food-serving businesses within the framework of a wider quality management system.
The characteristics of the Mediterranean diet are typical of the diet of Cretans in the 1960’s [1].
The term “traditional Mediterranean diet” is used to describe the dietary habits that were
typical in certain regions of the Mediterranean in the early 1960’s, such as Crete, some areas in
the rest of Greece and southern Italy. The association of the Mediterranean diet with good
health has boosted the desire of consumers, especially in tourist areas, to follow the
Mediterranean/Cretan dietary model [2]. This fact has led professionals in the food-serving
sector to advertise the delivery of such services.
The main objective of the paper is to investigate the potential of the Mediterranean dietary
model to inspire proposals of meals by food-serving services and suggests a methodology for
the certification of such services, according to this model. The development of the certification
procedure follows the structure of the ISO 9000 series.

MATERIALS & METHODS

For this study, we have used the results of the seven countries study on food types and general
dietary habits in Greece and areas of southern Italy in the 1960’s. The development of
specifications in the quality manual follows the Mediterranean diet pyramid, as it was
developed by the Oldways Health Organization, the World Health Organization and the
Harvard School of Public Health in 1942, taking into consideration dietary directions on food
expressed by the Supreme Special Scientific Health Council under the auspices of the Ministry
of Health and Solidarity [3].

RESULTS & DISCUSSION

The development of the certification procedure that follows the structures of the ISO 9000
series requires the compilation of quality manuals, including the assessment of quality policies,
the analysis of procedures, and the development of work instructions, as well as the
documentation of the whole quality management system [4], [5].

11th International Congress on Engineering and Food - Athens, Greece, 2011 1869
Food-serving businesses that wish to be certified based on the Mediterranean model should be
able to offer one or more Mediterranean menus. To achieve this, it is necessary to follow the
principles described in the production flow with reference to the selection, preparation and
serving of food according to the Mediterranean dietary model and to allocate responsibilities
and competencies so that the quality system can be applied and operate efficiently.
According to the seven countries study, the Mediterranean diet is characterized by [1]: higher
consumption ratio of monounsaturated fatty acids to saturated fatty acids, high consumption of
pulses, high consumption of grains (bread), high consumption of fruit, high consumption of
vegetables, moderate to high consumption of fish and seafood, moderate to high consumption
of milk and dairy products, low consumption of alcohol (wine) and low consumption of meat
and meat preparations. The above characteristics, translated into food quantities, are
represented in the food pyramid that was prepared by the Supreme Special Scientific Health
Council.
The main procedures followed by food-serving services are: (a) Production flow (supply and
selection of provisions, delivery of provisions and storage, preparation and cooking,
conservation and serving), (b) Compilation/Reorganization of menu, (c) Hiring and training of
employees, (d) Customer satisfaction measurement, and (e) Complaint management and
corrective actions.
The necessary documentation contains data and information that are critical for the justification
and the validity of the quality management system. The documentation is used as a proof that
the requirements of the quality management system are met. In general, the documentation
includes forms, checklists, records, and other lists (additional records that support specific
procedures).

CONCLUSION
In this study, we present the basic points for the compilation of a quality manual and a
procedures and directions manual, which can support the certification methodology of the
Mediterranean dietary model, following the structures of the ISO 9000 series. The benefits that
a business can earn from the certification of this dietary model and the wider system of quality
assurance of its services become apparent in a short time. A business that adopts such systems
becomes more competitive and its products and services become more useful, which results in
a remarkable boost in client confidence. At the same time, this contributes to the safeguarding
of an important cultural legacy, which can enhance the sustainability of the tourism sector.

REFERENCES
[1] Willett W.C., Sacks F., Trichopoulou A., Drescher G., Ferro-Luzzi A., Helsing E. & Trichopoulos
D. 1995. Mediterranean diet pyramid: A cultural model for healthy eating. American Journal of
Clinical Nutrition, 61, 1402S-1406S
[2] Trichopoulou A. & Lagiou P. 1997. Healthy traditional Mediterranean diet: An expression of
culture, history, and lifestyle. Nutrition Reviews, 55, 383-389.
[3] Special Scientific Health Council 1999. Dietary directions for adults in Greece, Archives of Hellenic
Medicine, 16(5), 516-524.
[4] Schlickman J. 2003. ISO 9001:2000 Quality Management System Design, Artech House, Norwood.
[5] Food Safety and Inspection Service 1997. Guidebook for the preparation of HACCP plans, US
Department of Agriculture, Washington.

1870
 Bactericidal effect of electrolyzed oxidizing (EO) water on E. coli O157:H7- and

Salmonella-inoculated beef, chicken, and shrimp

Jean Weese a and Tung-Shi Huang b

a
Poultry Science Department, Auburn University, Alabama USA (weesesj@auburn.edu)
b
Poultry Science Department, Auburn University, Alabama USA (huangtu@auburn.edu)

INTRODUCTION
This paper evaluates the bactericidal effect of electrolyzed oxidizing (EO) water on E. coli
O157:H7- and Salmonella-inoculated beef, chicken, and shrimp.

MATERIALS & METHODS

Strips of beef and chicken meat (25 g) were placed in a bacterial suspension of five-strain
mixtures of Escherichia coli O157:H7, Listeria monocytogenes, or Salmonella enteritidis for 2
min. After the strips were allowed to dry in the air for 10 min, they were treated with EO
water, aqueous chlorine, or tap water for 1, 3, 5 or 10 min. The removed strips were placed in
stomacher bags with 225 ml sterile Butterfield’s phosphate buffer and blended for 2 min at
normal speed. The suspension was serially diluted and 0.1 ml aliquots from appropriate
dilutions were spread onto the respective selective agar plates for each test bacteria. After
incubation at 37o C for 24 hr, colony numbers were recorded for the beef and chicken meat.
The shrimp were inoculated on day 0 and stored frozen at -20 °C. Bacterial enumeration was
done on day 0, 24, 49 and 119 of frozen storage.

RESULTS & DISCUSSION

S. enteritis, E. coli O157:H7, and L. monocytogenes on inoculated beef were reduced by 65.3-
83.6%, 60.9-75.4%, and 79.8-92.1%, respectively, and 87.1-93.6%, 80.6-94.9%, and 91.1-
97.6% respectively for treated chicken strips, following EO water treatment. No significant
difference in reducing bacterial loads was noted among the three treatment solutions.
However, a longer treatment time resulted in a greater bacterial loss.
Acidic EO water at 40 ppm free available chlorine was as effective as aqueous chlorine of the
same concentration, and significantly more effective (P < 0.05) than tap water in reducing
pathogen load on the inoculated shrimp. Further reduction of pathogen numbers was observed
following each frozen storage period. Pre-washing with alkaline EO water did not enhance the
bactericidal activity of the acidic EO water on the shrimp. The washed acidic EO water of the
inoculated shrimp had non-detectable bacterial populations when compared to treated aqueous
chlorine, alkaline EO water, and tap water

11th International Congress on Engineering and Food - Athens, Greece, 2011 1871
CONCLUSION
Meat texture and fat content may contribute to the greater bacterial loss in treated chicken meat
when compared to the beef. Acidic EO water can be used as an effective disinfectant to
replace aqueous chlorine for thawing shrimp blocks.

1872
 Predicting persimmon puree colour as a result of puree strength manipulation

Andrew R. Easta, Xiu Hua Tanb, Jantana Suntudproma

a
Institute of Food, Nutrition and Human Health, Massey University,
Private Bag 11 222, Palmerston North, New Zealand (a.r.east@massey.ac.nz)
b
Massey University (Singapore), Block T1A25, 500 Dover Road, Singapore

INTRODUCTION
Colour is an important quality defining attribute for any food product [1] as it is the first perceived
judgement of the quality of the food by the consumer. Quantitative measurement of the colour of foods is
routinely reported in the Commision Internationale de L’Eclairage (CIE) 3-dimensional model. As most
processed fruit products convert solid foods to liquid or semi-liquid products, the colour of the resulting
product is an important attribute in allowing the consumer to identify and establish authenticity that the
product is indeed made from the fresh fruit.
Non-astringent persimmon cultivars (e.g. ‘Fuyu’) ripen after harvest and can be consumed when either a
crisp apple like texture (firmness = 9 kgf) or a soft peach like texture (firmness = 1 kgf). During fruit
ripening, synthesis and degradation of pigments (carotenoids, chlorophyll and anthocyanins) occurs
resulting in dramatic colour changes. This natural change in colour due to ripening, creates challenges in
producing consistent colour properties of fruit based products. Pilando et al. [2] previously found that
strawberry maturity played an important role in influencing the colour characteristics of strawberry wine.
As solids concentration is likely to be manipulated and fruit maturity variable for any potential
persimmon fruit product, this work aimed to create a model that would enable prediction of the effect of
solids concentration manipulation on persimmon puree colour irrespective of fruit maturity.

MATERIALS & METHODS

Fruit were ripened at 20°C and destructively measured in order to identify approximately 7 kg of fruit
within three maturity classes representing firm (6.5-8 kgf), medium (3-6.5 kgf) and soft (0.1-3 kgf).
Persimmon purees were created by first preparing a large batch of the native state puree. This native state
puree was mixed in proportion with either de-ionised water or the 45% solids puree paste to create 10
purees in the range of half to double strength. Colour of the persimmon purees were measured
immediately after preparation. The experiment was conducted twice in order to create two sets of data,
one being used for model creation and the second for model validation.
A single model that was functional independent of fruit maturity was developed. All solids concentration
and colour parameter data (xi) was converted to a fractional change scale (¨x) relative to the values
measured for the native state puree (xn, Eq. 1). A simple second order polynomial (forcibly fitted through
the native puree characteristics) was used to fit and describe the patterns observed (Eq. 2) for each
CIELAB colour parameter. Validation of the resulting model was conducted by applying the equations
for predicting the colour change to the second independent data set. Model accuracy was quantified by
calculating the mean absolute error (MAE).
xi (1)
'x 1
xn

'x v 'S   z ( 'S ) 2 (2)

11th International Congress on Engineering and Food - Athens, Greece, 2011 1873
RESULTS & DISCUSSION
Irrespective of fruit maturity it was observed that dilution in the solids content of the puree result in
reduced L*, a*, b* and C parameters and increases in h, while the inverse was true for purees with
increased solids concentration (data not shown). These changes in the CIELAB colour parameters
suggest that dilution of solids content in persimmon puree results in a darker (lower L*), less intense
(lower C) and more yellow (increased h) product colour.
In most cases, the second order polynomial fitted through the native puree properties was found to be an
adequate description of the change in colour parameter as caused by a change in solids content (Table 1).

Table 1. Fitted constants for the second order polynomials to describe the fractional change in CIELAB colour
parameters as a function of the fractional change in puree solids content.
CIELAB Parameter v z R2
L* 0.139 -0.083 0.91
a* 0.943 -0.378 0.96
b* 0.651 -0.255 0.98
C 0.689 -0.273 0.99
H -0.113 0.122 0.89

Use of the polynomial prediction models found that the prediction was robust over a large range of
potential values (Figure 1). For the five CIELAB parameters the mean absolute error of prediction ranged
between 0.55 (for L*) and 1.17 (for h). The largest deviation from predictions occurred for purees made
from firm fruit and with increased (¨S > 0.5) solids concentrations. These deviations from prediction was
a result of low prediction of a* values from those observed (Figure 1b) which also results in prediction of
higher h values (Figure 1e).

Figure 1. Independent validation

(prediction vs observed) of use of
second order polynomial (Eq. 2; Table
1) to predict CIELAB colour parameters
(L*, a*, b*, C, and h) of persimmon
puree as a function of manipulation of
solids content.

CONCLUSION
Colour is an important attribute of fruit based products that can be substantially influenced by solids
concentration manipulation. In this work a second order polynomial was used to model the change in
CIELAB parameters with manipulation of concentration of solids in persimmon puree. The developed
models were independently validated and found to be robust, suggesting that prediction and hence
manipulation of colour outcomes is achievable through solids content manipulation.

REFERENCES

[1] Francis F.J. 1995. Quality as influenced by color. Food Quality and Preference, 6(3), 149-155.
[2] Pilando L.S., Wrolstad R.E., Heatherbell, D.A. 1985. Influence of fruit composition, maturity and mold
contamination on the color and appearance of strawberry wine, Journal of Food Science, 50, 1121-1125.

1874
 Occurrence of furan in commercial samples of roasted coffee in Brazil

Adriana P. Arissetoa, Eduardo Vicentea, Mariana S. Uenoa, Maria Cecília F. Toledoa

a
Institute of Food Technology, Campinas, Brazil (adriana.arisseto@ital.sp.gov.br)

INTRODUCTION
Furan is a food processing contaminant which occurs in several heat-treated foods, such as
canned and jarred foods, coffee and cereal products [1]. Furan is classified as a possible human
carcinogen and recent risk evaluations have indicated that the exposure to furan by commonly
consumed foods in the diet is a human health concern [2, 3]. Previous studies indicate that
roasted coffee contains the highest furan levels in comparison to other products, with mean and
maximum values of 1807 and 6900 ȝg/kg, respectively [4].
So far, no data on the level of furan in roasted coffee samples from Brazil is available in the
literature. Therefore, the objective of this study was to validate a method based on gas
chromatography coupled to mass spectrometry preceded by headspace solid phase
microextraction (HS-SPME-GC/MS) for furan determination and evaluate the levels of the
contaminant in roasted coffees available on the Brazilian market.

MATERIALS & METHODS

A total of 41 samples were purchased at supermarkets in the city of Campinas, SP, Brazil,
including traditional ground coffee of different intensities (n=27), instant (n=8), decaffeinated
(n=2) and premium coffee samples (n=4). All products were analyzed as bought. The SPME
was carried out by using a 75 ȝm carboxen-polydimethylsiloxane (CAR/PDMS) fiber, under
the previously optimized conditions, i.e. extraction temperature: 35ºC and extraction time: 30
minutes. Furan-d4 was used as internal standard. The method was validated according to the
guidelines laid down by the Brazilian Institute of Metrology, Standardization and Industrial
Quality [5].

RESULTS & DISCUSSION

In relation to the analytical method, good linearity over the range 0-9600 ȝg/kg was obtained
(r2 = 0.992). A comparison between curves set on aqueous standard solutions and on matrix by
applying the F-test and t-test revealed a non-significant matrix effect. Limit of detection (LOD)
and limit of quantitation (LOQ) were 3 and 10 ȝg/kg, respectively. Mean recoveries ranged
from 76 to 101%, and coefficients of variation ranged from 1.7 to 7.1% for repeatability and
from 6.2 to 13.8% for within-laboratory reproducibility.
The levels of furan in the analyzed samples are shown in Table 1. The furan content varied
from 250 to 5332 ȝg/kg. The lowest mean level was found in instant coffee (449 ± 271 μg/kg)
whereas the highest mean concentration was observed in strong ground coffee packed under
vacuum (4247 ± 1090 μg/kg). These results are comparable to data reported in the literature by
European and North-American countries. The high levels of furan found in roasted coffee may
be associated to the higher temperatures used in the roasting process, which exceed most of
other food processing procedures.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1875
 Table 1. Furan levels in roasted coffee
Product n Furan (μg/kg)
Mean Min-Max
Packaging in normal atmosphere
Classic 7 1670 1129-2026
Extra-strong 6 1556 1247-1861
Instant 8 449 250-1012
Packaging under vacuum
Classic 6 3472 2534-5021
Strong 2 4247 3340-5332
Extra-strong 6 2445 1556-5056
Decaffeinated 2 4082 3274-4778
Premium 4 1789 1273-2494

There was no correlation between furan levels and the coffee intensity as indicated on the label
(classic, strong and extra-strong). However, mean furan levels found in the samples packed
under vacuum were higher than those packed in normal atmosphere.

CONCLUSION
It is expected that these results will contribute to data accumulation for worldwide health risk
assessment and be helpful in establishing approaches to lower the exposure of the population to
furan from the consumption of coffee.

REFERENCES
[1] US FDA. Exploratory data on furan in food: individual food products; United States Food and Drug
Administration. 2004.
[2] IARC. Furan. In: IARC Monographs on the evaluation of carcinogenic risks of chemicals to humans;
International Agency for Research on Cancer. Lyon, v. 63, 1995.
[3] FAO/WHO. Summary and conclusions of the seventy-second JECFA meeting; Food and Agriculture
Organization/World Health Organization. 2010.
[4] EFSA. Update of results on the monitoring of furan levels in food; European Food Safety Authority.
2010.
[5] INMETRO. Orientação sobre validação de métodos de ensaios químicos DOQ-CGCRE-08. Instituto
Nacional de Metrologia, Normalização e Qualidade Industrial. Revisão 2, Junho 2007.

1876
 Potential of furan formation in roasted coffee as influenced by species and roast degree

Adriana P. Arissetoa, Eduardo Vicentea, Mariana S. Uenoa, Silvia A. V. Tfounia, Maria Cecília F. Toledoa

a
Institute of Food Technology, Campinas, Brazil (adriana.arisseto@ital.sp.gov.br)

INTRODUCTION
The findings of furan formation in heat-treated foods commonly consumed by the population
have attained worldwide concern since furan is classified as a “possible human carcinogen” [1,
2]. The pathways proposed to explain the furan formation in foods is mainly based on the
thermal degradation/Maillard reaction of sugars, alone or in the presence of amino acids,
thermal degradation of certain amino acids, and thermal oxidation of ascorbic acid and poly-
unsaturated fatty acids [3, 4]. It has been reported in the literature that roasted coffee contains
the highest furan levels when compared to other foods, with mean and maximum values of
1807 and 6900 ȝg/kg, respectively [5]. The levels of furan found in roasted coffee may be
associated to the higher temperatures used in the roasting process, which exceed most of other
food processing procedures.
Although there have been several studies on furan content in commercial samples of roasted
coffee, little knowledge is available on the relationship between roasting conditions and furan
formation. Thus, the objective of this work is to evaluate the influence of coffee species and
roast degree on furan levels in roasted ground coffee.

MATERIALS & METHODS

Green coffee beans of Coffea arabica (cv. Catuaí Amarelo IAC-62) and Coffea canephora (cv.
Apoatã IAC-2258) were roasted to light, medium and dark roast degrees, which were
determined by the Agtron/SCAA Roast Color Classification System. Roasted beans were
stored at -18ºC in aluminized valve bags and ground immediately before analysis.
The furan content was determined in the green and roasted ground coffees by using an in-house
validated method based on gas chromatography coupled to mass spectrometry preceded by
headspace solid phase microextraction (HS-SPME-GC/MS). The limits of detection and
quantification were 3 and 10 ȝg/kg, respectively.
Data were processed by analysis of variance one-way ANOVA and Tukey test for comparisons
of means (Statistica 5.5, Stat Soft Inc.).

RESULTS & DISCUSSION

Furan was not detected in green coffee whereas levels between 911 and 5852 ȝg/kg were found
in the roasted samples (n=20). This range is in accordance with data reported in the literature
[5].
Figure 1 shows the average furan levels for the species and roast degrees evaluated. When
considering the species, higher furan levels were observed in samples of C. canephora within
each roast degree. The higher levels of furan in C. canephora samples could not be explained
since it is not completely understood which components are the key reactants that could
contribute to relatively higher furan levels in some foods. In relation to the roasting process, it

11th International Congress on Engineering and Food - Athens, Greece, 2011 1877
was verified that furan levels significantly increased (p ” 0.05) with the intensity of roasting for
both species with darker roasted samples showing the highest levels of furan.

Different capital letters indicate statistic difference between species within a same roast degree.
Different small letters indicate statistic difference between roast degrees within a same species.

Figure 1. Mean furan levels in roasted coffee according to species and roast degree.

CONCLUSION
The results of the present study showed that the furan formation during roasting vary
depending on the coffee species and roast degree. The management of these parameters could
be taking into account as possible strategies to reduce the formation of this contaminant in
roasted coffee.

REFERENCES
[1] US FDA. Exploratory data on furan in food: individual food products; United States Food and Drug
Administration. 2004.
[2] IARC. Furan. In: IARC Monographs on the evaluation of carcinogenic risks of chemicals to humans;
International Agency for Research on Cancer. Lyon, v. 63, 1995.
[3] Locas C.P. & Yaylayan V.A. 2004. Origin and mechanistic pathways of formation of the parent furan
- a food toxicant. Journal of Agricultural and Food Chemistry, 52, 6830-6836.
[4] Becalski A. & Seaman S. 2005. Furan precursors in food: a model study and development of a simple
headspace method for determination of furan. Journal of AOAC International, 88, 102-106.
[5] EFSA. Update of results on the monitoring of furan levels in food; European Food Safety Authority.
2010.

1878
 Thermal inactivation of Byssochlamys nivea in pineapple juice combined with

preliminary high pressure treatments

Elisa Helena da Rocha Ferreiraa. Amauri Rosenthalb. Verônica Caladoa. Jorge Saraivac. Sónia Mendoc.
Pilar Rodrigues De Massaguerd

a
Federal University of Rio de Janeiro, School of Chemical Engineering, Rio de Janeiro. RJ. Brazil.
(elisahelenarocha@gmail.com)
b
Embrapa Agroindustria de Alimentos, Rio de Janeiro, Brazil. (arosent@ctaa.embrapa.br)
c
Aveiro University. Department of Chemistry. Aveiro. Portugal. (jsaraiva@dq.ua.pt)
c
Aveiro University. Department of Biology. Aveiro. Portugal. (smendo@ua.pt)
d
Fundação Tropical de Pesquisas e Tecnologia André Tosello. LABTERMO. Campinas. Brazil.
(pilar.rodriguez@terra.com.br)

INTRODUCTION
Byssochlamys nivea is a thermal resistant filamentous fungi and potential micotoxin producer.
Recent studies have verified the presence of ascospores of such microorganism in samples of
pineapple nectars and juices. Although the majority of filamentous fungi have limited heat
resistance and are easily destroyed by heat, Byssochlamys nivea ascospores have shown great
thermal resistance. The aim of this work was to evaluate the application of linear and Weibull
models on thermal inactivation (70, 80 and 90ºC) of Byssochlamys nivea ascospores in pineapple
juice after pretreatment with high pressure (550MPa or 650MPa during 15min).

MATERIALS & METHODS

For the preliminary high pressure treatment applied to the inoculated pineapple juice previously to
thermal treatment the following conditions were used: 550 e 650MPa for 15 minutes. Initial
temperature of high pressure treatment was set at 20 ºC. Samples inoculated with the mould were
inserted in sterilized polyethylene bags and pressurized. After on, the sample were transferred to
sterile eppendorf tubes and immersed in thermostatic baths, adjusted to the following temperatures:
70, 80 e 90ºC for 0, 5, 10, 15, 20 e 25 minutes. Following the thermal treatment, tubes containing
samples were immediately cooled down in ice bath and aseptically opened. Serial dilution and pour
plating were then carried out, using double concentrated Malt Extract Agar added with rose bengal
(0,25%), followed by homogenization. After mixture solidification, the plates were inoculated at
30ºC for 7 days. Analyses were done in duplicate.

RESULTS & DISCUSSION

Survival curves of B. nivea ascospores at 70°C after either pressure treatment and at 80°C after
550MPa for 15 minutes fitted well in both linear and Weibull models (Table 1). For the other
treatments, the Weibull model showed better fit. At 90ºC the Weibull model also showed a better
adjustment for ascospores inactivation (control) without previously high pressure treatment,
presenting a larger R2 and a smaller RMSE. In the others controls treatments (70 and 80ºC without
previously high pressure treatment), it was verified the activation of B. nivea ascospores, avoiding
the models adjustment. Table 2 shows the resulting parameters for Linear model (D-value) and for
Weibull (b e n) for each treatment applied to the mould.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1879
The parameter of form (n) varied proportionally with the temperature in both previous high pressure
treatment. D-value from Linear model was significant for all survival curves, while the parameters
of Weibull model were significant for some of the treatments. All parameters presented low
standard-deviation, assuring in that way the repeatability.
Previous pressure treatment at 650 MPa for 15 min. thermal resistance higher resistance to
inactivation was verified for the B. nivea ascospores inoculated in pineapple juice at 70 and 80ºC, or
possibly a higher capacity of the spores to adapt to the treatment (tail formation n  1). On the other
hand, in both treatments at 90ºC cumulative damage resulted in higher ascospores sensitiveness
(shoulder formation n ! 1).
By obtaining t1 value related to Weibull model as well as D-value associated to linear model (data
not shown), it was possible to design the thermal process required for each pressure with the
combination with each temperature considered. In this sense, after pressure application at
550MPa/15min. it would be necessary a thermal treatment of 16.50min. on the juice to obtain 5
logarithmic reductions in B. nivea ascospores population, while with preliminary treatment at
650MPa for 15min a heat treatment of 15.38min would be required. Preliminary high pressure
treatment contributed to ascospores inactivation at 90ºC and avoided activation in treatments at 70
and 80ºC.

Table 1. Determinant coefficients (R2) and residues square mean (MQE) from survival curve of B. nivea
ascospores inoculated in pineapple juice fitted using linear and Weibull models, after treatment at 550
MPa for 15 minutes and 650 MPa for 15 minutes

Ascospores of B. nivea in pineapple juice

After treatment at 550 MPa / 15 min After treatment at 650 MPa / 15 min
Weibull Model Linear Model Weibull Model Linear Model
R2 MQE R2 MQE R2 MQE R2 MQE

T*
70 0,99 6,4.10-4 0,99 7,9.10-4 0,87 0,0068 0,87 0,0069
80 0,94 0,90 0,91 1,39 0,93 0,43 0,93 0,44
90 0,99 0,12 0,91 1,11 0,99 8,3.10-6 0,95 0,16
*T = Temperature (ºC)

CONCLUSION
Preliminary pressure treatment contributed to B. nivea asospores inactivation in pineapple juice at
90ºC and avoided activation and 70 and 80ºC. Weibull model fitted better for the both applied
treatments. It was required 16.50min at 90ºC after treatment at 550MPa/15min and 15.38min after
650MPa/15 min., in order to obtain a 5 log-reduction, as recommended by the FDA [1]. However,
considering the long time required for inactivation which would imply in low process efficiency,
high energy demand an prejudice to quality characteristics such as sensory attributes, further studies
are necessary to improve the process aiming possible industrial application.

REFERENCES
[1] FDA. Food and Drug administration. 2001. Exemptions from the Warning Label Requirement for
Juice – Recommendations for Effectively Achieving a 5-Log Reduction. U.S. Food and Drus
Administration Centre of Food Safety and Applied Nutrition.

1880
 Role of spices on acrylamide formation in buckwheat ginger cakes

L. Markováa,b, Z. Ciesarováa, K. Kukurováa, H.ZieliĔskic, D.ZieliĔskad, A. Bednárikováa

a
VÚP Food Research Institute, Bratislava, Slovak Republic (markova@vup.sk, ciesarova@vup.sk,
kukurova@vup.sk)
b
VUT University of Technology, Faculty of Chemistry, Brno, Czech Republic (xcmarkova@fch.vutbr.cz)
c
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Olsztyn, Poland
(h.zielinski@pan.olsztyn.pl)
d
University of Warmia and Mazury, Olsztyn, Poland (dziel@uwm.edu.pl)

INTRODUCTION
Acrylamide is a probably probable carcinogen to humans, which is formed in foods during heat
treatment. The use of substances that allow effective elimination of acrylamide in foods [1] is
the effective way to reduce acrylamide content in food in order to protect human health.
According to our previous model study, the antioxidant capacity of some spice extracts highly
correlated with acrylamide formation in simplified simulated food matrices [2].
The aim of this study was to assess the impact of selected spices on acrylamide occurrence in
ginger cakes with the addition of buckwheat flour, which is frequently used as an additive in
functional foods due to its high level of antioxidants [3].

MATERIALS & METHODS

Samples of spices were separately extracted with mixture ethanol/water (1:1, v/v) and
antioxidant activity was determined by DPPH• radicals scavenging activity assay.
Dough for the production of ginger cakes consisted of light buckwheat flour and light rye flour
(in a ratio 0.7 : 0.3), buckwheat honey, raising agent, sugar, butter and spices (2 % content of
flour). Ginger cakes were baked at 180 °C for 18 min.
The extraction of acrylamide from the samples was implemented with CH3COOH (0.2 mmol/l)
and re-extraction into ethyl acetate. Acrylamide content was analysed by LC/ESI-MS-MS.

RESULTS & DISCUSSION

The acrylamide content in ginger cakes with spices (Figure 1) was compared with the values of
antioxidant activity of extracts from spices. The highest antioxidant capacity was determined in
extracts of cinnamon and star anise but the decrease of acrylamide content was not observed
for these two kinds of spices. The content of acrylamide in ginger cakes with star anise has not
changed. Acrylamide content in the ginger cakes with cinnamon increased even up to 29 %.
Conversely, acrylamide content was significantly reduced in ginger cakes with nutmeg, fennel,
anis and clove. A minimal decrease of acrylamide content was observed in ginger cakes with
vanilla, cardamom and white pepper. These kinds of spices showed much lower antioxidant
activity compared with cinnamon and star anise.
A content of acrylamide in ginger cakes could be influenced by chemical constituents with
antioxidant capacity of spices. Nevertheless, only weak correlation between results of

11th International Congress on Engineering and Food - Athens, Greece, 2011 1881
antioxidant capacity of spices extracts and acrylamide content of ginger cakes with correlation
coefficient of 0.68 was observed.
vanilla 227.14
cinnamon 333.60
allspice 266.27
nutmeg 197.26
coriander 306.00
clove 213.47
cardamom 234.87
fennel 202.47
white pepper 236.61
star anise 260.57
anise 212.61
without spices 258.22

0 50 100 150 200 250 300 350 400

Acrylamide [ȝg/kg]

Figure 1. Impact of selected kind of spices on acrylamide content in ginger cakes

CONCLUSION
The aim of this study was to determine the role of spices on acrylamide formation in
buckwheat ginger cakes. Acrylamide content was significantly reduced in ginger cakes with
nutmeg, fennel, anis and clove. In contrast, the highest antioxidant capacity was determined in
extracts of cinnamon and star anise. The change of acrylamide content was not observed in
ginger cakes with star anise. Acrylamide content in ginger cakes with cinnamon increased.
The final acrylamide content was probably influenced by compounds, which are found in
particular kinds of spices. The investigation of the effect of chemical constituents with
antioxidant capacity of spices is proposed in further studies using model systems.

ACKNOWLEDGEMENTS
This contribution is the result of the project implementation "The Centre of Excellence for
Contaminants and Microorganisms in Foods" supported by the Research & Development
Operational Programme funded by the ERDF. This work was also supported by the Slovak
Research and Development Agency under the contracts No. LPP 0310-09 and SK-PL-0051-09.

REFERENCES
[1] The CIAA Acrylamide "Toolbox". 2009. Confederation of the food and drink industries of the EU
[online]. [cit. 2011-02-02], 1-41. Available from: <http://www.ciaa.be>.
[2] Ciesarova, Z., Suhaj, M., & Horvátová, J. 2008. Correlation between acrylamide contents and
antioxidant capacities of spice extracts in a model potato matrix. Journal of Food and Nutrition
Research, 47(1), 1-5.
[3] ZieliĔski, H., Amigo-Benavent, M., Del Castillo, M.D., Horszwald, A., & ZieliĔska, D. 2010.
Formulation and baking process affect Maillard reaction development and antioxidant capacity of
ginger cakes. Journal of Food and Nutrition Research, 49(3), 140-148.

1882
 Detection of deoxynivalenol in wheat flour using fluorescence fingerprint

Junichi Sugiyamaa, Kaori Fujitaa, Mizuki Tsutaa, Masayo Kushiroa

a
National Food Research Institute, Tsukuba, Japan (sugiyama@affrc.go.jp)

INTRODUCTION
Deoxynivalenol(DON) is one of mycotoxins produced by Fusarium spp. in cone, wheat and
other grains. Conventionally, the gas chromatography-electron capture detector or the enzyme-
linked immunosorbent assay method has been used to detect DON, but these methods are time-
consuming, professional operation, and require expensive equipment and reagents. A new
method to detect DON quickly, easily, and accurately has been demanded.
The objective of this study is to detect DON in wheat flour using FF.

MATERIALS & METHODS

Samples
Japanese winter wheat variety “Hokushin” was used for this experiment. The wheat was
artificially infected with Fusarium graminearum s.str in an experimental field. Four levels of
contaminated wheat with different kinds of fungicide were harvested. They were named
Sample A, B, C and D from low to high contamination level.
Figure 1 shows preparation of wheat flour and experimental flow. After removal of foreign
substance from 120g of wheat grain for each sample, a cyclone sample mill (CSM-S1, UDY
Corp., USA) was used for milling to make wheat flour. Three sets of 20g sample were divided
into 15g for chemical analysis and 5g for FF measurement. The FF measurement was repeated
5 times for each 5g, so total of 15samples were measured in each contamination level. On the
other hand, the chemical analysis was carried out for 3 samples in each contamination level.

Chemical Analysis
Liquid Chromatography with UV detector (LC1100, Agilent Technologies, USA) was used to
measure the concentration of DON in flour [1]. ODS column (L-Column ODS,
4.6mmx250mm, 5Pm, CERI, Japan) was used. The wavelength of UV detector was set to
220nm.

FF measurement
A fluorescence spectrometer (F7000, Hitachi high technologies, Japan) was used for
measurement of FF[2]. The slit widths on both the excitation and emission sides are fixed at
10 nm. The interval of excitation and emission wavelength was fixed at 10 nm. The
wavelength scanning speed was set to 30 000 nm/min. The wavelength ranges were 200–900
nm for both excitation and emission. The charge voltage of photo multiplier was 350V.
Amount of flour sample for one measurement was 240mg.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1883
RESULTS & DISCUSSION
DON concentration varies from 2.4 ppm to 26.6 ppm. As a whole, it was relatively higher than
that observed in normal fields. This is because it was artificially infected with DON producing
bacteria. It was found that the sample A and B is almost the same concentration in average.
The concentration of the sample C was three times as much as that of the sample A and B. The
sample D was extremely contaminated and was about 10 times as much as the sample A and B.
PLS regression was applied to the same data as quantitative analysis. There were 60 samples
based on 4 contamination levels. We divided them into two even groups, which are calibration
dataset and validation dataset. PLS regression model was constructed in calibration dataset
with cross validation method. Six latent variables were adopted and the result was shown in
Figure 1. There were high correlation (R2=0.990) in calibration dataset. The validation dataset
had also similar correlation (R2=0.983). Because not much difference between SEC (1.1 ppm)
and SEP (1.4ppm) was observed, the prediction model was expected to be robust.

Figure 1. PLS regression of DON concentration.

CONCLUSION
Nowadays, modern information technology can handle this vast information at the same time.
The results of this research indicate that there will be high possibility to detect DON in wheat
flour non-destructively using FF.

REFERENCES
[1] Thammawong M., Okabe M., Kawasaki T., Nakagawa H., Nagashima H., Okadome H., Nakajima T.
& Kushiro M. 2010. Distribution of Deoxynivalenol and Nivalenol in Milling Fractions from
Fusarium-Infected Japanese Wheat Cultivars. J Food Prot., 73 (10), 1817-1823.
[2] Fujita K., Tsuta M., Kokawa M. & Sugiyama J. 2010. Detection of Deoxynivalenol Using
Fluorescence Excitation-Emission Matrix. Food Bioprocess Technol., 3 (6), 922-927.

1884
Modeling of growth and ochratoxin A production of Aspergillus carbonarius and evaluation in food

matrices: effect of (gel) microstructure, water activity, and temperature

A. E. Kapetanakoua, A. Abavia, S. Yanniotisb, E. H. Drosinosa, P. N. Skandamisa

a
Food Quality Control and Hygiene, Food Science & Technology, Agricultural University of Athens, Greece.
pskan@aua.gr
b
Food Process Engineering, Processing and Preservation of Agricultural Products, Food Science & Technology,
Agricultural University of Athens, Greece.

INTRODUCTION
In addition to the well-established effect of pH and water activity on microbial growth in foods, structural
properties (i.e., viscosity and liquid, semi-liquid or solid state) of foods are also known to highly affect
the probability and rate of microbial growth. Although the effect of food microstructure on microbial
growth has been studied extensively for bacteria and yeasts, limited information is available on how this
parameter affects fungal growth and mycotoxin production [1, 2].
The present study aimed to develop models for the combined effect of aw, microstructure (expressed as %
w/v gelatin), and temperature on growth and OTA production of A. carbonarius and to evaluate the
predictions of the developed models in food matrices of different viscosity such as custard, jelly, and
marmalade.

MATERIALS & METHODS

Growth and OTA production kinetics of A. carbonarius (ATHUM 5659) were determined in Malt Extract
Broth of aw 0.90, 0.94, 0.99 (by glycerol addition), supplemented with different gelatin concentrations (0,
5, 10 and 20%) at different temperatures (15, 20 and 25oC). Fungal growth and OTA production of A.
carbonarius (103 spores/mL) was also evaluated on food matrices of different viscosity, such as custard,
jelly and marmalade and incubated at 20oC. Fungal growth was estimated by measuring the dry fungal
biomass using sterile cellophane discs, while OTA quantification was performed by HPLC. The square
root of growth and OTA production rates were determined by the Baranyi model and were further
modeled as a function of temperature, gelatin concentration and aw by applying polynomial models.

RESULTS & DISCUSSION

The increase in gelatin concentration caused a significant decrease in growth and OTA production rates of
A. carbonarius at almost all examined assays (Fig. 1). Both models, for fungal growth and OTA
production, indicated that the influence of structure was less important as aw decreased and temperature
was not close to the optimum. The comparison of our results with previous reports was difficult due to the
limited number of studies on modeling the effect of microstructure on fungi; however our results may be
comparable to relevant models for structure on bacteria species [2]. Coefficients of determination were
0.90 and 0.89 for the models predicting the square root (¥ȝmax) of growth and OTA production rate,
respectively.
The mechanism through which microstructure affects the fungal growth and OTA production rates may
be associated with the mechanical distribution of water, diffusion of O2, nutrients or inhibitory
compounds, such as organic acids, and the mobility of microorganisms. Specifically, a number of studies
support that microorganisms in structured products, are immobilized and forced to grow in a restricted
space as colonies, posing an additional stress, which reduces their growth rate [2].

11th International Congress on Engineering and Food - Athens, Greece, 2011 1885
 Figure 1. Fitted curves of polynomial model for the square root of: (a) fungal growth (g-1) and; (b) OTA production
(ppm d-1) rates of A. carbonarius on laboratory media of different aw (0.99, 0.94 and 0.90) and microstructure (0, 5, 10,
and 20% gelatin) at 15 (___, Ŷ), 20 (___, Ÿ) and 25oC (___, Ɣ) in comparison with experimental data used for the
fitting. From: Kapetanakou et al. (2010). Food Microbiol (in press) doi:10.1016/j.fm.2010.06.001

Although a positive effect of glycerol on viscosity of culture media was observed (data not shown),
according to Table 1 the addition of glycerol did not significantly influence the performance of both
models. Predictions of fungal growth rate agreed well with the recorded data on custard and marmalade,
while predictions for OTA production rates showed low accuracy compared to observations in all studied
food matrices, except for marmalade (Table 1). Moreover, predictions for jelly from both developed
models indicated a poor correlation compared to the observed values. According to the manufacturer
company label, jelly powder contained ingredients of poor nutritional value, suggesting a possible cause
for the low recorded data compared the predictions. Another interpretation for the observed discrepancies
may be attributable to differences in intrinsic conditions (e.g., porosity and viscosity) and nutrients
between laboratory media and food matrices.

Table 1. Observed and predicted values of ¥ȝmax of fungal growth and OTA production on custard, jelly and
marmalade, incubated at 20ºC for 40-day of storage.
Fungal Growth Sq rate (g d-1) OTA Production Sq rate (ppm d-1)
Predictions Observations Predictions Observations
0.99aw 0.94aw 0.90aw 0.99aw 0.94aw 0.90aw
Custard 0.775 0.769 0.761 0.678±0.135 2.486 2.467 2.441 0.939±0.179
Jelly 0.731 0.725 0.718 0.312±0.082 2.320 2.301 2.276 0.193±0.049
Marmalade 0.402 0.399 0.396 0.339±0.188 1.129 1.118 1.102 0.711±0.212

CONCLUSIONS
The food structure markedly affects the growth and OTA production by A. carbonarius. The results of the
evaluation of model performance against different foods underlined the need to identify the critical
variables associated with food structure, which may account for the effect of the latter on microbial
behaviour and can be used in predictive models.

REFERENCES
[1] Brocklehurst, T. F, Mitchell, G. A. and Smith, A. C., 1997. A model experimental gel surface for the growth of
bacteria on foods. Food Microbiol. 14, 303–311. [2] Wilson, P.D.G., Brocklehurst, T.F., Arino, S., Thuault, D.,
Jakobsen, M., Lange, M., Farkas, J., Wimpenny, J.W.T. and Van Impe, J.F., 2002, Modelling microbial growth in
structured foods: towards a unified approach. Int. J Food Microb. 73, 275– 289.

1886
 Modelling of In-Mouth Perception The Case of Sodium

B.J.D. Le Révérend, I.T. Norton, S. Bakalis

School of Chemical Engineering, University of Birmingham

B15 2TT, Birmingham, United Kingdom (s.bakalis@bham.ac.uk)

INTRODUCTION
Over recent years, governmental and non-governmental health agencies have issued
recommendations aiming at reducing sodium intake. Although the recommendations are
targeting daily consumption 80% of the salt ingested by the consumer in the developed
countries comes from processed foods [1]. This puts pressure in food manufacturers to reduce
salt content without affecting sensory perception. It has been suggested that delivery profile
can be alter sensory perception [1]. In solid foods mass transfer and more specifically the rate
of mixing with saliva has been postulate to have a significant effect on salt perception.
However in liquid food matrices, mixing between the food and saliva is very efficient and in-
vivo measurements have shown a fast response to pulses of sodium in a solution delivered
using a gustometer or under sip-wise conditions.
The aim of this work is to develop a mathematical model that will be able to describe
phenomena occurring during oral processing with an emphasis to salt perception.

MATERIALS & METHODS

The model was built considering two main compartments:
1) a first representing the bulk of the mouth and is modelled as a stirred tank, using a
mass balance between food inlet (Vf, cf) and saliva (Vs, cs). Both fluids are mixed into a food-
saliva mixture (Vfs, cfs) according to a mixing time T which is a function of the inverse of the
viscosity of the food (Ș). This volume is emptied of 90% of its content (swallowing) when the
volume reaches 4 ml. The system of differential equations describing this mass balance was
solved using a forward Euler scheme.
2) a second is a diffusive volume that represents the mucus boundary layer close to
the vicinity of the tongue. This layer is of fixed thickness (e) and the diffusion of sodium ions
in this layer is that of sodium ions in water. Fickian diffusion through this layer was solved
using a 2nd order finite difference scheme.
Between those two compartments, a transfer coefficient h is controlling the exchange of mass
using a Neumann boundary condition Neumann boundary condition
h·(Cm|bulk-Cfs)=-D·(䳱Cm|bulk)·n.
This was estimated using a dimensionless analysis named Chilton-Colburn analogy that is
presented in eq. 1.

Sh = 0.0023 · Re4/5 · Sc1/3 (1)

where Sh is the Sherwood number Sh=h·l/D, Re is the Reynolds number Re=u·l·ȡ/Ș and Sc is
the Schmidt number Sc=Ș/(ȡ·D).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1887
The equations describing the system were solved using MATLAB (Mathworks, USA).

RESULTS & DISCUSSION

The proposed model compared favourably with experimental data as can be seen from Fig.1.
The model was then further developed to understand the effect of viscosity on the salt
perceived from foods having different release profiles. The results demonstrate that as viscosity
increases salt perception can be indeed modified by controlling the rate of delivery.

Figure 1. Comparison between the XRD measured and model prediction of the crystal structure of cocoa
butter in chocolate at different locations in the chocolate.

CONCLUSION
A conceptual and mathematical model describing the in-mouth momentum and mass transfer
during consumption of liquid was developed and tested against dynamic and static
experimental sensory data. The model was successful in predicting the dynamics of Time-
Intensity curves as well as overall saltiness perception in thickened salted solutions. The model
was then used to predict the effect of pulsed delivery of sodium in high viscosity liquids and
showed that it is a possible way to effectively reduce salt content without affecting saltiness
perception.

REFERENCES
[1] Drake, S. L., Lopetcharat K. & Drake M. A Salty taste in dairy foods: Can we reduce the salt?
Journal of Dairy Science 94, 636-645 (2011).

1888
 Furan derivatives dynamic in rye bread processing

V. Ozolinaa, D. Kunkulbergaa, B. Cieslakb, M. Obiedzinskib

a
Latvia University of Agriculture, Jelgava, Latvia (vro@apollo.lv)
b
Warsaw University of Life Sciences, Warsaw, Poland (beata_cieslak@sggw.pl )

INTRODUCTION
Furan is a volatile cyclic compound consisting of five members of the four CH groups and one
oxygen atom. Furans as well as a large proportion of aromatics form from the sugar
caramelization process at high temperatures or Maillard reaction at lower temperatures.
The IARC has evaluated furans as a possible carcinogen of the 2B group. Based on studies
conducted in laboratories with animals at high doses of furan, it is assumed that furan can
contribute to the incidence of cancer in long-term exposure to very low levels of furan in foods.
Furan compounds on heat–treated products can be found in the J. Maga study which gathered
data on their occurrences in food, their sensory properties and the formation pathways [1].
The European Food Safety Authority founded an evaluation group and during 2004-2009
collected data from EU Member States for furan content in food. Six Member States were
reported for furan content in grain products. There no such information from Latvia.
The FDA recommends HS-GC/MS methodology for furan determination, where furans are
analyzed as volatile compounds. The method represents a new approach to the rapid
characterization of food products. High resolution mass spectrometry allows information to be
gathered on molecular ion origin, creating a permanent database of the product compounds
from the volatile part of sample composition. Temperature, equilibrium time and the vial size
are key parameters to be optimized for each food [2].
The aim of the present study was to investigate formation of furan and its derivatives in
Latvian whole-grain rye bread crust and crumb during baking

MATERIALS & METHODS

The samples used in this experiment were obtained from rye whole-grain flour (stock company
‘Jelgavas Dzirnavas’, Latvia).The dough was made with scald, natural starter and sugar, salt,
malt and cumin. The dough sample was fast frozen to stop fermentation and sublimated in
vacuum drying equipment. The whole-meal rye bread was baked in a Latvian commercial
bakery in a wood-fired oven 30, 45, 60 and 75minutes.
After baking, the loaves were left to cool at room temperature for 12 hours. Crumb and crust of
bread loafs were examined on presence of furan and derivatives. Solid phase micro extraction
(SPME) was applied for sampling and analysis of furan by means gas chromatography-mass
spectrometry (HS-SPME-GC/MS).
Quantitative determination were made using internal standard addition of 1,2-dichlorobenzen
and simultaneous scanning mode (mass range 38-200 m/z) and selected ion monitoring (SIM)
of ions (m/z) 39 and 68 for furan and respectively m/z 146 for IS.
The experiment was carried out in the Research Laboratory of Department of Biotechnology,
Microbiology and Food Valuation, Warsaw University of Life Sciences in 2010.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1889
RESULTS & DISCUSSION
Nine furan derivatives were identified in Latvian baked rye bread: 1) 2-Furancarboxaldehyde;
2) Ethanone, 1-(2-furanyl)-; 3) 2-Furancarboxaldehyde, 5-methyl-; 4) 2-Furanmethanol; 5) 2-
Furancarboxaldehyde, 5-(hydroxymethyl)-; 6) Furan, 2-pentyl-; 7) 2-Furanmethanol, acetate;
8) 1-(2-Furanyl)-2-hydroxyethanone; 9) 5-Formyl-2-furfurylmethanoate.
The presence of furans has been ascertained in rye bread crumb already at 30 minute of baking.
Rapid formation of 2-Furancarboxaldehyde (furfural) was observed in the subsequent baking
stage, and at 45 minutes at a temperature of 99 ºC reached maximum value in comparison with
IS. The furfural amount in the next 15 minutes of baking decreases dramatically, and continues
to decline after the optimal baking time of 60 minutes. In 1993, Silwar and Lullmann observed
a similar dynamic in the process of green coffee roasting where furfural fully formed after 5
minutes at 230 ºC, then fell rapidly, and decomposed at a higher temperature. In 1963 Smith
found that furfural forms from the oxidation of furfuryl alcohol and is also formed by
decomposition of pentosans, for instance by dehydration of arabinose. In 1991 Mottram
showed that furfural is formed from the Amadori compound of a pentose and an intermediate
3-deoxyosone. [3].
In this study it was observed that 2-Furancarboxaldehyde, 5-methyl- formation is taking place
at an early stage of the baking process, reaching a maximum at 45 minutes of baking at 98 ºC,
then decreases until the end of baking. In 1969 Fagerson described that 2-
Furancarboxaldehyde, 5-methyl-forms in the thermal degradation of glucose [3].
Other derivatives - 2-Furanmethanol, Ethanone, 1-(2-furanyl) and 5-HMF in rye bread crumb
formed in negligible quantities at the early stage of baking with maximum reached at 45
minutes, than decreases until the end of baking.
During baking temperature increases rapidly in bread crust to higher than 100 ºC. Nine furan
derivatives were found in rye bread crust, four derivatives more than in rye bread crumb. In
addition to the crumb, identified derivatives Furan, 2-pentyl-; 2-Furanmethanol, acetate; 1-(2-
Furanyl)-2-hydroxyethanone; 5-Formyl-2-furfurylmethanoate were detected in more quantity
in rye bread crust.
Comparing the formation of furfural in rye bread crumb and crust, it is seen that the amounts at
45 minutes of baking are significantly different.

CONCLUSION
Nine volatile furan derivatives were identified in Latvian rye bread- 5 in bread crumb and 9 in
bread crust. The majority of the volatile furan derivatives form at early stage of baking, and
further reduce during baking. During baking the 2-Furancarboxaldehyde, 5-methyl-(5HMF)
formed in negligible amounts in Latvian rye bread crust but increased with prolonged baking
time.

REFERENCES
[1] Maga J.A. 1979. Furans in foods. Critical Reviews in Food Science and Nutrition, 11, 355-400.
[2] Altaki M.S., Santos F.J. & Galceran M.T. 2007. Analysis of furan in foods by headspace solid-phase
microextraction-gas chromatography-ion trap mass spectrometry. Journal of Chromatography A,
1146, 103-109.
[3] Flament I. 2002. Coffee Flavor Chemistry, John Willey and Sons, England, 410 p.

1890
 The effects of Heracleum Platytaenium Boiss Essential Oil on The Growth of

Ochratoxigenic Penicillium Verrucosum (D-99756) Isolated From Kashar Cheese

Sibel Ozcakmak a, Muhammet Dervisoglub, Abdullah Akgunc, Adnan Akcind,

Tülay Aytaú Akcin e, Fatih Seyisf

a
Department of Food Processing, Terme Vocational School, Ondokuz Mayis University, Samsun, Turkey
(sibelo@omu.edu.tr)
b
Department of Food Engineering, Engineering Faculty, Ondokuz Mayis University Samsun, Turkey
(mderviso@omu.edu.tr)
c
Department of Food Engineering, Engineering Faculty, Trakya Univerisity, Edirne, Turkey
d
Biological Science Department, Faculty of Science and Literature, Amasya University, Amasya, Turkey
e
Biological Science Department, Faculty of Science and Literature, Ondokuz Mayis University, Samsun,
Turkey
f
Field Crops Department, Faculty of Agriculture, Bozok University, Yozgat, Turkey

INTRODUCTION
Penicillium species are frequent contaminants of different food products and are known to
produce a variety of secondary metabolites. P. verucosum is primarily encountered on cereals
and cheeses [1]. Various species of Heracleum have been reported to possess antibacterial and
antifungal properties [2].

MATERIALS & METHODS

Materials
Heracleum platytaenium Boiss essential oil was obtained from Biological Science Department,
Faculty of Science and Literature, Amasya University, Samsun-Turkey. Penicillium
verrucosum Dierck (D-99756), obtained from Faculty of Food Engineering in Istanbul
Technical University, Istanbul-Turkey. Yeast Extract Sucrose (YES) agar, YES broth,
methanol, Tween-80, Sodium Clorid (NaCl) were obtained from Merck, Germany.

Methods
Minimal inhibitory concentrations (MIC) and minimal lethal concentrations (MLC) were
evaluated using the method of Tavares et al. (2008) [3]. The fungal growth was indicated by the
turbidity. The highest dilution (lowest concentration), showing no visible growth compared
with Eo-free controls, was regarded as MIC. The Eo dilution, which is not visible growth of
typical P. verrucosum colonies on the plates, was accepted as lethal effect. The MLC was
defined as the lowest Eo concentration at which 99% of the inoculums was killed. The
inoculations of 1 mL for pour plate method and 30 μL for three point inoculation method from
the tubes were sub cultured on YES agar plates and than incubated at 25oC for 72 hours. All the
experiments were performed in duplicate and three independent experiments were run with
concordant results.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1891
RESULTS & DISCUSSION
The mycelia growth was visible in negative control tube while any fungal growth wasn’t seen in
positive control tubes. MIC and MLC values were at 31 and 125 μL mL-1, respectively.
Determined MLC values were the same in both of two assays. The inhibitory effect of spice
essential oils is mainly due to the most abundant component. Heracleum species has just been
investigated for anticandidal activity due to the presence of myristicin, elemicin and (E)-
anethole by some researches [2, 4]. This study demonstrated that H. platytaenium oils had the
most fungistatic and fungicidal affect on ochratoxigenic P. verrucosum.

REFERENCES
[1] Larsen T.O., Svendsen A. and Smedsgaard J., 2001. Biochemical characterization of ochratoxin
producing strains of the genus Penicillium. Appl. Environ. Microbiol., 67, 3630-3635.
[2] Kuljanabhagavad1 T., Sriubolmas N. and Ruangrungsi1 N., 2010. Chemical composition and
antÕmicrobial activity of the essentÕal oil from Heracleum sÕamÕcum. J Health Res., 24(2), 55-60.
[3] Tavares AC., Gonçalves MJ., Cavaleiro C., Cruz MT., Lopez MC., Cahoto J. and Salgueir LR., 2008.
Essential oil of Daucus carota subsp. halophilus: Composition, antifungal and cytotoxicity. J of
Ethnopharmacology, 119, 129-134.
[4] Iscan G., Ozek T., Ozek G., Duran A. and Baser KHC., 2004. Essential oils of three species of
Heracleum anticandidal activity. Chemistry of Natural Compounds, 40(6), 544–547.

1892
 The Inhibition of Contaminated Molds by Some Essential Oils in Cheeses

Sibel Ozcakmak*a, Abdullah Akgunb, Muhammet Dervisogluc

a
Department of Food Processing, Terme Vocational School, Ondokuz Mayis University, Samsun, Turkey
(sibelo@omu.edu.tr)
b
Department of Food Engineering, Engineering Faculty, Trakya Univerisity, Edirne, Turkey
(abdullahakgun@yahoo.com)
c
Department of Food Engineering, Engineering Faculty, Ondokuz Mayis University, Samsun, Turkey

INTRODUCTION
Mould growth on cheeses should be inhibited by some preservation methods, such as chemical
food preservatives. But consumer perception that the use of industrially synthesized food
preservatives may be associated with potential toxicological problems has generated interest in
the use of naturally occurring compounds [1]. Some certain herbs, spices or their oils with well-
known antimicrobial properties have been used for a long time in some foods such as cheese to
prevent fungal growth [2].

THE CONTAMINATED FUNGI STRAINS COMMONLY ISOLATED FROM

CHEESES
Mold growth except mould-ripened cheese varieties on the cheese surface causes undesirable
flavor, economic loses and quality problems which of them are capable of producing toxic
secondary metabolites. The genus most frequently isolated was Penicillium sp. producing
mycotoxins such as ochratoxin-A and citrinin as responsible for spoilage in cheeses [3].

USING SOME EOS FOR THE CONTROL OF UNDESIRED MOLDS IN CHEESES

The selected plant essential oils have been classified as generally recognized as safe (GRAS) by
the Unites States Food and Drug Administration (FDA) as approved flavors or food additives
[4]. It was determinded that the growth of Penicillium sp. could be inhibited with phenolic
compunds Eos [5].

CONCLUSION
Natural phenolic compounds could inhibit the growth of the fungi and their toxin production.
This is a consumer pressure to reduce the use of such preservatives and perhaps replace them
with other more natural ones. They could be used by the food industry as antifungal agents
without toxic risk. Eos or their products could be treated with cheeses in order to protect from
fungal contamination. The using of some essential oils treatment on the surface of cheeses for
inhibiting undesired fungi may be an alternative protection instead of chemical in future.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1893
REFERENCES
[1]. Leung AY. 1978. Encyclopedia of Common Natural Ingredients Used in Food, Drugs and
Cosmetics, 309–311, Wiley, New York.
[2]. Coma V. 2008. Bioactive packing technologies for extend shelf life of meat-based products. Meat
Sci 78, 90-103.
[3]. Torkar KG, Vengust A. 2008. The presence of yeasts, moulds and aflatoxin M1 in raw milk and
cheese in Slovenia. Food Control 19: 570-577.
[4]. Burt S. 2004. Essential oils: their antibacterial properties and potential applications in foods- A
review. Int J Food Microbiol 94: 223-253.
[5]. Vazquez BI, Fente C, Franco CM, Va´zquez MJ, Cepeda A. 2001. Inhibitory effects of eugenol
and thymol on Penicillium citrinum strains in culture media and cheese. Int J Food Microbiol 67:
157–163.

1894
 Fungicidal against Aspergillus flavus and Decontaminate AflatoxinB1 with

Neutralized and Acidic electrolyzed oxidizing water

Li Lite, Xiong Ke

College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, P.R.China

ABSTRACT
One of serious food safety problems is likely to be Food-borne mycotoxins. Some mycotoxins
such as aflatoxins (especially aflatoxinsB1) mainly produced by A.flavus, are considered to be
serious contaminant of food. This study investigated the effectiveness of Neutralized
electrolyzed oxidizing water (NEW) and Acidic electrolyzed oxidizing water (AcEW) on
Fungicidal against Aspergillus flavus and decontaminate AflatoxinB1 (AFB1).
Although the results reveal both NEW and AcEW possessed potent to inhibit A.flavus, they
have different fungicidal efficiency on A.flavus. Different .OH level in NEW and AcEW
accounts for this different. AcEW and NEW also have the potent to decontaminate the AFB1
produced by A.flavus. The main conversion production produced by AFB1 treatment with
AcEW and NEW were isolated and identified. It is showed that AFB1 (C17H12O6) produce the
8-Cl-9-OH-AFB1 (C17H13O7Cl) treatment with AcEW and NEW. The conversion produce of 8-
Cl-9-OH-AFB1 belongs to amphiphilic molecules, which different from the fat solution of
AFB1. Furthermore, the mutagenicity assay showed that the conversion produce did not exhibit
mutagenic activity.

KEY WORD: Neutralized electrolyzed oxidizing water (NEW); Acidic electrolyzed oxidizing
water (AcEW); OH radical (.OH); Conversion structure; Mutagenic responses

11th International Congress on Engineering and Food - Athens, Greece, 2011 1895
1896
 Influence of different inulin types on bread quality in the process of freezing and thawing

J. S. Filipoviüa, Ĉ. B. Psodorova, N. K. Filipoviüb, V. S. Filipoviüc

a
Jelena Filipoviü Ph.D, Institute for Food Technology in Novi Sad Bulevar cara Lazara 1, 21000 Novi Sad,
Serbia (jelena.filipovic@fins.uns.ac.rs)
a
Ĉorÿe Psodorov Ph.D, Institute for Food Technology in Novi Sad Bulevar cara Lazara 1, 21000 Novi Sad,
Serbia(djordje.psodorov@fins.uns.ac.rs)
b
Nada Filipoviü Ph.D, Faculty of Technology, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia(nfil@uns.ac.rs)
c
Vladimir Filipoviü Bs., Mlinpek Instiute, Bul. Oslobodjenja 66b, 21000 Novi Sad, Serbia
(filipovic@mlinpekzavod.com)

ABSTRACT
Bakery products, particularly bread, are constituent parts of meals in all dietary patterns. By
enriching the composition of bread with fibers, recognized as, prebiotic substances, long term
health benefits can be easily met. Two different types of commercial inulin were incorporated into
the dough formula as flour supplements at the level of 5 %. Dough freezing/thawing kinetics was
determined at -18°C and +20°C, respectively (the highest determination coefficient is experienced
with 5% of inulin GR 0.948 and 0.982, and the lowest with the dough without fibers 0.915 and
0.976).The dough was frozen at -18°C and stored over a period 30 days. The results concerning
bread quality changes were interpreted through bread with volume and crumb quality.

CONCLUSIONS
The following conclusions can be drawn:
Inulin type, regarding the length of side molecular chains, has the greatest influence on the freezing
and thawing kinetics and quality of final product.
Freezing and thawing kinetics can be reliably described by square polynomial (Y= b0+b1*X
+b2*X2) profile for unsteady temperature change with introduced value of t2 referring to the phase
transformation, thus enabling computing the process.
The highest positive effect on freezing/thawing kinetics, confirmed by high determination
coefficient R2 is experienced with 5 % of inulin GR (0.948 and 0.982, respectively).
Over a longer period of storing inulin with longer molecular chains is beneficial in preserving the
quality of final products at the initial level and the adverse effect of fiber on bread crumb quality is
not evident.
In the period of 30 days, doughs without and with inulin GR, deterioration of the quality of final
products is similar, thus pointing at inconvenience of this type of inulin in the frozen dough
technology.
Bakery products enriched with inulin HPX can be successfully produced by frozen dough
technology, enabling quick preparation of a wider variety of fresh and appealing final products and
distributing to larger areas, i.e. to consumers with special nutritive needs.


11th International Congress on Engineering and Food - Athens, Greece, 2011 1897
ACKNOWLEDGMENTS
These results are part of the project supported by the Ministry of Science of the Republic of Serbia


1898
 Thermal analysis of strawberry preservation by cooling and freezing

Adrian-Gabriel Ghiausa, Catalina Vasilescua

a
Technical University of Civil Engineering, Bucharest, Romania (ghiaus@instal.utcb.ro)

INTRODUCTION
Healthy diet for the population requires large consumption of fresh fruit and vegetables
throughout the year. Their seasonal and perishable features, especially in the case of
strawberries, make them difficult to commercialize outside the harvest period. Therefore,
different conservation methods and techniques are used aiming to maintain, as much as
possible, the properties of the fresh product and to achieve a high quality final product for the
out of the season. Cooling and freezing are the most common conservation methods used for
strawberries to enjoy their sweet taste all year round.
The objective of this study is to investigate the mechanisms through which different parameters
affect the rate and uniformity of cooling. The thermal behavior of the strawberries is analyzed
depending on the heat transfer mechanism used for cooling process.

MATERIALS & METHODS

The time depending temperature field inside the strawberry was numerically simulated using
the COMSOL Multiphysics commercial software package which is based on finite element
method. The Heat Transfer module was used to simulate different ways of heat transfer, e.g.
conduction, convection and radiation as well as combination between them. Boundary
conditions were imposed for each specific surface of the berry according to the analyzed
cooling system. A non-uniform mesh consisting of 2076 triangles elements has been generated.
The temperature distribution in the product is calculated with the differential equation of the
transient heat conduction [1]:
wt k
’ 2t (1)
wW U cp
where t is the temperature, W is the time, k is thermal conductivity, U is the density and c p
is the specific heat. The solution of this differential equation is found with the finite element
method.
Heat transfer from the product surface to the air of the cooling room is made by natural or
forced convection and it is based on the Newton law:

q hcv t p  t f  (2)
where q is the heat flux, hcv is the convection heat transfer coefficient, t p is the temperature
of the product surface and t f is the temperature of the cooling air.

RESULTS & DISCUSSION

Initially, the product is at the temperature of 20 °C and it is introduced in a cooling room where
the air temperature is maintained constant at the value of 0 °C using a refrigerant that is

11th International Congress on Engineering and Food - Athens, Greece, 2011 1899
evaporated. It is assumed that the product is homogenous and the thermophysical properties are
constant: thermal conductivity, k is 1.1 W/m·K, density, ȡ is 800 kg/m3 and the specific heat, cp
is 4000 J/kg·K [2].
The product is progressive cooled from the surface to the interior of the product. Inside the
product, the heat is transferred by conduction and the heat is released at the surface of the
product with natural convection.

Figure 1. Simulated time temperature profile

One of the cases, which are analyzed, is the cooling by the natural convection with the ambient
air that has the temperature of 0 ºC. The product is placed on a support made by a good
insulation material and no heat transfer occurs between the support and the product. The
convection heat transfer coefficient is assumed to be equal to 5 W/m2K. Figure 1 presents the
temperature distribution in the cooled product after 4 hours and 10 minutes and the variation of
the temperature during the cooling process in two characteristics points, A and B: at the surface
of the top of the product and in the middle of the product, respectively.
The analysis of the temperature distribution at different time intervals leads to the optimization
of the operating parameters and the choice of the cooling system in order to obtain high quality
cooled product.

CONCLUSION
A simulation of the cooling process of strawberry was performed with the software package
COMSOL Multiphysics that is based on finite element method. The cooling of the product was
studied when different heat transfer mechanisms are used: conduction and convection. For the
presented cooling case with natural convection, the temperature on the surface is lower at the
top and increases slightly to the bottom.

REFERENCES
[1] Ghiaus A.-G., 2003, Heat Transfer, Editura Conspress, Bucharest
[2] ASHRAE, 2006, Refrigeration
[3] Ghiaus A.-G., Vasilescu C., 2010, The analysis of strawberries freezing methods and systems with
numeric simulations, The Conference of the Building Services Faculty, 18-19 March

1900
 Effects on Xe hydrate formation for texture in vegetable tissue

Hiroko Andoa, Toru Suzuki b, Kazuhito Kajiwara a, Yoshinori Kawagoe c, Yoshio Makino c, Seiichi
Oshita c

a
School of Bioscience and Biotechnology, Tokyo university of Technology, Tokyo, Japan
(hiando@bs.teu.ac.jp)
b
Department of Food Science and Technology, Tokyo University of Marine Science and Technology,
Tokyo, Japan (toru@kaiyodai.ac.jp)
c
Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan
(aoshita@mail.ecc.u-tokyo.ac.jp)

INTRODUCTION
Texture of “fresh” vegetable tissue degrades significantly by freeze-thawing treatment. It has
been attempted to improve the freezing technique in order to prevent such texture degradation
after thawing. On the other hand, it was reported that the storage time of vegetable after harvest
could be extended when it was storied in xenon gas (Xe) atmosphere at 5 oC, where the water
in the vegetable is just before formation of gas hydrate, then forms so called structured liquid
water [1]. Recently, we proposed an interested storage technique using Xe hydrate formation
for fresh vegetable tissue, instead of freezing. Gas hydrates are crystalline solids composed of
water and gas as Xe. It was confirmed that Xe hydrate can form in onion tissue by pressurizing
between 0.4 and 0.8 MPa Xe pressure at 5 oC. Additionally, it was found that the formation
ratio of Xe hydrate in the tissue can be arbitrarily determined by controlling the initial pressure
[2]. As a next step to find a suitable storage condition, this study investigated the effect of Xe
hydrate formation on texture in onion tissue by varying the hydrate formation ratio. Especially,
two texture parameters of fracture stress and initial modulus in onion tissue after the
decomposition were used to evaluate the change of texture

MATERIALS & METHODS

Soft core of a fresh onion was cut into a 4 x 4 x 10 mm3. This tissue sample was pressurized
between 0.4 and 0.8 MPa Xe pressure at 5 oC for the Xe hydrate formation ratio (gas hydrate/
whole free water) to be 15~25, 26~35, 36~45, and 46~55 %. The formation ratio in the tissue
was confirmed by NMR measurement with solid echo pulse sequence (MU25A, JEOL, Japan).
As a reference, N2 gas also induced to the samples under same condition. After that, fracture
stress and initial modulus as texture characteristics of onion tissue after gas hydrate
decomposition were measured by using texture analyzer (RE2-33005S, Yamadam co., ltd.,
Japan) at room temperature.

RESULTS & DISCUSSION

Figure.1 shows change of fracture stress (a) and initial modulus (b) in onion tissue after Xe
hydrate formation and decomposition with increasing the hydrate formation ratio. Both
parameters after decomposition were lower relatively than that of fresh tissue though it is not
shown in Fig.1. Furthermore, as shown by white keys in figure.1 (Ƒ, ż), when it was stored in
0.8 MPa N2 gas pressure at 5 oC for 1 week, instead of Xe, the texture degradation in the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1901
onion tissue was not observed because water did not crystallize. It was found that texture for
onion tissue degraded some extent by Xe hydrate formation, however, the pattern of the change
with the hydrate formation ratio shows a significant difference between texture characteristics.
Fracture stress was kept a constant value (61 x 104 N/m2, approximately) which was
independent of the hydrate formation ratio even though it decreased a little. On the other hand,
the initial modulus decreased linearly with increasing the hydrate formation ratio. Previously, it
was reported that initial modulus might be a texture characteristic related to cell membrane
water permeability in vegetable tissue. On the other hand, fracture stress reflects cell wall
strength in the tissue. [3]. Thus, it was considered that the cell membrane of onion was more
sensitive to Xe hydrate formation rather than the cell wall. Particularly, the damage degree of
cell membrane changed gradually depending on the hydrate formation ratio.
Both fracture stress and initial modulus in vegetable tissue after freeze-thawing usually
decreased dramatically than that after Xe hydrate formation and decomposition. It is a key
point how to prevent the damage for cell membrane in order to keep vegetable tissue texture
after freeze-thawing. On the other hand, by using Xe hydrate instead of freezing, it is possible
to prevent the damage for cell membrane with the hydrate formation by controlling the hydrate
formation ratio.

100 160
(a)
90 (b)
140
Fracture stress, 104 N/m2

80
Initial modulus, 104 N/m2

120
70
60 100

50 80
R2 = 0.96
40
60
30
40
20
10 20

0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Gas hydrate formation ration in onion tissue, %
  Gas hydrate formation ration in onion tissue, %

  Figure 1. Change of texture, fracture stress (a) and initial modulus (b), depending on the increase of
Xe hydrate formation ratio in onion tissue. Error bar shows upper and lower values (n=3).

CONCLUSION
It was considered that using of Xe hydrate was expected as a novel preservation technique for
fresh vegetable instead of freezing. Particularly, it would be an useful technique to prevent the
damage of “cell membrane” in the tissue by controlling the hydrate formation ratio.

REFERENCES
[1] Makino et al., Agricultural Engineering International: the CIGR Ejournal, VIII (June), 1, 2007.
[2] Ando et al., 5th International Technical Symposium on Food Processing㸪Monitoring Technology in
Bioprocesses and Food Quality Management㸪1㸪Germany㸪08㺃2009㸬
[3] Ando et al., Food Preservation Science, 34, 261, 2008. In Japanese.

1902
 The potential of ambient cooling systems for reducing refrigeration loads and saving

energy

Stephen J James, Christian James

Food Refrigeration and Process Engineering Research Centre (FRPERC), The Grimsby Institute of
Further & Higher Education (GIFHE), HSI Building, Origin Way, Europarc, Grimsby, North East
Lincolnshire, DN37 9TZ, UK (jamess@grimsby.ac.uk)

INTRODUCTION
The surface temperature of cooked products is very high when they leave baking ovens or deep
fat fryers, and consequently the difference between the surfaces and the ambient is very large at
that time. To reduce energy usage, and costs, a number of food manufacturers have
traditionally operated two-stage cooling operations using ambient air followed by refrigerated
air. An initial higher temperature stage can also be used to improve the energy efficiency of
freezing operations. With many ambient stable products such as bread and confectionary
filtered ambient air is a common cooling method after the product exit backing ovens.
However, the use of ambient cooling is not widespread within the food industry and in some
cases it is not encouraged. This study investigates the ambient cooling of hash browns prior to
freezing, and the ambient cooling of meat and vegetable pies prior to blast chilling.

MATERIALS & METHODS

Hash brown freezing. Targeted trials were carried out to determine the amount of heat and
moisture that could economically be removed from fried hash browns using ambient cooling
prior to freezing. Batches of fried hash browns were cooled under a range of air velocities
followed by freezing in air at -30°C and 3.5 ms-1.
Pie chilling. Trials were carried out to produce an efficient energy saving process that could be
installed after the company moved to new premises. Batches of cooked pies were cooled under
different combinations of air temperature and air speed using air at -10°C or 0°C (±0.3°C) and
three air velocity settings.

RESULTS & DISCUSSION

Table 1. Core temperatures in hash browns during ambient (22°C) cooling
Air Core temperature (°C)
velocity
Time (min)
(ms-1)
0 2.5 5 7.5 10 12.5 15 17.5 20
2.0 86.8 70.1 56.9 47.2 40.2 34.9 31.0 28.3 26.1
2.5 76.0 66.0 55.3 47.0 40.2 35.0 31.3 28.4 26.2
3.0 83.1 70.9 57.3 46.9 39.4 34.0 30.1 27.3 25.3

Hash brown freezing. In the initial ambient cooling trials the core temperatures of the hash
browns were 40±1°C after 10 minutes at all three air velocities (Table 1). After a further 10
minutes the product core temperatures were approaching (within 2.5°C) that of the ambient

11th International Congress on Engineering and Food - Athens, Greece, 2011 1903
temperature. During the first 5 minutes of ambient cooling the hash browns lost approximately
1 g of weight and less than one third of a g in the following 5 minutes (Table 2).

Table 2. Weight loss from hash browns during ambient cooling

Weight loss (g) during different time (min) periods
During
0 to 5 5 to 10 10 to 15 15 to 20 20 to 25 25 to 30
frying
Mean (SD) 2.25 (0.30) 0.94 (0.07) 0.31 (0.03) 0.22 (0.05) 0.13 (0.02) 0.15 (0.05) 0.12 (0.02)

Pie chilling. In the pie cooling, trials using a 30 minute pre-chilling ambient (20°C) cooling
stage before chilling at 0°C, showed that the core temperature of the pie could substantially be
reduced by this process (Table 3). The overall cooling time to 5°C was increased but the time
required in the blast chiller slightly reduced as was the heat extraction required by the blast
chiller.

Table 1. Pie temperatures after 30 minutes, total and time in chiller to reach 3°C. Control is the time
achieved in a single stage system at 0°C, 3 ms-1
Conditions Pie temperature (°C) Time to 5°C (min)
after 30 min Total In Chiller Control
30 min at 20°C then 0°C on grid, both 64 140 110 117
3.3ms-1
30 min at 20°C then 0°C on tray, both 60 120 90 100
3.3ms-1
Time to 3°C (min)
30 min at 10°C, 1.3 ms-1 then 0°C, 3.3ms-1 45 136 106 120
on tray
30 min at 10°C, 3.3 ms-1 then 0°C, 3.3ms-1 47 136 106 120
on tray

CONCLUSION
The introduction of an ambient cooling operation prior to refrigerated chilling or freezing
operations can significantly reduce the heat load and energy consumption of chilling and
freezing systems. Two cases studies have been presented that clearly demonstrate this.
Hash browns emerged from a fryer at 80°C and had to be frozen to -12°C before packaging at a
process rate 4.5 tones/h. The existing spiral freezer was incapable of extracting the initial heat
load and the moisture loss from the hash browns was causing ice to build up the evaporator.
An initial 5 minutes of ambient cooling removed 562,500 kJ of heat energy from the 4.5 tonnes
of hash browns every hour. It also prevented 60 kg per hour of water freezing on the evaporator
In the pie case, by combining experimental studies with modelled data a two-stage process was
designed incorporating an initial ambient cooling stage followed by a redesigned multi-rack
chilling tunnel. This work showed that approximately 50% of the heat that had to be extracted
in the total cooling pie cooling process could be removed in a 30 minute ambient pre-chilling
stage. This two-stage system has now been installed by the company and shown to achieve
more controlled, uniform and faster cooling of the pies than previously achieved with a single
chiller.

1904
 Industrial Superchilling, A Practical Approach

A.M. Stevika, I.C. Claussenb

a
SINTEF Energy research, Kolbjørn Hejes vei 1D, NO-7465 Trondheim, Norway
(astrid.stevik@sintef.no)/ (ingrid.c.claussen@sintef.no)

INTRODUCTION
An industrial, automated superchilling process line needs to combine the requirement for
gentle handling of a valuable raw material with demands for energy efficiency, high capacity
and product quality. The results from the study of three alterative process lines for superchilled
processing of fresh cod showed large variations with respect to processing time, need for
manual operations, skinning errors and yield. The processing time and need for operators was
very high for the Marel superchilling concept, while the quality and yield was high. The
alternative involving skinning in a Baader59 unit followed by superchilling in an impingement
freezer was very rapid and showed competitive results for quality and yield. Superchilling in
the impingement freezer followed by skinning in the Baade59 unit resulted in a substantial
amount of skinning errors in spite of very rapid processing.
An efficient process presupposes a high yield combined with efficient production, large
capacity, few operators, few skinning errors and an end product of high quality. The results
from the current experiments indicate that superchilled processing of cod by means of skinning
in the Baader59 unit followed by superchilling in the Impingement freezer would be the most
competitive alternative, taken the above mentioned factors into account. The method implies
short processing time, low amount of skinning errors, little manual operation as well as high
yield and quality.
Superchilling is a robust method for conserving fresh food, and brings along many possibilities
due to the extended product shelf-life. In the near future it is considered likely that most
industrial processing of fresh fish will be done by means of superchilling, and the pioneer
industry currently taking on the method will have a great advantage based on the experience
from final development and implementation of the superchilling technology.
The results discussed in this paper are based on a series of tests performed in November 2010
at the site of a large Norwegian fish producer.

MATERIALS & METHODS

Three alternatives for superchilled processing of Atlantic cod (Ghadus morhua) were tested as
shown in Figure 1; (1) the Marel Superchiller concept process line, (2) Superchilling of cod
fillets in an Impingement Advantec Lab Freezer, and (3) Skinning of the cod fillets in the
Baader59 unit followed by superchilling in the JBT Impingement freezer .
The experiments were performed on 1-3 kg cod fillets, caught 4-6 days ahead of the
experiments. For all process alternatives the cod was pre-chilled in ice-slurry at -1,2 ºC, filleted
in Baader 184/182 filleting machines for white fish before the further processing were split in
the three mentioned alternatives. Quality assessment of the fillets was performed shortly after
processing of superchilled fillet, and six days after temperature equalization.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1905
 Superchill Skinning
Alt.1 IceͲ Slurry
CBC Skaginn

PreͲ Filleting Superchill Skinning

Alt.2
cooling Baader 184/182 Impingement Baader59

Skinning Superchill
Alt.3
Baader59 Impingement

Figure 1. Experimental setup

RESULTS & DISCUSSION

Processing through alternative 1 induced major bottlenecks in the production line due to loss of
orientation of the fillets when entering the salt brine. This also generated a need for many
operators upfront the CBC- superchilling unit. The second and third alternatives were both very
rapid and generated no comparable need for manual operations. However, processing by
alternative 2 induced a great amount of skinning errors, and for alternative three the skinned
fillets easily stuck onto the Impingement transport band during superchilling.
The yield and share for fresh end product showed little variation for the three alternatives,
favouring alternative 1 by a few percent (3,7% higher share for fresh end product than for
Alternative 3). This result is not adjusted for the quality loss due to freezing of the fillets unto
the Impingement transport band.

CONCLUSION
Based on the results above superchilled processing of cod is considered most feasible by means
of alternative 1 or alternative 3. An efficient process presupposes a high yield combined with
efficient production, large capacity, few operators, few skinning errors and an end product of
high quality. Of the three alternatives tested, processing by means of skinning of filets in the
Baader59 unit followed by superchilling in the impingement freezer seems to be the most
promising alternative due to the short processing time, low amount of skinning errors, the need
for few operators as well as high yield and quality. The method presupposes that the
superchilling step can be handled automatically, and that the fillets don’t stick unto the
transport band during chilling.
Superchilling is a robust method for conserving fresh food, and brings along many possibilities
due to the extended product shelf-life. This involves i.e. increased production capacity,
simplified production planning as well as opening up for new kinds of product and the ability
to reach new markets. Automatization is very important also for the fish industry and
superchilling as a rapid and repeatable production method is very suitable for automated
processing. In the near future it is considered likely that most industrial processing of fresh fish
will be done by means of superchilling, and the pioneer industry currently taking on the
method will have a great advantage based on the experience from final development and
implementation of the superchilling technology.

1906
 Evaluation of thermal resistance and efficiency of palm olein and canola oils in frying of

potato chips

Shadi. Bolouriana, Ali. Rafeb , Gholamali. Goli Movahheda,b, Majid. Afsharia,b

a
Department of food additives, Iranian Academic Center for Education Culture and Research
(ACECR), Mashhad, Iran, P.O.Box: 91775-1376, ( shadibolourian@yahoo.com)
b
Department of Food Science and Technology, Ferdowsi University of Mashhad (FUM),
PO Box: 91775-1163, Mashhad, Iran (alirafe1400@yahoo.com)

ABSTRACT
Frying is a popular way for food processing. Selection of suitable oil is an important factor that
affects the quality of fried foods. This study was carried out in order to optimize the blend of
palm olein and canola oils as frying oil. At first, various compositions of these oils were
prepared and their heat resistance was evaluated in 120°C using rancimat method. The oil
blends were used for frying of potato in order to evaluate their frying performance. Frying was
carried out for 5 successive days in 180°C and changes in total polar compounds (TPC), acidity
and peroxide values of oils were determined. Results were shown that TPC, acidity and
peroxide value increased during frying. Increase in canola ratio increased the peroxide value of
used oil. In fact, a blend of palm olein and canola oils with ratio of 9:1 was chosen as a best
formula. It had highest induction time (18.72h) in rancimat method and it's TPC after frying
(17.72%) was lowest.

CONCLUSIONS
Although there was some differences in the trend of quality of formulated oil by using of the
measured parameters, but it may be possible to propose that the F2 formula is suitable oil. This
formula was advocate the highest time of induction period in the method of rancimate and the
total production of polar compound was less. In addition to, the oil uptake of potato chips was
less than the other formula (i.e. 31.8%). In spite of; the peroxide value was higher than the F0
and F1. By mixing of olive oil and palm olein, the stability of oil was increased. The
comparsion of stability oxidative was indicated that F2 oil was better than the commercial
control oil (table 4).

Table 4. Comparison of commercial parameters of control oil with F2 oil

specification Oil F2
Induction period at 120 °C (houre) 18.72 13.46
total polar compounds 17.72 22.37
peroxide value 5.88 5.12
Acidity 0.12 0.08

11th International Congress on Engineering and Food - Athens, Greece, 2011 1907
1908
 Assessment of furfurol derivatives: food risk factors in natural apricot and peach juice

Jianu Călina, Cocan Ileanaa, Jianu Ionela

a
Banat`s University of Agricultural Sciences and Veterinary Medicine, Faculty of Food Technology Products,
Timisoara, Romania (negreaileana@yahoo.com)

INTRODUCTION
Expression „protective foods” (Mincu, I., et al. 1989; Segal, B., et al, 1999, Durlach, J. Et al, 1999)
includes foods rich in bioactive compounds („protective factors”) obtained from the vegetable, fruit.
Natural juices, apricot / peach („liquid fruit”) by their nutrient / biologically active native potential are
functional foods (protective) with health benefits (hyperacidity, diabetes, gout, senescence,
cardiovascular disease, etc).
Their processing technologies include inevitably thermal unit operations (peeling, monitored enzyme
inactivation, softening texture, chicken) (figure 3) requiring technological steam.
Furfural (F) (2 – furaldehyde, furfurol) (CAS 98-01-1) and some of its derivatives [5-(hydroxymethyl)
furfural (5-HMF) (CAS 67-47-0) heterocyclic carbonyl compounds is commonly found in varying
amounts in various foods rich in carbohydrates (hexose) as a result of improperly monitored thermal
processing which hexose respectively pentose its dehydrate the acid catalysis.

MATERIALS & METHODS

Colorimetric determination of azomethine compounds of F Юi 5-HMF
It can perform in liquid phase (apricot and / or peaches juice) used as reference in identical cells that in
the gas phase (bubbling atmosphere in the processing space through a trap) (figure 3). In both cases
proceed classic by calibration curve plotting to 5-HMF reference, în intervalul 10 – 100 Ȗ/mL. Linearity
range Lambert-Beer is 0-5Ȗ/mL. Absorption maximals recommended for colorimetric assessment 580
and 640 nm. Color reagent solution its prepare with validity checked of 24 h, by dissolving 0,375 g
diphenylamine (CAS 122-39-4) analytical purity in 15 mL glacial acetic acid (CAS 64-19-7) and 9 mL
concentrated hydrochloric acid. Can be diluted depending on the emissions concentration of 5-HMF
during procedure. The maximum error recorded is ±1%, reproducibility ± 0,341%. In determining were
no reported other interference.

RESULTS & DISCUSSION

Preliminary research monitored the changes (variation) of the organic composition of the apricots and
peaches throughout the determined duration of the vegetation period (growth, maturity) subsequently
stored until usage. The three geographical area (AG1, AG2; AG3) of reference for the studied fruits
belong to the Western Plain of Romania (Northerns latitude 45°; 22´ respectively Eastern longitude
21°; 25´) with a predominantly chernozemic soil leached with humus, next to halomorphic,
hydromorphic, meadows and sands. The moderate plain climate with oceanic and sub Mediterranean
influences registers an average temperature of 21 to 23°C (the hottest month and rainfall between 600 and

1
11th International Congress on Engineering and Food - Athens, Greece, 2011 1909

700 mm and predominantly Western winds). The complex phenomenon of growth and maturity of the
apples includes a chain of biochemical processes that generate and determine the accumulation of
carbohydrates in ratios specific to the species and breed following their utilisation in other metabolic
processes. In our research, fructose, glucose and sucrose during a growing cycle of apricots and peaches
have overall slight accumulation. Thus the glucose for peach oscillates between Astfel 10-20 mg/edible
part (in cuticle) and between 20 – 30 mg/edible part (in pulp). Weight ratio of fructose / glucose
remained constant (1,1/1). During maturation (months V,VI, VII for apricots, respectively VII, VIII, IX
for peachs, sucrose remains constant (5,1 – 5,3 g/100 g edible part). During storage (depositing) to
processing total carbohydrate content is changed for both types (between 9,6 – 13,8% for apricots
respectively between 6,3 – 12,8% for peaches). For storage temperatures of 20-22°C total carbohydrate
loss by respiration is between the 1 – 1,8% for apricots and between 1 – 1,7% for peaches. Storage in
controlled atmosphere (94% CO2/ 6% O2) significantly decreased weight loss (between 0,7 – 0,8%) for
glucose, fructose and sucrose. Lower organic acids and mono-polycarboxylic from apricots and peaches
addition to the major role of intermediaries in the overall metabolism, in sensory qualities determine the
equilibrium acid - base of the system. Amount of their share in the free state and combined (total acidity)
in studied case, with titratable acidity expressed as mg/% edible part confirmed for apricots (malic acid
1000, citric acid 400) and for peaches (malic acid 340, citric acid 250) values are consistent with
dissociation constants for malic acid (K1=3,86·10-4; K2=1,29·10-5) and for citric acid (K1=8,7·10-4;
K2=1,8·10-5; K3=4,0·10-6). Malic and citric acids evaluated were 95,08% of total water soluble organic
acids. They but in smaller proportion (below 250 mg/% edible part) and ascorbic (K1=7,94·10-5;
K2=1,62·10-12), succinic (K1=6,3·10-5; K2= 3,4·10-16), tartaric (K1=1,04·10-3; K2=4,55·10-5), lactic (K1 =
1,4 · 10-4) acids are responsible for the thermal dehydration of glucose (5,36%), fructose (7,54%) and
sucrose (after split in fructose and ‫܈‬i glucose (5,34%)). Dynamics of titratable acidity proved to be
proportional to the storage conditions for apricots (1,51% initially, respectively 1,32% after 5 days at
22°C storage and 1,51% initially, respectively 1,49% after 10 days at 3°C). For peaches (0,79% initially,
respectively 0,68% after 30 days at 0 - 1°C and 0,79% initially, respectively 0,73% after 63 days at 0°C
in Controlled-atmosphere (with 5% CO2; 3%O2; 92%N2). In the same storage period for all apple types
that are evaluated the process of forming and transforming protopectine in soluble pectin, associated
with the reduction of the total pectin quantity. Simultaneously after 60 days of apple storage at 4°C and
80% relative humidity the texture firmness decreases by 3,0 kgf (piston penetrometer with a 6 mm
diameter).

CONCLUSION
Continuous Colorimetric Monitoring of furfural and their its major derivatives can be considered a
preventive method available, fast, secure of risk warning, adventitious presence of these compounds in
natural juices of apricots and/or peaches as a result of „aggressive” thermal processing of primary fruits.

REFERENCES
[1] Mincu, I. ‫܈‬.a., 1989, Orientări actuale în nutri‫܊‬ie, Ed. Medicală, Bucure‫܈‬ti.
[2] Segal, B., Segal, R., 1991, Tehnologia produselor alimentare de panifica‫܊‬ie, Ed. Ceres, Bucure‫܈‬ti.
[3] Jianu, I., Delia Dumbravă, 2000, Factori de protec‫܊‬ie alimentari, Ed. Mirton, Timi‫܈‬oara.

1910 2

 Numerical evaluation of liquid food heat sterilization in a brick-shaped package

Pedro E. D. AUGUSTOa; Marcelo CRISTIANINIb

a
Department of Food Technology (DTA), School of Food Engineering (FEA), University of Campinas
(UNICAMP); Technical School of Campinas (COTUCA), University of Campinas (UNICAMP),
Campinas, SP, Brazil (pedro@cotuca.unicamp.br)
b
Department of Food Technology (DTA), School of Food Engineering (FEA), University of Campinas
(UNICAMP), Campinas, SP, Brazil (olecram@fea.unicamp.br)

INTRODUCTION
Thermal processing is one of the most utilized methods for food preservation. Appertization is
still the most effective conservation method, even when compared to recently advanced
techniques.
The liquid flow characteristics inside the packaging during heating are a function of its
geometry, where even small alterations can change the process characteristics. However, little
attention has been deposited on modifications in the thermal processing of liquid foods through
changes in geometry or orientation of its packaging [1, 2].
The present work aimed to evaluate the thermal process of a low viscosity liquid food in a
brick shaped package, as the influence of its orientation on the process lethality.

MATERIALS & METHODS

Simulations were performed by CFD analysis. The three-dimensional model was obtained
from the actual geometry of the brick shaped package, a retortable multilayer carton-based
package. The unstructured tetrahedral mesh was generated based on previous work.
Due to the small thickness of the packaging, its thermal resistance was considered negligible.
Water was considered as a liquid model food, and its thermal properties were used as function
of temperature. The initial conditions generally used in the literature were considered. As
boundary condition, heating and cooling were considered to be uniform, with heat flow
obtained at each time step by the retort convective heat transfer coefficient (h) and temperature
(Th), based on previous work. Minimization of RMS was used as a criterion of convergence.
The time step used was 2.0 s. Efficiency of the thermal process was compared for three
possible package orientation, based on mass average sterilization value (Fm; Equation 1; Tref =
121.1 ºC, zClostridium botulinum = 10 ºC).
V tf T t ,V Tref
1 n
V V³0 t³0
Fm 10 z
dtdV J process ˜ DT
ref
Equation 1

RESULTS & DISCUSSION

The temperature and velocity profiles were compatible with those described in different works
heating different products in cans and bottles, as in the pasteurization of water in bottles,
sterilization of water in cylindrical cans, pasteurization of water in cylindrical cans,
sterilization of viscous fluid in cylindrical cans and bottles.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1911
In the beginning of heating, there is the presence and disintegration of various Benard Cells,
circular flows formed due to the meeting of two or more flows, as the package heating is
uniform through its sides. If heating is continuous, the flow tend to stabilize in a characteristic
profile, with a larger looping ascending in package walls and descending in the center of the
package, and small ones near the center of the package base, at an opposed direction to the
first. Fluid flow took almost 1000 s to stabilize, for the three orientations. The same behavior is
then observed during the beginning of cooling.
The fluid flow points down to a coldest region, called the slowest heating zone (SHZ), in
contrast with the cold spot characteristic of conductive products heating. The fluid temperature
is distributed in layers, with the hot ascending flow close to the package walls during heating.
The SHZ is located in the bottom of the package during the process.
Packaging orientation was evaluated due to the big differentiation of its dimensions (94 mm of
height, 83 mm of depth, and 45 mm of width), which results in a difference of one order of
magnitude in Grashof number (Gr) and Rayleigh number (Ra).
Orientation does not result in different sterilization values (Fm). This behaviour is similar to
water pasteurization in beer cans [3], but contrary to the sterilization of consistent liquid foods
in conical [1, 2] and cylindrical [4] cans.
This difference can be attributed to the convection flow inside packaging, which is high
influenced by its geometry and by the evaluation itself. In Varma and Kannan [1, 2] and Ghani
et al. [4] works, the authors evaluated the orientation only by temperature profiles. Although
comparison among the efficiencies in the thermal processes must be done through sterilization
values, this approach is rarely adopted. We highlight once more that liquid food thermal
process efficiency must also be evaluated by its mass average sterilization value (Fm).

CONCLUSION
The present work has shown for the first time the internal thermal and velocity profiles of
liquid foods thermally processed in a brick shaped package, as well as its sterilization values
due to processing. The results obtained demonstrated the potential of using computational fluid
dynamics (CFD) in evaluating thermal processes of liquid foods, especially in new or non-
conventional packaging geometries. It can be concluded that packaging orientation does not
result in different sterilization values during thermal process of water in the brick shaped
package.

REFERENCES
[1] Varma, M. N. and Kannan, A. (2005). Enhanced food sterilization through inclination of the container
walls and geometry modifications. International Journal of Heat and Mass Transfer, 48(18), 3753-
3762.
[2] Varma, M. N. and Kannan, A. (2006). CFD studies on natural convective heating of canned food in
conical and cylindrical containers. Journal of Food Engineering, 77(4), 1024-1023.
[3] Augusto, P. E. D.; Pinheiro, T. F. and Cristianini, M. (2010). Using computational fluid-dynamics
(CFD) on the evaluation of beer pasteurization: effect of can orientation. Ciência e Tecnologia de
Alimentos, 30(4).
[4] Ghani, A. G. A.; Farid, M. M. and Chen, X. (2002). Numerical simulation of transient temperature
and velocity profiles in a horizontal can during sterilization using computational fluid dynamics.
Journal of Food Engineering,51(1), 77-83.

1912
 Effect of steam jet cooking on the destruction of corn starches

L.H. Ferng,* S.H. Chen, Y.A. Lin

Department of Food Science, National I-lan University, I-lan City, Taiwan (lhferng@niu.edu.tw)

INTRODUCTION
Aqueous starch dispersions have many practical applications in food products. Steam jet cooking
has been used for years to prepare aqueous starch dispersions for food and non-food application.[1][3]
This technique involves pumping starch slurry through an orifice and mixing with steam at high
temperature and pressure. It is well known that the steam jet cooking introducing high turbulence
and large pressure drop generated high shear stress to starch.[2] There is limited information about
the destruction of starch granules by steam jet cooking. The objective of this research is to study the
effect of shear stress on structure of corn starch granules by steam jet cooking.

MATERIALS & METHODS

A laboratory scale steam jet cooker has been established with flow rate about 1L/min and cooking
temperature up to 145qC. Three kinds of corn starch, waxy, regular, and high amylose were used.
Starch slurries (5% w/w) were cooked by steam jet cooker at temperature 100qC (SJ100), 120qC
(SJ120) and 135qC (SJ135) compared with hot water boiling at 90qC 30 min. (HB). The cooked
dispersions were rapidly cooled by liquid nitrogen, dried by freeze drying, ground, and sieved
through 80 mesh. The cooked starches were washed with water and centrifuged to obtain the
insoluble particles. The particles were investigated by particle size analyzer, scanning electron
microscope (SEM) and damage starch assay kit.

RESULTS & DISCUSSION

To understand the effect of steam jet cooking on integrity of starch particles, the raw and cooked
starches were washed three times with cold water, the insoluble particles were isolated by
centrifugation in centrifuge at 2000g for 20 min. Percent yield was determined by freeze drying the
water-washed particles. The percent yields of insoluble particles of raw starches were all excess 98
%. The effect of heat treatment was revealed by decreasing in percent yield of insoluble particles.
There were also significant differences in percent yield of insoluble particles among different
starches. There was a significant decrease in percent yield of insoluble particles of cooked
dispersions for all three starches in the order HB > SJ100 > SJ120 > SJ 135. That is the starch
granule destruction was exacerbated by jet cooking and increasing temperature. The percent yield
of insoluble particles of waxy corn decreased from 56.2 for hot water boiling to 18.8 for steam jet
cooking at 120qC, regular corn from 81.7 to 75.9, and high amylose corn from 96.9 to 93.9. The
data also showed that SJ100 has higher destruction than HB, although the temperature of heat
treatment was similar. This could be due to the shear effect of steam jet cooking.
To understand the size of insoluble particles, the dried sample was dispersed in cool water and the
particle distribution was analyzed by particle size analyzer. The results of particle size distribution
analysis revealed that the insoluble particles swelled on contact with water, the particle size of
studied starches were much larger than uncooked raw starches, and the particle size of HB was
larger then SJ100 for all starches. This may be due to the time of cooking, 30 min. for hot water
boiling vs. a few sec for steam jet cooking at 100qC.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1913
To investigate the damage of cooked starch particles, damage starch assay kit was used to test thr
percentage of damaged starch. Percentage damaged starch of cooked dispersions for all three
starches became higher with increasing of cooking temperature.(Table 1) The high amylose corn
starch(Hylon VII) was more resist to steam jet cooking than the others. Although, the time of heat
treatment was much shorter for steam jet cooking at 100qC than hot water boiling, the percentage of
damaged starch was higher. This also revealed the effect of shear stress from steam cooking.
On SEM observation, waxy corn starch granules appeared to be fractured by all heat treatments.
The damaged granules showed sponge like structure for the starch dispersions of normal corn starch
cooked by hot water boiling. The starches heated by steam jet cooking were fractured into small
fragments. For high amylose corn starch, hot water boiling treatment only caused granule swelled.
Steam jet cooking at 100qC not only swelled the granule but also fractured some filamentous pieces
from starch granule. When cooking temperature increased high than 120qC, the starch granules
were fractured and sponge like structures were observed.

Table 1. Percentage damaged starch of waxy, normal corn starch and Hylon VII after hot water boiling
and steam jet-cooking.
Starch damage (%)
Sample
Hylon VII Normal corn Waxy corn
Untreated 1.91 ± 0.02e 1.06 ± 0.01c 1.22 ± 0.02e
Boiling 36.49 ± 0.27d 71.90 ± 0.18b 74.56 ± 0.53d
c b
SJC-100 38.99 ± 0.27 72.01 ± 0.24 77.53 ± 0.11c
b b
SJC-120 40.06 ± 0.51 77.07 ± 0.93 79.81 ± 0.86b
a a
SJC-135 41.11 ± 0.11 80.60 ± 0.06 83.19 ± 0.79a
Reported values are the mean ± SD (n = 3).
a-e
Values in a column for each sample with different superscripts are significantly different (p
< 0.05).

CONCLUSION
The effect of shear force by steam jet cooking on destruction of corn starches granule is significant.
The comparison between hot water boiling and steam jet cooking at 100qC revealed the destruction
of shear force by steam jet cooking. The starches heated by steam jet cooking were fractured into
small fragments. The degree of destruction relates to the amylose content of corn starch. Waxy
corn with little amylose is easy to swell during heat treatment and to destroy by shear force of jet
cooking. High amylose corn starch (Hylon VII) sustained the integrity of starch granule after heat
treatment of hot water boiling and steam jet cooking at 100qC. The study offered basic information
of starch granule destruction by steam jet cooking for further application in food product
development and processing design.

REFERENCES
[1] Klein, R.E. & Brogly, D.A. 1981.. Method for selecting the optimum starch binder preparation system.
Pulp and papers 55:98-103.
[2] Fanta GE. & Eskins K. 1995. Stable starch-lipid compositions prepared by steam jet cooking.
Carbohydrate Polymer 28:171-175.
[3] Mason W.R. 2009. Starch use in Foods. In: BeMiller J. & Whistler R. (Eds.). Starch: Chemistry ans
Technology. 3rd edition. Academic Press, Burlington, MA 01803, USA.

1914
 Experimental studies and interpretation of pistachio nut roasting process

Gilles TRYSTRAMa, Reza YEGANEH,b

a
AgroParisTech, INRA, Food process Engineering, F91300 France gilles.trystram@agroparistech.fr
b
Department of Farm Machinery, Faculty of Agricultural Engineering, Ilam University P.O. Box 69315-516, Ilam,
Iran

INTRODUCTION
The pistachio is mainly produced in Iran, USA, and Turkey. The pistachio nut is the second non-
petroleum export product, which has the important role in the development of national economic value
and agro-food industry of Iran. Based on FAO statistics (2005), Iran produced about 275,000 Mt of
pistachio in 2003, which was approximately 54.7% of the world’s pistachio production. Iran exported
184,946 Mt of its pistachio nut in this year and the total export revenue from pistachio was about
679,940,000 US$ in 2003 (Razavi & Taghizadeh, 2007). Roasting is one of the most important processes
giving necessary alterations to the product (Demir & Cronin, 2005). Physical properties also affect on
hydrodynamic / pneumatic conveying characteristics of solid materials, and cooling and heating loads of
food materials (Mohsenin, 1978). Density, shrinkage and porosity are important transport properties that
are widely used in process design calculations, i.e. various heat and mass transfer operations. These
properties are also used to characterize the texture and quality of dry and intermediate moisture foods
(Schubert, 1987). Density is the variable affecting most thermophysical and transport properties.
Shrinkage and porosity are also important parameters in the prediction of diffusional properties of cellular
food during drying (Rotstein, 1987). The objective of the work is to establish a data base of roasting and
its consequences on nuts qualities and to propose an interpretation of the key process variables that
influence product attributes.

RESULTS & DISCUSSION

During pistachio roasting, temperature recording shows the evolution of the internal nuts temperature
during the roasting. It put in evidence the classical effect of air temperature on the evolution of nuts
temperature. Initial nuts temperature increase at all air temperatures during the first 6 minutes of roasting.
Then, the thermal equilibrium is reached and nuts temperature stays constant close to 100 °C where the
balance between evaporation and heat transfers is reached. The duration of the plate is approximately 10
minutes. When constant evaporation period is finished, internal nuts temperature started again to increase,
completing water evaporation, until eventually reaching to value of 104, 123, 135, 144 and 156 °C after
31 minutes for 120, 130, 140, 150 and 160 °C air temperature respectively. At the low temperatures (110
and 120 °C in this study), the status of four zone doesn’t occur during the maximum 31 minutes of
roasting process. It could be seen on curves, that in some cases, the temperature after 6 minutes reaches
more than 100 °C and decrease later. It is probably due to lag in the establishment of the equilibrium.
The relationship between the moisture content and roasting time for pistachio kernels at all temperatures
present a non-linear decrease of moisture content with roasting time. Initially, moisture decreased rapidly
and then the decrease in moisture slowed down considerably as expected. The roasting time performed to
recover the final moisture content varies with the roasting temperature. The experimental data regarding
pistachio nuts moisture variations during roasting at 110-160 °C showed that the time needed to reach
about 15% moisture content was about 30 min for pistachio roasted at 110 °C and about 7 min for
pistachio roasted at 160 °C. The time required to bring the moisture content less as 6% (d.b.) was 23, 18,
14 and 11 min for 130, 140, 150 and 160 °C, respectively. It was observed that the drying rate was higher
at higher temperature, as normally expected.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1915
The water activity of the pistachio kernels was measured at different moisture content and it was observed
a positive correlation and varied from 0.14, 0.15, 0.19, 0.35, 0.60 and 0.64 to 0.97 at 0.332, 0.465, 1.346,
3.242, 10.662, 15.265 and 49.95% (d.b) moisture content respectively. The curves are classical ones,
characterised with a significant decrease in the range of aw: 1 to 0.70 and slower reduction when moisture
content becomes low.
The magnitude of the increase in porosity may be attributed to the change in absolute and apparent
density with the increase in roasting time. The porosity of pistachio kernels increased from 0.159 for raw
pistachio kernels to 0.240, 0.260, 0.267, 0.270, 0.288 and 0.317 at 110, 120, 130, 140, 150 and 160 °C,
respectively. After 10 minutes of roasting, the porosity, for a given temperature does not have significant
evolution. The accuracy of the measurements is not a sufficient to discriminate clearly between curves
due to the determination method. At lower temperature (110 and 120 °C) it seems that the porosity still
have evolution. In numerous applications, when drying occurs combined with heat processing of food, the
creation of porosity is observed, strongly related with moisture departure.
As higher is the moisture content (from 0.332, 0.465, 1.346, 3.242, 10.662 and 15.265% (d.b.)) as lower
is the maximum breaking force: 3312.5, 3075.5, 3190, 3075, 2382.5 and 1945 N to 1355, 1105, 1240,
1085, 1250 and 1350 N respectively and then for raw pistachio increase from there to 1970 N
independently of the temperature. Results show the breaking force is mainly correlated with the moisture
content. Nevertheless, after a minimum value of the breaking force obtain of approximately 30% moisture
content, the force increase. The resulting variation of the force versus moisture content is close to an
exponential variation. The duration of the performed experiment does not permit to observe if there are
any plates for the trends.
An analysis was done to establish the effect of the parameters related with roasting (air temperature and
roasting time) on the properties of pistachio nuts at only 6% moisture content. The results establish
clearly that the time-temperature control of the roasting operation is possible and efficient in order to
obtain a given objective of the physical properties. With increasing roasting temperature, the roasting
time decreased. Aw values of all applications were found under the critical value for long-term storage to
prevent mold growth and aflatoxin contamination, which is at 0.70. Different properties could be obtained
using time temperature operating conditions, and the ranges significant for most of the properties.
This study reported some physical properties of pistachio nuts evolution (Ahmad Aghaei variety)
including nuts temperature, moisture content, apparent volume, apparent density, absolute density,
porosity, water activity and texture. On the basis of our results, it is possible to establish that the moisture
content, roasting temperature and roasting time affect the physical properties of pistachio nuts during
roasting and the following conclusions can be drawn: The curves of nuts temperature presented four
zones: an increase in the first 6 minutes, a temperature constant approximately 10 minutes, an increase
until final the evaporation period and a temperature plateau nearly equal to the air temperature. In the all
temperature, moisture content decrease with increase in roasting time. As the roasting time increased,
apparent volume of pistachio kernels increased at different temperatures studied. The apparent density of
pistachio kernels decreased with increase in roasting time. Initially, in the first 5 minutes, the absolute
density increased and then decrease as roasting time increased. For all temperature, the porosity of
pistachio kernels increased as moisture content decrease. As the roasting time increased, water activity
decrease for all temperature. It is interesting to note that the roasting process could influence the textural
properties of the pistachio nuts, rather than just the influence on the moisture content. The maximum
force of pistachio kernels was found to decrease in the first 5 minutes of process and then increase as
roasting time increased. Other ways are possible to conduct experiment in order to increase heat fluxes to
obtain the new values of physical properties. Roasting using other processing ways could be of interest
(superheated steam, frying for example).

1916
Heat transfer analysis-based prediction of protein denaturation and umami component of

meat during cooking

Naomi Ishiatari, Mika Fukuoka, Naoko Hamada, Noboru Sakai

Department of Food Science and Technology, Tokyo University of Marine Science and Technology,
4-5-7 Konan, Minatoku, Tokyo, Japan (fukuoka@kaiyodai.ac.jp)

INTRODUCTION
In meat cooking, various reactions may occur simultaneously with heat and mass transfer. The
physical properties and eating quality of cooked meat are strongly affected by both the the
degree of protein denaturation and the amounts of umami components resulting from the
difference temperature treatments. Inosinic acid (IMP) is famous as the major component of
umami taste. It seems that the IMP content of meat is affected by the heating temperature
during cooking, because IMP is decomposed by the enzyme originally present in the meat.
However, there have been no reports predicting the changes in protein denaturation and umami
component in accordance with heat transfer during meat cooking. The objective of this work
was to simulate changes in both protein denaturation and the amount of IMP by a 3D finite-
element method. We targeted the vacuum-pack cooking (sous-vide) that is frequently used by
professional chefs.

MATERIALS & METHODS

Beef from the purchase was used for all the experiments. Roast beef was cooked in the
laboratory according to the sous-vide method. We corrected the temperature history for both
the core and surface of the meat and measured the weight and size of the meat before and after
cooking. The kinetic parameters of protein denaturation were measured by differential
scanning calorimetry (DSC). IMP in the meat was quantified by high performance liquid
chromatography (HPLC). The temperature dependency of the IMP decomposition reaction was
also examined in an isothermal heating experiment.

RESULTS & DISCUSSION

Roast beef cooked in the laboratory showed the character with a low weight loss compared
with a usual cooking method. The sample thermogram shows two endothermic peaks. From
published literature, the first peak corresponds to myosin and the second one to the actin of the
fibrillary protein. It turned out the myosin denatures at about 50Υ while the actin denatures at
about 70Υ.The amount of remaining of IMP has decreased as the heating time progresses at
all the heating temperatures. However, as for the sample heated at the temperature above 40Υ,
the amount of remaining was lower than sample heated at 40Υ, because the enzyme activity
that decomposed IMP decreased. Then, the activation energy and the frequency factor of the
IMP decomposition reaction and the enzyme activity decrease reaction were calculated by
using the Arrhenius plot. The shape model similar to the sample cooked in the laboratory was
made, and the three-dimensional heat conduction analysis by the finite element method was

11th International Congress on Engineering and Food - Athens, Greece, 2011 1917
done. Figure 1 shows the comparison between measured value and the calculated temperature
history of meat at the core. The change in the temperature was similar. Next, the protein
denaturation ratio and IMP remaining ratio were simulated by using the protein heat
denaturation rate constant and rate constant related to IMP based on the calculated temperature
history. In case of the sous-vide cooking, it turned out that the denaturation ratio of the actin
was low at the central position and IMP decreased near the central position.

㻝㻜㻜

㼟㼡㼞㼒㼍㼏㼑
㻤㻜 㼏㼛㼞㼑㻔㼑㼤㼜㻚㻕
㼀㼑㼙㼜㼑㼞㼍㼠㼡㼞㼑㻌㻔䉝㻕

㼏㼛㼞㼑㻔㼏㼍㼘㻚㻕
㻢㻜

㻠㻜

㻞㻜

㻜
㻜 㻡㻜 㻝㻜㻜 㻝㻡㻜 㻞㻜㻜
㼀㼕㼙㼑㻌㻔㼙㼕㼚㻕
Figure 1. Comparison between measured and calculated temperature history of meat at the core during
sous-vide cooking.

CONCLUSION
The distribution of thermal denaturation of protein and IMP remaining ratio were analyzed by
heat transfer analysis that imitates sous-vide method. It turned out that the denaturation ratio of
the actin in the central position is low and IMP remaining ratio on the surface is high.

REFERENCES
[1] Wagner J.R. & Anon M.C. 1985. Denaturation kinetics of myofigrillar proteins in brovine muscle.
Journal of Food Science, 50, 1547-1551.
[2] Bertola N. C., Bevilacqua A. E. & Zaritzky N. E., 1994. Heat treatment effect on texture changes and
thermal denaturation of proteins in beef muscle. Journal of Food Processing Preservation, 18(1), 31-
46.

1918
 Effect of steam cooking of food on mass transfer

Emilie DESCOURS1, Eric FERRET 2,3, Nicolas VALANCE 4, Andrée VOILLEY 1,3, Anne-Marie
SEUVRE 1,5

1
Laboratoire EMMA, Université de Bourgogne, 1 esplanade Erasme, 21000 Dijon France
2
Laboratoire GPMA, Université de Bourgogne, 1 esplanade Erasme, 21000 Dijon France
3
Agrosup Dijon, 26 Boulevard Docteur Petitjean, BP 87999, 21079 Dijon Cedex, France
4
Groupe SEB, Rue La Patenee, 21261 Selongey, France
5
IUT Génie Biologique, Université de Bourgogne, Bd Docteur Petitjean, BP 17867, 21078 Dijon cedex,
France

INTRODUCTION
During the last few years, the number of health problems related to poor nutrition has
increased. Due to globalisation and open markets, people have never had such a vast selection
of food products. Therefore, the right choice of cooking preparation can help in overcoming the
problems mentioned above. The most promising method that preserves nutritional as well as
organoleptic food properties, seems to be steam cooking.
The aim of this work is to study the impact of steam cooking against the loss and/or the
generation of aroma compounds in potatoes, especially those that have a major impact in the
flavour of potatoes. Among these, many aldehydes, including hexanal, octanal, nonanal,
decanal, benzaldehyde but also 2-pentylfuran [1], [2]. Hexanal which is present in large
amounts in raw potatoes and in increasing quantities during cooking, is chosen so as the mass
transfer can be studied [3].

MATERIALS & METHODS

600g of potato cylinders (3cm x 2cm) were steam cooked (100°C) for 20 minutes. This amount
of time was determined after studying at three parameters: cooking value, textural measures
and sensorial analysis [4]. After cooking, the water vapour, the potato samples and the
collected cooking juice were analysed.
Quantitative analysis of aroma compounds was done using Chrompack CP9000 gas liquid
chromatograph teamed with a with a flame ionization detector (200°C) (Flow N2:20ml/min,
H2.:26ml/min, air: 360ml/min) The injector temperature was set at 190°C, the column
(stainless steel column 3m length, 2.2 mm inner diameter, packed with Chromosorb 20W/AW,
80/100mesh Carbowax 20M/10%) from 90 to 110°C.
To allow the aroma to be analysed in saturated water vapour, an adapted system was
developed. In a transfer line, the steam was cooled and due to the Peltier effect, the water was
removed and the condensate recovered. After, compounds were concentrated by a thermo-
desorption system (Markes) connected to a Gas liquid chromatograph (7890 A Agilent)
coupled with a mass spectrometer equipped with a quadripole (5975 C Agilent).

RESULTS & DISCUSSION

During cooking, a potato lost on average 0.3% of its initial mass. The water lost represented
48% of water initially introduced in the system.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1919
The first results show a variation of concentration of aroma compounds. To understand the
behaviour of aroma compounds during cooking, hexanal was used as a tracer. This compound
was injected with known quantities into the center or onto the surface of the cylinder before
being steam cooked. The potato, gaseous phase and cooking juice were analyzed after.

Table 1. Average mass balance obtained after steaming potatoes 600g calibrated for 20 minutes.
Mass of hexanal introduced Mass of hexanal after 20 minutes cooking recup
(mg) (mg) and (%)
Without the addition of hexanal 35,4 0,9 1,2
When injected into the center 62,0 0,5 6,3
300ppm (183mg) 34 % 0,3% 3,4%
When injected into the center 27,7 0,9 8,0
300ppm (183mg) 15% 0,5% 4,4%

An important difference was observed if hexanal was injected into the potato or on the surface.
When hexanal was added into the center, only 38% of it was retained by the potato and more
than 62% of the compound released from the cooking system. When we added hexanal on the
surface, the loss was more significant, as only 15% was still in the system. These losses are
greater than the concentration of hexanal as shown in the literature as it is supposed to increase
during cooking.
A solution of hexanal in water at a know concentration was placed in the system to follow the
loss of aroma compound during the cooking. It was observed that after 4 minutes of cooking,
half was lost and after 20 minutes, 98 % of hexanal had escaped out of the system.

Steam cooking is an open system that works under atmospheric pressure. The compounds
responsible for the flavour are mainly hydrophobic aromatic compounds (log P>1,5) especially
aldehydes that, under these conditions, release from the system.

CONCLUSION
Most aroma compounds having a strong impact on the flavour in steam cooked potatoes are
aldehydes. These hydrophobic compounds are weakly retained by the low-fat foods cooked in
a saturated water vapour atmosphere. The steam cooking will cause a loss of aroma
compounds. It is necessary to improve steam cooking to try to preserve the maximum amount
of compounds to limit lost.

REFERENCES
[1] Shelley H. Jansky. 2010. Potato Flavor. American Journal of Potato Research, 87, 209217.
[2] Mutti, B., and W. Grosch. 1999. Potent odorants of boiled potatoes. Nahrung/Food 43: 302306.
[3] Petersen, M. A.; Poll, L.; Larsen, L. M. 1998. Comparison of volatiles in raw and boiled potatoes
using a mild extraction technique combined with GC odour profilingand GC-MS. Food Chem. 61,
461- 466.
[4] Chiavaro E., Barbanti D., Vittadini E., Massini R. 2006.The effect of different cooking methods on
the instrumental quality of potatoes. Journal of food engineering, 77, 169-178.

1920
Development of experimental devices in order to study the interactions between heat and

mass phenomena and thermal degradation reactions of lipids during domestic reheating

of pre-fried food products

Cernela, J.a,b, Heyd, B.a,b, Broyart, B.a,b

a
AgroParisTech, UMR 1145 GENIAL Food Process Engineering, 1, avenue des Olympiades, 91744
Massy cedex, France (bertrand.broyart@agroparistech.fr)
b
INRA, UMR 1145 GENIAL Food Process Engineering,1, avenue des Olympiades, 91744 Massy cedex,
France

INTRODUCTION
When solid products containing lipids are submitted to high temperatures, thermal degradation reactions
of lipids occur. Few studies concern thermal degradation reactions of lipids during contact heating or
oven cooking of food products. Some experimental and theoretical studies can be found concerning
single- or double-sided pan frying of beef burgers: [1], [2], [3]. With regards to these applicative and
scientific stakes, a French national 3-years research project called DOMINOVE (“DOMestic heating of
deep-fat fried products IN OVEn and pan”) has been supported by the French Research National Agency
(ANR) since 2010. This paper will first present original results taken from an experimental campaign
aiming at evaluating the variability inherent to hot-air cooking and contact heating at domestic scale. In a
second part, the experimental laboratory devices developed in our laboratory to reproduce the domestic
operations of concern and their thermal characterization will be presented and discussed.

MATERIALS & METHODS

For the study of the variability inherent to domestic hot-air cooking, three commercial electric ovens were
used: two built-in ovens and a mini-oven (free or forced convection). After a pre-heating step, an
aluminum cooking plate containing 3 breaded poultry products was put at the centre of the oven cavity.
During these experiments, the product thermal environment was assessed by three variables: the air
temperature, the convective heat transfer coefficient and the equivalent radiative temperature (with heat
fluxes sensors).
For contact heating, four commercial heating devices typically encountered in domestic conditions were
used and two Teflon®-coated pans were selected: a thin and light aluminium pan (pan #1) and a heavier
one made from an assembly of stainless steel and aluminium (pan #2). Pan temperature was measured
using a thermocouple inserted in a thin hole below the pan surface. After a pre-heating step, a thermal
load of 3 breaded poultry products were put in the pan and the selected heating power is adjusted to a
medium or high level. This operation is made with or without sunflower oil. The heterogeneity of the pan
surface temperature was evaluated just before the disposal of the products using an infrared thermography
camera.
For hot-air cooking and contact-heating devices, the electrical power consumptions were recorded during
all experiments with a Joule-meter.

RESULTS & DISCUSSION

Characterization of domestic heating devices: the fastest pre-heating is achieved with the induction
hob. The addition of an oil layer has no significant effect on pre-heating rate except for induction hob. By

11th International Congress on Engineering and Food - Athens, Greece, 2011 1921
contrast, the nature of the pan has an effect on the pre-heating rate. The heterogeneity of pan surface
temperature at the end of pre-heating depends widely on the type of pan selected and the nature of the
heating device.
Once food products have been put on the pan, the four contact-heating devices show very different
behaviours. Pan temperature is never constant during heating. Trials at high power show a pan
temperature always rising and reaching circa 300°C.
When looking at the pre-heating rate for the hot-air cooking devices, it can be noted a good accordance
for the two built-in ovens tested. In the mini-oven the pre-heating rate is significantly higher although the
power consumption is lower.
Very different air temperature evolutions during cooking are also observed for hot-air cooking devices.
This is mainly due to the different modes of temperature control within the oven. Whatever the oven, the
set point is never successfully reached and kept constant during cooking. In the three ovens selected, the
convective heat transfer coefficients measured at the centre of the cooking plate in free convection are
nearly the same. In contrast for the forced convection mode, we note a significant difference between the
two built-in ovens and the mini-oven. The results on equivalent radiative temperature are more difficult to
interpret because of the unstable air temperature.
Development of laboratory heating devices: the experimental device for hot air cooking was developed
from a commercial oven. The air temperature control of the commercial oven was improved by adding a
PID controller to the installed electronic control system. The convection mode has an influence on the
value and heterogeneity of the convective heat transfer coefficient and equivalent radiative temperature
over the cooking plate.
Contrary to the hot air cooking device, the laboratory device for contact heating is an original prototype.
A stainless-steel pan with a bottom made of a sandwich stainless steel/aluminium/stainless steel is
positioned on a double jacketed stainless-steel cylindrical basis filled with an insulating material. A
heating resistance is fixed inside the basis. A thermocouple inserted in the centre of the pan below the
surface is used for controlling the pan temperature using a PID controller. The two main advantages of
this device are the good surface temperature uniformity of the pan and the high thermal inertia of the
system coupled to the pan temperature control which allows to work in constant and repeatable
conditions.

CONCLUSION
During domestic hot-air or contact cooking, the behaviour of the consumer and the nature of equipment
used influence greatly the thermal environment of the heated product and hence certainly the magnitude
of heat and mass transfers and reactivity of lipids within the product. These observations led us to develop
two heating devices in order to study heat and mass transfers in repeatable and constant conditions: a hot-
air heating device based on a commercial oven and an original prototype for contact-heating.

REFERENCES
[1] Ikediala, J.N., Correia, L.R., Fenton, G.A., Ben-Abdallah, N. (1996). Finite-element modelling of heat transfer in
meat patties during single-sided pan-frying. Journal of Food Science, 61, 796-802.
[2] Pan, Z., Singh, R.P., Rumsey, T.R. (2000). Predictive modeling of contact-heating process for cooking a
hamburger patty. Journal of Food Engineering, 46, 9-19.
[3] Ou, D. & Mittal, G.S. 2006. Double-sided pan frying of unfrozen/frozen hamburgers for microbial safety using
modelling and simulation. Food Research International, 39, 133-144.

1922
The Effect of UHT and VAT Thermal Processing Systems on Whey Protein Denaturation

and Gel Strength of yoghurt

Labropoulos, A a., Varzakas, T., b, Anestis, S.a

a
TechnologicalEducational Institute of Athens, Hellas (athanlab@teiath.gr)
b
Technological Educational Institute of Kalamata, Hellas (tvarzakas@teikal.gr)

INTRODUCTION
It is well known that different process variables can induce changes in the physical properties of milk
proteins. Such variables affecting physical properties of proteins include heat treatment, protein content,
acidity, etc. There is considerable controversy, however, regarding temperature-time relationships
necessary to change the physical property of a product, e.g. stability. Therefore, it is of interest to
determine the degree of whey protein denaturation induced by different processing systems and the
correlation of denaturation to the gel strength. The current study examines the effect of two process
systems i.e. continuous UHT and batch VAT heat treatments, on the denaturation of whey proteins with a
view to further correlation with the mechanism of gel strength in yoghurt.

MATERIALS & METHODS

Raw whole milk was heat treated by two processing systems, i.e. a continuous UHT and a batch VAT,
represented by the following temperature-process holding time combinations: UHT processing at
temperatures 132 0C to 154 0C with an app. 6 0C increment for 0.0, 1.2, 3.3, 5.2, 9.0, and 12.0 sec and
VAT processing at temperatures 66 0C to 88 0C with an app. 6 0C increment for 0.0, 5.0, 10.o, 20.0, and
30 min. Unheated milk was utilized as a control.
UHT and VAT heat treatments were carried out in a helically coiled tube, with an indirect heating system,
automatic temperature control and in a steam jacketed batch type pasteurizer respectively. Different
residence times were obtained by using holding tubes of varying length size and changing pump speeds in
the UHT process system. All heat treatments were homogenized at 60 0C preheating temperature and an
approximate 105 Kg/cm2 operating pressure. ). A Cherry-Burrel curd tension meter was used to measure
the strength of yoghurt gels. Whey proteins were separated on cellulose acetate strips using an
electrophoresis procedure. Qualitative analysis of the whey protein fractions was made according to the
procedure of Puyol et al. in which the whey protein fractions of the milk are identified by the rate of
travel in the electric field relative to standard proteins.

RESULTS & DISCUSSION

Effect of heat treatment on whey protein electrophoresis
A typical densitometer tracing of separation of whey proteins of raw milk shows 5 distinct electrophoretic
peaks, corresponding to immunoglobulins (Igs), alpha-lactalbum in A (alpha-La), beta-lactoglobulin B
(beta-Lg B), beta-lactoglobulin A (beta-Lg A), and bovine serum albumin (BSA).
Progressive decreases in the area of electrophoretic peaks occurred with an increase in process holding
time, for example, at the UHT system (149 0C) and at the VAT system (82 0C) (Figure 1). Qualitative
analysis of the areas under the electrophoretic peaks for the UHT treatments at 1320C through 154 0C
showed that these treatments with no process holding times did not severely denature the whey proteins.
A significant denaturation was observed at l49 0C and above. However, the same UHT treatments but

11th International Congress on Engineering and Food - Athens, Greece, 2011 1923
with 12 sec process holding times yielded a complete denaturation for all the temperatures applied to
whey proteins.
On the other hand, VAT treatments at 66 0C from 0 to 30 min process holding times indicated only a
small effect on whey protein denaturation. There was a severe denaturation effect somewhere between 82
0
C and 88 0C temperatures. When VAT heat treatments were applied at various temperatures with a
constant 30 min process holding time, a substantial denaturation effect was obtained after 71 0C
temperature.
Effect of heat treatment on gel strength
Whey proteins have been demonstrated to participate in gel formation, while other studies have shown
that heat treatment (conventional or UHT) of milk affects the rheological properties of yoghurt gel
prepared from this milk. However, a differential effect of VAT and UHT heat treatments on the gel
strength of yoghurt made by the above processing systems was observed in this study. Gel strength
measured by curd tension values ranged between 15 and 34.5 g for the UHT heat treatments and reached
up to 85 g for the VAT heat treatments (Table 1). The strength of yoghurt gels prepared from UHT heat
treatments was, thus, always lower than those prepared from VAT heat treatments

Table 1. Effect of UHT and VAT heat treatments on gel strength of yoghurt
HEAT TREATMENT Process Holding Gel
Process Temperature
Time 0
Strength
( C)
(sec) (g)
UHT 0.0 149 24
UHT 3.3 149 33
UHT 5.2 149 27
UHT 9.0 149 14
UHT 12.0 149 12
VAT 1800.0 82 86

CONCLUSION
The results of this study indicate that electrophoresis on
cellulose acetate membranes provides a relatively simple
and quick procedure for separating and measuring the
relative quantities of protein denaturation induced by
different thermal processing operations. Denaturation of
the whey proteins induced by UHT and VAT processing
systems was dependent on thermal processing systems and
thermal processing parameters (process holding time and
temperatures). A rough correlation between extent of whey
protein denaturation and gel strength was found for the
VAT heat treated whey proteins, while no such correlation
was observed in the UHT heat treated whey proteins. On
the other hand, the strength of gels prepared with the UHT
Figure 1: Electrophoretic patterns of whey
process was always lower than those prepared with the
proteins of whole milk thermally processed
VAT process. This could lead to production of light
by the UHT system at 149°C temperature
strength gels with an application to drinkable or semi-
and 0 to 12 sec process holding times
liquid products.

1924
 Application of ohmic heating to whole egg

Toshio Nakai, Mika Fukuoka, Noboru Sakai

Department of Food Science and Technology, Tokyo University of Marine Science and Technology,
4-5-7 Konan, Minatoku, Tokyo, Japan (fukuoka@kaiyodai.ac.jp)

INTRODUCTION
Ohmic heating has various advantages such as the high heating efficiency and the accuracy of
the temperature control, and it is applied to the sterilization of liquid food in the foods industry.
Because the liquid egg is delicate liquid food changed from the liquid phase to a semisolid
state so that the protein may denature according to the heating temperature, the heating
sterilization is very difficult. Objective of this study is to examine whether the ohmic heating
can be applied to the thermal processing of the whole egg by understanding the electrical
property of the egg component.

MATERIALS & METHODS

Whole egg: Fresh shell eggs (large size, 52-76g) within one week from the date of spawning
purchased from a supermarket were used for heating experiments and impedance
measurements
Ohmic heating system: Ohmic heating system was composed of the power supply unit
(frequency at 20kHz, 3kW in capacity, applied voltage range 0~100V), the heating container
with a titanium electrode (L53mm×W 79mm×T1mm), PC for data storage.
Impedance measurement: The whole egg was separated to egg white and the whole yolk with
the separator, egg white was put into the SUS basket of 1mm mesh, the egg white that
remained in the basket was assumed the dense egg white and egg white that had passed
naturally were assumed to be the liquid egg white. Yolk w/ and w/o vitelline membrane were
used respectively. Liquid whole egg was prepared by stirring whole egg without the chalazae.
The impedance of those samples was measured by LCR meter (LCR HIGHTESTER 3532-50,
HIOKI Co., Japan).

RESULTS & DISCUSSION

A part of egg white became clouded by ohmic heating at 50V and the protein denaturation was
confirmed. The region which became clouded was a dense egg white that exists around the
yolk. It was clarified that the thermal denaturation of the dense egg white in the vicinity of the
center between two electrodes was remarkable. The temperature histories of both yolk and egg
white during heating were shown in Figure1. Egg white was heated selectively during the
ohmic heating (50V-2min) of the whole egg. On the other hand, the temperature rise at the
whole egg was small though the whole yolk enclosed by the dense egg white and the liquid egg
white was the central part between two electrodes.
From the result of the heating experiment, the impedance measurements for component of the
egg were necessary. Figure 2 shows that the temperature dependence of the impedance of the
egg components. The frequency of impedance measurement was 20 kHz.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1925
 Figure 1. Temperature history of whole egg during ohmic heating at 50 V.

The impedance of egg white and yolk has become small with the temperature rise and the
temperature dependency of impedance was able to be confirmed. On the other hand, the
impedance of both dense egg white and liquid egg white were small. Moreover, there is little
temperature dependence in egg white. Impedance at 50 kHz was also measured. The
impedance of whole yolk has increased remarkably compared with that of 20 kHz, though that
of yolk and egg white didn't change.
It was thought that a big difference in the electrical property between egg white and yolk
affected the characteristic in the ohmic heating of the whole egg.

250
wholeyolk

yolk
200 denseeggwhite

liquideggwhite

) 150 liquidwholeegg
(O
ce
n
a
d
e
p
i 100
m

50

0
20 30 40 50 60 70 80
temperature(Υ)

Figure 2. Temperature dependency of the impedance at 20 kHz of frequency.

1926
 Transient mass and heat transfer during potato deep fat frying – The effect of the oil

type, frying load and initial frying temperature

John S. Lioumbasa, Margaritis Kostogloua, Thodoris D. Karapantsios

a
Division of Chemical Technology, Department of Chemistry, Aristotle University of Thessaloniki,
Thessaloniki, Greece (karapantsios@chem.auth.gr)

INTRODUCTION
In order to design frying equipment for space, the critical points of the frying process under no
or partial gravity must be examined. On this account, ESA (European Space Agency) has
initiated a project to examine the effect of different g-levels on frying. Experiments at
increased gravity levels are conducted in ESA‘s Large Diameter Centrifuge whereas
experiments at reduced gravity levels are conducted during ESA’s Parabolic Flight Campaigns.
Beforehand, meticulous experiments are performed under normal (terrestrial) gravity
conditions to serve as reference. This is indeed the objective of this work.
During long-duration space missions astronauts must receive food that is not only nutritional
but should also offer pleasure which is significant for crew morale and performance. Frying is
a popular culinary process worldwide. This is especially so regarding potato frying in western
societies. So, from a psychological point of view it would be important for astronauts to
include fried potatoes in their menu. Frying is a process strongly influenced by the acceleration
of gravity. It is not only the convective heat transfer between the oil and the food surface that
is affected by gravity. What is perhaps more significant is the formation and removal of
evaporated water in form of bubbles from the food surface that leaves behind a porous crust
which dictates the eventual oil uptake and food oral (sensory) perception.
To our knowledge, Hubbard & Farkas [1, 2] were the first and only researchers, who
performed simultaneous on-line weight loss measurements and temperature measurements
inside the potato, during deep fat frying (large scale frying) of single potato pieces. Hubbard
and Farkas [1] focused on frying of a single potato piece (or just a few pieces) assuming
constant thermophysical properties of both the oil and the food. However, this does not
resemble the conditions encountered in actual applications where many food items fry together
in close proximity resulting to an oil bath temperature drop of 30 to 45 oC. The results of the
present study, apart from acting as reference for future experiments under increased and
reduced gravity levels, improve our understanding of the complicated phenomena prevailing
during large scale frying and so contribute to the design of more energy-efficient frying
appliances.

MATERIALS & METHODS

This work investigates the dependence of transient mass transfer characteristics (i.e. vapour
flux) and heat transfer characteristics (i.e. heat transfer coefficient and thermal conductivity of
the fried medium) during potato deep frying, on the type of frying oil, the initial frying
temperature and the frying load. Simultaneous on-line water loss measurements as well as
temperature recordings both inside the potatoes (Angria variety, all from the same producer,
geographical region and harvesting period) and in the oil are employed to evaluate mass and

11th International Congress on Engineering and Food - Athens, Greece, 2011 1927
heat transfer characteristics. Two types of oil (i.e. extra virgin olive oil and refined palm oil),
two initial frying temperatures (i.e. 150 and 180oC) and two frying loads (i.e. 1/35 and 1/7
kgpotatoes/Loil) are examined. These two potato-to-oil ratios are typical for large scale catering
and industrial applications, respectively.

RESULTS & DISCUSSION

The oil temperature, Toil, profiles and the water loss, mw, data per potato stick are presented in
Figure 1 and 2 respectively (for initial Toil =180 oC and two frying loads).

200 2.0
Initial oil temperature: 180o C initial oil temperature: 180oC (b)

Frying load: 1/35

180
1.5

mw per stick, gr
160
Frying load: 1/35
Toil , o C

1.0
Frying load: 1/7
140

Frying load: 1/7

0.5
120
olive oil olive oil
palm oil (b) palm oil
100 0.0
0 50 100 150 200 250 300 0 50 100 150 200 250 300
t, sec
t, sec

Figure 1. Oil temperature profiles during Figure 2. Average loss of water mass per
frying at two frying loads at initial oil potato stick during frying at two frying
temperature 180oC. loads at initial oil temperature 180oC.

CONCLUSION
It is found that the frying load and the initial frying temperature significantly affect the Toil
profiles (Figure 1) as well as the mass of the evaporated water (Figure 2). On the contrary,
virtually no significant dependence on the oil type is observed. Furthermore, by employing a
fundamental mass and heat transfer analysis, different zones have been identified within the
potato exhibiting distinct properties.

ACKNOWLEDGEMENTS: This study was carried under the programme “Influence of

gravity conditions on mass and heat transfer in porous media” and funded by (Co. No.
22470/09/NL/CBi). The view expressed herein can in no way be taken to reflect the official
opinion of the European Space Agency.

REFERENCES
[1] Hubbard, L.J. and Farkas, B.E., 1999. A method for determining the convective heat transfer
coefficient during immersion frying, Journal of Food Process Engineering, 22, 201-214.
[2] Hubbard, L.J. and Farkas, B.E., 1999. Influence of oil temperature on convective heat transfer during
immersion frying. Journal of Food Processing Preservation 24, 143–161.

1928
 Acceptance of Iron Fortified Rice (I-Rice) in the Philippines to Combat Iron Deficiency

Anemia (IDA)

Edith M. San Juan, Neri O. Camitan, Amelita C. Natividad, Mario U. Gochangco, Lauro D. Alkuino,
Alberto R. Cariso, Jr., Dr. Alicia O. Lustre and Dr. Amelia W. Tejada

National Food Authority-Food Development Center, Taguig City, Philippines (info@fdc.net.ph)

INTRODUCTION
Rice is a valuable vehicle for alleviating iron micronutrient deficiency because of its role as the
basic staple food for 93% of households in the Philippines [1]. For rice to be an effective
delivery vehicle for iron, its color, flavor and price should be similar to that of good quality
non-fortified rice. It has to be white in color, with the flavor of good quality rice and at a price
similar to the latter. These characteristics are not easy to achieve in I-Rice due to the effect of
adding iron on the color of the cooked rice, and on its price. Both color and price are factors
that affect the purchase intent of rice of low income consumers.

The main objective of the study was to assess consumer acceptance of I-Rice to household
members of Grade 1 students from five identified schools in the Philippines who were either
participants or not participants in the Food for School Program (FSP) of the Department of
Education (DepEd) [2]. The specific objectives were: (a) to identify the sensory attributes
preferred by Filipinos in I-Rice; (b) to determine which of the demographic profiles of the
respondents affect the acceptance of I-Rice; and (c) to establish the relationship of the different
socio-demographic profiles of the respondents and acceptance of I-Rice.

MATERIALS & METHODS

The surveys were conducted in five (5) public elementary schools, namely, Tenement
Elementary School (TES) in Taguig City, Camarin D Elementary School (CDES) in Caloocan
City, Soledad Marasigan Elementary School (SMES) in Pili, Camarines Sur, North City
Elementary School (NCES) in Dumaguete City, Negro Oriental, and Libertad Central
Elementary School (LCES) in Butuan City, Agusan del Norte using the home-use test (HUT)
protocol. Of the five schools, TES and CDES are schools participating in the FSP while the
other three schools are not participating. The five schools were considered to determine if there
is significant difference in responses between the two classifications of schools. The target
respondents were the qualified household members of Grade 1 students. A total of 2,442
respondents participated in the study. The materials used for the survey were as follows: (1)
Letter to the Households; (2) Instruction Sheets; (3) Consent Form; (4) Demographic
Questionnaire – Parts 1 and 2; (5) Scoresheet; and (6) I-Rice samples. The 9-point hedonic
scale was used to determine the acceptance rating for odor, color, flavor, and overall liking of
I-Rice.

RESULTS & DISCUSSION

Results of non-parametric test using the Kruskal-Wallis Test for group comparison revealed
that there were no significant differences among the acceptance ratings obtained from the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1929
respondents of the five (5) schools as shown in Table 1. This means that the responses of all
the qualified respondents on acceptance of I-Rice were the same in all schools regardless of
whether the chosen school is participating or not in the FSP of DepEd.

Table 1. Kruskal-Wallis Test of significance for acceptanceof I-Rice in five (5) schools
Test Color Odor Flavor Overall
Chi-square 7.383 245.291 6.403 0.801
d.f. 4 4 4 4
Asymp. Sig. 0.1170 0.0000 0.1710 0.9272
*Significant at 0.05 level.

Statistical analysis using Spearman correlation showed that demographic profiles of the
respondents commonly portray a weak positive correlation on acceptance of I-Rice as indicated
by the correlation coefficient of age, frequency of eating rice, amount of rice consumption, and
willingness to buy I-Rice. A weak negative correlation was also observed between educational
attainment and acceptance of I-Rice. The rest of the demographic profiles that were statistically
analyzed using the Spearman correlation (awareness of FSP of the government, experience of
eating I-Rice, awareness of health benefits of I-Rice, and price willing to pay for I-Rice)
showed a combination of weak positive and weak negative correlations on acceptance.
Gender of the respondents showed to have a significant difference on acceptance for overall
liking of I-Rice using Mann-Whitney (Wilcoxon). Other demographic profiles such as marital
status, number of parents living with Grade I students, job nature of the head of the family, and
employment status of the income contributor of the household were found to have no
significant difference on acceptance of I-Rice, except for odor in relation to job nature of the
head of the family and employment status of income contributor. This means that only the odor
attribute gained high consideration on acceptance of I-Rice from the preference of the
respondents.

CONCLUSION
Results of non-parametric test revealed that there were no significant differences among the
acceptance ratings obtained from the respondents of the five (5) schools. Statistical analysis
using Spearman correlation showed that among the demographic profile of the respondents,
age, educational attainment, amount of rice consumption and their willingness to buy I-Rice
had significant effects on its acceptance. Gender of the respondents showed to have significant
difference with each other with respect to overall liking of I-Rice using Mann-Whitney
(Wilcoxon) Test. The findings imply that moderate acceptance of I-Rice could be influenced
by socio-demographic profiles of the consumers that were found to have slight relationship on
acceptance.

REFERENCES
[1] Food and Nutrition Research Institute (FNRI). 2003. Presentation Power point provided during
interview on 14 June 2007. Manila, Philippines.
[2] Food for School Program (FSP) Inter-agency Technical Working Group. 2006 Operational
Guidelines on the Food for School Program “Bigas para sa Mag-aaral at Pamilya. February 2006. A
joint project of the National Food Authority, Department of Agriculture, National Nutrition Council,
Department of Education and Department of Health. pp. 13.

1930
 Quality characteristics and drying behaviour of muffins baked in steam assisted and

convectional ovens

Sakin Yilmazer, Ma., Isleroglu, Ha., Kemerli, Ta., Ozdestan, Oa., Guven, Gb., Kaymak-Ertekin, Fa.,
Uren, Aa., Ozyurt, Bc.

a
Ege University, Faculty of Engineering, Izmir, Turkey (melike.sakin@ege.edu.tr,
hilal.isleroglu@ege.edu.tr, tanselkemerli@gmail.com, ozgul.ozdestan@ege.edu.tr,
figen.ertekin@ege.edu.tr, ali.uren@ege.edu.tr)
b
Ministry of Agricultural and Rural Affairs, Izmir Province Control Laboratory, Bornova-Izmir, Turkey
(guvengonul@yahoo.com)
c
Arçelik A.S. ÇayÕrova Campus, ARGE Directory,
Material Technology Department, Istanbul/ Turkey (bekir.ozyurt@arcelik.com)

Advantages of steam-baking and natural and/or forced convection baking are shared in steam
assisted baking which attracts increasing interest with the increasing health standards of the
society. Steam assisted baking (steam+forced convection/turbo) is known as resulting in
healthy foods.

Baking muffin by using steam assisted oven is not common as it is with bread. By the effect of
steam assisted baking of muffins, the idea of lowering the formation of harmful chemical
compounds, such as acrylamide, and keeping the physical quality at the same time was driven.
Muffins by steam assisted baking were produced and it was evaluated the quality
characteristics as a profile, as well as the drying behaviour during a complete baking process.

Steam assisted baking oven used in the study is a hybrid oven having an inner steam generator.
The steam generated from 150-200 g water was injected into the oven cavity in three times
during the baking process. The steam injection times were determined by preliminary
experiments. The average moisture content, temperature profiles at the top surface, inner
position of muffin and oven ambient and quality characteristics (height, bulk density, surface
colour (Hunter a and browning index (BI) value) and acrylamide content) of muffins baked at
steam assisted oven and natural and forced convection ovens at 160 qC were determined.
Drying behaviour was observed making use of the total moisture loss values.

Steam assisted oven was not resulted in a significant difference in average moisture content of
muffins, compared to natural and forced convection ovens (p>0.05), which is an important
criteria in packaging and storage stage. Temperature profiles obtained at different ovens do
nearly match. The oven ambient temperatures during steam assisted baking was a little lower
than the other ones although the set values were all constant, due to the effect of condensation
of steam injected to the oven chamber. Surface Hunter a and browning index (BI) values of the
muffins were observed that steam assisted oven had significantly lower values than the natural
and forced convection ovens at all baking times (p<0.05). The acrylamide content was
determined to be at a lower (22.56 (±0.16) ppb) value for steam assisted baked muffins, where

11th International Congress on Engineering and Food - Athens, Greece, 2011 1931
it was 62.73 (±4.68) ppb for forced convection and 59.5 (±13.12) ppb for natural convection
baked muffins, each for 40 minutes baking at 160qC. The drying rates calculated for the three
ovens were compared and the maximum drying rate was observed for the forced convection
oven baked muffins, where the steam assisted ones showed the minimum drying rates. This
would mean that it would be required a longer baking time for steam assisted baking to reach a
similar eventual moisture loss value.

It was seen that steam assisted baking would be a good baking choice over natural convection
or forced convection baking in that it was resulted in lower acrylamide content, and Hunter a
and BI values, and nearly same moisture content that would be recommended for better quality
muffins.

1932
 Study Of An Innovative Combination Between Microwaves And Enzymes Applied To

Bakery Products.

T. De Pilli*, A. Derossi, R. Giuliani C. Severini

University of Foggia, Department of Food Science, via Napoli, 25, 71100 Foggia, Italy
(t.depilli@unifg.it)

INTRODUCTION
The biggest difference between convective ovens and microwave ovens is the inability of the
microwave ovens to induce browning. The cool ambient temperature inside a microwave oven
causes surface cooling of microwave-baked products and low surface temperature prevents
Maillard browning reactions from being formed, these are responsible for the production of
many flavoured and coloured compounds.
The aim of this research was to combine enzymatic and microwave treatments in order to
improve sensorial characteristics of products baked by microwaves.

MATERIALS & METHODS

The dough was prepared mixing wheat flour, running water and different amounts of enzymes
like amylase (GRINDAMYL Amylase 1000 from Aspergillus orizae - 1000 FAU/g); protease
(GRINDAMYL Protease 41 from Bacillus subtilis - 41 FAU/g) and glucoamylase
(GRINDAMYL AG 1500/C from Aspergillus orizae - 1500 FAU/g) supplied from Danisco
Cultor (Grindsted, Danimarca). For each power percentage, the treatment times were chosen to
obtain samples with the same value of water activity (0.1-0.3). A factorial design at five
variables (power percentages, time of enzymatic pre-treatments, amounts of amylase,
glucoamylase and protease) and five levels was elaborated by Central Composite Design.
Percentages of reducing sugars developed during enzymatic treatment were measured
according to the method proposed by Tateo [1]. Colour on the surface of samples was
determined from image processing by software Image-Pro Discovery 6.5. Bulk density was
determined according to Bhatnagar et al. [2]. Breaking strength (N/mm2) was measured by a
dynamometer Stable Micro System TA-HDi Texture Analyser (ENCO s.r.l., Venezia, Italy).

RESULTS & DISCUSSION

Results relating to amounts of reducing sugars developed in samples during enzymatic
treatment show a decrease at the highest amounts of amylase and glucoamylase. In particular,
the maximum production of reducing sugars was obtained in dough added of middle amounts
of three enzymes (0.05 g/kg of dough for amylase and glucoamylase and 0.5 g/kg of dough for
protease). An amount of enzyme more then 0.05 g/kg caused, probably, a production of
reducing sugar such as to inhibit enzymatic activity and to change the reaction equilibrium
toward reagents. Moreover, it is known that high amounts of glucoamylase cause an opposite
reaction i.e. these conditions catalyze the repolymerization of glucose into isomaltose. An
interaction between oven power percentages and enzymatic treatment times was observed; it is
possible to obtain samples with best colour characteristics increasing enzymatic treatment
times and oven power percentages. These operating conditions determine a decrease of thermal

11th International Congress on Engineering and Food - Athens, Greece, 2011 1933
treatment time and therefore an energy saving. The increase of protease amount determines an
increase of colour percentages in absence of glucoamylase and middle amounts of amylase,
this could due to the increase of amino acid production that is one of reagent of Maillard
reaction. Nevertheless, the presence at the same time of the highest amounts of protease,
glucoamylase and amylase involved a decrease of colour percentages. This could be attributed,
probably, to inhibition of amylase and glucoamylase by Amadori compound that caused a
decrease of reducing sugar formation [3]. Nevertheless, middle amounts of three enzymes
determined a good characteristics of colour. A considerably decrease of bulk density was
obtained to short enzymatic treatment times with increasing of power percentages because of
increase of spring oven that occurred under these operating processing conditions. With regard
to breaking strength (N/mm2), the only enzyme that showed a significant effect on this index
was protease (Figure 1). In particular, the increase of enzyme amount involved a decrease of
breaking strength values of samples, which resulted more fragile.

Figure 1. Break strength values (N/mm2) of roasted samples as a function of amounts of

protease and enzymatic treatment times.

CONCLUSION
Results showed that the best operating conditions, to obtain a good colour and the lowest bulk
density (0.38 g/ml) of samples, were the highest time of enzymatic pre-treatment (30 minutes)
and power percentages of 90%. All enzymes had a positive effect on colour and texture of
samples only if added together and at middle amounts (0.5 g/kg protease and 0.05 g/kg
amylase and glucoamylase), which corresponds to maximum formation of reducing sugars and
did not involve an inhibition of enzymatic activity. Moreover, the protease was the only
enzyme that showed a positive and significant effect on decrease of breaking strength values.

REFERENCES
[1] Tateo, F. 1969. A spectrophotometric method for the analysis of mixtures of maltose, glucose and
sucrose in foods. Rassegna-Chimica, 21(4), 174-77. [2] Bhatnagar S., Hanna, M.A. & Lincoln, NE. 1997.
Modification of Microstructure of Starch Extruded With Selected Lipids. Starch/Stärke, 49, 12-20. [3]
Woods, L.F.J. & Swinton, S.J. 1991. Enzymes in the starch and sugar industries. In: Tucker G.A. &
Woods L.F.J. (Eds), Enzymes in Food Processing, AVI, Van Nostrand Reinhold, New York.

1934
 Effective removal of heavy metal in some fish sauce products by tannin treatment

T. Sasakia, T. Michihataa, S. Nakamuraa, T. Enomotob, T. Koyanagib,

H. Taniguchib, M. Aburatanic, M Koudoua,b, K. Tokudac

a
Industrial Research Institute of Ishikawa, Kanazawa, Japan (t-sasaki@irii.jp)
b
Ishikawa Prefectural University, Nonoichi, Japan (enomoto@ishikawa-pu.ac.jp)
c
Shata Shuzo Co., Ltd., Nonoichi, Japan (tokuda@tengumai.co.jp)

INTRODUCTION
Fish sauce is a traditional seasoning produced by fermentation of fish or mollusc under high
concentration of sodium chloride for one or two years. It contains quite high amounts of free
amino acids and peptides [1], and furthermore, has been shown to possess health effective
functions such as anti-oxidative and angiotensin I-converting enzyme (ACE) inhibitory
activities. It is reported that a part of fish sauce products contain certain amounts of heavy
metals (i.e., cadmium (Cd), arsenic (As), lead (Pb)) like other seafoods [2]. Heavy metals in
these fish sauces are derived from fish or mollusc organ, which is supposed to intake and
accumulate heavy metal from polluted seawater. The found levels of heavy metals are
considered not to be harmful as the daily intake of these fish sauces is quite low. Still it is
desirable to remove heavy metals from fish sauce products. There have been many reports on
the removal of heavy metals from fish or mollusc organs [3]. In most of these works, strong
acids, such as sulfuric acid, are used to remove heavy metals, but these acids are not acceptable
for processing of foods including fish sauce. The purpose of this research is to develop new,
inexpensive and acceptable method for removing heavy metals from fish sauce. Particularly,
we tried to remove Cd from fish sauce produced from squid. We assumed that Cd molecules
are present in protein-bound forms in fish sauce as well as in fish or mollusc organs. Therefore,
we surveyed food additives that can remove metal-bound proteins from fish sauce by selective
precipitation without influencing on other components and biological functions.

MATERIALS & METHODS

Tannin, glucono delta-lactone (GDL) and bentonite, all approved as food additives in Japan,
were tested as clarification agents. These agents were added respectively to fish sauce, and
after shaking for a few seconds, the mixtures were centrifuged at 8000 X g for 10 min to obtain
treated fish sauces. The Cd concentrations in fish sauce were analyzed by ICP-AES. In
addition, the chemical component (total nitrogen, free amino acids) and biological functions
(1,1-diphyenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and ACE inhibitory
activity) were also evaluated.

RESULTS & DISCUSSION

When the Cd, As, Pb and Hg concentrations were analyzed in various fish sauces, Cd was
detected in some of fish sauce manufactured from squid, and the concentration was higher than
those of other fish sauces. We then tried to remove Cd from the fish sauce by using various
food additives described above. In this experiment, tannin was found to be most effective in
removing Cd.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1935
Figure 1 shows the effect of tannin concentration on the Cd concentrations in treated fish
sauces. The Cd concentrations in the treated fish sauces decreased with increase in the amounts
of tannin added Then the Cd level decreased from 0.39 mg/100ml to 0.03 mg/100ml with
0.1 %(w/v) tannin. Cd was also detected in the precipitate of treated fish sauce obtained by
centrifugation, and its amount corresponded to that of Cd removed from the treated sauce.
Total nitrogen, free amino acid and ACE inhibitor activity in the treated fish sauce were found
to be the same levels as those of untreated one. DPPH radical scavenging activitiy was reduced
by 28 %. Sensorial test did not show any difference between the treated and untreated fish
sauces.
0.5
Cd concentrations in fish sauce

0.4

0.3
(mg/100ml)

0.2

0.1

0.0
Untreated 0.01 0.05 0.1 0.2
Treated fish sauce with various conc. of tannin (% w/v)

Figure 1. Effect of added amounts of tannin on Cd concentrations in the treated fish sauces

CONCLUSION
Some fish sauce products prepared from squid contain certain amounts of Cd. We developed a
simple method to remove Cd from these products using tannin, one of the approved food
additives in Japan. The method consists of adding 0.1 % (w/v) tannin to fish sauce and
centrifuging out the resulting precipitate. Cd concentration (0. 39 mg/100ml) of the starting
fish sauce product decreased to 0.03 mg/100ml by this treatment. Chemical components such
as free amino acids and health beneficial functions present in fish sauce were found to be
retained in the tannin-treated fish sauce.

REFERENCES
[1] Jung-Nim, P., Fukumoto, Y., Fukumoto, Y., Fujita, E., Tanaka, T., Wahio, T., Otsuka, S., &
Watanabe, K. 2001. Chemical Composition of Fish Sauces Produced in Southeast and East Asian
Countries. Journal of Food Composition and Analysis, 14(2), 113-125.
[2] Nakazato, M., Tateishi, Y., Kobayashi, C., Yamajima, Y., Ohno, I., Kawai, Y., &Yasuda, K. 2000.
Hygienic Studies on Imported Fish Sause (II) -Contents of 9 Elements-. Ann. Rep. Tokyo Metr. Res.
Lab. P.H. 51, 155-159.
[3] Sakuta, Y., Nagano, N., Tomita, K., Wakasugi, M., Saitou, T., Shimakage, K., & Kitazaki, T. 2000.
Developments of Cadmium Removal Systems for Residues Wasted from Scallop Processing
Manufacture by an Electrochemical Method. Journal of the Japan Society of Waste Management
Experts, 11(3), 145-154.

1936
 Crispy air-dried pineapple rings: optimization of processing parameters

Giovanna Cortellino, Paola Pani, Danila Torreggiani

Research Unit of Food Technology - Council of Agricultural Research, Milan, Italy

(giovanna.cortellino@entecra.it)

INTRODUCTION
Traditionally, fruits in developing countries are sun dried and the quality of these products is
often poor. Osmotic dehydration, commonly used to remove part of the water content of fruit
before further drying, could improve sensory and functional properties. In particular
application of osmotic dehydration made better fruit texture and increased the stability of the
colour pigment. Great changes in the structure of vegetal tissue could be produced by both
osmotic dehydration and air drying [1]. In particular, texture of material moves from elastic-
visco-plastic to rigid, becoming fragile and brittle. These changes are welcomed when the final
product is a snack food such as pineapple rings, in which “crispy” and “crunchy” are sensory
attributes greatly influencing quality evaluation by the consumers [2]. The aim of this work is
to define the combined process, involving the osmodehydration and air-drying techniques, in
order to obtain dried and crispy rings of pineapple, having high qualitative characteristics.

MATERIALS & METHODS

Six mm thick pineapple rings were osmodehydrated for 30 minutes in pineapple juice and
sucrose solution, both at 50°Bx. Not pre-treated and pre-osmodehydrated rings were air dried
at 70-75-80 °C till constant weight. Dry matter, soluble refractometric residue, pH and total
tritable acidity of raw and osmodehydrated samples were measured. Solid gain and water loss
of the osmotic process were assessed. Dry matter and water activity of dried product were also
measured. Changes of colour due to processing were evaluated by image analysis technique.
Index crispness (Emod value) of final product was determined by bending snapping test [3]. The
sensorial characteristics were judged by a panel test.

RESULTS & DISCUSSION

During the osmotic process the initial dry matter and refractive index of pineapple rings
increased as a consequence of both water loss and solid gain regardless of osmotic solution
used. The osmodehydration in sucrose solution decreased significantly the pH of pineapple
rings. The same treatment in juice did not modify the pH while it caused a consistent increase
of the total acidity. The air dehydration process led to an evident browning of samples. This
was confirmed by higher a* values and lower L* and b* values of dried samples compared to
the raw ones. As expected increasing drying temperature colour was negatively influenced. The
osmotic pre-treatment in sucrose solution protected the colour during drying but the osmo-
dehydration with pineapple juice hadn’t the same positive influence.
The Emod values showed a positive progressive trend with the drying temperature in accordance
with the results obtained for osmo-air-dried apple rings [3]. The Emod values was also

11th International Congress on Engineering and Food - Athens, Greece, 2011 1937
influenced significantly by osmodehydration: lower values in both pre-treated samples than in
the not pre-treated ones.
As for sensorial tests the colour of samples dried at the lowest temperature was judged the
lightest and at the same time the most pleasant. By increasing the drying temperature the rings
acquired a colour tending towards orange/brown resulting less pleasant. The protective effect
of osmosis in sucrose solution wasn’t supported by the sensorial results. The flavour of the
samples dried at 75°C were judged more intense and pleasant if compared to those dried at the
other temperatures. Both osmosis treatments tended to improve the flavour and its
pleasantness. Among the samples dried at the same temperature the not pre-treated ones were
judged the crispiest, less firm and so more pleasant than the respective treated ones. According
to bending-snapping results the crispness, valuated by tasters, was positively influenced by the
increasing drying temperature. Overall the highest score for both crispness and pleasantness
was given to the not-treated samples dried at 75°C and 80°C.
Moisture content of dried pineapple rings decreased as the drying temperature increased
regardless of the pre-treatment applied, as verified for a similar apple product [3]. The lower
the dry matter the higher the water activity, which increased with diminishing of drying
temperature. At all the tested temperatures the dried rings had a significantly lower moisture
content and water activity than the respective osmo-air-dried ones. From both instrumental and
sensorial aspects, air-dehydrated samples resulted crispier than the respective osmo-air-
dehydrated ones independently of the solution type and temperature, confirming the inversely
proportional relationship between water activity and moisture content and crispness [3,4,5].

CONCLUSION
Contrarily to what was expected and reported by literature [3], the osmotic pre-treatment didn’t
contribute to the formation of the porous and crumbly structure of pineapple rings during the
drying process, whereas it contributed to the increase of the residual water content, which
affected the texture of final products. This fact supports the relevance of tissue characteristics
which influence osmotic exchanges and rheological behaviour. Nevertheless the osmotic
treatment in sucrose solution protected the colour during drying, confirming that the solid gain
limited the not enzymatic browning through air dehydration, as observed for other fruits. The
best result was obtained drying pineapple rings not pre-treated at 75°C: the product was
slightly amber-coloured, crispy and not too firm and consequently appreciated by the tasters.

REFERENCES
[1]. Lewicki P.P. 1998. Effect of pre-drying treatment, drying and rehydration on plant tissue properties:
a review. International Journal of Food Properties, 1, (1), 1-22.
[2]. Shewfelt R.L. 1999. What is food quality? Postharvest Biology and Technology, 15, 197-200.
[3]. Farris S., Gobbi S., Torreggiani D., Piergiovanni L. 2008. Assessment of two different rapid
compression tests for the evaluation of texture differences in osmo-air-dried apple rings. Journal of
Food Engineering, 88, 484-491.
[4]. Pittia P., Nicoli M.C., Sacchetti G. 2007. Effect of moisture and water activity on textural properties
of raw and roasted coffe beans. Journal of Texture Studies. 38, (1): 116-134.
[5]. Lewicki P.P., Marzec A., Kuropatwa M. 2007. Influence of water activity on texture of corn flakes.
Acta Agrophysica 9 (1): 79-90.

1938
 Extraction of polyphenols from grape seeds by unconventional methods and extract

concentration through polymeric membrane

Dan LIUa, Eugène VOROBIEVa*, Raphaëlle SAVOIREb, Jean-Louis LANOISELLÉa,c

a
Université de Technologie de Compiègne EA 4297 TIMR, Centre de Recherche de Royallieu, B.P.
20529, 60205 Compiègne, France (dan.ding@utc.fr) (eugene.vorobiev@utc.fr) (jean-
lous.lanoiselle@utc.fr)
b
Ecole supérieure de chimie organique et minérale EA 4297 TIMR, 1 allée du réseau J.M. Buckmaster,
60200 Compiègne, France (r.savoire@escom.fr)
c
Université de Bretagne Sud, BP92116, 56321 Lorient cedex, France

INTRODUCTION
Grape seeds are a by-product obtained after wine or juice making and present a good source of
functional compounds such as polyphenols, which are very valuable due to their antioxidant
capacity. Solvent extraction is the most commonly used technique for recovery of these
compounds [1]. However, the health concerns have sparked research into the safe extraction
protocols. Physical and electrical methods have been proposed as alternative methods.
Ultrasonication (US), pulsed electric field (PEF) and high voltage electrical discharge (HVED)
have been demonstrated to improve mass transfer in extraction of polyphenols from grapes [2].
The utilization of membrane technologies for concentrating and purifying phenolic compounds
from aqueous streams is a topic of growing interest. Membranes have been used to recover
phenolic compounds from extracts of mulberry root cortices, and to concentrate catechins from
aqueous grape stream [3].
The aim of this work was to apply the unconventional extraction methods including US,
HVED and PEF to extract polyphenols from grape seeds and concentrate the extracted
polyphenols by ultrafiltration.

MATERIALS & METHODS

HVED and PEF treatments are conducted with a high voltage pulse generator provided 40 kV-
10 kA discharges (Tomsk, Russia). The ultrasonic extraction is performed using an ultrasonic
processor (Hielscher GmbH, Stuttgart, Germany). The treatment is conducted with grape seeds
(50.0 ± 0.1 g) and distilled water (the liquid-solid ratio, L/S=5, w/w) at 50°C. The
concentration of polyphenols is determined by Folin–Ciocalteau method [4]. After the
treatment, samples are centrifuged for 10 min at 4000 g, and supernatant solution is collected
for dead-end filtration. PVDF membrane (50 kDa) is used and filtration is conducted with
volume concentration ratio (VCR) equal to 3.

RESULTS & DISCUSSION

Table 1 shows the results of physico-chemical properties of the samples after membrane
processing. It was shown that the quantity of polyphenols released by HVED is much higher
than the other two methods, which indicates that the HVED is a more effective method for the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1939
intracellular product extraction. After the filtration process, a remarkable reduction of the
polyphenol content is observed in the permeate fractions (57.0% for PEF, 61.5% for US and
78.3% for HVED, respectively). Consequently, polyphenols remain concentrated in the
retentate streams, which show a dark orange color. Considering the molecular weight of
polyphenols, this phenomenon can be explained by the screening effect of the membrane.

Table.1 Physico-chemical properties of feeds, permeates and retentates for three extraction methods
Polyphenol Clarity Color Conductivity Brix PH
g/L %T625 A420 ȝs/cm %
PEF
Feed 5.46 38.1 1.345 1185 1.9 4.66
Permeate 2.35 92.6 0.155 961 1.3 5.08
Retentate 10.31 4.98 4.9 1632 4.2 4.69
US
Feed 9.39 24.8 1.736 1252 2.2 4.70
Permeate 3.62 93.0 0.173 992 1.4 4.94
Retentate 21.68 2.27 5.6 1757 5.5 4.67
HVED
Feed 15.35 2.82 4.95 1795 4.9 5.15
Permeate 3.33 90.2 0.218 1013 2.1 5.24
Retentate 39.02 0.01 20.30 3090 10.7 5.00
T625 : clarity is characterized by transmittance at 625 nm
A420 : color is characterized by absorbance at 420 nm

CONCLUSION
HVED permits higher polyphenol extraction than the other two methods. During clarification,
high retention of polyphenols in the retentates is achieved. Electrically assisted technologies
combined with ultrafiltration demonstrated their effectiveness for the extraction and
concentration of polyphenols from grape seeds.

REFERENCES
[1] Lafka, T.I., Sinanoglou, V., & Lazos, E.S. 2007. On the extraction and antioxidant activity of phenolic
compounds from winery wastes. Food Chemistry, 104, 1206–1214.
[2] Boussetta, N., Lanoisellé, J.-L., Bedel-Cloutour, C., & Vorobiev, E. 2009. Extraction of soluble
matter from grape pomace by high voltage electrical discharges for polyphenol recovery: Effect of
sulphur dioxide and thermal treatments. Journal of Food Engineering, 95, 192–198.
[3] Díaz-Reinoso, B., Moure, A., Domínguez, H., & Parajo, J.C. 2009. Ultra- and nanofiltration of
aqueous extracts from distilled fermented grape pomace. Journal of Food Engineering, 91, 587–593.
[4] Ribéreau-Gayon, P., Sudraud, P., Milhé, J.C., & Canbas, A. 1970. Recherches technologiques sur les
composés phénoliques des vins rouges. Connaissance Vigne Vin 2, 133–143

1940
 Performance of bovine and ovine liquid whey protein concentrate on functional

properties of set yoghurts

Marta Henriquesa,b, David Gomesa, Daniela Rodriguesa, Carlos Pereiraa, Maria Gilb

a
Escola Superior Agrária - Department of Food Science and Technology, Polytechnic Institute of
Coimbra, Coimbra, Portugal (mhenriques@esac.pt; david@esac.pt; cpereira@esac.pt)
b
CIEPQPF, Chemical Engineering Department, Faculty of Science and Technology, University of
Coimbra, Coimbra, Portugal (hgil@eq.uc.pt)

INTRODUCTION
Increasing protein content in yoghurts implies the milk fortification by the addition of SMP.
More recently, whey protein concentrate (WPC) are also used due to their availability and low
cost. Despite WPC are largely applied as attractive food ingredient in a wide range of food
applications, the direct reincorporation of liquid whey protein concentrates (LWPC) in dairy
products, being a less expensive alternative, is seldom referred. The effect of the replacement
of SMP by WPC on textural and physicochemical properties of yoghurts has been reported by
several authors but in some cases their conclusions were contradictory. The reasons pointed for
that are the significant variations in the functionality of WPC resultant from the whey
processing conditions, specially heating and the whey source. Based on the bovine and ovine
cheese production, in Portugal, the overall volume of whey produced annually is estimated in
approximately 560 000 tonnes. The use of membrane technology, namely ultrafiltration (UF)
enables the extraction and concentration of whey proteins from whey to its reincorporation in
production, solving environmental problems and adds value to existing products.
No information is available about the LWPC functionality in yoghurt. In this work we intended
to evaluate the effects of partial substitution of SMP by LWPC of bovine and ovine origin on
physicochemical, textural, rheological and sensorial properties of set yoghurts as well as to test
the acceptability of ovine yoghurt as an alternative product in Portugal.

MATERIALS & METHODS

LWPC production consisted in the concentration of whey at 30ºC in a batch ultrafiltration pilot
plant. The resulting retentate suffered a thermal treatment (90ºC/5min) followed by
homogenization at 100 bar. After LWPC characterization the product was incorporated in milk
batches used for yoghurt production. Four yoghurt formulations with 16% total solids were
produced. Ovine yoghurts (LO) were produced exclusively with ovine skimmed milk. The
formulations with bovine milk were normalized in fat content with cream, and protein content
using respectively: (i) SMP (conventional bovine yoghurt (LB)); (ii) 7.3% of bovine LWPC +
4.4% of SMP (LB-LWPCb) and (iii) 7.3% of ovine LWPC + 4.8% of SMP (LB-LWPCo).

RESULTS & DISCUSSION

Comparing conventional yoghurts with the ones produced with LWPC incorporation, no
significative differences in color were found, only a decreased in L* value during storage.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1941
The textural analysis did not differ over the products shelf live in each type of formulation,
neither between formulations for cohesiveness, springiness and resilience. Hardness,
adhesiveness and gumminess were significantly higher (p<0.05) for the ovine yoghurts (LO),
followed by the bovine yoghurts (LB) and finally by the products with incorporation of LWPC.
The high amount of solids in tested formulations is in part, responsible for the low syneresis
indexes observed (0.5-5.0%). However, the use of LWPC decreased water holding capacity by
increasing yoghurts syneresis. This behaviour can be explained by the nature, proportion and
incorporation form of proteins in each formulation (mainly caseins in conventional products
and denaturated whey proteins in tested ones). Yoghurt viscosity varied inversely with
syneresis, allowing the identification of three distinct groups of products. The first group
includes ovine yoghurts (LO) and it is characterized by low values of syneresis and higher
viscosity. Such behaviour can be explained by their higher protein concentration and therefore
the possibility of building a more cohesive polymer network. Set yoghurts prepared with
LWPC exhibited the lowest values in viscosity and the highest syneresis index. The last group
includes LB yoghurts which present intermediate values for both parameters. Although the
total protein content between the last two groups of yoghurts does not differ significantly, the
protein’s nature proportion in the formulations (casein and whey proteins) was distinct, and
pointed as the responsible for these differences [1]. Our results are also in accordance with data
published by Modler et al. (1983) and Sodini et al. (2005) who concluded that products
enriched with SMP or higher casein contents tend to produce more compact and, more viscous
gels with higher retention capacity then the ones enriched with whey proteins. During storage,
no particular trend in both properties was observed.
Despite the similarity in biochemical composition between LO and LB yoghurts the sensory
panel differentiated both products (p<0.001). The yoghurts with the incorporation of LWPC
were the preferred ones and the ovine yoghurts (LO) the less appreciated. These results showed
that the yoghurts with closer textural properties, viscosity and syneresis to the conventional
ones (LB) were more appreciated. The lake of familiarization with the ovine yoghurts by
Portuguese consumers reflects the lower acceptance of these products.

CONCLUSION
It was concluded that partial substitution of conventional SMP by LWPC in set yoghurts
(independently of their origin) is possible and can be very attractive not only concerning to the
global process yield, as well as by reducing effluents and adding value to the existing products,
but also in what concerns to their functional properties. Further work envisages the
optimization of the LWPC denaturation step, as well as on the improvement of sensory
properties of ovine milk yoghurts.

REFERENCES
[1] Guzmán-González M. Morais F. Ramos M. & Amigo L. 1999. Influence of SMC replacement by dry
dairy products in a low fat set-type yoghurt model system. I: Use of WPC, MPC and SMP. J Science
Food Agric., 79, 1117-1122.
[2] Modler H.W. & Kalab M. 1983. Microestructure of yogurt stabilized with milk proteins. J Dairy
Science, 63, 430-437.
[3] Sodini I. Montella J. & Tong P.S. 2005. Physical properties of yogurt fortified with various
commercial whey protein concentrates. J Science Food Agriculture, 85, 853-859.

1942
 Manufacture of Gelatin-Based Films Using Extrusion: Assessment of Extrusion

Parameters on Film Properties

Z. A. Nur Hanani, E. Beatty, Y. H. Roos & J. P. Kerry

Food Packaging Group, School of Food & Nutritional Sciences, University College Cork (UCC), Cork,
Ireland (Joe.Kerry@ucc.ie)

INTRODUCTION
Biopolymers have a potential role to replace some plastic materials in industry, particularly for
certain food packaging applications. Gelatin is a protein-based polymer, commercially obtained
from the skins and bones of cattle and pigs during slaughtering. Gelatin has gained more
attention as an ingredient capable of producing edible or biodegradable films because of its
abundance and usage costs. However, fish gelatin has been investigated as an alternative to
mammalian gelatin usage within the food industry. Fish gelatins have demonstrated suitable
film-forming abilities, yielding transparent and highly deformable films. Fish gelatin cannot
replace the role of mammalian gelatin in many applications because fish gelatin has a low
melting point and low gel strength, but with a relatively high viscosity, compared to
mammalian gelatins. However, its potential role in film manufacture deserves to be
investigated. From the scientific literature, gelatin films for biodegradable food packaging
applications have been predominantly prepared using casting methods. However, extrusion is
one of the most important processing techniques required to produce plastics on a commercial
scale [1]. Thus, in this study, fish-derived gelatine was used to manufacture films via the use of
extrusion. The objective of this study was to evaluate the effect of modifying selected extrusion
parameters on the physical properties of the resulting films.

MATERIALS & METHODS

Gelatin films from fish skin (Healan Ingredients Ltd, York, UK) were prepared by twin screw
co-rotating extruder (APV Baker, Peterborough, UK). The barrel had 4 heating zones with a
screw diameter of 19 mm and length to diameter ratio (L/D) of 25:1. First, the temperatures of
the four heating zones were all set at 80 °C and the screw speed was adjusted from 100 to 200
rpm. Then the screw speed was fixed to 150 rpm and the temperatures of the 4 heating zones
were adjusted.

RESULTS & DISCUSSION

From the colour measurement, the lightness (L values) of the films significantly (P 0.05)
decreased when the screw speed of extruder increased. The increase of screw speed
significantly (P 0.05) increased film yellowness (b values). The increase in screw speed
increased the amount of shear applied to the gelatine, thereby promoting discoloration [2]. The
barrel temperature also affected the lightness and yellowness values for manufactured films.
Mechanical properties of fish gelatin films are shown in Table 1. The extrusion process
significantly improved the mechanical properties of fish gelatin films. TS values for gelatin
films significantly (P 0.05) increased when the speed of extruder also increased. Improved
TS values using higher screw speeds can be attributed to enhanced dispersion of gelatin

11th International Congress on Engineering and Food - Athens, Greece, 2011 1943
powder. Kumar et al. [1] reported the same trend for soy protein-based films when extrusion
screw speeds increased. PS also significantly (P 0.05) increased with increasing screw
speed. However, the effect of screw speed on EBP was not significant. Comparing films heated
at different temperatures, it was noted that the use of a higher temperature contributed to lower
TS values. The effect of temperature on PS was not significant. However, EBP values
significantly (P 0.05) increased with increasing temperature application. It can be concluded
that higher screw speeds are required to prepare films with increased mechanical properties.

Table 1. Mechanical properties of gelatin films

Thickness Tensile strength Puncture strength Elongation

Samples (ȝm) (MPa) (N) (%)
Speed
100 0.065 ± 0.016a 5.64 ± 0.50bc 3.36 ± 0.23c 3.65 ± 0.36b
150 0.042 ± 0.015b 6.45 ± 0.15b 5.57 ± 0.47b 4.40 ± 0.29ab
200 0.035 ± 0.006b 9.11 ± 2.05a 6.80 ± 0.33b 4.89 ± 0.93a
Temperature
65, 75, 85, 95 0.029 ± 0.006b 4.28 ± 0.45a 10.51 ± 1.84a 3.61 ± 0.31b
65, 85, 105, 85 0.047 ± 0.007ab 1.99 ± 0.20cde 10.63 ± 1.68a 5.13 ± 0.63a

Means in the same column followed by the same letter are not significantly different (P 0.05).

Analysis of variance using the general linear model showed that effect of screw speed and
temperature on WVP was not significant. According to FTIR results, all the spectra obtained
had no changes in vibrational wavenumber except for amide-A peak. FTIR has shown that the
wavenumber of amide-A has shifted higher when the screw speed increased. This is possibly
due to lower formation of hydrogen bonding. Conversely, the wavelength of amide-A is
decreased when a higher barrel temperature was used. According to Jongjareorak et al. [3],
this structural alteration might be related to the mechanical properties of films. However,
further studies are required to investigate these results.

CONCLUSION
Fish gelatine-based films were successfully produced using extrusion. The parameters of
extrusion process have an important influence on the final structure and properties of such
films.

REFERENCES
[1] Kumar, P., Sandeep, K. P., Alavi, S., Truong, V. D. & Gorga, R. E. 2010. Preparation and
characterization of bio-nanocomposite films based on soy protein isolate and montmorillonite using
melt extrusion. Journal of Food Engineering, 100, 480-489.
[2] Berset, C. 1989. Color. In: Mercier, C., Linko, P. & Harper, J. M. Extrusion Cooking. American
Association of Cereal Chemists, Inc. Minnesota, USA.
[3] Jongjareonrak, A., benjakul, S., Visessanguan, W., Tanaka, M., 2008. Antioxidative activity and
properties of fish skin gelatin films incorporated with BHT and Į-tocopherol. Food hydrocolloids, 22,
449-458.

1944
 Combining microwave and jet-impingement in a oven prototype

Gianpaolo Ruoccoa, Maria V. De Bonisa, Francesco Marrab

a
University of Basilicata, College of Food Technology, Potenza, Italy (gianpaolo.ruocco@unibas.it)
b
University of Salerno, Department of Industrial Engineering, Fisciano (SA), Italy (fmarra@unisa.it)

INTRODUCTION
Although the use of microwave (MW) ovens has been proven beneficial in food preparation due to its
speed and convenience, the potential of MW heating is currently not fully realized due to its generally
non-uniform effects on the product finishing. A viable option to mitigate this is the use of an optimized
forced convection, as the jet impingement (JI), that result in improved heat and mass transfer, even for
moderately turbulent flows [2]. In the research proposed herein a MW/JI combination oven prototype has
been set-up and operated, exercising low MW power density, also to prevent pressure build-up and
splatter and allow monitoring and calibration [3,4].

MATERIALS & METHODS

The experiments have been carried out in a prototype plant consisting of a commercial MW oven, with 5
nominal power levels up to 1000 W, and an auxiliary system of JI heating. The oven was modified by
introducing a calibration nozzle to realize a precise outlet velocity profile of the jet [1].

RESULTS & DISCUSSION

The analysis is focussed upon the dimensionless descriptors for the fluid dynamic (Re) and local thermal
(T*) regimes, and process duration (ǻt*). Potato samples were placed on a wood mesh pedestal, directly
under the injected air. Three optical thermometers continuously read T*1, T*2 and T*3 (for the top, centre
and bottom locations, respectively) at the sample's vertical axis. A total of 10 operating schemes were
employed by varying MW intensity and jet velocity and temperature: 1) pure MW treatments, with
operating schemes MW2-JI0 (337 W, nominal) and MW3-JI0 (555 W, nominal); 2) combined MW/JI
treatments, i.e. enhanced by the application of JI obtained by varying the TJ and Re: MW2-JI1, nominal
MW power 337 W, with Re=8.86×103 and TJ=330 K; MW2-JI2, nominal MW power 337 W, with
Re=8.09×103 and TJ=373 K; MW2-JI3, nominal MW power 337 W, with Re=14.5×103 and TJ=330 K;
MW2-JI4, nominal MW power 337 W, with Re=13.3×103 and TJ=373 K; MW3-JI1, nominal MW power
555 W, with Re=8.86×103 and TJ=330 K; MW3-JI2, nominal MW power 555 W, with Re=8.09×103 and
TJ=373 K; MW3-JI3, nominal MW power 555 W, with Re=14.5×103 and TJ=330 K; MW3-JI4, nominal
MW power 555 W, with Re=13.3×103 and TJ=373 K.
The nominal MW power was determined by exposure times, that were regulated by imposing a
magnetron working time ǻt*1, followed by a resting or relaxation period ǻt*2, in which the heat ad mass
transport in the subject sample is residual only.
In MW2 cases, with the increasing Re, the heating on-set loses synchrony with the adopted MW
production mode in all sample’s location, as the internal conduction is more effective instead. At the top
location (T*1), comparing the progress for MW2-JI4 to that for MW2-JI2 in Figure 1, the increment of
40% of Re results after ǻt*=2 in about a 30% increment in the dimensionless temperature. This effect is
milder, as expected, for a colder JI (for MW2-JI3). At the centre location (T*2) the increment of the
convective mechanism is barely felt, so the heating relies more on conduction. At the bottom location
(T*3) the aforementioned cooling effect is more evident, for increasing Re, even for warmer JI.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1945
In all the position of MW3 cases, the pure MW process allows for higher temperature. As in previous
cases, among investigated positions, the bottom is the one exhibiting the higher temperatures, the top
position being the colder. When JI is combined to MW, in all the investigated positions the reached
temperatures are lower with respect to pure MW exposition and the behaviour of heating in the
investigated positions is the opposite: the top exhibits the higher temperature values while the bottom
results as the cooler position. The cooling effects of JI, at higher Re number, are due by its capability to
convey the moisture far from the sample thus cooling it down. The higher cooling effects at samples
bottom are probably due by that accumulation of moisture in the lower part of the sample, as discussed in
the above sections. In the present case, the MW power being the higher among considered case, the
sample structure is subjected to stronger electromagnetic field: a porous structure is formed as result of
the intense heating and moisture easy goes to bottom enhancing, locally, the capability to further absorb
MW power. But the wetter the bottom, the better the JI exerts its convective action on the moisture, thus
taking away energy as latent heat.

Figure 1. Thermal histories in the sample measurement locations for the MW2-JI0, MW2-JI1, MW2-JI2 treatments
(top left), for the MW2-JI0, MW2-JI3, MW2-JI4 treatments (top right), for the MW3-JI0, MW3-JI1, MW3-JI2
treatments (bottmo left) and for the MW3-JI0, MW3-JI3, MW3-JI4 treatments (bottom right).

CONCLUSIONS
An experimental work has been proposed on a lab-scale plant to combine MW exposure and localized
forced convection. For Re higher than 13000 a cooling effect is detected even for hot jets, whereas for
milder flows the action of jet impingement is almost irrelevant. For long microwave exposures, an excess
of moisture is found at the sample’s bottom, therefore the heating increases due to the greater energy
absorption, and the jet impingement may lead to strong localized cooling.

REFERENCES
[1] Angioletti, M., Di Tommaso, R.M., Nino, E. & Ruocco, G. 2003. Simultaneous visualization of flow field and
evaluation of local heat transfer by transitional impinging jets. International Journal of Heat and Mass Transfer, 46,
1703-1713. [2] De Bonis, M.V. & Ruocco, G. 2007. Modelling local heat and mass transfer in food slabs due to air jet
impingement, Journal of Food Engineering, 78, 230-237. [3] Marra, F., Zhang, L. & Lyng, J.G. 2009. Radio frequency
treatment of foods: review of recent advances. Journal of Food Engineering, 91(4), 497-508. [4] Marra, F., De Bonis,
M.V. & Ruocco, G. 2010. Combined microwaves and forced convection heating: a conjugate approach. Journal of
Food Engineering, 97(1) 31-39.

1946
 The Sequential Ventilation of Cheese Ripening Rooms: an Eco-Design Approach?

Pierre-Sylvain Miradea, Bruno Perretb, Hervé Guilleminb, Daniel Picqueb, Cécile Callonc, Marie-Christine
Montelc, Georges Corrieub

a
UR370 Qualité des Produits Animaux, INRA, 63122 Saint-Genès-Champanelle, France
(e-mail: pierre-sylvain.mirade@clermont.inra.fr)
b
UMR782 Génie et Microbiologie des Procédés Alimentaires, INRA, 78850 Thiverval, France
c
UR545 Recherches Fromagères, INRA, 20 Côte de Reyne, 15000 Aurillac, France

INTRODUCTION
Part of the integrated European ‘Truefood’ project was dedicated to studying ripening room
management strategies that aimed at reducing energy costs, particularly by adopting sequential
ventilation systems. In a first study [1], results obtained on two types of cheeses showed that
significant improvements could be achieved in terms of cheese ripening room monitoring and
control using sequential air ventilation in small-sized ripening rooms. The objectives of this
‘industrial-scale’ study were to test a new ripening room system based on sequential air
ventilation, and to determine the savings in terms of electrical energy consumption resulting
from these new ventilation conditions.

MATERIALS & METHODS

The industrial ripening room selected in this study is currently used to ripen traditional PDO
Saint-Nectaire cheeses. We designed and produced a lab prototype integrating the most
significant measurements (air temperature, RH, CO2 and O2 contents, electrical energy
consumption) and automatically controlling ripening room conditions. Furthermore, in-house
software was specially adapted for monitoring this industrial prototype and used to control air
ventilation regime, i.e. either continuous or sequential, and either time-controlled or
temperature-controlled. Over one year, six different cheese ripening trials were performed: (1)
two reference trials, called Ref1 and Ref2, with continuous air ventilation, (2) two trials with
“time-based” sequential ventilation (SV50 and SV60), and (3) two trials with “temperature-
based” sequential ventilation, called SVRT1 and SVRT2. Each of these six trials was led for 28
days. Two cheeses were sampled on day 1 and then every week to analyze the microbial,
biochemical and sensory evolution.

RESULTS & DISCUSSION

The effect of sequential air ventilation in the ripening room was assessed through 3 main
factors: cheese respiratory activity, energy consumption, and evolution of Saint-Nectaire
ripening dynamics and final product quality.
Regarding the first factor, measurements indicated that cheese ripening created significant
respiratory activity. This activity was not significantly affected by ripening room ventilation
regime, whether continuous or sequential. Air renewal and door opening were identified as the
two most important factors acting on the amount of CO2 and O2 measured in the room.
Regarding energy consumption, preliminary measurements showed that electrical energy
consumption in the room was essentially due to ventilation. Consequently, sequential

11th International Congress on Engineering and Food - Athens, Greece, 2011 1947
ventilation led to significant savings compared to the continuous ventilation routinely used in
ripening rooms. The data from each of the 6 trials performed was used to determine mean
electrical energy consumptions and savings listed in Table 1. On this basis, an improved
approach to ripening room monitoring was suggested, which entailed temperature-based
sequential ventilation, with a temperature variation ideally ranging from 0.6 to 0.8°C, together
with disconnecting the room heating device. The economic impact of the resulting electricity
savings culminated at 0.55% of the productivity of the cheese ripening process.

Table 1. Savings on electrical energy consumption obtained with sequential ventilation and cutting the
heating. CV+H: continuous ventilation with heating; CV-H: continuous ventilation without heating; SV:
time-based sequential ventilation; SVRT: temperature-based sequential ventilation; ǻT: temperature
difference between the low and high set-points
Trial Working conditions Electrical energy Daily economy Percentage gain (%)
consumption (kWh)
(kW/d) H SV H SV
Ref1 CV+H 253 0 0 0
Ref2 CV-H 212 41 0 16 0
SV50 SV 109 41 103 16 49
SV60 SV 85 41 127 16 60
SVRT1 SV ǻT=1°C 83 41 129 16 61
SVRT2 SV ǻT=0.7°C 87 41 125 16 59
SVRT2 SV ǻT=0.3°C 85 41 127 16 60
SVRT2 SV ǻT=0.3°C 164 41 48 16 23
without blowing ducts

To compare the evolution in cheese ripening dynamics and final quality as a function of
ventilation conditions, we evaluated the main microbiological, biochemical and sensory
characteristics of the cheeses. Results showed that sequential ventilation had no significant
effect on cheese quality. Ripening microbial flora, the main biological characteristics of the
cheese, and sensory descriptors showed little evolution versus ripening time compared to
continuous ventilation.

CONCLUSION
Under the conditions of this study, mean savings in electrical energy consumption under
temperature-based sequential ventilation at a temperature variation close to 0.7°C reached
125 kWh per day, which translates into a 0.43% increase in ripening room productivity. It was
proposed to further improve the ripening room by disconnecting the room heating device in
order to keep heat production down at the level provided by the cheese respiration activity.
Finally, it was shown that sequential ventilation had no significant effect on cheese quality.

REFERENCE
[1] Picque D., Guillemin H., Mirade P.S., Didienne R., Lavigne R., Perret B., Montel M.C. & Corrieu G.
2009. Effect of Sequential Ventilation on Cheese Ripening and Energy Consumption in Pilot
Ripening Rooms. International Dairy Journal, 19, 489-497.

1948
 Influence of additives on white tin loaf alveolloli formation

Cecília Dominguesa, Paula Prazeresb, Paula Correiaa,c

a
Escola Superior Agrária, Instituto Politécnico de Viseu, Viseu, Portugal
(raquel.domingues@hotmail.com)
b
Bimbo, Produtos Alimentares. Albergaria-a-Velha. Portugal (paularita.prazeres@saralee.com)
c
CI&DETS, Instituto Politécnico de Viseu, Viseu, Portugal(paulacorreia@esav.ipv.pt)

INTRODUCTION
Bread has always been one of the most popular and appealing food products due to its superior
nutritional, sensorial and textural characteristics, ready to eat convenience as well as cost
competitiveness. Bread is essential to the diets of many people worldwide. The ingredients of
bread will impart characteristic colours, texture, and nutritional value which may improve
quality of the bread. Therefore, a proper balance of ingredients needs to be achieved to produce
high-quality bread. Concerns about the quality of bread go beyond the ingredients in the loaves
themselves. One of the main quality criteria of bread is related with texture, and the
development of a desirable volume, related to alveolloli formation. In white tin loaf it is also
important that these alveolloli have a diameter < 2 mm, to ensure uniformity throughout the
bread. Gluten and monocalcium phosphate are involved in the fermentation process, the first
being a leavening agent that causes baked goods to rise. The objective of the present study was
to produce a white tin loaf with different additive formulations to evaluate their effect on the
alveolloli formation and determine which amount of additives is the optimum for a uniform
distribution.

MATERIALS & METHODS

White tin loaves were made at the laboratory, simulating equal conditions to the ones used on
an industrial scale. A Central Composite Rotatable Design (CCRD) of the experiment was
applied, using the Response Surface Methodology (RSM). The experiments were performed
with k = 2 for: additive (monocalcium phosphate and gluten) as independent variables. The
legal addition limit in Portugal for monocalcium phosphate is 10g to one bread kilogram (DL
121/98, 8th May). The upper and lower limits for these variables were established by national
bread legislation and enterprise rules. The dependent variables included total alveolloli
number, number of alveolloli with a diameter t 2 mm and number of alveolloli with a diameter
< 2 mm. For the alveolloli measurements it was use the “Image J” software, which is a public
domain, Java-based image processing program (National Institutes of Health, USA).

RESULTS & DISCUSSION

The results of the analysis of variance obtained with the white tin loaf made with different
additive formulations to evaluate their effect on the total alveolloli number, number of
alveolloli with a diameter t 2 mm and number of alveolloli with a diameter < 2 mm fit to the
model proposed. In spite of this, RSM regression equations did not adjust to the dependent
variable number of alveolloli with a diameter t 2 mm, which presented a R2 = 0.3. A strong

11th International Congress on Engineering and Food - Athens, Greece, 2011 1949
correlation was found between the total number of alveolloli and number of alveolloli with a
diameter < 2 mm (r = 0.965), and this as a refection on the RSM results, which presented
similar responses. Thus, Figure 1 shows that the number of alveolloli is higher for low
concentrations of monocalcium phosphate/ gluten, which means that to produce a white tin loaf
with a high number of alveolloli with a diameter < 2 mm the quantity of monocalcium
phosphate and gluten must be lower than 0.8g and 0.4g, respectively. The results also revealed
that without any addition of these additives a white tin loaf will also present uniform alveolar
formation.

300
250
200
150
100
50

Z=309.31774-45.64590*MP-86.34899*MP2-236.73591*G+58.22195*G2+61.21212*MP*G ; R2 = 0.88

Figure 1. Number of alveolloli with a diameter < 2 mm response surfaces and adjusted
regression equation for white tin loaf produced with addition of monocalcium phosphate (MP)
and gluten (G).

CONCLUSION
The present work allows a deeper knowledge about the implication of addition of gluten and
monocalcium phosphate on the alveolloli formation in industrial white tin loaves, which is
closely related with bread quality. It was found a linear correlation between the mean of total
alveolloli and the mean of alveolloli with diameter < 2 mm. Results showed that to obtain
bread with a higher number of alveolloli the optimum quantities of monocalcium phosphate
and gluten added should be less than 6 g/ Kg bread and 3 g/ kg bread, respectively. It was also
possible to conclude that if white tin loaf is produced without monocalcium phosphate and
gluten it will also present a uniform alveolar formation.

1950
 Textural properties of vegetables: a key parameter on ultrasonic assisted convective

drying

C. Ozuna, J. A. Cárcel, J. V. Santacatalina, A. Mulet and J. V. García-Pérez*

Grupo de Análisis y Simulación de Procesos Agroalimentarios. Departamento de Tecnología de Alimentos.

Universidad Politécnica de Valencia. Camino de Vera. s/n. E46022.
Valencia, Spain (jogarpe4@tal.upv.es)

INTRODUCTION
The increasing need for producing high quality dried products has led to combine traditional drying
methods with non-conventional energy sources. In this sense, ultrasonic energy is very promising because
it can act without affecting the main characteristics and quality of products due to its low heating effects.
Product characteristics also influence the absorption of ultrasound; so far only porosity has been
addressed [1]. Therefore, it should be expected that the ultrasonic effects on the material being dried will
be linked to product’s textural properties. Textural properties of vegetable tissue are related with
histological factors, such as, size, shape, adhesion, intercellular spaces, wall properties and turgor
pressure of the cells. For evaluating textural properties on vegetables it is essential to understand their
mechanical behavior in terms of different rheological parameters [2]. In this sense, texture profile analysis
(TPA) is a method commonly used to evaluate mechanical properties on this kind of products [3]. The
aim of this work was to identify the influence of the textural properties of different vegetable products on
the ultrasonic assisted convective drying effectiveness.

MATERIALS & METHODS

Experimental drying kinetics (40 ºC and 1 m/s) of different vegetable products already reported in
previous works were used: carrots and potato cubes (8.7 mm), eggplant cylinders (height 20 mm and
diameter 24 mm), orange and lemon peel slabs (thickness 5.95 ± 0.41 mm). For each case, different
ultrasonic powers (UP): 0, 6, 12, 18, 25, 30 and 37 kW/m3 were applied until samples lost 70 % of the
initial weight. Drying kinetics were modeled applying a diffusion model which neglected the external
resistance to mass transfer. The effective moisture diffusivity was identified by using an optimization
procedure, the Generalized Reduced Gradient (GRG), available in Microsoft ExcelTM spreadsheet from
MS Office 2007. The goodness of the fit was determined by calculating the percentage of explained
variance (%VAR). The multifactor ANOVA and the LSD intervals were chosen to evaluate the
significance (p<0.05) of the differences between the effective diffusivity values identified. Textural
properties of the different products were assessed by TPA analysis. In the case of orange and lemon peel,
the analysis was carried out in both faces of the albedo and flavedo tissue and averaged. Hardness,
adhesiveness, cohesiveness, springiness, chewiness and resilience were calculated from force/deformation
profiles [2]. At least, 10 measurements were performed for each set of samples.

RESULTS & DISCUSSION

The applied ultrasonic power during drying showed a significant (p<0.05) influence on the effective
moisture diffusivities. The ultrasonic effects were dependent on the applied power, the higher the power,
the greater the identified effective diffusivity. For the different products tested, linear relationships
between the ultrasonic power level and the effective moisture diffusivity were found in the power range

11th International Congress on Engineering and Food - Athens, Greece, 2011 1951
tested in this work (0-37 kW/m3). The improvement of the De values may be associated with the
mechanical effects brought about by applying ultrasound to the material being dried.[1] The proposed
diffusion model provided low percentages of explained variance (< 90 %). In order to obtain a better fit of
experimental data, the hypothesis assumed to solve the diffusion equations should be reconsidered.
The main differences for the products tested in terms of texture were observed in hardness and chewiness
values. Carrot and potato showed the higher values. The relatively high volume of cell wall material and
strong cohesion in these products contributes to the higher values of hardness and chewiness than
eggplant, lemon and orange peel. Therefore, citrus peels and eggplant may be considered non-compact
structures with a low mechanical resistance to deformation.
0.25

Ɣ Carrot
Ƈ Eggplant
0.20
x Lemon peel
Ŷ Orange peel
Slope (m5 /kJ)

ŸPot ato
0.15

0.10

y = -0.0063x + 0.2681
R² = 0.92
0.05

0.00
0 10 20 30 40 50

Hardness (N)

Figure 1. Influence of the hardness on the acoustic effectiveness on the drying process.

As can be observed in Figure 1, a linear relationship between the slopes of the linear relationships of the
effective diffusivity and the acoustic power versus hardness was found, the higher the hardness of the
tested products, the lower the slope. That means that the improvement due to ultrasound effects on the
effective diffusivity is well correlated (at 99 % confidence level) with the hardness of the product. This
fact confirms that the mechanical compressions and expansions (“sponge effect”) produced in the
materials by ultrasound were more intense in soft products resulting in a more effective water removal.
Furthermore, the acoustic effects on boundary layer of intercellular spaces could be also more intense in
this type of products due to a larger porous net. In the case of the chewiness parameter, a similar behavior
was observed than that found for the hardness. However, other texture parameters calculated
(cohesiveness, springiness, adhesiveness and resilience) were not well correlated with the effect of
ultrasound.

CONCLUSIONS
According to the results of this work, the ultrasonic effectiveness on the drying process depends on the
structure of the material being treated. A preliminary instrumental textural analysis could be used to
evaluate the potential of the ultrasonic application for the drying of a particular product.

REFERENCES
[1] García-Pérez J.V., Cárcel J.A, Riera E. & Mulet A. 2009. Influence of applied acoustic energy on the drying of
carrots and lemon peel. Drying Technology, 27, 281-287. [2] Ferreira D., Lopes da Silva A., Pinto G., Santos C.,
Delgadillo I. & Coimbra M. A. 2008. Effect of sun-drying on microstructure and texture of S. Bartolomeu pears (Pyrus
communis L.). European Food Research Technology, 226, 1545-1552. [3] Alvarez M.D., Canet W. & López M.E.
2002. Influence of deformation rate and degree of compression on textural parameters of potato and apple tissues in
texture profile analysis. European Food Research Technology. 215, 13-20.

1952
 The influence of palm oil quality on the refining conditions

Klicia A. Sampaioa,*, Jose V. Ayallab, Simone M. Silvaa, Roberta Cerianic, Roland Verhéd and Antonio J.
A. Meirellesa

a
EXTRAE, Food Engineering Faculty, University of Campinas, Zip Code, 13083-862, Campinas, São
Paulo, Brazil (klicia@fea.unicamp.br)*
b
Desmet & Ballestra R&D Center, Zip Code, 1935, Zaventem, Belgium
(jose.vila.ayala@desmetballestra.com)
c
School of Chemical Engineering, University of Campinas, Zip Code, 13083-852, Campinas, São Paulo,
Brazil (rceriani@feq.unicamp.br)
d
Faculty of Bioscience Engineering, Ghent University, Zip Code, 9000, Ghent –Belgium
(roland.verhe@ugent.be)

INTRODUCTION
The quality of any crude oil is to be considered as it can greatly affect the efficiency of the refining
process and the quality of the end-products. The most used parameters to asses crude palm oil
quality is: FFA; phosphatide content; peroxide value (PV); metal traces and the deterioration of
bleachability index (DOBI). Although, the FFA is one of the most frequently determined quality
indices during the production, storage and market of palm oil products and the oil price is dictated
by FFA content. The reported range of free fatty acid content of crude palm oil was 2.3–6.7% [1].
Edible oils are refined in order to remove unacceptable materials with the least possible loss of oil
and without affecting desirable compounds that are present in the oil in small quantities [2].
Refining of palm oil is preferably performed by physical means, since its high acidity can lead to
excessive losses of neutral oil in case of the caustic refining process [3].
For multivariable processes such as food systems, a large number of factors can influence the
responses of the process [4]. Thus, the aim of this work was to verify the effects of two operating
parameters i.e., temperature (ranging from 200 °C to 260°C) and steam flow (from 0.5% to 3.5%),
and one quality parameter, i.e., the initial oil acidity (from 2.22% to 6%) in the removal of free fatty
acids and loss of neutral oil, following a 23 factorial design.

MATERIALS & METHODS

Crude palm oil (CPO) was kindly provided by the industry (Agropalma S/A, Amazon Refining
Company, Brazil). The oil was then submitted to regular degumming and bleaching process.
Commercial oleic acid (MERCK, USA) was added to adjust the acidity of each model system.
The experiments were carried out using a 23 full-factorial design with three central points to
estimate the reproducibility of the trials. The steam deacidification experiments were carried out in a
laboratory-scale batch deodorizer (Desmet & Ballestra – Zaventem, Belgium). The glass batch
deodorizer was loaded with bleached palm oil and placed in an oven with controlled and monitored
temperature.

RESULTS & DISCUSSION

The experimental results (Table 1) revealed that the final FFA and the NOL were a function of the
three variables studied. However, the effect of the temperature was predominant in both responses,
followed by the steam percentage.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1953
Comparing trials 9 and 10, it is possible to verify the effect of increasing temperature on the
decrease in the final FFA and on the increase in NOL, due to volatilization of fatty compounds at
higher temperatures. A comparison of trials 11 and 12 showed that an increase in the steam
percentage, promoted a decrease in the final oil acidity and an increase in the neutral oil loss, since
it reduced the required partial pressure of the volatiles. The effect of increasing the initial oil acidity
on the removal of FFA and on the NOL can be seen by comparing trials 13 and 14.

Table 1. Central composite design experiment data for the physical refining of palm oil
Real Variables Responses
Trial
T (°C) Steam (%) IOA (%) Final FFA (%)a NOL (%)b
1 212.1 1.11 2.99 0.455±0.001 0.062
2 247.9 1.11 2.99 0.112±0.013 0.143
3 212.1 2.89 2.99 0.152±0.004 0.079
4 247.9 2.89 2.99 0.085±0.001 0.291
5 212.1 1.11 5.23 0.915±0.003 0.067
6 247.9 1.11 5.23 0.120±0.008 0.162
7 212.1 2.89 5.23 0.230±0.008 0.080
8 247.9 2.89 5.23 0.098±0.003 0.305
9 200.0 2.00 4.11 0.650±0.002 0.081
10 260.0 2.00 4.11 0.092±0.007 0.375
11 230.0 0.50 4.11 0.554±0.001 0.076
12 230.0 3.50 4.11 0.101±0.003 0.180
13 230.0 2.00 2.20 0.118±0.003 0.139
14 230.0 2.00 6.00 0.170±0.003 0.190
15 230.0 2.00 4.11 0.109±0.004 0.166
16 230.0 2.00 4.11 0.110±0.002 0.164
17 230.0 2.00 4.11 0.109±0.004 0.163
a
Values expressed as % of oleic acid; b Calculated by difference

CONCLUSION
It was possible to conclude that for the physical refining of palm oil, the variables temperature and
steam percentage presented the predominant effects. However, for the response final FFA the effect
of the initial oil acidity was also significant, providing evidences of the need for the previous
analysis of the oil before sending it to the commercial refining process.

REFERENCES
[1] Tan, C.-H., Ghazali, H. M., Kuntom, A., Tan, C.-P. and Ariffin, A. A. 2009. Extraction and physicochemical
properties of low free fatty acid crude palm oil, Journal of Food Engineering, 113(1), 645-650. [2] Rossi, M.,
Gianazza, M., Alamprese, C. and Stang, F. 2001. The Effects of Bleaching and Physical Refining on Color and
Minor Components of Palm Oil. Journal of the American Oil Chemistry Society, 78(1), 1051–1055. [3]
Sampaio, K. A., Ceriani, R., Silva, S. M., Taham, T., Meirelles, A. J. A. 2010. Steam Deacidification of Palm
Oil, Food and Bioproducts Processing, doi:10.1016/j.fbp.2010.11.012. [4] BOX, G.E.P. and HUNTER, J.S.
1978. Statistic for Experimenters – An Introduction to Design, Data Analysis, and Model Building. John Wiley
& Sons, New York, USA.

1954
Challenges and solutions of a novel muscle-food processing technology: acid and alkaline

solubilization

Patroklos K. Vareltzis1, Konstantinos G. Adamopoulos1 and Herbert O. Hultin†

1
Faculty of Engineering, Department of Chemical Engineering, Laboratory of Food Process Engineering, Aristotle
University of Thessaloniki, 54124 Thessaloniki, Greece
(p.k.vareltzis@gmail.com; costadam@eng. auth.gr)
†
Department of Food Science, University of Massachusetts at Amherst, USA

INTRODUCTION
Acid and/or alkali solubilization is a recent method developed to separate muscle proteins with good
functional properties. However, exposure of the muscle and its components at low pH values has been
shown to promote lipid oxidation. Therefore, the utilization of fatty fish species is limited. The major
factors involved in lipid oxidation process are the substrate, the pro-oxidants and the anti-oxidants of the
system. A main substrate for lipid oxidation is the membrane phospholipids, because they are more
susceptible to oxidation than the triacylglycerols. Hemoproteins, like hemoglobin and myoglobin, are
believed to be the main pro-oxidants in a muscle food system .This research work aims primarily to study
the structural and physicochemical changes of the fish membranes brought about during acid or alkali
solubilization processes. The effect on lipid oxidation and ways to stabilize the membrane lipids against
oxidation are also studied. Model systems comprising minced cod muscle or cod microsomal suspensions
are used. Cod muscle (Gadus morhua) was chosen as it primarily contains membrane lipids with almost
no triacylglycerols.

MATERIALS & METHODS

Fillets of Atlantic cod (Gadus morhua), and haddock (Melanogrammus aeglefinus) were purchased from
a local fish distributor and transported to the laboratory on ice. Chemicals were purchased from Sigma
Chemical Co., (St. Louis, MO). All reagents were of ACS grade and all solvents were of HPLC grade.
Canola oil was purchased from a local supermarket.
Preparation and quantification of hemolysate
A modified method of Fyhn et al. (1979) was used for preparing hemolysate from cod frames (obtained
after filleting whole cod).
Preparation of model systems: protein isolates from washed cod and microsomal suspensions. Protein
isolates, washed cod model system, press juice and microsomal suspensions were prepared as described
by Vareltzis and Hultin (2008). The protein content of the samples, the total lipid content of the minced
muscle and the membrane suspensions were determined according to the method of Vareltzis and Hultin
(2008). A modified method of Lemon (1975) [5] was used for measuring thiobarbituric acid reactive
substances (TBARS).
Data was analyzed by ANOVA using SAS 9.1 (SAS Institute Inc., Cary, NC, USA). Differences between
treatment means at the 5% level were determined using the Duncan Multiple Range Test.

RESULTS AND DISCUSSION

The results clearly indicated that at a pH around 5.3 virtually all of the membranes (99%) in histidine
buffer had precipitated. Sedimentation behavior of membranes in HEPES buffer followed a similar trend,
with a significant (p<0.05) higher however % of supernatant protein at each pH.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1955
To study the effect of pH and added triacylglycerols on hemoglobin-mediated lipid oxidation, a washed
cod/haddock and canola oil system was used. Canola oil (6%, w/w) was added by blending for 1min.
Hemoglobin (6ȝmol/kg of tissue) was added afterwards and the pH of the sample was adjusted to 5.2 or
3.0. After incubation for 20min the pH was readjusted to 7.2 and the samples were put into Erlenmeyer
flasks and stored on ice for oxidation studies. Even though washed haddock was exposed to pH 5.2 in the
presence of hemoglobin, oxidation was slower compared to the untreated sample, where untreated
hemoglobin was added at pH 7.4. TBARS values were significantly lower for the acid-treated sample
compared to the untreated sample. The addition of 6% (w/w) canola oil before pH treatment resulted in
the presence of higher peroxide amounts initially compared to the samples without canola oil. However,
these peroxides exhibited a very slow increase over time (results not shown). Overall, the presence of
canola oil significantly delayed lipid oxidation, when washed muscle and hemoglobin are together
exposed to pH 5.2.
To further investigate the effect of pH on the structural changes of microsomal suspensions, qualitative
SDS-PAGE analysis of treated microsomes was performed. Microsomal suspensions were exposed to pH
3.0 or 10.8 for 30min and then readjusted to pH 7.4. Then the samples were centrifuged and the collected
sediment was resuspended in histidine buffer (pH 7.4). SDS-PAGE analysis of the pH-treated samples
revealed two polypeptide bands ~45 and 36kDa, which were not present in the centrifuged and
resuspended membranes of the untreated microsomal suspensions.
Investigation of hemoglobin binding to membranes showed that that in the case of untreated microsomal
suspensions the precipitation of hemoglobin increases with increasing protein concentration. On the other
hand the acid-treated membranes bind less hemoglobin at high membrane protein concentration, while at
concentration 0.7mg/mL, acid-treated membranes exhibited the highest degree of hemoglobin binding
compared to the other treatments (Table 1).

Table 1. Hemoglobin binding to acid and alkali treated membranes (n=3, ±SE)
Microsomal suspensions with different
% of hemoglobin precipitation
protein concentrations
Hb 8.7± 0.66
MS + Hb (0.7mg protein/mL) 21.76±3.1
*
MS + Hb (1.4mg/mL) 25.33±1.39
MS + Hb (2.2 mg/mL)* 26.33±1.93
*
Acid MS + Hb (0.7mg/mL) 25.33±3.69
Acid MS + Hb (1.4mg/mL) 20.00±2.49
Acid MS + Hb (2.2 mg/mL) 20.66±2.45
Alkali MS + Hb (0.7 mg/mL) 15.83±1.12
Alkali MS + Hb (1.4 mg/mL) 16.33±0.68
Alkali MS + Hb (2.2 mg/mL) 18.66±1.07
Note: the concentrations in the parenthesis refer to membrane protein concentrations. Hemoglobin was added to all samples at a
concentration of 6ȝm; * denotes significant (p<0.05) higher Hb precipitation

Furthermore, the results indicated that press juice retained its anti-oxidative properties after the pH shifts
and could significantly improve the oxidative stability of the samples, even when press juice and
hemoglobin were treated together at pH 3.0.

1956
 Effect of various proteins on characteristics and synerisis of tzatziki

Athanasios G.Stefanakis1, Efstratios K.Stavrakakis1, Konstantinos G. Adamopoulos1, Patroklos K. Vareltzis1,

Athanasia M. Goula2
1
Department of Chemical Engineering, Faculty of Engineering, Aristotle University of Thessaloniki, Thessaloniki,
Greece (costadam@eng.auth.gr)
2
Department of Food Science and Technology, Faculty of Agricultural, Aristotle University of Thessaloniki,
Thessaloniki, Greece (athgou@agro.auth.gr)

INTRODUCTION
Tzatziki is a well known Hellenic delicatessen consisted mainly of yogurt, pieces of cucumber garlic and
olive oil. Since tzatziki consists mainly of yogurt, it shares the same quality profile and problems of
product failure. Yogurt is the result of acidic fermentation of milk creating a network of coagulated
proteins. It is the result of disulfide bonding between k-casein and denatured whey proteins as well as
casein-casein aggregation [1]. The common problems yogurt-based products face, are related with protein
gel network stability, variations of acidity, microbiological infection during storage and the synerisis
phenomenon.
Addition of dried dairy ingredients is a common practice in yogurt manufacture. The addition of dried
dairy ingredients causes an increase in density of the protein matrix in the gel microstructure and
reduction of synerisis in yogurt [2]. In this research 3 dried dairy types of proteins were tested: sodium
caseinate, whey protein concentrate (WPC) and albumin which is mainly originated from egg white
serum. The primary aim of this study was to examine the effect of the above proteins, incorporated in
yogurt individually at different concentrations, on the pH and synerisis of a tzatziki gel during storage at 4
o
C. The water holding capacity (WHC) of each protein-enriched sample in different ratios was measured.
Solubility of the added proteins was determined, since it plays a crucial role in delaying synerisis in
yogurt.

MATERIALS & METHODS

Preparation of tzatziki
Cucumber was added to strained yogurt (10% fat) in a ratio of 1:4 cucumber to yogurt. The moisture
content of cucumber was measured and the appropriate amount of water was added to yogurt in order to
achieve the same moisture content with tzatziki. Whey powder concentrate (WPC), albumin and caseinate
sodium were individually added to the samples at concentrations of 1% and 5% w/w for each protein. The
proteins were dissolved in water at 25°C before added to the strained yogurt. One extra sample without
added protein was used as the control sample. All samples were stored at 5°C.
Synerisis and water holding capacity
Both synerisis an water holding capacity were measured by a centrifuge method according to a modified
method of Keogh and O’Kennedy (1998)[3]. A 20g sample of tzatziki was centrifuged at 2.500rpm for
10min at 25°C. The whey expelled was removed and weighed. The synerisis was expressed as the
percentage % of the whey relative to the original weight of the sample. The water holding capacity was
expressed as the percentage % of pellet weight relative to the original weight of the sample.
Solubility of tested proteins
The solubility of protein water solutions was measured according to Markwell et al. (1978) [4]. The
absorbance of the samples was measured at 660nm using a HeȜȚos Ȗ spectrophotometer (Thermo

11th International Congress on Engineering and Food - Athens, Greece, 2011 1957
Spectronic). Bovine serum albumin was used to obtain the protein standard curve in the range of 0-100 ȝg
protein/ml. The pH value of protein solutions was also measured

RESULTS & DISCUSSION

Water content
The water content of tzatziki was measured at 83% w/w which is 5% higher than that of plain strained
yogurt. Total moisture content of the different samples was reduced proportionally to the level of added
proteins due to the increase of total solids in the mixture.
pH value and stability
pH values of the protein-enriched samples appeared to be more stable through time compared to the
control. Samples containing 1% of added proteins exhibited a stable pH around 4, slightly higher than the
control. However, samples containing 5% of added proteins stabilized at significantly higher pH values
around 4.5. The significant changes in pH of the control sample might be attributed to the transformation
of lactose to lactic acid at different rates depending on the culture reaction. On the other hand, protein-
enriched samples exhibit a higher pH throughout storage, probably due to the dissolved proteins. The
elevation of pH in tzatziki samples can be partly attributed to the pH of added protein solutions which
was in all cases above 6.7.
Synerisis
Overall, protein-enriched samples displayed less synerisis than
the control sample. Higher total solids could cause an increase in
density and reduce pore size in the protein matrix of the yogurt
gel [3]. As shown in Figure 1 the presence of added proteins
decreased significantly the rate of synerisis until the 14th day of
storage. At samples of 1% added protein, WPC exhibited the
most stable results reaching an 8% difference compared to the
control and dropping at 4% difference on the 25th day.
Samples containing 5% of added proteins seemed to have
Figure 1. Rate of synerisis of tzatziki synerisis rates stabilizing at significantly lower levels than the
simulated samples in different control. At 5% level, albumin, which also had the highest
concentrations of proteins solubility, displayed the best behavior achieving 10 – 13% lower
synerisis followed by casein reaching 7-10% less synerisis effect.
CONCLUSION
The addition of proteins to yogurt in order to avoid or delay synerisis seems to be an effective method to
improve product stability. However, more work is needed in order to determine the optimum conditions
of pH, temperature and concentration of the added proteins.

REFERENCES
[1] M.R. Damin, M.R. Alcantara, A.P. Nunes, M.N. Oliveira 2009. Effects of milk supplementation with skim milk
powder, whey protein concentrate and sodium caseinate on acidification kinetics, rheological properties and structure
of nonfat stirred yogurt, Food science and Technology, 42, 1744-1750. [2] Jiancai Li & Mingruo Guo 2006. Effects of
Polymerized Whey Proteins on Consistency and Water-holding Properties of Goat’s Milk Yogurt, Journal of Food
Science, 71(1), 34-38. [3] M.K. Keogh & B.T. O’Kennedy 1998. Rheology of stirred yogurts as affected by added milk
fat, protein and hydrocolloids. Journal of Food Science, 63(1), 108-112. [4] Markwell, M.A., Haas, S.M., Bieber, L.L.
and Tolbert,N.E. 1978. A modification of the Lowry procedure to simplify protein determination in membrane and
lipoprotein samples. Anal. Biochem. 87, 206-210

1958
 High-Power Ultrasound-Assisted Pasteurisation Of Honey

Dania Kabbania, b, Francesc Sepulcreb, Jan Wedekinda

a
Innovació i Recerca Industrial i Sostenible Co, Castelldefels, Spain (dkabbani@iris.cat).
b
Departament d'Enginyeria Agroalimentària i Biotecnologia, Escola Superior d'Agricultura, Universitat Politècnica
de Catalunya, Barcelona, Spain.

INTRODUCTION

Crystallization of honey is a natural but undesirable process in the honey industry. Liquid honey is preferred by most
of the consumers and by food companies for ease of handling. Honey is commonly heated for pasteurization and in
[1]
order to liquefy it and inhibit any microbial growth . However, high temperatures treatments may deteriorate the
quality of the honey. A better method compared to expensive and time-consuming heating is desirable for accelerating
liquefaction and decontamination. In the present work, we liquefied crystallized honey harvested directly from Spanish
apiaries using an ultrasound bath, operating at a nominal frequency of 40 kHz in a temperature range of 40-50 °C for a
maximum period of 120 minutes. We performed several microbial tests to study the degree of microbiological
decontamination after an ultrasound treatment, in particular studying the presence of yeasts and moulds. On the other
hand, honey is also known for its antimicrobial activity due to physical (acidity, osmolarity) and chemical (nectar,
pollen) factors as well as the presence of the glucose-oxidase enzyme. Hence, the effect of the ultrasound on the
antimicrobial inhibitory capacity of the honey was also studied against Saccharomyces cerevisiae.

2. MATERIALS & METHODS

2.1 Experimental design

Crystallized rosemary (Rosmarinus officinalis) honey was obtained from a local producer (Viadiu Ltd., Caldes de
Montbuí Spain).
Samples were placed in 35 ml glass test tubes covered with a screw top, wrapped with parafilm and subjected to the
liquefaction in a temperature range of 40-50 °C for 20-120 minutes by:
a) Liquefaction using an ultrasound (US) bath (Powersonic 603, Cobos Precisions. Korea) operating at a nominal
frequency of 40 kHz (input power 200 W).
b) Liquefaction by only heating (HT) using a thermostatic bath (Digiterm 200 P Selecta S.A, Barcelona, Spain) .
The whole experiment was performed 3 times and samples were prepared by 5 replicates.
2.2 Microbiological analysis and Antimicrobial Activity of honey
Aliquots of one gram of honey samples were placed in sterile Petri dishes and incubated in Nutrient Agar (NA) and
Special Yeast and Mould Medium (SYM) and at 35 ± 1 °C for 5 days. All colonies appearing at the end of incubation
were counted and the results expressed as colony forming units per gram (cfu/g).
To determine the antimicrobial efficacy of honey, Saccharomyces cerevisiae was selected. For the test assays, five
grams of US, HT and raw honey were diluted in 25 ml of distilled water and kept at 20 °C for one hour. A 24 multiwell
plate was used for the screening of the antimicrobial activity. Each well contained 1300 μl of SYM , 500 μl of
inoculum and 200 μl of honey or Buffer solution (control). The plate was measured at intervals of 1h during 24 h at 37
°C.

3. RESULTS & DISCUSSION

3.1 Microbiological analysis

As described in our previous works [3,4] when crystallized honey is sonicated in a nominal frequency of 40 kHz (input
power 200 W) in a temperature range of 40-50 °C it becomes liquid faster than a conventional thermal treatment. This
indicates that honey can be liquefied without comprising its quality by US waves without needing to increase the

11th International Congress on Engineering and Food - Athens, Greece, 2011 1959
 [4]
temperature up to 50 °C or even higher . Raw honey
incubated in NA and SYM mediums showed microbial
contamination. The NA petri dishes presented white
colonies of microorganisms that occupied the totality of
the plates after 24 h. The SYM petri dishes presented a
mean value of 4 dark green colonies of fungi and yeast
after 24 h that also occupied the totality of the plates
within 48 h. Samples were sonicated at 50 °C for 120
minutes and heat-treated in the same conditions but
without sonication. Samples sonicated for 120 minutes in
NA and SYM showed a significant decrease of
Figure 1: Saccharomyces growth curve in presence of
microorganisms of 60 % (45 cfu/g) after 48 and 120 h.
100 microlitres of honey
The US counts in SYM presented an average of just 1
colony per gram while the HT showed a mean value of 4 colonies per gram of treated honey. Moreover, the size of
colonies in the ultrasonicated samples was noticeably smaller. For instance, the diameter of a colony in SYM agar plate
measured 0.8 cm while a colony in the heat treated sample measured 2.5 cm, after 48 h. This indicates that US delays
the microbial growth increasing its stationary phase.
3.2 Antimicrobial Activity of honey
Honey has been reported to have a potent antibacterial activity, effective against a very broad spectrum of species, and
[2]
to have antifungal properties as well . When diluted, honey also shows an antimicrobial capacity due to hydrogen
peroxide, a bacteriostatic agent produced by enzymatic activity in the honey. As previously described [4] US did not
affect the activity of the diastase enzyme. In the present research we aimed at investigating the effect of US on
glucose-oxidase enzyme activity. When diluted honey (100-200 μl) was placed in the presence of Saccharomyces
cerevisiae (103 cfu/ml) and SYM medium and incubated at 37 °C for 24h, a microbial growth was observed.
Nonetheless, it can be seen (Fig.1) that the curve of the sonicated samples showed the lowest development of culture
growth. Different volumes were placed in the multiwell plate (100-200 μl) to observe the efficacy of the honey. 100 μl
of diluted US sample resulted in a decrease in absorbance of 0.1 units in the Saccharomyces culture after 8.6 hours,
which in turn resulted in an inhibition of the culture growth of 50 %. This could be because of the combined work of
the US treatment and the glucose-oxidase activity, which activity is increased once diluted.

CONCLUSIONS

Our results obtained in this research point to a successful application of the ultrasound technology for decontamination
of honey. Ultrasound treatment can be effectively used for thermal processing of honey, as it was observed a
significant decrease and a delayed the microbiological growth of treated honey at 50 °C for 120 minutes. Moreover, the
natural antimicrobial properties of honey were enhanced by the US treatment as an inhibition of culture growth was
observed. Future works will be focused on optimizing the treatment conditions as well as to study the activity of the
glucose-oxidase enzyme after an US treatment.

REFERENCES
[1] Tosi E.A., Ré E, Lucero H. and Bulacio L. 2004. Effect of honey high-temperature short-time heating on
parameters related to quality, crystallisation phenomena and fungal inhibition. LWT - Food Science and Technology,
37, 669-678.
[2] Molan P.C. 1992. The antibacterial activity of honey: 1. The nature of the antibacterial activity. Bee World, 73(1),
5-
[3] Kabbani D, Sepulcre F, Wedekind J. 2011, submitted. Ultrasound-assisted Liquefaction of Honey. Journal of Food
Engineering.
[4] Kabbani D, Wedekind J and. Sepulcre F. 2011, submitted. Honey hidroxymethylfurfural and amylase content
influenced by ultrasound treatment. Journal of Food Engineering.

1960
 Fourier Transform Infrared (FTIR) Spectroscopic Analysis of Biodegradable Gelatin

Films Immersed in Water

Z. A. Nur Hanani, Y. H. Roos & J. P. Kerry*

Food Packaging Group, School of Food & Nutritional Sciences, University College Cork (UCC), Cork,
Ireland (Joe.Kerry@ucc.ie)

INTRODUCTION
Fourier transform infrared (FTIR) spectroscopy is one of the most important tools used to analyze
food packaging films due to its sensitivity, relatively low cost and speed. Its applications are not
limited to analyzing the properties of petrochemical-based plastics, but can also be used for
biodegradable packaging films manufactured from polysaccharides or protein sources. Protein-
based films (manufactured using casein, whey, gelatin etc.) have attracted enormous interest due to
their biodegradability. Furthermore, protein-based films have good barrier properties compared to
synthetic films. Gelatin is a protein that is commercially obtained from skins and bones of cattle and
pigs following slaughter. According to Wang et al. [1], following the screening of numerous food
ingredients for establishment of film forming ability, gelatin was one of the biopolymer materials
assessed which demonstrated desirable film forming properties. Few researchers have investigated
the use of FTIR as an analytical tool for determining biodegradable film properties, and most
specifically for those manufactured from gelatin. Proteins are comprised of amino acids joined
together by amide bonds. The polypeptide and protein repeat units give rise to nine characteristic
infrared (IR) absorption bands, namely; amide A, B, and I-VII [2]. Amide bands represent different
vibrational modes of the peptide bond. The absorption bands of gelatin films in the IR spectra are
situated in the amide band region; Amide-I represents C=O stretching/hydrogen bonding couple
with COO, Amide-II represents bending vibration of N-H groups and stretching vibrations of C-N
groups, Amide-III is related to the vibrations in plane of C-N and N-H groups of bound amide.
Amide-I band is the most sensitive spectral region to the protein secondary structural. The objective
of this study was to investigate the effect of a water immersion process on biodegradable gelatin
films, via the use of FTIR spectra, in order to determine possible interactions that might occur
between gelatin functional groups and water.

MATERIALS & METHODS

Films were manufactured from gelatin, using a 6 and 8% concentration, derived from bovine skin
(Healan Ingredients Ltd, York, UK). Solutions were stirred using a magnetic stirrer hotplate and
heated to 80°C. Films were cast by pouring pre-heated solutions onto level circular teflon-coated
Perspex plates and dried at 50 ± 5% RH and 23 ± 2 °C. Prior to FTIR attenuated total reflectance
(ATR) spectroscopic analysis, gelatin films were immersed in distilled water from 1 minute up to 1
hour in duration. The spectra of gelatin films were recorded using Varian 600-IR Series FTIR
equipped with horizontal attenuated total reflectance (ATR) ZnSe cell at room temperature.

RESULTS & DISCUSSION

The spectra for 6% gelatine films prior to immersion showed that the band was formed by four
individual peaks; situated at amide-A and free water (3282 cm-1), amide-I (1630 cm-1), amide-II

11th International Congress on Engineering and Food - Athens, Greece, 2011 1961
(1547 cm-1) and amide-III (1239 cm-1). The peak situated around 1034 cm-1 might be related to the
possible interactions arising between plasticizer (OH group of glycerol) and film structure [3].
These results were broadly in agreement with previous studies [2,3]. After immersion in water for 1
minute, the amide-A peak, amide-I and amide-II became broader and shifted to a higher
wavenumber. However, the amide-III peak disappeared after the immersion process, thereby
indicating no vibrations in the plane of C-N and N-H groups of bound amide. The amide-A peak,
amide-I and amide-II became increasingly broad when immersion time increased. Additionally, the
peak (1034 cm-1) which related to the interaction between plasticizer and film was not appeared.
Similar spectra for 8% gelatine films to those obtained for 6% gelatin films were noticeable.
However, the wavenumber of Amide-A for 8% gelatin concentration was higher than that for 6%
gelatin, with a lower wavenumber of amide-I and amide-II peaks. After 1 minute immersion, the
amide-A peak showed a broader peak and shifted to a higher wavenumber. The same behaviour
occurred for amide-I and amide-II peaks.
Pure distilled water showed a strong IR absorbance band at 3349 cm-1 (O-H stretching) and around
1637 cm-1 (H-O-H bending). Meanwhile the strong band for distilled water that held the 6% gelatin
films for 1 hour shifted to a lower wavelength, around 3304 cm-1. However, the band for distilled
water that held the 8% gelatin films for 1 hour increased to 3358 cm-1. The peak for amide-I, for
immersed gelatin films have a similar functional group to that of water, thereby making it difficult
to differentiate between absorbances accruing to the presence of water or gelatin films, thereby
leading to banding overlap. However, this problem was overcome by using a subtraction process.
The shift in the amide-A band after immersion might be due to the hydrogen bonding interaction
between polymer molecules in the film. A shift in the secondary structure (amide-I and amide-II)
might also contribute to the creation of ‘hydrogen bonds’ between the molecules in films with
water. Additionally, when protein films like gelatin are immersed in water, the hydrogen attached to
the amide nitrogen could readily exchange for hydrogens attached to water molecules [4]. In terms
of concentration, 8% gelatin films generated strong amide-II peaks compared to 6% gelatin films.
This might be due to the weak bonding of water molecules with those present in 8% gelatin.

CONCLUSION
The FTIR spectroscopy has successfully shown some changes on the properties of the gelatin films
after the samples immersed in water. The changes can be seen after the films immersed in water for
1 minute. The wavenumber of the amide-A, amide-I and amide-II peaks increased when the time
increased.

REFERENCES
[1] Wang, L. Liu, L., Holmes, J., Kerry, J. F. & Kerry, J. P. 2007. Assessment of Film-Forming
Potential and Properties of Protein and Polysaccharide-based Biopolymer Films. International Journal
of Food Science and Technology, 42, 1128-1138.
[2] Kong, J. & Yu, S. 2007. Fourier Transform Infrared Spectroscopic Analysis of Protein Secondary
Structures. Acta Biochimica et Biophysica Sinica, 39(8), 549-559.
[3] Hoque, M. S., Benjakul, S. & Prodpran, T. 2010. Effect of heat treatment of film-forming solution on
the properties of film from cuttlefish (Sepia pharaonis) skin gelatin. Journal of Food Engineering, 96,
66-73.
[4] Gallager, W. 2005. FTIR Analysis of Protein Structure. Manuals Chem 455: Biochemistry Lab,
University of Wisconsin, US.

1962
 Effects of edible chitosan- linseed mucilage coating on quality and shelf life of fresh-cut

strawberry

Laura Eugenia Pérez Cabrera a, Gloria Cristina Díaz Narváez a , Alberto Tecante Coronel b , Chelo
González Martínez c

a
Department of Food Technology, Universidad Autónoma de Aguascalientes, Aguascalientes, México
(leperez@correo.uaa.mx)
b
Department of Food and Biotechnology, Universidad Nacional Autónoma de México, Ciudad de México
c
Department of Food Technology, Universidad Politécnica de Valencia, Valencia, Spain

INTRODUCTION
Application of edible coatings is promising to improve the quality and extend shelf life of
minimally processed produce; several mechanisms are involved in extending the shelf-life of
fruits and vegetables. Strawberries are especially perishable fruits, being susceptible to
mechanical injury, desiccation, decay and physiological disorders during storage [1]. On the
other hand chitosan, is an ideal preservative coating for fresh fruits because of its film-forming
and biochemical properties [2] and its use in food is particularly promising because of its
‘‘biocompatibility’’, non-toxicity and antimicrobial action. Linseed mucilage is capable to
film-forming [3]; the functionality of linseed mucilage is similar to that of gum arabic [4]. The
objective of this study was evaluated the effectiveness of edible chitosan-linseed mucilage
coatings to extend the shelf-life of strawberries minimally processed.

MATERIALS & METHODS

Edible coatings made with brown linseed (Linum usitatissimum) mucilage (ML) polymer (1.5,
1.2 and 1.0% w/v; FA, FB and FC, respectively) in aqueous dispersion, chitosan with a
deacetylation degree of 75% (1% w/v) (Sigma-Aldrich N. C3646), lactic acid 0.5% (Hycel)
and Glycerol 0.6% (Fermot). The pH of the formulations of edible coatings were ~ 4.5-5.2. It
dipped strawberries (cv Albion), stalks, washed and sanitized in the edible coatings and stored
at 10 ° C to facilitate dry 120 min, then were packed in polystyrene bags at 4 ° C. Uncoated
strawberries were used as a blank and coated strawberries without chitosan in the formulation
was control. Were analyzed weight loss, respiration rate (CO2/O2 RR, static method; PBI-
Dansensor), colour (CIEL*,a*,b*; Minolta CR-400) decay rate (lost of visual quality, arbitrary
scale), texture (simple compression plate; Texture Analyser-TA-XT2,) and microbial growth
(plate count of mesophilic bacteria, total coliforms and yeast and moulds) were modelled
according to Gompertz equation during storage (0, 5, 10, 15, 20 days).

RESULTS & DISCUSSION

Weight loss was expressed as percentage loss of the initial total weight (Table 1) all samples
showed a progressive loss of weight during storage, after the fifth day of storage weight losses
for strawberries coated with chitosan-linseed mucilage (FA, FB & FC) were significantly lower
(p<0.05), this behaviour was kept at the end of storage for blank and control 15% and 12%
weight losses, respectively compared to coated fruit.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1963
The respiration pattern at 0 day for coated fruit differed from that of untreated fruit, after 1h of
processed or coated fruits, CO2 production was lower for all coated strawberries than for the
blank and control. The respiratory quotient was in all coated samples cases close to one,
showing that no change was detected in the metabolic pathways of coated strawberries. The
addition of this type of coating is likely to modify the internal atmosphere without causing
anaerobic respiration, since chitosan-linseed mucilage edible are more selectively permeable to
O2 than to CO2 [2]. One of the best parameters to describe the variation of colour is the colour
differences ('E*), as it shows the total change in all parameters L *, a * b *, the changes
during storage were visually perceptible to an end. Uncoated fruits were darkens, skin colour
becomes less chromatic and surface browning develops. Firmness significantly decreased
during storage for all samples, these changes, which can be attributed to tissue senescence and
cell wall breakdown, as well as to the sample water loss, were significantly less marked in
coated strawberries. Under refrigeration, the antimicrobial activity of chitosan seemed to
influence significantly the growth of mesophilic bacteria. Edible coatings chitosan-based
displayed a strong inhibition for all microbial groups; the presence of chitosan on coated
strawberries were significant difference between uncoated or control strawberries.

Table 1. Physicochemical, microbiological and quality parameters of strawberry

% Gompertz parametersa,c
% %
Respiratory Colour Visual
Sample Weight Firmness
a
quotient (RQ) 'E*a a lost A PmaxG O R2
lost lost (N) a
quality
Blank 15r1.5 1.8r0.5 8.2r1.1 55r12 75r5 6.9 3.0 0.45 .995
Control 12r1.0 1.3r0.4 12.3r1.0 52r8 63r2 4.3 7.0 0.39 .998
FA 8r0.8 1.0r0.2 2.1r0.8 6r2 10r1 1.7 2.5 0.18 .995
FB 7r0.5 1.1r0.3 1.5r0.6 6r3 12r2 1.3 2.6 0.21 .997
FC 4r0.5 0.8r0.2 1.8r1.0 8r2 13r1 1.5 3.0 0.15 .989
a
0o20 days, b 0 days, cFor mesophilic bacteria

CONCLUSION
The use of edible chitosan-linseed mucilage applied at strawberry fruits is recommended to
control browning, firmness lost, microbial growth and decay in strawberry fruit in combination
with other methods, i.e. low temperature and suitable packaging.

REFERENCES
[1] Tournas V. H. & Katsoudas E. 2005. Mould and yeast flora in fresh berries, grapes and citrus fruits.
International Journal of Food Microbiology, 105, 11í17.
[2] El Ghaouth A. Ponnampalam R. Castaigne F. & Arul J. 1992. Chitosan coating to extend the storage
life of tomatoes. Horticultural Science, 27, 1016–1018.
[3] Hernández C., Pérez-Cabrera L. E. & González-Martínez C. 2010. Development of Linseed-
Mucilage Edible Coatings and its Application to Extend Fresh-cut Cucumber Shelf-life. In Regalado
C. & García B. E. (Eds.). Innovations in Food Science and Food Biotechnology in Developing
Countries. Inc. AMECA, A. C. Queretaro, Mexico.
[4] Mazza G. & Biliaderis C. G. 1989. Functional properties of flax mucilage. Journal of Food Science,
54, 1302-1305.

1964
 How to apply acrylamide mitigation tools in food technology

Zuzana Ciesarováa, Kristina Kukurovaa, Lucie Markovaa, Jana Sadeckaa

a
VUP Food Research Institute Bratislava, Slovak Republic, e-mail: ciesarova@vup.sk

INTRODUCTION
A high concern about acrylamide has been revealed from the fact that acrylamide belongs to
probably carcinogenic compounds in humans based on the evaluation of the International
Agency for Research on Cancer since 1994. During the last decade acrylamide was confirmed
to be found in many staple foods in a level attracting attention of food safety bodies and
consumers as well. As main sources of acrylamide intake are considered to be, besides coffee
and potato specialities like potato chips and crisps, a wide range of cereal based products
(bread, crisp bread, cookies, crackers, cakes, gingerbreads, breakfast cereals) which
substantially contribute to human exposure of acrylamide. Although cereal products generally
are not known with higher acrylamide content (usually up to 1000 μg/kg), due to their frequent
consumption are considered to be a significant contributor to acrylamide exposure. The reason
arose from the mechanism of acrylamide formation: the simple saccharides, as glucose and
fructose are, act as promoters of acrylamide formation [1].
The incessant effort of responsible food safety bodies as well as research on acrylamide
mitigation in foods has been reflected in summarization of possible tools applicable in food
processing and technology with the aim to reduce acrylamide level in products as low as
reasonably achievable. The document CIAA Acrylamide Toolbox [2] revealed tools based on
next approaches: i) Intervention in raw material selection; ii) Alteration in recipes; iii)
Adjustment of processing conditions; iv) Final preparation before eating. A proposition of tools
is based on the actual knowledge built on the clarification of mechanism and they are chosen
with the following intentions: i) To decrease the level of precursors in raw materials; ii) To
avoid acrylamide formation during processing of foods;
iii) To facilitate acrylamide elimination.

MATERIALS & METHODS

Among possible methods to decrease acrylamide level the selection of raw material according
to the occurrence of precursors and the use of asparaginase before heat processing and/or the
application of inorganic salts seem to be a very efficient way of acrylamide reduction.
Acrylamide and amino acids were determined by LC/MS/MS technique precisely described
previously [3] with calculation on internal standards.

RESULTS & DISCUSSION

Selection of raw materials. The determination of amino acids in raw material or in flours can
predict the acrylamide formation in final products after thermal processing. Differences in
amino acid profile (Asn, Asp, Gln, Glu) between varieties of grains were clearly documented
on the samples of wheat grains, oat grains and wheat flours. Results showed that in the wheat
flour there was the aspartic acid which had the highest portion among determined amino acids.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1965
In the whole grains of wheat and oat the free L-asparagine occupied the highest ratio of free
amino acids. It is important for the next usage of these grains in the process of production of
bakery ware or breakfast cereals.
Application of asparaginase in pastry production. To avoid alteration in expected sensory
properties of final products the application of L-asparaginase enzyme before heat treatment
was used which resulted in a sufficient decrease of acrylamide content. This approach was
successfully used for fried products, baked cakes and biscuits. Appropriate enzyme pre-
treatment of fried cereal products brought “free-acrylamide” products like donuts [4].
Manufactured biscuit and gingerbread production with enzyme pre-treatment resulted in
important elimination of acrylamide in those products which are intended especially for
consumption by children [5].
Application in bread making process. Last but not least, the effort to mitigate acrylamide
was focused on bread making process. Considering the fact that domestic bread preparation is
rising generally and production of bread mix for bread machine are frequently innovated
according to new trends, the incorporation of improving ingredients into the recipes is easily
under control. The calcium chloride was recognized as the salt which suppressed acrylamide
formation up to 10 % and concurrently improved sensorial as well as functional properties of
the final bread loaf.

CONCLUSION
The enzyme application had an unambiguous advantage in no sensorial impact on expected
properties of final products. Calcium chloride addition improved the quality and safety of
bread. Both mentioned approaches are promising ways to depress acrylamide intake from
cereal foods.

ACKNOWLEDGEMENT
This contribution is the result of the project implementation "The Centre of Excellence for
Contaminants and Microorganisms in Foods" funded by the ERDF. This work was also
supported by the APVV contracts No. LPP 0310-09 and VMSP 0089-09 as well as by the
MARD SR under the contract No. 4697/2009-810.

REFERENCES
[1] Ciesarová Z., Kiss E. & Kolek, E. 2006. Study of factors affecting acrylamide levels in model
systems. Czech Journal of Food Science, 24, 133–137.
[2] Toolbox (2009). The CIAA Acrylamide "Toolbox". Confederation of the food and drink industries
of the EU [online]. 2009. 1-41.
[3] Ciesarová Z., Kukurová K., Bednáriková, A. & Morales, J.F. 2009. Effect of heat treatment and
dough formulation on the formation of Maillard reaction products in fine bakery products – benefits
and weak points. Journal of Food and Nutrition Research 48(1), 20-30.
[4] Kukurová K., Morales F.J., Bednáriková A. & Ciesarová Z. 2009. Effect of L-asparaginase on
acrylamide mitigation in a fried-dough pastry model. Molecular Nutrition and Food Research,
53(12), 1532-1539.
[5] Ciesarová Z., Kukurová K., Bednáriková A., Marková L. & Baxa, S. 2010. Influence of food
processing on acrylamide level in gingerbreads and cookies. Aspects of Applied Biology 97(1), 87-
92.

1966
 Coconut water processing using ultrafiltration and pasteurization

L.A.Nakanoa, W.F.Leal Jr.b, D.G.C.Freitasb, L.M.C.Cabralb, E.M.Penhab, A.L.Penteadob, V.M.Mattab

a
Federal Rural University of Rio de Janeiro, Rio de Janeiro, Brazil (lucasassad@yahoo.com.br)
b
Embrapa Food Technology, Rio de Janeiro, Brazil (vmatta@ctaa.embrapa.br)

INTRODUCTION
Green coconut water is a very pleasant drink, usually consumed fresh at production regions,
directed from the fruit. As the water represents just 25% of the fruit weight the discard of
coconut waste is still a problem in the cities [1]. The use of adequate processes for conserving
coconut water makes possible to extend its shelf-life and preserve the fresh characteristics.
When using ultrafiltration membranes, the permeate fraction can be considered as cold
pasteurized as microorganisms are retained by the membrane [2]. In this study ultrafiltration
and pasteurization processes are compared concerning the preservation of coconut water
regarding the fresh product characteristics.

MATERIALS & METHODS

Fresh coconut water was extracted, cold and filtered for removal of suspended solids.
Membrane process was conducted in semi-pilot scale in a plate and frame ultrafiltration unit
composed of 20 kDa flat sheet membranes. Process was carried out in batch mode, at 5 bar and
15oC. The permeate flux was measured along the process. Pasteurization was performed in a
tubular pasteurizer at 96oC for 20 s. Samples of fresh, ultrafiltered and pasteurized coconut
water were collected for determination of physico-chemical and biochemical parameters,
microbiological quality and sensory acceptability.

RESULTS & DISCUSSION

The physico-chemical data are presented in Table 1.

Table 1. Characteristics of fresh, ultrafiltered and pasteurized coconut water

Parameter/Sample Fresh Ultrafiltered Pasteurized
Soluble solids (rBrix) 5.6 5.0 5.2
pH 4.89 4.86 4.89
Acidity (g/100g) 0.06 0.05 0.05
Total phenolics (mg/100g) 3.65 1.09 1.33
Peroxidase activity (U/mL) 0.320 0.002 0.002
Polyphenoloxidase activity (U/mL) 0.649 0.026 0.026

Acidity, pH and soluble solids content of ultrafiltered and pasteurized waters were very close,
being verified a small reduction in soluble solids when comparing to fresh coconut water. The
soluble solids reduction in ultrafiltered water (10%) is in the same range of the sugars

11th International Congress on Engineering and Food - Athens, Greece, 2011 1967
reduction verified by Reddy et. al. [3] when filtered coconut water in a 0.2 μm cellulose nitrate
microfiltration membrane. It can be observed that ultrafiltration and pasteurization were both
effective for reducing the enzyme activity of coconut water although decreasing total
phenolics.
Microbiological analyses have shown that both processes provided safe products with
coliforms at 45oC less than 3 MPN/g, absence of Salmonella spp. in 25 g and counting of
psicrotrophyc bacteria, mold and yeast less than 1.0 x 101 CFU/g.
After consumers’ evaluation, a good acceptance (over to 68%) was observed for all coconut
water samples. Figure 1 shows that the frequency of acceptance scores (scores > 5.0) for
“overall liking” of fresh coconut water was higher than those obtained for the pasteurized and
ultrafiltered coconut water. However, there was a significant difference (p <0.05) in the
acceptability of ultrafiltered water in relation to the other ones. In a study of conservation of
green coconut water by membrane filtration, Magalhães et al. [4] showed that 94% of the
consumers who tasted the ultrafiltered coconut water liked the product. However in the cited
study there was no directly comparison with fresh coconut water.

Figure 1. Frequency (%) of hedonic scores for fresh, pasteurized and ultrafiltered coconut water.

CONCLUSION
The obtained results indicate that both processes may be adequate for processing coconut
water, although ultrafiltration needs to be optimized regarding its effect on sensory
characteristics of the product.

REFERENCES
[1] Aragão W.M. (Org.). 2002. Coco pós-colheita (Frutas do Brasil; 29). Embrapa Informação
Tecnológica, Brasília. Embrapa Tabuleiros Costeiros, Aracaju; Brasil.
[2] Carneiro, L., Sá, I.S., Gomes, F.S., Matta, V.M. & Cabral, L.M.C. 2002. Cold sterilization and
clarification of pineapple juice by microfiltration. Desalination, 5(148), 93-98.
[3] Reddy K.V., DAS, M & DAS, S.K. Non thermal sterilization of green coconut water. 2007. Journal of
Food Quality, 30, 466-480.
[4] Magalhães, M.P., Gomes, F.S., Modesta, R.C.D., Matta, V.M. & Cabral, L.M.C. 2005. Conservação
de água de coco verde por ultrafiltração. Ciência e Tecnologia de Alimentos, 25(1), 72-77.
Acknowledgments: To FINEP for the research funding and L.F.Silva and S.M.Pontes for tests support.

1968
 Progressive freeze-concentration: Improvement and applications

Osato Miyawaki

Department of Food Science, Ishikawa Prefectural University, 1-308 Nonoichi,

Ishikawa 921-8836, Japan (osato@ishikawa-pu.ac.jp)

INTRODUCTION
There are three methods for the concentration of liquid food: evaporation, reverse osmosis, and
freeze concentration. Among these, freeze concentration is known to give the best quality. The
conventional method of freeze concentration is based on the suspension crystallization, in which
many small ice crystals are formed. This system is very complex to require very high initial
investment. On the contrary, the progressive freeze-concentration (PFC) is a method with a single
ice crystal formed on the cooling plate. This method is expected to be much simpler in its system as
compared with the conventional method based on the suspension crystallization.
In the present paper, PFC was applied for the high quality concentration of fruits (pear) flavour
solution, apple juice, tomato juice, and water melon juice. When the osmotic pressure of the sample
to be concentrated was low, a single step PFC was effective with a high concentration yield for
solute components. When the osmotic pressure of the sample was high, some part of the solute was
incorporated into the ice phase to reduce the yield. In this case, however, partial melting of ice was
effective to improve the concentration yield.

MATERIALS & METHODS

A model solution of pear (La France) flavour components, apple juice, tomato juice, and water
melon juice were used as samples to be concentrated. A test apparatus with a vertical cylindrical
sample vessel was used for the small-scale PFC of 100 to 200 mL sample. The sample vessel was
plunged into a cooling bath (-15 oC) at a constant speed to control the ice crystal growth rate. The
sample vessel was equipped with a stirrer to control the mass transfer at the ice-liquid interface. For
the scale-up of PFC, a tubular ice system with 10 L scale was used. In this case, a circulation pump
was used instead of stirring.
Rotary evaporator with reduced pressure operated at 50 oC and reverse osmosis system with a test
vessel (C40-B, Nitto Denko) with a reverse osmosis membrane (NTR-70 SWC, Nitto Denko) were
also used for the concentration of pear flavour components.
The pear flavour solution was extracted with diethylether, which was analyzed chromatographically.
For identification and quantification of the flavour components, GC/MS system (Focus DSQ 2,
Thermo Fisher Scientific) and gas chromatograph (G-3900, Hitachi) were used with a capillary
column (InertCap WAX, GL Science). In the concentration of apple juice, tomato juice, and water
melon juice, the concentration in Brix was measured by a refractometer (APAL-1, Atago).

RESULTS & DISCUSSION

In Fig. 1, the concentration yields were compared for each flavour components among the three
methods. The concentration yields for most flavour components in the evaporation method were less
than 20% and those in the reverse osmosis were less than 60%. In PFC, the concentration yields for

11th International Congress on Engineering and Food - Athens, Greece, 2011 1969
most flavours, except esters, were the highest among the three concentration methods to be 80 to
100%.
In the evaporation, most flavour components were lost in gas phase in the concentration process in
spite of the reduced evaporation temperature down to 50 oC. In the reverse osmosis, the lost
components were detected in the permeate although a tight RO membrane was used. In PFC,
however, the lost components were not detected in the ice phase so that the lost components have
been not incorporated into the ice phase. The relatively lower yields for esters might be related to
the open-air structure of the cylindrical sample vessel with stirring.

Evaporation Reverse osmosis

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Figure 1. Comparison of concentration yield among the three methods for concentration of pear flavour solution.

When the osmotic pressure of the solution was low, high concentration efficiency was obtained with
a high yield by a single step PFC as shown above. With an increase in the osmotic pressure,
however, concentration yield decreases because of the incorporation of solute components into the
ice phase.
Apple juice, tomato juice, and water melon juice were freeze-concentrated by about two-fold by the
tubular ice system. The concentration yield for apple, tomato, and water melon juices were 73.6,
91.2, and 80.3%, respectively. In these cases, the loss components were incorporated into the ice
phase.
The incorporation of solute into the ice phase has been supposed to be the weak point of PFC. To
improve this, the partial melting of ice was applied. When the contaminated ice crystal was partially
melted, the initial fractions of melt ice contained the higher amount of solutes. By recovering these
concentrated fractions, the concentration yield of the process could be improved to a necessary
level. According to this principle, partial melting of ice formed after the PFC of apple juice was
carried out. The concentration of the initial ice melt fractions were high and the concentration yield
could be improved to 90% by recovering the initial 30% of the melt fractions. This shows the
effectiveness of the partial melting of ice to improve the concentration yield in PFC.

CONCLUSION
PFC was proved to be effective for the concentration of solute for the solution with a low osmotic
pressure. In this case, no incorporation of solute components into the ice phase was observed. For
solutions with a high osmotic pressure, however, some part of solute components was incorporated
into the ice phase to reduce the concentration yield. In this case, partial melting of ice was useful to
improve the yield to a necessary level.

1970
Study of color and shrinkage of Physalis peruviana during convective drying by computer

vision

Luis Puentea, Claudia Pintoa, Eric Echegaraya, Eduardo Castroa, Misael Cortésb

a
Universidad de Chile, Food Science and Chemical Technology Department,
Santiago, Chile (lpuente@ciq.uchile.cl)
b
Universidad Nacional de Colombia, Agrícola and Food Engineering Department,
Medellín, Colombia (mcortesro@unal.edu.co)

INTRODUCTION
Physalis peruviana fruit is a juicy berry with ovoid shape, a diameter from 1.25 to 2.50 cm and
a typical weight between 4 to 10 g. The fruit is covered by a calyx, that protect the fruit during
grow and development from insects, birds, diseases and climatic conditions. The fruit is
attributed to different medicinal properties such as antiasthmatic, diuretic, antiseptic and other
effects on human health, more information can be found in Puente et. al., 2011 [1].
One of the most important physical changes that food suffers during drying are the reduction of
its external volume and color. Loss of water and heating cause stresses in the cellular structure
of the food leading to change in shape and decrease in dimension [2]. When water is removed
from food structure, there is disequilibrium between pressures inside and outside the food
matrix structures. This fact generates a contraction of cells belonging to food tissues which
results in shrinkage or collapse of structures. The main objective of this work was to
characterize physically the fruit Physalis peruviana during hot air drying, determining
shrinkage of the sample and its color changes during the process.

MATERIALS & METHODS

Fruits of physalis peruviana L acquired from local market in Santiago-Chile were stored at 5ºC
until drying experiments. A Black box for image acquisition was designed and built according
to sample manipulation and experimental conditions. Inside the box can be found a standard
illumination system (two fluorescent tubes 6500 K), a digital camera Canon ® model Power
Shot 520 and a device to rotate the sample up to 360º. Physalis samples were dried by hot air in
a convective dryer at 65ºC and an air velocity a 1 m/s, pictures of fruit were taken every 15
minutes during the first 4 hours and then every 1 hour until to reach a final moisture of 10%
(w.b.). At the same time moisture content was registered to build a typical drying curve. In this
work a routine in MATLAB ® was developed to process the images and deliver the RGB
values of each image, then these values were converted to L * a * b * by the gamma correction
factor. Volume changes were determined by the technique of solid displacement using poppy
seeds of known density as displacement fluid.

RESULTS & DISCUSSION

Changes in volume and color during drying occurred as a result of drying temperature and
prolonged drying times, in the case of volume (Fig 1, a) can be appreciated, a kind of

11th International Congress on Engineering and Food - Athens, Greece, 2011 1971
anisotropic shrinkage since shrinking is mainly through equatorial axis, while axial axis suffers
almost imperceptible changes., the distribution of seeds inside the fruit determined the final
shape taken by the dried fruit and prevented the development of ideal shrinkage of the sample.
The values of color parameter L* and b* decrease, while the parameter a* increased during
drying as can be seen in Fig 1 b, the fruit becomes darker and less shining, due to the loss of
water and the effect of drying temperature on physalis peruviana structure and chemical
composition.

Figure 1. Changes in Physalis peruviana during drying a) volume, b) color

CONCLUSION
According to the results obtained, it can be established that the Physalis peruviana fruit
undergoes an anisotropic shrinkage during the drying at used temperature, where seeds play an
important role on the final shape of the dried fruit.
The drying process affects negatively the color properties of the fruit.
The experimental device developed was an inexpensive and simple way to study changes in
volume and color of Physalis peruviana fruit during drying.

REFERENCES
[1] Puente L., Pinto-Muñoz C., Castro E., Cortés M., 2011. Physalis peruviana Linnaeus, the multiple
properties of a highly functional fruit: A review. Food Research International,
doi:10.1016/j.foodres.2010.09.034. In Press accepted paper.
[2] Mayor, L., & Sereno, A. M., 2004, Modelling shrinkage during convective drying of food materials: a
review, Journal of Food Engineering, 61, 373-386.

1972
 Optimization of osmotic dehydration process coupled with ohmic heating using granny

smith apples

Sepúlvedaa, A., Sastryd, S., Morenoc, J., Nuñez, H., Almonacida,b, S., and Simpsona,b, R.
a
Departamento de Ingeniería Química y Ambiental; Universidad Técnica Federico Santa
María;Valparaíso, Chile, Ricardo.simpson@usm.cl
b
Centro Regional de Estudios en Alimentos Saludables, Valparaíso, Chile.
c
Departamento de Ingeniería en Alimentos; Universidad del Bío-Bío; Chillán, Chile.
d
Department of Food, Agricultural, and Biological Engineering, Ohio State University, Columbus, USA

INTRODUCTION
Osmotic dehydration is the process of water removal by the immersion of high water-content
cellular solids in a concentrated aqueous solution, during which solids gain takes place
simultaneously [1]. Mass transfer during osmotic dehydration occurs through the semi-permeable
cell membranes by the dominant resistance in mass transfer in biological materials.
Ohmic heating is a thermal process in which heat is internally generated by the passage of an
alternating electrical current (AC) through a body, such as a food system, that serves as an electrical
resistance.
Previous studies have given us the information necessary to establish the processing of fruits at
different voltages, times and temperatures. The various processing conditions employed will
generate different quality or/and energy savings at the end of the process. Therefore, identifying
the optimum processing conditions will be a natural extension of the previous research.

MATERIALS & METHODS

Raw Material
Granny Smith apples were selected and obtained from commercial sources and then refrigerated
at 2 ºC for preservation purposes. Sucrose (commercial sugar) was utilized as the osmotic
solution.
Experimental device
The experimental setup consisted of a stainless steel cylindrical cell, as shown in Figure 1. Each
cylinder acted as an electrode, and the voltage was generated by a variable transformer that
controlled the level of voltage delivered to the cell, thus heating the osmotic solution (45-65 ° Brix)
containing the apple samples.

Figure 1. Experimental device.

RESULTS & DISCUSSION

11th International Congress on Engineering and Food - Athens, Greece, 2011 1973
Analysis of water activity (aw)
Table 1 summarizes the values of water activity and the percentage of reduction from the baseline of
water activity in each experiment. Experiments 12 (40°C, 65°Brix and 130 V) and 4 (50°C, 65°Brix
and 100 V) had the optimum results, whereas the worst results were obtained in experiment 1 (30°C,
45°Brix and 100 V).
Response Surface Methodology
In most RSM problems, one or more polynomials approach the relation between the response and
the n independent variables are used, either a first-order linear model, a linear model with
interactions between variables, or a second-order model when there is curvature in the system. The
model selection was selected according to the CP statistic, the coefficient of determination (R²) and
the adjusted coefficient of determination (R² adjusted).
Table 1. Water activity and water activity loss (%).
EXPERIMENTAL RESULTS: water activity (aw)

EXPERIMENT Time [min] % aw loss

0 30 60 90 t = 90min
1 0,9895 0,9695 0,9617 0,9539 3,60
2 0,9898 0,9761 0,9538 0,9428 4,76
3 0,9894 0,9527 0,9364 0,9216 6,86
4 0,9870 0,9596 0,9176 0,9057 8,24
5 0,9914 0,9545 0,9453 0,9410 5,09
6 0,9909 0,9768 0,9699 0,9378 5,36
7 0,9912 0,9636 0,9468 0,9415 5,02
8 0,9913 0,9506 0,9308 0,9115 8,05
9 0,9896 0,9712 0,9727 0,9555 3,45
10 0,9920 0,9475 0,9238 0,9194 7,32
11 0,9892 0,9723 0,9547 0,9497 4,00
12 0,9887 0,9450 0,9168 0,9045 8,52
13 0,9901 0,9600 0,9517 0,9399 5,07
14 0,9898 0,9552 0,9387 0,9316 5,88
15 0,9896 0,9505 0,9364 0,9285 6,17

CONCLUSION
By using the Response Surface Methodology, effective results were obtained for finding optimal
parameters. The high values for the coefficients of determination (R2) of the models (Yaw, YXt, and
YXss were 0.9754, 0.9690, and 0.9441, respectively) indicated a good fit. In all three cases, the
models justified about 93% of the data variability.
The optimization results obtained were as expected, being located near or within the limits of the
range of research, with the parameter of water activity being the most significant. However, it is
necessary to corroborate the results with organoleptic measurements of the final product.

REFERENCES
[1] Sutar, P., Gupta, D. 2005. Mathematical modeling of mass transfer in osmotic dehydration of onion
slices. Journal of Food Engineering, 78(1):90-97.

1974
 Analysis of the quality attributes of osmotically dehydrated mango

Maida Khana, Lília Ahrnéb and Jorge Oliveirac

a
Universidade Eduardo Mondlane, Maputo,Mozambique (maida.khan@uem.mz)
Lília Ahrné, SIK, Gothenborg, Sweden (Lilia.Ahrne@sik.se)
Jorge Oliveira, University College Cork, Cork, Ireland (jmfcoliveira@gmail.com)

INTRODUCTION
Drying is one of the most widely used methods to preserve fruits. Osmotic dehydration (OD)
has been reported to be useful prior to drying to produce a variety of shelf-stable dried products
with improved quality attributes such as colour, texture and aroma (Heng et al., 1990). Food
preservation is dependent on colour, texture and water activity and those are the quality
parameters that are widely used to evaluate the quality of food. The objective of this work was
to assess the influence of osmotic dehydration and its operating conditions on the quality
factors of mangoes. The parameters measured are related to colour, firmness and water activity.

MATERIALS & METHODS

Two varieties of mango from two different harvest years were used: Haden, 2004 and Haden 2,
2005 variety and Keith variety, 2005. The OD experiments with Haden 1 were done up to 240
minutes and the quality was analysed for the initial and final product. The OD experiments
with Haden and Keith were done up to a moisture content of 65% (g H2O/g product). The
sampling times and the time needed to reach moisture content of about 65% in the OD product
were predicted using the model developed by Khan et al. (2008). The quality of the products
was analysed in terms of colour, firmness and water activity, and the kinetics of quality change
evaluated for all different processing conditions.

RESULTS & DISCUSSION

The quality of fruits and vegetables is strongly dependent on harvest, cultivars, climate, soil
type and ripeness. This results in a wide variability of the product characteristics. The raw
material presented a high variability, being the firmness the most variable parameter.

The firmness of Haden 2 and Keith were very similar and much higher than the firmness of
Haden 1. The results suggested that the Haden 1 mangoes were used in a much more mature
stage compared to the Haden 2 and Keith.

The moisture content values of fresh samples were 82± 1 % for Haden 1, 78±2 % for Haden 2
and 80±2. The soluble solids fraction ranged from 0.14 ±0.02 to 0.17±0.01 being the highest
value for Haden 1, and the water activity was 0.991± 0.003 and 0.989±0.003 for Haden 1 and
Haden 2, respectively, and 0.988±0.003 for Keith. All batches were therefore statistically
similar regarding these parameters. Comparing the three mango batches, Lightness (L), ranged
from 64.2±4.4 to 73.3±3.4. The lowest lightness was for Haden 1. The redness was higher for
Haden 1 than for Haden 2 and Keith, which were more similar. Correlating a and b (redness
and yellowness) and L and b (yellowness and Lightness) showed that the effect of the harvest

11th International Congress on Engineering and Food - Athens, Greece, 2011 1975
year on colour was more important than the variety of mangoes used. It can be concluded that
product homogeneity can be controlled by the state of ripeness, with both varieties being
equally suitable.

None of the quality parameters showed any statistically significant kinetic effect, except water
activity. The final colour quality of the OD mango, Haden 2 and Keith were very similar
compared to Haden1. The yellowness of Haden 1 was high and was even slightly enhanced by
the OD process. The variability was smaller in the OD processed products compared to fresh
products.

Table 1. Average firmness, water activity, composition and colour parameters of the Haden 2 and Keith
OD mangoes
aw Xw Xss L a b F
H2F 0.988±0.003 0.78±0.02 0.14+0.02 73.1±3.6 8.6±5.7 60.1±5.9 5.17±4.47
H2OD 0.973±0.002 0.63±0.02 0.28±0.02 71.0±3.4 10.3±4.9 61.7±6.6 4.08±3.65
KF 0.989±0.003 0.80±0.02 0.14±0.02 69.4±4.2 9.0±2.3 58.1±4.4 5.67±3.87
KOD 0.976±0.002 0.64±0.01 0.23±0.01 66.1±4.0 9.6±2.2 60.5±4.5 4.80±3.31
KF = Keith Fresh; KOD =Keith OD processed to 65% water content

CONCLUSION
The Keith and Haden varieties are very similar both in terms of their colour, texture and water
activity parameters as fresh, and after OD. OD did not affect colour nor texture significantly.
However, OD significantly reduced the spread of texture and water activity, that is, it resulted
in a more homogenous product.

REFERENCES
[1] Bolin, H.R. & Huxsoll, C.C. 1993 Partial drying of cut pears to improve freeze/thaw texture. Journal
of Food Science, 58, 357-360.
[2] Forni, E. Torregiani, D. Battiston, P & Polesello, A. 1986 Research into changes of pectic substances
in apricots and peaches processed by osmotic dehydration. Carbohydrate polymers, 6, 379-393.
[3] Heng, K; Guilbert, S; Cuq, J.L .1990 Osmotic dehydration of papaya - influence of process variables
on the product quality. Sciences des aliments.10, 4, 831-848.[4] Holdsworth, S.D. 1979 Fruits. In:
Priestley R. J. (Eds). Effects of heating in Food stuffs. London: Applied Science Publishers Ltd. pp
255-305.
[4] Khan,MAM, Ahrné,L, Oliveira, JC and Oliveira, FAR. 2008. Prediction of water and soluble solids
concentration during osmotic dehydration of mango, Food and Bioproducts Processing, Volume 86,
Issue 1, March 2008, Pages 7-13.
[5] Torregiani, D., Forni, E., Maestrelli, A. & Quadri, F. 1999. Influence of osmotic dehydatration and
pectic composition of kiwifruit slices. Drying Technology, 17: (7&8), 1387-1397.

1976
Osmotic Dehydration Process Coupled with Ohmic Heating Using Granny Smith Apples

and its Effects on Product Quality

Simpson a,b, R., Fariasa, C., Medinaa, V., Almonacid a,b, S., and Nuñeza, H.
a
Universidad Técnica Federico Santa María, Departamento de Ingeniería Química y Ambiental, P.O.
Box 110-V; Valparaíso, Chile.
b
Centro Regional de Estudios en Alimentos Saludables, Blanco 1623, Valparaíso, Chile.

INTRODUCTION
Osmotic dehydration is a water removal process involving soaking of foods, mostly fruits and
vegetables, in a hypertonic solution such as concentrated sugar syrup. This gives rise to two
major simultaneous counter-current mass transfer fluxes, namely water flow from the product
to the surrounding solution and solute infusion into the product [1], [2], [3]. Ohmic heating is
defined as a process where electric currents are passed through foods to heat them. The
dissipation of electrical energy in the product-mass generates heat. Several studies have shown
that osmotic dehydration process can be accelerated through ohmic heating of food material.
The objective of this study was to evaluate the quality of Granny Smith Apples when subjected
to osmotic dehydration coupled with ohmic heating. Variables included in this study were
temperature (40–60 °C), voltage (100–140 V) and solution concentration (52–68% w/w) at a
processing time of 90 min. The moisture content, soluble solids, water activity and physical
parameters such as color and texture, were determined. Measurements were made in triplicate
at 0, 30, 60 and 90 minutes and correlated through Response Surface Methodology using a
factorial Box-Behnken Design.

MATERIALS & METHODS

Materials
Apples samples (granny smith) were hand-peeled, and cut into cubes of approximately 1 cm3
and were placed in an anti-browning solution. Sucrose, the osmotic agent, was a food grade
commercial granulated cane sugar.
System description
The heater and data acquisition system used consisted of a stainless steel cylindrical cell, with
each cylinder acted as an electrode. Samples were heated using a regulated thermal bath (BS-
21 Jeio Tech, Korea) containing the cell and controlling the temperature during the
experiments. A ratio of apples to sugar solution of 1:5 by weight was used. Apples pieces were
weighed and placed into cylindrical cell containing sugar solutions of varying concentrations
(52–% 68 by weight).
Determination of physicochemical properties
Measurements of water activity, moisture and soluble solids were determined in triplicate at 0,
30, 60 and 90 minutes.
Analysis of quality properties
The International Commission on Illumination (Commission Internationale de IEclairage, CIE)
based L*, a* and b* that represent fruit surface lightness, green and red chromaticity
coordinate, and blue and yellow chromaticity coordinate respectively were measured using a
Colorimeter (Minolta,CR-400 , Japan). Textural properties of sample were measured by using

11th International Congress on Engineering and Food - Athens, Greece, 2011 1977
texture analyzer CT3 (BROOKFIELD, EEUU) with 10 kg load cell and computer software of
texture TexturePro CT. A Kramer cell was used for compression test.

RESULTS & DISCUSSION

Regression equations describing the effect of osmotic dehydration coupled with ohmic heating
variables on the total color difference ('E) and % loss of hardness of granny smith apples are
given in Table 1.
Table 1: Regression equation coefficients.
Regression coefficients Hardness (%) Total color difference ('E)
ȕ0 117,0023 36,51597
ȕ1 19,1711 0,31109
ȕ2 -29,4711 -1,70230
ȕ3 3,8335 0,13617
ȕ4 -0,1606 0,00993
ȕ5 0,0080 0,00434
ȕ6 -0,0144 -0,00005
ȕ7 -0,1075 -0,01273
ȕ8 0,3354 0,00921
ȕ9 -0,0109 -0,00118
R2 0,972 0,660

Relatively high correlation coefficients (R2) were obtained for loss of hardness (%). However,
for the color difference only 66% is explained by the model. The optimal values obtained from
the operating conditions (independent variables) for each objective were as follows:
• The optimal value for % loss of hardness was 0.24 % for the independent variables 47.55 ° C,
53.27 °Brix and 102.84 V.
• The optimal value for color difference was 'E= 1.27 for the independent variables 40 ° C, 68
°Brix and 100 V.

CONCLUSION
By using second order polynomial models fitted in this study, the % hardness loss and total
color difference during drying osmotic dehydration coupled with ohmic heating can be
predicted. RSM and the conventional graphic and desirability functions methods have been
effective in determining the optimum zone within the experimental region. The high value for
the coefficient of determination (R2) of the % model loss of hardness indicated a good fit. In
the case of color difference the correlation was only 66% so in future studies should better
characterize the raw material.

REFERENCES
[1] Azoubel and Murr, 2004 P.M. Azoubel and F.E.X. Murr, Mass transfer kinetics of osmotic dehydration of cherry
tomato, Journal of Food Engineering 61 (2004), pp. 291–295. [2] Lazarides et al., 1995 H.N. Lazarides, E. Katsanidis
and A. Nickolaidis, Mass transfer during osmotic pre-concentration aiming at minimal solid uptake, Journal of Food
Engineering 25 (1995), pp. 151–166. [3] Lewicki and Porzecka-Pawlak, 2005 P.P. Lewicki and R. Porzecka-Pawlak,
Effect of osmotic dewatering on apple tissue structure, Journal of Food Engineering 66 (2005), pp. 43–50.

1978
 Physico-chemical, rheological and sensory properties of shamia date sheets

Khaled Youssef, Adel Shatta, Tamer Moussa-Ayoub, Salah El-Samahy

Faculty Of Agriculture, Suez Canal University, Ismailia, Egypt

ABSTRACT
Shamia –dry variety- date pulp (45 ºBrix) was used to prepare eight different treatments of date
sheets. Citric acid, Na-metabisulfite, eugenia and cinnamon were homogenate with date pulp
and spread on aluminum trays, and then dried at 60 ºC for 24 h. The sheets were rolled in a
polyethylene bags till analyze for physical, chemical, and sensory properties as well as
rheological properties of control sheet either before drying or after rehydration.
Moisture content of the sheets ranged from 14.49 to 17.22%. Adding citric acid, Na-
metabisulfite alone or as a mixture significantly decreased the color index and hydroxyl methyl
furfural content, while increased the lightness value of the resultant sheets. Reducing sugars
significantly increased by adding citric acid, while fiber content slightly decreased. There were
some variations between the rheological properties of the date pulp before and after
dehydration. While, date sheets containing citric acid or a mixture of citric acid with Na-
metabisulfite had the highest organoleptic scores the sheets containing eugenia and cinnamon
had the lowest scores.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1979
1980
 Colour stability of spinach leaves during freeze processing steps

Khaled Yousef, Adel Shatta, Abdullah Al-Sanabani

Faculty Of Agriculture, Suez Canal University, Ismailia, Egypt

ABSTRACT
Chlorophylls content and Hunter color readings (L*, a*, b*) of spinach (Spinacia oleracea)
leaves after washing, blanching and freezing processes were measured. Chlorophyll a content
not significantly decreased during processing steps, but chlorophyll b content significantly
decreased. Hunter color parameter (-a*) values significantly decreased during process. The
correlation between the contents of chlorophylls a and b and Hunter color readings during
processing was linear. The best correlation was between chlorophyll a content and –a* values
(r2= 1.00).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1981
1982
 Study on Combined Hot-air and Microwave Vacuum Drying for Scallion

Yifan Li, Shujun Li*, Bingnan Yang, Qinghua Han, Jiwei Ma, Donglin Zhao

Chinese Academy of Agricultural Mechanization SciencesˈBeijing 100083, China

INTRODUCTION
Scallion is a delicious and nutritious vegetable as raw material or seasoning. Microwave
vacuum drying combines the advantages of very rapid and low-temperature drying, and obtains
dried products of high quality. [1-3] The aims of this study was to investigate the effects of
different drying conditions and drying methods on its drying time and sensory quality
including color, superficial aspect and shape of dried scallion.

MATERIALS & METHODS

Materials
Fresh scallion was purchased from local market, and the processing scallion should be fresh, no
rotten and no bad. The experimental microwave vacuum drying apparatus was previously
developed by the authors and was described previously. [4]
Methods
Fresh scallion was sorted, peeled root and skin, washed, and cut segments of certain length in
the range of 5~25mm. Then scallion segments was dried by four drying methods including
hot-air drying, microwave vacuum drying, combined hot-air and microwave drying, combined
hot-air and microwave vacuum drying, respectively.
The hot-air pre-drying is the same processing in these two combined drying methods at 60
for between 2 and 4 h to reach the required moisture content of about 35%.
A batch of the scallion segments was dried to final moisture content less than 10% at different
four drying methods and various drying conditions. The moisture content was measured
according to the vacuum oven method. [5]

RESULTS & DISCUSSION

Effect of Cutting Length on Dried Scallion Characteristics
Drying effects of scallion for different cutting length( 5, 10, 15, 20 and 25 mm)using
Microwave vacuum drying was studied. It was observed that the shorter cutting length led to
the shorter drying times and the higher sensory quality.Under the drying conditions of
microwave power of 0.65 kW and vacuum level of 0.085MPa, the scallion of cutting length
5mm was the shortest drying time of 18min and the highest sensory quality. When the scallion
cutting length increased to 25mm, the drying time rose to 35min, and furthermore, the sensory
quality became lower so that scallion segments was severe scorch, part of scallion core was
wetting, majority of scallion was not drying and sticking.
Effect of Microwave Power on Dried Scallion Characteristics
The microwave vacuum drying times were 18, 15, and 13 min at the microwave power of 0.65,
1.30, and 1.95 kW, respectively at 5mm scallion cutting length and 0.085MPa vacuum level. It

11th International Congress on Engineering and Food - Athens, Greece, 2011 1983
was obvious that the higher levels of microwave power resulted in little shorter drying
times.While microwave power increasing, the drying speed was so rapid that ultimate water
content of scallion was more difficult to control, the edges or sharp corners of scallion was
easy to overheat, and some core appeared yellow, leading to the lower scallion sensory quality.
[6,7]

Effect of Different Drying Methods on Dried Scallion Characteristics

Drying time of four different drying methods including hot-air drying, microwave vacuum
drying, combined hot-air and microwave drying, combined hot-air and microwave vacuum
drying, respectively are 4.5h,0.6h,2.9h,2.8h at 5mm cutting length, 0.65 kW microwave power
and 0.085MPa vacuum level. The total drying time of combined hot-air and microwave
vacuum drying decreased 1.7h, about 37.8% than the hot-air drying, and increased 2.2h, 3.67
times than microwave vacuum drying. [8]
Sensory quality of hot-air drying was the best. The sensory quality of combined hot-air and
microwave vacuum drying was little worse than hot-air drying, but better than combined
hot-air and microwave drying. The sensory quality of microwave vacuum drying was lowest in
the four drying methods.

CONCLUSION
At the scallion cutting length of 5mm, it was obtained that the shortest microwave vacuum
drying time of 18min and the highest sensory quality. The higher microwave power resulted in
little shorter drying times and obvious lower scallion quality using microwave vacuum drying.
In the 4 different drying methods, the total drying time of combined hot-air and microwave
vacuum drying was lowest under little decrease of sensory quality. So combined hot-air and
microwave vacuum drying is suitable advanced drying technology for scallion drying process.

REFERENCES
[1] Liu Yuhuan, Yang Dejiang, Qin Liangshen. 2006. Research on the freeze-drying processing and the
curve of the onion. Food Research and Development, 27(7), 121-122, 127.
[2] Cui Qingliang, He Baofeng. 2009. Experimental study on Chinese onion vacuum freeze-drying
process. Packing and Food Machinery, 27(5), 46-49.
[3] Han Qinghua, Yin Lijun, Li Shujun, et al. 2010.Optimization of process parameters for microwave
vacuum drying of apple slices using response surface method. Drying Technology, 28(4), 523-532.
[4] Han Qinghua, Li Shujun, Ma Jiwei, et al. 2006. Microwave vacuum drying and puffing characteristics
of apple chips. Transactions of the Chinese Society for Agricultural Machinery, 37 (8), 155-158,167.
[5] Zheng, S.X.; Li, Y.Z.; Luo, et al. 2004. Effect of microwave on the dehydration feature of apple
crispy chips. Journal of South China Agricultural University, 25(3), 109-111.
[6] Zhang Guochen, Mao Zhihuai, et al. 2005. Combined vacuum-microwave and hot-air drying of
scallop. Transactions of the Chinese Society for Agricultural Engineering, 21(6), 144-147.
[7] Giri, S.K.; Prasad, S. 2005. Drying kinetics and rehydration characteristics of microwave-vacuum and
convective hot-air dried mushrooms. Journal of Food Engineering, 78, 512–521.
[8] Wu Haihua, Han Qinghua, Yang Bingnan, et al. 2010. Experiment on combining hot air and
microwave vacuum to dry lycium. Transactions of the Chinese Society for Agricultural Machinery, 41
(supp.), 178-181,202.

1984
 Experimental Study of Vacuum Discharge in Microwave Freeze-drying Process

Youfu Cao, Shujun Li*, Bingnan Yang, Fengmin Zhao, Dan Su, Qingliang Zhao

Chinese Academy of Agricultural Mechanization Sciences, Beijing, China (sxllc@sohu.com)

*Corresponding Author: Chinese Academy of Agricultural Mechanization Sciences, Beijing, China
(lisj@caams.org.cn)
INTRODUCTION
Microwave Freeze-drying technology (MFD) is one of the new promising techniques in recent
years, which has been used successfully as a heat source in the food industry. There are many
advantages of MFD, compared with the traditional freeze drying (FD) technology, which can
greatly reduce the drying time, energy consumption and capital cost [1, 2, 3]. However, there
was little research on how many influence factors in microwave discharge and their interaction.
Through the observation, it is a relatively complex subject that microwave discharge depends
on several factors in microwave freeze-drying process. In this work, the relationship between
microwave critical discharge power, the weight of feedstock, vacuum pressure and energy
feed-in area was mainly studied.

MATERIALS & METHODS

Fresh winter-dates with proper in appearance, same maturity, no mildew, big size and small
kernel were cleaned and cut into 8-mm-thick slices with a hand-operated slicer. Then, the
samples with different weights in trays were frozen at -25ć for at least 8 h until all water in
the samples were frozen into ice.
Discharge experiments were carried out on a microwave freeze dryer equipment (LG-0.2,
Chinese Academy of Agricultural Mechanization Sciences, China). Circumstance temperature
was set at -5ć.The variety of weight of feedstock could be neglected as the experience cycle is
quite short (20 min).

RESULTS & DISCUSSION

Effect of the weight of feedstock, vacuum pressure and feed-in area on critical discharge
microwave power
The weight of feedstock, vacuum pressure and feed-in area were chosen as independent
variables to study the law of microwave discharge power. the three groups of feed-in area were
chosen by 1433 cm2, 350 cm2 and 37 cm2. The samples with 78.8ˁof moisture contents were
divided into seven groups according to their original weight (100, 120, 140, 160, 180, 200, 220
g). Each group was frozen at -25ć for at least 5 h, and each group experiment was run in 38±3
times. By analysis of distribution surface method and regression model, there was negative
correlativity between critical discharge power and pressure as well as weight of feedstock and
feed-in area. From the discharge photograph, the larger feed-in area used during MFD
processing results in a lower critical discharge power so that the discharge energy decreased.
The results indicated that the degrees of three factors affecting the critical discharge power:
feed-in areaηvacuum pressureη weight of feedstock.
Regression model fitting
On account of large amount of critical discharge power (P) data, all results were statistically
analyzed with SAS System to have regression analysis [4]Results of analysis of variance and

11th International Congress on Engineering and Food - Athens, Greece, 2011 1985
t test not shown)The fitted results of predictive model based on weight of sample(M) and
vacuum pressure (N)values( the feed-in area was 1433 cm2 , 350 cm2 and 37 cm2 respectively)
show that all the models were acceptable for high correlation coefficient.The regression
equation, which prediction of the law of microwave discharge in three groups with change of
vacuum pressure and weight of sample, were presented in the following:
P1=-0.0202112M2 +0.0011314MN+7.9732251M+ 0.0983634N-495.9521297 Eq. (1)
P2=161.5202717+0.0116532M2+0.0000436N2㸫2.2142811M+0.0776315N Eq. (2)
P3=149.308+0.009M2+0.00009MN- 1.911M Eq. (3)
Confirmatory studies
The ten groups with different value of weight of feedstock (M) and vacuum pressure were
randomly selected for testing regression models.


 0HDVXUHG9DOXH
0LFURZDYHSRZHUYDOXH

 
 3UHGLFWLYH9DOXH


0HDVXUHG9DOXH
 
 3UHGLFWLYH9DOXH
 
0HDVXUHG9DOXH


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([SHULPHQW1R

Figure 1. Comparison curves of predictive values and testing values

Figure 1 shows that Approximation of models provided better description of the data resulting
in a more desirable prediction of the critical discharge power.

CONCLUSION
In summary, the data from the present study indicate that there was negative correlativity
between critical discharge power and pressure as well as weight of feedstock and feed-in area
Using the SAS software, the regression models based on the ANOVA and significance test
were established. And the tests used in this study show that the predictive value in the
regression model is closer to the measured values and the regression model proved to be an
excellent adaptation. By analysis of distribution surface method and regression model indicate
that The results indicated that the degrees of three factors affecting the critical discharge power:
feed-in areaηvacuum pressureη weight of feedstock. Subsequently, this result can provide
the theoretical basis for the design of microwave freeze-drying equipment.

REFERENCES
[1] Wang Zhaohui.1996. Heat and Mass Transfer During Microwave Freeze-Drying. Southeast
University, Nanjing, China. (in Chinese)
[2] Copson D.A.. 1975. Microwave Heating, AVI Publishing Co. Inc.,New York.
[3] Qian Hongsen.1985.Technique and Application of Microwave Heating. Inc.Science and Technology
Press, Haerbin, Heilongjiang. (in Chinese)
[4] Ren Lou-quan.Optimum design and analysis of experiments(Second Edition). Beijing, Higher
Education Press, 2003.˄in Chinese˅

1986
 Ultrasound application as pre-treatment for drying of fruits

Fabiano A. N. Fernandesa and Sueli Rodriguesb

a
Universidade Federal do Ceará, Departamento de Engenharia Química, Campus do Pici, Bloco 709,
60455-760 Fortaleza – CE, BRAZIL. Email: fabiano@ufc.br
b
Universidade Federal do Ceará, Departamento de Tecnologia de Alimentos, Campus do Pici, Bloco
858, 60455-760 Fortaleza – CE, BRAZIL. Email: sueli@ufc.br

INTRODUCTION
Drying is the most common method of food preservation. Air-drying is generally carried out at
mild temperatures (between 40ºC to 70ºC) to reduce the degradation of the fruit. The process
takes from 8 to 24 hours depending on the fruit that is being processed, on its initial moisture
content, on the desired final moisture content and on the temperature used in the process.
Temperature is maintained by direct heating of the air passing through the samples making the
air-drying process heat intensive and expensive. A pre-treatment can be employed to reduce
air-drying time.
One of pre-treatment that can be used to reduce air-drying time is ultrasound application.
Power ultrasound can produce chemical, mechanical or physical changes on the processes or
products where it is applied. When low frequency power ultrasound is applied, ultrasonic
waves travel through the solid medium causing a rapid series of alternative compressions and
expansions, in a similar way to a sponge when it is squeezed and released repeatedly (sponge
effect). The mechanical and physical effects of acoustic waves can be used to enhance many
processes where mass transfer takes place, drying included. The forces involved by the sponge
effect caused by ultrasonic waves can create microscopic channels that may ease moisture
removal. These microscopic channels can be used by water molecules as a preferential pathway
to diffuse toward the surface of the fruit increasing its effective water diffusivity.
In this paper, an overview of the effects of ultrasound application in the dehydration of eight
fruits (banana, genipap, malay apple, melon, papaya, pineapple, pinha and sapota) is presented.

MATERIALS & METHODS

The ultrasound pre-treatment procedure consists in the immersion of the fruit in water to which
ultrasound is applied. Fruit samples were cut to obtain cubes, cylinders or triangular shaped
slices, depending on the fruit that was studied. Moisture content was determined by heating in
a drying oven at 105oC for 48h, according to AOAC method. The initial soluble solids content
of the fruit (ºBrix) was determined by refractometry. The samples were immersed in distilled
water and were subjected to ultrasonic waves during a period of 10 to 45 minutes. The process
was carried out placing the samples in an ultrasonic bath. Ultrasound should be applied for at
least 10 minutes, because the effect of ultrasound showed to be insignificant at lower times.
The water to fruit ratio was maintained between 3:1 and 4:1 (weight basis). The pre-treatment
was carried out under ambient temperature. The frequency of the ultrasonic waves was 25 kHz
and the intensity was 4000 W/m2.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1987
RESULTS & DISCUSSION
Each fruit presented a different behavior when ultrasound was applied. Melons and pineapples
lost water to the liquid medium during the pre-treatment. The concentration gradient of soluble
solids between the fruit and the liquid medium should result in the gain of water by the fruit
during the pre-treatment. Most fruit presented such behavior, except for melons and
pineapples. These latter fruits present high moisture content (90% for melons and 83% for
pineapples) and were significantly affected by the sponge effect of ultrasonic waves.
Sugar gain was negative for all fruits indicating that the fruit lost soluble solids to the liquid
medium. This result was expected because of the concentration gradient of soluble solids
between the fruit and the liquid medium that favors the mass transfer of soluble solids from the
fruit to the liquid medium. The loss of soluble solids for each fruit was very different, ranging
from 1.5% to 52.9%. Fruits with high initial moisture content lost more soluble solids to the
liquid medium than fruits with low initial moisture content. Fruits with high initial moisture
content may ease the diffusion of soluble solids toward the liquid medium. The exception was
pinha that rapidly lost soluble solids to the liquid medium because of the low cohesion between
the cells in its tissue structure.
The loss of soluble solids was also influence by the effects of ultrasound on the tissue structure
of the fruit. Ultrasonic waves cause a rapid series of alternative compressions and expansions,
in a similar way to a sponge when it is squeezed and released repeatedly (sponge effect). The
sponge effect results in the creation of microscopic channels in the fruit tissue. The tissue
structure of melons was very sensitive to ultrasound application and several microscopic
channels appeared in the tissue structure. Short microscopic channels were formed in papayas.
Few microscopic channels were formed in pineapples and strawberries, but they were long,
increasing the overall mass transfer. Consequently, the sugar loss in pineapples and
strawberries were high (18.9% and 9.5% respectively).
The effect of the ultrasonic pre-treatment was mainly observed during the air-drying stage
where a significant increase in effective water diffusivity was observed. Effective water
diffusivity was affected by ultrasound application in some fruits. The ultrasonic pre-treatment
had a positive effect on drying because the increase in the effective water diffusivity resulted in
shorter air-drying time if compared to the fresh fruit with no pre-treatment.

CONCLUSION
Fruits pre-treated with ultrasonic waves have presented significant loss of sugars when the
process was carried out using distilled water as the liquid medium. This process can be applied
to produce dried fruits with low sugar content, which might be used in the production of
foodstuffs with reduced calories.
The application of the ultrasound pre-treatment increased the water diffusivity of the fruit, in
most cases. This phenomenon may occur because of the formation of micro-channels during
the application of ultrasound. The increase in the effective water diffusivity at the air-drying
stage makes the use of ultrasound an interesting technique that can be used complementary to
classical air-drying.

ACKNOWLEDGEMENT
The authors thank the financial support of the Brazilian funding institute CNPq.

1988
 A Basic Investigation On Instant Coffee Production By Vacuum Belt Drying.

Katrin Burmestera, Arne Pietschb, Rudolf Eggersa

a
University of Technology Hamburg-Harburg, Germany (Katrin.Burmester@tuhh.de)
b
Eurotechnica GmbH, Hamburg, Germany (arne.pietsch@eurotechnica.de)

INTRODUCTION
Coffee is the internationally second most important merchandise with a worldwide production
of about 8.26 mio tons per year. Instant coffee is an invention of the early twentieth century
and nowadays it plays an important role besides other preparation techniques due to easy
handling and longer durability. State of the art processes for instant coffee production are spray
drying and freeze drying. Freeze drying leads to the best product quality in terms of aroma
recovery but requires a huge amount of investment and operating costs. Spray drying stands
out due to high production capacities at low energy costs but it results in a higher thermal
impact on the product, which may lead to lower product qualities.
Vacuum belt drying is a well-established process in food technology but it has not been applied
yet to the production of soluble coffee. The application of vacuum belt drying could combine
an economically method with a high product quality. [2]

MATERIALS & METHODS

Drying experiments were performed with coffee extract on a vacuum belt dryer in laboratory
scale (Merk process GmbH, 79725 Laufenburg, Germany). Process parameters were changed,
analyzing the product temperature. After each drying experiment samples were taken and the
dry matter content was investigated gravimetrically. Flavour, structure, density and porosity of
the produced instant coffee samples were measured.
The effective diffusion coefficient was evaluated by drying a sample of coffee extract within a
defined surrounding according to the thin-film measurement technique [4].
The sorption behaviour was analysed by a static measurement using eight desiccators
containing different super-saturated salt solutions assuring a fixed water activity within the air.
The samples were stored within the desiccator until its mass was no longer changing.

RESULTS & DISCUSSION

Within this study it was found, that the drying under low pressures (1 – 15 mbar) leads to a
highly porous product cake with a very low thermal conductivity slowing down the further
drying process. Furthermore, the highly porous structure leads to an undesirable appearance of
the product. The analysis of the product’s flavour could directly be related to the product
temperature. The lower the temperature was maintained, the better the flavour retention within
the product. The pressure only influences the flavour retention indirectly by affecting the heat
and mass transfer properties and thereby the product’s temperature. The bulk density is in the
same order for all products from the three different processes. Within this investigation it could
be shown, that the process parameters of the vacuum belt drying process have only a minor
influence on the particle density. The vacuum belt dried sample has the highest real density of
the particles, even higher than the freeze dried sample. The high real density values for the
vacuum belt dried particles, which are in the same order as the real density of a finely ground

11th International Congress on Engineering and Food - Athens, Greece, 2011 1989
powder, indicates that during the drying process only a macroscopic bubble structure is formed
within the porous product cake and no vapour is included within the macroscopic lamella
structure. The SEM micrographs affirm this assumption. Furthermore, it can be seen, that the
particles have a big surface area compared to the spray dried or freeze dried particles. The
combination of high density and big surface area lead to extraordinary good dissolution
properties of the product, which would even allow a cold dissolution of the product.
The recorded thin-film drying curves were analysed using the analytical solution according to
Crank. As proposed by Räderer [4] the thickness of the plane sheet is to be considered as
double sized taking into account that the mass transfer only takes place at one surface. The
effective diffusion coefficient was determined at 50 °C for various water activities of the
surrounding. The temperature dependency of the effective diffusion coefficient is currently
under investigation.
For the sorption behaviour of instant coffee at 40 and 50 °C no significant difference between
the different instant coffees was found. Furthermore, no influence of the product temperature
on the sorption behaviour was detected. A GAB isotherm was determined. The sorption
behaviour of instant coffee was compared to that of green [1] and roasted coffee [3].

CONCLUSION
A series of drying experiments in a lab-scale dryer was performed varying the process
parameters optimising the product’s properties. By minimising the thermal impact on the
product good flavour retention could be achieved. By density and structure analysis of the
product it could be proven that instant coffee produced within a vacuum belt dryer, fulfils the
present requirements, although first impression is not the usual in the market today. Especially
in terms of dissolution properties the product shows advantages compared to standard freeze or
spray dried products. Additionally, the mass transfer throughout the drying process was
analysed, determining the effective diffusion coefficient by means of a thin film measurement
technique. The sorption behaviour of instant coffee was systematically investigated and could
be described using a GAB sorption isotherm. No influence of the production process or the
product temperature on the sorption behaviour was detected.

ACKNOWLEDGEMENTS
This research project was supported by the German Federal Ministry of Economics and
Technology (via AIF) due to a resolution of the German Bundestag. Project BWT 08-001 /
206032.

REFERENCES
[1] Burmester K. & Eggers R. 2010. Heat and mass transfer during the coffee drying process. Journal of
food engineering, 99 (4), 430-436.
[2] Burmester K., Fehr H., Eggers R. 2011. A comprehensive study on thermophysical properties for an
innovative coffee drying process. Submitted to the international journal of drying technology for
publication.
[3] Rahmann S. 1995. Food Properties Handbook. Water activity and sorption properties of food. CRC
Press
[4] Räderer M. 2001. Drying of viscous, shrinking products: modelling and experimental validation.
Dissertation thesis, TU München, VDI Verlag

1990
 Shrinkage of papaya (Carica papaya L.) during convective drying: Influence of glass

transition phenomenon

Louise Emy Kurozawaa,c, Miriam Dupas Hubingerb, Kil Jin Parkc

a
Department of Food Technology, Rural Federal University of Rio de Janeiro, Rio de Janeiro, Brazil
(louisek98@gmail.com)
b
School of Food Engineering, University of Campinas, Campinas, Brazil (mhub@fea.unicamp.br)
c
School of Agricultural Engineering, University of Campinas, Campinas, Brazil (kil@feagri.unicamp.br)

INTRODUCTION
An important change in the physical state of the product during drying is shrinkage, which may
be highly influenced by glass transition temperature (Tg) during drying. Fruit samples, due to
its high moisture content, are rubbery state at the start of the drying and could be remain
likewise as process progresses or suffer phase transition for glassy state. At rubbery state,
shrinkage almost entirely compensates for moisture loss. At glassy state, the product would
become rigid and volume reduction is greatly restricted [1, 2]. The aim of the work was to
study the shrinkage during convective drying of papaya (Carica papaya L.) cubes and relate its
extension with glass transition.

MATERIALS & METHODS

A convective tray dryer was used in the experiments (carried out at air temperatures of 40°C
and 70°C and air velocity of 1.0m/s). The sample (papaya cube, 30mm × 30mm × 30mm) was
weighed at regular intervals of time until constant weight. The shrinkage kinetics was
accomplished by monitoring of apparent volume during drying. Samples were photographed
using a camera at regular intervals of time. Their lateral areas and length were measured using
the ImageJ® software. Thermocouples were used to analyze the evolution of temperature into
the product (Tp). The values of Tg of papaya as a function of moisture content were based on
another work [3].

RESULTS & DISCUSSION

The shrinkage phenomenon can be related to the glass transition temperature. Figures 1(a) and
1(b) show the evolution of the Tg, Tp and shrinkage (V/V0) throughout drying process at air
temperature of 40°C and 70°C, respectively. Analyzing Figure 1(a), Tp was higher than the Tg
up to moisture content of 0.21 g water/g, showing that the material was in the rubbery state,
which is characterized by high molecular mobility of solid matrix. At low moisture content, Tg
increased, allowing the material to pass from rubbery to glassy state: the dried material became
rigid, decreasing significantly shrinkage extension. At 70°C, Tp was above the Tg throughout
the process (Figure 1(b)), showing that the material did not undergo phase change during
drying and remained on the rubbery state. The sample suffered shrinkage during the whole
process, until the final moisture content, indicating molecular mobility of the solid matrix.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1991
 (a)

(b)

Figure 1. Relationship between Tg, Tp and (V/V0) at: (a) 40°C and (b) 70°C.

CONCLUSION
This work demonstrated the influence of glass transition on shrinkage of papaya cubes during
drying. At 70°C, the experiment was carried out above the Tg, resulting in higher extent of
shrinkage, due to the mobility of the solid matrix. At 40°C, the samples, suffered phase
transition for glassy state, become rigid and volume reduction was restricted. The results will
be valuable to select adequate drying conditions in order to obtaining dried food with quality.

REFERENCES
[1] Mayor, L. & Sereno, A.M. 2004. Modelling shrinkage during convective drying of food materials: a
review. Journal of Food Engineering, 61(3), 373-386.
[2] Bhandari, B.R. & Howes, T. (1999). Implication of glass transition for the drying and stability of dried
foods. Journal of Food Engineering, 40(1-2), 71-79.
[3] Kurozawa, L.E., Hubinger, M.D. & Park, K.J. 2010. Glass transition and sorption isotherm for fresh
papaya (Carica papaya L.). 17th International Drying Symposium (IDS 2010), Magdeburg, Germany,
3-6 October, 2010. Proceedings p. 945.

1992
 Influence of sucrose replacement on colour and texture of kiwi jam

*E. Rosa, I. Peinado, A. Heredia and A. Andrés

Institute of Food Engineering for Development, Universidad Politécnica de Valencia

*e-mail: esrobar@doctor.upv.es

INTRODUCTION
Traditional manufacturing jam’s methods require concentration by heat treatments, which
promotes quality changes that affect sensory and nutritional properties, the latter related mainly
to ascorbic acid losses. Osmotic dehydration can be used for water removing and sugar
addition in jam manufacturing, avoiding the heating process since water removal takes place
by osmosis. On the other hand, osmotic process can also be applied by using dry sugar instead
of sugar solutions, similar to dry salting process commonly applied to meat and fish products.
In these processes, a concentrated solution, rich in volatile compounds, vitamins and water
soluble minerals is generated due to the water out-flow and its volume is considerably lower
than the volume used in traditional osmotic dehydration processes. Additionally, undesirable
effects related with sugar consumption, as caries and diabetes could be solved if sucrose could
be replaced by other sugars with low glycemic index as fructose and isomaltulose.
The aim of this work was to analyze the sugar influence on optical and textural properties of
kiwi spread-products (50 Brix) elaborated by using dry osmosis dehydration and to compare
these spread-products with kiwi commercial jams.

MATERIALS & METHODS

Kiwis (Actinidia deliciosa) acquired in a local supermarket were peeled, cut in 1 cm3 cubes and
sink in chloride water to eliminate possible field residues.
Equilibrium Stage: Dry osmotic dehydration (DOD) method was used to achieve an
equilibrium concentration of 50 Brix. In this method the osmotic agent was placed directly on
the fruit like in meat or fish salting process. The ratio fruit:osmotic agent was calculated by
mass balances. The process was carried out 25 ºC. In this stage either sucrose (control) or a
mixture 50:50 of isomaltulose:fructose were used like osmotic agents.
Jellification Stage: The ingredients in the spread formulations were dehydrated kiwi, osmotic
solution, apple pectin (1, 1.5 or 2 %) and potassium sorbate (500 ppm). The proportion of
dehydrated kiwi:osmotic solution was exactly the same proportion reached at the end of
equilibrium stage. Therefore, effluents were not generated.
Analytical Determinations: Moisture content was determined gravimetrically by method
20.103 AOAC (1980). Soluble solids content (Brix) was measured with a refractometer at 20
ºC (ATAGO 3 T). Water activity (aw) was determined with a dew point hygrometer (FA-st lab,
GBX). pH was determined by a pH-meter (SevenEasy, Mettler Toledo). Colour coordinates
were determined in a Minolta spectrophotometer (model CM-3600d) using a 20 mm plastic
cell (illuminant D65-10º standard observer). Texturometer TA/XT/PLUS-Texture Analyser and
the accessory Back extrusion cell with 35 mm ring [1] were used to obtain the textural
parameters.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1993
RESULTS & DISCUSSION
The results showed that elaborated kiwi spreads could be compared with kiwi commercial jams
in optical and textural properties, since similar physicochemical values were obtained.
Colour Analysis
There was not a clear tendency on the colour coordinates (L*, a*, b*) in elaborated spreads
depending either on the osmotic agent or pectin percentage. Similar results were found in other
studies about spread-products such as tomato ones [2] while, in strawberry spreads colour
difference depending on osmotic agent were observed [3]. On the other hand, kiwi spread-
products showed more similar colour coordinates to fresh sample than commercial jam. This
would be explained mainly as a result of cooking process in traditional jam and the storage
time of these products.
Texture Analysis
The results showed that spread-products formulated with sucrose had higher consistency and
cohesiveness values than the same products formulated with the mixture isomaltulose:fructose.
Similar results were found in tomato spread-products [2]. On the other hand, spread-products
with different consistency and cohesiveness characteristics could be obtained depending on
pectin percentage used, being able to obtain similar values to commercial jam.

CONCLUSION
This study confirmed that dry osmotic dehydration could be a useful technique in fruit spread-
products manufacturing with a similar stability and texture to traditional fruit jams but more
similar to fresh sample in colour terms. Furthermore, it can be considered environmental
friendly since the management of sugar solutions and thermal treatments are avoided.
Additionally, as there are not generated effluents the process yield is higher and in this sense,
the product keep the aromatic compounds and vitamins levels of fresh fruit.
On the other hand, results confirmed that the mixture of isomaltulose:fructose is a good
alternative to replace sucrose in this kind of products since, colour difference were not found
between sugars. In this sense, healthy spread-products suitable to diabetic persons could be
elaborated and in addition, this kind of fruit spreads might be employed in the formulation of
other products like yogurt, cookies or cakes.

REFERENCES
[1] Garcia-Martinez, E., Ruiz-Diaz, J, Martinez-Monzó, J., Camacho, M.M., Martinez-Navarrete, N. &
Chiralt, A. (2002). Jam manufacture with osmodehydrated fruit. Food Research International, 35,
301-306.
[2] Rosa, E., Peinado, I., Heredia, A. & Andrés, A. (2010). Influence of some process variables on
optical properties and texture of healthy tomato spreads. In Food Innova 2010. International
Conference on Food Innovation. Book of Abstracts (ISBN: 978-84-693-5010-2) & USB Artículos
(ISBN: 978-84-693-5011-9). Pedro Fito & Fidel Toldrá, eds., Valencia, Spain.
[3] Peinado, I., Rosa, E., Heredia, A. & Andrés, A. (2008). Estudio comparativo del color en fresa
deshidratada osmóticamente con sacarosa, sorbitol e isomaltulosa. In Actas del V Congreso Español
de Ingeniería de Alimentos CESIA 2008. II Congreso Iberoamericano sobre Seguridad Alimentaria.
Cd-Room/Book of Abstracts (ISBN: 978-84-96736-57-3). Mercè Raventós & Jordi Salazar, eds.,
Barcelona, Spain.

1994
 Influence of dry and wet osmotic dehydration on colour and texture of a spread kiwi

product

I. Peinado*, E. Rosa, A. Heredia and A. Andrés

Institute of Food Engineering for Development, Universidad Politécnica de Valencia, Spain

*e-mail: irpeipar@tal.upv.es

INTRODUCTION
Development of new products which present a good organoleptic quality but also offer other advantages
such as the incorporation of healthier ingredients is currently a priority in the food industry. It is also
important that the elaboration method of these products would be as soft as possible to preserve the
maximum characteristics and nutrients of fresh fruit. In this sense, the elaboration of healthy spread fruits
by osmotic dehydration is an excellent option due to the high quality of the products obtained as
compared with other drying methods.
Traditional wet osmotic dehydration has however some disadvantages related with the handling of big
volumes of osmotic solutions. The dry osmotic dehydration might be an alternative since the volume of
solution generated is considerably lower than the volume managed in the wet method and it is more
concentrated in aromatic compounds, soluble vitamins and minerals as it comes from the product itself.
The aim of this study was to compare the two different methods, wet and dry osmotic dehydration, as a
step to produce a 30 Brix spread kiwi product formulated with two different sugars, sucrose and
isomaltulose. Physicochemical parameters as well as colour and texture analyses were performed.

MATERIALS & METHODS

Kiwis (Actinidia deliciosa), acquired in a local supermarket were peeled, cut in 1 cm3 cubes and sink in
chloride water.
Equilibrium stage: two different osmotic dehydration methods were used, Wet Osmotic Dehydration
(WOD), in which the fruit was immersed in a hypertonic osmotic solution (40 Brix) and Dry Osmotic
Dehydration (DOD), in which the osmotic agent was placed directly on the fruit like in meat or fish
salting process. The equilibrium concentration achieved in this stage was 30 Brix. The ratio fruit:sugar or
fruit:solution was calculated by mass balances. Sucrose or Isomaltulose were used as osmotic agents. The
work temperature was 25 ºC.
Product formulation: the ingredients in the spread formulations were dehydrated kiwi, osmotic solution,
apple pectin (1, 1.5, 2 %) as a jelling agent and potassium sorbate (500 ppm) as a preservative. According
to the different proportions of dehydrated kiwi:osmotic solution and depending on the dehydration
methods, three kinds of process were selected.
x WOD: 70 % dehydrated fruit and 30 % osmotic final solution.
x DOD1: 100 % dehydrated fruit and 100 % generated osmotic solution
x DOD2: 70 % dehydrated fruit and 30 % generated osmotic solution.
Analytical determinations: Moisture content was determined gravimetrically by method 20.103 AOAC
(1980). Soluble solids content (Brix) was measured with a refractometer at 20 ºC (ATAGO 3 T). Water
activity (aw) was determined with a dew point hygrometer (FA-st lab, GBX). pH was determined by a pH-
meter (SevenEasy, Mettler Toledo). Colour coordinates were determined in a Minolta spectrophotometer
(model CM-3600d) using a 20 mm plastic cell (illuminant D65 - 10º standard observer). Texturometer

11th International Congress on Engineering and Food - Athens, Greece, 2011 1995
TA/XT/PLUS-Texture Analyser and the accessory Back extrusion cell with 35 mm ring were used to
obtain the textural parameters.
Table 1. Experimental design of the different kiwi spread products.
Dehydration kind Pectin % Dehydration kind Pectin %
SUCROSE ISOMALTULOSE
1 1 10 1
2 WOD 1.5 11 WOD 1.5
3 2 12 2
4 1 13 1
5 DOD 1.5 14 DOD 1.5
6 2 15 2
7 1 16 1
8 DOD 1.5 17 DOD 1.5
9 2 18 2

RESULTS & DISCUSSION

After the formulation of the different kiwi spreads they were storage during 24 hours at room temperature
to allow the gel formation. Then analysis of moisture (xw), soluble solids content (xss), and water activity
(aw), pH as well as colour and texture determination were performed.
The obtained products showed a similar composition in terms of moisture and soluble solids content
which could be expected as the ratio was calculated to reach a final concentration of 30 Brix. The solid
soluble content of the final spread-products was 0.291 (± 0.03) and the moisture of 0.69 (± 0.02). On the
other hand, aw showed values of 0.964 (± 0.006). Finally, pH value for the formulated products was
around 3.37 (± 0.07). Therefore no differences depending on the different sugars, dehydration methods or
pectin percentages were found in terms of physicochemical parameters on the final products. These
results are contrary to those obtained for the same kind of spreads elaborated with strawberry where the
isomaltulose spreads showed a higher aw compared with the sucrose ones.
Colour analysis: All the samples were very similar to the fresh one and there was not a clear tendency in
the colour coordinates (L*, a*, b*) of samples depending on osmotic dehydration method, kind of
osmotic agent or pectin percentage; similar results have been reported in tomato spreads of 50 Brix, while
colour differences have been noticed in other spread-products such as in strawberry ones depending of
osmotic agent. This fact could be related to the high stability of kiwi pigments and lower degree of
interaction within the system components compared to those present in strawberries.
Texture analysis: No differences in texture parameters were found between the different products as a
consequence of the different sugars used in their formulations. On the other hand, spread-products with
different consistency and cohesiveness characteristics could be obtained depending on osmotic
dehydration method and pectin percentage used.

CONCLUSION
This work confirmed the viability of DOD and the use of new sugars (ex, Isomaltulose) as an alternative
to obtain tomato spreads with colour and texture comparable to those elaborated by WOD with sucrose.
The final product obtained by DOD is additionally richer in aromatic compounds, soluble vitamins and
minerals from fresh fruit since effluents are not generated, getting to increase the process yield.
Furthermore, as isomaltulose has less sweetener power than sucrose, the obtained product is closer in
taste to fresh fruit than the product processed with sucrose.

1996
 Quality assessment of dried eggplant using different drying methods: hot air drying,

vacuum freeze drying and atmospheric freeze drying.

J. V. Santacatalina, C. Ozuna, J. A. Cárcel, J. V. García-Pérez* and A. Mulet

Grupo de Análisis y Simulación de Procesos Agroalimentarios (ASPA), Departamento de Tecnología de Alimentos,
Universidad Politécnica de Valencia, Camí de Vera s/n, E46022, Valencia, Spain (jogarpe4@tal.upv.es).

INTRODUCTION
Drying is a classical method to preserve foods, which provides longer shelf life, lighter weight for
transport and smaller space for storage than fresh products. However, drying prompts a loss of quality
properties due to a structural collapse and biochemical changes. Obviously, the quality degradation
depends on the drying technique used. Eggplant is an important vegetable in Asian and Mediterranean
markets and has a very limited shelf life for fresh use. Dehydration constitutes an alternative to provide
higher stability eggplant products. Dried eggplant could be used as a new ingredient in foodstuffs, like
soups and sauces. Most of dried products should be rehydrated before consumption. Therefore, it is
necessary to know the behavior of the product when it is soaked in water. On the other hand, softening
and loss of texture is a problem of dried/rehydrated products, thus texture profile analysis is frequently
used to evaluate the textural properties of rehydrated products. The main objective of the present study
was to evaluate the effect of different drying techniques on the quality of dried eggplant.

MATERIALS & METHODS

Hot air drying (HAD) kinetics (50 ºC, 2 m/s) were carried out in a convective drier, which has already
been described in the literature [1]. Atmospheric freeze drying (AFD) experiments (-14±1 ºC, 2 m/s) were
conducted in a convective drier with air recirculation, which was placed inside a freezing chamber to keep
the sample at low temperature. Vacuum freeze drying (VFD) cubes were obtained using a vacuum freeze
drier (Telstar, Lioalfa-6, Germany) that worked at 10-3 mbar and -45 ºC. In all the cases, HAD, AFD and
VFD experiments were carried out, at least, in triplicate and extended until samples lost 90 % of the
initial weight. Cubic particles (10 mm side) were obtained from the flesh of Spanish origin eggplants
(Solanum melongena var. Black Enorma). The rehydration experiments (HAD, AFD and VFD) were
carried out, at least three replicates, in distilled water at 25 ºC, until achieving constant weight. Textural
properties of dried/rehydrated and fresh eggplant cubes were assessed by texture profile analysis (TPA).
At least, 10 measurements were performed for each set of samples (HAD, AFD, VFD rehydrated and
fresh eggplant cubes). ANOVA (p<0.05) was carried out in order to estimate the influence of the drying
method on the hardness of the rehydrated samples.
A diffusion model based on the Fick’s law, which neglected the external resistance to mass transfer, was
used to mathematically describe the drying kinetics (HAD and AFD) of eggplant cubes. Rehydration
kinetics of the HAD, AFD and VFD eggplant cubes were also modeled considering the diffusion theory.
The effective moisture diffusivity (and also the equilibrium moisture for the rehydration kinetics) was
identified by minimizing the sum of the squared difference between experimental and calculated average
moisture content of samples. Identification was carried out by using the Generalized Reduced Gradient
(GRG) method, available in Microsoft ExcelTM spreadsheet from MS Office 2007. The goodness of the fit
was determined by calculating the percentage of explained variance (%VAR).

11th International Congress on Engineering and Food - Athens, Greece, 2011 1997
RESULTS & DISCUSSION
The results of the drying kinetics modelling are shown in Table 1. The value of the effective moisture
diffusivity (De) for hot air drying experiments was 7.64·10-10 m2/s and the percentage of explained
variance of the model 89.30%. The low value of the explained variance could mean that assumptions
considered in the model formulation were not properly chosen. The value of De for the atmospheric freeze
drying experiments was almost one order of magnitude lower (5.28·10-11 m2/s) than in HAD (Table 1).
The %VAR was 92.07%, showing that the diffusion model fitted to the AFD experimental data slightly
better than to HAD data.
The rehydration kinetics of the HAD and AFD eggplant cubes showed that AFD samples rehydrated
faster than hot air dried cubes and reached higher values of equilibrium moisture (Weq). Vacuum freeze
dried cubes reached the equilibrium moisture immediately after their immersion in distilled water (in less
than 2 seconds). This fact may be explained due to the high porosity of vacuum freeze dried samples. The
equilibrium moisture reached by the AFD and the VFD samples was similar and close to the fresh
eggplant (12.285±0.925 kg w/kg d.m.).
Table 1 also shows the results of the rehydration kinetics modelling, that is the effective moisture
diffusivity, the equilibrium moisture content and the percentage of explained variance. The diffusion
model fitted to the HAD rehydration kinetics better than to the AFD ones. Moreover, the effective
moisture diffusivity for the AFD rehydration experiments was almost one order of magnitude higher than
in HAD experiments due to the high porosity of the AFD cubes. Hot air drying reduces the intercellular
spaces and creates a compact tissue, partially losing the spongy structure of the eggplant and decreasing
the porosity [2]. In AFD samples, the shrinkage was almost negligible due to drying being conducted at a
temperature lower than sample thawing point, thus a very low cellular stress is produced.
Table 1. Results of drying and rehydration kinetics modelling.
De (m2/s) Weq (kg w/kg d.m.) VAR (%)
Drying HAD 7.64·10-10 - 89.30
AFD 5.15·10-11 - 90.37
-9
Rehydration HAD 4.43·10 6.496 96.56
AFD 2.56·10-8 9.653 88.58
Dehydration involves a high degradation of eggplant structure [2], thus rehydrated samples don’t recover
the initial texture. Regardless the drying method, dried/rehydrated samples were much softer than fresh
eggplant. Significant differences (p<0.05) between the texture of dried samples were also observed. VFD
samples showed the lowest and HAD the highest hardness after rehydration. AFD samples showed
intermediate textural properties.

CONCLUSION
Drying method prompted changes in textural properties, as well as, rehydration pattern of eggplant
samples. AFD samples showed an intermediate rehydration rate and hardness between HAD and VFD
samples. Therefore, AFD could represent an interesting alternative to HAD and VFD to achieve high
quality dried products with lower cost than VFD.

REFERENCES
[1] García-Pérez J.V., Cárcel J.A., de la Fuente S. & Riera E. 2010. Ultrasonic drying of foodstuff in a fluidized bed:
parametric study. Ultrasonics, 44, e539-e543. [2] García-Pérez J.V., Puig A., Pérez-Munuera I., Cárcel J.A. & Riera E.
2010. Kinetic and microstructural changes induced by power ultrasound application on convective drying of eggplant.
Proceedings of 20th International Congress on Acoustics, ICA 2010, 23-27.

1998
 Effect of Fluidized-bed drying on the microstructure of higuerilla seeds (Ricinus

communis). An alternative source of protein and biofuel.

José Jorge Chanona Péreza*, Eduardo Terrés Rojasb, Jorge Alberto. Mendoza Pérezc, Hilda María
Hernándeza, Gustavo Fidel Gutiérrez Lópeza, Vicente Garibay Feblesb and Maria de Jesús Perea Floresa

a
Departamento de Graduados e Investigación en Alimentos, Escuela Nacional de Ciencias Biológicas,
Instituto Politécnico Nacional, México D.F.( jorge_chanona@hotmail.com )
b
Laboratorio de Microscopía Electrónica de Ultra Alta Resolución. Instituto Mexicano del Petróleo.
México D.F.
c
Departamento de Ingeniería en Sistemas Ambientales, Escuela Nacional de Ciencias Biológicas,
Instituto Politécnico Nacional, México D.F.

INTRODUCTION
Drying process induces microstructural changes that affect the physical properties and
functionality of biomaterials; for example, the oil extraction could be improved and the toxic
compound inactivated. Higuerrilla seed (Ricinus communis) is an interesting material due to
their large content of proteins and lipids [1]. Thus, the oilseed can be extracted and converted
to biodiesel and the proteins remain of oil extraction could be used for human or animal
consume, if the toxic substances are inactivated during thermal process. Fluidized bed drying
can be useful to induce changes on the structure and functionality of R. communis seed such as
coat cracking, damage in the endosperm cells and inactivation of toxins, these structural
changes in the seeds could be benefic for their use as source of the protein and biofuel [2]. The
aim of this work was to evaluate the microstructural changes during convective drying of R.
communis by means of microscopy techniques and image analysis.

MATERIALS & METHODS

R. communis seeds variety Silvestre Tiripiteo were kindly provided by means Promotora
Agricola “La Estancia” (Michoacán, México). Seeds without fissures and damage were
selected and cleaned manually. Next, R. communis seeds were dried in fluidized-bed at
different temperatures (80, 90, 100, 110 ºC) at constant air flow (7 m/s). Effective Diffusion
Coefficient of water vapor (Deff) and Activation Energy (Ea) were measure from drying
kinetics by means of Second Fick law and Arrehnius equation.
The effect of drying process on oilseed structure was evaluated by using of Environmental
Scanning Electron Microscopy (ESEM) and Image Analysis (IA). For ESEM observations,
thin slices (45 Pm) of endosperm tissue were obtained with a freezing microtome (Leica, 157
CM1850, Germany) and seed coat was observed directly in the electronic microscope. The
samples were mounted on aluminium stubs whit carbon adhesive tape and directly observed by
XL-30 ESEM (Philips, USA) at 25 kV accelerating voltage. For IA, the images obtained were
used to extracted cellular shrinkage from ESEM images of endosperm and cranking of seed
coat was observed.

11th International Congress on Engineering and Food - Athens, Greece, 2011 1999
RESULTS & DISCUSSION
The final moisture content of dried oilseed oscillated between of 1.43-0.6% wet basis in an
interval of 80-110 °C. The Deff values were affected by the increment air temperature (6.3x10-
10
–3.2x10-9 m2/s from 80 to 110°C), additionally an Ea of 55.9kJ/mol (R2=0.87) was obtained
by fitting of the Deff values with the Arrhenius equation. Drying kinetics (Figure 1) showed
that the water diffusion was controlled mainly by the internal structure of the material and the
water is strongly binding to the structure of the material. This fact, it has been reported for
similar drying seeds studies [3]. In this curves, there was not constant-rate period but it seen to
occur the falling-rate period. Endosperm

1.0

S/T
0.8

0.6
Xt/XO

80°C (7m/s) 80㼻C

90°C (7m/s)
0.4
100°C (7m/s)
110°C (7m/s)

0.2

110㼻C
0.0

0 50 100 150 200

tiempo (min)

Figure 1. Experimental drying kinetics of Figure 2. Gallery of ESEM micrographs of the

R. communis seeds at different temperatures effect of Fluidized-bed drying on the endosperm
of R. communis seeds at different temperatures.

The micrographs (Figure 2) showed structural damages on the endosperm at 80°C and IA
allowed evaluate a 13% of cellular shrinkage in endosperm cells, in contrast a complete
destruction of endosperm was obtained in the material drying at 110°C, and the cellular
shrinkage cannot be evaluated. The seed coat showed a major damage when the material was
drying at 110 ºC. The cellular damage induced by drying process in the endosperm of seeds
could help to oil diffusion and to improve the oil extraction yield. The oil obtained from seed
could have a potential use in the production of biodiesel. Structural changes observed could
explain the effect of the drying conditions on the material structure.

CONCLUSION
The drying kinetics were controlled by internal microstructure of the biomaterial. The cellular
damage and low moisture caused by drying process in the endosperm of seeds could help to oil
diffusion and with it improve the oil extraction yield.

REFERENCES
[1] Ogunniyi, D.S. 2006. Castor oil: A vital industrial raw material. Bioresource Technology. 97, 1086-
1091.
[2] Aguilera, J.M., Stanley, D.W. (1999). Microstructural Principles of Food Processing and Engineering.
2nd Ed. Food Engineering Series. Gustavo V. Barbosa-Cánovas, Series editor. Aspen Publishers, Inc.
[3] Sablani, S.S., Rahman, M.S. 2008. Fundamentals of Food Dehydratation in FOOD DRYING Science
and Technology: Microbiology, Chemistry, Applications. Edited by: Hui, Y.H., Clary, C., Farid,
M.M., Fasina, O.O., Noomhorm, A., Welti-Chanes. DEStech Publications, Inc. Pennsylvania, U.S.A.
1-42.

2000
A simple mathematical model proposed to predict kinetics of mass transfer in osmotic dehydration

of muskmelon

Jose Lucena Barbosa Jra,c, Daniel Guimaraes Correa Moreira Rochaa, Maria Ivone Martins Jacintho
Barbosaa, Mauricio Cordeiro Mancinib, Miriam Dupas Hubingerc
a
IT/UFRRJ, Department of Food Technology, Seropedica-RJ, Brazil (lucena@ufrrj.br)
b
IT/UFRRJ, Department of Chemical Engineering, Seropedica-RJ, Brazil (mancini@ufrrj.br)
c
FEA/UNICAMP, Department of Food Engineering, Campinas-SP, Brazil (hubinger@fea.unicamp.br)

INTRODUCTION
There is a lot of published information describing the influence of variables on mass transfer rates and
considerable effort has been made toward developing models to predict the mass transfer kinetics of
osmotic dehydration (OD). Due to the simplifications involved in solving the diffusion equation for a
food subjected to osmotic process, the Fickian diffusion model becomes an empirical equation, despite its
theoretical basis. So, it should be used very carefully. Therefore empirical models are important, notably
for non-classical geometries, whose solutions of the diffusion coefficients involve numerical methods.
Some authors have used statistical tools to study process variables, such as Response Surface
Methodology (RSM). However, information about the process kinetics is omitted in this type of approach
and moreover the process time could mask the effect of other process variables. Some efforts have been
made to overcome these limitations. Nevertheless, it is still difficult to define adequately the processing
time. The aim of this work was to propose a simple mathematical model to predict the water losses and
solute gain during osmotic process. The specific objective was the development of a correlation between
dehydration/impregnation rates with effective diffusivity. The applicability of the model was evaluated
for orange-fleshed muskmelons osmotically dehydrated in corn syrup solutions.

MATERIAL & METHODS

Samples and osmotic dehydration treatment
Fresh muskmelons were cut into slices (30 x 40 and 5 mm) and subjected to osmotic process for 30, 60,
90, 120, 180, 240, 360, 600 and 1440 minutes for each temperature (28 to 42 °C) and solute concentration
(38,7 to 61,3% w/w) condition, given by central composite experimental design, agitation (80 rpm) and
weight ratio of sample to solution (1:10) kept constant. Water loss (WL) (g water/100g initial wet sample)
and solids gain (SG) (g solids/100g initial wet sample) were determined by gravimetric method.
Kinetic models
Peleg modified model: The reciprocal value of Peleg rate constant (K1) is related to the drying rate at the
very beginning (i.e. t=0), suggesting a relationship between K1 and the driving force of the process. The
half-life for dehydration/impregnation rate (t1/2) can be expressed as follow:
dX (t ) 1 dX (t ) (1)
dt t t1 / 2 2 dt t 0

Thus, substituting into Peleg’s model, we obtain: (2)

t1
2
K1
K2
 
2 1

Data relative to mass transfer parameters were fitted using the equation proposed by Peleg modified in the
present work, in order to compare itself to the diffusion model.
Statistical Analysis: A first-order central composite rotatable design was used for designing the
experiments for osmotic dehydration of muskmelon slices using two factors. The obtained coefficients
were subjected to analysis of variance, test of lack of fit and the criterion used to evaluate the best fitting

11th International Congress on Engineering and Food - Athens, Greece, 2011 2001
model was their average relative error (P) less than or equal to 10% are considered to fit the experimental
data satisfactorily.

RESULTS & DISCUSSION

Both models showed low dispersion (P values under 10% and R2-values near 1.0), showing a good fitting
capability for WL and SG (Fig. 1). The observed difference between half-life for WL and SG was due to
the fact that the upcoming solids during osmosis take place mostly between the extracellular space and
not through the selective cell membrane as observed for water fluxes. It was obtained mass diffusivity
values ranging 1.11-2.25x10-10 for WL and 1.10-2.27x10-10 m2.s-1 for SG, respectively. It can be observed
that the proposed model is more accurate relative to diffusive model, mainly at the beginning of the
process. Furthermore, it has the advantage of allowing the calculation of the equilibrium values
satisfactorily. The lower fitting capacity of diffusive model relative to the other models has been reported
by several authors. This can be explained by that other phenomena besides diffusion are occurring during
the osmotic process, or the assumptions made to obtain diffusive model are not fulfilled, especially in
high viscosity solutions when high solids concentration was used.
1.00 The values of the half-life were higher for highest mass transfer
(a) potential, showing the relationship between dehydration and solid
predicted

0.75
uptake rates and mass transfer potential.
Analysis of Variables Influence on t1/2, dX(t)/dt|t=0 and X(t1/2)
0.50
for WL and SG
0.25 High correlation coefficients were obtained for WL’s models,
experimental
whereas for SG, a poor correlation for t1/2 was obtained and the
0.00 models for initial rate and solids gain at the half-life were not
0.00 0.25 0.50 0.75 1.00
1.00 predictive. Therefore, temperature and solution concentration
(b) showed no significant influence on solid uptake rates at the
predicted

0.75
beginning of the process. The variation in half-life values (higher
0.50
for SG) occurred around a mean value, providing a range of
processing time (30 to 70 minutes) in which the potential transfer
0.25 reduction is similar, regardless the type (dehydration or
experimental
impregnation) and conditions of process. In some researches,
0.00 osmotic dehydration process times were higher than 120 minutes
0.00 0.25 0.50 0.75 1.00
and an important observation to be considered in these cases is
Figure 1. Fitting capability of the
solids amounts incorporated to the fruit during the osmotic
diffusive (a) and proposed (b)
dehydration. If for quality reasons, the fruits amount of sugars has
models for WL(Ŷ) and SG(Ÿ)
to be enhanced, the period of time that the fruit should stay in the
osmotic solution needs to be higher. Alternatively, if the process focus is on minimal solids uptake and
cost reduction the period of time has to be as short as possible. It is important to remark that these models
have a limited validity within the experimental range which their coefficients were obtained.

CONCLUSION
The proposed model was able to predict the kinetics of osmotic dehydration and the WL and SG values at
equilibrium. A correlation between rates of dehydration/impregnation process and effective diffusivities
during osmotic dehydration of orange-fleshed muskmelon was obtained.

2002
 Drying characteristics of Açaí (Euterpe oleracea)

Antônio M. Barbosa Neto, Luanda G. Marques, Manoel M. Prado

Department of Chemical Engineering, Federal University of Sergipe, São Cristóvão-SE/Brazil

(manoelprado@ufs.br)

INTRODUCTION
Fresh açaí fruit is highly perishable and drying is a potential method to increase its shelf-life
for further use. However, to these authors knowledge, studies on drying of whole açaí berries
have not been reported.in the literature.
Due to their high initial moisture content açaí fruits tend to undergo volumetric changes upon
water removal, thus affecting heat and mass transfer. The changes in shape, dimension and
solid structure of deformable particles yield to a particular system where the available
coefficients of mass transfer are not suitable to reproduce moisture loss during drying of the
material. Thus any attempt to characterize the drying behavior of the açaí berries must address
the shrinkage characteristics. The shrinkage phenomenon affects the length of the diffusion
path in dried material, which influences the moisture diffusion coefficient of the material and,
as result, influences the drying rate [1].
In the present work, the effects of drying air temperature on drying kinetics and shrinkage of
whole açaí berries are presented and discussed, as well as values of moisture diffusivity
estimated with and without incorporating the shrinkage in diffusion model are compared and
analysed.

MATERIALS & METHODS

Açaí fruits from the “Black” cultivar, produced in Belém-PA, Brazil, were used in this
investigation. Convective drying of whole açaí fruits was carried out using a cross flow air
oven dryer at four different temperatures (40, 50, 60 and 70oC) and air velocity of 1.0 m/s. The
shrinkage of açaí berries during drying was quantified from the changes in volume and surface
area of individual berries, which were calculated from the dimensions taken by the digital
micrometer, assuming that açaí berries are oblate spheroids.
Shrinkage of individual particles was evaluated in terms of reduced dimensional change in both
volume (Vp/Vp0) and surface area (Ap/Ap0).
The mass transfer in drying of açaí fruits was analyzed in terms of effective diffusivity, which
was determined at each temperature by applying diffusional models of Fick’s second law for
sphere without and with consideration of shrinkage [2, 3] to describe the drying kinetics.

RESULTS & DISCUSSION

The surface area and volume of açaí berries decreased about 18 and 26% until the end of the
process, respectively, stressing the need to take into account the surface area changes to the
calculation of the water flux density as well as to include the shrinkage in the mass transfer
models.
Three distinct drying flux periods were identified during thin-layer drying of açaí berries: a
short heating-up period followed by two falling flux periods. No constant drying flux period
was observed, in spite of the exchange area correction. The pronounced period of falling flux

11th International Congress on Engineering and Food - Athens, Greece, 2011 2003
indicated that the moisture diffusion inside the solid was the main physical mechanism
governing mass transfer in drying.
Table 1 shows the results obtained for the effective diffusion coefficient estimated from
diffusional models without and with consideration of shrinkage, for the four drying air
temperatures investigated in this study. The obtained values are within the range of diffusivities
reported in the literature for fruits and vegetables [4]. It can be verified that the values of
diffusivity calculated without considering the shrinkage were higher than those obtained taking
into account the phenomenon. Neglecting shrinkage of individual particles during thin-layer
drying leads, therefore, to an overestimation of the mass transfer by diffusion. The explanation
is on the fact that berry shrinkage produces a variation in the distance required for the
movement of water molecules, therefore making that effective diffusivity be overestimated
when obtained from analytical solution analytical of diffusion without considering volume
contraction. This finding agrees with results of previous reports on other products [3].

Table 1. Effective coefficients of diffusion estimated without and with consideration of shrinkage during
açaí berries drying

CONCLUSION
Thin layer drying of açaí berries was characterized by significant area and volume changes
upon moisture removal. Changes in fruit volume and surface area were found to be dependent
only on the average moisture content of the whole fruit. The process was found to take place
predominantly in the falling flux period, indicating that the mass transfer was limited by
diffusion of moisture from inside to the surface of the material. Neglecting the shrinkage of
whole açaí fruits led to an overestimation of the effective diffusion coefficient and,
consequently, of the energy required for drying the material.

REFERENCES
[1]. Bialobrzewski, I.; Zielínska, M.; Mujumdar, A. S. & Markowski, M. 2008. Heat and mass transfer
during drying of a bed of shrinking particles – Simulation for carrot cubes dried in a spout-
fluidizedbed drier. Int. J. Heat Mass Transfer, 51, 4704–4716.
[2]. Crank, J. 1975. The mathematics of diffusion, 2nd, Ed., Clarendon Press, Oxford, London.
[3]. Souraki, B. A. & Mowla, D. 2008. Axial and radial moisture diffusivity in cylindrical fresh green
beans in a fluidized bed dryer with energy carrier: Modeling with and without shrinkage. Journal of
Food Engineering, 88, 9-19.
[4]. Ramos, I. N.; Miranda, J. M. R.; Brandão, T. R. S. & Silva, C. L. M. 2010. Estimation of water
diffusivity parameters on grape dynamic drying. Journal of Food Engineering, 97, 519-525.

2004
 Vitamin C Content of Freeze-Dried Tropical Fruits

Luanda G. Marquesa, Manoel M. Pradoa; José T. Freireb

a
Department of Chemical Engineering, Federal University of Sergipe, São Cristóvão, Brazil
(luanda_gimeno@yahoo.com.br)
b
Department of Chemical Engineering, Federal University of São Carlos, São Carlos, Brazil
(freire@power.ufscar.br)

INTRODUCTION
Brazil is among the largest producers of tropical fruits in the world. Tropical fruits such as
guava, mango, papaya and pineapple are excellent source of carotenoids and vitamin C. In
recent years, increasing attention has been paid to the role of diet in human health. Among
antioxidant vitamins, vitamin C has many biological activities in human body reducing the risk
of arteriosclerosis and some forms of cancer, a marker of inflammation and possibly a
predictor of heart disease [1, 2]. Commercialization of dried fruits has gained importance
worldly due to the search of consumers for practicability and products of great nutritional
value. The application of the freeze-drying technique in fruits conservation is the key for a
successful of their commercialization.

MATERIALS & METHODS

Fresh guava, mango, papaya and pineapple were purchased from local market in the city of São
Carlos-SP, Brazil. The fruits were cut into slices of 5 mm thickness and subsequently were
frozen in liquid N2. The frozen fruits were dehydrated in a laboratorial scale freeze-dryer,
manufactured by Edwards, L4KR model. The ascorbic acid content of “in nature”, freeze-dried
and hot air dried pulps was determined using the 2.6 dichlorophenolindophenol titration [3].

RESULTS & DISCUSSION

After freeze-drying the moisture content was equal to 5, 4, 2 and 7% (wet basis) for guava,
mango, papaya and pineapple, respectively. Figure 1 presents the vitamin C content determined
for “in nature” and freeze-dried tropical fruits. It can be observed after the process vitamin C
losses of 37.47, 3.05; 6.91, and 27.31% for guava, mango, papaya and pineapple, occurred,
respectively. The difference in ascorbic acid content of freeze-dried fruits can be explained by
the differences between the physical and chemical properties of morphological structures of
each fruit.
The results shown in Figure 1 are in agreement with those obtained by Marques et al. [4]. The
authors determined the vitamin C content in freeze-dried acerola for three different ripening
stages. It was observed that the smaller loss of vitamin C was of 13% for the intermediate stage
(yellow-reddish fruits), while the greater loss was of 69.3% for the green fruit. The vitamin C
losses for freeze-dried fruits are considerable smaller when compared the vitamin C losses
caused to others drying methods.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2005
 Figure 1. Vitamin C content for the fresh and freeze-dried tropical fruits.

Cutting and slicing fruits may induce a rapid enzymatic depletion due to the cellular disruption
which allows contacts of substrates and enzymes [1]. Thus, the vitamin C losses can be due not
only to primary and secondary drying of freeze-drying (thermal processing), but also by the
operations before drying such as cutting, slicing and freezing (first stage of freeze-drying).

CONCLUSION
The vitamin C of both fresh and freeze-dried guava, mango, papaya and pineapple was
determined. Although losses of 37.47; 3.05; 6.91, and 27.31% for guava, mango, papaya and
pineapple, respectively, have occurred, the freeze-dried tropical fruits were characterized by a
high nutritional value, when compared with those dried by other methods.

REFERENCES
[1] Podsedek, A. 2007. Natural antioxidants and antioxidant capacity of Brassica vegetables: A
review. LWT-Food Science and Technology, 40, 1-11.
[2] Lee, S. K. & Kader, A. A. 2000. Preharvest and postharvest factors influencing vitamin C
content of horticultural crops. Postharvest Biology and Technology, 20, 207-220.
[3] Benassi, M. T. & Antunes, A. J. 1988. A comparison of metaphosphoric and oxalic acids as
extractants solutions for the determination of vitamin C in selected vegetables. Arq. Biol.
Technol., 31(4), 507-513.
[4] Marques, L. G.; Ferreira, M. C. & Freire, J. T. 2007. Freeze-drying of acerola (Malpighia
glabra L.). Chemical Engineering and Processing, 46, 451–457.

2006
 Rehydration characteristics of freeze-dried avocado (Persea americana)

D. S. Souzaa; J. D. R. Pimentela; M. M. Pradob; L. G. Marquesb;N. Naraina

a
Post-Graduate Program in Food Science and Technology (NUCTA)/ Federal University of Sergipe, São
Cristóvão-SE, Brazil (narendra.narain@gmail.com)
b
Department of Chemical Engineering/Federal University of Sergipe, São Cristóvão-SE, Brazil
(luanda_gimeno@yahoo.com.br)

INTRODUCTION
Avocado (Persea americana Mill.)is a commercially valuable crop whose trees and fruits are
cultivated in tropical climates throughout the world. Approximately 75% of an avocado’s
calories come from fat, most of which is monounsaturated fat [1]. Several researchers have
attempted to obtain stable avocado pulp, using a series of preservation methods. Dried products
are usually rehydrated prior to their use. Rehydration is a complex process aimed at the
restoration of the properties of the raw product. The aim of this work was to evaluate the
rehydration capacity of freeze-dried avocado pulp powder and pieces. Peleg and Weibull
equations were used to describe the rehydration kinetics. The hardness of fresh, freeze-dried
and rehydrated avocado was measured using a texture analyser.

MATERIALS & METHODS

The avocado fruits (variety Collinson) were purchased in the city of Aracaju - SE. In order to
assess the influence of different geometries on the rehydration process, the samples were cut in
the shape of cubes, slabs and disks. Samples of beaten avocado pulp were also prepared. After
freezing, the samples were dried in a laboratorial scale freeze-dryer. Lyophilized avocado
powder was rehydrated in a equipment similar to that proposed by Martins & Pinto [2], while
freeze-dried avocado slices were rehydrated by immersing them in a water bath at room
temperature [3]. The equations proposed by Peleg and Weibull, which can be given by,
respectively:

where k1 is a kinetic constant with dimension of time and k2 is a dimensionless parameter,

which is related to the maximum moisture content at saturation, Xe; X0 is the initial moisture
content (d. b.), X is the moisture content at time (d. b.); and \is the shape parameter
(dimensionless), Tis the scale parameter, with dimension of time. Tests on texture were carried
out with fresh, freeze-dried and rehydrated samples at room temperature using a Texture
Analyser.

RESULTS & DISCUSSION

Figure 1 shows the rehydration kinetics of freeze-dried avocado pieces of different geometries
and the fit of the Weibull equation. It may be noted that the structure in which the samples
were lyophilized influenced the rehydration process. The curves showed a typical behaviour of

11th International Congress on Engineering and Food - Athens, Greece, 2011 2007
rehydration with a fast initial water absorption period followed by a slower rate in the later
stages. It can be seen that the equilibrium moisture content at saturation does not reach the
moisture content of the raw avocado, indicating that changes using the freeze-drying process
are irreversible. However, comparing these results with the avocado dried using tray-dryer [4],
it is evident that the freeze-drying produces a best product.

Figure 1. Moisture content in dry basis as a function of time during rehydration of freeze-dried avocado

According to Lee et al. [4], it is possible that the cube samples did not induced significant
changes in the internal alignment of avocado cell structure. On the other hand, the disk and slab
samples could have caused damages in this cell structure, which acted as internal resistance to
the water flow during rehydration.

CONCLUSION
The powder obtained from freeze-dried beaten pulp and avocado cube presented the higher
rehydration rates and moisture contents at saturation. The Weibull equation was found to be the
most suitable to describe the rehydration behaviour of freeze-dried avocado in all geometries
investigated in this work. The texture of avocado was affected by both the lyophilization and
rehydration processes as well as by the sample geometry.

REFERENCES
[1]. Fernandes, F. A. N.; Rodrigues, S.; Law, C. L. & Mujumdar, A. S. 2010. Drying of Exotic Tropical
Fruits: A Comprehensive Review. Food Bioprocess Technol, DOI 10.1007/s11947-010-0323-7,
published online, 24 February.
[2]. Martins, P. C. & Pinto, L. A. A. 2003. Caracterização da secagem de cebola (Allium cepa L) em
camada delgada e da reidratação do produto desidratado. Brazilian Journal of Food Technology,
6(3), 144-151.
[3]. Marques, L. G.; Prado, M. M. & Freire, J. T. 2009. Rehydration characteristics of freeze-dried
tropical fruits. LWT-Food Science and Technology, 42, 1232-1237.
[4]. Lee, K. T.; Farid, M. & Nguang, S. K. 2006. The mathematical modelling of the rehydration
characteristics of fruits. Journal of Food Engineering, 72, 16–23.

2008
 Studies on the cooking conditions and mechanical koji-making of black beans

Chia-Ling Jaoa, Wen-Ching Kob, Kuo-Chiang Hsuc

a
Department of Food Science and Technology, Tung-Fang Design University, Kaohsiung, Taiwan,
(e0306@ms7.hinet.net)
b
Department of Bioindustry Technology, DaYeh University, ChangHua, Taiwan,
(wcko@mail.dyu.edu.tw)
c
Department of Nutrition, China Medical University, Taichung, Taiwan, (kchsu@mail.cmu.edu.tw)

INTRODUCTION
Inyu is a traditionally fermented food product in Taiwan. It is made from black bean through
soaking with water, cooking, fungus inoculating, artificial koji-making, koji-washing,
incubating, salt adding and outdoor fermenting. Though Inyu has special properties in flavours
and qualities, its market share is lower than soy sauce made from soybean with wheat by
automatic koji-making due to limitation in production scale and labour cost. So, to simplify
and ameliorate the Inyu making procedure is necessary. In this study, the physical and
chemical properties of the cooked black beans designedly treated at different soaking time,
cooking temperature and time by using response surface methodology (RSM) was determined
to obtain the optimum soaking and cooking conditions. Furthermore, mechanically koji-
making method in different thickness of the cooked beans was investigated to evaluate the
feasibility of koji-making method for Inyu.

MATERIALS & METHODS

Black beans were soaked with 10-fold volume water at 25ɗ for 2, 4 or 6 h, then cooked at 116,
124 or 132ɗ, for 15, 23 or 31 min. The physical and chemical properties of the cooked black
beans were used as the parameters to obtain the optimum conditions by RSM. Furthermore,
mechanical koji making was performed with Aspergillus oryzae in different thickness of the
cooked black beans (6, 12 and 18 cm) at 32±2ɗ and 95%RH to determine the amino acid
profiles and proximate compositions.

RESULTS & DISCUSSION

The moisture content and hardness of the cooked black beans increased and decreased,
respectively, with the soaking time and cooking time elongated, however, they were affected
insignificantly (p烍0.05) by cooking temperature. The cooking process during Inyu production
is for protein denaturation and starch gelatinization of black beans, and the denatured proteins
and the gelatinized starches can be digested by the protease and Į-, ȕ-amylase from A. oryzae.
Therefore, the enzyme susceptibility of the cooked black bean is an important parameter for the
determination of optimum conditions of soaking and cooking process [1, 2]. The protease
susceptibility reached the highest value at the soaking time of 4 h, cooking temperature of
116ɗ and cooking time of 23 min [3]. The Į- and ȕ-amylase susceptibilities of the cooked
black beans were significantly (p烋0.05) affected by cooking temperature and reached the
highest value at 132ɗ. Therefore, the optimum conditions for soaking and cooking of black
beans were 4 h for soaking time, 23 min for cooking time and 132ɗ for cooking temperature.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2009
At this condition, the moisture content and hardness of the cooked black beans were 50.7% and
61.6 g, respectively.
The protease and amylase activities of the koji reached the highest value of 525.3 and 1127
unit/g dry koji, respectively with the cooked black bean thickness at 12 cm after the 48-hr koji
making, and those from the mechanical koji-making method were both higher than the
traditional method. The proximate compositions of the raw Inyu made from traditional and
mechanical koji making were insignificantly (p烍0.05) different, however, the glutamic acid
content was higher in the raw Inyu made from mechanical koji making than traditional making.

Cooking temperature (ɗ)

116ɗ 124ɗ 132ɗ

15------------------23----------------31

1.00 1.00 1.00

0.80 0.80 0.80

0.60 0.60 0.60

Cooking time (min)

0.40 0.40 0.40

0.20 0.20
0.20

0.00 0.00
0.00

-0.20
-0.20 -0.20

-0.40
-0.40 -0.40

-0.60
-0.60 -0.60

-0.80
-0.80 -0.80

-1.00
-1.00 -1.00 -0.80 -0.60 -0.40 -0.20 0.00 0.20 0.40 0.60 0.80 1.00
-1.00
-1.00 -0.80 -0.60 -0.40 -0.20 0.00 0.20 0.40 0.60 0.80 1.00
-1.00 -0.80 -0.60 -0.40 -0.20 0.00 0.20 0.40 0.60 0.80 1.00

2-------------------4-----------------6 2------------------4------------------6 2------------------4-----------------6

Soaking time (hr)
Figure 1 Contour plot of protease susceptibility (mg formalin nitrogen / g dry sample)
responded with soaking time and cooking time

CONCLUSION
The optimum conditions for making cooked black beans were at the soaking time of 4 h,
cooking temperature of 132ɗ and cooking time of 23 min. The mechanically koji-making
method should be performed as the 12-cm thickness of cooked black beans fermented for 48 h.
At this condition, the proximate compositions of the raw Inyu were similar to those made by
traditional method. The mechanical method could increase 3 or 4-fold quantity of koji making
and save labour cost as compared to the traditional method.

REFERENCES
[1] Fumiyosi, M., Hideyuki, Y., Hideki, N. & Enjiro, K. 1990. Influence of soybean-wheat ratio and
water content in koji on the quality of matured koji and its soy-mash. Journal of Japanese Soy Sauce
Research Institution, 16, 133-137.
[2] Yong, F.M. & Wood, B.J.B. 1977. Biochemical changes in experimental soy sauce koji. Journal of
Food Technology, 12, 163-175.
[3] Yokotsuka, T. 1986. Soy sauce biochemistry. Advanced Food Research, 30, 195-329.

2010
 The use of xylanase to improve physicochemical characteristics of nixtamalized corn

flour and tortilla texture obtained by extrusion

Benjamín. Ramírez-Wongb,f, Luis C. Platt-Luceroa,b, Patricia I. Torres-Chávezb, Jaime López-Cervantesa,

Dalia I. Sánchez-Machadoa, Elizabeth Carvajal-Millánc, Fernando Martínez-Bustosd, Armando Quintero-
Ramose, and Ignacio Morales Rosasb

a
Instituto Tecnológico de Sonora. Cd. Obregón, Sonora, México.
b
Depto. de Investigación y Posgrado en Alimentos. Universidad de Sonora. Hermosillo, Sonora, México.
c
Centro de Investigación en Alimentación y Desarrollo, A,C. Hermosillo, Sonora, México.
d
Cinvestav Querétaro. Querétaro, México.
e
Universidad Autónoma de Chihuahua, Chihuahua, Chihuahua, México.
f
Corresponding author. Phone: (662) 2-59-22-07. E-mail: bramirez@guaymas.uson.mx.

INTRODUCTION
The softening of most corn pericarp layers is essential to form masa with acceptable sheeting
characteristics [3]. The use of the enzyme xylanase is a novel way to improve the production of
corn tortillas from extruded nixtamalized corn flour. Xylanase hydrolyzes the ȕ-1,4-glycosidic
bonds of xylose on the backbone of arabinoxylans. The cleavage produces arabinoxylans of
lower molecular weight that affect the physicochemical and rheological characteristics of masa
[1]. The objective of this study was to evaluate the effect of xylanase on the water absorption
index and water absorption capacity of ENCF. Moreover, the viscoelastic characteristics of
masa, the texture of resultant tortillas and an assessment of tortilla quality via a sensory panel
were investigated.

MATERIALS & METHODS

Commercial white corn, commercial hydrated lime powder and Grindamyl Powerbake 7500,
an uninhibited component of xylanase preparation from Bacillus subtilis, with an activity of
163 000 U/g were used. Samples of ground corn were blended in a mixer with 0.3% (w/w)
lime. Xylanase, previously diluted in water, was immediately added to the mixture to reach a
final moisture content of 30%. Xylanase concentrations used were 0, 0.05, 0.075 or 0.1%
(w/w). Extrusion was performed in a single-screw laboratory extruder. The extrusion zone
temperatures were 60 °C, 70 °C, 80 °C and 90 °C. Extrudates were dried. Then, the extrudates
were ground in a mill until 61-63% of the accumulated material had passed through the 80
mesh to obtain nixtamalized corn flour (ENCF). Tortillas were prepared with ENCF samples
(1.5 kg) with distilled water in accordance with the water absorption capacity (WAC). The
water absorption index (WAI) and WAC were determined on each ENCF. Moisture content
and viscoelastic characteristics (G', G'' and Tan į) were determined in the corn masa. Firmness
and rollability were measured at 2, 24 and 48 h after tortillas were made. Firmness was
measured using the Kramer’s Cell attached to a texture analyzer. Tortillas with the best
treatment (firmness) were evaluated by sensory evaluation. A completely randomized design
was used. An analysis of variance was performed for all data gathered. The Tukey test (P <
0.05) was used to compare specific treatments.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2011
RESULTS & DISCUSSION
The ENCF treated with xylanase showed significantly (P < 0.05) higher WAI and WAC than
the ENCF control. This may be attributed to the effect of xylanase on the arabinoxylans in the
extruded corn flour. The WAI of extruded flour without xylanase was similar to that reported
by Platt et al. [2]. The magnitude of Tan į increased with increasing treatments of xylanase in
comparison to the control without xylanase. This could indicate that corn masas became more
viscous in the presence of xylanase because it made the masa softer and changed the
viscoelastic properties. The Tan į values of ENCF at 0.075 and 0.1% xylanase showed no
statistical differences (P > 0.05). Tortilla firmness increased with storage time (Fig. 1). The
addition of xylanase decreased the rupture force value in tortillas as xylanase concentration
increased. The effect was more significant at 0.075 and 0.1% (w/w) xylanase. Tortillas with
these two levels of xylanase showed no statistical differences (P > 0.05), and they were 15%
less firm (softer) than the tortillas made from ENCF without enzyme. Tortillas made with the
extruded flour containing xylanase were more flexible, and they maintained high rollability
scores during storage. Corn tortillas made with ENCF containing xylanase enzyme had
acceptable organoleptic characteristics.

ENCF
ENCF with xylanase (0.05 %)
ENCF with xylanase (0.075 %)
ENCF with xylanase (0.1 %)
80

60
Firmness (kPa)

40

20

0
2 24 48

Storage Time (h)

Figure 1. Firmness of tortillas produced from extruded nixtamalized corn flours (ENCF) during storage.

CONCLUSION
The tortillas made from extruded flour with xylanase had a softer texture than tortillas made
without the enzyme.

REFERENCES
[1] Courtin C.M. & Delcour J.A. 2001. Relative activity of endoxylanases towards water-extractable and
water-inextractable arabinoxylan. Journal of Cereal Science, 33, 301-312.
[2] Platt-Lucero L.C., Ramírez-Wong, B., Torres-Chávez, P.I., López-Cervantes, J., Sánchez-Machado,
D.I., Reyes-Moreno, C., Millán-Carrillo, J., & Morales-Rosas, I. 2010. Improving textural
characteristics of tortillas by adding gums during extrusion to obtain nixtamalized corn flour. Journal
of Texture Studies, 41, 736-755.
[3] Serna-Saldivar S.O., Almeida-Dominguez H.D., Gomez M.H., Bockholt A.J., & Rooney L.W. 1991.
Methods to evaluate ease of pericarp removal on lime-cooked corn kernels. Crop Science, 31,842.

2012
 The design of non-contact automatic shell cutting machine of chestnut and the

investigation of its effect by means of chestnut shelling experiment

Hong-Wei Xiaoa, Zhi-Long Dub, Zheng Loua,Li-Hong Wanga, Jun-Wen Baia, Zhen-Jiang Gaoa *

a
College of Engineering, China Agricultural University, Beijing, China (hwxiao82@gmail.com)
b
Chinese Academy of Agricultural Mechanization Sciences, Beijing 100083 ,China (duzhilong88@163.com)
*Corresponding Author: Zhen-Jiang Gao, Professor, College of Engineering, China Agricultural University,
Beijing, China (zjgao@cau.edu.cn)

INTRODUCTION
Presently in China shell cutting of chestnut is done manually, which is tedious, time consuming and labor
intensive [1].To overcome the problems that occurred during chestnut shell cutting, the laser cutting
technique was applied to chestnut shell-cutting and a new chestnut shell cutting machine in the present
investigation. Another objective was to investigate the effect of the power intensity of laser and the
velocity of conveyor on the performance of chestnut shell cutting and to optimize the process parameters.
The Design Theoretical Basis
Laser cutting technique was used to replace the rotary blades during the chestnut shell cutting process. It
is known that laser is the high energy particles which were excited out of the high intensity of radiation.
This can make the power density and the temperature of the focus reach up to 107-1012W/cm2 and 104
ȭ, respectively [2]. Using the above characteristics of laser a new chestnut shell cutting machine was
designed, which formed traces in the chestnut shell that is the laser cutting scratches.
The Whole Structure and Working Principle
The machine was consisted by the vibration feeder, V-shaped belt, motor drive system, laser generator,
mirrors for laser reflection and focusing, control panel et al. The operating procedures are as follows: put
the chestnuts into the hopper; slide along the sloping trough to the V-shaped conveyor belt; adjusting the
velocity of conveyor and mirrors for laser reflection and focusing to cut open the chestnut shells and can’t
hurt the flesh.

MATERIALS & METHODS

Fresh chestnuts used in the present study were purchased from a local supermarket in Beijing, China,
which were harvested from Huairou a famous chestnut production area.
The experiments were carried out according to Table 1. The effects of two independent parameters power
intensity of laser (A) and velocity of conveyor (B) on the shelling rate as presented in Table 1 were
investigated using orthogonal design and range analysis [3]. The experiments were repeated twice and the
average values were used. The shelling rate was used to evaluate the cutting performance following the
methodology described by Gao et al. [1] with sligh modification. The cut chestnuts were spread in a
single layer in an impingement oven with 180oC and 8m/s as its shelling temperature and hot air velocity,
respectively. After 5 minutes the SR (shelling rate) was calculated using the following equation (1):
SR= shelling u 100% (1)
total

11th International Congress on Engineering and Food - Athens, Greece, 2011 2013
 Table 1. Experiments were scheduled and analyzed by orthogonal design method
Experiments Factor A Factor B Shelling rate (%)
number Laser power (W) velocity of conveyor (m/s)
1 40 0.06 55
2 60 0.06 96
3 80 0.06 98
4 40 0.12 20
5 60 0.12 67
6 80 0.12 97
7 40 0.18 5
8 60 0.18 24
9 80 0.18 73
K1 80 249 Summation of each
K2 187 184 factor index
K3 268 102
k1=K1/3 26.7 83 The average of
k2=K2/3 62.3 61.3 summation of each
k3=K3/3 89.3 34 factor index
Range analyse 62.6 49
Optimum A3 (80W) B1 (0.06m/s)
experimental
condition
Where KX=summation of each factor index, kx=the average of summation of each factor index=Kx/3

RESULTS AND DISCUSSION

From Table 1, it can be found that the shelling rate of cut chestnuts increased with the increase of the
laser power. When the velocity of chestnut conveyor was kept at 0.06m/s, the chestnut shelling rate was
55%, 96% and 98% at laser power of 40, 60, and 80W, respectively. The effect of the power intensity of
laser on chestnut shelling rate was more distinct than velocity of conveyor. It also can be observed that
the shelling rate decreased with increasing the transport velocity of chestnut. The optimum process
parameter was 80W and 0.12m/s, the chestnut shelling was about 97%.

CONCLUSION
The laser cutting technique was applied to chestnut shell-cutting and a new chestnut shell-cutting machine
has been designed. The experiments of chestnut shell-cutting using laser indicated that the chestnut
shelling rate increased with increasing of the laser power or decreased with increasing of transport
velocity.

REFERENCES
[1] Gao Z.J., Lin H., Xiao H.W. 2008 Air-impingement De-shelling of Chestnuts(C.mollisima):Process Parameter
Optimization. International Journal of Food Engineering, 4(2), Article 14.
[2] Yan J.X. 2004. Laser Priciples and Technology[M]. Beijing: Higher Education Press. (in Chiese)
[3] Yuan Z.F. & Zhou J.Y. 2000. Experimental design and analysis. Beijing: Higher Education Press (in Chinese).

2014
 Relationship between chromatographic profiling by HS-SPME and sensory quality of

mandarin juices: effect of squeeze technology

Rafael Alvarez Quinteroa, Catarina Passaro Carvalhob, Oscar Lara Guzmána, Julian Londono Londoñoa

a
Grupo de Investigación en Sustancias Bioactivas (GISB), Universidad de Antioquia, Medellín, Colombia
(malvarez153@gmail.com)
b
Corporación Colombiana de Investigación Agropecuaria (CORPOICA), C. I. La Selva, Rio Negro- Antioquia,
Colombia (cpassaro@gmail.com)

INTRODUCTION
The sensory quality is of great importance to the consumer and several studies have shown that the citrus
industrial process can affect chemical and sensorial properties of the juice industry. For citrus flavors,
there have been proposed impact compounds for some sensorial notes. For mandarin flavors, methyl-
Nanthranilate and thymol are associated with sensorial notes, with additional contributions from ȕ-pinene
and Ȗ-terpinene. In this study, we determine the effect of different squeeze technology commonly used in
tropical countries, on chemical and sensorial quality parameters of mandarin Clementine juice, and the
relationship between HS-SPME and sensory analysis.

MATERIALS & METHODS

Citrus fruits, essential oil and juice extraction
Fruits (Citrus clementina, Hort. Ex. Tanaka) with commercial maturity index were harvested at Támesis,
Colombia (5º 42'N, 75º 40'W, and 775m above see level) in March 2010. The fruits were randomly
divided into three lots of 100 fruit. Hydrodistillation of mandarin essential oil was performed by the
method from United States Pharmacopoeia (USP 25). The fruits were squeezed using two technologies:
(A) like Zumex® and (B) like FMC®. After squeezed, juices were refrigerated until analysis.
HS-SPME and GC/MS analysis
The juices were subjected to HS-SPME analysis using the fiber
divinylbenzene/carboxen/polydimethylsiloxane. Extraction time and temperature was 3 h and 40 ºC,
respectively. SPME fibber was inserted into the GC injector at 260 °C for 1 min. The volatile compounds
were analysed by a Agilent 7890 GC/MS 5975C, and two capillary columns: HP-1MS and HP-5MS.
Temperature of the oven was 200 ºC for 10 min, the injector temperature was 260 ºC and the carrier gas
was He (1.3 mL/min). The MSD temperatures of the ionization chamber and MS Quad were 230 and
150ºC, respectively. Mass spectra were obtained by automatic scanning at 4.51 scansí1 with energy
ionization 70 eV, in the mass range m/z 40–350. Identification of the components was based on the
comparison of their retention index (RI) and by matching with commercial mass spectral libraries
(NIST/EPA/NIH, 2008), The software AMDIS 2.68 and database
(http://www.pherobase.com/database/kovats/kovats-index.php).
Sensory evaluation
Sensory evaluation, was carried out by a trained sensory panel of eight women in a certified laboratory
(NTC 3884, same as ISO 8589). A sensorial profile by multidimensional approximation was established
based on the NTC 3925, 3501 and 3932. All descriptors were score on a 6-point category scale (0 = none
and 5 = strong); and general quality was score on a 3-point category scale (1 = low and 3 = high). For
each treatment three samples were evaluated, and three replicates for each treatment.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2015
RESULTS & DISCUSSION
All volatile compounds find for both squeezing technologies, have been previously reported for citrus,
nevertheless, striking differences were observed in volatiles recovered from juice squeezed with
technology A and B (Figure 1). For both technologies (A and B), the principal volatile components were
monoterpenes: cyclics (97.41–88.76%) and non cyclics (1.19-6.95%), no terpenic compounds (0.61–
2.71%) and sesquiterpenes (0.29–0.59%). The limonene content in the volatile fraction was 97.51% and
65.49% for juice (A) and (B), respectively. Other substances such as aldehydes, alcohols and
sesquiterpenes reported quantitative differences, and additionally some compounds were not detected. For
both juices, it is important to notice that methyl-N-anthranilate was not present and thymol was <0.001%,
key flavour-active compounds in mandarin juices. Additionally, the methyl-N-anthranilate was not
present either in the essential oil. It is not possible to say whether this was a result of differences related
with grow conditions in the tropic, time of season, metabolic alterations, or some other factors. Figure 2
show global scores for each descriptors. The juice (A), had high score for undesirable flavor descriptors
like acid (3.4/3.1), bitter (2.7/1.6), citrus peel (4.1/3.0), green (2.4/1.6) and spicy (2.6/1.5). The judges
described a feeling irritating when they tasted the juice (A), which was related to higher essential oil
content in this juice (head-space ratio between (A) and (B) was approx 9:1). This might explain the sharp
and negative response of the judges. Both results, chemical and sensorial analysis, agree that the
technology applied for produce juice can influence additional quality-related volatiles in citrus, like
previously reported for different processes in citrus industry.

Figure 1. Total ion chromatograms from A. mandarin Clementine essential oil; B. and C. juice squeezed with
technology (A) and (B) respectively, on HP5 column. One of the three replicate runs, which were virtually identical, is
presented.

CONCLUSION
Both squeezing technology, had low levels of active compounds like Thymol, and absence of methyl-
Nmethylanthranilate. Nevertheless, the quantitative proportion of components can assure a good sensory
score for juices, despite of essential oil content in the juice. There are important chemical and sensory
quality differences between two technologies, and there is a relationship between chemical and sensorial
analysis. This study suggests that the type of industrial process for produce juice affects the final flavour
quality of the product, what is crucial to decide which technology applies.

2016
Effect of magnetic fields and ultrasound on aerobic mesophiles and histamine in beef loin

tuna loin tuna (Thunnus albacares)

Víctor. Manuel. Gélvez, Ordóñez a, Lorenzo Fuentes Berrioa.

a
University de Pamplona-ab University of Cartagena-Colombia
E-mail: vmgelvez@unipamplona.edu.co – lfuentesb@unicartagena.edu.co
a
Group Research in Engineering and Food Technology: GINTAL - Faculty of Engineering
University of Pamplona- Colombia

INTRODUCTION
Tuna as well as other species of fish is a food of high nutritional quality, their proteins contain
all essential amino acids required by the Agency, it has an important content in vitamins and
minerals. Its consumption has been associated, with a decrease in cardiovascular accidents
attributed to the presence of polyunsaturated, fatty acids like Eicosapentaenoic Acid and
decosahexaenoic acid (Omega 3) [1].
It is well known that the tuna produces a series of compounds as histamine is prejudicial to
health, which is formed in the State post-mortem of the fish by the bacterial decarboxylation of
the amino acid Histidine. The affected fish are often those with a high content of belonging to
the family Scombridae (tuna and the macarellus) Histidine histamine content is an obstacle to
export species tunidas from the tropics and subtropics in international markets [2].

MATERIALS & METHODS

Procurement of raw materials
Raw filet tuna species Thunnus albacares, was supplied by the company Atunec S.A.,
Industrial area Barranquilla.
Application of high-intensity ultrasound.
Treatment by ultrasound (US) was used in a computer (Elmasonic E, 37 kHz), with regulation
of intensity and temperature, time as transmission medium used de-ionized water of high
purity, the trials were done in triplicate. Times of treatments were 0, 3, 4 and 5 minutes.
Application of magnetic fields
In treatment by static magnetic fields (MF) a computer (Scoli, 8A N240, R = 2.8 Ƿ ohm) was
used with an intensity of 1 Tesla, the trials were done in triplicate. Treatment times were 0.3, 4-
5 minutes.

RESULTS & DISCUSSION

Effect of treatment with ultrasound on aerobic mesophiles
Figure 1. Presents the effect of treatments with US on aerobic mesophiles counts in beef Loin
of tuna. Where control shows more content aerobic, Mesophiles that samples treated with US
where stands the effect of treatment on the microbiota. The statistical analysis conducted,
showed significant differences (p< 0,05) among the all the tuna meat samples. The above
results are attributed to the effect that generates the intracellular cavitation phenomenon which
causes inactivation or microbial destruction in feed, due to the damage in the cell walls of
micro-organisms [3],[4], and in the cytoplasmic membrane[5] [6] also reported shocks

11th International Congress on Engineering and Food - Athens, Greece, 2011 2017
generated by cavitation injured structural components and functional cell lysis of the same
extent.

Figure 1. Effect of ultrasound (37 KHz) on aerobic mesophiles in beef loin tuna

Effect of treatment with ultrasound on production of histamine

The determination of histamine production was effected in beef loin tuna loin tuna stored at 4 °
C for five 5 days, and the effect of the US is shown in table 1. Results show reduced histamine
production is proportional to the time of treatment and that there is significant difference
between each of the analyzed sample, which can be attributed to the phenomenon of cavitation,
which for very short time can reach temperatures of 5,500 ° C and pressures of 50 MPa [7].
And they agreed, [8], who claim the histamine in tuna flesh denatures treatments of 116 ° C for
90 minutes. The content of histamine in meat tuna after treatment with US for ever applied is
below the minimum values reported by the Social Security Ministry, which is100 mg/kg of
beef and fish [9].

Table 1. Shows the effect of ultrasound (37 KHz) in the production of histamine in beef loin tuna

CONCLUSIONS
Application of ultrasound, and magnetic fields for 3,4 or 5 min in tuna meat reduces
significantly (p < 0,05) its aerobic mesophiles flora and the production of histamine in the five
days of storage in cooling. Therefore, this kind of technology can be used as a means of
conservation of the fishery products.

REFERENCES
[1] FDA (Food and Drug Administration). (1998). Scombrotoxin (histamine) formation. In: Fish and fishery products
hazards and control guide (2nd ed., pp. 73–90). Washington, DC: Department of Health and Human Services, Public
Health Service, Nutrition, Office of Seafood. [2] Yang, T.S. (1999). A new high-intensity ultrasonic technology for
food dehydration. Drying Technology, 17, 597- 608. [3] Pothakamury, U., Barletta, B., Barbosa, G., Swanson, B.
(1993). Inactivación de microorganismos en alimentos usando campos magnéticos oscilantes. Revista Española de
Ciencia y Tecnología de Alimentos 33: 479-489.

2018
 Relationship between pectic substances and strand separation of cooked spaghetti

squash
a b a c c a
Kayoko Ishii , Ai Teramoto , Hiroko Kuwada , Yuri Jibu , Mayumi Tabuchi , Yasumi Kimura , Michiko
a
Fuchigami
a
Department of Nutrition and Life Science, Fukuyama University, Fukuyama, Japan
(ishii@fubac.fukuyama-u.ac.jp; kuwada@fubac.fukuyama-u.ac.jp; kimura@fubac.fukuyama-u.ac.jp;
fuchigam@fubac.fukuyama-u.ac.jp)
b
Department of Health and Nutrition, Kanto Gakuin University, Yokohama, Japan (teramoto@kanto-
gakuin.ac.jp)
c
Department of Nutritional Science, Okayama Prefectural University, Soja, Japan (yjibu@fhw.oka-
pu.ac.jp, tabuchi@fhw.oka-pu.ac.jp)

INTRODUCTION
Spaghetti squash (Cucurbita pepo L.) is one of the hard-shelled squashes in the cucurbit family
and typically grown as a winter squash. It is an American native vegetable. It is rugby ball
sized and oval-shaped and the rind is hard and ivory colored at maturity. Its center contains
many large squash seeds. It has a mild taste and crisp texture and may be boiled, steamed,
baked or microwaved. When cooked, the flesh can be pulled apart to form strands that
resemble spaghetti, hence its name.
Pectin is the main component of the middle lamella. It contributes to adhesion between
parenchyma cells of vegetables and mechanical strength of tissues. Maceration of vegetable
tissues seems to be brought about mainly by the degradation of pectin [1]. The softening of
vegetables during cooking is affected by the properties of pectic substances, especially the
degree of esterification [1][2]. The high methoxyl pectin easily broke down in hot neutral
solution and alkaline solutions by ȕ-elimination [3][4].
Therefore, the cause of the separation into strands during boiling of spaghetti squash seems to
be pectic substances which play a substantial role in the maintenance of intercellular cohesion,
especially high methoxyl pectin which breaks down by cooking. Thus, the purpose of this
paper is to investigate the relationship of spaghetti squash pectic substances and the separation
into strands during cooking.

MATERIALS & METHODS

Sample preparation, texture and structure measurements
Spaghetti squash was cut into 2 cm long pieces, peeled and the seeds discarded. Flesh samples
were dropped into boiling distilled water and cooked for 15 min, 30 min or soaked in 0.01N
HCl solution (pH 2.0) for 24 hrs at 35°C. Texture and histological structures of samples were
measured using a rheometer (NRM-2002J, Fudo Ltd.) and a cryo-scanning electron microscope
(S-4500, Hitachi Ltd.) , respectively.
Extraction of pectin and analysis of extracts
Pectic substances of raw and boiled samples were successively extracted as follows: 0.01N
HCl (at pH 2.0 and 35°C), 0.1M sodium acetate buffer (at pH 4.0 and 35°C) and 2% sodium
hexametaphosphate solution (at pH 4.0 and 90°C) [1][2]. These extracts were designated as

11th International Congress on Engineering and Food - Athens, Greece, 2011 2019
PA, PB and PC, respectively. The DEAE-cellulose column chromatography of PA, PB and PC,
extracted from both raw and 15 min cooked flesh, was performed by the same method reported
previously [1]. The degree of esterification of PA, PB and PC were determined using a gas-
chromatographic procedure [5].

RESULTS & DISCUSSION

Changes in texture and histological structure of spaghetti squash during cooking
Spaghetti squash flesh separated into strands when boiled or soaked in a 0.01N HCl solution of
pH 2.0. The shape of cells, which constituted strands, was different from the cells which
surrounded strands. The former was round and the latter was elongated. When cooked, the
shape of the former was maintained, but the latter, which contributed to adhesion between
strands, broke down. Thus, flesh separated into strands. After cooking for 15 min or soaking in
0.01N HCl solution, cell separation in the middle lamella was observed.
Changes in pectic substances of spaghetti squash during cooking
The amount of PA, PB and PC in raw samples was 237.4 mg, 99.6 mg and 7.3 mg / 100 g,
respectively. The degree of esterification (DE) of PA, PB and PC was 67.4%, 61.5% and
55.6%, respectively. The percentage of PA, which was high methoxyl pectin, was greatest.
Therefore, when squash was cooked for 15 min, about 50% of pectic substances were released,
perhaps because high methoxyl pectin was broken down by ȕ-elimination. The DE of spaghetti
squash was higher than DE of the other vegetables such as lotus, burdock and bamboo shoot.
Thus, the squash was more easily softened than lotus, burdock and bamboo shoot after
cooking. The amounts of cellulose, hemicellulose, lignin and pectin of spaghetti squash were
similar to Japanese radish root. This suggests that cellulose, hemicellulose and pectin
remaining in strands after cooking maintained crisp tender strands.

CONCLUSION
Spaghetti squash separated into strands when boiled or soaked in HCl solution of pH 2.0. High
methoxyl pectin was degraded by ȕ-elimination during boiling and extracted by soaking;
consequently, the flesh separated into strands. This suggests that high methoxyl pectin glues
cells together in the flesh of spaghetti squash.

REFERENCES
[1]. Fuchigami, M. 1987. Relationship Between Pectic Compositions and the Softening of the Texture of
Japanese Radish Roots During Cooking. Journal of Food Science, 52(5), 1317-1320.
[2]. Fuchigami, M. & Okamoto, K. 1984. Fractionation of Pectic Substances in Several Vegetables by
Successive Extraction with Dilute Hydrochloric Acid and Acetic Buffer Solutions. Journal of
Japanese Society for Nutrition and Food Science, 37(1), 57-64.
[3]. Albersheim, P., Neukom, H., & Duel, H. 1960. Splitting of Pectin Molecules in Neutral Solution.
Archives Biochemistry and Biophysics, 90, 46-51.
[4]. Neukom, H. & Deuel, H. 1960. Uber den Abbau von Pektinstoffen bei Alkalisher Reaction. Beiheft.
zu den Zeitschr. des Schweiz. Forstv. 30, 223-235.
[5]. Baltolome, L.B. & Hoff, J.E. 1972. Gas Chromatographic Methods for the Assay of Pectin
Methylesterase, Free Methanol, and Methoxy Group in Plant Tissues. Journal of Agricultural Food
Chemistry, 20(2), 262-266.

2020
 Improvement of an enzymatic process to elaborate orange segments in syrup

Robles-López R.a*, Dorantes-Nieto A.a, Díaz-Carvajal D.a, Robles-De la Torre R.R.a. Bibbins-Martínez
M.D.a.

a
Centro de Investigación en Biotecnología Aplicada CIBA–IPN, Tlaxcala, km 1.5 Carretera Tepetitla-
Tecuexcomac, Tepetitla de Lardizabal, CP 90700, México, *(mreynarobles@yahoo.com).

INTRODUCTION
The production of oranges potential in Mexico is in some places within the top worldwide
producers, but due to the lack of new technologies, it is necessary to generate higher value
added products. An alternative to orange processing is to obtain segments in syrup, which
significantly would increases its market value. There exists a variety of methods, to obtain the
orange segments in syrup, being the enzymatic method, the most technically and economic
viable. Hence the importance of this project, whose main objective is to improve the enzymatic
process for orange segments elaboration, giving sensorial properties resembles those of fresh
fruit.

MATERIALS & METHODS

The orange fruit was characterized physically and chemically, being the maturity degree 4, the
proposed to obtain complete segments with an ideal degree of acidity and sweetness. To
remove the flavedo, thermal (5, 3, y 1.5 min 90-93ºC), mechanic (manually with the help of a
knife), chemical (HCl 1%, NaOH 2%, 93ºC, 1 min, 2 min and 5 min) and enzymatic methods
were evaluated.
Albedo elimination. The oranges without flavedo were segmented manually and then chemical,
mechanic and enzymatic methods were evaluated with this purpose. The chemical one,
included incubation in HCl diluted for 20 min at room temperature, 0.75 % NaOH, 20 min, at
55-60ºC and 0.5% sodium bicarbonate. The enzymatic method at pH constant of 5.0 and
different enzymes (PeelzymTM I, IV, Rapidase PrimaTM , and Macerex(R), concentrations (0.1,
0.5, 1, 2, 5 and 10 % v/v), and times (30 to 60 min) were used. sugar concentration was
measured at the beginning and at the end of albedo elimination. Thermal inactivation was used
in enzymatic treatment, the residual activity was evaluated spectrophotometrically at 235nm.

RESULTS & DISCUSSION

For flabedo elimination, thermal with pure water at 93°C for 1.5 min was the best treatment
maintaining the fruit without damage and keeping textural and flavor characteristics, (Table 1),
the control of temperature is the most useful and important factor for minimizing the defects of
quality. On the other side, chemical 1 minute treatment at the same conditions was not enough
to facilitate orange peeling and 5 minutes treatment destroyed the fruit. For albedo elimination
the 0.5% enzymatic treatment was the best maintaining the sugars concentration, sensorial
properties contrasting with chemical method which showed more time consuming, and
affecting the flavor.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2021
 Table 1. Resume of results of the different methods for flavedo elimination
Treatment Albedo elimination Elimination of chemist residues
H O 90°C; 1.5min Easy There are not chemic contaminant
2
5% NaOH 100ºC, 1 min Hard With diluted acid
5% NaOH 100ºC, 2 min Very easy With diluted acid, and cold water
5% NaOH 100ºC, 5 min Broken up With diluted acid and cold water
3% Enzyme pH 3.0, 120 min Easy Thermal treatment and cold water, expensive

According with the sensorial analysis, syrup with 20°Bx, pH 3.5 and 0.05 % sodium benzoate
was the best in order to conserve the sensorial characteristics without fresh fruit flavor loss in a
period of three mounts.
The orange segments, in boiling point and light syrup stored at 4ºC in glass jars, were more
acceptable with respect to the sterilization method and the commercial one. The strong syrup
treatment was pleasant but much sweeter than natural fruit. No microbial activity was present
in the products storage at 4ºC after 120 days. Figure 1 shows, segments of the enzimatically
treated segments and the appearance of the bottled product.

Figure 1. Enzymatically treated segments and Orange segments in syrup

CONCLUSION
For flavedo elimination, thermal (95°C) treatment was the best. For albedo elimination 0.5 %
enzymatic complex at 40ºC by 40 minutes treatment gave best results and kept flavor and sugar
concentrations. Sensorial analysis has shown that developed product received more
acceptability and 100% preference compared to the commercial one.

REFERENCES
[1] Janser. E. 1995. Enzymatic Peeling of Fruit. En: Fruit Processing Symposium. Parma.
[2] Romajaro. F., Riquelme F., Pretel, M.A., Martínez, G., Serrano, M., Martínez, C., Lozano, P. Segura
P. y Luna, P.A. 1996. Nuevas tecnologías de conservación de frutas y hortalizas. Ediciones Mundi-
Prensa. España. 5: 133- 191.
[3] Torricella. R. G., E. Zamora y H. Pulido. Evaluación sensorial aplicada a la investigación, desarrollo y
control de la calidad de la industria alimentaria. Instituto de investigación para la industria
alimentaria. La Habana. Cuba. 7: 57- 97.

2022
 The Technology of Butters’ enriching with carrots’ Powder

Ɍ.Ɉ Rashevskaa professor, Ɉ.Ɇ. Vashekab , assistant

a - National University of Food technologies, Kiev, Ukraine, E-mail: rashevsk@nuft.edu.ua
b - National University of Food technologies, Kiev, Ukraine E-mail: Oksana.Vasheka@meta.ua

INTRODUCTION
The main scientists of the world consider that the situation of bad health and appearing some chronic
deseases in many cases is associated with bad nutrition. Because of this situation one of the main
problems nowadays is new products producing that are reach for biological active substances. It should be
a harmoniums combination of traditional food with natural additions. On the basic of conventional dairy
lines we developed the technologies of functional types of butter with herbal additions. We also provide
insertion additives to the finished butter during its mechanical treatment. The method consist of insertion
the specially prepared suspension of the herbal into the butter during its mechanical treatment. The
selection criteria of powders were the functional characteristics and harmonious combination with the
taste of the product. Immunemodulatory, oncology and radioprotective properties of carrot are well-
known and widely used around the world. That is why, the new type of butter with carrot powder was
developed.

MATERIALS & METHODS

The subject of research for the microstructure’s study were water suspensions of carrot powders, obtained
by thermal, convection and cold spray drying (powder Karotte-100 made by "OBIPEKTIN AG, CH-9220
BISCHOFSZELL", Switzerland). For making carrot’s microscopic suspensions’ preparation the powder
was restored in water at 30 ... 35 ° C. From the received suspensions a preparation was made for viewing
on an optical microscope MIN-8 with light “on passing" by the standard procedure. Were investigated the
samples of butter with carrot powder of convective and cold spray drying. The test samples were made by
the described technology of enriched butter producing. The powder influence on the phase transformation
in butter’s fat has been studied by differential scanning calorimetry methods.

RESULTS & DISCUSSION

We analyzed the microstructure of water solution of the additives. It was established that suspension of
the powder, produced by cold spray drying contains big and small parts (15-80 micrometers, 1-5
micrometers). Microstructure of recovered particles are similar to particles of fresh vegetable. The
suspension of powder, produced by thermal drying contains remains of destroyed tissues. Puring swelling
they stick together. The suspension of powder, produced by convectional drying contains recovered parts
with size – 15 micrometers.
The study of phase transformations in milk fat of butter samples was performed by using the thermal
differential scanning calorimeter (DSC). DSC curves showed the melting peaks of group’s of high-
melting (HMG), medium-melting glycerides (MMG) and a peak of compatible liquid phase and low-melt
glycerides (LMG). The curves showed their melting temperature (Figure 1). In Figure 1 the DSC curves
of just prepared samples of butter. In curve (Figure 1 I, a) the control sample we clearly see the interval
with a peak melting - 32,4 ° C, which corresponds to the group HMG. Melting peak at 13,2 ° C is in the
temperature interval of 8 ° C to 25 ° C and corresponds to a compatible melting compatible of MMG and
LMG. The largest peak temperature of 3,8 ° C was formed by melting of LMG and liquid phase of
product. In the curve the jump of devitrifycation at -31 ° C is marked. The process of devitrifycation

11th International Congress on Engineering and Food - Athens, Greece, 2011 2023
passes without change of phase, and therefore it is not a phase transition. During this process the process
of "unfreezing" of mobility glycerides of milk fat is going on.
When comparing the DSC curves of fresh made butter
with the carrot powder of convective drying and the
control sample, we can see that the introduction of
additives leads to the appearance of distinct melting
peaks diffuse of MMG and HMG. Their melting
temperatures in butter with powder of convective
drying are 1,4 and 0,23 °C lower than melting
temperature of the peaks in control sample. The peak
temperature of LMG and liquid phase of product
decreases for 3,6 °C. On the DSC curves of butter with
powders two peaks of LMG and liquid phase of
ȱ ȱȱ
product are present. The melting temperature of peaks
Figure 1: Curves of butter’s specific heat in comparison with control sample reduced to 2,8 °C
ȱ – fresh-made butter; ȱȱ – after storage ( +5 °ɋ) after adding the powder of CSD. So, there are two
ɚ- control sample; b – butter with carrot powder peaks of MMG and LMG with melting temperatures
of convective drying; c – butter with carrot of 25,9 and 14,5 °C. The melting temperature peak of
powder of cold spray drying.
HMG of 35 °C. It is 2,6 °C more to conformable peak
melting from control. The general trend for both types of butter with the powder compared to the control
is reducing the melting temperature peaks compatible LMG and liquid phase of product. This gives
evidence shows that the powder presence in the system promotes differentiated edging of LMG that are
unable to crystallize during the formation of crystalline phases of butter in the area of lower temperatures.
We believe this is due to the additives’ presence that and their ties in a fatty phase of the product that is
not typical for butter components. During control sample storage the temperature of the melting peaks of
MMG and HMG grow on 2,6 and 1,9 °C. This is due to glyceride’s recrystallization processes. A decline
of melting temperature of LMG and liquid phase of product of butter for 2,8 °C also shows that. The
processes of recrystallization of glycerides’ separate groups undergo during the storage of butter with
carrot powder of convective drying. This is evidenced by the growth of the temperature of the melting
peaks of HMG, NMG and LMG with liquid phase of product. In the butter with powder of CSD the
melting temperature peaks of glycerides individual groups vary slightly after storage. One melting peak of
MMG disappears. That’s why, manifested peaks of melting of LNG and HSV temperatures 14.6 and 35
°C are shown more clearly on the curve.

CONCLUSION
According to the microstructural studies it was revealed that production of carrot powder by cold spray
drying contributes to the formation of particles with a size 1-5 micrometers and saves the powder
component properties. The melting curves of butter showed that the introduction of carrot powder leads to
displacement of fusible glycerides of the solidification front into the zone of lower temperatures. Insertion
of additives of cold spray drying leads to reducing the temperature melting peaks of compatible LMG and
liquid phase product to 2,8 ° C and to increasing the melting temperature peak of HMG to 2,6 ° C in the
fresh butter. During further storage of butter with carrot powder of cold spray drying the temperature
melting peaks of glycerides vary slightly. Adding the powder of convective drying reduces the melting
temperature of compatible LMG and liquid phase product at 4 ° C. During further storage the temperature
melting peaks of glycerides groups gradually increase and are close to the melting temperature of the
control sample.

2024
 Production of ewe’s milk cheese using calf rennet and a plant coagulant from flowers of

cardoon Cynara cardunculus: Proteolysis during ripening

José Fernández-Salguero, Antonio Pino & Elena Galán

Tecnología de los Alimentos. University of Córdoba, Spain (e-mail: ao1fecaj@uco.es)

INTRODUCTION
The crude aqueous extracts from cardoon flowers Cynara cardunculus L. have been traditionally used for
making many cheese varieties mainly in Portugal and Spain, some of which have the Appellation d’Origine
Contrôlée (AOC) status [1, 2]. These acidic enzymes are characterized by high clotting activity like
chymosin, cleaving the peptide bond Phe105-Met106 in bovine and ovine ț-casein, and highly proteolytic in
terms of substrate proteins. This plant coagulant can also be used for producing cheeses aimed at lacto-
vegetarian consumers and ecological markets.
The aim of this work was to study the effect of three coagulants (calf rennet, 100% CR; a powdered
vegetable coagulant, 100% PVC obtained by freeze-drying of crude aqueous extracts from the cardoon
flowers C. cardunculus and a blend of those coagulants, 50% CR/50% PVC) on some chemical and
proteolytic parameters of pasteurized ewe’s milk cheese.

MATERIALS & METHODS

Cheeses from pasteurized sheep´s milk (72ºC/20 s) were obtained using three different coagulants:
powdered vegetable coagulant (PVC), calf rennet (CR) and a blend of both coagulants (CR/PVC). The
coagulation temperature was 29±1ºC. After pressing and salting in brine with NaCl (18%/24 h), the
cheeses (about 3.0 kg weight) were ripened at 12ºC and 80% relative humidity. At 30, 60, 120 and 180
days of ripening were analyzed the components following: moisture, protein, aw, pH, soluble nitrogen
(SN, at pH-4.6), non-protein nitrogen (NPN; 12% TCA), hydrophobic and hydrophilic peptides (RP-
HPLC) and total free amino acids (RP-HPLC) as described elsewhere [3, 4].

RESULTS & DISCUSSION

In moisture, protein, aw, pH (Table 1) and free amino acids (Fig. 1) no differences were observed between
cheeses produced using the three coagulants assayed. The levels of soluble nitrogen and non-protein
nitrogen (Fig. 1) in cheeses produced using plant coagulant were higher than in those produced with calf
rennet (P< 0.05). It was noted that the levels of SN in cheeses made with PVC were very similar to those
obtained using CR/PVC. This coincides with results obtained by O´Mahony et al. [5] in Mini Cheddar
cheeses. These authors explain that although the main agent responsible for the production of SN in
cheese is the residual coagulant, in the case of blend of coagulants are C. Cardunculus proteinases that
dominate the process. So it is possible in sheep's milk cheeses to use a blend of vegetable coagulant from
C. cardunculus and calf rennet to accelerate the ripening. The formation of hydrophobic peptides and the
ratio of hydrophobic/hydrophilic (Fig. 1) peptides (as Area count units x 107) throughout the ripening
were higher (P<0.05) in cheeses obtained with PVC and CR/PVC than in cheeses made with CR.

CONCLUSIONS
The higher proteolytic activity of the cardoon enzymes in contrast to the animal chymosin from the first
stages of ripening could be used in accelerated ripening processes for sheep milk cheese, as the
biochemical characteristics of a ripened cheese are achieved in a shorter amount of time. In consequence,

11th International Congress on Engineering and Food - Athens, Greece, 2011 2025
vegetable coagulant alone or a blend of vegetable coagulant and calf rennet can be feasibly used to
accelerate ripening in sheep milk cheeses.

Table 1. Mean values and standard deviations (±) for moisture and protein (g/100 g cheese), water activity and pH
throughout ripening of the cheeses obtained with CR, PVC and a blend of both CR/PVC
30 60 120 180
aw
CR 0.966±0.004c 0.947±0.011b 0.936±0.005ab 0.930±0.004a
c b ab
CR/PVC 0.970±0.009 0.941±0.019 0.923±0.003 0.923±0.002a
c b ab
PVC 0.972±0.012 0.945±0.014 0.935±0.002 0.925±0.002a
pH
CR 5.21±0.06a 5.25±0.10a 5.22±0.01a 5.25±0.05a
a a a
CR/PVC 5.24±0.02 5.32±0.01 5.25±0.05 5.25±0.02a
a a a
PVC 5.19±0.08 5.24±0.06 5.27±0.11 5.28±0.02a
Moisture
CR 40.6±0.52ef 38.29±0.64de 34.92±0.1bc 33.58±0.88ab
ef de bc
CR/PVC 40.90±1.35 35.91±1.65 35.04±1.05 33.69±2.93ab
ef de bc
PVC 40.36±0.65 40.24±2.39 33.56±1.41 32.88±1.07ab
Protein
CR 21.49±0.02ab 21.91±0.17b 23.86±0.05d 24.55±0.52e
ab b d
CR/PVC 20.53±0.10 21.54±0.07 23.69±0.65 24.59±0.01e
ab b d
PVC 21.21±0.43 21.61±0.22 23.72±0.12 24.65±0.32e
a
Means of the same parameter in the same row without a common superscript letters (a-g) differ significantly (P<0.05)

Figure 1. Average values and standard deviations of the soluble nitrogen (SN) and non-protein nitrogen (NPN), as
g/100 g total nitrogen (TN), total free amino acids (mg/100 g cheese), hydrophilic and hydrophobic peptides and ratio
of hydrophobic/hydrophilic peptides of the cheeses obtained with CR, PVC and a blend of both CR/PVC during the
ripening.

2026
 Production of ewe’s milk cheese using calf rennet and a plant coagulant from flowers of

cardoon Cynara cardunculus: Sensory characteristics during ripening

Elena Galána, Remedios Gonzálezb & José Fernández-Salgueroa

a
Tecnología de los Alimentos. University of Córdoba, Spain (e-mail: ao1fecaj@uco.es)
b
Dep. Psicología. University of Valencia, Spain (e-mail: gonzalrb@uv.es)

INTRODUCTION
The use of plant proteinases from flowers of cardoon Cynara cardunculus as milk coagulants is of
particular interest because they are natural enzymes whose strong proteolytic action eventually leads to
the extensive breakdown of caseins, thereby giving rise to cheeses with a soft buttery texture, a genuine
aroma and a slightly piquant and creamy flavour. These cheeses are highly valued for their taste and
quality and can be targeted at the lacto-vegetarian and organic markets. This type of plant coagulant can
also be certified Kosher and Halal.
This paper reports on a study of the effect of three coagulants: calf rennet (100% CR), a powdered
vegetable coagulant (100% PVC) obtained by freeze-drying of crude aqueous extracts from Cynara
cardunculus L. [1] and a blend of those coagulants (50% CR/50% PVC) on the mainly sensory
characteristics of pasteurized ewe`s milk cheeses during six months of ripening.

MATERIAL & METHODS

The experimental samples of cheese were manufactured as it previously was described and determined
some chemical and biochemical parameters [2]. The cheeses at 60, 90, 120 and 180 days of ripening were
analysed of the attributes following: odour (intensity and acidity), colour of the paste, hardness, firmness,
creaminess, taste intensity and bitter taste, were scored by 12 trained panellists, using a 10-point scale
with anchor points (from “extremely strong” to “extremely mild”) as described elsewhere [3].

RESULTS & DISCUSSION

Table 1 shows the average scores (± standard deviations) awarded by the panel to the various target
attributes of the different cheeses obtained with CR, PVC and CR/PVC during the ripening period.
Cheeses made with animal rennet displayed less odour intensity and acid odour, a slightly lighter colour
and also they were (P<0.05) harder and firmer, but less creamy, than those made with vegetable
coagulant (PVC and CR/PVC). In addition, cheeses obtained using plant coagulant (PVC and CR/PVC)
were greater taste intensity (P<0.05) and a bitter taste slightly higher compared to those made with CR.
Bitter taste of cheese obtained with PVC were higher (P<0.05) compared to those produced using CR but
cheeses obtained using CR/PVC showed intermediate values (P>0.05) compared to cheeses obtained
with CR or PVC. The specificity of C. cardunculus enzymes in ovine casein led to the production of a
normal slight bitter taste in cheeses produced with vegetable coagulant. Therefore, it is possible to
accelerate the ripening of ewe's milk cheese, while still retaining all the organoleptic properties of this
type of cheese, by adding a blend of cardoon C. cardunculus enzymes and calf rennet.

CONCLUSIONS
The use of vegetable coagulant from cardoon flowers of Cynara cardunculus either alone (100% PVC) or
using a blend with calf rennet in equal proportions (50% CR/50% PVC) for sheep cheese manufacture,

11th International Congress on Engineering and Food - Athens, Greece, 2011 2027
results in a cheese with higher scores in taste intensity and creaminess and also with lower scores in
hardness and firmness as opposed to use of calf rennet (CR). In consequence, varying amounts of
vegetable coagulant or blends of vegetable coagulant and calf rennet can be feasibly used to accelerate
ripening in sheep milk cheeses.

Table 1. Average scores (± standard deviations) awarded by the panel to the various target attributes of the different
cheeses obtained with CR, PVC and CR/PVC during the ripening period
Atribute Coagulant Days of ripening
60 90 120 180
Odor CR 5,28±0,52a 6,01±0,44ab 5,51±0,19b 5,61±0,46b
intensity PVC 4,79±0,19a 5,17±0,09ab 6,51±0,21b 6,81±0,04b
a ab b
CR/PVC 4,30±0,10 5,47±0,21 5,64±0,02 5,72±0,12b
a a a
CR 4,74±0,72 4,36±0,13 4,77±0,50 3,96±0,24a
a a a
Acid smell PVC 2,52±1,17 4,85±0,2 4,69±0,06 4,99±1,01a
a a a
CR/PVC 4,39±0,71 4,72±0,06 5,04±0,23 4,49±0,24a
CR 5,52±0,36a 4,86±0,55a 5,88±0,03a 5,67±0,26a
a a a
Color PVC 5,46±0,19 5,94±0,21 6,62±0,00 5,66±0,16a
a a a
CR/PVC 5,21±0,33 5,11±0,59 5,33±0,49 5,83±0,15a
abc bcd cd
CR 5,20±0,37 5,27±0,56 5,95±0,27 6,49±0,44d
a ab a
Hardness PVC 2,81±0,17 3,36±0,21 2,21±0,02 3,38±0,38ab
ab ab ab
CR/PVC 3,66±0,07 3,48±0,16 3,81±0,49 4,90±0,39bcd
CR 5,10±0,23b 5,61±0,13b 6,29±0,03b 6,59±0,36b
a a a
Firmness PVC 2,43±0,01 3,28±0,02 2,10±0,45 2,94±0,34a
a a a
CR/PVC 2,93±0,69 2,93±0,14 3,56±0,21 3,86±10,79a
a a a
CR 6,01±0,55 3,89±0,72 4,36±0,17 3,11±0,06a
Creaminess PVC 7,28±0,26b 7,11±0,13b 7,91±0,34b 7,80±0,31b
CR/PVC 6,02±0,08b 6,92±0,21b 6,87±0,01b 6,40±0,23b
a a ab
CR 5,57±0,73 5,97±0,08 5,61±0,91 6,39±0,21abc
a abc abc
Taste intensity PVC 5,86±0,26 6,28±0,05 6,74±0,19 7,28±0,32c
abc abc abc
CR/PVC 5,27±0,40 6,73±0,35 6,80±0,64 7,10±0,07bc
CR 3,04±0,09a 4,28±0,05a 3,11±0,19a 5,15±1,36a
b b b
Bitter taste PVC 5,27±0,82 5,32±0,29 5,13±0,15 5,38±0,22b
b b b
CR/PVC 4,79±1,02 5,30±0,76 5,41±0,63 5,30±0,45b
a-d
Means of the same row without a common superscript letters, differ significantly (P<0.05).

REFERENCES
[1] Fernández-Salguero, J., Tejada, L. & Gómez, R. (2002). Use of powdered vegetable coagulant in the manufacture
of ewe’s milk cheeses. Journal of the Science of Food and Agricultural, 82, 464-468. [2] Fernández-Salguero J., Pino,
A. & Galán, E. (2011). Production of ewe’s milk cheese using calf rennet and a plant coagulant from flowers of
cardoon Cynara cardunculus: Proteolysis during ripening. 11th International Congress of Engineering and Food.
Athens, Greece, May 22-26, Poster AFT 1242. [3] Tejada,L., Gómez, R. & Fernández-Salguero, J. 2007. Sensory
characteristics of ewe milk cheese made with tree types of coagulant: calf rennet, powdered vegetable coagulant and
crude aqueous extracts from Cynara cardunculus. Journal of Food Quality, 30(1), 91-103.

2028
 Functional Drink Production through Pomegranate Juice Fermentation

S. Plessas*1, M. Koulis1, A. Alexopoulos1 and E. Bezirtzoglou1

1
Democritus University of Thrace, Faculty of Agriculture Development, Laboratory of Microbiology,
Biotechnology and Hygiene, 193 Pantazidou str. GR-68200, Orestiada, Greece, splessas@agro.duth.gr

INTRODUCTION
The strategy of this research survey was to develop a functional drink containing fermented
pomegranate juice with kefir grains. A set of 12 anaerobic fermentation batches of
pomegranate juice were conducted at 3 different temperatures (30, 20 and 10qC) employing
kefir grains. Various analyses were employed in order to evaluate the final product with regard
to microbiological, chemical and organoleptic properties. Concerning the chemical analysis,
the stability of the system was efficient at all studied temperatures, revealing suitability for
industrial applications. Conversion varied between 87.9-92.8%, while ethanol was determined
at respectable amounts, higher than 6%v/v in most cases. Ethanol productivity was determined
in higher levels at the batches conducted at 30qC. GC-MS analysis that conducted for the
pomegranate juice fermented at 10q C showed accepted volatile character due to the
identification of important volatile compounds contributing positively to the aromatic character
of the product. In addition through the determination of cell viability at all the studied
temperatures, possible probiotic properties of the fermented pomegranate juice were revealed
due to the survival of lactic acid bacteria of kefir grains at a concentration of approximately 108
cfu/ml.

MATERIALS & METHODS

Microorganisms
Kefir yeast, a commercial product usually used to produce kefir drink and available at the
Department of Agricultural Development at Democritus University of Thrace, was used.
Anaerobic fermentations
Diluted with sterilized water pomegranate juice (500 ml) and 0.4 g (dry weight) of kefir
biomass and baker’s yeast respectively were dispensed into pre sterilized (with alcohol) conical
flasks glass of 1000ml.. The effect of fermentation temperature (30, 20 and 10°C) on alcohol
production was monitored. Particularly, 5 fermentation batches were conducted at 30°C and
20°C and 2 fermentation batches at 10°C for each microorganism respectively.
Analyses
Ethanol and residual sugar were determined by high performance liquid chromatography,
according to Plessas et al. 2005.
The volatile compounds of the fermented pomegranate juices with kefir and baker’s yeast at
10 °C respectively, were determined by means of gas chromatography–mass spectroscopy
(GC–MS). More specifically, the volatiles were isolated with the headspace solid-phase micro-
extraction (SPME) technique (Plessas et al. 2008).
Counts of lactobacilli in fermented pomegranate juices conducted with kefir grains were
determined as colony forming units (cfu/ml). Results are presented as log of mean colony-
forming units on solid media culture plates containing between 30 and 300 colonies forming
units per ml (cfu/ml) of fermented pomegranate juice.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2029
RESULTS & DISCUSSION
Concerning the chemical analysis, the stability of the system was efficient at all studied
temperatures, revealing suitability for industrial applications as it is represented at Table1.
Conversion varied between 87.9-92.8%, while ethanol was determined at respectable amounts,
higher than 6%v/v in most cases. Ethanol productivity was determined in higher levels at the
batches conducted at 30°C. GC-MS analysis that conducted for the pomegranate juice
fermented at 10° C showed accepted volatile character due to the identification of important
volatile compounds contributing positively to the aromatic character of the product.
Through the determination of cell viability at all the studied temperatures, possible probiotic
properties of the fermented pomegranate juice were revealed due to the survival of lactic acid
bacteria of kefir grains at a concentration of above 108 cfu/ml at the final batch of each
temperature studied. This concentration allows us to characterize the final product as possible
probiotic, since there are many references available in the literature that verifies the probiotic
properties of kefir grains (Garrote et al. 2004; Brialy et al. 1995). However molecular
techniques should be applied in order to identify specific probiotic bacteria.

Table 1. Ethanol production during batch fermentations of pomegranate juice (approximately 100g of
initial sugar concentration) with kefir grains respectively at various temperatures.
Temp. Batch Ferm. Ethanol Residual Ethanol Conversion
time sugar productivity
qC h % v/v g/l g/l/d %
30 1 75 5.9 8.9 14.9 91.1
30 2 74 6.2 7.4 15.9 92.6
30 3 77 6.2 8.1 15.3 91.9
30 4 79 6.2 8.7 14.9 91.3
30 5 82 6.2 8.8 14.3 91.2
20 6 85 6.2 10.2 13.3 89.8
20 7 80 6.2 7.5 14.7 92.5
20 8 81 6.3 8.6 14.7 91.4
20 9 80 6.4 7.2 15.2 92.8
20 10 79 5.1 9.6 12.2 90.4
10 11 204 5.9 10.8 5.5 89.2
10 12 214 5.9 12.1 5.2 87.9

REFERENCES
[1] Plessas S., Bekatorou A., Kanellaki M., Koutinas A. A., Marchant R., & Banat I. M. 2007. Use of Immobilized Cell
Biocatalysts in Baking. Process Biochemistry, 42(8), 1244–1249. [2] Plessas, S., Pherson, L., Bekatorou, A., Nigam,
P., & Koutinas, A A. 2005. Bread Making Using Kefir Grains as Baker’s Yeast. Food Chemistry, 93(4), 585–589. [3]
Plessas, S., Pherson A. Fischer, K. Koureta, C. Psarianos, P. Nigam and A. A. Koutinas 2008. Application of
Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and L. helveticus for sourdough bread making.
Food Chemistry, 106(8), 985-990. [4] Garrote, G.L., Delfederico, L., Bibiloni, R., Abraham, A.G., Perez, P.F.,
Semorile, L. and De Antoni, G.L. 2004. Lactobacilli isolated from kefir grains: evidence of the presence of S-layer
proteins. Journal of Dairy Research, 71, 222-230. [5] Brialy, C., Rivalland, P., Coiffard, L. and de Roeck Holtzhauer,
Y. 1995. Microbiological study of lyophilized dairy kefir. Folia Microbiology, 40,198-200.

2030
Comparative Study of Physico-Chemical Properties and Acceptance Analysis of Different

Formulations of Tapioca Ice Cream

Modesto Antonio Chavesa, Isadora Monteiro Andrade Barretob, Ronielle Cardoso Reisc
a
Universidade Estadual do Sudoeste da Bahia (UESB),Itapetinga, Brazil (modestrochaves@hotmail.com)
b
Universidade Estadual do Sudoeste da Bahia (UESB),Itapetinga, Brazil (isa_mab@hotmail.com)
c
Universidade Federal do Recôncavo Bahiano, Cruz das Almas, Brazil (roniellireis@hotmail.com)

INTRODUCTION
Currently, the ice cream industry has sought creative outlets to entice the consumer among
them one can detach the development of new products. The association of ice cream products
typically linked to regional dishes such as couscous and tapioca meal both typical of north-
eastern Brazil, may be a viable alternative to induce people to consume ice cream throughout
the year, reducing its seasonality[1,2]. The study of physicochemical properties and sensory
related to tapioca ice cream is very important because the results can be used in the formulation
of high quality products that can be accepted by the customers in different markets. This study
had the following aims: - develop an tapioca ice cream testing different formulations, - study
the use of tapioca starch as an agent in tapioca ice cream, - establish the profile of the
consumers of tapioca ice cream in South-western of Bahia State – Brazil, -evaluate acceptance
of different formulations of tapioca ice cream.

MATERIALS & METHODS

The tapioca was prepared to make a porridge using the following ingredients: milk, tapioca,
coconut milk, condensed milk, coconut, and sugar. The tapioca porridges were prepared by
adding two concentrations of tapioca (6 and 10% w/w) and two cooking temperatures (40 and
60oC). The tapioca was added in two distinct steps of processing (1 - before maturity, 2 - after
maturation. The tapioca ice cream formulations were characterized by performing the
following physical-chemical properties analysis: colour, total solids, moisture (percentage
w.b.), soluble solids (°Brix), pH and acidity, viscosity at 7ºC, overrun, melting test, and
apparent density. The tapioca ice cream was subjected to an acceptance test in which the
sensory attributes of colour, aroma, flavour, and consistency were evaluated by 81 panellists
answering a hedonic scale of nine points. Both data on the physical and chemical properties
and the results of the acceptance test were submitted to analysis of variance at 5% probability
and mean tests.

RESULTS & DISCUSSION

Table 1 presents the average values of physico-chemical properties of the eight different
formulations of tapioca ice cream. The analysis of variance (ANOVA) showed that the
treatments did not differ at 5% significance, by the F test for differences in the physico-

11th International Congress on Engineering and Food - Athens, Greece, 2011 2031
chemical properties evaluated. It was observed that the pH ranged from 6.35 to 6.63 is in
agreement with the values found by Schmidt et al[3]. The formulation containing 6% of
tapioca (S62) had the highest overrun (49%). It is observed that the viscosities in the
formulations containing 6% of tapioca are smaller than those in the formulations with 10%.
With regard to the habits and frequency of consumption of, it can be sad that the consumption
of ice cream and tapioca were extremely high (100 and 99% respectively) in the region.
Among the consumers, 62% had already eaten tapioca ice cream and 38% of consumers had
never consumed.

Table 1. Average values of the physico-chemical properties of tapioca ice cream for all treatments.

S41: 6% of tapioca/40°C/1th Step; S42: 6% of tapioca/40°C/2nd Step; S61: 6% of tapioca/60°C/1th Step; S62: 6% of
tapioca/60°C/2nd Step; D41: 10% of tapioca/40°C/1th Step; D42: 10% of tapioca/40°C/2nd Step; D61: 10% of
tapioca/60°C/1th Step; D62: 10% of tapioca/60°C/2nd Step.

Regarding the factors considered most important by consumers when choosing and buying the
product the attribute considered most important was the taste (31.8%), followed by consistency
(20.9%) and price (20.2%). Combining all the attributes evaluated the best tapioca ice cream
was formulated with 6% of tapioca cooked at 60°C and added before maturation (Step1).

CONCLUSION
Concerning the acidity, pH, moisture content, total solids, soluble solids and colour, the tapioca
ice cream formulations used herein, are equivalent. All formulations of tapioca ice cream had
good acceptance. However, the formulation with 6% of tapioca, cooked at 60 ° C and added
before maturation, was the most acceptable.

REFERENCES
[1] Clark, K.B & Wheelwright, S.C. 1993.Managing new product and process development: text and
cases. New York: The Free Press, 135p
[2] Porter, M. 1989. Vantagem competitiva: criando e sustentando um desempenho superior. Rio de
Janeiro: Editora Campus.
[3] Schmidt, K.; Lundy, A.; Reynolds, J.; Yee, L.N. 1999. Carbohydrate or protein based fat mimicker
effects on ice milk properties. Journal of Food Science,58(4)761-799.

2032
 Bread-making potential of pea protein isolate produced by a novel

ultrafiltration/diafiltration process

Louis-Philippe Des Marchais, Mathieu Foisy, Samuel Mercier, Sébastien Villeneuve, Martin Mondor

Food Research and Development Centre, Agriculture and Agri-Food Canada, 3600 Casavant Blvd West,
Saint-Hyacinthe, Quebec, Canada, J2S 8E3 (sebastien.villeneuve@agr.gc.ca)

INTRODUCTION
The incorporation of ingredients like legume flour, concentrate or isolate in cereal-based
matrices can lead to the production of nutritionally enhanced products like bread with high
protein content. However, many ingredients currently available on the market have a large
phytate to protein ratio resulting in reduced protein digestibility and minerals bioavailability.
Moreover, substitution of wheat flour with legume-based ingredients at a 10% level or more is
generally harmful to the processing of bread. This work aimed to study the potential of
supplementing bread with pea protein isolate having a low phytate to protein ratio produced by
a novel ultrafiltration/diafiltration (UF/DF) process.

MATERIALS & METHODS

Pea protein isolate produced by membrane technologies were made from certified #1 Eclipse
Yellow peas (Churchbridge, SK, Canada). Isolate were produced by extracting pea flour in
water at room temperature (ratio 1:15 w/w) and pH 7.5 followed by purification with UF/DF
using 50 kDa hollow fibres membranes. The UF/DF sequence was a UF step with a volume
concentration ratio (VCR) 5 and a discontinuous DF step with a re-VCR 5. The resulting pea
protein isolate was lyophilized and placed in aluminium pouches which were hermetically
sealed and stored at 4°C until used [1]. Commercial wheat flour (Horizon Milling, Montreal,
QC, Canada) was then substituted at a 10% level (dry basis) with isolate in order to produce
bread with protein content over 20%. Both commercial and substituted flour properties, dough
mixing properties and bread characteristics were measured according to AACC International
methods (Damaged Starch, Falling Number, Farinograph Method for Flour, Optimized
Straight-Dough Bread-Baking Method). Dough properties were also characterised using a
modified version of the continuous water addition method developed by Landillon, et al. [2].
Three parameters were determined: minimum water content for dough formation, maximum
torque and water content at maximum torque. Color measurements of crust and crumb were
performed using a colorimeter and expressed in terms of L*, a* and b* parameters.

RESULTS & DISCUSSION

The protein content of the pea protein isolate was 96.1±0.2% (dry basis) resulting in dough
protein content of about 23% (dry basis). Supplementing wheat flour with pea protein isolate
did not affect damaged starch but slightly decreased falling number (Table 1). Water
absorption was increased for substituted flour while dough stability and development time were
not affected. The addition of isolate caused a diminution of the minimum water content for
dough formation under continuous water addition. Furthermore, higher maximum torque was
observed for the substituted flour with corresponding lower water content at maximum torque.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2033
Substitution with pea protein isolate allowed to produce bread with protein content over 20%
(20.9±0.1% compared to 14.5±0.0% dry basis) but induced a decrease in loaf specific volume
(4.3±0.0 cm3 g-1 compared to 5.4±0.3 cm3 g-1) compared to unsubstituted flour. A protein
content of this magnitude is relatively high considering that reports in the literature conclude
that high protein breads contain up to 15-20% protein [3]. Moreover, the specific volume of
bread substituted with pea protein was maintained over 4 g cm-3, which should be considered
as satisfactory by the consumers. The substitution of wheat flour with isolate induced a
decrease in the whiteness and an increase in the yellowness of the crumb, while inducing an
increase in the blackness and a decrease in the yellowness of the crust.

Table 1. Protein content, absorption of Iodine, falling number and mixing properties of dough
Dough
Dough with
enriched with
Parameters 100% flour
10% pea
(control)
protein isolate
Moisture content (% dry matter) 13.5 ± 0.1 12.7 ± 0.1 ***
Protein content (% dry matter) 15.2 ± 0.2 23.3 ± 0.2 ***
Absorption of Iodine (%) * 94.85 ± 0.01a 94.82 ± 0.03a
Falling number (s) ** 358 ± 4a 344 ± 3b
Water absorption (%) ** 59.3 ± 0.1a 62.5 ± 0.1b
Dough development time (min) ** 6 ± 1a 7 ± 1a
Stability (min) ** 6.8 ± 0.9a 8.0 ± 0.7a
Maximum torque (FU) * 867 ± 75a 965 ± 18b
-1
Minimum water content for dough formation (mL g-solid ) ** 0.46 ± 0.01a 0.41 ± 0.00b
Water content at maximum torque (mL g-solid-1) ** 0.61 ± 0.02a 0.54 ± 0.01b
*Values not sharing a common letter are significantly different (P < 0.05); **Values not sharing a
common letter are significantly different (P < 0.01); ***Calculated values

CONCLUSION
Flour substituted with low phytate pea protein isolate produced by UF/DF showed a good
bread-making potential when compared to unsubstituted flour. Next step will be to assess the
potential improvement in protein digestibility and mineral bioavailability for the bread
substituted with this new pea protein isolate compared to other alternative ingredients. This
could lead to the marketing of high protein breads with improved health properties.

REFERENCES
[1] Taherian, A.R., Mondor M., Labranche J., Drolet H., Ippersiel D. & Lamarche F. 2011. Comparative
study of functional properties of commercial and membrane processed yellow pea protein isolates.
Food Research International, doi:10.1016/j.foodres.2011.01.030 (in press).
[2] Landillon V., Cassan D., Morel M.H. & Cuq B. 2008. Flowability, cohesive, and granulation
properties of wheat powders. Journal of Food Engineering, 86,178-193.
[3] Mohamed A.A., Rayas-Duarte P., Shogren R.L., & Sessa D.J. 2006. Low carbohydrates bread:
formulation, processing and sensory quality. Food Chemistry, 99, 686-692.

2034
 Technology of functional public catering foods with dietary additives

K.V.Svidloa, M.I.Peresichnyib

a
Department of Trade, Hotel and Restaurant and Tourism Industry, Kharkiv Institute of Trade
and Economy of Kyiv University of Trade and Economy,Ukraine (karinasvidlo@rambler.ru)
b
Department of Trade, Hotel and Restaurant and Tourism Industry, Kyiv University of Trade
and Economy,Ukraine (frh@knteu.kiev.ua)

INTRODUCTION
The technology of making functional foods is based on both traditional ingredient
modifications and the ones with useful ingredients extended to the level comparable with
physiological consumption rates within the range of 10-50% ( according to various sources) of
the average daily consumption. Lately, Ukrainian functional food producers have been scaling
up the use of vegetable dietary additives . The ministry of public health of Ukraine permits the
use of ground vegetable seeds as dietary additives.
Extensive research carried out in many countries shows that one of the main causes of
pathologic processes in human organism, which provoke numerous diseases and early aging,
arise from extensive accumulation of free radicals. Their ruinous effect is safely protected by
ground vegetable seeds which contain biological antioxidants. They are natural non-toxic
phytogenous compounds that neutralize free radicals. Russian and American scientists [1-2] in
their study of using ground vegetable seeds in bakery, macaroni and confectionary
technologies have commonly acknowledged positive effects of ground flax, holy thistle and
sea-buckthorn seeds on functional and technological properties of foodstuffs, their nutritive and
biological value. They approved of the possibilities of using ground vegetable seeds for
efficient public catering and disease prevention.

MATERIALS & METHODS

To study the character of changes of gluten properties by adding ground seeds into wheat flour
the research was carried out on model samples with varying content of ground seeds. Wheat
flour of equal quality levels was used for the samples with various kinds of ground seeds.

RESULTS & DISCUSSION

The results of the research show that the amount of washed off gluten decreases with increase
of ground flax, pumpkin and holy thistle seeds. At the same time, in the case of increasing
ground flax seed content the amount of gluten decreases at a higher rate than in the case when
we use ground pumpkin and holy thistle seeds. The lower content of washed off gluten as
compared to the check can be explained by seeds having a high content of biologically active
components of lipidic nature and nutritive fibers, mainly cellulose. When interacting with
wheat flour cellulose they are likely to worsen their ability to create bound gluten paste. This is
confirmed by their decrease in quantity. On adding ground oat seeds, the quantity of washed
off gluten increases within the range of 0.5- 6% of the wheat flour weight. Further, decrease of
washed off gluten in comparison with the check is observed. This is likely to be related with

11th International Congress on Engineering and Food - Athens, Greece, 2011 2035
the fact that aleurone oat layer and its shell contain much cellulose, hemi-cellulose (about 55%)
and lignin which create a system with high sorption properties.
The results show that ground flax seeds, when swell, hold water one and a half times as much
as ground holy thistle seeds and three times as much as ground pumpkin seeds do (Table 1).
This is likely to hinder the sufficient swelling of wheat flour and formation of bound gluten.

Table 1. Study of the process of ground seed swelling against time and hydromodulus
Type of ground seeds Hydro Swell percentage against time, (min.)
modulus 5 15 20 30 45 60
Ground pumpkin seeds 1:5 205 223 250 254 255 258
Ground pumpkin seeds 1:10 220 237 256 259 266 268
Ground holy thistle seeds 1:5 375 382 389 395 397 400
Ground holy thistle seeds 1:10 421 433 437 440 443 448
Ground flax seeds 1:5 503 573 588 600 612 624
Ground flax seeds 1:10 710 754 776 789 803 817

Our research also indicates that adding ground seeds not only decreases washed off
gluten quantitatively, but deteriorates it qualitatively. Increased quantity of ground pumpkin,
flax and holy thistle seeds leads to lower quality of gluten, but adding ground oat seeds within
the range of 0.5-6% of the wheat flout weight improves it. Therefore, some seed additives
weaken the ability of gluten to create firm spatial structure. The research results show that
ground flax seeds have far greater influence on gluten quality properties than in the case with
oat seeds. Gluten quality factors (extensibility, elasticity) for ground pumpkin and pumpkin-
with-spirulina seeds are average for gluten properties of ground flux and oat seeds. This is due
to the fact that oats contain phytin acids which tend to make wheat pastry gluten stronger.
Flux seeds contain specific polysaccharides (gum) which affect function-technological
properties of wheat pastry. Their content amounts to 9-12% of the weight of solid seeds.

CONCLUSION
It is evident that in order to make high quality yeast and sheet pastry with sound wheat flour,
additives are introduced into the pastry to improve gluten elasticity. Therefore, low amounts of
ground oat seeds can be used in yeast and sheet pastry from sound flour. Ground flax, pumpkin
and holy thistle seeds are advisable as additives for plastic-type pastry, particularly for shot
pastry. Further investigation in this field requires extended studies of effects of replacing wheat
flour on ground seed additives by using concrete food systems and taking into account specific
culinary and confectionary technology peculiarities.

REFERENCES
[1] Manthey F.A., Lee R.E., Hall C.A. Processing and cooking effect on lipid content and stability of
alfa-linolenic acid in spaghetti containing ground flaxseed // J. agr. Food Chem. -2002.-Vol. 50.-N.6.
-P.1668-1671.
[2] ɂɫɩɨɥɶɡɨɜɚɧɢɟ ɫɟɦɹɧ ɥɶɧɚ ɜ ɯɥɟɛɨɩɟɱɟɧɢɢ/ ɂ.Ɇɢɧɟɜɢɱ, ȼ.Ɂɭɛɰɨɜ, Ɍ.ɐɵɝɚɧɨɜɚ//
ɏɥɟɛɨɩɪɨɞɭɤɬɵ. -2008. -ʋ3.- ɋ.38-40.

2036
 Production of chromium-chelating peptides after hydrolysis

of silk fibroin protein with elastase

Hu Changlia Chen Lijuna* Ren Fazhengb

a
Beijing Sanyuan Foods Co. Ltd., Technique CenterˈBeijing 100085, China
(hu_changli@126.com)
b
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing100083,
China (renfazheng@263.net)

INTRODUCTION
The beneficial effects of the element on glucose and lipid metabolism have prompted
investigations into the importance of Cr supplementation in the human diet. In the recent years,
food intake is increasingly being considered not only as a source of nutrients but also as a
source of bioactive compounds, including bioactive peptides.
Silk is composed of two kinds of proteins: fibroinand and sericin. These proteins may present
an interesting raw material for the production of protein hydrolysates enriched in bioactive
peptides. SF may have strong affinity for several elements through the chelation of their
hydroxyl and carboxyl groups to the elements. The positive effect of Silk sericin peptides on
the in vivo and in vitro absorption of minerals such as Zn, Fe, Mg and Ca have been reported
[1]
. The production and purification of chelating peptides during digestion of SF protein isolates
with elastase and the function of Cr-chelating peptides was described in the present paper.

MATERIALS & METHODS

Preparation of SF protein hydrolysastes
Enzymatic degradation was carried out in a reactor using the pH-stat method. Elastase was
added into the sample at a concentration and terminated by heating the solution to 100ć for 15
min. The supernatant collected for further experiments.
Determination of the Degree of hydrolysis (DH)
The degree of hydrolysis was calculated by determination of free amino groups by reaction
with TNBS.
Assay for generation of hydroxyl radicals
The potential of Cr compounds to generate hydroxyl radicals in vitro was assessed by the
method of Gutteridge et al. [2]. And the amount of chromogen formed in the sample was
measured by its absorption at 520 nm. Ferric-EDTA (100 ȝM) was used as a positive control.
The Sephadex-G-25 Gel Chromatography
Purified Cr-chelating peptides were loaded into a Chromatography column (800×20 mm). The
speed of elution was set at about 0.5 mL/min. All the samples were inspected with HD-3 type
spectrophotometer under the wavelength 280 nm at the same time automatically.
The mid infrared (MIR) Spectroscopy analysis
The construction of the sample was analysis by MIR Detector (ANTARIS, Thermo nicolet
USA) at the range of 4000 - 400 cm-1.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2037
RESULTS & DISCUSSION
Function of Cr-chelating SF protein hydrolysates
The elastase activity is observed even after 10 h of incubation with the SF protein. Protein
hydrolysates obtained at different times were indirectly assayed for their Cr-chelating capacity
by determination their effect on DNA damage. All Cr-chelating SF peptides inhibited to certain
degree of scavenging effect on free radical, but in general hydrolysates obtained by the action
of elastase were more effective at a long incubation time than a short time. Thus, most
scavenging hydrolysate was obtained after 480 min incubation with elastase.
Purification of Cr-chelating peptides fractions
In this experiment, the Sephadex G-25 was used as phase vector to separate the Cr-chelating
SF peptides. Eluted peptides were fractionated in four fractions that were assayed for
generation of hydroxyl radicals (Figure 1). The smaller molecular, eluted later, were of the
most scavenging activities, while the bigger molecular, eluted first, were of less scavenging
activities such as fraction 4.




OD(520 nm)








   
Fractions collected from Sephadex G-25

Figure 1. The scavenging activities of fraction 1-4 collected.

The structure analysis of the sample by MIR Detector

The MIR spectra of SF hydrolysates and Cr-chelating peptides were different evidently.
Compared to SF hydrolysates, changes in this region (3,277.31 cm-1) can be observed for Cr-
chelating SF peptides.

CONCLUSION
The protein hydrolysates obtained after 480 min incubation with elastase was the most
scavenging free hydroxyl radicals. Thus, food proteins may be responsible of improving metal
bioavailability after protein hydrolysis and release of peptides with chelating properties as
those obtained in our in vitro hydrolysis of SF protein with elastase.

REFERENCES
[1] Masahiro S., Hideyuki Y. and Norihisa., 2000. Consumption of Silk protein, sericin elevates intestinal
absorption of Zinc, Iron, Magnesium and Calcium in rats. Nutrition Research, 20(10), 1505-1511.
[2] Halliwell B., Gutteridge J.M. and Aruoma O.I., 1987. The deoxyribose method: a simple test-tube
assay for determination of rate constants for reactions of hydroxyl radicals. Anal. Biochem, 165, 215-
219.

2038
11th International Congress on Engineering and Food - Athens, Greece, 2011 2039
plants. Only freeze-dried lemon balm exhibited higher content of polyphenols when compared to
fresh plant.

120.00 250.00

TPC ABTS
100.00
200.00

80.00
mg GAE/g DM

umol Trolox/g DM
150.00

60.00

100.00
40.00

50.00
20.00

0.00 0.00
Fresh AD FD MD Fresh AD FD MD Fresh AD FD MD

Lemon balm Borage Marigold

Figure 1. Total phenol content (TPC) and antioxidant capacity of plant extracts
determined using the ABTS assay

The content of carotenoids decreased after drying, when compared to fresh plants, which indicates
the susceptibility of these beneficial bioactive compounds to degradation at higher temperatures.
Although it could be expected that fresh plants contain the highest chlorophylls content, the highest
content of chlorophylls was determined in freeze-dried plants.

CONCLUSIONS
The antioxidant capacity of all evaluated bioactive compounds was in agreement with the content
of the attributing compounds, thus confirming the high bioactive potential of lemon balm, marigold
and borage. Fresh plants were characterized with the highest contents of carotenoids and
polpyhenols, the highest content of chlorophylls was determined in freeze-dried plants, while air-
dried plants exhibited the highest content of anthocyanins. Microwave drying resulted with a
significant degradation of all bioactive compounds in the examined plants.

REFERENCES
[1] Garg H.P. & Kumar R. 2001. Developments in solar drying, In: Proceedings of the Second Asian-Oceania
Drying Conference (ADC 2001), Batu Feringhi, Pulau Pinang, Malaysia, 297–319.
[2] Jambor J. & Czosnowska E. 2002. Herbal medicines from fresh plants. PostĊpy Fitoterapii, 8(1–2), 2–5.
[3] Diplock T.A., Charleux J.L., Crozier-Willi G., Kok F. J., Rice-Evans C., Roberfroid M., Stahl W. & ViĖa-
Ribes J. 1998. Functional food science and defence against reactive oxidative species. British Journal of
Nutrition, 80, 77–112.

2040
 Research on Dehydrated Fruit Leathers: A Review

Natalia A. Quintero Ruiza, Silvana M. Demarchia, Sergio A. Ginera,b,c

a
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), La Plata, Argentina
b
Comisión de Investigaciones Científicas (CIC), La Plata, Argentina (saginer@ing.unlp.edu.ar)
c
Facultad de Ingeniería, Universidad Nacional de La Plata, La Plata, Argentina

INTRODUCTION
Fruit leathers are dehydrated fruit-based products that are eaten as candy or snacks, and
presented as flexible stripes or sheets. They receive this name because of the final product
aspect (it is shiny and has the texture of leather). The origin of fruit leathers may go back to the
Persian Empire. They are known as “Pestil” in Turkey, “Bastegh” or “Pastegh” in Armenia,
“Qamar al deen” in Lebanon, Syria and other arab countries and “Fruit roll” or “Fruit leather”
in the United States. The last denomination is possibly more usual in the scientific literature
[1]. Due to its novel and attractive structure, and for being products that do not require
refrigeration, they constitute a practical way to incorporate fruit solids, especially for children
and adolescents. Fruit leathers allow leftover ripe fruits to be preserved. In recent years, their
popularity has increased, transforming from a homemade preparation into an industrial
product. The available literature describes two main methods for preparing fruit leather
structure: pectic gelation of fruit puree or concentrates induced by dehydration and non-pectic
gelation, using starch or other gelling agents. This review will consider research works on
leathers arrived at by pectic gelation.

PREPARATION OF FRUIT LEATHERS

The process of pectic gelification leading to a fruit leather has the following requirements: a
soluble solid content greater than 55% w/w, composed of fruit pulp and, optionally, by added
saccharides. Besides, the pH of the formulation must be of 3.5 or below. Pectins with high
degree of esterification are necessary as well [2]. A general flow chart for the production of
fruit leathers is shown in Figure 1. This process may vary according to the fruit used, the nature
of additives that may be employed and the drying technology.

Figure 1. Flow-sheet for preparation of fruit leather

A BRIEF LITERATURE REVIEW OF RESEARCH ON FRUIT LEATHERS

The first contributions (1976) describing the preparation of fruit leathers were written by
extension services of various universities in the United States [3]. These techniques were aimed

11th International Congress on Engineering and Food - Athens, Greece, 2011 2041
at promoting homemade preparation of leathers from several fruits. Scientific production in the
topic began around 1978 and, despite the healthy character of fruit solids consumption, has
kept an irregular pace until the beginning of the XXI century, from which fruit leathers began
to receive more attention from researchers. Early work described the physicochemical
properties and sensorial attributes of pectic gels. Subsequent works incorporated data on
quality parameters and storage stability [4]. In the last decade, publications included subject as
the use of combined technologies [5], analysis of drying kinetics, application of the glass
transition temperature (Tg) theory to fruit leathers and evaluations of the influence of various
additives in drying rate and product quality. Latest research on fruit leathers aimed at studying
the effects of processing on organoleptic and nutritional quality of the final product.

CONCLUSIONS
Fresh fruits and vegetables are known to be excellent sources of energy, minerals, vitamins and
bioactive compounds (phenolics, carotenoids) and fiber. The destruction of the original fruit
structure by pureeing and its reestructuring in dehydrated sugar-acid-pectic gels called fruit
leathers provides attractive, colored products, on which research is very active nowadays. This
trend towards quality analysis was observed in research carried out in recent years, especially
as affected by different process and storage conditions. Concerning organoleptic quality, color
is usually selected as quality parameter, because it has a high impact on consumers and is
useful as browning index. Nutritional quality may be considered as an important parameter
when processing fruits that are source of vitamins and/or antioxidants. Future research should
address the important issue of the effect of processing on quality, not only to define process
conditions in a given technology, but also to compare diverse technologies. This knowledge is
required by industry as well as by consumers. Conventional hot-air drying processes, still the
more common technology, produces strong antioxidant loss, though increase the availability of
a microbiological stable product. Quality losses progress during storage at ambient
temperatures and the “functional” character of these products may be preserved only if dried
with modern hybrid or combined technologies, mostly under vacuum and stored at
refrigeration temperatures to slow down antioxidant loss. There is still not a unified criterion
about the physicochemical characteristics representing a fruit leather, so that a definition of its
main composition or preparation method should be defined, or standardized to inform
consumers and to favor trade at national and international level.

REFERENCES
[1] Maskan A., Kaya S. & Maskan M. 2002. Hot Air and Sun Drying of Grape Leather (Pestil). Journal
of Food Engineering, 54, 81-88.
[2] Visser J. & Voragen A. 1995. Pectins and Pectinases. Elsevier Science B.V., Amsterdan, Holand.
[3] Raab C. & Oehler N. 1976. Making Dried Fruit Leather, Oregon State University. Extension Service,
Oregon, USA.
[4] Vijayanand P., Yadav A. R., Balasubramanyan N. & Narasimham P. 2000. Storage Stability of Guava
Fruit Bar Prepared Using a New Process. Lebensmittel-Wissenschaft und-Technologie, 33(2), 132-
137
[5] Drouzas A. E., Tsami E. & Saravacos G. D. 1999. Microwave/Vacuum Drying of Model Fruit Gels.
Journal of Food Engineering, 39(2), 117-122.

2042
 Selection of potential probiotic Lactobacillus strains from human milk

Hatice Yavuzdurmaza, Sebnem Harsa a

a
Izmir Institute of Technology, Izmir, Turkey (haticeyavuzdurmaz@iyte.edu.tr)

INTRODUCTION
Probiotics are suggested as food to provide for the balance of intestinal flora. Probiotics have
been used for long time in food ingredients for human and also to feed animals without any
side effects. The human breast milk has been considered to be an attractive source for potential
probiotic strains. After birth, breast milk becomes the best food for infants because it fulfills all
required nutrients. Based on the microbiological point, human milk is really an important
factor in the initiation and development and of course, for the composition of the neonatal gut
microflora since it constitutes a source of microorganisms to the infant gut for several weeks
after birth. Although there are limited knowledge about the commensal or probiotic bacteria in
breast milk, bacteria commonly isolated from this biological fluid include staphylococci,
streptococci, micrococci, lactobacilli and enterococci. The main scope of this study is to isolate
and identify lactobacilli and search the potential probiotic properties of these isolates.

MATERIALS & METHODS

Samples were collected from healthy mothers in sterile carriers. Pour plate technique was used
to isolate the organisms. Serial dilutions were plated onto Man, Rogosa and Sharp (MRS) agar
(pH 6.2 and 5.5), TPY (Trypticase Phytone Yeast) agar (pH 6.5) and MRS-cystein agar (pH
5.5). Plates were incubated anaerobically at 37 °C for 72 h. Gram-positive and catalase-
negative rods and coccoid shaped ones were randomly selected. Isolated strains were analyzed
based on their resistance to low pH (3.0), tolerance against bile (0.3% bile salt) and
antimicrobial activity. Then, isolates showed potential probiotic characteristics were
characterized phenotypically and identified genetically by using 16S rDNA sequencing. The
biochemical characteristics were determined by analyzing: CO2 production from glucose,
growth at different temperatures (10, 15, 45 °C), growth at different NaCl concentrations (2%,
6.5%), hydrolysis of arginine and fermentation ability of 17 different carbohydrates. After
determination of biochemical characteristics genomic DNA was isolated according to the
protocol of which is modified from the protocol of Cardinal et al [1]. Amplification of 16S
rDNA region was performed with EGE1 forward primer (5’-AGAGTTTGATCCTGGCTCAG-
3’) and EGE2 reverse primer (5’-CTACGGCTACCTTGTTACGA-3’) [2]. The amplification
conditions were as follows: 94°C for 5 min (initial denaturation); 40 cycles of 94°C for 1 min
(denaturation), 56°C for 1 min (annealing), 72°C for 1 min (elongation); 72°C for 10 min (final
extension). The amplification conditions were as follows: 94°C for 5 min (initial denaturation);
40 cycles of 94°C for 1 min (denaturation), 56°C for 1 min (annealing), 72°C for 1 min
(elongation); 72°C for 10 min (final extension). Then PCR products were sequenced and
analyzed by using the basic local alignment search tool (BLAST,
http://blast.ncbi.nlm.nih.gov/).

RESULTS & DISCUSSION

11th International Congress on Engineering and Food - Athens, Greece, 2011 2043
From 200 isolates, 60 isolates remained at the end of the isolation, purification after the loss of
unstable isolates during purification and subculturing steps. All of the isolates were gram
positive catalase negative rods and cocci. In order to select the most resistant strains to low pH
values, PBS buffer adjusted to pH 3.0 was used and cfu (colony forming unit) values were
obtained during 3 h. Only two bacilli isolates survived in pH 3.0 and then these isolates were
screened for their ability to tolerate the bile salt. Strains were detected in 0.3% bile during 4 h.
The cfu values showed that both isolates were resistant to bile at this concentration (Figure1).
Afterwards, antimicrobial activity tests were assayed towards Salmonella typhimurium CCM
5445, Escherichia coli O157:H7 NCTC 129000 and Escherichia coli NRRL B-3008. The
diameter of inhibition zones indicated that isolates had antibacterial effect on the indicator
microorganisms. Both isolates showed much more efficiency on Escherichia coli NRRL B-
3008. After searching probiotic properties, isolates were characterized by physiological and
biochemical methods. When biochemical experiment findings were compared with the
literature information, it was provided that AS17 was like to be Lactobacillus oris; AS83 was
like to be Lactobacillus fermentum. Ultimately, these two strains were subjected to 16S rDNA
sequencing and identified as Lactobacillus oris (AS17) and Lactobacillus fermentum (AS83).

Figure 1. Survival in pH 3.0 and Tolerance against 0.3% bile

CONCLUSION
These two strains identified as Lactobacillus fermentum and Lactobacillus oris have the
potential to be used as probiotic strains. They both survive at low pH, resistant to bile salt and
have a considerable antimicrobial effect against the tested bacteria. Our results showed
similarity on the base of isolation of Lactobacillus fermentum from human breast milk with the
other isolated strains from this liquid. However this is the first study for the isolation of
Lactobacillus oris from human breast milk. Therefore, Lactobacillus oris is might be used as
commercial probiotic because it provides the prerequisites for being probiotic. Human milk is
also a potential natural source may be used to isolate probiotic lactic acid bacteria.

REFERENCES
[1] Cardinal M. J, Meghrous J., Lacroix C. & Simard R. E. 1997. Isolation of Lactococcus lactis strain
producing inhibitory activity against Listeria. Food Biotechnology, 11, 129-146.
[2] Mora D., Fortina M. G., Nicastro G., Parini C. & Manachini P. L. 1998. Genotypic characterization of
thermophilic bacilli: a study on new soil isolates and several reference strains. Research in
Microbiology, 149, 711-722.

2044
 Phenolics, betalains, ascorbic acid, and antioxidant activity of Opuntia ficus-indica.

Dulce María Jiménez-Aguilara. Cármen Hernández-Brenesb. Janet Alejandra Gutierrez-Uribec. Jorge Welti-Chanesd.
a,b,c,d
Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Av. Eugenio Garza Sada
2501 Sur, CP 64849. Monterrey, NL, México. aA00808278@itesm.mx, bchbrenes@itesm.mx, cjagu@itesm.mx,
d
jwelti@itesm.mx.

INTRODUCTION
Prickly pear is one of the most representative fruits in Mexican culture, it is a fruit which presents a
thick pericarp with small prickles, enclosing a pulp, which is intermixed with a number of small
seeds. Prickly pear fruit is an important source of sugars, minerals, aminoacids, phenolic
compounds, betalains, and vitamin C. Phenolic compounds have anti-inflammatory, anti-allergenic,
anti-inflammatory, anti-atherogenic, and cardioprotective effects, and these are potential
antioxidants.
In Puebla, approximately 41,672 ton/year of prickly pear are harvested, making this State one of the
main producers of this fruit in México [1]. However, commercial varieties obtained from this
location have not been studied. The objective of this study was to quantify the content of total
phenolics, total betalains, ascorbic acid and antioxidant activity found in the juice, pulp and rind of
the two commercial varieties of prickly pear originating from Puebla, Mexico.

MATERIALS & METHODS

Biological material and sample preparation. Two commercial types of Prickly pear (Opuntia
ficus-indica) denominated Red San Martín and Green Villanueva, were harvested during 2010, in
February and May, respectively. Harvesting took place in San Sebastián Villanueva, Puebla,
México. The prickly pear pulp was separated from the rind and seeds. The juice was obtained from
the pulp by paper filtration. Rind was freeze-dried, and then was crushed for 3 minutes. 1 g of the
dried rind samples was homogenized with 10 mL of methanol (80%). The homogeneized products
were mixed during 2.5 h at 250 rpm and 25 ± 0.5 °C (Lab Line model 3526, USA). Afterwards,
samples were centrifuged at 5000 g and 4°C for 10 min (IEC centrifuge model MP4R, USA). Rind
extract, juice and pulp were stored at -80 °C
Total phenolics, total betalains, and ascorbic acid. Total phenolic content was evaluated
according to the Folin Ciocalteau method described by Stintzing et al [2]. Total betalains content
was measured according to Cassano et al. [3]. The extraction and measurement of ascorbic acid
were performed as described by Gillespie and Ainsworth [4].
Antioxidant activity. Antioxidant activity was evaluated with the method of Oxygen Radical
Absorbance Capacity (ORAC) described by Huang et al [5].

RESULTS & DISCUSSION

Total phenolics. Table 1, summarizes the composition of TP in juice, pulp and rind of the
Villanueva and San Martín prickly pears. The juice of San Martín prickly pear had higher levels of
phenolics than Villanueva.
Betalains. The juice and the pulp of the San Martin prickly pear (Table 1) showed higher
concentrations of betacyanins than of betaxanthins. The rind of Villanueva prickly pear showed
lower levels of betalains, meanwhile in the juice and the pulp they could not be detected. Ascorbic

11th International Congress on Engineering and Food - Athens, Greece, 2011 2045
acid (AA). The juice of San Martin prickly pear had the highest levels of AA. In the juice and pulp
of Villanueva, it was found 2.5 times less AA than in San Martin juice. The rind had the lowest
concentrations of AA (Table 1).
Antioxidant activity (AOX). In Table 1, it can be observed that both varieties of prickly pear had
higher levels of antioxidant activity in the rind. Juices and pulps studied showed similar levels;
nevertheless the AOX in the rind of Villanueva was higher.

Table 1. Total phenolics, betalains, vitamin C and antioxidant activity in juice, pulp and rind of
commercial varieties of prickly pear harvested in Puebla, México
Variety Total phenolics Betalains (mg/100 g) Vitamin C Antioxidant
activity
(mg GAE/100 Betacyanins Betaxanthins (mg AA/100
g) g) (ȝmol TE/g)
San Martín Juice 68.3 ± 1.3a 26.8 ± 0.5a 17.7 ± 0.4a 58.2 ± 0.6a 9.8 ± 0.5a
Villanueva Juice 53.2 ± 0.7b ND ND 23.5 ± 2.5b 5.3 ± 1.5b

San Martín Pulp 58.1 ± 1.8c 23.2 ± 0.8b 13. 4 ± 0.5b 53. 8 ± 0.8c 7.9 ± 0.6ab
Villanueva Pulp 51.1 ± 0.4d ND ND 22.7 ± 1.6b 5.9 ± 1.1ab

San Martín Rind 70.3 ±0.1a 2.0 ± 0.1c 2.1 ± 0.1c 10.8 ± 0.9d 16.2 ± 3.1c
Villanueva Rind 126.0 ± 1.4d 0.4 ± 0.1d 0.7 ± 0.1d 9.9 ± 0.4d 39.3 ± 2.0d
Different letters in the same row indicate significant differences (P< 0.05). All results were expressed at wet weight.
ND: not detected

CONCLUSION
The juice of prickly pear San Martín showed higher concentrations of TP, betalains, AA and AOX,
however, the rind of prickly pear Villanueva has levels of TP 2 times superior and of AOX 4 times
higher than juice and pulp of San Martín. These results suggest that functional compounds present
in the rind of the fruit must be extracted and added to the juice, in order to elaborate a product as a
beverage that can offer better health benefits to the consumer. The antioxidant activity only
followed the same trend of the content of TP, but it did not seem to be related to the concentration
of betalains and AA present in the samples, because of that, it is possible that these compounds are
the main contributors of the AOX.

REFERENCES
[1]SAGARPA. Plan Rector. Sistema producto nacional nopal 2004.
http://www.sagarpa.gob.mx/agricultura/Publicaciones/SistemaProducto/Lists/NopalTuna/Attachments/1/pr
n_nopal.pdf. [5 October 2010].
[2] Stintzing FC. Herbach KM. Mosshammer MR. Carle R. Yi W. Sellappan S. Akoh CC. Bunch R. & Felker P.
2005. Color, betalain pattern, and antioxidant properties of cactus pear (Opuntia spp.) clones. J Agric Food
Chem 53: 442-451.
[3] Cassano A. Conidi C. Timpone R. Avella M. & Drioli E. 2007. A membrane-based process for the
clarification and the concentration of the cactus pear juice. J Food Eng 80: 914-921.
[4] Gillespie KM. & Ainsworth EA. 2007. Measurement of reduced, oxidized and total ascorbate content in
plants. Nat Protoc 2: 871-874
[5] Huang D. Ou B. Hampsch-Woodill M. Flanagan JA. & Prior RL. 2002. High-throughput assay of oxygen
radical absorbance capacity (ORAC) using a multichannel liquid handling system coupled with a
microplate fluorescence reader in 96-well format. J Agric Food Chem 50: 4437-4444.

2046
 A novel emulsifier from spinach with appetite regulation abilities

Marilyn Raynera, Sinan Cem Emekb, Karolina Gustafssona,c, Charlotte Erlanson-Albertsson,

Per-Åke Albertssonb

a
Lund University, Department of Food Technology, Engineering and Nutrition, Lund Sweden.
(marilyn.rayner@food.lth.se)
b
Lund University, Dept. of Biochemistry and Structural Biology, Centre for Molecular Protein Science.
c
Lund University, Appetite Control Unit, Dept of Experimental Medical Science, BMC, Lund Sweden

INTRODUCTION
Seeing that high-fat diets are obesity promoting, a strategy that strengthens the control of
appetite with dietary fat could be a means to counter act over consumption. Chloroplast
thylakoid membranes isolated from spinach have been found to inhibit pancreatic
lipase/colipase activity [1] and, when included in food, induce satiety signals. This effect is due
to their ability to reduce the rate of lipolysis through the inhibition of the lipase-colipase
complex. They have a strong affinity to oil which both prevents the lipolytic enzymes from
coming in close contact with its substrate and at the same time protects the thylakoids form
proteolytic enzymes present in gastric juices form digesting the thylakoids as quickly [2]. This
affinity also imparts thylakoids interesting emulsification properties. The objective of this
study was to characterise thylakoids’ ability to stabilise oil-in-water emulsions, and to study
their interfacial properties in light of their capacity to inhibit pancreatic lipase-co lipase activity
in vitro. As lipolysis is an inherently interfacial process it is important to quantify their
interfacial properties and to maximize the oil-water interfacial area covered by the thylakoids
in future food formulations.

MATERIALS & METHODS

Thylakoids isolated from spinach were used for emulsification studies using a lab-scale high
shear homogenizer. 33.3% vv oil-in-water emulsions produced had varying amounts of
thylakoids and resulting emulsions were characterized by creaming phase volume, microscopy
and light scattering to determine micro structure and droplet size distributions and surface load.

RESULTS & DISCUSSION

Electron micrographs (figure 1) show that thylakoids are attached to the surface of the oil
droplets in the form of intact thylakoids or sub-thylakoid membrane vesicles in the size range
1–2Pm. The resulting Emulsions were stable against coalescence but were subject to creaming.
There was very little difference between the emulsions at day 1 and after 7 days other than the
serum phase was slightly clearer after storage. The emulsification index was estimated by
image analysis and there was no significant change in the EI after 7days. EI and volume mean
droplet diameter as a function of thylakoids concentration is presented in figure 2. Drop size
distributions (d43) determined by light scattering showed very similar sized before and after
storage for all thylakoid concentrations with the exception of 2mg/ml oil, where the emulsions
after 7 days had a significantly smaller mean droplet size. In general, the stored emulsions had
a smaller droplet size which is opposite to what is generally expected. Perhaps there is some re-

11th International Congress on Engineering and Food - Athens, Greece, 2011 2047
organisation of the thylakoids at the oil-water interface and they become a more compact layer,
decreasing the measured size.
Thylakoid membrane segments /vesicles
are significantly larger than other
common emulsifiers, for example
surfactant molecules are in the range of
0.4 to 1 nm, and protein molecules are
between 1 and 5 nm [3]. Thylakoids are
least two orders of magnitude larger
than protein molecules, so it is not
unreasonable that we get surface
coverage in the range of 6 to 10 mg/m2
over the concentration range used. Figure 1: Micrograph of thylakoid stabilized emulsions

Figure 2: Volume mean diameter d43, mm (•) and
surface coverage, Emulsification index (¸), as a
function of thylakoid concentrations measured after 7
days storage at 4qC. Data points labelled with different
letters are significantly different(p<0.05).
CONCLUSION
Thylakoid membranes effectively
stabilize oil-in-water emulsions
which should facilitate their
incorporation in fat containing food
with satiety promoting effects. They
remained unchanged with respect to
cream layer height (emulsification
index), and droplet size distribution
over a storage period of 7 days,
indicating that despite their large
size, thylakoid stabilized emulsions
were quite stable. In future work it
would be of interest to increase to
even higher thylakoid concentration,
to the point that there is an excess. In
this way we could know the
maximum practical amount for
formulating foods with hunger
suppressing properties.

REFERENCES
[1] Albertsson PÅ, Köhnke R, Emek S C, Mei J, Rehfeld J F, Åkerlund H-E, Erlanson-Albertsson C,
Chloroplast membranes retard fat digestion and induce satiety: effect of biological membranes on
pancreatic lipase/co-lipase. Biochem J 401: 727-733 (2007).
[2] Emek, S.C., Åkerlund, H.-E., Clausén, M., Ohlsson,L., Weström, B.,Erlanson.Albertsson, C.,
Albertsson, P-Å. Pigments protect the light harvesting proteins of chloroplast thylakoid membranes
against digestion by gastro-intstinal proteases. Food hydrocolloids (2010),
doi:10.1016/j.foodhyd.2010.12.004
[3] Tcholakova S., Denkov N.D. & Lips A. 2008. Comparison of solid particles, globular proteins and
surfactants as emulsifiers. PCCP. 10(12), 1608-27.

2048
 Physico-chemical analysis, antioxidant capacity and vitamins of six ecotypes of Chilean
Quinoa (Chenopodium quinoa Willd.)

Margarita Mirandaa, Antonio Vega-Gálveza,b, Elsa Uribea, Jessica Lópeza, Enrique Martínezb, María José
Rodrígueza, Issis Quispea, Karina Di Scalac,d

Department of Food Engineering, Universidad de La Serena, La Serena, Chile (margmir@gmail.com)

Center for Advanced Studies in Arid Zones, CEAZA, Universidad de La Serena, La Serena, Chile
(avegag@userena.cl; enrique.a.martinez@ceaza.cl)
Food Engineering Research Group, Universidad Nacional de Mar del Plata, Mar del Plata, Argentina
(kdiscala@fi.mdp.edu.ar)
CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas). Argentina

INTRODUCTION
Quinoa (Chenopodium quinoa Willd.) is a native plant of the Andean region [1]. Quinoa has
gained an increasing interest in recent years due to its nutritional value as well as its
antioxidant capacity and phytochemical content [1,2]. The objective of this work was to
determine the physico-chemical properties, vitamins (B1, B2, B3 and E) and the antioxidant
activity of six ecotypes of quinoa (Chenopodium quinoa Willd) cultivated in three quinoa
production zones in Chile (North Highlands, Center and Southern Chile).

MATERIALS & METHODS

The quinoa seeds were harvested from the three ancestral production areas of Chile including
samples from the three genetic pools (North Highlands, Central and Southern Chile). The
samples are called Ancovinto and Cancosa from the North, Cáhuil and Faro from the Center,
and Regalona and Villarica from the South. Proximate analysis followed the recommendations
of the Association of Official Analytical Chemists [3]. Total phenolic content (TPC) was
determined colorimetrically using the Folin-Ciocalteau reagent (FC) according to Miranda et
al. [1] with modifications. Free radical scavenging activity of the samples was determined
using the 2,2,-diphenyl-2-picryl-hydrazyl (DPPH) method with some modifications [1].
Vitamin E was determined using HPLC with fluorescence detector, following the methodology
presented in Miranda et al. [1]. Vitamin B1 and B2 were extracted by acid hydrolysis and
enzymatic, determined Fluorometric method by HPLC, following A.O.A.C 942.23 and 970.65
[4] respectively. B3 was determined by spectrophotometry, according to A.O.A.C. 961.14 [4].
The Statgraphics Plus® 5.1 software (Statistical Graphics Corp., Herndon, USA) was used to
perform one-way analysis of variance (ANOVA) in order to determine significant differences
among samples (D=0.05).

RESULTS &DISCUSSION
Physico-chemical composition, total phenolic content (TPC), DPPH free radical scavenging
activity (IC50), vitamin B1, B2, B3 and vitamin E of six quinoa ecotypes from the three genetic
zones (North, Centre and South) are shown in Table 1. Proximate analysis of quinoa seeds
including moisture, crude protein, fat, crude fiber, ash and available carbohydrates for the six
ecotypes showed differences among samples (p-value<0.05). All cultivars showed good quality
with respect to protein; Regalona and Villarica from the South presented the highest content
(14.66 and 16.10 g/100 g, respectively). According to DPPH and TPC analysis, Faro and

11th International Congress on Engineering and Food - Athens, Greece, 2011 2049
Cáhuil from the Center showed the highest antioxidant capacity (p-value<0.05). Regalona
(South) presented the highest content in vitamins B1 (0.648 ± 0.006 mg/100 g) and B3 (1.569
± 0.026 mg/100 g) while the highest value of vitamin B2 (0.081 ± 0.002 mg/100 g) was
observed in Ancovinto samples from the North. Regarding vitamin E, the highest value was
shown by the Southern Villarica samples (4.644 ± 0.240 mg/100 g d.m.). The nutritional
composition and antioxidant activity values in the present study is comparable to the values
found in the literature for quinoa seeds, Miranda et al. [1], Vega-Gálvez et al. [2], Koziol [5].

CONCLUSION
The quinoa ecotypes have influence on the physico-chemical characteristics, antioxidant
activity, vitamins and therefore on their nutritive value (p-value<0.05). Regalona and Villarica
from the South presented the highest protein content (14.66 and 16.10 g/100 g, respectively).
According to DPPH and TPC analysis, Faro and Cáhuil from the Center showed the highest
antioxidant capacity (p-value<0.05). Southern Regalona presented the highest content in
vitamins B1 (0.648 ± 0.006 mg/100 g) and B3 (1.569 ± 0.026 mg/100 g) while the highest
value of vitamin B2 (0.081 ± 0.002 mg/100 g) was observed in Ancovinto samples from the
North. Regarding vitamin E, the highest value was shown by the Southern Villarica samples
(4.644 ± 0.240 mg/100 g d.m.).

REFERENCES
[1] Miranda M., Vega-Gálvez A., López J., Parada G., Sanders M., Aranda M., Uribe E. & Di Scala K. 2010. Impact of
air-drying temperature on nutritional properties, total phenolic content and antioxidant capacity of quinoa seeds
(Chenopodium quinoa Willd.). Industrial Crops and Products, 32, 258–263.
[2] Vega-Gálvez A., Miranda M., Vergara J., Uribe E., Puente L. & Martínez E.A. 2010. Nutrition facts and functional
potencial of quinoa (Chenopodium quinoa Willd.). An ancient Andean grain: A review. Journal of Science Food
Agricultural, 90(15), 2541-2547.
[3] AOAC, 1990. Official Method of Analysis. (15th Ed.), Association of Official Analytical Chemists. Washington,
D.C., USA.
[4] AOAC, 1995. Official Method of Analysis. (16th Ed.), Association of Official Analytical Chemists. Washington,
D.C., USA.
[5] Kozioá M.J. 1992. Chemical Composition and Nutritional Evaluation of Quinoa (Chenopodium quinoa Willd.).
Journal Food Composition and analysis, 5, 35-68.

2050
 Optimization Xylitol Production Conditions From Sunflower Stalk

Ozlem Akpinar, Reyhan Selin Uysal, Serdal SabancÕ, Burcu Sapci

Gaziosmanpasa University, Tokat, Turkey (oakpinar@gop.edu.tr)

INTRODUCTION
Xylitol is a five carbon sugar alcohol, equivalent to sucrose in sweetnes and occurs widely in
nature but it is also produced in human metabolism. Unlike sucrose, it is anticariogenic, natural
sweetener and can be consumed by diabetics because it is metabolized by an insulin-
independent pathway. It gives a pleasant cool and fresh sensation due to its high negative heat
of solution. [1]. Xylitol is used in various food products such as chewing gum, candy, soft
drinks and ice cream. Despite many advantages of xylitol, the use of xylitol as sweetener is
limited. Commercially, xylitol is produced from birch wood tree which is the most expensive
source. Agricultural wastes, widely available in Turkey are produced at an annual rate of more
than 50 million tons. These wastes can be used as animal feed, but this use has slight
economical significance. They are usually left to rot or burned in the field after harvesting [2].
Nowadays, utilization of these lignocellulosic wastes for industrial purposes receive enormous
attention due to their huge amount of carbohydrates (cellulose and hemicellulose) contents, low
cost, wide availability and reduction of environmental pollution. The agricultural waste which
is rich in lignocellulosic materials is an ideal source for the production of xylitol. Sunflower
stalk is one of the most widely available waste in Turkey. The biotechnical production of
xylitol employing hemicellulose fraction of lignocellulosic materials, instead of pure xylose is
more versatile approach to reduce the cost of production [3]. The aim of this study was to
produce xylose from sunflower stalk and optimization of production conditions of xylitol form
xylose by Candida tropicalis. The present study determined the effect of aeration, inoculum
and xylose concentration on the yield of xylitol from sunflower stalk and tobacco stalk.
Response surface methodology was used as a statistical design to optimize the formation of
xylitol in the hydrolysate.

MATERIALS & METHODS

Xylose Production: Hydrolysis of sunflower stalk was performed in a 1 L stainless-steel
pressure batch reactor. The reaction was carried in the range of 120 oC under 4% sulfuric acid
concentration and 30 minutes of residence times. Hydrolysate was analysed by HPLC on
Aminex HPX 87H (300 x 7.8 mm) column.
Xylitol Production: The fermentations were performed in a 1.5-L fermentor with agitation,
aeration, temperature, pH, and dissolved oxygen control. Experiments were carried out at 30°C
with 0.5 L of fermentation medium and 300 rpm. Samples were analysed by HPLC on
Aminex HPX 87H (300 x 7.8 mm) column. For optimization study the effects of air, substrate
and cell concentration on the production of yield and xylitol production rate were investigated.
Response surface methodology (RSM) was used for the optimization of xylitol production
conditions.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2051
RESULTS & DISCUSSION
Composition of sunflower stalk acid hydrolysate is presented in Table 1. The xylose
concentration in sunflower stalk hydrolysate, achieved at 120 oC for 30 min with 4% of acid
was 9.82 g/l.

Table 1. Composition of sunflower stalk acid hydrolysate

Component Amount
Xylose (g/l) 9.82+1.06
Glucose (g/l) 0.63+0.12
Arabinose (g/l) 0.14+0.18
Furfural (g/l) 0.22+0.08
Xylose yield (g xylose/ 100g max xylose) 36
Selectivity (g xylose/g glucose) 16
The optimization was conducted with the help of ‘Design-expert’ program. Optimization
method consists of overlaying the contour plots of both models. To optimal working conditions
based on high level of xylitol yield and volumetric xylitol production rate were chosen using
the following criteria: xylitol yield > 40 g/g (g xylitol produced/g xylose consumed) and rate >
0.15 g/L sa (g/l xylitol/time). The overlaying plot (Figure 1) shows the regions that shaded area
do not fit the optimization criteria while non-shaded area meet the optimization criteria.
Cell concentration (g/l)

Xylose concentration (g/l)

Figure 1. Overlaying plots of xylitol yield and volumetric xylitol production rate

CONCLUSION
The results showed that utilization of this material for production of xylitol does not only
solves the proper disposal of these wastes, but also provides additional income for farmers and
generates employment.

REFERENCES
[1] Winkelhausen, E. and Kuzmanova, S., 1998. Microbial conversion of d-xylose to xylitol. Journal of
Fermentation and Bioengineering, 86, 1-14.
[2] Bascetincelik, A., Ozturk, H.H., Karaca, C., Kacira, M., Ekinci, K., Kaya, D., Banan, A., Gunes, K.,
Komitti, N., Barnes, I. and Nieminen, M., 2006. Guide on Exploitation of Agircultural Residues in
Turkey Life 03 TCY/TR/000061.
[3] Parajo, J.C., Domingues, H. and Dominguez, J.M., 1998. Biotechnological production of xylitol. Part
1: interest of xylitol and Fundamentals of its biosynthesis. Bioresource Technology, 65, 191-20

2052
 Effect of fat substitution on the textural properties of cake

Vassiliki Psimouli, Vassiliki Oreopoulou

National Technical University of Athens, Athens, Greece
(psimouli@chemeng.ntua.gr, vasor@chemeng.ntua.gr)

INTRODUCTION
Ingredients which can substitute fat can be a helpful tool in weight control strategies.
Nevertheless fat substitution is a rather complicated matter since it affects the structure and the
organoleptic characteristics of the foodstuff. In the present research the effect of different
levels of fat substitution as well as the effect of different types of fat substitutes on the textural
behaviour of cake during compression was studied.

MATERIALS & METHODS

Four types of fat mimetic were used: maltodextrin of low dextrose equivalent, CdryLight
(Cargill); inulin, HP (Orafti); microparticulated whey protein, Simplesse and pectin, Slendid
(CPkelco). The fat mimetics were dissolved in cold water at concentration of 20% wt/wt,
except for Slendid and Simplesse which were dissolved at concentrations of 10% and 35%
respectively. The solutions were stored at 4°C overnight, so as to obtain gel formulations. All
fat substitute preparations were used to replace 35%, 65% and 100% of fat in cakes.
The textural behaviour was determined by the texture analyzer (TA-Xti2 Stable Microsystems,
Surrey, UK). Cake samples of 40x40x20mm size were compressed (test speed 1mm/s,
penetration distance 8mm) using the Sris P/75 aluminum plated probe. The measurements were
performed in triplicate, 20 hours after the cake preparation and the corresponding strain stress
data were reported. The sensory analysis was conducted by a ten member panel. Cake samples
were evaluated for total acceptance, crumb firmness, crumbliness and mouthfeel of
decomposition.

RESULTS & DISCUSSION

The strain stress data corresponded to a sigmoid pattern which was fitted by a mathematical
model reported by Swyngedau & Peleg [1], slightly modified.
V C 1 >1  exp  C 2 ˜ H @1 / C 3

İ ǻl
l0
Where ı is the stress, İ is the strain, which are derived from the force deformation data, and
constants C1, C2, and C3 are related to the shape of the sigmoid curve. The greater the value of
C1 the steeper the stress-strain curve, whereas C2 is a scale factor. The closer the value of C3 is
to 1 the existence of shoulder becomes less pronounced. All samples with fat substitutes
presented greater values of C1 compared to control, which is illustrated by a steeper stress-
strain curve and indicative of greater hardness and lower crumbliness of the texture.
Nevertheless the resistance to deformation of the fat substitute samples does not follow the
increase of fat substitution level, since in most cases the sample with 65% of fat mimetic
presented less compressibility than the respective samples of 100% substitution.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2053
 Simplesse
100% inHP
65%

Slendid
35% CdLight
control

Figure 1. Stress-strain curves of cakes containing a) inulin HP at different levels of fat replacement and
b) different fat replacers at 100% level of fat substitution.

This does not necessarily contradict the sensory evaluation which indicated that as the level of
substitution increases, the firmness increased, and the perceived crumbliness and mouthfeel
decomposition decreased. This discrepancy can be attributed to the significantly higher pore
size of samples containing 100% substitutes than the ones of 65%, resulting in a more fractural
structure.
A comparison of the effect of the different fat replacers on cake hardness, as evaluated by
stress-strain curves indicated that there was no significant difference at the level of 35% of fat
substitution whereas the greater differentiations were observed at 100% substitution (Figure
1b). Total fat replacement by Simplesse and Cdry Light lead respectively to the highest and
lowest resistance to deformation, which was also verified by the sensory analysis. The greater
tenderness provided by maltodextrin as fat substitute compared to other fat substitutes was also
observed in cookies formulation [2]. However, cake formulations of total fat substitution were
not evaluated as acceptable regardless of the fat mimetic.

CONCLUSION
The fat substitution affected the compressibility of the cake structure, leading to decreased cake
tenderness. Cakes of 35% of fat substitution demonstrated similar compressive behaviour to
control whereas the cakes of total fat replacement for some types of fat substitutes appeared to
be less resistant to deformation compared to the respective 65% ones, which can be attributed
to the presence of larger pores in its structure. The protein based fat substitute presented the
greater differentiation among the other fat mimetics in terms of its textural behaviour, by
presenting the higher resistance to deformation at total fat substitution.

REFERENCES
[1] Swyngedau S. & Peleg M. 1992. Characterization and prediction of the compressive stress-strain
relationship of layered arrays of spongy baked goods. Cereal Chemistry, 69(2), 217-221.
[2] Zoulias E.I., Oreopoulou V. & Tzia C. 2002. Textural properties of low-fat cookies containing
carbohydrate- or protein- based fat replacers. Journal of Food Engineering, 55, 337-342.

2054
 Determination of fructooligosaccharides (FOS) with FT-IR in cereals.

Their impact as substitute sweeteners in starch based desserts.

S. V. Protonotarioua, C. Pappasb, P.A. Tarantilisb, M. Polissioub, S. Yanniotisa, V. Evageliouc, I. Mandalaa

a
Laboratory of Engineering, Processing and Preservation of Foods, Department of Food Science and
Technology, Agricultural University of Athens, Greece (imandala@aua.gr)
b
Laboratory of Chemistry, Department of Science, Agricultural University of Athens, Greece
(ptara@aua.gr)
c
Laboratory of Food Chemistry, Department of Food Science and Technology, Agricultural University of
Athens, Greece (evageliou@aua.gr)

INTRODUCTION
Fructooligosaccharides (FOS) are considered alternative sweeteners [1]. Cereals are rich in
phytochemicals and fibers [2], thus the presence of FOS in them could enhance further their
profile. Furthermore, increasing FOS amount in starch based cream caramel desserts is
desirable, given their high nutritional value. Howver, it is difficult to determine in an accurate
way different FOS in a food item. The main objectives of this study were: a) to determine the
FOS amount in different milling fractions of wheat flour and b) to partially replace sucrose
with FOS in starch based desserts

MATERIALS & METHODS

Powder-caramel custard mixture, without sugar and milk, supplied by the greek company
JOTIS S.A., commercial sucrose powder by the Hellenic Sugar Industry S.A and high
pasteurization milk supplied by FAGE were used in order to prepare the desserts. Actilight
FOS, 95 r 2%, donated by Beghin Meiji was used for sucrose substitution. Wheat milling
fractions were donated by LOULIS S.A. and KAPLANIDIS GROUP.
Measurements involved the characterization and comparison of milling fractions with respect
to their FOS content by using Diffuse Reflectance Infrared Fourier Transform spectroscopy
(DRIFTs). Spectra collected between 4000-500 cm-1 (mid infrared), were smoothed and base
line corrected. A library (Wheat_Bran) was created using the OMNIC software ver.7.3. Cream
Caramel was made by progressively replacing its sucrose content with FOS (10-50%
substitution). Small-deformation oscillatory measurements were performed using parallel plate
geometry. Samples were cooled from 85°C to 5°C, at a fixed rate of 2°C/min. G' (elastic
modulus), G" (storage modulus) and Ș* (complex dynamic viscosity) were measured during
cooling at a fixed frequency of 1 Hz. At the end of cooling was recorded a variation of G', G"
and Ș* with frequency (Ȧ). Sensory evaluation of these products was also performed by a team
of trained panelists. The Triangle test was used as testing method.

RESULTS & DISCUSSION

Among the bands found, the most important was the one at 1158 cm-1which is a typical band
ascribed to the stretching of hydroxyl groups, found in oligosaccharides [3]. The characteristic
anomeric region absorption bands for b-linkage (939 cm-1) and a-linkage (898 cm-1) can be

11th International Congress on Engineering and Food - Athens, Greece, 2011 2055
unique for each sugar. The volume (area) of these picks was measured. In all cases the areas
found for wheat flour were greater than the respective ones for bran. Moreover, the spectrum of
FOS was recorded and compared with those of the Wheat_Bran library. All flours match more
than the respective brans with FOS According to the results found, wheat flour had a greater
FOS amount than coarse bran. Overall, these results showed that differences in FOS content in
mill fractions might be confirmed in a fast and simple way using DRIFTs.
Small-deformation oscillatory measurements were made in order to investigate the effect of
partial replacement of sucrose by FOS on the formation and the quality of the final products. A
first observation was that for all samples, G’ was greater than G’’ at the loading temperature of
85°C. Thus, the samples, although fluids, already had a substantial gel-like character at high
temperature. Moreover, the samples that sucrose was substituted by 20 and 30% FOS started at
much higher G’ values than the remaining two samples, which, on the other hand, exhibited
greater strength at 5°C. During the cooling part of this procedure, both elastic and viscous
characters initially increased rapidly with decreasing temperature followed by a region of a
much slower evolution. Finally, the network became weaker as temperature started to increase.
However, the substitution of sucrose by FOS led to a decrease not only on the gel character of
the gel but also on their strength/ solid-like character.
Differences in sweetness were evaluated by eleven trained assessors using triangle tests. Null
hypothesis is that assessors cannot determine a difference between the samples, in a
significance level of 5%, with probability Po=1/3. The difference was not significant between
the samples with 100% sucrose and 70 %sucrose-30% FOS. Thus, a substitution to a smaller
amount has not been investigated. In a descriptive test obtained for samples with a substitution
of sucrose at 50%, assessors found differences in sweetness in comparison to control samples.

CONCLUSION
Different milling fractions had different composition of FOS. Among the bands found, the
most important were the one at 1158 cm-1 and those of b and a anomeric. It indicated that wheat
flour had more FOS amount than bran.For all samples at the temperature of 85°C, G’ was
greater than G’’ showing that the sample, although fluid, already had a substantial gel-like
character. The recorded mechanical spectra showed that all samples shared a similar gel-like
character. In fresh samples panelists did not find significant differences between the samples
with 100% sucrose and 70 %sucrose-30% FOS. A further substitution at 50% resulted in
distinguish differences to control samples. A substitution of sucrose with FOS at 10% indicated
a sensorial quality stability in a short storage period for the cream caramel samples. Thus,
samples of similar sensory quality to the control ones can be produced at low FOS
replacement, which in addition have improved storage stability.

REFERENCES
[1] Yun, J.W. (1996). Fructooligosaccharides—occurrence, preparation, and application. Enzyme Microb.
Technol. 19, 107–117.
[2] Ranhotra G.S., Gelroth J.A and Astroth K. (1990). Total and Soluble Fiber in Selected bakery and
Other Cereal Products, Cereal Chem., Vol. 67, No. 5
[3] Kacurakova, M., Capek, P., Sasinkova, V., Wellner, N., & Ebringerova, A. (2000). FTIR study of
plant cell wall model compounds: Pectic polysaccharides and hemicelluoses. Carbohydrate Polymers, 43,
195–203.

2056
 The antioxidant properties of honey beer

Ana Kaluševiü, Gordana Uzelac, Mile Veljoviü, Saša Despotoviü, Mirjana Milutinoviü,
Ida Leskošek-ýukaloviü, Viktor Nedoviü

Department of Food Technology and Biochemistry, Faculty of Agriculture, University of Belgrade,

Nemanjina 6, 11080 Belgrade-Zemun, Serbia
(anakalusevic@gmail.com)

INTRODUCTION
Beer is a worldwide traditional natural drink which has a higher nutritional value than other
alcoholic beverages. It contains minerals and vitamins, proteins, organic acids and antioxidant
compounds, such as polyphenols. Among these antioxidants, phenolic compounds are of
particular interest to brewers because they play a key role in the brewing process by delaying,
retarding or preventing oxidation processes [1].
Honey is a natural food product well known for its high nutritional value. It has a wide range of
different constituents, including polyphenols, with significantly antioxidant properties. The
awareness of the therapeutic potential of honey is gradually growing and scientific evidences
of its effectiveness in several experimental and clinical conditions are beginning to emerge.
The objective of this study was to examine and compare phenolic profiles and antioxidant
activities of two different types of honey beers.

MATERIALS & METHODS

The wort and bottom-fermenting yeast used in this study were obtained from a local
brewery. Two types of honey, sunflower and linden honey, were purchased from local
market. Control beer was produced by fermenting pure wort without adding of honey.
Gallic acid, Folin-Ciocalteu’s phenol reagent, ammonium hydroxide, hydrochloric acid,
sodium acetate trihydrate, glacial acetic acid, ammonium ferric citrate and sodium carbonate,
carboxylmethylcellulose (CMC), sodium ethylendiamintetraacetate (EDTA) were purchased
from Merck (Germany). Ascorbic acid, 2,4,6-trypyridyl-s-triazine (TPTZ), ferric chloride
hexahydrate, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-
carboxylic acid, sodium dihydrogen phosphate, sodium hydrogen phosphate, sodium chloride
and potassium persulfate were purchased from Sigma-Aldrich (Germany).

Folin-Ciocalteau method: For the determination of total polyphenols the adjusted method
with Folin-Ciocalteau reagent was used [2]. After 2 h of reaction at room temperature, the
absorbance at 760 nm was determined.
EBC method: Absorbance of samples was measured on the spectrophotometer (Jenway 6400)
at wavelength Ȝ = 600 nm [3].
DPPH· method: Antiradical activity was measured after the reaction with free stable radical
1,1-diphenyl-2-picrylhydrazyl (DPPH࣭) according to Brand-Williams [4]. Absorbance is
measured at wavelength Ȝ = 515 nm on the spectrophotometer (Jenway 6400).
FRAP assay: Absorbance readings were made at 593 nm every 4 min. Results were expressed
as mM Fe(II)SO4 x 7H2O [5].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2057
2058
 Evaluation of green coconut (Cocos Nucifera L.) pulp for use as milk, fat and emulsifier

replacer in ice cream

Inês Aparecida Santana, Eliana Paula Ribeiro, Antonia Miwa Iguti

Maua Institute of Technology, São Caetano do Sul, Brazil (iasantana@maua.br)

INTRODUCTION
In Brazil, the 4th largest world coconut producer (around 2 billion fruits in 2010), green
coconut water (6-8 months) is widely consumed and the consumption reaches around 350
million liters per year in fresh and industrialized form. Despite the benefits of coconut water,
the coconut shell and the edible solid endosperm or pulp are discarded generating a huge
amount of waste. In our laboratory we have investigated this pulp as food ingredient. In
chocolate ice cream, e.g., it was efficient to replaced fat, milk, emulsifier and stabilizer. The
product was sensory approved by 93% of panelists, who judged its texture and organoleptic
properties very similar to true ice cream. More research is necessary regarding the composition
and properties of this material, to understand its air incorporation capacity in ice cream. The
objective of this study was to characterize and evaluate functional properties of fresh and
freeze-dried pulp from green dwarf coconut (Cocos nucifera L.), using umbu ice cream system
to evaluate the influence of lipids on product quality. Umbu (Spondias tuberosa) is a
bittersweet fruit of Brazilian Northeast, was chosen to verify if, at low pH values, the coconut
pulp could give the same good results as those obtained with chocolate

MATERIALS & METHODS

Green coconuts (Cocos nucifera L.) were from Brazil Northeast. Fresh pulp (FP) and freeze-
dried pulp (FD) proximate composition was performed according to AOAC standards. Lipid
content in FP and FD was determined by Bligh Dyer and percolation, respectively. The fatty
acid composition was determined by GC after methylation of the oil extracted from FD.
Foaming capacity was determined for FP, FD and defatted freeze-dried pulp (DFD) by
measuring the volume of foam formed in the presence of baking powder and water at 40 ºC
and compared with egg white. Emulsifying capacity of FP, FD, DFD was determined by
measuring the volume of oil required to cause the emulsion inversion in water, and compared
with egg yolk. Two formulations of ice cream were prepared: one with 5.5% FD, 20% umbu,
20% sucrose, 54.5% water; in the other, FD was replaced by DFD. Hardness [1], meltdown [2]
and overrun were determined.

RESULTS & DISCUSSION

The proximate composition were 92.70% and 8.01% moisture, 0.39% and 27.95% fat, 0.97%
and 19.9% protein, 0.75% and 10.72% ash and 5.19% and 33.42% carbohydrate (by
difference), respectively for FP and FD. The main component of dry matter pulp is
carbohydrate. The low fat content is expected for green dwarf and is in accordance with the
studies of Aragão et al. [3].Table 2 gives fatty acidy composition, that is typical for coconut oil,
with predominance of medium chain fatty acids. The fatty acid profile is similar to results of
Aragão et al. [3] for fruits with 6th to 8th maturity month. Foam expansion or overrun, resulted
in 120% for FP, 209% for egg white, 77.6% for FD and 70.8% for DFD. The FD pulp showed

11th International Congress on Engineering and Food - Athens, Greece, 2011 2059
lower foaming capacity but higher stability than FGP and egg white. The collapse of foams
produced by FP and egg white occurred in shorter time, probably because of greater
concentration of protein in the FD, resulting in smaller bubbles and higher viscosity [4].

Table 2. Fatty acid composition of coconut lipids

Fatty Caproic Caprylic Capric Lauric Myristic Palmitic Stearic Oleic Linoleic
acid (C6) (C8) (C10) (C12) (C14) (C16) (C18) (C18:1) (C18:2)
% 0.97 5.10 3.58 38.05 20.10 15.20 2.35 12.52 2.13

For emulsifying capacity, the volumes of oil to cause inversion of the emulsion were 72.7
mL/g (dry basis), 19.17 mL/g, 28.9 mL/g and 28.3 mL/g, respectively for FP, egg yolk, FD and
DFD. The lipids showed no influence in this property, which indicates that the responsible for
emulsifying capacity in this system are proteins, that can be influenced by many factors such as
type of protein, pH, ionic strength, temperature, protein concentration and solubility [4, 5]. Ice
cream. The product made with FD showed 28.36% of overrun and addition of DFD, 17.15% of
overrun. Low values for overrun resulted of lab scale ice cream maker used to process. The
results showed that without lipids a reduction of the overrun value (11%) occurred. When FP
was used in ice cream formulation, the overrun was higher than with FD and DFD and similar
to umbu ice cream milk-based (data no shown). The hardness of ice cream made with FD was
51% lower than that made with DFD, so there is a reverse relation between fat content and
hardness. Meltdown showed small differences. The half life time was 48.9 and 45.86 min for
FD and DFD, respectively.

CONCLUSION
Green coconut pulp has foaming and emulsifying capacities that can be used for production of
ice cream, even at low pH values. As FP is almost tasteless and odorless, it is appropriate to
formulate ice cream-like products. Medium chain fatty acids predominate in coconut oil,
mainly lauric acid. The lipids had more influence on foaming capacity, overrun and hardness
than in emulsifying capacity and meltdown. There are potential applications of green coconut
pulp as milk, fat and emulsifier replacer in ice cream and other products, e.g., bakery. It is a
healthy, sustainable and economical alternative to countries that are producers of this crop.

REFERENCES
[1] Soukoulis, C.; Chandrinos, I.; Tzia,C. 2008. Study of the Functionality of Selected Hydrocolloids and Their Blends
with ț-carragean on Storage Quality of Vanilla Ice Cream. Food Science and Technology, 41, 816-1827.
[2] Roland, A. M.; Phillips, L. G.; Boor, K. J. 1999. Effects of Fat Content on the Sensory Properties, Melting, Color
and Hardness of Ice Cream. Journal of Dairy Science, 82(1), 32-38.
[3] Aragão, W. M.; Cruz, E. M. de O; Tavares, M.; Ribeiro, F. E.; Tupinambá, E. de A.; Pimentel, S. A. &
Takemoto,E. 2004. Teor de Gordura e Composição de Ácidos Graxos em Polpa de Frutos de Coqueiro Anão em
Diferentes Idades de Maturação. Revista do Instituto Adolfo Lutz, 63(2), 159-167.
[4] Damodaran S. 2008. Amino Acids, Peptides and Proteins. In: Damodaran S.; Parkin K. L. & Fennema O.R. (Eds.).
Fennema’s Food Chemistry, 4th ed. CRC Press Taylor & Francis Group, Boca Raton, Florida, USA. p.1144.
[5] Kwon, K. S. & Rhee, K. C. 1996. Emulsifyng Capacity of Coconut Proteins as a Function of Salt, Phosphate, and
Temperature. JAOCS, 73(12) 1669-1673.

2060
 Antioxidant activity and phenolic content of extracts from different Pterospartum

tridentatum populations growing in Portugal

Maria Teresa Coelhoa, José Carlos Gonçalvesa, Vítor Alvesb, Margarida Moldão-Martinsb

a
Escola Superior Agrária de Castelo Branco, Quinta Sra de Mércules, Apartado 119,
6001-909 Castelo Branco (mteresacoelho@ipcb.pt)
b
CEER – Biosystems Engineering. ISA. Technical University of Lisbon. Tapada da Ajuda. 1349-017
Lisboa, Portugal (mmoldao@isa.utl.pt)

INTRODUCTION
Pterospartum tridentatum L. Willk. is an European endemic Leguminosae and known as
carqueja in Portugal. This small shrub is very common in the mountains of the north of
Portugal. Bioactive compounds, such as alkaloids and flavonoids, have been identified in
aqueous extracts of those plants [1]. Plants synthesize antioxidant compounds, as secondary
products, which are mainly phenolic compounds serving in plant defense mechanisms to
counteract reactive oxygen species (ROS) in order to avoid oxidative damage. Researchers are
looking for natural antioxidants as alternative to synthetic antioxidants.
Some authors refer the use of Pterospartum tridentatum in popular medicine and culinary uses.
This plant is an underexploited natural source of compounds with biological activity, which
should be fully characterized aiming to its valorization.
The aim of the present study was to evaluate the total phenolic content and antioxidant activity
of aqueous extracts of Pterospartum tridentatum samples, collected in three locations in
Portugal, at different vegetative stages.

MATERIALS & METHODS

Samples of the aerial parts of Pterospartum tridentatum were collected at different vegetative
stages: dormancy period (end of January) and flowering period (in May), in three locations in
Portugal: Orvalho, Gardunha mountain and Malcata mountain. Aqueous extractions were
performed by refluxing during 2 hours in a Clevenger apparatus. The extract solutions were
freeze-dried and a solid extract was recovered.
The antioxidant activity of the solid extracts was determined by the radical scavenging activity
method using 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). The total phenol content (TP) of
the extracts was evaluated by spectrophotometric method and expressed as gallic acid
equivalents (mg/g of dry mass). All trials were carried out in triplicate. The data were subjected
to one-way analysis of variance (ANOVA) and the differences between means were measured
using Duncan’s Test through STATISTICA, Version 7 (Copyright© StatSoft, Inc.), pvalues <
0.05 were considered to be significant.

RESULTS & DISCUSSION

The influence of the seasonal variation in the yield and composition of the extracts was
evaluated, in order to select the most appropriate harvest season. The extraction yields
presented some differences with the harvest period and the highest yield extraction was

11th International Congress on Engineering and Food - Athens, Greece, 2011 2061
obtained in the flowering period, using flowers (19.4g/100g plant dry mass in Gardunha
mountain) and the lowest extraction yield was also obtained in the same period but using
stems (11.3 g extract/100 g plant dry mass in Malcata mountain). The extraction yield was
always higher than the commonly used herbs [2].
The 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) is widely used to evaluate the antioxidant
capacity of extracts from different plant materials. The stem extracts showed significant
differences between the dormancy and the flowering period. At flowering stage a higher
antioxidant activity was observed in the flower extracts.
The total phenolic content of Pterospartum tridentatum (ranged from 270.7 to 402.9 mg gallic
acid equivalents per g dry matter) show very high levels at any time of harvest. The highest
value occurred in the dormancy period in Malcata mountain and are superior to other species
previously studied, like Harpephyllum caffrum in leaf and stem bark and Sclerocarya birrea in
stems [3]. They also contain much more phenolic compounds when comparable with results in
Carissa opaca [4] or in Australia herbs and spices like Tasmannia pepper leaf, anise myrtle and
lemon myrtle [5]. Plants with high levels of phenolic compounds have demonstrated a high
antioxidant activity of plants extract.
From preliminary experiments, it is anticipated a significant antimicrobial activity of the solid
extracts against bacteria and fungus (data not shown).

CONCLUSION
The studied Pterospartum tridentatum aqueous extracts present a high extraction yield, an
appreciable level of total phenolic compounds and a significant antioxidant activity. The results
foresee a high potential for the utilization of this plant or its extracts as a new source of safe
natural antioxidants and preservatives for the food industry with consequent health benefits for
consumers. From the results, it can be conclude that the plants can be harvested at all seasons
of the year, which presents an advantage from an industrial point of view.

REFERENCES
[1] Vítor, R., Mota-Filipe, H., Teixeira, G., Borges, C., Rodrigues, A., Teixeira, A. & Paulo, A. 2004.
Flavonoids of an extract of Pterospartum tridentatum showing endothelial protection against
oxidative injury. Journal of Ethnopharmacology, 93, 363-370.
[2] Luís, A. Domingues, F., Gil, C. & Duarte, A.P. 2009. Antioxidant activity of extracts of Portuguese
shrubs: Pterospartum tridentatum, Cytisus scoparius and Erica spp. Journal of Medicinal Plants
Research, Vol.3(11), 886-893.
[3] Moyo, M., Ndhlala, A.R., Finnie, J.F. & Staden, J.V. 2010. Phenolic composition, antioxidant and
acetylcholinesterase inhibitory activities of Sclerocarya birrea and Harpephyllum caffrum
(Anacardiaceae) extracts. Food Chemistry, 123, 69-76.
[4] Sahreen, S., Khan, M.R. & Khan, R.A.. 2010. Evaluation of antioxidant activities of various solvent
extracts of Carissa opaca fruits. Food Chemistry, 122, 1205–1211.
[5] Konczak, I., Zabaras, D., Dunstan, M. & Aguas, P. 2010. Antioxidant capacity and phenolic
compounds in commercially grown native Australian herbs and spices. Food Chemistry, 122, 260–
266.

2062
 Physicochemical Characterization of Monoacylglycerols from Sunflower Oil

Galúcio, C. S.a; Souza, R. A.a; Stahl, M. A.b; Sbaite, P.a; Benites, C. I.a; Wolf Maciel, M. R.a

a
Laboratory of Separation Process Development (LDPS), School of Chemical Engineering (FEQ)
b
Laboratory of Oils and Fats, School of Food Engineering (FEA)
University of Campinas (UNICAMP), Campinas/SP, Brazil (cley.cie@hotmail.com)

INTRODUCTION
The monoacylglycerols (MAG) are emulsifiers with food, pharmaceutical and cosmetic wide
application. In these industries, MAG representing about 70% of all synthetic emulsifiers used.
Looking for the replacement of the artificial for natural ingredients, new researches are aimed
to obtain natural emulsifiers from fat and oils with unsaturated fatty acids.
Sunflower oil is distinguished by having large amounts of unsaturated fatty acids, such as oleic
and linoleic acids (Ȧ-6). The latter is of great importance because it is considered an essential
fatty acid.
The MAG are obtained through interesterification or glycerolysis of triacylglycerol (TAG)
with glycerol (GL) excess. In this reaction, the mixture obtained showed around 40 – 50% of
MAG, besides diacylglycerols, TAG, free fatty acids and GL. The aim of this work is to
characterize sunflower oil and acylglycerols mixture produced by chemical glycerolysis,
evaluating the product composition and quality.

MATERIALS & METHODS

Chemical glycerolysis of sunflower oil was conducted in jacketed glass reactors (2 liters). The
conditions applied in these reactors were: 200°C of temperature, 1% of Ca(OH)2 as catalyst
(per reagents mass), GL/TAG molar ratio of 4 and 60 minutes of reaction time [1]. The High
Performance Size Exclusion Chromatography (HPSEC) was used for the Acylglycerols content
[2,3].
Others analyses were according AOCS Official Methods [4]: Fatty Acids Composition (Ce1-
62); Determination of Mass per Unit Volume (Cc10c-95); Iodine Value (Cd1c-85);
Saponification Value (Cd3a-94); Free Fatty Acids (Ca5a-40); Peroxide Value (Cd8b-90);
Moisture (Ca2e-84).
Differential Scanning Calorimetry (DSC): differential scanning calorimeter (823E, Mettler
Toledo). The samples were analyzed under a flow of nitrogen gas (50 mL/min). A dynamic
scan was performed at a heating rate of 10°C/min over a temperature range of -100 to 600°C.

RESULTS & DISCUSSION

The acylglycerols mixture showed the following composition: TAG (6.9%), DAG (42.3%) and
MAG (48.3%). Free fatty acids weren’t detected and GL corresponded to 2.5%. The content of
MAG in the mixture was between 40 and 50% according to the references [1].
In relation of fatty acid composition, the unsaturated fatty acids are majority in sunflower oil
and the content of the oleic and linoleic acids, before and after the glycerolysis, are
approximate (around 35% of oleic and 51% of linoleic acid), indicating that chemical
glycerolysis in used conditions were satisfactory.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2063
The results of physicochemical characterization of sunflower oil and glycerolysis product
(characteristics of them) are shown in the Table 1. According CODES standard [5], the results
obtained to refined sunflower oil were used as reference in this research, and they are within
the standard.

Table 1. Characteristics of sunflower oil and glycerolysis product

Analysis Sunflower oil Mixture of acylglycerols
Mass per Unit Volume (g/cm3) 0.8970 0.9449
Calculated Iodine Value (g I2/100g) 122.8 120.5
Calculated Saponification Value (mg KOH/g) 191.7 191.9
Free Fatty Acid (%) 0.262 1.061
Peroxide Value (mEq O2/kg) 0.669 1.530
Moisture (%) 0.026 0.345

Regarding the thermal analysis, the DSC curve profile (thermogram), showed for sunflower
oil, 2 endothermic peaks: the first one around -25ºC (melting process), and the second around
425ºC (vaporization process). For acylglycerols mixture, the thermogram shows also 2
endothermic peaks: the first one around -10ºC and the second around 425ºC like as oil.

CONCLUSION
The acylglycerols mixture from sunflower oil shows 48% of MAG, suitable for this type of
reaction. The results of fatty acid composition were similar to the sunflower oil and to the
product of chemical glycerolysis, noting that the major fatty acids (oleic and linoleic) were not
degraded in the reaction.
Following the accepted parameters of physicochemical and thermal properties for sunflower
oil, the results obtained for acylglycerols mixture showed acceptable standards. So, the
chemical glycerolysis is promising for natural MAG production and studies for further
concentration should be considered, seeking an industrial application of this product.

REFERENCES
[1] Galúcio, C. S.; Benites, C. I.; Sbaite, P.; Wolf Maciel, M. R. 2010. Monoacylglycerols obtaining from
sunflower oil. In: 19th International Congress of Chemical and Process Engineering CHISA 2010,
2010, Praga. CD-ROM of Full Texts, 2294-2298.
[2] Schoenfelder, W. 2003. Determination of monoglycerides, diglycerides, triglycerides and glycerol in
fats by means of gel permeation chromatography [C-VI 5b(02)]. Eur. J. Lipid Sci. Technol, 105, 45-
48.
[3] Arzamendi, G.; Arguiñarena, E.; Campo, I.; Gandía, L. M. 2006. Monitoring of biodiesel production:
Simultaneous analysis of the transesterification products using size-exclusion chromatography.
Chemical Engineering Journal, 122, 31-40.
[4] A.O.C.S. Official methods and recommended practices of the American Oil Chemist’s Society, 5th
ed. Champaign: AOCS, 1998.
[5] CODEX - Current Official Standards. CODEX standard for named vegetable oils. Available in:
<www.codexalimentarius.net/download/standards/336/CXS_210e.pdf> Accessed: February 17, 2010.

2064
 Antioxidant activity of the polyamines spermine and spermidine in soybean oil

Adriana Correa Mendonça & Maria Beatriz A. Gloria

LBqA – Laboratório de Bioquímica de Alimentos, Faculdade de Farmácia, UFMG, Av. Antônio Carlos,
6627, CEP 31270-901, Belo Horizonte, MG, Brasil (mbeatriz@ufmg.br)

INTRODUCTION
Lipid oxidation reactions are frequent in foods, and are often associated with food
deterioration. Even though it starts in the lipid fraction, eventually other food components are
affected, changing several desirable properties, among them, sensory, nutritional and functional
qualities. Furthermore, undesirable compounds can be formed and affect consumers’ health. It
also limits the shelf life of lipids and of foods containing significant amounts of lipids.
Soybean oil is widely used in Brazil because of its high availability and low price. Since it
contains high levels of unsaturated fatty acids, it is susceptible to oxidation. Therefore, it is an
interesting matrix to investigate ways to prevent lipid oxidation, and to determine which
antioxidants would be effective in preventing oxidation. Today, there is a tendency to
substitute synthetic antioxidants by natural ones. Among antioxidants which are natural in
biological systems, the polyamines spermine (EPM) and spermidine (EPD) play an important
role. They are soluble in aqueous and organic phases what facilitates the application in
different food matrices [1-3]. The objective of this study was to investigate the antioxidant
activity of spermine and spermidine individually and to compare the results with those of
traditional synthetic antioxidants and different combinations using Rancimat®.

MATERIALS & METHODS

Spermidine (EPD) and spermine (EPM) were purchased from Sigma Chemical Co. (St Louis,
MO, USA) and BHA and BHT were from Synth (Diadema, SP, Brazil). The antioxidant free
soybean oil was provided by Cargill Agrícola (Mairinque, SP, Brazil). The antioxidant activity
of the compounds in antioxidant free soybean oil was determined using a Rancimat® (model
743, Metrohm, Herisau, Swiss), according to the AOCS method Cd 12b-92 [4,5]. The standard
solutions were incorporated into the antioxidant free soybean oil using an ultrasound shaker
(UltraSonic Cleaner, Unique, SP, Brazil). The control was also kept in the shaker for the same
time period. The analyses were performed in duplicate. The experiments were performed at
110 ºC, at an air flow of 20 L/h, and 60 mL Milli-Q water was used for the collection of the
volatiles. Conductivity curves versus time and the induction periods were automatically
obtained. The results were expressed as protection factors (PF = ratio between the induction
periods of the sample containing the antioxidant and the control sample). The results were
analyzed with STATISTICA 8.0 (STATSOFT, USA), at 5% probability. The antioxidant
activity of the polyamines was compared with those of the synthetic antioxidants BHA and
BHT at concentrations of 0.01, 0.02, 0.03, 0.04, 0.05 and 0.06 g/100 g. The antioxidant
activity of different combination of EPM, BHA, and BHT at 1:1 (v/v) was determined.

RESULTS & DISCUSSION

The antioxidant activity of the amines EPD and EPM in soybean oil was confirmed. The
activity of EPM was higher compared to EPD. These results are similar to those reported by

11th International Congress on Engineering and Food - Athens, Greece, 2011 2065
Drolet et al. [2] and Løvaas [3], who concluded that the antioxidant activity of these
compounds was associated with the number of amine groups in the molecule. There was a
significant linear correlation (R2 = 98.16%) between the levels of the amines and the protection
factors. Based on this result, a response surface model was generated and can be used to
predict the shelf life of soybean oil at 110 °C, at concentrations from 0.0059 to 0.0341 g/100 g
oil using the equation: PF (protection factor) = 2.11 + 0.59EPM + 0.30EPD. Within the
concentration range investigated, there was no synergistic effect, but an additive effect of EPD
and EPM in soybean oil. The protection factor obtained for EPM was higher compared to
those obtained for BHA, BHT, BHA+BHT, EPM+BHA, and EPM+BHT (Figure 1).

Figure 1. Protection factors for soybean oil at 110 °C determined in Rancimat in the presence of
different concentrations of BHA, BHT, EPM and their mixtures at 1:1.

CONCLUSION
The polyamines EPM and EPD increased the oxidative stability of soybean oil. EPM was
more effective than EPD. The polyamines EPM and EPD showed higher antioxidant activity
compared to BHA and BHT. Therefore, the polyamines EPM and EPD are potent antioxidants
in soybean oil and provide higher protective factors than some of the traditional synthetic
antioxidants tested.

REFERENCES
[1] Ramalho V.C. & Jorge N. 2006. Antioxidantes Utilizados em Óleos, Gorduras e Alimentos
Gordurosos. Química Nova, 29, 755-760.
[2] Drolet G., Dumbroff E.B., Legge R.L. & Thompson J.E. (1986). Radical Scavenging Properties of
Polyamines. Phytochemistry, 25(2), 367-371,
[3] Løvaas E. (1997). Antioxidative and Metal Chelating Effects of Polyamines. Advances in
Pharmacology, 38, 119-149.
[4] Antolovich M., Prenzler P.D., Patsalides E., McDonald S. & Robards K. 2002. Methods for Testing
Antioxidant activity. Analyst, 127, 183-198.
[5] AOCS (2008). Method Cd 12b-92. Official Methods and Recommended Practices of the American
Oil Chemists’ Society, 5th edition. Champaign, IL: AOCS.

2066
 Encapsulation of natural flavors for use in dairy products

Santos, Sandra Diasa, Ressurreição, Sandrine Matiasa, Marques, Rui Ferreiraa,

Santos,Cristiana Valentea, Silva, Aida Moreiraa, Pintado, Manuela Estevezb

a
Escola Superior Agrária de Coimbra (ESAC), Instituto Politécnico de Coimbra, Portugal
b
Escola Superior de Biotecnologia (ESB), Universidade Católica Portuguesa, Portugal
Sds@esac.pt

INTRODUCTION
Over the years, the lifestyle has undergone considerable changes. From the standpoint of food,
it is known that the time spent preparing meals is markedly reduced. Allied to this situation, we
found consumers are increasingly demanding about the quality and diversity of products
looking to see associated flavor and properties (biological properties: antioxidant,
antimicrobial) that contribute to their health and quality of life.
Many flavor components are volatile which are susceptible to loss by evaporation, oxidation or
ingredient interactions. As a result, it is beneficial to encapsulate the volatile flavors prior to
use in foods. Encapsulation can be defined as any method employed to entrap a flavor in a
carrier to convert it to a more useful form or to impart some degree of protection against
evaporation, reaction or oxidation in food (Edris, A. and Bergnst‫ޣ‬hl, B., 2001). Of the different
types of encapsulation methods (coacervation, extrusion, spray-drying, multiple emulsions,
molecular inclusion, etc.), the use of cyclodextrins (method of molecular inclusion), has arouse
great interest in the scientific community, both in terms of research as the field of applied
technologies (ingredients of drugs in food or cosmetics). ȕ–Cyclodextrin (ȕ-CD) is a short,
hollow, truncated cone shaped molecule, which is formed by seven Į(1-4) linked gluco-
pyranoses in normal chair conformations (Silva et al, 2007) ȕ–CD is widely utilized in food
and pharmaceutical industries to encapsulate compounds that are sensitive to the environment,
have a slow solubility in water and high volatility.
Escola Superior Agrária de Coimbra (ESAC) - Instituto Politécnico de Coimbra, Portugal, has
a Dairy homemade. In this sense, the fresh aromatic plants and the essential oils encapsulated
were applied to some products produced there like fresh cheese, spread cheese and butter.
Sensorial analyses were made to avail the receptivity of the consumer to these products.

MATERIAL AND METHODS

The plants: parsley (Petroselium crispum var. latifolium), Coriander (Coriandrum sativum var.
microcarpum) and bush basil (Ocimum minimum) were obtained in ESAC fields.
Fresh leaves of coriander, parsley and bush basil were subjected to hidrodestyllation in a
Clevenger apparatus coupled a microwave in a minimum of twenty minutes. The essential oils
of parsley and coriander were obtained in a yield of 0.05% (w/w) and the essential oil of bush
basil was obtained in a yield of 1%.
The assessment of total antioxidant capacity was determined by ABTS•+ (Gião et al, 2007),
with modifications (evaluation of antioxidant activity in essential oil). This method is based on
the decolorization of the radical cation ABTS•+, measured as percentage inhibition after
spectrophotometer readings at 734 nm.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2067
In order to obtain an absorbance value (Abs) between 680 nm and 720 nm at a wavelength of
734 nm proceeded to the dilution of ABTS•+ solution in ultra-pure water. Each experimental
sample was added to 1 mL of ABTS•+, using such volume (10 mL), which after 6 minutes of
reaction the percentage of inhibition is between 20 and 80%. Measurement (oil dissolved in a
mixture 50:50 (ethanol: toluene) was performed in triplicate, with the final value was the
average of three replicas. The total antioxidant capacity was expressed as percentage inhibition
(PI), according to the equation PI = ((ABTS•+ Abs - Abs sample) / Abs ABTS•+) × 100 where
Abs ABTS•+ is the initial absorbance of diluted ABTS•+ and Abs sample the absorbance of
the sample after 6 minutes of reaction . The calibration curve was prepared in parallel using
standard solutions of ascorbic acid at various concentrations and under the same experimental
conditions. The final results are expressed as equivalent grams of ascorbic acid per liter of
sample (g/L).
The inclusion complex of cyclodextrins powders was prepared in a glass beaker. The ȕ-
Cyclodextrin (KLEPTOSE, was kindly offered by Roquette, France) was weighed and mixed
with distilled water. After dissolution, essential oil was added. The solution was mixed by
magnetic stirrer and stirred at 35 ºC for 1 h. The solution was cooled to refrigerate temperature
for 24 h. The precipitate was dry at room temperature.

RESULTS AND DISCUSSION

The value of total antioxidant activity of coriander essential oil equivalent of ascorbic acid
ranges from 0.025 ± 0.002 g/L, between 0.019 ± 0.002 g/L for the essential oil of parsley and
0,022 ± 0,002 g/L for the essential oil of bush basil.
Fresh cheese and spread cheese with essential oils encapsulated have been preferred in respect
products that were added fresh plants. Butter with encapsulated and with fresh plants was also
accepted.

CONCLUSIONS
Fresh cheese and spread cheese with essential oils encapsulated have been preferred in respect
products that were added fresh plants. Butter with encapsulated essentials oils and fresh plants
was also accepted. The extracts- essential oil with cyclodextrins prepared (“powder”) were
easily homogenized in the food product, but with fresh plants uniform size of plants and a
provision in homogeneous products has not been achieved.

REFERENCES
[1] Edris, A., Bergnst‫ޣ‬hl, B. (2001). Encapsulation of orange oil in a spray dried double emultion.
Nahrung/Food, 45, No. 2, pp. 133-137.
[2] Gião, Maria S.; Gonzaléz-Sanjosé, M. L.; Muñiz, Pilar; Rivero-Pérez, M. D.; Kosinska, Monika;
Pintado, Manuela E.; Malcata, F. Xavier (2007). Infusions of Portuguese medicinal plants:
Dependence of final antioxidant capacity and phenol content on extraction features. Journal of the
Science of Food and Agriculture, 87: 2638-2647.
[3] Silva, J., Galhano, C., Silva, A. (2007). A new sprout inhibitor of potato tuber based on carvone/ȕ-
cyclodextrin inclusion compound, J Incl Phenom Macrocycl Chem, 57, 1-4, 121-124.

2068
 Effects of dietary fiber on structure formation in bread during baking process

Annalisa Romanoa, Elena Torrieria,b, Paolo Masia,b, Silvana Cavellaa,b

a
CAISIAL-Centre of Food Innovation and Development in the Food Industry -University of Naples
Federico II – Naples, Italy (annalisa.romano@unina.it)
b
Department of Food Science -University of Naples Federico II –Naples, Italy

INTRODUCTION
The link between the intake of dietary fiber (DF) and health benefits has prompted the interest
in fiber-enriched foods such as fiber-enriched baked goods. A daily intake of approximately
30g is encouraged to promote health benefits associated with fiber. However, fiber intake is
commonly lower than the recommended one, as consequence the development of foods with
high fiber content should be desirable.
The aim of this work was to evaluate the potential use of two DFs (coffee silverskin and inulin)
at elevated content (9.8 %) as ingredient in breadmaking. The effects of DFs on dough
rheological behaviour, leavening and baking performance were studied by means of
rheological characterization of the dough behaviour and Image Analysis.

MATERIALS & METHODS

All samples were prepared in a Brabender farinograph by mixing wheat flour, water, salt (1.25
g), sugar (0.5 g) and yeast (0.75 g). The DFs used were CS- coffee silverskin and LI- long
chain inulin (with DP minimum 23) at 9.8% concentration (w/w).
Doughs were submitted to a lubricated squeezing test by means of an Instron Universal Testing
Machine. The deformation force was recorded as a function of time. Stress-strain curves for
biaxial extensional flow were derived according to [1]. Dynamic mechanic measurements were
carried out by means of a strain controlled rheometer, equipped with parallel-plate geometry.
All measurements were performed in oscillatory mode. Temperature ramp tests were
performed between 10 and 90°C, at a heating rate of 1°C/min. During tests the storage modulus
(G') as a function of temperature was monitored. Dough evolution during leavening and baking
phases was studied by measuring the variation in time of the total volume of the sample by
means of image analysis. Statistical analysis (ANOVA and Duncan test) were performed on
volume expansion ratio (volume at time t / initial volume) of samples.

RESULTS & DISCUSSION

Flour replacement at elevated levels by DFs changes rheological behaviour of dough and
breadmaking performances. Leavening and baking processes are characterized by fast biaxial
expansion of gas cells. The effect of DFs on biaxial extensional properties of wheat flour
doughs was investigated. For all cases detected, the curves can be divided into three sections:
pre-yield, yield and strain hardening. The CS and LI doughs have a higher yield stress and a
higher resistance to flow compared to control. The results reported illustrate that the squeezing
results are greatly depends on fibre source: CS shows the maximum strain hardening. During
baking, the dough undergoes severe irreversible changes that greatly affect its rheological
behaviour. The results prove that when DFs are added to the dough, they interact with water,
reducing the liquid water for starch. When CS is added, this phenomenon is related to a

11th International Congress on Engineering and Food - Athens, Greece, 2011 2069
decrease in the water ability due to increased fiber -water interaction. While the behaviour of
doughs with inulin may be attributed to gel-forming property and capability of inulin to delay
starch gelatinisation during heating. Experimental evidence shows that the volume expansion
ratios of dough depend on DF type (Fig. 1).
3,5 3,5

leavening baking leavening baking

3,0 3,0

2,5 2,5

V/Vo
V/Vo

2,0 2,0

1,5 1,5

a) b)
1,0 1,0
0 15 30 45 60 75 90 105 0 15 30 45 60 75 90 105
Time (min) Time (min)

3,5

leavening baking
3,0

2,5
V/Vo

2,0

1,5

c)
1,0
0 15 30 45 60 75 90 105
Time (min)

Figure 1. Variation in the volume ratio of: a) control; b) LI and c) CS during leavening and baking.

A negative effect of DFs on volume ratio is observed during breadmaking. LI dough shows the
minimum increment volume during leavening, with the fermentation time being shorter. The
highest volume increase of the baked bread corresponds to the control dough.

CONCLUSION
Results both dough rheology and bread properties drive on to conclude that the functional role
of a high amount of DF on structure formation in bread during baking process greatly depends
on fibre types.

REFERENCES
[1] Steffe J.F. 1992. Rheological methods in food process engineering. In Freeman Press, USA. pp 258-
263.

2070
Extraction techniques of red and green propolis: extraction yield of phenolic compounds

Losiane Paviania, Patricia Sacodaa, Erika Saitoa, Fernando Cabrala

a
Departament of Food Engineering, State University of Campinas, Campinas, Brazil
(cabral@fea.unicamp.br)

INTRODUCTION
Propolis possesses antibacterial, antifungal and antiviral properties and many other beneficial
biological activities: anti-inflammatory, antiulcer, local anaesthetic, hepatoprotective,
antitumos, immunostimulating [1], anti-HIV, among others. The various biological activities of
propolis have been attributed mainly to the presence of phenolic compounds, especially
flavonoids and phenolic acids. Solvents with different polarities (hexane, ethyl acetate, ethanol
and water) were used for the soxhlet extraction and the supercritical assays were carried out
using the dynamic method to fractionate the dry EEP of red propolis. The supercritical extracts
were obtained at temperatures of 40, 50 and 60 °C and pressures of 300 and 400 bar. All
extracts were analyzed for total phenols and flavonoids. The supercritical fluid extraction using
CO2 as solvent has a high selectivity for the fractionation, mainly in operational condition of
higher temperature and pressure studied.

MATERIALS & METHODS

Samples of red propolis were obtained at Ilha do Porto Apiary (Marechal Deodoro, AL).
Samples of green propolis, native to the State of Minas Gerais, Brazil, classified as group 12
for Park et al. [2] (Brazil has 13 different groups of propolis, with distinct characteristics), were
obtained from Bioessens Ltda. (Cotia, São Paulo, Brazil). Soxhlet extraction for green propolis
were caried out using ethanol, ethyl acetate, n-hexane and distilled water. The experimental
apparatus and procedure for SFE have been described in detail in other work [3]. The
operational conditions for SFE with CO2 for red propolis were: 40, 50 and 60 °C and from 300
to 400 bar. The The total polyphenol content and total flavonoids were determined in the
extracts.

RESULTS & DISCUSSION

The results of extraction yield (in %) are shown in Table 1 for low pressure extraction
methods: EEP and Soxhlet with different solvents for green propolis: Hex, EtAc, EtOH, H2O
and SFE. The largest yield was obtained by Sox-EtOH (49.38 ± 1.37) and Sox-EtAc (45.90 ±
1.41), solvents with intermediate polarity. The highest yield obtained by SFE was about 13% at
60°C and 300 bar, which is much lower than Xo by Sox-EtOH (49.38 ± 1.37% w/w). In
general, the extraction yields increase with increasing temperature and pressure. The extraction
of phenolic compounds from EEP was more efficient than those obtained by supercritical fluid
extraction, except for the 60°C and 400 bar. For the flavonoids, the EEP is lower than those
found in supercritical extraction in all conditions analyzed.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2071
 Table 1. Global yield (X0 % w/w) of propolis extract obtained by conventional techniques and
supercritical fluid extraction (SFE).
Extraction method Solvent Solvent Xo Total Total
polarity index (% w/w) polyphenolics flavonoids
(mg GAE/g) (mg CE/g)
EEP- Green propolis EtOH 5.2 36.78 138.59 36.78
EEP- Red propolis EtOH 5.2 45.00 215.44 45.00
Sox- Green propolis Hex 0.0 10.52 ± 1.18 93.75 10.52
Sox- Green propolis EtAc 4.4 45.90 ± 1.41 192.68 45.90
Sox- Green propolis EtOH 5.2 49.38 ± 1.37 147.39 49.38
Sox- Green propolis Water 9.0 15.75 ± 0.99 94.96 15.75
SFE - 40 °C/ 300 bar CO2 3.56 147.27 3.56
SFE - 40 °C / 400 bar CO2 5.00 116.19 5.00
SFE - 50 °C / 300 bar CO2 6.57 114.48 6.57
SFE - 50 °C / 400 bar CO2 9.74 142.72 9.74
SFE - 60 °C / 300 bar CO2 12.86 117.46 12.86
SFE - 60 °C / 400 bar CO2 12.26 229.90 12.26

CONCLUSION
The results of the extraction of raw green and red propolis different solvents showed high
extraction yields, especially when ethyl acetate and ethanol were used as solvent. The phenolic
content was higher in the extraction of red propolis (EEP-Red propolis) than green propolis
(EEP-Green propolis). All supercritical extracts presented higher flavonoids concentration to
the initial value found in EEP, indicating that SFE has a tendency to focus on flavonoids,
which makes supercritical fluid extraction very interesting to fractionate compounds of
propolis, which according to several authors, have higher biological activity.

REFERENCES
[1] Burdock G. A.. 1998. Review of the biological properties and toxicity of bee propolis (propolis).
Food and Chemical Toxicology, 36, 347–363.
[2] Park Y. K., Alencar S. M., Aguiar C. L. 2002. Botanical origin and chemical composition of Brazilian
propolis, Journal of Agricultural and Food Chemistry, 50, 2502-2506.
[3] Martinez-Correa H. A, Magalhães P. M., Queiroga C. L., Peixoto C. A., Oliveira, A. L., Cabral F. A.
(2011). Extracts from pitanga (Eugenia Uniflora L.) leaves: Influence of extraction process on
antioxidant properties and yield of phenolic compounds. The Journal of Supercritical Fluids, 55, 998-
1006.

2072
 Correlation of hydro-thermal processing with rutin content in tartary buckwheat flour

Jiyoung Yooa, Seung Mi Leea, Soojung Heoa, Sang-Ho Yooa, and Suyong Leea

a
Department of Food Science and Technology, Carbohydrate Bioproduct Research Center, Sejong
University, 98 Gunja-dong, Gwangjin-gu, Seoul 143-747, Republic of Korea (suyonglee@sejong.ac.kr)

INTRODUCTION
Recently, buckwheat (Fagopyrum spp) that belongs to the family Polygonaceae has been
receiving great attentions as an alternative crop due to its nutritional superiority to cereals.
Especially, rutin that is the primary phenolic compound of buckwheat, is shown to reduce the
risk of arteriosclerosis and high blood pressure, antagonize the increase of capillary fragility
associated with hemorrhagic disease. It is however interesting to note that the reduction of rutin
content is generally observed when buckwheat seeds are ground into flour or mixed with water
due to rutin-degrading enzymes (RDEs) [1]. Therefore, the reduced amount of rutin as well as
bitter taste may play a negative role in consumer preferences, consequently discouraging the
food industry to develop a variety of buckwheat-based foods. Thus, the goals of this study were
to apply several hydro-thermal processing into buckwheat flour and to establish the
experimental procedures to minimize the rutin loss during buckwheat processing.

MATERIALS & METHODS

Tartary buckwheat (F. tataricum) flour (40 g) was subjected to three different hydro-thermal
treatments – steaming, autoclaving, and boiling. For steaming, buckwheat flour was placed on
a plate in a steam cooker with a lid and steamed over boiling water for 10 min. In case of
autoclaving, buckwheat flour was placed into an autoclave for 10 min at 120ఁ. Also,
buckwheat flour was immersed in boiling water for 10 min and then freeze-dried. Raw and
hydro-thermally treated buckwheat (6 g) samples were mixed with distilled water (4 ml) for 0,
2, 5, 10, 30, 60 min and freeze-dried. Buckwheat flour (1 g) was treated with 20 ml of
methanol at 80Ȕfollowed by cooling overnight at 4Ȕ After filtering through Whatman filter
paper (No. 41) and 0.45 ห filter paper, the amounts of rutin and quercetin were quantitatively
measured by using HPLC (Agilent, Santa Clara, CA, U.S.A.) with a UV detector (350 nm) and
Capcell Pak C18 column[2]. The mobile phase for HPLC consisted of methanol/ acetic acid
(95: 5, v/v) (solvent A) and water (solvent B). A linear gradient of the solvent A was applied
from 10% to 60% for 40 min, followed by an increase to 100% in 5 min.

RESULTS & DISCUSSION

The contents of rutin and quercetin in steamed buckwheat flour were measured and compared
with those of raw buckwheat flour. As shown in Fig.1, 46.06 mg/g of rutin was contained in
raw buckwheat flour while the rutin content dramatically decreased when the buckwheat flour
was mixed with distilled water. On the other hand, the quercetin content significantly increased
from 0.52 mg/g to 31.2 mg/g. Therefore, it seemed that mixing with water caused rutin-
degrading enzymes in buckwheat flour to easily be accessible to rutin, which consequently was
degraded into quercetin These results are in a good agreement with those reported by Suzuki et
al [3]. However, as also can be seen in Fig. 1, the rutin loss by the addition of water was not
observed in the steamed buckwheat flour and also quercetin was hardly detected.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2073
Figure 1. Effect of mixing time with water on the rutin (a) and quercetin (b) contents of raw and steamed
buckwheat flours

The contents of rutin and quercetin in autoclaved buckwheat flour were investigated. The
autoclaved buckwheat samples contained 45.27 mg/g of rutin and 0.7 mg/g of quercetin which
were similar to those of the control before mixing with water. It thus seemed that autoclaving
caused rutin in buckwheat flour to remain constant even though mixed with water. The
amounts of rutin and quercetin in boiled buckwheat flour were investigated. Boiling treatment
also resulted in the similar pattern of rutin content in buckwheat flour to the steaming and
autoclaving treatments. However, the content of quercetin in boiled buckwheat flour (2.48
mg/g) was 3 to 4 fold higher than that of two other hydro-thermally treated samples, which was
however not varied over mixing times. The results indicate that the rutin-degrading enzyme
became deactivated by hydro-thermal treatments, consequently preventing the rutin loss in
buckwheat flour by the addition of water.

CONCLUSION
In this study, tartary buckwheat flour was subjected to three different hydro-thermal processing
in order to prevent rutin loss by rutin-degrading enzymes. The addition of water to raw
buckwheat flour led to a dramatic decrease in rutin content and increase in quercetin content
probably since it made rutin susceptible to the attack of rutin-degrading enzymes. However, the
rutin contents in buckwheat flour after steaming, autoclaving, and boiling were not changed by
water addition, which also did not produce quercetin. Therefore, the use of hydro-thermally
treated buckwheat flour can provide more health benefits derived from a high amount of rutin,
probably encouraging the food industry to develop various buckwheat-based food products.

REFERENCES
[1] Yasuda T. & Nakagawa H. 1994. Purification and Characterization of the Rutin-Degrading Enzymes
in Tartary Buckwheat Seeds. Phytochemistry, 37(1), 133-136.
[2] Terada H. & Miyabe M. 1993. Determination of Rutin and Quercetin in Processed Foods by Fast
Semi-Micro High Performance Liquid Chromatography. Journal of the Food Hygienic Society of
Japan, 34(5), 385-391.
[3] Suzuki T.,Honda Y.,Funatsuki W. & Nakatsuka K. 2002. Purification and Characterization of Flavonol
3-Glucosidase, and Its Activity During Ripening in Tartary Buckwheat Seeds. Plant Science, 163(3),
417-423.

2074
 Study of the influence of berry-blanching on syneresis in blueberry purées

Ada Brambillaa, Dario Maffib, Anna Rizzoloa

a
Consiglio per la Ricerca e Sperimentazione in Agricoltura, Unità di ricerca per i processi dell’industria
agroalimentare (CRA-IAA), Milan, Italy (adaelisabetta.brambilla@entecra.it; anna.rizzolo@entecra.it)
b
Università degli Studi di Milano, Dipartimento di Produzione Vegetale, Sezione di Patologia Vegetale,
Milan, Italy (dario.maffi@unimi.it)

INTRODUCTION
The need to supply convenience food preserving health-related compounds, moves research
towards the application of mild technologies and the introduction of innovative solutions for
the consumer. Frozen blueberry purée, processed without pomace loss, is a versatile product
potentially rich in all the bioactive compounds characteristic of the whole berry. The ability of
a steam blanching pre-treatment to improve anthocyanin yield and recovery from pomace has
been assessed in blueberry juice and it has been related to thermal inactivation of fruit
oxidative enzymes [1] and to structural alterations of pigmented epidermal cells. This work
aimed at studying the influence of blueberry blanching pretreatment on the kinetic of frozen
purée syneresis and on colour parameters and phenolic composition of the separated serum.
Correlation between thermal treatment and tissue localization of pigments in whole berries was
highlighted by macro- and micrographs and implications on purée quality attributes discussed.

MATERIALS & METHODS

Highbush blueberries cv Brigitta were IQF and stored at –20°C till processing and analyses.
Before puréeing, one half of the berries was thawed at 20°C for 3h (NB), while the other half
was steam-blanched for 3min and tap water-cooled in a pilot steam blanching tunnel (BL).
Then 300g aliquots of purées were packed in plastic vessels, sealed under partial vacuum,
frozen and stored at í20°C up to analyses. Formic acid extracts [2] of berries and purées were
prepared and frozen (í20°C) till phenolics analysis. NB and BL berries were evaluated for
monomeric anthocyanin pigments (MAP), total phenolic compounds (TPC), percent polymeric
colour (%PC), index of browning (IB) and for macro- and micrographs on half berries using a
digital camera and on berry sections by light microscope. On frozen NB and BL purées the
syneresis kinetics have been studied at 20°C for 24h, weighing the juice from syneresis at 30
min intervals up to 6 hours upon thawing, and then after 24h. A model fitting weight data was
elaborated using the Simple Regression procedure (Statgraphics). NB and BL purées thawed
overnight (4°C) and collected juice from syneresis were analysed for MAP, TPC, %PC and IB,
and only juice for colour parameters.

RESULTS & DISCUSSION

Steam blanching induced a marked anthocyanin diffusion from pigmented epidermal layers
down to the core of berries, due to thermal induced plasmolysis and wall softening phenomena
(Figure 1). Pigments flow was correlated to a decrease in MAP in BL berries (í17.6%), due to
a direct heat-induced damage on berry pigments. Nevertheless, BL purées achieved a higher
content in MAP (91.7 vs. 82.4 mg/100g) and TPC (276.9 vs. 182 7 mg/100g) and a lower %PC
compared to NB purées, evidencing a protective effect of blanching along the purée processing

11th International Congress on Engineering and Food - Athens, Greece, 2011 2075
chain. Liquid-holding capacity was higher in BL purées during the first 180 min after thawing,
then it was higher in NB ones till 340 min of time. The best model fitting both BL and NB
syneresis data is reported in Eq. 1.
W ab t (1)
where W is the weight of serum, t is the syneresis time, and a and b are the estimated
parameters of the model. Although the NB and BL serum weights were the same after 24 h at
20°C, BL samples had a higher content in TPC (175.9 vs. 104.0 mg/100ml) and triple the
amount in MAP compared to NB samples; in addition BL syneresis liquids had higher
lightness (L*) and red colour component (a*). As a consequence, a strong phenolic-enriching
effect of blanching on the liquid portion of blueberry purées, especially concerning
anthocyanin compounds, can be argued. This effect, together with heat solubilisation of tissue
pectins, could account for the different liquid-holding capacity of BL and NB purées.

Figure 1. Effect of steam blanching on pigments diffusion in V. corymbosum L. (A, not blanched; B,
blanched). 1-Berries longitudinal sections. 2-Epidermis panoramic views. 3-Epidermis cross-sections
(Bars=30 μm).

CONCLUSION
The introduction of a berry-blanching step in the processing of ready-to-eat frozen blueberry
purées would enhance nutritional and sensory qualities, preserving phenolic compounds,
limiting syneresis phenomena and improving colour properties. Enriching effect of blanching
on the liquid portion of blueberry purées, supported by biochemical and microscopy evidences,
may have further nutritional and technological implications to be better investigated.

REFERENCES
[1] Rossi M., Giussani E., Morelli R., Lo Scalzo R., Nani R.C. & Torreggiani D. 2003. Food Research
International, 36, 999-1005.
[2] Brambilla A.; Lo Scalzo R.; Bertolo G.; Torreggiani D. 2008. Journal of Agricultural and Food
Chemistry, 56 , 2643-2648.

2076
 Quality properties of corn-based extrudates enriched with dietary fibers

A. N. Gianninia, M. K. Krokidaa, N. P. Zogzasb

a
Laboratory of Process Analysis and Design, School of Chemical Engineering, National Technical
University of Athens, 5 Iroon Polytechniou St., Zografou Campus, 15780 Athens, Greece,
e-mail: mkrok@ chemeng.ntua.gr
b
Laboratory of Food Engineering, Department of Food Technology, Technological Educational Institute
(TEI) of Athens, Agiou Spyridonos St., 122 10, Egaleo, Athens, Greece, e-mail: nzogzas@ teiath.gr

INTRODUCTION
Dietary fiber-rich materials have gained popularity as food ingredients for health benefits in
recent years [1]. However, relatively little is known about the effects of processing conditions
on structural properties of fiber containing foods. Extrusion is a unique and versatile thermal
process, where the raw materials are subjected to direct mixing and cooking, resulting to ready
to eat expanded snacks. The incorporation of dietary fibers into extruded products plays an
important role on their structural characteristics. High levels of fiber have often resulted in a
compact, tough, non-crisp and undesirable texture [2]. The purpose of this study was the
investigation of the effect of process conditions (temperature, feed rate) and raw material
characteristics (moisture content, fiber to corn ratio) on structural and mechanical properties of
corn based extrudates enriched with apple and oat fibers. Simple power models were
developed, to predict porosity and stress of compression as functions of process conditions and
material characteristics.

MATERIALS & METHODS

Oat and apple fibers were mixed with corn flour, with fiber to corn ratio ranging from 10 to
30%. The moisture content of the feed was adjusted from 13 to 19% wet basis, by adding the
necessary distilled water. A prism Eurolab conical, counter-rotating twin screw extruder, model
KX-16HC, with a process rotation speed of 200 rpm was used. The screw geometry was:
length 40 cm, diameter 16 mm and die diameter 3 mm. Extrusion temperatures and mass rates
of feed were regulated from 150 to 230ºC and 0.5 to 2.0 g/s respectively. Apparent density,
true density and porosity of extrudates were determined by measuring the actual dimensions of
the samples, along with their true volume, using a Quantacrome stereopycnometer (model
SPV-3) with ±0.001 cm3 accuracy. Expansion ratio was calculated by the extrudate’s over the
die’s diameter and mechanical properties were evaluated through compression tests using a
Zwick Universal Testing Machine 1120.

RESULTS & DISCUSSION

The following parametric model was proposed to predict porosity as a function of process
conditions and material composition [3]:
nT nF nX nC
§T· §F· § X · § C·
¨ ¸ ¨
H Ho ˜ ¨ ¸ ˜ ¨ ¸ ˜ ¨ ¸ ¨ ¸ ¨
¸ ˜¨ C ¸ ¸ (1)
© Tr ¹ © Fr ¹ © X r ¹ © r¹

11th International Congress on Engineering and Food - Athens, Greece, 2011 2077
Where, İ is the porosity of the extrudate, T is the extrusion temperature (ºC), F is the feed rate
(gr/s), X is the feed moisture content (kg/100kg wb) and C is the materials ratio (kg fiber/kg
corn), Tr, Fr, Xr and Cr are their reference values and İȠ, nT, nF, nX, nC, are the respective five
parameters that were evaluated through regression analysis.
The mechanical stress of compression was modeled by the following equation:
p
V E ˜ e  (Vmax  E ˜ e max )(e / emax ) (2)

Where, ı is the stress of compression and ımax the disruption stress in (kPa), e is the strain
(ǻl/lo) and emax the disruption strain, Ǽ is the elasticity parameter in (kPa) and p is the
viscoelastic exponent. The values of E, ımax and emax can be related to process conditions and
material composition by using parametric models similar to that of equation (1).
The estimated values of porosity parameters for both types of extrudates are shown in table 1,
where SR and SE are the standard deviation and standard error respectively.

Table 1. Results of Parameters’ Estimation

Extrudate İo nT nF nX nC SR SE
Corn/apple fiber 0.92 -0.31 0.097 -0.16 -0.085 0.032 0.039
Corn/oat fiber 0.87 -0.13 0.02 -0.17 -0.065 0.032 0.047

As it can be seen from the above table and equation (1), the influence of feed rate, as it is
expressed by the exponent nF, is positive for both types of extrudates. That is, the extrudates’
porosity increases with the increase of feed rate. On the contrary, the negative values of nT, nX
and nC indicate the opposite behavior. That is, the increase of temperature, initial material
moisture content and fiber to corn ratio, leads to lower porosity values. Lower porosity values
have also been observed for the case of oat fiber to corn extrudates, something which is
justified by the lower value found for İȠ. As far as the stress of compression is concerned, it
was found that the influence of process conditions and material composition had the opposite
effects from those of porosity, in contrast with the expansion ratio that revealed the same
behavior. Similar results to this work have also been reported by other researchers [2].

CONCLUSION
Structural and mechanical properties of expanded corn-fiber snacks, produced on a twin screw
extruder, depend on several process conditions (extrusion temperature, feed rate, feed moisture
content and corn to fiber ratio). The addition of dietary fibers in feed results to more dense
products, while the use of apple fibers, leads to products with greater porosity and lower
compression stress values compared to those of oat fibers. It is possible to predict porosity and
mechanical properties of extrudates as functions of process and material composition variables.

REFERENCES
[1] Trowell, H., Burkitt, D. & Heaton, K. eds. 1985. Dietary fiber, fiber-depleted foods and disease.
Academic Press, New York.
[2] Lue, S., Hsieh, F., Huff, H.E., 1991. Extrusion Cooking of Corn Meal and Sugar Beet Fiber: Effects
on Expansion Properties, Starch Gelatinization and Dietary Fiber Content, Cereal Chem, 68, 227-234.
[3] Krokida, M.K., Zogzas N.P., Maroulis Z.B., 1997. Modelling Shrinkage and Porosity during Vacuum
Dehydration. Int. J. Food Sci. Technol., 32, 445–458.

2078
 Processing of berries

Ingegerd Sjöholma, Jitmanee Pullawanb, Marilyn Raynerc

Lund University, Department of Food Technology, Engineering and Nutrition, Lund, Sweden
a
(ingegerd.sjoholm@food.lth.se), b(jitmanee.pullawan.935@student.lu.se),c(marilyn.rainer@food.lth.se)

INTRODUCTION
Fresh berries have often a very short shelf life and are preserved by freezing, drying, and heat
treatment and often with some additional sugar. One way to increase the utilization of berries is
to start from frozen berries, thaw them and then use a combined osmotic treatment and
convection drying procedure. To describe the quality of these processed berries is a challenge
as the water content will be of economical interest and will also influence the texture and the
water activity will decide the safety and the storability of the product. One way to describe and
be able to predict these quality parameters of the final product is to measure the moisture
sorption isotherm, MSI of the product. MSI of each food is unique and it depends on
microstructure of the food, physical chemical state of the food components and composition
[1]. Many model equations are described in literature [2]. The aim of this study was to screen
different levels of pre-treatments, osmotic treatment and convection drying procedures to
obtain data to develop sorption isotherms for different berries.

MATERIALS & METHODS

Trials are performed on commercial frozen strawberries (Möllers, Denmark), blueberries
(Willys, Axfood), raspberries (Polarica Sweden) and blackcurrants (Bri9568-10). Each berry is
selected to have similar size, measured by length or diameter. Strawberries, 3-4 cm,
blackcurrants, 1-1.5 cm, blueberries, 0.5 to 0.8 cm and raspberries 1.7-2.1 cm. Granular sugar
(Dan sukker, Danisco Sugar) is used as osmotic medium in all trials. Osmotic treatment of
berries is done in closed glass jars in room temperature (20 ºC), each batch around 50 grams.
Sugar amount in osmotic treatment are varied from berries per sugar weight 2:1(33% sugar),
3:1(25% sugar) and 4:1(20% sugar). Blackcurrants and strawberries are used in this trial to
represent berry samples. Frozen blackcurrants are steamed 90 seconds, strawberries are cut into
halves.
The efficiency of osmotic treatment process is evaluated by water loss (WL), solid gain (SG)
and mass reduction (MR). Calculation of MR, WL and SG used equations below.
MR = 100- M ; WL = (100* b1) – M * b2 ; SG = WL – MR Where M is weight after
osmotic treated (based on 100 g initial weight), b1 is moisture content of fresh berries /100 and
b2 is moisture content after osmotic treated /100 [3].
The osmotic treated berries are dried in a convection oven, 3 m/s and 60ºC. Temperatures and
weight are monitored continuously during the drying operation. The osmotic treated berries are
dried until different levels of water content. Water content of the dried samples are measured in
a vacuum oven, 60 ºC until constant weight and the water activity is measured in a Aqualab T2
set up. The different moisture sorption isotherms of the dried berries were fitted into different
models [2].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2079
RESULTS & DISCUSSION
The results showed that pre-treatment is not needed for strawberries, raspberries and
blueberries; however blackcurrants require a 90 second steam treatment before the osmotic
treatment step. Whole strawberries were cut in halves in order to shorten the convective drying
time. The used osmotic treatment conditions, granular sugar ratio berries per sugar
4:1(20%sugar) for 24 hours were suitable for all 4 berries. Moisture sorption isotherms were
generated based on dry matter and water activity of each berry. The moisture sorption isotherm
of the dried berries were tested in the different models; Halsey's equation, GAB (Guggenheim-
Anderson-de Boer) equation, Henderson's equation respectively Kuhn equation by using
statistical software [2]. The fitting showed that GAB(Guggenheim-Anderson-de Boer) equation
fits the best to the individual isotherms and can be used to predict equilibrium moisture of all
four berries when water activity aw is in the range up to 0.94, see table 1.

Table 1. GAB constant parameters for each berry

Berries Constant Parameters

Į ȕ Ȗ R2

Raspberries 13,3376 -26,4919 13,8101 0,9623

Blueberries -5,1126 2,38279 2,8885 0,9708
Strawberries 10,0876 -21,5269 12,0734 0,9096
Blackcurrants 17,0593 -31,9357 15,4811 0,9871

CONCLUSION
The moisture sorption isotherm can be of benefit for food producers to design their drying
processes to meet the moisture conditions in the final product and give the possibility to use
these healthy and natural ingredients in a wider range of food products.

REFERENCES
[1] Rockland A.B., Beuchat L.R.(1987). Water activity : Theory and applications to food Marcel
Dekker, New York, pp.215-231.
[2] Heldman D.R.& Lund D.B. (1992). Handbook of Food Engineering. Marcel Dekker, New York,
pp.437-562.
[3] Welti Chanes J.,Velez-Ruiz J.F., Barbosa Canovas G.V. (2003). Transport Phenomena in Food
Processing. CRC Press. pp.83-94.

2080
 Aroma Release and Sensory Perception of Fruit Candies Model Systems

Piccone Pierpaolo a, Rastelli Simon Luca b, Paola Pittiaa,

a
Department of Food Science, University of Teramo Mosciano S.Angelo (TE), Italy (ppittia@unite.it)
b
Gelco s.r.l. (Perfetti van Melle Group), Castelnuovo Vomano (TE), Italy

INTRODUCTION
The release of volatile compounds from food matrices is governed by kinetic and thermodynamic
phenomena [1]. The rate of release of volatiles and their partitioning in the gas phase is influenced by
intrinsic, extrinsic factors as well as the interaction with non volatile compounds in the food
matrix..Gummy candies are confectionery products and their texture is achieved by using various gelling
agents; the most important ingredients are the sweeteners i.e. sucrose, glucose and corn syrups. The
objective of this study was to investigate the influence of the composition (i.e. gelling agents - gelatine,
starch pectin- and sugars) on the release and sensory perception of aroma compounds from candy model
systems added with a standard strawberry flavour.

MATERIALS & METHODS

The following model candies were prepared following recipes mimicking those used for commercial
products: gelatine (G), aerated gelatine (GA), gummy gelatine (Ggo), gelatine and starch (GA), extruded
starch (Aes), pectin (P). All samples were added with the same concentration of a strawberry aroma
(0.25% db) and with the exception of Aes, all models, after setting, were dried under the same process
conditions and stored in HDPE bags until analysis. Evaluations of the release of aroma compounds under
equilibrium conditions were carried out by Head Space Gas Chromatography both on the model candies
and the same submerged in water to mimic the dissolution in the mouth. In the latter case, the kinetics of
the aroma release from the models during dissolution to reach the equilibrium were also investigated.
Electronic nose (EN) measurements and sensory evaluations under standardised procedure were also
carried out.

RESULTS & DISCUSSION

The candy model systems differently prepared are characterised by a moisture content in the range of 16.9
%(Gar) and 23.2 % (GA), whilst Aes, due to the preparation process showed the lowest moisture content
(12%). Different were also the textural properties (compression and penetration test) being GA the more
firm and tough. Among the 16 volatiles of the strawberry flavour, 4 out of them were selected (ethyl
hexanoate-EH, ethyl butyrate-EB, methyl cinnamate-MC, ethyl acetate-EA) to represent volatiles with
different polarity. The partitioning of strawberry aroma compounds from the candy model systems at
equilibrium in the head space was highly affected by the gelling agent type and sugar content. In Figure 1,
the ratio of the GC area of EH and EA in the model systems to the GC area of the same compounds in the
pure strawberry aroma is reported and considered as index of the volatile retention. The affinity of the
aroma compound for the biopolymer/s of the candy model system and structural properties are implied in
the different release in the vapour phase of EA (more polar) and EH (more apolar). The kinetics of the
volatiles release from the candies in water varied depending on the aroma nature and the physical
properties of the systems as affected by the gelling agent. In particular a lower aroma release rate
occurred for all the aroma compounds in the starch-gelatine model systems while the highest occurred in
pectin- and gelatine- based candies. The latter results are in agreement with the sensory analysis results

11th International Congress on Engineering and Food - Athens, Greece, 2011 2081
that showed a higher intensity of the pectin made samples upon dissolution and chewing. EN analysis
discriminated samples based on composition and sugars content.

0.30

EA
0.25
Acaramella/Aaromapuro

0.20

0.15

0.10

0.05

0.00
P G Gar Ggo GA Aes
Modelsystem

0.30 Figure 1. Ratio of the

EH GC area of EH and EA in
0.25 the differently prepared
candy model systems to
Acaramella/Aaromapuro

0.20 the GC area of the same

compounds (EA, EH) in
0.15 the pure strawberry
aroma.
0.10

0.05 CONCLUSION
Composition, type of
0.00
P G Gar Ggo GA Aes biopolymers and physical
ModelSystem properties affect the
retention of the aroma
compounds in the candies and their release when dissolved in water..

REFERENCES
[1] Voilley A, Souchon I (2006). Flavour retention and release from food matrix: an overview. Flavour in foods.
Cambridge: Woodhead Publishing Ltd, 31, 305 – 316

2082
11th International Congress on Engineering and Food - Athens, Greece, 2011 2083
fermented sausages was detected by Muthukumarasamy et al. [3]. S. xylosus numbers
maintained on the level of 6 log cfug-1 during 14 days of drying, and during the storage they
increased to the level of about 1 log cfug-1 in all treatments. Similar trends were detected in dry
fermented sausages produced with S. carnosus [3].

Figure 1. Survival of probiotic bacteria during ripening and storage of dry fermented sausages

In both variants (2, 3) of probiotic sausages, the initial counts of probiotic bacteria were
approximately 6 log cfug-1 (Fig.1). During 3 days of sausage drying, the probiotic bacteria
counts increased by 2 log units and maintained on the level of 8 log cfug-1 until the end of the
storage period. Fermented sausages were sensory evaluated on days 14 and 40. After 14 days,
all quality parameters were evaluated relatively high, with grades over 7, without significant
differences between sausage variants. After 40 days, variants 1 and 2 were very similar with
grades over 7.6, but variant 3 received somewhat lower grades for aroma (6.75), taste (6.0) and
texture (6.25). All samples were evaluated as acceptable.

CONCLUSION
It can be concluded that probiotic Lactobacillus helveticus RO52 and Bifidobacterium longum
RO175 (Lallemand, France) seem to be suitable for the production of dry sausages. The
survival of probiotic bacteria was on a high level of 8 log cfug-1 at the end of storage period in
both probiotic sausage variants. Chemical composition and pH values were similar in control
and probiotic samples of sausages. Sensory quality was acceptable in all sausage variants, but
somewhat better aroma, taste and texture were detected in sausages produced with
Lactobacillus helveticus RO52.

REFERENCES
[1] Erkkilä S., Suihko M.L., Eerola S., Petäjä E., Mattila-Sandholm S. 2001. Dry Sausage Fermented by
Lactobacillus rhamnosus strains. International Journal of Food Microbiology, 64, 205-210.
[2] Klingberg T.D. & Budde B.B. 2006. The Survival and Persistence in the Human Gastrointestinal
Tract of Five Potential Probiotic Lactobacilli Consumed as Friez-Dried Culture or as Probiotic
Sausage. International Journal of Food Microbilogy, 109, 157-159.
[3] Muthukumarasamy P. & Holley R.A. 2006. Microbiological and Sensory Quality of Dry Fermented
Sausages Containing Alginate-Microencapsulated Lactobacillus reuteri. International Journal of food
Microbilogy, 111, 164-169.

2084
 Evaluation of probiotic containing microcapsules stability in different media

Luciano Avallone Buenoa*, Maria de Fátima Fonsecab, Djalma Marquesb, Fernanda Branco Shinagawab,
Amanda Quintinob, Gabriel Locatellib, Cedenir Pereira Quadrosb

a
University Rural of Pernambuco, Physical Department, Recife, Brazil; *(avallonebueno@gmail.com)
b
BioLogicus – Research Center of Probiotics, Technology Institute of Pernambuco, Recife,
Brazil;(www.biologicus.com.br)

INTRODUCTION
Probiotics are viable microorganisms preparation which have beneficial effects on the host
health. Various health benefits have been attributed to probiotics such as antimutagenic and
anticarcinogenic properties, antiinfection, immune system stimulation, lactose intolerance.
Successful probiotic microorganisms are able to survive in gastric conditions and colonize the
intestine by adhering to the intestinal epithelium [1]. Microencapsulation of microorganism is
one of the newest and most efficient methods, has recently been under especial consideration
and investigation. Alginate is often used as an encapsulating material because it has the
benefits of being non-toxic and being readily available. Chitosan polymers can further
polymerize by means of cross-link formation in the presence of anions and polyanions [2]. The
aim of this work was to evaluate the influence of substances polymeric on the physicochemical
stability of microencapsulated microorganisms stored in two kinds of media, as well as the
microencapsulated resistance against treatment with different ranges of pH.

MATERIALS & METHODS

The process of microencapsulation was carried out by method of coacervation which was used
as encapsulate material the fermented soybean beverage BioLogicus® as probiotic
microorganism. After this, were resuspended in mineral water and 0.85% saline solution and
stored at 5ºC until analysis. For stability tests, pH, acidy evaluation, % mass variation and total
soluble solid was analysed, during 84 days. For the resistance test, 10 g of alginate-chitosan
free were immersed in 100 mL of acid solution containing KCl buffer pH 2.0 for 3 h at 37°C.
For alkaline system, was used KH2PO4 pH 7.4 for 3 h at 37°C.

RESULTS & DISCUSSION

Generally, total titratable acidity increased in two storage media which were submitted the two
types of encapsulation. In contrast to the acidity, pH profile decreased during the 84 days.
Media composed by 0.85% saline solution, that stored the alginate-chitosan and alginate-
chitosan free microcapsules, shows an upward trend to titratable acidity and decreasing for pH
measure. On the other hand, the two media, which stored alginate-chitosan microcapsules, had
the titratable acidity variation since the first week. The decrease of the pH is explained due to
the presence of substrate that acts as means of development of microorganism. Results of these
reactions were expected considering that the array can be used as the substrate for development
of microorganisms, which as a result of metabolism, producing lactic acid. In relation to total
soluble solids variation in storage media, the variation was very insignificant, around of the 0.1
a 0.2°Brix. Although the values were relatively constant, it was seen a higher initial value for
0.85% saline solution assay. For mass variation determination, were weighed in the same time

11th International Congress on Engineering and Food - Athens, Greece, 2011 2085
interval, four aliquots stored in mineral water or 0.85% saline solution. It was observed that, in
the first week, alginate-chitosan free obtained a mass gain of the 8 and 29% for mineral water
and saline solution respectively. Result was very similar with saline solution medium, but in
case for alginate-chitosan microcapsules the mass gain was 16%. The acid aqueous solution
remained clear during all time of the analysis. This is suggests that microcapsules are resistant
to gastric juice. However, the microcapsules that were submitted to alkaline treatment,
demonstrated a complete dissolution after 3 h. It can be noted the dissolution process of the
microcapsules in acid and alkaline solution. These results demonstrate that alginate and
alginate-chitosan microcapsules have an ideal behaviour in the human organisms.

A
B

C
D

Figure 1. (A) Matrix alginate-chitosan free microcapsules, (B) matrix alginate-chitosan microcapsules on
mineral water medium and (C) matrix alginate-chitosan free microcapsules, (D) matrix alginate-chitosan
microcapsules on 0.85% saline solution during 84 days.

CONCLUSION
It was concluded that development of alginate-chitosan free and alginate-chitosan
microcapsules for probiotic microorganism immobilization can be storage both in mineral
water, as well as in 0.85% saline solution for long period. In addition, alginate microcapsules
show resistance to acid treatment and susceptibility to alkaline, ideal for liberation of the
bioactive agents in intestinal system.

REFERENCES
[1] Fuller R. 1991. Probiotics in human medicine. Gut 32(4), 439-442.
[2] Klien J., Stock J., Vorlop K.D. 1983. Pore size and properties of spherical calcium alginate
biocatalysts. European Journal Applied Microbiology Biotechnology, 18, 86-91.

2086
 Effect of Drying in Aloe’s Functional Components

Ȃ. Krokida1, A. Pappa2, M. Agalioti1

1
Laboratory of Process Analysis and Design, School of Chemical Engineering, National Technical University of
Athens, 9 Iroon Polytechniou St., Zografou Campus, 15780 Athens, Greece.
2
Laboratory of Analytical Chemistry, School of Chemical Engineering, National Technical University of Athens, 9
Iroon Polytechniou St., Zografou Campus, 15780 Athens, Greece.,
athpappa@chemeng.ntua.gr

INTRODUCTION
Aloe vera is a member of the Liliacea. Aloe is widely used as a natural treatment and alternatively
therapy for various types of diseases. The plant can be separated into two products: aloe latex , a bitter
yellow exudates from the pericyclic tubules in the outer skin of the leaves and aloe gel. The gel consists
primarily of water (>98%) and polysaccharides. Acemannan is considered the main functional component
of aloe vera and is composed of a long chain of acetylated mannose [1]. The potential use of aloe vera
products often involves some type of processing, including among others drying, resulting in powdered
samples, which can further mix with other ingredients to form a great variety of dietary, cosmetics and
pharmaceuticals products. However, food products are sensitive to drying temperatures, which can being
induce degradation (e.g. oxidation, loss of texture, functional properties etc). Vacuum and freeze drying
were also tested as drying methods to provide powder samples with the least deterioration The aim of this
study was to evaluate the effect of freeze drying, along with vacuum and air drying in the preservation of
some aloe’s functional substances, such as, polysaccharides and minerals.

MATERIALS & METHODS

Fresh aloe vera leaves, of between 30 and 50 cm length, corresponded to a 4-year old plant, obtained from
Peloponnisos district in Greece, were used as the raw material. Whole leaves were washed with distilled
water and the epidermis was separated from the parenchyma by a scalpel–shaped knife. The filets were
washed with distilled water to remove the exudates from their surfaces, diced to 1cm3 cubes and
dehydrated either in a coventional air drier at 70oC (AD samples) or in a vacuum drier at 0oC (VD
samples). Aloe filets were also freeze dried (FD sample), and used as reference samples in NMR and FT-
IR analyses. Gel was collected from the inner part of aloe leaf and frozen at -30oC for two days. Prior to
freeze drying it was treated with liquid nitrogen for 1 h. 1H NMR spectra at 300 MHz were recorded.
Approx. 10 mg of dried Aloe vera samples were dissolved in 2 ml of 99.9% deuterium oxide, and
transferred in NMR tubes. No internal shift standard was added. Fourier Transformed Infrared (FT-IR)
spectra were obtained, after preparing a KBr disc containing 2 mg of freeze, air and vacuum dried
samples. The single beam traversing each sample was rationed with the single beam of the corresponding
background. The inorganic residues of the dried samples, as well as of fresh filets were determined after
heating at 550oC overnight. Then the residues were dissolved in HNO3. Minerals K, Na, Ca, Mg were
determined using an atomic absorption spectrometer. Heavy metals like Pb, Cr, Ni, Mn, Fe and Cu were
determined by an Inductively Coupled Plasma Atomic Emission Spectrometer. The P content was
estimated using the phoshoro-vanadium-molybdenum complex at 466 nm by a portable spectrometer.

RESULTS & DISCUSSION

Acemannan is a linear polysaccharide composed by ȕ-(1ĺ4)-linked mannan partially acetylated in
positions 2,3 or 6. In a 1H NMR spectrum these acetyl groups generate a characteristic signal (2.00 to

11th International Congress on Engineering and Food - Athens, Greece, 2011 2087
2.26 ppm) that can be considered as the fingerprint of aloe vera [2]. In the 1H NMR of FD sample, a clear
signal in the above mentioned ppm range is presented, which shows that during freeze drying the features
of fresh gel concerning the acemannan are remained. On the contrary, in AD samples, a remarkable
decrease of the signal of acemannan is observed and this decrease is more intense during the air drying.
The absence of signal at 1.33 ppm is a negative indication for the formation of lactic acid, showing that
the air and vacuum processes do not induced enzymatic degradation into aloe vera gel. In Fig. 1 the FT-
IR spectra of FD, VD and AD samples are presented. FTIR spectra of VD and AD samples revealed
remarkable decreases of the bands of 1740 and 1250 cm-1, which correspond to the C=O and C-O-C
stretches of the acetyl groups, in comparison with FD samples.

Figure 1. FT-IR spectra of freeze dried, vacuum dried, and air dried samples

The predominant mineral elements in fresh and dried samples are K, Na, Ca and Mg and secondly Mn,
Fe, Cu, Zn was in the range of a few mg/100 g dried material, d.m., whereas the toxic metals like Pb, Cr,
Cd was lower than 1 mg/100 g d.m. in all samples. The level of Ca was high in all samples. In
combination with other ions such as, Na, K, and Mg provides an ionic balance for the vascular membrane,
promoting vasorelaxation and a reduction in blood pressure [3]. The drying processes lead to small
differences into mineral content due to differences in solubility, through hardening process during drying.
Finally the P content was at low level unaffected from drying process.

CONCLUSIONS
Concentration of polysaccharides was affected at some extent from drying process, such as air and
vacuum drying, through detoration of acemannan polysaccharides as the NMR and FT-IR analyses
showed. On the other hand, minerals concentration remained practically constant.

REFERENCES
[1] J. H. Hamman, Composition and Applications of Aloe vera Leaf Gel, Molecules, 13 (2008) 1599-1612.
[2] A. Bozzi, C. Perrin, S. Austin, F. Arce Vera, Quality and authenticity of commercial aloe vera gel powders, Food
Chemistry, 103(2007) 22-30.
[3] M. Miranda, H. Maureira, K. Rodriguez, A. Vega-Galvez, Influence of temperature on the drying kinetics,
physicochemical properties and antioxidant capacity of Aloe Vera (Aloe Barbadensis Miller) gel, Journal of Food
Engineering. 91 (2009) 297-304.

2088
Functional foods enriched in aloe vera. Effects of vacuum impregnation and temperature

on the respiration rate and the respiratory quotient of some vegetables

Sigrid Sanzana a ; María Luisa Gras a ; Daniel Vidal-Brotóns a

a
Instituto Universitario de Ingeniería de Alimentos para el Desarrollo (IUIAD), Universidad Politécnica
de Valencia, Valencia, Spain (mgrasro@tal.upv.es)

INTRODUCTION
The inclusion of physiologically active natural components with beneficial effects on health
strengthens the nutritional value of fresh vegetables. Many scientific studies have been made
about aloe vera and have discovered some potential health benefits of its components. The
vacuum impregnation (VI) technique [1, 2, 3] makes it possible to produce functional foods
from fruits and vegetables, keeping their “fresh” physical and sensorial characteristics and
fortifying their nutritional value, by introducing liquids with dissolved or suspended substances
directly into the vegetable porous structure, in a controlled way, allowing fast compositional
changes. Aside from transpiration, respiration is undoubtedly the most important factor
contributing to the deterioration of vegetables after harvest. Information on respiration rates is
needed for the solution of practical problems concerned with storage and transportation of
fresh or minimally processed vegetables. The objectives of this work were to analyze the
effects (i) of VI, (ii) of the presence of aloe vera in VI solutions, and (iii) of temperature, on the
respiration rate and the respiratory quotient of some vegetables.

MATERIALS & METHODS

The vegetables used in the experiments were endive (Cichorium intybus L.), cauliflower
(Brassica oleracea var. Italica), broccoli (Brassica oleracea var. Botrytis L.) and carrot (Daucus
carota L. var. Nantesa).
Vacuum impregnation (VI) experiments were carried out in a specially designed equipment
[2]. The vacuum period of the VI process was performed at 50 mbar during 10 minutes, and
samples remained immersed into the solution of impregnation (SI) during 10 minutes at
atmospheric pressure. Three types of SI were prepared: SucSI (used as VI reference SI), were
aqueous sucrose solutions, isotonic with each of the four raw materials; AV5SI and AV30SI
were aloe vera aqueous dispersions prepared with 5 g/L and 30 g/L of aloe vera powder,
respectively. CO2 production and O2 consumption were determined, and respiratory quotient
calculated, at 5 and 20ºC, for fresh and impregnated vegetables, using a static procedure [5].
Four groups of samples from each vegetable were obtained and their respiration rates
determined and compared: F (fresh, not submitted to VI), VISuc (VI with SucSI), VIAV5 and
VIAV30 (VI with AV5SI and AV30SI, respectively).

RESULTS & DISCUSSION

The amount of VI SI retained by the vegetables, expressed as % of samples initial volume, was
37-46% for broccoli, 7.1-15.0% for cauliflower, 17% for endive, and 10.4-14.7% for carrot.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2089
The three controlled factors (VI, presence of aloe vera, and temperature) had significant effects
on the respiration rate and the respiratory quotient of the studied vegetables.
Respiration rate values were lower at 5ºC than at 20ºC.
For vegetables submitted to VI using the isotonic sucrose solution, and in comparison with
fresh samples, respiration rates at 5ºC were higher for broccoli, endive and carrot, but lower for
cauliflower. At 20ºC, they were higher in the case of broccoli, endive and cauliflower, but
lower for carrot.
As compared with fresh samples, respiration rates at 5ºC were lower for all vegetables
impregnated using the SI with 30 g/L of aloe vera powder. At 20ºC, they were lower for
cauliflower, but higher for broccoli, endive and carrot. The values obtained for the respiratory
quotient were near 1, though the samples impregnated with AV30SI exceeded this value, both
to 5 and to 20ºC.
As an example, Table 1 shows the results obtained in the case of broccoli, the most
impregnated of the four studied vegetables.

Table 1. Respiration data for broccoli

O2 consumption CO2 production
Respiratory Quotient
Treatment mL O2 kg-1 h-1 mL CO2 kg-1 h-1
5ºC 20ºC 5ºC 20ºC 5ºC 20ºC
F 82 ± 20 82 ± 20 71 ± 17 172 ± 51 0.90 ± 0.05 0.99 ± 0.08
VISuc 148 ± 28 148 ± 28 149 ± 30 258 ± 62 1.00 ± 0.02 1.07 ± 0.06
VIAV5 152 ± 18 152 ± 18 153 ± 13 128 ± 15 1.00 ± 0.02 1.0 ± 0.2
VIAV30 106 ± 5 106 ± 5 105 ± 4 318 ± 40 1.00 ± 0.02 1.03 ± 0.06

CONCLUSION
Vacuum impregnation and the presence of aloe vera in vacuum impregnation solutions affect
differently the respiration rate and the respiratory quotient of the different studied vegetables.
This must be taken into account regarding the design of industrial processes of production.

REFERENCES
[1] Fito P., Chiralt A., Betoret N., Gras M.L., Cháfer M., Martínez-Monzó J., Andrés A. & Vidal D.
2001. Vacuum impregnation and osmotic dehydration in matrix engineering: application in functional
fresh food development. Journal of Food Engineering, 49, 175-183.
[2] Salvatori D., Andrés A., Chiralt A. & Fito P. 1998. The response of some properties of fruits to
vacuum impregnation. Journal of Food Process Engineering, 21, 59-73.
[3] Gras M.L., Vidal D., Betoret N., Chiralt A. & Fito P. 2002. The response of some vegetables to
vacuum impregnation. Innovative Food Science & Emerging Technologies, 3, 263-269.
[4] Gras M.L., Vidal D., Betoret N., Chiralt A. & Fito P. 2003. Calcium fortification of vegetables by
vacuum impregnation. Interactions with cellular matrix. Journal of Food Engineering, 56(2-3), 279-
284.
[5] Castelló M. L., Fito P. J. & Chiralt A. 2006. Effect of osmotic dehydration and vacuum impregnation
on respiration rate of cut strawberries. Lebensmittel-Wissenschaft und-Technologie, 39, 1171-1179.

2090
 Production of 4th range Iceberg lettuce enriched with calcium. Evaluation of some

quality parameters

María Luisa Gras a ; Daniel Vidal-Brotóns a ; Fresia Alejandra Vásquez-Forttes a

a
Instituto Universitario de Ingeniería de Alimentos para el Desarrollo (IUIAD), Universidad Politécnica
de Valencia, Valencia, Spain (mgrasro@tal.upv.es)

INTRODUCTION
This project is part of an updating study on the process of production of 4th range Iceberg
lettuce leaves enriched with Ca using the vacuum impregnation (VI) technique. The objectives
of this work were (i) to verify if it is possible that a 250 g portion of impregnated lettuce leaves
provides the same quantity of Ca (300 mg) as a 250 mL glass of milk, which is the 37.5% of
the Recommended Daily Intake (RDI) (800 mg/day), and (ii) to analyze the effects of VI and
of the enrichment with Ca on some quality parameters of the lettuce leaves: water activity (aw),
humidity (water mass fraction, Xw), soluble solids content (Brix), real and apparent densities
(ȡr, ȡa), pH, color (CIEL*a*b* coordinates), mechanical properties (maximum stress and strain,
work performed and number of peaks up to maximum stress), and respiration rate.

MATERIALS & METHODS

Less than 2 days harvested Iceberg lettuces (Lactuca sativa) were used. For the study, three
zones of the whole lettuce leaf were differentiated (on a longitudinal axis), because of the
different distribution of the vascular system: apical (A), medium (M) and basal (B). Vacuum
impregnation (VI) experiments were carried out in a specially designed equipment [1]. The
vacuum period of the VI process was performed at 500 mbar during 10 minutes, and samples
remained immersed into the solution of impregnation (SI) during 10 minutes at atmospheric
pressure. Two SI (SucSI and CaSI) were prepared. SucSI was a sucrose aqueous solution of the
same aw than lettuce leaves, used as a VI reference SI. CaSI (also isotonic with the raw
material) was prepared with 62.5 g of Ca lactogluconate per liter of water, and presented the
following characteristics: Ca content = 5.4 g Ca/L; aw = 0.986±0.003; Brix = 4.13±0.03;
density = 1020±4 kg/m3; pH = 7.3±0.1. Three groups of samples from each zone (A, M, B)
were obtained and their properties compared: F (fresh, not submitted to VI), VISuc (VI with
SucSI), VICa (VI with CaSI). Analytical determinations were performed as described in
previous papers [2, 3; 4].

RESULTS & DISCUSSION

Fresh samples from A, M and B zones showed no differences in aw (0.992) and Xw (0.950-
0.953), but differences in porosity (A: 6.4±0.4%; M: 6.1±0.8%; B: 5.9±0.2%) confirmed the
interest to carry out the study with leaf zone as a controlled factor.
The operation of VI with CaSI led to obtaining Iceberg lettuce with a global content of 169 mg
of Ca in 250 g of impregnated product (200, 156 and 106 mg, for A, M and B zones,
respectively), that is to say the 21% of the RDI. It is probable that the use of more concentrated
solutions of Ca will allow reaching the objective. A fresher, shinier product is obtained without

11th International Congress on Engineering and Food - Athens, Greece, 2011 2091
presenting differences in color. Ca enrichment of the vegetable doesn’t affect significantly its
mechanical behavior, though a light increase of the maximum strength and of the shear work
was detected for B samples, probably due to the interaction of Ca with the wall pectins and
cellular membranes that causes a bigger adhesion between the cells. VI with CaSI doesn’t
modify the respiration rates nor the respiratory quotient of the zone A lettuce samples. For zone
M samples, the intake of O2 increases lightly, without change in the production of CO2,
therefore slightly decreasing the respiratory quotient. For zone B samples, both respiratory
rates increase notably, with an important decrease in the respiratory quotient, indicating that
the reaction seems to prevent the fermentative anaerobic routes.

CONCLUSION
The study shows it seems possible to obtain a Ca enriched product that provides the quantity of
said mineral as a glass of milk, being a possible alternative to dairy products. The industrial
application of this process should include the control of the hydric state of the vegetable
material, with abundant vascular tissue, to increase impregnation capacity and reduce its
variability.

REFERENCES
[1] Salvatori D., Andrés A., Chiralt A. & Fito P. 1998. The response of some properties of fruits to
vacuum impregnation. Journal of Food Process Engineering, 21, 59-73.
[2] Gras M.L., Vidal D., Betoret N., Chiralt A. & Fito P. 2002. The response of some vegetables to
vacuum impregnation. Innovative Food Science & Emerging Technologies, 3, 263-269.
[3] Gras M.L., Vidal D., Betoret N., Chiralt A. & Fito P. 2003. Calcium fortification of vegetables by
vacuum impregnation. Interactions with cellular matrix. Journal of Food Engineering, 56(2-3), 279-
284.
[4] Castelló M. L., Fito P. J. & Chiralt A. 2006. Effect of osmotic dehydration and vacuum impregnation
on respiration rate of cut strawberries. Lebensmittel-Wissenschaft und-Technologie, 39, 1171-1179.

2092
Microencapsulation of Probiotic Bacteria with Alginate and Prebiotic and Evaluation of Survival in
Ice Cream

Cynthia Jurkiewicza, Milena P. M. Boscariolia, Rubia G. Ferreiraa, Eliana P. Ribeiroa, Leo Kunigka

a
Maua Institute of Technology, São Caetano do Sul, Brazil (cynthia@maua.br)

INTRODUCTION

Functional foods are those that promote health benefits beyond basic nutritional functions, when consumed in usual
diet. Prebiotics and probiotics are current examples of functional food ingredients (1). Probiotics are defined as live
microorganisms, which administered in adequate amounts, confer a health benefit to the host. Species of Lactobacillus
and Bifidobacterium, normal components of the intestinal microbiota, are usually employed in many probiotic foods.
Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth
and/or activity of populations of bacteria in the colon. Prebiotics might also enhance the growth and survival of
probiotics in foods. Due to de wide range of therapeutic benefits associated with the consumption of probiotic
microorganisms, there has been an increase in the variety of probiotics foods, including fermented milk, many tips of
cheese, juice, chocolate and also ice cream. Although dairy ice cream seems to be a good vehicle for probiotic cultures
due to its composition and pH near to 6.0, the viability of these microorganisms can be affected by freezing process
and oxygen toxicity. In order to overcome these problems, microencapsulation methods can be applied to increase the
survival of probiotic cultures in frozen dairy products (2).
The aim of this study was to evaluate the influence of prebiotics, resistant starch and acacia gum, incorporated into
alginate beads, on the survival of microencapsulated probiotic bacteria in ice cream over a period of 180 days of
storage at -18 ºC.

MATERIALS & METHODS

Encapsulation procedure
Freeze-dried Lactobacillus acidophilus NCFM and Bifidobacterium lactis BI-04, were provided by Danisco, Brazil.
The prebiotics, acacia gum Fibregum B, was provided by Colloides Naturels, Brazil, and resistant starch, Hi Maize
260, was provided by National Starch, Brazil. Alginate beads were produced using a modified extrusion technique
originally reported by Liserre et al. (3). Solutions of sodium alginate (1% w/v) containing approximately 109 cfu/mL of
Bifidocacterium lactis and Lactobacillus acidophilus, with or without 2% of prebiotic (acacia gum, or resistant
starch,), were atomized in 0.1 M calcium chloride.
Ice cream production and enumeration of probiotic bacteria
Dairy ice cream was formulated with the following composition (w/w): 59% of pasteurized milk, 17% cream, 8.0%
skim milk powder, 12% sucrose, 3.0% glucose and guar gum and carrageenan. The mixture was ripened for 24 h at
5 °C and microencapsulated probiotics were added before freezing in order to achieve approximately 108 cfu.g-1. As
controls, ice cream with free probiotics and microencapsulated probiotics without prebiotics were produced. Each type
of ice-cream was produced in triplicate.
The enumerations of probiotic microorganisms were performed before freezing and at 1, 30, 60, 90,120 and 180 days
of storage at -18°C. Analysis of variance (ANOVA) and Duncan test were applied for determination of significant
difference (p < 0.05) between means of cell counts.
Sensory analyses
Sensory analysis to compare ice cream with free microorganisms (SA) and ice cream with microorganisms
encapsulated in calcium alginate (SB) was performed with 74 untrained panelists using a 9 points hedonic scale (1 =
dislike extremely and 9 = like extremely). The sensory data were analyzed by ANOVA.
Physical and chemical analysis

11th International Congress on Engineering and Food - Athens, Greece, 2011 2093
The mean composition of ice creams (dry matter, ash, fat and protein content) was determined according with standard
methods (4). All measurements were replicated four times. The pH and overrun were also determined.

RESULTS & DISCUSSION

Bifidobacterium lactis counts had no significant reduction (p > 0.05) after freezing and during storage at -18ºC of all
types of ice creams. Encapsulation in calcium alginate with and without prebiotics (acacia gum and resistant starch)
had no effect on survival of B. lactis in ice cream. At the end of 180 days of storage, ice cream with free
microorganism revealed mean counts of B. lactis of 8.19 ± 0.06 log cfu/g, and no significant difference (p > 0.05) were
detected between population of B. lactis in ice cream with free and encapsulated bacteria. With regard to L.
acidophilus population, no significant reduction (p> 0.05) during freezing of ice cream was observed for all types of
ice creams. However, L. acidophilus counts was significantly reduced (p < 0.05), by approximately 0.6 Log cfu/g,
during 180 days of storage at – 18ºC in all types of ice creams. Although significant differences (p < 0.05) were
detected between L. acidophilus counts in ice cream with free and encapsulated microorganisms on days 120, 150 and
180, the difference were less than 0.5 Log cfu/g. To provide health benefits, a minimum of 106 -107 cfu g-1 of probiotic
bacteria must be presented in food in the moment of consumption (FAO). After 180 days of storage, the number of B.
lactis and L. acidophilus in all types of ice creams were above 108 and 107 cfu/g, respectively. This results shows that
the dairy ice cream is an adequate vehicle for probiotic incorporation even without microencapsulation of
microorganisms. Contrary to what was observed in the present study, Homayouni et al. (5) reported that encapsulation
in calcium alginate increased approximately 30% the survival of L. acidophilus and B. lactis in ice cream stored for
180 days at -20ºC. However in the present study the overrun value was 41%, while in Hamayouni et al. (5), the overrun
was 95%. The greater incorporation of air could be responsible for the decrease of viable counts of free
microorganisms in ice cream during storage. The high rate of survival of probiotic microorganisms in this study,
especially free cells, can be justified by the high total solids (33.6%), protein (3.4%) and fat (6.7%), that could protect
or even encapsulate the probiotics. Furthermore, the ice cream pH (6.28), is considered favorable for the survival of
probiotic microorganisms. The sensory score (mean of 74 panelists) of ice cream with free probiotics and ice cream
with encapsulated probiotic was 7.61 and 7.75, respectively. Encapsulation had no significant (p > 0.05) effect on
sensorial acceptability of probiotic ice cream.

CONCLUSION

This study showed that encapsulation of Bifidobacterium lactis and Lactobacillus acidophilus in calcium alginate and
in calcium alginate with acacia gum or resistant starch did not interfere in the survival of microorganisms in ice cream
during 180 days of storage at -18ºC. Moreover encapsulation had no effect on sensorial acceptability of probiotic ice
cream. The numbers of probiotic bacteria in all types of ice cream were above 107 cfu/g at the end of 180 days of
storage. Dairy ice cream can be considered a suitable vehicle for incorporating probiotic microorganisms.

REFERENCES

[1] Siró, I.; Kápolna, E.; Kápolna, B.; Lugasi, A.; 2008. Functional food. Product development, marketing and
consumer acceptance – a review. Appetite, 51(3), 456 – 467.
[2] Anal, K. A.; Singh, H. 2007. Recent advances in microencapsulation of probiotics for industrial applications and
targeted delivery. Trends in Food Science & Technology, 18, 240-251.
[3] Lisere, A.M; Ré, M.I.; Franco, B.D.G.M. 2007. Microencapsulation of Bifidobacterium animalis subsp. lactis in
modified alginate-chitosan beads and evaluation of survival in simulated gastrointestinal conditions. Food
Biotechnology, 21(1), 1-16.
[4] Instituto Adolfo Lutz. Métodos físico-químicos para análise de alimentos. 4. ed. Brasília, DF: Ministério da Saúde,
2005. p.1018.

2094
 The influence of operational parameters in the pectin agglomeration

Talita Akemi Medeiros Hirataa, Vanessa Goulart Machadob, Gustavo César Dacanalc, Florencia Cecília
Menegallia,b

a,b
Department of Food Engineering, University of Campinas, FEA - UNICAMP, 13083-970, Campinas,
São Paulo, Brazil (fcm@fa.unicamp.br)
c
Department of Food Engineering, University of São Paulo, FZEA - USP, 13635-900, Pirassununga, São
Paulo, Brazil. (gdacanal@usp.br)

INTRODUCTION
The agglomeration is usually carried out at equipment where the powder is wetted by a liquid
that promotes adhesion between a particle and the other by linking bridges, leading to
formation of larger particles [1]. Fluid bed agglomeration is commonly used to improve the
instant properties of fine and cohesive food powders. However, the fluidization of fine and
cohesive particles is characterized by cracks and channels. The pulsed fluid bed has some
advantages over the conventional fluid bed equipment, including easy fluidization of irregular
particles of different sizes [2, 3, 4]. Pectin is widely used in the food industries as a gelling
agent and has been used in powder form. Thus, the aim of this work was to study the influence
of some operational parameters in the agglomeration process of pectin and to determine the
optimal process conditions which lead to a higher size increase.

MATERIALS & METHODS

The product used in this study was commercial high ester pectin, extracted from citrus peel,
(brand Genu type 105 rapid set CP Kelco). The runs were performed in a pulsed fluidized bed.
The experimental runs were done according to a full factorial design 24. The independent
variables at the following interval levels were: fluidizing air temperature (60 to 90 °C),
fluidizing air velocity (0.36 to 0.68 m/s), binder flow rate (0.0 to 1.6 ml/min) and air pulsation
frequency (0 to 800 rpm). The dependent variables or experimental responses were: process
yield (Yld), calculated as the ratio between the mass of samples of raw and agglomerated
product; product moisture (Mst), powder samples were dried under vacuum at 70 ° C until the
weight becomes constant; flowing out fluidizing air relative humidity (rhout), obtained from wet
and dry bulb temperatures measurement; and mean particle diameter (dpm) were determined by
using the equivalent diameter, calculated by the projected area of the particles. The minimum
fluidizing air velocity was determined visually observing the beginning of movement of the
bed.

RESULTS & DISCUSSION

The results of the experimental design were statistically analyzed using the software
STATISTICA v.7.0 (StatSoft, Inc., USA). The results for mean particle diameter showed that
the binder flow rate is the main independent variable that influences the results. When binder
flow rate was higher the particles surface become most sticky, then the adhesion mechanisms
were enhanced. Lower temperatures of drying that preserves higher moisture content of
particle surfaces and higher binder flow provide higher adhesion among a greater number of

11th International Congress on Engineering and Food - Athens, Greece, 2011 2095
particles, increasing the diameter of the cluster formed at these conditions. The statistical
analysis for air pulsation frequency did not show significant effects on responses, when
compared with others independent variables. In relation to the raw material, the mean particle
diameter increased from 59.28 μm to 259.15 μm. The product moisture remained low at 7.96%
db, but higher than the moisture content of the raw material (6.7% db.). Process yield was
above 80% and the flowing out fluidizing air relative humidity remained low (51.2%). The
distribution of particle size can be observed in Figure 1.

Figure 1. Experimental and simulated time temperature profile.

Figure 1. Particle size distribution for raw material and product cluster obtained at optimum process.

CONCLUSION
It can be concluded that the variable most important was the flow of binder, since the higher
values provokes higher process yields (81.85%), increase in particle size. Moreover, the
product loss for incrustation on the wall of the bed and lump formation at conditions tested can
be considered negligible due to the moisture content of the particles remain at a low value.

REFERENCES
[1] Ax, K., Feise, H., Sochon, R., Hounslow, M., Salman, A. 2008. Influence of liquid binder dispersion
on agglomeration in an intensive mixer. Powder Technology, 179, 190-194.
[2] Dacanal, G.C. 2009. Agglomeration of acerola powder and soy protein isolate in a conical pulsed-
fluid bed. PhD Thesis, College of Food Engineering, University of Campinas, UNICAMP, Brazil,
202p.
[3] Gawrzynski, Z.; Glaser, R.; Kudra, T. 1999. Drying of powdery materials in a pulsed fluid bed dryer.
Drying Technology, 17 (7-8), 1523–1532.
[4] Reyes, A., N. Herrera and R. Vega 2008, Drying suspensions in a pulsed fluidized bed of inert
particles. Drying Technology, 26, 122-131.

2096
 Antioxidant activity of microcapsules of Rubus sp. juice using spray drying

Jimenez M.a, Azuara E.a, Vernon-Carter J.b, Luna-Solano G.c, Beristain C.I.a

a
Instituto de Ciencias Básicas, Universidad Veracruzana, Xalapa Ver, México (maribjimenez@uv.mx)
b
Universidad Autónoma de México, Distrito Federal, México
c
DEPI, Instituto Tecnológico de Orizaba, Orizaba, Veracruz, México

INTRODUCTION
Blackberries (Rubus fruticosus) contain large amounts of anthocyanins, and these flavonoid
pigments give blackberries their characteristic red to blue color [1]. There are many factors
which affect the stability and intensity of colorants during processing and storage. It is reported
that its stability is affected during processing and storage [2]; therefore encapsulation by spray
drying was used to avoid the colorant degradation. The storage stability of powdered
antioxidants for extended periods is of interest to manufacturers and consumers of food
packaged for long-term storage. In view of the foregoing, the objective of the present work was
to develop and characterize a powder by spray drying that is derived from Rubus fruticosus
juice and to evaluate its antioxidant stability.

MATERIALS & METHODS

The fruits were purchased at three of the local markets during the winter and transferred to the
University. The total solid content and pH was determined according to AOAC [3]. Juice was
added at a 1:3 ratio (w/w) with respect to the GA contained in the solution. The mixture was
homogenized and then it was dried in a Mini Spray Dryer model Büchi 190 with inlet and
outlet air temperatures of 160 + 5 °C and 90 + 5 °C, respectively. The anthocyanins content
was determined according to Elisia et al. [4]. The L, a, b color values of the samples were
measured using a spectral photometer (Color Flex CX1115 HunterLab USA). The moisture
content of the powder was determined gravimetrically by oven-drying at 60 º C until constant
weight was attained [3]. The antioxidant properties were determined by DPPH radical
scavenging activity, redox potential, reducing power and optical density [5-7]. The samples
were examined by a JSM-5600LV scanning electron microscope.

RESULTS & DISCUSSION

The characteristics from Rubus juice are shown in Table 1. In the present work, the pH for the
natural juice of fruit was 3.8 and 4.5 for the capsules. The soluble solids in the natural juice
were 15 %. The anthocyanin content was similar to that reported for others authors (420 mg/L).
The antioxidant activity express by DPPH radical activity was 75%. The results obtained
showed that there was a diminution of 74% in the antioxidant activity when samples were
encapsulated. In regard to the concentration of antocianins a decrement of 66% was observed
in the concentration in microcapsules (280 mg/L) compared with the natural juice of Rubus sp.
On the other hand, the redox potential in juice (480 mV) was similar to microcapsules (450
mV), thus indicating the probable formation of compounds able to promote the transference of
electrons. Reducing power was similar in juice and microcapsules, indicating a good
antioxidant activity for microcapsules obtained.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2097
Table 1 Initial values of analysis performed on natural Rubus sp. juice and capsules obtained by aspersion

Determination Juice Capsules

Total solid content 15 30

Moisture content (%) 85 11
Water activity 0.997 0.576
% antioxidative activity (DPPH) 75 56
pH 3.80 4.50
Reducing power 0.82 0.987
Anthocyanin concentration (mg/L) 420 280
Redox Potential (mV) 480 450
Polyphenol content (mg/100 g) 850 1230
Color L*=7.92 L*=3.78
a* = 16 a* = 8.50
b* = 4.9 b* = 2.35

The microcapsules were equilibrated at water activity of 0.576 and exhibited a very good
homogeneous morphology: spherical shaped, smooth surfaced particles that varied greatly in
size and were free of visible cracks and pores. The presence of surface dents may be due to
uneven shrinkage during drying.

CONCLUSION
The present study indicates that the fruit from Rubus sp. is rich source of anthocyanin, phenols
and antioxidants, demonstrating its potential use as a food additive. Spray drying and the used
of Gum Arabic as wall material providing an effective protection to antioxidant compounds
present in Rubus sp. juice. Therefore, the microcapsules described in this study represent an
interesting food additive for incorporation into functional foods, both as an antioxidant and as a
colorant.

REFERENCES
[1] Koca I. & Karadeniz B. 2009. Antioxidant properties of blackberry fruit grown in the black sea
region of turkey. Scientia Horticulturae, 121, 447-450.
[2] Attoe E.L. & von Elbe J.H. 1982. Degradation kinetics of betanine in solutions as influenced by
oxygen. Journal of Agricultural and Food Chemistry, 30, 708-712.
[3] AOAC. 1995. Official Methods of Analysis, CUNNIF, P. (Ed). Association of Official
Analytical Chemists International, Arlington, Virginia, USA.
[4] Elisia I., Hu C., Popovich D. & Kitts D. 2007. Antioxidant assessment of an anthocynin-
enriched blackberry extract. Food Chemistry, 101, 1052-1058.
[5] Kitts D.D., Wijewickreme A.N. & Hu C. 2000. Antioxidant properties of a North American
ginseng extract. Molecular and Cellular Biochemistry 203, 1–10.
[6] Manzocco L., Anese M. & Nicoli C. 1998. Antioxidant properties of tea extracts as affected by
processing. Lebensmittel Wissenchaft und Technologie, 31, 694-698.
[7] Oyaizu M. 1986. Studies on products of browning reaction prepared from glucose amine.
Japanese Journal of Nutrition, 44, 307-315.

2098
 Novel ways to control enzymatic hydrolysis as a tool to produce functional peptides

Elena Leeb a, Ulrich Kulozikb, Seronei Cheison a

a
Technische Universität München, Junior Research Group: Bioactive Peptides and Protein Technology,
Freising Weihenstephan, Germany (Elena.Leeb@wzw.tum.de)
b
Technische Universität München, Chair for Food Process Engineering and Dairy Technology, Freising
Weihenstephan, Germany

INTRODUCTION
To date, processes to obtain and enrich functional peptides in several foods are not feasible, though
different milk proteins are known to be potential sources for functional peptides. Particularly ȕ-
Lactoglobulin (ȕ-Lg) is a source for several peptides with different biofunctionalities like
antibacterial and hypocholesterolemic activity as well as angiotensin-I converting enzyme (ACE)-
inhibitory properties . Currently, the release of peptides by means of enzymatic hydrolysis results in
a mixture of numerous different peptides. A new approach to control enzymatic hydrolysis to
increase the release of desired peptides is the pre-treatment of the substrate. Thereby conformational
changes within the protein structure prevent the hydrolysis of undesired, and increase the
accessibility of the enzyme to desired peptide bonds. The influence of conformational changes
induced by high hydrostatic pressure [1] and thermal treatment [2] on the hydrolysis performance
has already been investigated. However, to date there is still no specific information about a thermal
pre-treatment of the substrate on the released peptides during hydrolysis.
Therefore the aim of this study was to determine the influence of thermal denaturation of ȕ-Lg on
the release of functional peptides by subsequent controlled enzymatic hydrolysis.

MATERIALS & METHODS

Bovine ȕ-Lg with a purity of 95.7 % was prepared out of whey protein isolate as described by
Gezan-Guizou et al. [3] using an optimised method. Aqueous ȕ-Lg solutions (50 g/L) were adjusted
to pH 8 and completely denatured by thermal treatment at 80°C. The native as well as the denatured
ȕ-Lg was hydrolysed with trypsin (T9201 from bovine pancreas, activity of t 7500 units/mg solid,
Sigma-Aldrich) at the enzyme optimal conditions (38°C, pH 8) with an Enzyme-to-Substrate-Ratio
= 0,1 %. During the hydrolysis experiments the pH was kept constant by addition of 0.5 M NaOH
and thus, the Degree of Hydrolysis (DH) could be calculated, based on the consumption of Base [4].
The peptide composition of the sampled aliquots at DH 1 %, 5 % and maximum possible DH
(DHmax) were analysed by matrix-assisted laser desorption/ionisation time-of-flight tandem mass
spectrometry (MALDI-TOF-MS/MS) as already described in our earlier work [5].

RESULTS & DISCUSSION

It was shown that thermal denaturation of ȕ-Lg at pH 8 results in the formation of the so-called non-
native monomers. At subsequent tryptic hydrolysis of the native and the non-native ȕ-Lg monomers,
differences in the released peptides were determined. The MS analysis exhibited that denaturation
decreased the susceptibility of Trypsin to possible cleavage sites during the hydrolysis. Looking at
the order of cleaved peptide bonds within the native protein during tryptic hydrolysis, no
exceptional specificity towards individual peptide bonds could be estimated. In contrast,
denaturation led to the restricted cleavage of the peptide bonds Arg40, Lys69, Lys70, Lys141 and Arg148

11th International Congress on Engineering and Food - Athens, Greece, 2011 2099
at DH 1%. This limitation can be explained by the unfolding of the monomer and the increased
exposure of these regions. Due to these thermal induced structural changes the peptide f(142-148)
with known ACE inhibitory activity was released already at the outset of hydrolysis.

Table 1. Identified cleaved peptide bonds within the native and non-native monomer of ȕ-Lg
Identified cleaved peptide bonds within the
Trypsin specific
native ȕ-Lg monomer non-native ȕ-Lg monomer
cleavage sites
DH 1 % DH 5 % DHmax DH 1 % DH 5 % DHmax
Lys8 Ɣ Ɣ Ɣ
Lys14 Ɣ Ɣ Ɣ ż ż
Arg40 Ɣ Ɣ Ɣ ż ż ż
Lys47 Ɣ Ɣ ż
Lys60 Ɣ Ɣ Ɣ ż ż
Lys69 Ɣ Ɣ Ɣ ż ż ż
Lys70 Ɣ Ɣ Ɣ ż ż ż
Lys75 Ɣ Ɣ
Lys77 Ɣ Ɣ Ɣ ż
Lys83 Ɣ Ɣ Ɣ ż
Lys91 Ɣ Ɣ Ɣ ż ż
Lys100 Ɣ Ɣ ż
101
Lys Ɣ Ɣ Ɣ
Arg124 Ɣ Ɣ ż ż
Lys135 Ɣ Ɣ Ɣ ż
Lys138 Ɣ
Lys141 Ɣ Ɣ Ɣ ż ż ż
Arg148 Ɣ Ɣ Ɣ ż ż ż

CONCLUSION
The results of this study demonstrate that a thermal pre-treatment of ȕ-Lg is an effective way to
control and steer enzymatic hydrolysis. By appropriate choice of denaturation conditions the
accessibility of the enzyme to individual peptide bonds can be controlled and thereby the release of
desired peptides promoted.

REFERENCES
[1] J.C. Knudsen, J. Otte, K. Olsen, L.H. Skibsted. 2002. Effect of high hydrostatic pressure on the conformation of
[beta]-lactoglobulin A as assessed by proteolytic peptide profiling. International Dairy Journal, 10, 791–803. [2] N.
Stanciuc, A. Hintoiu, S. Stanciu, G. Rapeanu. 2010. Thermal treatment can modify the susceptibility of whey protein
concentrate to enzymatic hydrolysis. Innovative Romanian Food Biotechnology, Vol. 7, Issue of September, 30–36. [3]
G. Gesan-Guiziou, G. Daufin, M. Timmer, D. Allersma, C. van der Horst. 1999. Process steps for the preparation of
purified fractions of -lactoglobulin from whey protein concentrates. Journal of Dairy Research, 02, 225–236. [4] J.
Adler-Nissen. Enzymic hydrolysis of food proteins, Elsevier Applied Science Publishers; Sole distributor in the USA
and Canada; Elsevier Science Pub. Co., London ;, New York, New York, NY, USA. 1986. [5] S.C. Cheison, M.-Y.
Lai, E. Leeb, U. Kulozik. 2011. Hydrolysis of [beta]-lactoglobulin by trypsin under acidic pH and analysis of the
hydrolysates with MALDI-TOF-MS/MS. Food Chemistry, 4, 1241–1248.

2100
 Influence of the structure and composition of the País grape proanthocyanidins on the

inhibition of angiotensin converting enzyme

Susana Godoy, Marlene Roeckel, Estrella Aspé, Katherina Fernández

Chemical Engineering Department, University of Concepción, Chile (kfernandeze@udec.cl)

INTRODUCTION
Previous works developed in our laboratory showed that the País grapes from the Itata Valley,
Chile, have high amounts of Proanthocyanidins (PAs) when compared with another varieties,
like Carmenere or Pinot Noir [1]. The PAs are located mainly in the skin and seeds of the
grapes and they are phenolic compounds of high complexity, with molecular weight over 500.
They are part of the flavonols family and are polymers composed by flavan-3-ol subunits
connected by C-C bonds. Their sizes get higher as the grape gets mature, and it has being
reported that their bioactive properties are determinate by molecular composition and size [2].
A bioactivity reported for the PAs had been the inhibitory effect on Angiotensin-Converting
Enzyme, ACE [3]. The understanding of the influence of PAs composition and size on ACE
inhibition by PAs is still not very clear. This information is highly relevant when trying to
determinate the inhibition mechanism and to enhance its use and to develop a natural medicine.
The main objective of this work was to evaluate the influence of the number of subunits (mDP)
and the type of subunits (C, EC, ECG, EGC) of the PAs extracted from the seed and skin of
País grapes over ACE activity, for which the PAs of different molecular sizes present in the
seed and skin extracts were separated, then the composition and size of each group was
determinate, after which their inhibitory activity over ACE was confirmed and determinate, to
finally find dependences between the fractions properties and their inhibitory ability.

MATERIALS & METHODS

The grapes were obtained from Quillón Valley, Bio Bio Region, Chile. The PAs extraction was
based on Villarroel [1]. Both extracts were separately injected into a gel permeation
chromatography column packed with Toyopearl HW-40F resin, following the protocol
described by Kennedy and Taylor [4]. Five fractions from skin and 5 fractions from seeds were
isolated, which were concentrated in rotary evaporator, and lyophilized to a dry powder for
further analysis (Labconco, Freezer Dry System, USA). The composition of each fraction was
study by acid-catalysis in the presence of excess Phloroglucinol. The PAs inhibition on ACE
(Sigma-Aldrich, rabbit-lung ACE; A6778, USA) was carried out, testing crude extracts, each
purified fraction and Enalapril Maleato (Laboratorios Chile, Enalapril Maleato 10mg, Chile) as
inhibitors.

RESULTS & DISCUSSION

The PAs inhibitory capacity over ACE was studied by determination of the IC50 (μM) by data
extrapolation. Then, the activity of each fraction was related to their mDP and structural
characteristics. For the skin fractions (Table 1), it can be observed that FP-A and FP-B were

11th International Congress on Engineering and Food - Athens, Greece, 2011 2101
the less effective inhibitors, while the higher polymers had an increasing of the inhibitory
capacity. In relation to composition, it can be noticed an inverse correlation between the
amount of C and the inhibitory capacity of the fractions and a strong predominance of the EC
and EGC subunits in the fractions with higher mDP, which could contribute to its better
inhibitory ability, since this last mentioned monomer has being reported form strong
interactions with proteins because of its 3 hydroxyl groups.

Table 1. IC50 of each fraction related to their subunits compositions and mDP for skin fractions.
Extension subunits
Termina subunits
IC50 (phloroglucinol adducts)
mDP
(μM)
EGC C EC ECG C EC ECG

FP-A 4.108 2.49 8.0 6.6 45.0 0.0 40.32 0.0 0.0
FP-B 7.208 2.7 10.3 5.7 47.2 0.0 36.67 0.0 0.0
FP-C 0.014 9.4 14.9 3.5 67.6 10.7 3.43 0.0 0.0
FP-D 0.048 16.14 26.0 1.4 66.3 0.0 6.20 0.0 0.0
FP-E 0.0164 58.85 30.8 1.6 65.9 0.0 1.70 0.0 0.0

When comparing seed and skin fraction, fractions FP-B and FS-B with similar mDP, the main
difference was the amount of EC, being higher in the last one. Also, this sample had the better
inhibitory capacity, which can indicate a relation between both factors. For the chemical
inhibitor Enalapril, a commonly used antihypertensive medicine, on ACE inhibition, the results
show an EC50=0.107 μM value of inhibitory capacity higher than the small fractions (FP-A,
FP-B and FS-A), similar to the medium-sized fractions (FS-B and FS-C) and worst than the big
fractions (FP-C, FP-D, FP-E, FS-D and FS-E).

CONCLUSION
All the fractions obtained from skin and seed extract of País grapes had a verified inhibitory
activity over ACE, being the FP-C the most effective one.

REFERENCES
[1] Villarroel M. 2009. Caracterización del contenido de Proantocianidinas en las uvas País Negra (Vitis
vinífera L) del Valle del Itata. Memoria de Titulo en Departamento de Ingeniería Química,
Universidad de Concepción: Concepción. p. 63.
[2] Bindon K.A., Smith P.A., Holt H., Kennedy J.A. 2010. Interaction between Grape-Derived
Proanthocyanidins and Cell Wall Material. 2. Implications for Vinification. Journal of Agricultural
and Food Chemistry, 58(19), 10736-10746.
[3] Actis-Goretta L., Ottaviani J.I. & Fraga C.G. 2006. Inhibition of angiotensin converting enzyme
activity by flavanol-rich foods. Journal of Agricultural and Food Chemistry, 54(1), 229-234.
[4] Kennedy J.A. & Taylor A.W. 2003. Analysis of proanthocyanidins by high-performance gel
permeation chromatography. J Chromatogr A, 995(1-2), 99-107.

2102
 Kinetic characterization of inhibition of angiotensin converting enzyme by

proanthocyanidins extracted from vitis vinífera L. cv. País

Karen Álvarez, Marlene Roeckel, Estrella Aspé, Katherina Fernández

Chemical Engineering Department, University of Concepción, Concepción, Chile (kfernandeze@udec.cl)

INTRODUCTION
Hypertension is considered one of the most important and common global public health,
associated to high mortality affecting mainly the developed countries. Its treatment usually
involves pharmaceutical drugs to control the patient's systemic change, where specific
inhibitors of angiotensin-converting enzyme (ACE) are used [1]. The existence of ACE
inhibitors from natural foods, rich in phenolic compounds, have been demonstrated,
specifically flavan-3-ols [2]. The flavan-3-ols polymerized to form the proanthocyanidins
(PAs), which are chains of monomers catechin, epicatechin and gallic acid esters [3]. The main
objective of this study was to characterize the ACE inhibition type using extracts rich in PAs
from grape Vitis vinifera L. País obtained from the skin and seed.

MATERIALS & METHODS

The skin and seeds of 200 grapes were manually separated and extracted separately in
Erlenmeyer flasks with 250 mL of acetone: water (2:1 v/v) for 15 h with continuous stirring at
35 °C and in the absence of light to reduce oxidation. The raw extracts were purified separately
by size exclusion chromatography using as packing Toyopearl HW-40F resin. The kinetics
assays of ACE inhibition by PAs purified extracts from skin and grape seed, were performed as
follows: a 50 mM buffer Tris-HCl was used with 300 mM NaCl and pH 8.3 to prepare a
reaction volume consisting of an aliquot of 60PL of HHL (0.5-5 mM) and 10PL of ACE mixed
with skin or seed extract (60PL, 0-1 mg/mL), pre-incubated separately for 10 minutes at 37 °C.
Reaction was carried out in polyethylene tubes of 1.5 mL, the HHL was added to the ACE
mixture and kept for 80 minutes at 37 °C. The reaction was stopped with 170 P L of HCl (1 M)
and the samples filtered with PTFE disposable filters (0.45 Pm). The analysis was performed
using a previously described HPLC method [4]. The data were adjusted to linearization of
Michaelis Menten kinetic and they were evaluated by equation Lineweaver-Burk, Eadie-
Hofstee and Lagmuir.

RESULTS & DISCUSSION

The experimental data for seed and skin extracts were adjusted to Michaelis Menten kinetic
model, Figure 1A y Figure 1B, respectively. The plots showed a dose-dependent behaviour in
which the HA production increased with increasing of substrate concentration. Also, for both
extracts when increasing the PA concentration a higher ACE inhibition was observed, since the
curves were positioned under the control curve (assay performed without inhibitor). It can be
seen that the skin extract (B) inhibited more than the seed (A), showing initial rates under 1.4
(uM HA/min) versus 1.7 (uM HA/min), respectively.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2103
 A B

Figure 1. Data adjusted to Michaelis Menten kinetic on ACE inhibition kinetics by (A) seed and (B) skin
extracts. Inhibitor concentrations of (+) control, (i) 0.001 mg/mL, (Ÿ) 0.005 mg/mL, (×)0.01 mg/mL,
(ŷ) 0.1mg/mL and (Ɣ) 1mg/mL

Linear adjustments of the three models predicted a mixed inhibition for both extracts, since a
variation considerable in the maximum enzyme rate (vmaxapp), and in the Michaellis-Menten
constant (Kmapp) was observed. Thus, in the range of concentration studied, the inhibitor (PAs)
affected both kinetic parameters.

CONCLUSION
This study showed a mixed kinetic inhibition for seed and skin extracts on ACE inhibition. The
inhibition constants (Km y vmax) were in the same range and comparable to previous studies.
These results might indicate that PAs from this grape variety allows an adequate interaction
with ACE due to the PAs chain size and conformation. It was established that skin inhibits
much better than the seed for their structural differences and availability of OH groups.

REFERENCES
[1] Chobanian A.V., Bakris G.L., Black H.R., et al. 2003. Seventh Report of the Joint National
Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure.
Hypertension, 42, 1206-1252.
[2] Actis-Goretta L., Ottaviani J.I., Keen C.L. & Fraga C.G. 2003. Inhibition of angiotensin converting
enzyme (ACE) activity by flavan-3-ols and procyanidins. Febs Letters, 555, 597-600.
[3] Schofield P., Mbugua D.M., & Pell A.N. 2001. Analysis of condensed tannins: a review. Animal
Feed Science and Technology, 91, 21-40.
[4] Eriz G., Sanhueza V., Roeckel M., & Fernández K. 2011. Inhibition of the angiotensin-converting
enzyme by grape seed and skin proanthocyanidins extracted from Vitis vinífera L. cv. País. LWT-
Food Science and Technology. LWT - Food Science and Technology 44, 860-865.

2104
 Enzymatic depolymerisation of oat ȕ-glucan

Aliki-Ilona Niniosa, Juhani Sibakovb, Ioanna Mandalaa, Konstantinos Fasseasa, Kaisa Poutanenb, Emilia
Nordlundb, Pekka Lehtinenb

a
Department of Food Science and Technology, Agricultural University of Athens, 75 Iera Odos, 11855,
Athens, Greece (alikininiou@gmail.com, imandala@aua.gr)
b
VTT Technical Research Centre of Finland, P.O.Box 1000, Tietotie 2, Espoo, FI-00244, VTT
(juhani.sibakov@vtt.fi)

INTRODUCTION
The aim of the present study was to modify the molecular weight and viscosity properties of ȕ-
glucan in oat bran by hydrolytic enzymatic treatment. Water content, enzyme type and dosage
and reaction time were used as variables in studying and optimizing the reaction conditions for
ȕ-glucan hydrolysis. Oat bran fractions with 20.0 and 28.5 % of ȕ-glucan were treated with
commercial enzyme mixtures and with purified Trichoderma reesei endo-glucanase II. After
the hydrolysis reaction, ȕ-glucan was extracted with hot water at 80 ºC. The solids were
removed by centrifugation, and the supernatant stored at 5 ºC. Rheological properties and
molecular weight distribution of the enzyme-treated ȕ-glucan, as well as the stability of the
solution were then measured. The tested reaction parameters significantly affected the
hydrolysis of ȕ-glucan, suggesting that careful optimization of the reaction conditions is
needed when aiming at specific hydrolysis products.

MATERIALS & METHODS

Two oat bran concentrates were used as raw materials. They were manufactured either from
non-heat-treated whole oat kernels (OBC-1) or from commercial heat-treated oat bran (OBC-2)
according to Sibakov et al. [1]. The ȕ-glucan concentrations of OBC-1 and OBC-2 were 28.5
% and 20.0 (dw), respectively. The oat bran concentrates were enzymatically hydrolysed either
at 50 % (low) or at 94 % (high) water content. The process in low water content was based on
the efficient mixing of OBC and enzyme, and subsequent incubation of the dough-like mass at
50 ºC for 10 min – 4 h. The hydrolysis at higher water content was performed by using a
continuous mixing of the OBC-enzyme suspension at 45 ºC for 1 or 4 h (Figure 1). After both
processes, the enzyme was inactivated by high temperature (115 or 100 ºC) treatment. Both a
commercial enzyme Depol 740 L (Biocatalyst Ltd, Wales, UK) and a purified Trichoderma
reesei endo-glucanase II (EGII, VTT, Finland) were investigated.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2105
 High water content

Oat bran concen trate (O BC) B Suspen sion of oat

Water
Low water content bran concentrate

A Enzyme
Water

Preconditioning at 40 ºC Mixing of the solution

Water in flasks at 45 ºC
for 30min, 25 % moisture
Extraction for 30–120 min

Hydrolysis in extruder at
Enzyme Separation of solids
45 ºC for 1–2 min, Separation of
and water
50 % moisture solid s

InactivatIon
Incubation at 50ºC for Stabilisation & of enzymes at boiling
10 min–4 h, 50% moisture Pasteurisation water for 10 min

Inactivation in extruder Low viscosity Low visco sity

at 115ºC for 1–2 min ß-glu can solution ß-glucan solution

Figure 1. The flow chart of the enzymatic depolymerisation of ȕ-glucan by extrusion process at low (A)
and high water content (B).
RESULTS & DISCUSSION
Enzymatic hydrolysis at high water content
A 1.0 % solution of oat bran concentrate OBC-1 was hydrolysed by different commercial
enzyme mixtures and by the purified T. reesei EG II. After the hydrolysis at 94 % water
content, the concentrations of over 10 kDa ȕ-glucans were reduced by as much as 90 %.
According to the results, the time of enzymatic treatment is essential, as a long time (4 h)
resulted in extensive ȕ-glucan degradation.
Enzymatic hydrolysis at low water content
By using the lower water content, lower degradation of ȕ-glucan and higher average molecular
weights compared to the high water content were obtained. For example, after 4 h hydrolysis
with Depol 740 L preparation, the molecular weight at low water content was about 2-fold
greater compared to the hydrolysis at high water content.

CONCLUSION
This study showed an efficient way to tailor the molecular weight of oat bran ȕ-glucan by
enzymatic hydrolysis at limited water content. The suitable range of ȕ-glucan molecular weight
was obtained when the oat bran concentrate was treated at 50 % moisture content at 50 ºC for
more than 3 hours by hydrolytic Depol 740 L enzyme preparation. The peak average molecular
weight of ȕ-glucan after such hydrolysis was 47 kDa. The highest concentration of ȕ-glucan
which did not convert into gel during the 18 days storage at 5 ºC was 2.0 %. In spite of the
uncertainty of LMW ȕ-glucan’s cholesterol lowering affect, the results could be used as a base
for development of dietary fibre-enriched liquid products.

REFERENCES
[1] Sibakov, J., Myllymäki, O., Hietaniemi, V., Pihlava, J.-M., Kaukovirta-Norja, A., Poutanen, K.,
Lehtinen, P., 2010, Combination of defatting and dry fractionation technologies to produce oat
ingredients with high ȕ-glucan concentration In: Dietary Fibre: New frontiers for food and health,
van der Kamp, J., McCleary, B. & Topping, D. (Eds.), Wageningen Academic Publishers, The
Netherlands, pp. 79–89.

2106
 Parameters evaluation of fructooligosaccharides production by sucrose
biotransformation using an osmophilic Aureobasium pullulans strain

Juliana Bueno da Silvaa, Ana Elizabeth Cavalcante Faia, Rosângela dos Santosa, Luis Carlos
Bassob, Gláucia Maria Pastorea
a
University of Campinas, College of Food Engineering, Campinas, Brazil
b
University of São Paulo, ESALQ, Piracicaba, Brazil (jubueno@fea.unicamp.br)
INTRODUCTION
The fructooligosaccharides (FOS) belonging to the prebiotics group that are “non-digestible
oligosaccharides food ingredients but fermentable by the bacteria in the gut microbiota. They
selectively promote growth of the beneficial bacteria (lactobacilli and bifidobacteria) and
provide a series of benefits to the human health. Such effects include activation of the human
immune system, maintenance the intestinal microbiota, resistance to infection, enhanced
mineral absorption by the gastrointestinal tract, synthesis of B complex vitamin, lowering of
serum cholesterol and preventing carcinogenic tumors [1]. Industrial production of this
ingredient was mainly done using the enzyme fructosyltransferase of Aspergillus niger and
Aureobasidium sp. reaching yield value around 60% and 53-59% respectively [2]. In our study,
the synthesis of FOS was performed from sucrose metabolized by the role cells of
Aureobasidium pullulans, isolated from honeycomb through RSM (Response Surface
Metodology) using a Plackett-Burman matrix with 16 assays (PB-16) to evaluate 12 variables:
concentration of sucrose, inoculum, yeast extract, urea, K2HPO4, (NH4)2SO4, MgSO4, ZnSO4,
MnSO4. This method is a screening approach used to statistically select the signi¿cant
variables of numerous factor-experiments [3].
MATERIALS & METHODS
The culture of Aureobasidium pullulans was maintained in YEPD slants, containing glucose
2% (w/v), yeast extract 1% (w/v), peptone 1% (w/v), agar 1.5 % (w/v) at 4°C. For the pre-
inoculum cultive a loopful of cells was streaked into YEPD plate, for 48h at 30°C. This was
transferred to 150 mL culture medium of cane molasses containing 6% (w/v) of total reduce
sugars for 48h, 150 rpm at 30°C. The suspension was centrifuged at 10.000 rpm for 15 minutes
and the cells were used for initial inoculum for the experiments that were done according
strategy Plackett-Burman . The levels of each variable (%) were sucrose: 20 a 40; yeast extract:
0 a 0.5; inoculum: 1 a 20; K2HPO4: 0 – 0.0435; urea: 0 – 0.015; (NH4)2SO4: 0 – 0.033;
MgSO4.7H2O: 0 – 0.0245; ZnSO4: 0.0015; MnSO4.7H2O: 0 – 0.001, pH: 4.5 a 6.0; temperature
(°C): 27 a 30; agitation (rpm): 150 a 250. Fermentations proceeds in 125 mL flasks with 20 mL
of medium and the sampling were done at 0, 24, 48 and 72 hours. The sugar mesuring was
dones by HPLC, column Lichrospher 100 NH2, 26°C, 1 mL/min with acetonitrile:water 70:30
(v:v) as mobile phase.
RESULTS & DISCUSSION
The effects of the above 12 variables on yield, which were calculated through formula: Y=
[total FOS]*100/ [initial sucrose] and concentration of total FOS % (g/100 mL were shown in
Table 1. The statistical analysis were conducted by the software STATISTICA® 7.0
considering a significant level of 10% (P<0.1).

11th International Congress on Engineering and Food - Athens, Greece, 2011 2107
 Table 1. Experiments results in Plackett-Burman
0 hour 24 hour 48 hour 72 hour
Treat-ment (T)
FOS % Yield FOS % Yield FOS % Yield FOS % Yield
1 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
2 6.58 16.45 7.71 18.03 8.49 18.35 3.60 7.32
3 11.81 29.52 15.03 32.32 8.63 16.27 8.42 16.51
4 0.00 0.00 20.52 49.97 10.22 21.78 11.2 23.94
5 6.91 34.57 2.98 14.91 0.53 2.63 0.00 0.00
6 0.00 0.00 7.05 14.45 3.55 8.86 3.63 6.65
7 4.86 24.31 0.00 0.00 0.00 0.00 0.00 0.00
8 7.68 19.20 12.67 31.78 9.77 24.44 5.46 13.64
9 4.00 10.00 9.57 22.60 22.30 54.70 4.22 9.05
10 6.70 33.49 1.63 8.15 1.21 6.09 1.27 6.36
11 1.35 6.76 3.87 19.33 2.82 14.09 0.72 3.61
12 1.42 3.55 20.34 49.34 16.96 38.16 14.5 30.46
13 0,00 0,00 1,00 5,00 0,00 0,00 0,00 0,00
14 0,03 0,14 9,60 45,94 5,94 29,49 3,49 13,76
15 0,15 0,74 7,05 35,26 3,13 15,64 3,70 18,51
16 1,05 5,24 6,14 30,73 5,88 29,40 6,51 32,53
PC1 3,62 12,07 8,19 27,29 3,76 12,52 1,61 5,37
PC2 3,62 12,07 8,40 27,96 3,48 11,62 3,69 12,30
PC3 3,62 12,07 9,23 30,76 3,60 12,01 3,34 11,15
The results showed that high yields reach values of 54.7% at 48h of reaction in T9, 49.97% at
24h in T4 and 49.34% in 24h in T12. For the FOS Yield data showed significance at 24h the
variables agitation, with negative effect -16.32 and p-value 0.016; MnSO4, with positive effect
15.33 and p-value 0.020. At 48h the variable was significant only agitation with negative effect
-14.94. The inoculum level showed significance at 0h with positive effect 14.08, however it
was negative at 24 and 48. Regarding total FOS Concentration, sucrose levels displayed p-
values of 0.0085 at 24h, 0.028 at 48h and 0.046 at 72h
CONCLUSION
These results are showing that this process is very promising for the FOS industry considering
that the conditions still not have optimized and more research has been done to evaluate and
optimize this process. This in turn can provide a focus for effective use of Aureobasidium
pullulans to synthesize fructooligosaccharide through bioconversion using entire cells to obtain
the product by direct manner.
REFERENCES
[1] Gibson, G.R., Roberfroid, M. B. 1995. Dietary modulations of the human colonic microbiota –
introduction and concept of prebiotics. Journal of Nutrition, 125: 1401-1412.
[2] Yun, J.W. 1996. Fructooligosaccharides—Occurence, preparation, and application. Enzyme Microbiol
Technology,19, 107-117.
[3] Rodrigues MI, Iemma AF (2005) Planejamento de experimentos e otimização de processos: uma
estratégia sequêncial de plane-jamentos. Editora Casa do Pão, Campinas, Brazil.

2108
Obtaining and characterization of mango peel powder and its use as a source of fiber and

a functional ingredient in natural yogurt

C. Ruiz, C. Ramírez, C. Gutiérrez de Piñeresc, M. Ángulo, J. Hedreira

a
Carlos Alberto Ruiz Galván, Valledupar, Colombia (carlosruizg67@yahoo.es)
b
Carlos Arturo Ramírez Guzmán, Valledupar, Colombia (ramirezc16@hotmail.com)
c
Carlos Alberto Gutiérrez de Piñeres (walframio@yahoo.com)

INTRODUCTION
The mango (Mangifera indica L) is a tropical fruit of high production and consumption in
Valledupar and the entire Atlantic Coast, which is characterized by a high nutritional value.
However, its use is limited to the pulp. In this research, it was decided to use ripe mango peel
variety yarn by making a powder for use dual functional ingredient and additive, due to its high
content of fiber and phytochemicals. The powder was obtained from mango variety yarn using
the following processes: selection, washing, peeling, drying, grinding, packaging and heavy.
Among the physicochemical characteristics are: moisture 6.24%, 2.24% fat, 2.82% protein,
10.35% neutral fiber, ash 4.23%, 2.82% citric acid, pH 4.69, ascorbic acid 0.032%, reducing
sugars 14.25% and 12.80% starch.

MATERIALS & METHODS

The powder was obtained from mango variety yarn through processes of selection, washing,
peeling, drying, grinding, packaging and heavy. Physicochemical analysis were performed
respectively: Moisture, Fat, Crude, Protein, Fiber neutral, Ash, Citric Acid, pH, Ascorbic acid,
Reducing sugar, Starch.
We used healthy and ripe, with its characteristic color (yellow), the variety of yarn, from the
estate of the Caribbean Biotech Center, located in the city of Valledupar, Colombia, during the
period of April and May 2010. The degree of maturity was shown by the color, fruit firmness
to the touch, and was considered at the same time the absence of physical damage and plant.
Natural yogurt was elaborated within the parameters of asepsis and hygiene and was added
different concentrations: 3%, 5%, 7% and 10% of mango peel powder, to note that the
physicochemical and sensory characteristics had the yogurt with the addition of this dust.

RESULTS & DISCUSSION

Mango powder obtained was analyzed in the laboratory analysis of food Biotechnology Center
of the Caribbean, the following results:

11th International Congress on Engineering and Food - Athens, Greece, 2011 2109
 Table 1. Physicochemical analysis of mango
Analyses Results
Moisture 6,24%
Fat 2,24%
Crude Protein 2,82%
Fiber neutral 10,35%
Ash 4,23%
Citric Acid 2,82%
pH 4,69
Ascorbic acid 0,032%
Reducing sugar 14,25%
Starch 12,80%
Source: Authors

The yield of mango peel powder after all processing, grinding and sieving was 40%. The
natural yogurt made with addition of 10% of mango peel powder before inoculation, showed a
good texture, flavor and color characteristic handle and had a shelf life of 1 month without
adding preservatives.

CONCLUSION
The mango peel powder has a yield of 40%, has very good sensory characteristics, such as taste
and characteristic yellow color, showed a 70% solubility in yogurt also providing its
characteristic smell and color of mango, indicating that in addition to behaving functional
ingredient due to its high fiber content also acts as a flavoring and coloring. The significant
content of beta carotene in the peel of mango yogurt offers a natural preservative system, since
this was a useful life of 30 days.

REFERENCES
[1] Ajila, C., Leelavathi, K. and Prasada, U. 2007. Improvements of dietary fiber content and antioxidant
properties in soft dough biscuits with the incorporation of mango peel powder. Journal of Cereal
Science, 1 – 8. This article in press.
[2] A.O.A.C. 1995. Official Methods of Analysis. 16th Ed. Association of Official Analytical Chemists.
Arlington, VA, USA.
[3] Campbell L, Keteisen S and Antenuci R. 1994. Formulating oatmeal cookies with calories sparing
ingredients. Food Technology, 48 (5): 101-105.
[4] Chau, C., Huang, F. 2003. Comparison of the chemical composition and physicochemical properties
of different fibers prepared from the peel of Citrus sinensis L. Cv. Liucheng. Journal of Agricultural
and Food Chemistry, 51: 2615 - 2618.
[5] Vega y León, S., Vega y Rojo A., Pérez F. y Coronado, H. 2002. Alimentos e ingredientes
funcionales. Industria Alimentaria, 24 (1): 13-18.

2110
Influence of gamma radiation on sprouting inhibition of the rhizomes and on the quality

of turmeric

Lucia Peret-Almeida & Maria Beatriz A. Gloria

LBqA – Laboratório de Bioquímica de Alimentos, Faculdade de Farmácia, UFMG, Av.

Antônio Carlos, 6627, CEP 31270-901, Belo Horizonte, MG, Brasil (mbeatriz@ufmg.br)

INTRODUCTION
Interest in turmeric has increased significantly recently. Turmeric has many nutritional and
functional properties, among them, antimicrobial, anti-inflammatory, antioxidant and
anticarcinogenic. It is widely used in Indian and Chinese medicine, for the treatment of liver
and gallbladder disorders, anorexia, rheumatism and sinusitis [1]. Furthermore, turmeric has
been widely used in the food industry as a natural colorant [4]. Immediately after harvest,
turmeric rhizomes have to be processed to avoid sprouting, which can cause a decrease on
quality [5]. By inhibiting sprouting of the rhizomes, it is possible to increase its shelf life and
to warrantee the quality of turmeric powder. Several chemicals have been used to inhibit
germination. However, there is concern regarding the accumulation of residues in the rhizome.
Irradiation is an interesting alternative to prevent sprouting. It offers advantages, among them,
short application time, no temperature change during the process, and lower toxicological risk
[2]. The objective of this study was to investigate the influence of gamma radiation on the
prevention of sprouting and on the quality of the turmeric powder during storage.

MATERIALS & METHODS

The rhizomes were washed and classified according to size. The samples were submitted to
gamma radiation at doses of 0.0, 0.05, 0.10 and 0.15 kGy in a gamma cell chamber (Atomic
Energy of Canada Limited, Canada) from CDTN, CNEN, UFMG. The source was Cobalt 60
with a potential of 0.0341 kGy/h. After irradiation, the rhizomes were stored at 26 ± 1 ºC at
85% relative humidity for up to 135 days. In 45 day intervals samples were collected and
evaluated for the presence and number of sprouts. The samples were also processed into
turmeric powder which was analyzed for curcumin and color characteristics [3]. The results
were submitted to analysis of variance and the means were compared by the Duncan test at 5%
probability

RESULTS & DISCUSSION

During storage of the irradiated rhizomes at 26 ± 1 ºC and 85% RH, the presence of sprouts
was observed on the 45th storage day in controls and also in samples submitted to 0.05 kGy.
However, when irradiated with 0.10 kGy, sprouts were only observed on the 90th storage day,
whereas no sprouts were observed on rhizomes irradiated with 0.15 kGy up to 135 storage
days. During storage of the rhizomes (control), there was a significant loss of curcumin
pigments on 135 days of storage (20% loss). The irradiation doses used did not affect the loss

11th International Congress on Engineering and Food - Athens, Greece, 2011 2111
of curcumin, which followed changes similar to controls (Figure 1). Irradiation of the
rhizomes did not affect the color characteristics of the rhizomes. Storage time did not affect
the color parameters L* (lightness) values, chroma and hue of the turmeric. However, there
was a decrease on a* (intensity of red) and b* (intensity of yellow) values with storage time,
independent on the irradiation dose used.

Figure 1. Loss of curcumin (%) during storage of turmeric submitted to gamma radiation at 0.00, 0.05,
0.10 and 0.15 kGy for up to 135 days.

CONCLUSION
Gamma radiation at 0.15 kGy was effective in the prevention of sprouting for 135 days of
storage at 26 ± 1 ºC and 85% RH, whereas doses of 0.10 kGy prevented sprouting for 90th day.
Irradiation did not affect significantly the levels of curcumin and the CIE L*a*b* color
characteristics. Therefore, gamma radiation at 0.15 kGy can be used to delay sprouting of the
rhizome without affecting turmeric quality.

REFERENCES
[1] Ammon H.P.T. & Wahl, M.A. 1991. Pharmacology of Curcuma longa. Planta Medica, 57, 1-7.
[2] Dhanya R., Mishra B.B., Khaleel K.M. & Cheruth A.J. 2009. Shelf life Extension of Fresh Turmeric
(Curcuma longa L.) using Gamma Radiation. Radiation Physics and Chemistry, 78(9), 791-795.
[3] Péret-Almeida L., Cherubino A.P.F., Alves R.J., Dufossé, L. & Gloria M.B.A. 2005. Separation and
Determination of the Physic-chemical Characteristics of Curcumin, Demethoxicurcumin and
Bidesmetoxicurcumin. Food Research International, 38, 1039-1044.
[4] Safford R.J. & Goodwin B.F.J. 1985.Immunological Studies on Tartrazine, International Archives of
Allergy and applied Immunology, 77(3), 331-336.
[5] Van Kooij J.G. 1986. International Trends in Uses of Food Irradiation. Food Reviews International,
2(1), 1-18.

2112
 Antioxidant dyes and pigment extraction using a home-made pressurized solvent
extraction system

Diego T. Santos, Carolina L. C. Albuquerque, M. Angela A. Meireles

LASEFI/DEA/FEA (School of Food Engineering)/UNICAMP (University of Campinas) – R. Monteiro

Lobato, 80; 13083-862, Campinas, SP, Brazil (meireles@fea.unicamp.br)

INTRODUCTION
Increasing reports of health hazards and toxicity of synthetic pigments are driving the food
industry towards application of natural colorants in an increasing number of processed food
products. Commonly, conventional extraction methods are used to extract these compounds
from natural sources, nevertheless, these methods are, in general, time and solvent consuming
and may promote the degradation of these compounds.
In order to overcome these drawbacks short time extraction conditions using pressurized
solvent methods, such as Supercritical Fluid Extraction (SFE) and Pressurized Liquid
Extraction (PLE) methods have been used successfully to obtain antioxidant pigments-rich
extracts [1, 2].
Carotenoids and anthocyanins are two of the most widely used dyes in food, pharmaceutical
and cosmetic industries.
In this work we designed and built a home-made pressurized solvent extraction system, in
which pure supercritical CO2 (SFE) and GRAS solvent (PLE) can be used, independently. SFE
was used for obtaining Annatto seed extract and PLE for Jabuticaba skin extract. Home-made
SFE results were compared to those obtained using a commercial SFE using the same
processing conditions. Moreover, fractionated extractions of Jabuticaba skins were performed
in two steps: i) a first step (PLE), wherein pressurized ethanol was used in order to extract
polar compounds like anthocyanin pigments; ii) a second step, wherein supercritical CO2 was
used in order to recovery low polarity CO2-soluble compounds. The chemical compositions of
both extracts were characterized.

MATERIALS & METHODS

Extraction Procedures
The cell containing a 4.51 g of plant material (whole Annatto seeds or dried/cut Jabuticaba
skins) was first loaded into the extraction cell, filled with extraction solvent (CO2 or ethanol)
and then pressurized. During 5 min the sample was placed in the heating system to ensure that
the extraction cell will be at the desired temperature at the filling and pressurization procedure.
After pressurization, the sample with pressurized solvent was kept statically at the desired
pressure for a desired time (static extraction time). Thereafter, carefully the blocking and
micrometric valve were opened keeping the pressure constant to the desired flow rate being the
extraction cell rinsed with fresh extraction solvent during a certain time (dynamic extraction
time).
For SFE process, liquid CO2 99.9 % (Gama Gases Especiais Ltda., Campinas, Brazil) was fed
from the cylinder through a thermostatic bath at -10 °C to ensure the liquefaction of the gas and
to prevent cavitation, and then it was pumped by the CO2 pump to the extraction cell
containing Annatto seeds or Jabuticaba skins previously extracted with pressurized ethanol.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2113
RESULTS & DISCUSSION
Figure 1 shows that the behavior of the extraction curves was very similar, independently of
the system used. Nobre et al. [3] using a similar SFE apparatus observed analogous extraction
curves for pigment extraction using also pure supercritical CO2 and whole Annatto seeds.

Figure 1. Recovery of pigments from Annatto seeds as a function of time using different SFE
systems/configurations: Ŷ Commercial SFE; Ƒ Home-made SFE.

Fractionated extractions of Jabuticaba skins were performed in two steps: i) a first step (PLE),
wherein pressurized ethanol was used in order to extract polar compounds like anthocyanin
pigments; ii) a second step, wherein supercritical CO2 was used in order to recovery low
polarity CO2-soluble compounds.
The chemical composition the PLE extract was characterized to the present of anthocyanins.
Visually the extract solution presented a purple color. The anthocyanin content of the PLE
extract was 2.489 ± 0.544 mg cyanidin-3-glucoside/g dry material. Otherwise, during the re-
extraction of the Jabuticaba skins previously extracted by PLE using supercritical CO2 an
extract presenting a yellow-green color, with insignificant anthocyanin content, was obtained at
the processing conditions employed.

CONCLUSION
In this work we validated a home-made pressurized solvent extraction system that can be used
for Supercritical Fluid Extraction (SFE) and Pressurized Liquid Extraction (PLE) processes,
independently, using Annatto seed and Jabuticaba skin as model plant materials.
Fractionated extractions of Jabuticaba skins were performed using our apparatus with success,
producing two valuable extracts.

REFERENCES
[1] Silva G.F., Gamarra F.M.C., Oliveira A.L. & Cabral F. A. 2008. Extraction of bixin from annatto
seeds using supercritical carbon dioxide.Brazilian Journal of Chemical Engineering, 25, 419-426.
[2] Arapitsas P. & Turner C. 2008. Pressurized solvent extraction and monolithic column-HPLC/DAD
analysis of anthocyanins in red cabbage. Talanta, 74, 1218-1223.
[3] Nobre B.P., Mendes R.L., Queiroz E.M., Pessoa F.L.P., Coelho J.P. & Palavra A.F. 2006.
Supercritical carbon dioxide extraction of pigments from bixa orellana seeds (experiments and
modeling). Brazilian Journal of Chemical Engineering, 23, 251-258.

2114
 Comparative study of the physicochemical characteristics of an economic Buffalo
(Bubalus bubalis) meat product and an economic Beef (Bos indicus) meat product with
incorporation of bovine hemoglobin in powder in both formulations

J.F. Rey1, C.L. Martínez2, A. Urrea3

1
Profesor del Programa de Ingeniería de Alimentos de la Universidad de la Salle, Bogotá, Colombia
(javierrey79@yahoo.com )
2
Egresada del Programa de Ingeniería de Alimentos de la Universidad de la Salle, Bogotá, Colombia
(anifmc@hotmail.com)
3
Egresada del Programa de Ingeniería de Alimentos de la Universidad de la Salle, Bogotá, Colombia
(ing.adrianaurrea@gmail.com)

INTRODUCTION
Low dietary intake of bioavailable iron is the major cause of iron deficiency and anemia deficiency. The
food-based approaches to increase iron intake through fortification food and dietary diversification are
also important strategies to prevent low levels of iron (WHO, 2004). Several studies have demonstrated
that buffalo meat has higher iron proportions than other species, protein content and low fat values
becoming a raw material of high potential for industry (Cedres, 2003).

MATERIALS AND METHODS

The product was made following sausage meat emulsions formulation (Ranken 2003). Were designed
and developed three buffalo meat sausages (B) with 200 mg / kg of hemoglobin
of hemoglobin (range selected from Aleppo research and Duke, 2009), and three beef (R) with the same
hemoglobin concentration. To evaluate the physicochemical characteristics of the sausages, were
tested: Total ash (NTC 1668), Humidity (NTC 1663), Protein (NTC 1556), Total fat (NTC 1162) iron
AOAC 944.02. Also, difference between the samples with bilateral Dunet test (ICS 95%). The
statistical comparison was performed with a standard deviation confidence interval of 95% was used.
The analysis of variance (ANOVA) was performed with significance (P <0.05), means were compared
using the Tukey test (P <0.05 and showed the difference between the samples, performing a test Dunnet
bilateral (95% ICS).

RESULTS AND DISCUSSION

The average value for physicochemical and iron content of fortified meat products with
dried bovine hemoglobin are shown in Table 1.
The statistical treatment showed no significant differences for the values of fat (%)
(R = 14.2352 and B = 9.7446), moisture (%) (R = 64.9434 and B = 65.3612) Protein (%)
(R = 13.5347 and B = 14.0219) and pH (B = 7.01 and R = 6.71). Also a significant difference for ash (%)
(R = 1.7572 and B = 2.8013) and iron (B = 2.82 mg/kg and R = 2.52 mg/kg) the data obtained exceed the
minimum recommendations for minimum intake of iron in Colombia (1,184 mg / kg, ICBF) and Latin
American food (1.1 mg/ kg, FAO).
The pH values for beef and buffalo are consistent with those stipulated by Ellas et al
(2007) where the sausage meat range is 5.8 to 7.0.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2115
 Table 1. Physicochemical characterization of B and R samples
Samples
Variable
R B
Fat (%) 14,2352 ± 12,312 9,7446 ± 3,916
Moisture (%) 64,9434 ± 8,045 65,3612 ± 4,036
Ash (%) 1,7572 ± 0,476 2,8013 ± 0,274
Protein (%) 13,5347 ± 6,98 14,0219 ± 3,419
pH 7,01466 ± 0,273 6,7163 ± 0,273
Iron (mg/100 g) 2,52 ± 0,125 2,82 ± 0,236

For ash, values there was a significant difference between the two results, the product made
from buffalo meat, has more minerals, and has higher quantities of iron.
Statically was determined that the sample B is significantly different from the sample A,
showing a higher iron content of the sample B in relation to R.
Also, data obtained in both samples exceed the limits from Colombia and Latin American typical food,
with a portion of 100mg, both products. The beef sausage, would provide
approximately 36% more than 100g of conventional product per serving g (levels of absorption
into the body are not considered).
The quantity of absorption is variable and depends on various factors such as individual's age, sex, type of
food according to FAO (2007). Buffalo meat has 53% more iron than required by current regulations.
Buffalo product (B) was selected with the best physicochemical and nutritional characteristics, as
well as a partial solution to low iron consumption in the population. According to
the FAO minimum daily intake should be at least 6 mg.

CONCLUSION
Aviable Iron contents are significantly affected with the addition of bovine hemoglobin (200mg/Kg) to an
economic sausage made by select quality cuts from beef and buffalo, especially contents of the buffalo
one.
Protein levels were increased and saturated fat decreased in relation with beef meet, making it a
candidate for industrial economic products like "healthy ", "high iron" or "low fat".
Buffalo product presents healthy physicochemical characteristics respect to Beef product, especially in
terms of protein, fat and iron contributions against securities FAO and ICBF requirements.

REFERENCES
CEDRES, J. (2002). Chemical composition and physical characteristics of buffalo meat extensively bred in the
province of Formosa. UNNE. Faculty of veterinary science. Corrientes.
ELLAS et al. (2007). Effect of different binders on the quality of enrobed buffalo meat cutlets and their shelf life at
refrigeration storage (4 ± 1 ºC). Meat Science 75 (2007) p. 451–459.
OMS. (2004). Consultada el 05 de Febrero de 2010. Anemia as a center of attention. Available
at:http://www.paho.org/Spanish/AD/FCH/NU/OMS04_Anemia.pdf.Accessed on 05 February 2010
CARRASCO, R. (2008). Evaluation of powder of hemoglobin as a substitute of synthetic red dye Ponceau 4R in
the development of economic sausage. (pp. 1-106). La Salle University. Food Engineering Program. Bogotá.
FAO. Table of food composition Sausage F451 Latin America. Available
online: http://www.rlc.fao.org/es/bases/alimento/print.asp?dd=3259.Accessed May 20 2010:
GUERRERO, R. (2006). The impact of iron deficiency on psychomotor development. Available
at:http://www.utp.edu.co/facies/educacioncontinua/maternoinfantil/EL_IMPACTO_DE_LA_DEFICIENCIA_DE
_HIERRO_EN_EL_DESARROLLO.pdf.Accessed May 5, 2010.

2116
 Production of Turkish delight (lokum) with its additives and quality

Ali Batu
Tunceli University, Engineering Faculty, Food Engineering Department, 62100 Tunceli Turkey
(ali_batu@hotmail.com)

INTRODUCTION
In this study, a brief evaluation will be made on Turkish delight (lokum) that is a starch jelly a
traditional Turkish food product and has importance for the Turkey. The functions of the main
ingredients such as sugar, starch and water on the overall lokum quality will be discussed.
Then production problems and some additives have been used in lokum production will be
indicated. Coconut, peanut, pistachio, almond and etc. are used nuts in Lokum production. The
origin of Lokum dates back to the time of the Ottoman. Beginning of production of lokum is
estimated at 15th century and its production is reached today’s form at 19th century. In 19th
century lokum was brought to England by a English tourist, then it called “Turkish delight” in
Europe and “Lokoum” in France and Balkans, lokumania in Greece and in Cyprus, and then
lokum took place in international candy literature. Lokum is produced by mixing of sugar, corn
starch and watering certain fraction and fruits or nuts are added to the mixture. This mixture is
heated for certain time at certain temperature in open vessel or steam jacketed tank with
agitator. Heating time changes 1 to 2 hours according to vessel’s type. Hot lokum fluid is
dripped in wooden table or steel tray, which some starch is on, with lower edges, after
sprinkling some starch lokum are cut as small particles. Powder sugar is added on this lokum
particles and they are stored and sold. Keywords: Acid, sugar, lokum, modified starch,
confectionary, Turkish delight

THE HISTORY AND INVENTION OF LOKUM

An old Turkish aphorism tells one to “eat sweetly and speak sweetly”. The origin of Lokum
dates back to the time of the Ottoman. Beginning of production of lokum is estimated at 15th
century and its production is reached today’s form at 19th century [1]. Lokum is a sugar based
jelly-like confection containing starch with the gel former. According to legend, Turkish
Delight, one of the oldest and most delectable sweets in the world, was created some 500 years
ago when a Turkish sultan asked his confectioner to produce something sweet to keep on his
family and friends [2]. Lokum was unveiled to the west in the 19th century. During his travels
to Istanbul, an unknown British traveler became very fond of the Turkish delicacies, purchased
cases of lokoum and he shipped them to Britain under the name Turkish delight. Today,
Lokum remains the sweet of choice in many Turkish homes and enjoyed worldwide [3].
Picasso used to eat lokum on a daily basis for concentration on his work while Winston
Churchill and Napoleon's favorite Turkish Delight was with pistachio filling [2]. One major
commercial producer in the Northwestern U.S. is Liberty Orchards, founded by Armenian
immigrants, which markets the candy under the name "Aplets and Cotlets" and "Fruit
Delights." It is also the basic foundation of the Big Turk chocolate bar commonly found in
Canada. Another North American company is the Bayco is the manufacturer of authentic
Lokum in North America [4].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2117
THE RAW MATERIALS
Sugar: Sugar is the one of the most important raw material in Lokum production. Starch is the
major component of grains and a common ingredient used in the food industry. Understanding
the relationship between the structure of starch and rheological properties of lokum will
improve the ability to manipulate texture and could result in identification and development of
lines and mutants of starch with abilities to resist breakdown. Starch used in lokum production
as a basic raw material has important quality criterions [5]. Acid. Citric and tartaric acid are
used in Lokum production in Turkey. Manufacturers often determine acid amount to be added
by themselves. At the end of the researches, using 5g tartaric acid or 3g citric acid results the
godd quality of lokum. Water is also the one of the most important raw material effecting
quality after sugar. Especially soft water increases quality, on the contrary water with high lime
content destroys the structure [6].

LOKUM PRODUCTION PROCESS IN TURKEY

Preparing sugar syrup and starch milk: First of all, syrup is made from sugar with enough
water to melt it. Practically, this mix is called as starch milk. Starch milk is added to boiling
sugar syrup. In this step of boiling acid is added to invert sugar [7]. Cooking: Once the water
has boiled the sugar is added and the solution is boiled for an hour, stirred continuously by an
electric paddle. Next the starch is added, and the mixture brought back to the boil, for another
five to six hours until it is smooth and shiny. The mixture is allowed to cool for a while, before
the various flavors are added. After the flavorings have been carefully added, the mixture is
poured into large wooden trays to set. About five hours later it is ready to be cut into squares,
liberally dusted with icing sugar and packed into small boxes lined with greaseproof paper
ready for selling. Lokum production process is given [3]. Cooling: Lokum is cooled in room
conditions for 24hrs after pouring. Recently cooling may be done in 3-4 hours with water
cooling systems. Both cooling methods and time may lower quality. Cooled Lokum is put on
cutting tables. In our country, cutting of Lokum is done by hand or machine. Sugar powder or
coconut is found on table. Cut Lokum’s are fitted in boxes in desired weight and transported to
storage rooms to sell. Generally, Lokum which is sold in Turkey is packed in 5kg wooden
boxes. To prevent sticking of lokum to box polyethylene, greasy and waxy paper are used [7].

REFERENCES
[1] Batu, A. and KÕrmacÕ, B. 2009. Production of Turkish delight (lokum). Food Research International
42 (2009) 1–7
[2] Anon, A. http://pedia.nodeworks.com/L/LO/LOK/Lokum
[3] Batu, B. 2006. Türk lokumu Üretim Tekni÷i ve Kalitesi. GÕda Teknolojisi Elektronik Dergisi. 2006
(1) 35-46.
[4] Anonymous, 2006. Turkish Delight, Bayco confectionery Inc., Canada, Manufacturer' Of Authentic
Turkish Delight http://www.turkish-delight.com/
[5] SaldamlÕ, ø. 1998. GÕda KimyasÕ. Karbonhidratlar. 2. Bölüm. S:37-105. Hacettepe Üniversitesi.ISBN
No: 9758339001, Ankara
[6] Gönül, M. 1985. Türk Lokumu YapÕm Tekni÷i Üzerine AraútÕrmalar. 1. BaskÕ. Ege Mühendislik
Fakültesi. Ders KitaplarÕ YayÕn No: 8, Bornova, øzmir
[7] Anonymous, E. ArÕ Lokum. http://www.arilokum.com.tr/lokum_imalat.asp

2118
 Effect of fermented okara (bean curd lees) intake on TNCB
(2, 4, 6-trinitrochlorobenzene)-induced chronic dermatitis in NC/Nga mice

Toshiki Enomotoa, Masato Nishia, Florin Barlaa, Nana Murataa, Hiroshi Matsuib, Hidehiko Kumagaib,
Harumi Takec, Toshihide Michihatac, Shizuo Nakamurac, Masao Kawashimad, Eiji Fujiharad

a
Department of Food Science, Ishikawa Prefectural University, Nonoichi, Ishikawa 921-8836, Japan
(enomoto@ishikawa-pu.ac.jp)
b
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi,
Ishikawa 921-8836, Japan (hidekuma@ishikawa-pu.ac.jp)
c
Laboratory of Food Processing, Industrial Research Institute of Ishikawa, Kanazawa, Ishikawa 920-
8023, Japan (take@irii.jp)
d
Habutaetoufu Co.,Ltd, Kanazawa, Ishikawa 921-8054, Japan (fujihara@habutae.co.jp)

INTRODUCTION
Atopic dermatitis (AD) is a complex eczematous skin disease accompanied by severe itching
and frequently repeated episodes. The prevalence of AD has increased steadily in recent
decades. Anti-allergic functional food has been attracting attention, and lactic acid bacteria has
been one of the main focus of reports.In East Asian countries, tofu is a popular food. Okara is
produced as a by-product of tofu manufacturing. Although okara is a rich source of nutrient
which might be beneficial for human health, the most of it is disposed and unused. Therefore
the development of a processing way for utilization is expected. The aim of the study was to
investigate the effect of fermented okara intake on TNCB (2, 4, 6-trinitrochlorobenzene)-
induced chronic dermatitis in NC/Nga mice.

MATERIALS & METHODS

Four weeks-old NC/Nga male mice which have a predisposition to spontaneously develop AD-
like dermatitis were purchased from Japan SLC, Inc. Control feed, MF was purchased from
Oriental Yeast Co. Ltd. Water and food were available ad libitum. Next, they were randomly
divided into three groups (n=6/group). The control group was fed a standard MF and the
experimental groups were fed a mixture of the standard MF plus fermented okara to give a
final fermented okara concentration of 1% (1% FO) and 2% (2% FO), respectively. Then, mice
were sensitized initially, at the beginning of experiment, with about 20 μl of TNCB 5%
solution onto ventral zone and feet. The following weeks, the TNCB 1% solution was applied
to sensitized ears and scruff zone once per week. The animals were fed these diets for 5 weeks
and ear thickness was measured with an electronic calliper every week just before sensitization.
The whole blood was collected from inferior vena cava, under anaesthesia with diethyl ether,
and placed immediately into ice. Serum samples were obtained by centrifugation, and were
used to quantify the level of cytokine.

RESULTS & DISCUSSION

Ear thickness of mice was measured weekly and an increase in the thickness was observed in
all groups. However, mice in the fermented okara-administrated groups had a significantly
smaller ear thickness compared with the control group. Oral administration of fermented okara
to NC/Nga mice inhibited the development of AD-like skin lesions based on the total skin

11th International Congress on Engineering and Food - Athens, Greece, 2011 2119
severity scores, while the scores considerably increased in the control group (Fig.1A and B).
These results suggest that the symptoms of mice fed a diet containing fermented okara were
markedly reduced. Furthermore, the IFN- Ȗ level of serum was not affected by fermented okara
administration in both mice groups but in contrast the IL-4 level was significantly decreased,
indicating that the Th1/Th2 balance in fermented okara groups was higher than the control.

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A
㻜㻚㻢㻡㻌 a
a
a a
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㻕
㼙
㼙
㻔㻌 a
㼟㼟㻜㻚㻠㻡㻌 a
㼑
㼚㼗 a
b b
a
㼕㼏㼔 a a b
b b
㼠㻌 㻜㻚㻟㻡㻌 a a b b b b
㼞 b b b b
㼍 b b b
㻱 b b
㻜㻚㻞㻡㻌

㻜㻚㻝㻡㻌
㻝㻞㻚㻜㻌
㻝㻝㻚㻜㻌 B
a a
㻝㻜㻚㻜㻌 a
㻥㻚㻜㻌 a

㻤㻚㻜㻌 a
㼑㼞 㻣㻚㻜㻌 a
㼏㼛
㼘㻌㼟 㻢㻚㻜㻌 b b
㼍 b b
㼏㼕 㻡㻚㻜㻌 b b b
㼘㼕㼚 㻠㻚㻜㻌 b b b
㻯 a b
a
㻟㻚㻜㻌
a b
㻞㻚㻜㻌
㻝㻚㻜㻌 b c
b
㻜㻚㻜㻌
㻜 㻡㻌 㻝㻞㻌 㻝㻥㻌 㻞㻢㻌 㻟㻟㻌 㻠㻜㻌 㻠㻤㻌 㻡㻢㻌 㻢㻝㻌 㻢㻥㻌 㻣㻡㻌
㻰㼍㼥㼟 㼍㼒㼠㼑㼞 㼟㼑㼚㼟㼕㼠㼕㼦㼍㼠㼕㼛㼚

Figure 1. Effect of okara administration on sensitized ears thickness (A) and on the total clinical score
(B) induced by TNCB application in NC/Nga mice. Results are expressed as the meansSD of six mice
per group. Means with the different letters are significantly different at the 5% level (Tukey’s multiple
range test). ‫ە‬:control diet, ‫ڸ‬: 1% FO diet, ‫ڦ‬: 2% FO diet

CONCLUSION
From the results, fermented okara exerts an anti-allergic action through suppression of Th2-
type immune response. Thus, okara fermented by Bacillus coagulans might be effective dietary
supplement for the prevention of AD.

REFERENCES
[1] Matsumoto, K., Watanabe, Y. and Yokoyama, S. 2007. Okara, soybean residue, prevents obesity in a
diet-induced murine obesity model, Biosci. Biotechnol. Biochem., 71, 720-727.
[2] Prestamo, G., Ruperez, P., Espinosa-Martos, I., Villanueva, M. J. and Lastuncion, M.A. 2007. The
effects of okara on rat growth, cecal fermentation, and serum lipids, Eur.Food Res.Technol., 225,
925-928.
[3] Kitagai, H., Fujisawa, S., Watanabe, K. and Shiohara, T. 1995. Immediate-type hypersensitivity
response followed by a late reaction is induced by repeated epicutaneous application of contact
sensitizing agents in mice, J. Invest. Dermatol., 105, 749-755.

2120
 Development of a dehydrated and laminated probiotic product with B. infantis and L.

acidophilus using goat sweet whey

Grissel Trujillo de Santiagoa, Carlos Sáenz Collinsb, Cecilia Rojas de Gantec

a
Tecnológico de Monterrey, Monterrey, Nuevo León, México. (A00279913@itesm.mx)
b
Tecnológico de Monterrey, Monterrey, Nuevo León, México. (A00802349@itesm.mx)
c
Tecnológico de Monterrey, Monterrey, Nuevo León, México. (crd@itesm.mx)

INTRODUCTION
To promote the health benefits, the International Dairy Federation (IDF) suggests that probiotic
products must have a minimum concentration of 107CFU/g [1]. Besides, whey is a by-product
remainder from cheese manufacture that has high potential to be transformed into a surplus
good, such as probiotic products. The current work presents the development of a new product
consisting of a dehydrated and laminated probiotic dairy product that employs a by-product of
cheese manufacture and avoids the high energy costs of cold chain.

MATERIALS & METHODS

The strains L. acidophilus and B. infantis were reactivated in MRS broth and then adapted to
pasteurized goat’s sweet whey (GSW) through successive transfers until lag phase was
eliminated. Incubation took place in a vacuum incubator (Sheldon Manufacturing Inc.,
Cornelius, OR) at 38± 2°C and a vacuum of 20mm Hg. Growth kinetics were constructed
through counting plate using MRS agar (DIFCO) and the same incubation conditions
previously mentioned. Lactose consumption and lactic acid kinetics were determined through
HPLC- RI using an Aminex HPx87H column and H2SO4 as mobile phase. Inulin (in a 15% p/v
solution), resistant corn starch (RS) (in a 6% p/v suspension), and gelatin (in a 10% p/v
solution) were used as additives to get the DLP formulation. Two levels of each additive (8.33
and 16.66% of the final volume) were evaluated using a factorial 23 experimental design. Nine
judges evaluated three quality characteristics (film formation, homogeneity, and smoothness)
using a scale from 1 to 5. The process for obtaining the DLP consisted of five stages: (1)
fermentation of the GSW with B. infantis or L. acidophilus; (2) formulation of the fermented
GSW with inulin, RS, and gelatin; (3) pouring the formulation onto a non-stick pan; (4) drying
it in a convection oven with an airflow of 0.212±0.03m/min; and (5) dehydrating it in a
dessicator at ~0% relative humidity (RH) during 22h. Three drying temperatures (40, 55, and
70°C) were evaluated as a function of the bacterial survival. The bacterial viability was
determined through plate counts, using MRS agar and the same incubation conditions as
fermentation. The viability of microorganisms was also evaluated after dehydration at 0% RH.

RESULTS & DISCUSSION

No lag phase is observed on kinetics, as the strains were properly adapted to the GSW. Both
strains started the fermentation with 107CFU/ml and reached the next magnitude order at the
first hour of fermentation, achieving concentrations of 5.8×108CFU/ml and 5.2×108CFU/ml
for B. infantis and L. acidophilus, respectively. B. infantis maintained a logarithmic phase
during the first 6 hours and reach the stationary phase at 6 to 8.5 hours, followed by the dead

11th International Congress on Engineering and Food - Athens, Greece, 2011 2121
phase. L. acidophilus had logarithmic phase during the first 6.2 hours; from this point to 8.3 a
stationary phase occurred, followed by the dead phase. ȝmax and td were were 0.28h-1 and 2.25h,
respectively for B. infantis and 0.31h-1 and 2.22h for L. acidophilus. The greatest slopes of
lactose consumption and lactic acid production occurred from 2 to 4 hours (0.26g/L·h-1 and
0.35g/L·h-1 respectively for B. infantis, and 0.27g/L·h-1and 0.53g/L·h-1for L. acidophilus), the
same as in growth and pH kinetics. It can be inferred that at mid log phase the strains not only
had their maximum growth rate, but also presented an accelerated metabolic activity. Although
the reported optimum growth pH for B. infantis and L. acidophilus is 6-7 and 5.4-6.0
respectively [2], these strains were able to growth at lower pHs. Fermented GSW in the mid
log phase was used in the formulation to get DLP, because in this stage the bacteria are able to
produce “stress proteins” that allows them to create a homeostasis system to new conditions
[3]. The graphics derived from statistical analysis were used to choose the best formulation.
The formulation that achieved the highest averages of the three quality characteristics was the
one with the high levels of gelatin and inulin and low level of RS. High level of inulin was
always desired, as it impacts positively on every quality characteristic; low levels of RS
favored smoothness and film formation; gelatin seems to have a positive effect on homogeneity
at a high level. The feasibility of the process was highly determined by the influence of drying
conditions on the viability of probiotic bacteria. The concentration of viable probiotic bacteria
was higher than 107UFC/g after the drying in all cases studied; nevertheless, the highest
concentrations were achieved at 55±5°C for both strains After drying, a product with
9.47±1.2% of moisture was obtained; this moisture content conferred plasticity to the matrix
and allowed a proper detachment. A final dehydration step was necessary to decrease the
moisture content from 9.47% to 3.5% in order to get a dry-crispy laminated product; therefore,
after drying, the product was subjected to a 0% RH environment. During this process, the
concentration was diminished by 86.29±2.52% and 73.51±4.13% for B. infantis and L.
acidophilus, respectively, but enough concentration to develop probiotic effects remained in
the final products

CONCLUSION
A process to develop a DLP with viable probiotic bacteria (B. infantis and L. acidophilus) was
successfully achieved. Even when the probiotic bacteria were exposed to high temperatures
(55±5°C) and arid conditions (0%RH) during the process, the concentration of viable bacteria
remained above the minimum effective concentration (107CFU/g) in the final product. Besides
the technological functions of the ingredients used in the formulation, it is inferred that they
played an important role to protect the bacteria from hostile conditions. Further studies are
needed to evaluate the viability of probiotic bacteria during DLP shelf life.

REFERENCES
[1] Kailasapathy K. 2002. Microencapsulation of probiotic Bacteria: Technology and Potential
Applications. Current Issues in Intestinal Microbiology, 3(2), 39-48.
[2] Gomes A.M.P. & Malcata, F.X. 1999. Bifidobacterium spp. and Lactobacillus acidophilus: biological,
biochemical, technological and therapeutical properties relevant for use as probiotics. Trends in Food
Science and Technology, 10(4-5), 139-157.
[3] Shah N.P. 2000. Probiotic Bacteria: Selective Enumeration and Survival in Dairy Foods. Journal of
Dairy Science, 83(4), 894–907.

2122
 Lentil-based snacks: Structural and textural evaluation

Andriana Lazoua, Magdalini Krokidaa, Nikolaos Zogzasb, Vaios Karathanosc

a
Laboratory of Process Analysis and Design, School of Chemical Engineering, National Technical
University of Athens, 5 Iroon Polytechniou St., Zografou Campus, 15780 Athens, Greece
(alazou@central.ntua.gr, mkrok@chemeng.ntua.gr)
b
Laboratory of Food Engineering, Department of Food Technology, Technological Educational Institute
of Athens, Agiou Spyridonos St., 122 10, Egaleo, Athens, Greece (nzogzas@teiath.gr)
c
Department of Nutrition, Harokopio University, 70 El. Venizelou St., 17671 Kallithea, Athens, Greece
(vkarath@hua.gr)

INTRODUCTION
Grain legumes are important sources of food proteins and dietary fibers, as well as, basic
constituents of the Mediterranean diet [1]. Extrusion cooking technology is a versatile and efficient
method for the production of direct expanded snacks. Their acceptability by consumers depends on
both structural and textural characteristics [2]. Extrusion conditions and feed composition affect
structural and textural characteristics of extruded snacks. The extrusion behavior of protein-starch
systems has been reported previously [3], but there is still a lack of knowledge of extrudate
properties containing whole grain legumes.
The purpose of this study was to investigate the influence of extrusion conditions, along with raw
material composition, on textural (modulus of elasticity and number of peaks during compression),
structural (apparent density and expansion ratio) and sensory characteristics of the extruded corn-
lentil mixtures. The instrumental and sensory attributes were correlated and differentiations among
extruded snacks, produced under various conditions, were investigated. Simple empirical models
were developed for the prediction of the instrumental properties as a function of process conditions
and feed composition.

MATERIALS & METHODS

Lentil/corn flour mixtures (ratios of 0, 10, 30, and 50%) were used, while their moisture content was
adjusted (13, 16, and 19% wet basis). A co-rotating twin screw extruder with a die diameter of 3
mm was used, operated at 200 rpm. Extrusion temperature and feed rate were regulated, at 170, 200,
and 230϶C and 2.52, 4.86 and 6.84 kg/h, respectively. Apparent density of extrudates was
determined by the actual dimensions of the sample. Expansion ratio was calculated by the
diameters’ ratio of extrudate over die. Compression tests were performed at room temperature
(25϶C) using a UTM, with a 100N load cell. The extrudates cellular structure was investigated with
SEM, operated at 15V and x35 magnification. A ten-member trained panel participated in the
descriptive analysis of extrudates, while the evaluated attributes included diameter, crunchiness,
crispness, hardness, cohesiveness and melting.
The mathematical model used to predict instrumental properties, was a simple power model. The
proposed equation has the form:
nT,M nF,M nX,M
§ T · § F · § X · § 100C ·
nCM
,
(1)
M M0 ˜¨ ¸ ˜¨ ¸ ˜¨ ¸ ˜¨ ¸
©T0 ¹ © F0 ¹ © X0 ¹ ©100C0 ¹

11th International Congress on Engineering and Food - Athens, Greece, 2011 2123
where M is the property, M0 is the property at reference conditions, T is the extrusion temperature, F
is the feed rate, X is the feed moisture content and C is the lentil to corn ratio. The symbols with
zero indexes indicate the respective values at reference conditions. The reference conditions are
200°C temperature, 4.68 kg/h feed rate, 16 % wb feed moisture content and material ratio 30%.

RESULTS & DISCUSSION

The parameter estimates of instrumental properties, apparent density (ȡapp) expansion ratio
(Exp.Ratio), modulus of elasticity (E) and number of peaks during compression (N) are summarized
in Table 1.

Table 1. Results of parameters estimation of instrumental properties.

ȡapp Exp. Ratio E N
M0 0.146 ± 0.001 2.251 ± 0.014 1.151 ± 0.013 10.007±0.191
nT,M -0.537 ± 0.026 0.251 ± 0.021 -0.122 ± 0.035 -3.029±0.054
nF,M 0.075 ± 0.053 -0.282 ± 0.046 -0.061 ± 0.073 0.716±0.100
nX,M 0.741 ± 0.063 -1.086 ± 0.045 1.177 ± 0.083 -0.406±0.135
nC,M -0.615 ± 0.019 0.414 ± 0.025 -0.498 ± 0.025 0.100±0.045
R2 0.888 0.923 0.817 0.880

Structural properties were significantly affected by the process conditions (p<0.001). Apparent
density increased with feed rate, moisture content and mixture ratio, and decreased with
temperature. The expansion ratio exhibited the opposite behavior. Modulus of elasticity increased
with moisture content and material ratio and decreased with extrusion temperature and feed rate.
The number of peaks during compression decreased with extrusion temperature, material ratio and
moisture content. Sensory-evaluated structural and textural characteristics had similar behavior with
instrumental properties. The variations of structural properties were confirmed by macrostructure
observation using scanning electron microscopy. The correlation of instrumental and sensorial
characteristics showed that sensory diameter is related to expansion ratio, and crunchiness with the
number of peaks and expansion ratio. Furthermore the apparent density is correlated positively with
elasticity modulus. Simple empirical models were developed, which enable the prediction and
design of extruded snacks with acceptable quality characteristics.

CONCLUSION
Structural and textural properties of corn–lentil extrudates were investigated and related to process
conditions and material composition, proposing simple power models. The properties correlation
revealed critical relationships among instrumental and sensorial characteristics. It is possible to
predict sensory characteristics of extrudates, which are critical for consumer acceptance, via
instrumental measurements and using the proposed models.

REFERENCES
[1] Pérez-López, F.R., et al., 2009. Effects of the Mediterranean diet on longevity and age-related morbid
conditions. Maturitas, 64 (2), 67-79.
[2] Lazou, A., Krokida, M., &Tzia, C., 2010. Sensory properties and acceptability of corn and lentil
extruded puffs. Journal of Sensory Studies, 25 (6), 838-860.
[3] Ding, Q.-B., et al., 2006. The effect of extrusion conditions on the functional and physical properties
of wheat-based expanded snacks. Journal of Food Engineering, 73 (2), 142-148.

2124
 The study on SFLAB GanedenBC30 viability on baking products during storage

Chia-Ling Jaoa, Shih-Li Huangb, Shao-Chi Wua, Hsu Kuo-Chiangc

a
Department of Food Science and Technology, Tung-Fang Design University, Kaohsiung, Taiwan,
R.O.C. (cljao@mail.tf.edu.tw)
b
Department of baking technology and management, National Kaohsiung University of Hospitality and
Tourism, Kaohsiung, Taiwan, R.O.C. (hsl@mail.nkuht.edu.tw)
c
Department of Nutrition, China Medical University, Taichung, Taiwan, R.O.C.
(kchsu@mail.cmu.edu.tw)

INTRODUCTION
SFLAB (spore forming lactic acid producing bacteria) are a group of Gram-positive
bacteria, sharing characteristics common to the genera Bacillus and Lactobacillus. This group
includes B. coagulans, B. racemilacticus, B. laevolacticus, and members of the genus
Sporolactobacillus. Many factors make SFLAB good candidates for a probiotic use: (i) they
are easily cultured in ‘bulk’; (ii) they produce organic acids; and (iii) they posse the capacity to
sporulate [1]. In addition, in the spore form, SFLAB are more resistant to heat, which
facilitates the pelleting process used in the mass production of probiotic animal feeds. On the
other hand, SFLAB also have the advantage to have lower nutritional requirements, a good
resistance to different environmental stress [1]. SFLAB GanedenBC30 is the trademarked
name for the patented strain of probiotic bacteria, Bacillus (B.) coagulans GBI-30, 6086.
GanedenBC30 is a natural supplement that can help support your digestive health and maintain
your immune system [2].

MATERIALS & METHODS

For understand the GanedenBC30 used different way could affect their viability after baking.
The 8 different baking products were made from 0.5% GanedenBC30 that added in their dough
by two ways: (a) flour powder or (b) egg yolk. And then the (a) pH value, (b) titratable acidity,
(c) GanedenBC30 counts, and (d) viability GanedenBC30 of 8 different baking products were
determined after storage at 4oC for 0, 3, 6, 9, 12, 15 days, or 25oC for 0, 3, 6 days.

RESULTS & DISCUSSION

The 8 type’s dough had relatively lower pH value and rise after baking. As titratable acidity
of the 8 type’s dough appear relatively higher amount and descend after baking. However, pH
value and titratable acidity in the 8 baking products were maintaining the same without change
after 9 days under 4oC. On the other hand, the GanedenBC30 counts in the 8 baking products
were reducing than their raw dough contains GanedenBC30. Both storage at 4 and 25oC, the
results show the GanedenBC30 viability of baking products were decreasing with storage days,
and storage at 25oC had relatively decreasing on viability of GanedenBC30 than 4oC. And the
dough made by flour powder and baking that show higher GanedenBC30 viability than by egg

11th International Congress on Engineering and Food - Athens, Greece, 2011 2125
Table 1. pH values, titratable acidities (TA), GanedenBC30 counts (log CFU/mL), and viability of the
chrysanthemum cookies containing GanedenBC30 during storage under refrigeration temperature or at
room temperature
GanedenBC30 in Powder GanedenBC30 in Egg Yolk
pH log Viability pH log Viability
TA TA
value CFU/g (%) value CFU/g (%)
5.52 r 0.17 r 7.12 r 5.44 r 0.19 r 7.46 r
Raw Dough Day 100.0 100.0
0.01e 0.02a 0.03a 0.01d 0.02a 0.19a
5.87 r 0.11 r 6.96 r 5.61 r 0.13 r 6.74 r
0 68.9 18.3
0.01bc 0.02b 0.01b 0.01c 0.01b 0.01b
5.85 r 0.12 r 6.94 r 5.65 r 0.12 r 6.63 r
3 65.9 14.3
0.02cd 0.02b 0.02bc 0.02ab 0.01b 0.10bc
5.84 r 0.11 r 6.90 r 5.64 r 0.11 r 6.60 r
6 59.8 13.3
Refrigeration 0.02d 0.02b 0.01cd 0.01b 0.02b 0.03bc
Temperature 5.89 r 0.10 r 6.89 r 5.65 r 0.11 r 6.56 r
9 58.3 12.0
0.02b 0.02b 0.01d 0.01ab 0.02b 0.03bc
5.88 r 0.11 r 6.86 r 5.65 r 0.12 r 6.50 r
12 54.5 10.7
0.02bc 0.02b 0.02ef 0.02ab 0.01b 0.04bc
5.87 r 0.11 r 6.84 r 5.65 r 0.12 r 6.46 r
15 52.3 10.0
0.02bc 0.02b 0.03ef 0.02ab 0.01b 0.17cd
5.86 r 0.11 r 6.90 r 5.67 r 0.11 r 6.47 r
0 59.8 10.0
0.01cd 0.02b 0.01cd 0.01a 0.02b 0.12cd
Room 5.87 r 0.12 r 6.87 r 5.67 r 0.12 r 6.25 r
3 56.1 6.0
Temperature 0.02bc 0.02b 0.03de 0.02a 0.01b 0.07d
5.94 r 0.12 r 6.82 r 5.64 r 0.12 r 5.75 r
6 50.0 1.9
0.02a 0.02b 0.02f 0.02b 0.01b 0.07e

CONCLUSION
The GanedenBC30 are good candidates for baking products using, both in lactic acid
production and probiotic preparations for 8 test baking products.

REFERENCES
[1] Hyronimus B., Le Marrec C., Hadj Sassi A. & Deschamps A. 2000. Acid and bile tolerance of spore-
forming lactic acid bacteria. International Journal of Food Microbiology, 61(2-3): 193-197.
[2] Maathuis A.J.H., Keller D. & Farmer S. 2010. Survival and metabolic activity of the GanedenBC30
strain of Bacillus coagulans in a dynamic in vitro model of the stomach and small intestine.
Beneficial Microbes, 1(1): 31-36.

2126
 Formulation and characterization of biocompatible microemulsions as nutraceutics.

Aristotelis Xenakis, Vassiliki Papadimitriou, Theodore G. Sotiroudis

National Hellenic Research Foundation, Institute Of Biological Research & Biotechnology,
Athens, Greece. arisx@eie.gr

INTRODUCTION
Emulsion preparation in conventional food industries involves the application of energy to
mixtures of oil, water, and emulsifier that stabilizes the interfacial layer between the dispersed
and continuous phases. These macroemulsions are turbid, having droplet sizes ranging from
0.2 to 10 μm and may remain kinetically stable. Microemulsions, on the other hand, are
thermodynamically stable, transparent isotropic solutions with particle sizes ranging from 5 to
100 nm, and form spontaneously. Application of microemulsions in foods is limited by the
types of surfactants which are used. Many surfactants are not permissible in foods or may be
added at low levels. Microemulsions can be considered as delivery systems as they can
solubilize simultaneously hydrophilic, lipophilic and amphiphilic substances. Many studies in
the pharmaceutical and cosmetic fields reported enhanced solubilization of poorly soluble
compounds and improved bioavailability following incorporation into microemulsions. A
similar case could be the solubilization of compounds with bioactive capability isolated from
varying food sources.
In the present study the use of olive oil for the formulation of lecithin based microemulsions is
suggested. Virgin olive oil (VOO) contains a variety of minor components such as free fatty
acids, phospholipids (endogenous amphiphiles), polyphenols and partial glycerides [1]. Studies
from this laboratory have shown the presence of proteins and enzymes in VOO [2]. As a
consequence VOO itself can be considered a fine emulsion of a water phase in a continuous
non-polar phase [3].

MATERIALS & METHODS

Extra virgin olive oil samples (from olives of cv. Koroneiki) were from oil mills of Filiatra,
Greece. Refined olive oil was donated by Elais -Unilever S.A., Greece. Soybean lecithin
(Emulmetic 930) containing 92% phosphatidylcholine, was supplied from Lucas Meyer
Cosmetics SAS, France. 1-Propanol was from Merck, Darmstadt, Germany. High-purity water
was obtained from a Millipore Milli Q Plus system.
A typical olive oil based w/o microemulsion was prepared by mixing olive oil with a mixture
of lecithin/1-propanol (1:3) resulting a solution containing 85% w/w oil and 15% w/w
emulsifiers. Then appropriate amounts of water were added to obtain a clear reverse micellar
solution. The composition of the microemulsions used was chosen to correspond to the
monophasic area of the pseudoternary phase diagrams of the system determined at 25° C [4].
Traces of water present in olive oil and lecithin were determined by Karl Fischer titration and
taken into consideration in the calculation of the total water content of the system.

RESULTS & DISCUSSION

Four component microemulsions consisting of VOO or refined olive oil (ROO) as non-polar
solvent, lecithin as surfactant, propanol as cosurfactant and water were prepared. The choice of

11th International Congress on Engineering and Food - Athens, Greece, 2011 2127
the compositions of the microemulsions used was based on the pseudo-ternary phase diagrams
of the four-component system determined at 25° C for different weight ratios of the
components [4]. Alternative formulations using limonene and a series of biocompatible
alcohols were also tested. The structure of VOO microemulsions was studied by means of
various techniques, such as Electron Paramagnetic Resonance, Fluorescence Quenching,
Dynamic Light Scattering, Differential Scanning Calorimetry and Conductivity.

Figure 1. Pseudo-ternary phase diagrams for the four-component systems olive

oil/lecithin/propanol/water containing extra virgin olive oil (EVOO).The phase separation lines
correspond to the initially fixed weight ratio of lecithin/propanol: 2:1 (…) and 1:3 („). Compositions are
in weight ratios. The temperature was 25qC. [4]

It was found that the water molecules of the dispersed phase are bound to the amphiphiles.
When VOO was used, the size of the droplets was significantly smaller than the ones of ROO
due to the presence of the endogenous amphiphiles [5]. At water content above 3% (w/w) and a
high lecithin/propanol weight ratio a sharp increase in conductivity was observed, indicating a
structural transition in the bicontinuous form. The structure of the interface was influenced by
the concentration of the various components of the microemulsion system.

CONCLUSION
Microemulsions, as a food delivery system, may be a potential candidate to improve the
solubility and increase the bioavailability of food derived bioactive compounds.

REFERENCES
[1] Boskou D. Olive Oil: Chemistry and Technology; AOCS Press, Champaign IL USA, 1996.
[2] Georgalaki M.D. et.al J. Am. Oil Chem. Soc. 1998, 75, 155-159.
[3] Xenakis A. et.al 2010 Curr.Opinion Colloid Interface Sci. 15, 55-60
[4] Papadimitriou V. et.al Langmuir 2007, 23, 2071-2077
[5] Papadimitriou V. et.al J. Am. Oil Chem. Soc. 2005, 82, 335-340.

2128
Processing and technological characterization of extruded breakfast cereal obtained with

a mix of broken rice and common bean flour

Carvalho, A.V.a, Bassinello P.Z.b, Rios, A. de O.c

a
Embrapa Eastern Amazon, Belém, Brazil (anavania@cpatu.embrapa.br)
b
Embrapa Rice and Beans, Goiânia, Brazil (pzbassin@cnpaf.embrapa.br)
c
Federal University of Rio Grande do Sul, Porto Alegre, Brazil (alessandro.rios@ufrgs.br)

INTRODUCTION
Considering that the lack of good quality protein and calories in a diet can result in a
generalized malnutrition, then there is the possibility of exploiting the rice and beans through
the processing by extrusion cooking, with the possibility of obtaining products with good
quality technological, nutritional and sensory. The aim of this work was to evaluate the
functional technological properties of breakfast cereal obtained with a mix of broken rice and
common bean flour, by the analyses of expansion index, water absorption index, water
solubility index, apparent density, viscosity and instrumental texture.

MATERIALS & METHODS

The breakfast cereal was elaborated with a rice and common bean mixed flour, using the
proportion of 70% broken rice flour and 30% of broken common bean flour. The tested
formulation was processed in a single-screw extruder. The extrusion parameters were set using
three extrusion zones with temperatures of 40°C (1st zone), 60°C (2nd zone) e 80°C (3rd zone);
mixture moisture during processing set at 14%; screw speed set at 177 rpm; feeding rate of
290g/minute and circular matrix of 3.85mm. The developed breakfast cereal was sprinkled
with a sufficient quantity of a 70 ºBrix sucrose solution to make a final product with
approximately 35% sugar. The processed formulation was manually collected, submitted to a
forced air circulation oven drying and, afterwards, it was stored in polyethylene bags until
analyses. In order to characterize some technological performance of the breakfast cereal the
following analyses were performed: apparent density [1], radial expansion index [2], water
absorption and water solubility indexes [3], pasta viscosity (RVA), and instrumental texture
(texture analyzer Stable Micro Systems model TA.XT Plus).

RESULTS & DISCUSSION

The results for the technological characterization of the breakfast cereal of rice and beans are
presented on Table 1.

Table 1. Results of means and standard deviation for apparent density, expansion index and
instrumental texture of the breakfast cereal of rice and beans.
Apparent Expansion Water Water Texture (g.f)
density index absorption solubility index
index (g/g) (%)
0.25±0.00 8.89±0.14 6.41±0.13 44.50±0.14 1,087.44±220.44

11th International Congress on Engineering and Food - Athens, Greece, 2011 2129
The breakfast cereal of broken rice and common beans showed an apparent density of 0.25
similar to the values related by Ascheri et al. [4], who studied expanded products made of
whole amaranths and broken rice flours, and depending on the process conditions, reported
values varying from 0.13 to 0.68. The expansion index is a measure that allows one, at first, to
predict how severe or light the extrusion process was. The rice and beans breakfast cereal
showed an expansion index of 8.89, a value superior to those related by Vernaza et al. [5] for
breakfast cereal formulated with passion fruit bran. The water absorption (WAI) and water
solubility (WSI) indexes are parameters that help to measure the transformation degree
suffered by the amylaceous fraction of the extruded materials. The rice and beans breakfast
cereal had a WAI of 6.41g/g and WSI of 44.50% (Table 1). The texture is a critical factor for
the quality of crispy and expanded products. The rice and beans breakfast cereal had a hardness
value of 1087.44g.f. On the other hand, Vernaza et al. [5] studied different process conditions
to elaborate breakfast cereal of passion fruit bran and corn flour and found a wide range for
hardness, from 954g.f to 2623.73g.f The paste viscosity is an important attribute to study the
functional properties of starchy food, being one of the ways to evaluate the degradation degree
that occurs in these materials during thermal treatment. Severe treatments destroy the starch
granular structure, reducing the paste viscosity, what can be observed in this work through
viscosity values below 130cP.

CONCLUSION
It is possible produce extruded breakfast cereal using the proportion of 70% broken rice flour
and 30% of broken common bean flour and the final product shows good technological
properties.

REFERENCES
[1] Ramirez, J.L.A.; Wanderley, C.P. 1997. Effect de los parametros de extrusion, caracteristicas de pasta
y textura de pellets (snacks de terceira generacion) producidos a partir de trigo y maiz. Alimentaria,
279(1), 93-98.
[2] Alvarez-Martinez, L.; Kondury, K.P.; Harper, J.M. 1988. A general-model for expansion of extruded
products. Journal of Food Science, 53, 609-615.
[3] Anderson, R.A.; Conway, H.F.; Pfeifer, V.F.; Griffin, L. 1969. Gelatinization of corn grits by roll and
extrusuion cooking. Cereal Science Today, 14(1), 4-11.
[4] Ascheri, J. L.; Mendonça, X. M. F. D.; Ascheri, D. P. R.; Maia, M. C. A. 2005. Extrusão de harina
mixta de amranto integral y arroz: Parte I. Caracterización físico-química. Alimentaria, 367, 74-83.
[5] Vernaza, M.G.; Chang, Y.K.; Steel, C.J. , 2009. Efeito do teor de farelo de maracujá e da umidade e
temperatura de extrusão no desenvolvimento de cereal matinal funcional orgânico. Brazilian Journal
of Food Technology, 12(2), 145-154.

2130
 Cereal bar development using exotic fruit

Edmilson Rabelo Torresa, Everton Santana Castroa, Roneval Felix de Santanab, Juliana Cordeiro
Cardosoa,b, Cleide Mara Faria Soaresa,b, Álvaro Silva Limaa,b,*

a
Universidade Tiradentes, Aracaju-Sergipe, Brasil
b
Instituto de Tecnologia e Pesquisa, Aracaju-Sergipe, Brasil, *E-mail: alvaro_lima@unit.br

INTRODUCTION
Nowadays, the consumption of fast-foods and snacks has increased, but the desire for healthy
and functional foods increased at the same rate. In this sense the development of cereal bars
formulation presents itself as an emerging force for this niche market. Cereal bars are a popular
and convenient food and, therefore, would be an ideal food format to deliver fruit-derived
phenolic antioxidants and fibre [1]. Some ingredients can be used for the formulation of cereal
bars, including exotic fruits and organic residues are still in the enrichment of this product.
Papers report the use of cashew nut, passion fruit and amaranth. The goal of this research is
obtained cereal bars using exotic fruit (jenipapo and jack fruit seeds) in different composition
and compare sensorial and physic-chemically the cereal bars developed with standard and
commercial formulations.

MATERIALS & METHODS

Materials: The jenipapo pulp, jackfruit seed and all the raw materials to produce the cereal
bars was purchased of local market from Aracaju-Sergipe, Brazil. The chemicals were acquired
from Labsynth (Brazil) and had analytical grade. Cereal bar formulation: The cereal bars was
formulated using dry raw material (bran of oat -35% and rice – 15%) and agglutinative
ingredients (glucose syrup – 25% and honey – 25%). The exotic fruit was added to the base
formulation at different incorporation percentages. The agglutinative ingredients were heated
to 95oC and added to the dry ingredients, and then the bars were moulded (9x3x1, 5cm) and
cooled to -20oC for 10 min, packaged in aluminium and stored in a cool dry place at 25oC.
Centesimal composition: The cereal bars were evaluated for their physical-chemistry
characteristics, in triplicate (n=3) according to AOAC [2].Texture analysis: After 4 days of
storage, the hardness and crispness were determined using a texture analyzer. Sensorial
evaluation: A panel of 60 consumers was instructed to evaluate the sensorial attributes such as
appearance, taste, flavor, texture and overall linking. A 9-point hedonic scale with 1= dislike
extremely, 5= neither like nor like dislike and 9= like extremely was used. Statistical analysis:
Sensory data were analyzed using the Statistical Analysis System (SAS).

RESULTS & DISCUSSION

The cereal bars were formulated with jenipapo and jack fruit seeds in different concentration
(5, 10 and 15%) and evaluated by sensorial analyze of acceptability. The notes of attributes
sensorial to cereal bars decreased with increasing the incorporation of jenipapo from
approximately 7.0 (like moderately) to 5 (neither liked nor disliked). Statistical analysis
showed that the cereal bars with 5% of jenipapo were different from those containing 10% and
15%, which were statistically similar (p ” 0.05) among themselves. It was also observed that
all cereal bars formulated with jack fruit seed did not differ significantly (p < 0.05) for all

11th International Congress on Engineering and Food - Athens, Greece, 2011 2131
sensory parameters analyzed. In most cases, the notes for the sensory attributes were
approximately 7.0. Thus, the formulations with 15% of jack fruit seeds and 5% of jenipapo
were chosen to compare the sensorial, physico-chemical and texture characteristics among
cereal bars standard, commercial and formulated with exotic fruits. The cereal bar standard and
those formulated with jenipapo and jack fruit seed were extremely similar, the appearance
attribute did not significant difference (p < 0.05) for all cereal bars. In general, the cereal bars
had similar acceptance, with sensory note above 6.17. Freitas and Moretti [3] obtained average
scores for sensory evaluation of cereal bars banana flavor below of all score observed in this
work. The hardness values ranged between 942.01 (g) and 1900.04 (g), but the values did not
statistically different (p ” 0.05) among themselves. For the crispness attribute the change was
156.27 (g.s-1) 456.58 (g.s-1). The bars with jenipapo 5% and standard were similar when
compared with commercial cereal bars. Comparing the sensory analysis for attribute texture
and the instrumental value for this parameter, we can see that the cereal bar namely
commercial 1 had the highest score for texture, but the value for hardness and crispness were
the lowest values. As conclusion, the values of hardness and crispness influence in the
acceptance of the cereal bars. It is verified that the content total fiber of cereal bars formulated
with jenipapo (5%) and jack fruit seeds (15%) were 19.2 and 25.5 times, respectively, higher
than the cereal bar standard, and above 13.8 and 6.0 times when compare with cereal bars
commercial 1 and 2, respectively. Therefore, the cereal bars formulated in this work could be
considered as foods rich in fiber. The protein content was very low (approximately 0.05 for all
bars). The incorporation of exotic fruits allows smaller amounts of carbohydrates, a reduction
of approximately 14% of energy. Some authors report their research in the physical-chemical
values of cereal bars. Lima [4] using cashew to produce cereal bars, verified 7.40% of
moisture, 9.73% of protein 1.63% of ash, 9.70% of lipids and 5.84% of fiber, Freitas and
Moretti [3] produced cereal bars on the basis of textured soy protein and wheat germ, toasted,
with values of 10.71% moisture, 15.31% protein, ash 2 20%, lipid 5.64%, 5.17% fiber and
60.97 carbohydrates.

CONCLUSION
The highest incorporation of jenipapo and jackfruit seed in cereal bars, with greater sensory
acceptance, were 5% and 15% respectively. The comparison of the textures, physic-chemical
and sensory characteristics of the cereal bars formulated with exotic fruits and the standard and
commercial cereal bars indicate that the exotic raw materials can be use to formulated cereal
bars, besides the reduction approximately 14% of caloric value and increase the content fiber in
the cereal bar.

REFERENCES
[1] Sun-Waterhouse, D., Teoh, A., Massarotto, C., Wibisono, R. & Wadhwa, S. 2010. Comparative
analysis of fruit-based functional snack bars. Food Chemistry, 119, 1369–1379.
[2] AOAC. 1998. Official methods of analysis. Association of Official Analytical Chemistry, Arlington,
USA.
[3] Freitas. G.C. & Moretti, R.H. 2006. Caracterização e avaliação sensorial de barra de cereais funcional
de alto teor protéico e vitamínico. Ciência e Tecnologia de Alimentos, 26, 318-324.
[4] Lima, A.C. 2004. Estudo para a agregação de calor aos produtos de caju: elaboração de formulações
de frutas e castanha em barras. PhD Thesis. UNICAMP, Campinas- SP, Brazil.

2132
 Substitution of Ingredients by Green Coconut (Cocos nucifera L) Pulp in Ice Cream

Formulation

A. M. Igutia, A. C. I. Pereiraa, L. Fabianoa, R. A. F. Silvaa; E. P. Ribeiroa

a
Maua Institute of Technology, Sao Caetano do Sul, Brazil (amiguti@maua.br)

INTRODUCTION
Brazil is a great producer of coconut. Despite the volume of mature coconut consumed, an
increasing demand for green coconut has been observed to extract its water, that is a very
popular drink, consumed either in natura or after cartoon packaging. Beaches are places where
coconut water is consumed mainly in natura. As each fruit generates about 1.5 kg of residue, a
very serious environmental problem has to be solved because, according to estimative, of all
the garbage collected from the beaches, about 70% comes from green coconut. Two aspects are
more concerning: the big volume and the slow degradation [1]. Although many green coconut
water and husk researches have been developed lately, only a few about green coconut pulp are
found in scientific publications. This study intended to demonstrate that green coconut pulp can
replace milk, gums and emulsifiers in ice cream-like product formulation.

MATERIALS & METHODS

Green coconut from Paraiba State, Brazil, was purchased at Ceasa (Supply Centre of Santo
Andre, Sao Paulo). Fruits had the water extracted and the pulp separated. It was homogenized
and stored at -18 qC. To the process, liquid ingredients and the pulp were stirred and heated to
45-50 ºC. The powdered ingredients were added. The mixture was pasteurized at 87 °C for two
minutes and, after cooling, the ageing lasted 24 hours. Whipping and freezing were done in a
shaved surface heat exchanger. The product was kept at -18 ºC. Moisture, ash, sugar and
protein content were determined in triplicate according to methods described in AOAC. Lipid
content was determined by Bligh Dyer method. Overrun was determined by the % decrease in
weight of the product compared to the weight of same volume of mix used to produce it.
Melting time was determined by measuring the time to thaw 10 g of the product at 25 qC.
Acceptance testing was used to measure magnitude of like/dislike for final formulation, based
on a 9 point hedonic scale for overall liking. The seven different formulations tested included
the following ingredients, used all together or not: coconut pulp, cocoa powder, sucrose, water,
carrageenan gum, guar gum and hydrogenated vegetable fat. The final formulation had cocoa
powder (11%), green coconut pulp (41%), sucrose (17%) and water (31%). It is called sorbet.

RESULTS & DISCUSSION

Green coconut pulp is a very smooth material that changes according to maturation stage. The
more mature the fruit, harder and coarser is its pulp. In early development stages, the pulp is
jellylike; when completely mature, it becomes thicker, fibrous and hard, as reported in
literature [2]. The proximate composition of green coconut pulp also presents broad variation,
mainly because composition changes during nut development [2]. Most of its composition is
water. In dry basis, pulp used in this work presented 4.7% protein, 14% lipid, 4.1% ash and

11th International Congress on Engineering and Food - Athens, Greece, 2011 2133
77.1% carbohydrates. After process, the sorbet composition was 65.0% moisture, 2.4% protein,
1.0% lipid, 0.36% ash and 31.2% carbohydrate. It is lactose and cholesterol free and low fat
product. The product, according to Brazilian law, cannot be called ice cream because it does
not contain milk or solids from milk. It would be classified as sorbet because it contains more
than 20% total solids, and is prepared with fruit pulp.
The overrun result was 19.9%, much lower than values of overrun in industrial scale ice cream
production, which can be partially explained by the equipment. It did not permit overrun or air
cell size control, which are important parameters to ice cream quality [3].
In spite of low overrun results, the sorbet presented good creaminess, got from green coconut
pulp. It presented good sensory evaluation too. For this analysis, panellists were told that a
chocolate ice cream-like product was being evaluated. Almost 90% of the responses were “like
extremely” or “like very much”, which is a good evaluation of the product. About 93% of the
panellists would buy the product.
Ice cream is a system that contains air bubbles distributed in a frozen mix of water, fat, protein,
sweeteners, stabilizers and emulsifiers. Each component has its function in the system and,
besides its presence and proportions, the distribution, state of crystallization, droplet size and
spatial arrangement are critical for structure development of final product. That is why process
conditions are important. Studies have shown the importance of fat and emulsifier in ice cream
characteristics [4,5]. As the sorbet final formulation had no fat, stabilizers or emulsifiers added,
the good characteristics of the product should be attributed to coconut pulp and cacao
components. However, in our laboratory, we could verify that an ice cream-like product could
be formulated without cacao powder and that green coconut pulp has emulsifying and foaming
capacity (data not shown). To continue this research, it is necessary to find the ingredient
proportions to get the best results and verify how the development stage affects the product.

CONCLUSION
Components in green coconut pulp, probably proteins and carbohydrates, have emulsifier and
stabilizer properties, respectively. These properties permit development of cholesterol and
lactose free, low fat ice cream-like deserts.

REFERENCES
[1] Empresa Brasileira de Pesquisa Agropecuária. Embrapa Agroindústria Tropical. Aproveitamento da
Casca de Coco Verde. Available at:
http://hotsites.sct.embrapa.br/diacampo/programacao/2005/aproveitamento-da-casca-de-coco-verde.
[2] Grimwood, B.G. 1975. Coconut Palm Products – Their Processing In Developing Countries. Food and
Agriculture Organization of the United Nations. Fao. Rome.
[3] Sofjan, R.P. & Hartel, R.W. 2004. Effects Of Overrun On Structural And Physical Characteristics Of
Ice Cream. International Dairy Journal, 14, 255–262.
[4] Aime, D.B.; Arntfield, S.D.; Malcolmson, L.J. & Ryland. D. 2001. Textural Analysis Of Fat Reduced
Vanilla Ice Cream Products. Food Research International 34(2-3), 237-246.
[5] Segall, K.I. & Goff, H.D. 2002 A Modified Ice Cream Processing Routine That Promotes Fat
Destabilization In The Absence Of Added Emulsifier. International Dairy Journal 12, 1013–1018.

2134
 Evaluation of Drying Green Coconut Pulp for Obtaining a Snack-Like Product

W. H. Prieto; E. A. G. Seravalli; A. M. Iguti; M. Nitz

Mauá Institute of Technology, São Caetano do Sul, Brazil (wh.prieto@terra.com.br)

INTRODUCTION
The consumption of fresh or industrialized coconut water (Cocos nucifera L) in Brazil has been
increasing. A serious environmental problem has come up due to the inappropriate disposal of
the husks. It is estimated that 350 million liters of green coconut water are consumed in Brazil
every year and this consumption produces approximately 2 million tons of green coconut husk.
Barroso (2005) [1] showed that 70% of the waste generated in Brazilian coastal urban centers
is green coconut husk. Studies done in our laboratories show that the pulp rejected with the
husk presents interesting properties and can be used as an ingredient with potential applications
in food formulations. The objective of this study was to evaluate the feasibility of obtaining a
snack-like product by means of drying green coconut pulp. Drying will be performed in both a
cabinet dryer and a pulsed fluid bed dryer, assessing the product quality parameters and
relating them to the process variables.

MATERIAL & METHODS

Drying kinetics under different temperatures (60 °C, 70 °C and 80 °C) was determined with a
cabinet dryer produced by Armfield, UK. It consists of a 28 cm × 28 cm square duct and a fan
with adjustable speed, which blows the air through electric resistances, controlled by a PID
controller. The four-section pulsed fluid bed dryer used in this work has a total cross sectional
area of 0.18 m² (0.30 m × 0.60 m). Besides, the following characteristics were determined in
the raw and dried material: enzymatic activity (Polyphenol oxidase and peroxidase - method
described by CAMPOS & SILVEIRA (2003) [2]), water activity (was measured with a
Hygrometer (Aqualab Decagon Devices, Model Serie 3TE) at 25 °C)), moisture content (was
determined as described in A.O.A.C (1984) [3].), texture (analyses were performed using a
Texture Analyzer (TA- TA-XT2i, Stable Microsystems Ltd., UK) and color (parameter L -
were performed using the colorimeter COLOR EYE XTH).

RESULTS & DISCUSSION

In both dryers, drying was faster at 80 °C although there is no significant difference between
the profiles at 80 and 70 ºC. At 60 ºC, the drying rate is considerably lower, especially in the
cabinet dryer. The drying time required for both processes to reach equilibrium moisture
content was approximately 480 minutes in all tests. In Table 1 the initial and final moisture
contents, luminosity and water activity are presented. The temperature impacted the
equilibrium moisture content. Higher equilibrium values were obtained at lower temperatures,
which was expected for the equilibrium moisture content depends on the relative humidity. The
variations are due to the changes in ambient conditions. The variation of the water activity
values are explained by the different equilibrium moisture contents and the material lack of
uniformity. Values below 0.54 were obtained in all drying conditions. One can see clearly that
the parameter L*F decreased with the increase of temperature. An important consideration

11th International Congress on Engineering and Food - Athens, Greece, 2011 2135
 regarding the browning is that this is not caused by peroxidase and poliphenoloxidase content,
since these were not found in sufficient quantities to cause changes in color. Changes in the
color of coconut pulp are probably caused by the Maillard reaction, because the color, taste and
odor presented in the final product are typical of this reaction. In cabinet dryer to final
crispness (Force 1) was 5 ± 1 N and in PFB (Force 1) was 2 ± 1 N.

Table 1. Quality parameters in cabinet and PFB dryer.

T(°C) X0(db) Xe(db) Aw0 AwF L*0 L*F
Cabinet Dryer

60 (6.7 ± 0.4)a (0.277 ± 0.02)a (0.991 ± 0.001)a (0.38 ± 0.02)a (79 ± 2)a (72 ± 2)a
70 (6.5 ± 0.1)a (0.159 ± 0.004)b (0.993 ± 0.001)a (0.353 ± 0.002)b (83 ± 2)a (67 ± 1)b
a b a c a
80 (6.5 ± 0.1) (0.160 ± 0.003) (0.994 ± 0.002) (0.47 ± 0.02) (80.9 ± 0.7) (65 ± 2)b
T(°C) X0(db) Xe(db) Aw0 AwF L*0 L*F
a a a a a
60 (3.4 ± 0.4) (0.07 ± 0.01) (0.997 ± 0.001) (0.54 ± 0.04) (83 ± 3) (71 ± 1)a
PFB

a b a b a
70 (3.19 ± 0.08) (0.040 ± 0.004) (0.994 ± 0.001) (0.42 ± 0.01) (85 ± 1) (73 ± 3)a
80 (3.0 ± 0.5)a (0.036 ± 0.002)b (0.995 ± 0.001)a (0.42 ± 0.02)b (84 ± 2)a (63 ± 2)b
Means followed by same letters in column did not differ (p>0.05) according to the Duncan’s test.

CONCLUSION
Of the three process temperatures used (60, 70 and 80 ° C) that led to a more appreciated
product was 70 °C in 480 minutes of drying. Both process used resulted in a final product with
very similar characteristics to a snack, but due to the high fat content and lack of sample
homogeneity, only a few final pieces of the pulp showed the texture characteristic of chips. For
a industrial production of snack that would be require a study of green coconut pulp
maturation for chose the best maturation stage, because, in general, the higher the degree of
ripeness of the coconut, higher the fat content [4] and this directly influences the quality of the
final snack. For one low production, the results showed that the cabinet dryer is sufficient for
the manufacture of snack, since every time the tray contents are homogenized to prevent the
pulp sticking to the surface of the tray. In case of high demand, the indicated equipment is PFB
dryer because it has great productivity by allowing the drying of large amounts of green
coconut pulp.

REFERENCES
[1] Barroso, T. Fortaleza Ganha primeira Unidade de Beneficiamento de Casca de Coco Verde do
Nordeste. Início. Imprensa. Notícias. 30 jun. 2005. Disponível em: <http://www.embrapa.br>. Acesso
em: 19 mar. 2008.
[2] CAMPOS, A. D.; SILVEIRA, E. M. da L. Metodologia para Determinação de Peroxidase e da
Polifenoloxidase em Plantas. Comunicado Técnico EMBRAPA, 87, Pelotas, RS, p. 1-3, Abril, 2003.
[3] A. O. A. C. (Association of Official Analytical Chemists). Official Methods of Analysis. 14. ed.
Arlington: A.O.A.C., 1984. 1141p.
[4] Aragão, W. M.; Cruz, E. M. de O.; Tavares, M.; Ribeiro, F. E.; Tupinambá, E. de A.; Pimentel, S. A.;
Takemoto, E. Teor de Gordura e Composição de Ácidos Graxos em Polpa de Frutos de Coqueiro
Anão em Diferentes Idades de Maturação. Revista do Instituto Adolfo Lutz. São Paulo, v. 63, n. 2, p.
159-167, 2004.

2136
 Physical-Chemistry and Microbiological Analysis of Probiotic Dairy Beverage

Fermented with Kefir

Luiz Rodrigo Ito Morioka1; Maria de Fátima Fonseca1; Luciano Avallone Bueno2*; Djalma Marques1;
Graciliane Cruz Ximenes3; Cynthia Souza1; Marcos Antônio de Morais Jr 3

1
BioLogicus – Research Center of Probiotics, Technology Institute of Pernambuco, Recife, Pernambuco,
Brazil;(www.biologicus.com.br)
2
University Rural of Pernambuco, Physical Department, Recife, Pernambuco, Brazil;
*(bueno@biologicus.com)
3
University of Pernambuco, Recife, Pernambuco, Brazil

INTRODUCTION
The concern with respect to food has been changing in recent decades, thus the concept of
functional foods made science such as nutrition and medicine can be associated, earning extra
dimension in the XXI century. Probiotics are described as live microorganisms which when
administered in adequate amounts confer benefits to its consumers. The incorporation of
probiotics as dietary adjuncts in different dairy products has strengthened its functional
properties, resulting in increased consumption, along with excellent sensory characteristics.
The cheese whey, called sweet (pH between 6 and 7), besides being a secondary product of the
cheese factory industry, is resulting from the enzymatic coagulation of milk. This secondary
product in addition to representing approximately 85 - 95% of the initial volume of milk used
to manufacture cheese it contains approximately 55% of the nutrients in milk. The conversion
of liquid whey in dairy beverages fermented or not, it would be one of the most attractive
options for the industries due to the simplicity of process, the possibility of using the
equipment already in the milk processing; the replacing the use of whey powder, reducing
costs; besides the reduction of problems relating to their disposal. The aim of this study was to
evaluate the physical-chemical characteristics and microbiological characteristics of fermented
dairy beverages made with 50% of pasteurized milk and 50% of whey added to the culture of
kefir.

MATERIALS & METHODS

During fermented dairy beverages it was used whey from the production of minas fresh cheese
and pasteurized milk from Faco´s Dairy, located in Ribeirão-PE. Probiotic culture utilized was
obtained by kefir milk. Reagents used in the physical-chemistry analysis were all analytical
grade. Preparation of dairy beverages, obtained from milk and cheese whey, was based on
methodology with modifications. Mixture of milk and whey (1:1) supplemented with sucrose
(10g/100g mixture of milk and whey), Estabgem 83 (0.45%), modified starch (1.3%) and
sorbate (0.03%) was pasteurized at 74°C for 15 minutes. Fermentation was conducted until pH
around 4.6, and stored at 5 ± 1ºC for overnight. Finally, there was clot breaking and after that
natural pulp, natural coloring and natural strawberry flavour were added. Dairy beverages were
assessed for lactic acid bacteria viability (CFU/g), Salmonella spp/25g, coliforms at 45°C
(MPN/g) and yeasts (CFU/g) [1, 2]. After inoculations, dairy beverages were incubated for

11th International Congress on Engineering and Food - Athens, Greece, 2011 2137
fermentation at 42°C until pH 4.6 was reached. The beginning of fermentation was represented
by time 0 min and control fermentation process was monitored by analyzing pH measure in a
digital potentiometer [3] and acidity levels were carried out by titration alkalimetric acid, using
phenolphthalein as indicator. When pH of dairy beverage approached 4.6 the fermentation was
stopped by cooling and stored under refrigeration (4°C) for 22 days. During fermentation
samples were collected for physical-chemistry analysis.

RESULTS & DISCUSSION

Presumptive test for coliforms at 45°C and Salmonella spp proved negative for presence of the
same. Due to the low pH of the product it is known that these microorganisms can suffer stress
and not be detected in the analysis. Lower amount of coliforms at 45°C expressed as < 3.0
NMP/g represents no growth, considering limit of the method. Thus, the absence of coliforms
at 45°C in the final product may also be indicative of good hygienic and sanitary conditions,
during the preparation of dairy beverage. However, it is not detected the presence of yeasts in
beverages analysed. The result for the lactic acid bacteria was within the standard scores based
on the Normative Instruction n° 36, 21/12/2000 (Ministry of Agriculture) and RDC Resolution
n° 12[4], 02/01/2001 (ANVISA)[5] establishing the minimum 106 CFU/mL of lactic acid
bacteria in fermented dairy beverages. The stability, aroma, flavor and texture of fermented
dairy products depend on the pH. The results obtained from the fermentation of dairy beverage,
showed a gradual decrease in pH and acidity of the product. Decrease in pH during
fermentation can be attributed to symbiotic relationship between microorganisms present and
in the culture probiotic kefir. The associated growth of these microorganisms results in shorter
clotting fermentation, increased production of lactic acid and increased stability, aroma, flavor
and texture in final product.

CONCLUSION
Dairy beverage analyzed attends item 8Fb RDC 12/2001 - ANVISA, regarding the parameters
required for the sample indicative. Thus, showing good hygienic and sanitary conditions, since
not detected coliform at 45°C and Salmonella spp. Study of fermentation cultures for probiotic
dairy drink kefir had good results with respect to fermentation time of only 4.5 hours. Total
number lactic acid bacteria in fermented beverage stored at 5°C remained above 107 CFU/g
during 22 days, values that are consistent with the required probiotic products legislation.

REFERENCES
[1] American Public Health Association. 2001. Compendium of Methods for the Microbiological
Examination of Foods. 4ª ed. APHA.
[2] Association of Official Analytical Chemists, 17th Edition, 2002. AOAC (967.26); AOAC (966.24);
AOAC (997.02).
[3] Instituto Adolfo Lutz. 2005. Normas Analíticas do Instituto Adolfo Lutz. 4º ed. Brasília.
[4] Instrução Normativa N.º 36 de 31 de outubro de 2000. Aprova o Regulamento Técnico de Identidade
e Qualidade de Bebidas Lácteas. http://extranet.agricult
[5] Brasil, Agência Nacional de Vigilância Sanitária RDC nº. 12, de 02 de janeiro de 2001. Regulamento
Técnico sobre os Padrões Microbiológicos para Alimentos. Disponível em: http://anvisa.gov.br/legis

2138
 Phytochemicals and antioxidant activity of comminuted orange (Citrus sinensis L.)

Zamantha Escobedo-Avellanedaa, Vinicio Serment-Morenoa, Aurora Valdez-Fragosoa, Hugo Mujica-

Paza, and Jorge Welti-Chanesa

a
Department of Biotechnology and Food Engineering, ITESM, Monterrey, Nuevo León, Mexico
(zamantha.avella@gmail.com, vsermentm@gmail.com, a.valdez@itesm.mx, hmujicap@yahoo.com,
jwelti@itesm.mx)

INTRODUCTION
Orange is an important source of phytochemicals that due to an antioxidant mechanism can
reduce the incidence of chronic illness. Both juice and peel contain phytochemicals, but they
are more abundant in peel. “Comminuted” is a product obtained by grinding peel and orange
juice, and mainly used as a natural basis to enhance sensory properties of soft drinks.
Phytochemicals and antioxidant activity of comminuted must be evaluated in order to
determine the level in which this product is a source of these compounds and can provide
antioxidant activity. Thus, the aim of this work was to quantify phenolic compounds, vitamin
C, total carotenoids, and antioxidant activity of comminuted orange.

MATERIALS & METHODS

Comminuted orange preparation: Juice, flavedo and albedo (71.6, 16.0 and 12.4% w/w
respectively) of commercially mature Valencia oranges were grinded to obtain a homogenous
paste. Phenolic compounds: 250 mg of comminuted were mixed with either 5 mL of
methanol: water (1:1 v/v) (for free phenolics (FP)) or 1.2 M HCl in (1:1 v/v) methanol/water
(for total phenolics (TP)). The mixture was heated at 90 °C/ 3 h and then centrifuged. 50 PL FP
or TP extracts were mixed with 650 PL of water and 50 PL of the Folin-Ciocalteau reagent.
After 5 min incubation at room temperature, 250 ȝL of 1 N Na2CO3 were added and again
incubated (37 °C/2 h). Absorbance was read at 765 nm. Vitamin C: 0.25 g of comminuted and
1 ml of 6% TCA were centrifuged. 25 PL of 75 mM potassium phosphate buffer and 50 PL of
supernatant were added to microcentrifuge tubes for both reduced (RA) and total ascorbate
(TA). 25 Pl of 10 mM DTT were added to the TA tubes and then incubated at room
temperature/10 min. 25 Pl of 0.5% NEM were added to the same tubes and 50 Pl of water to
the RA tubes. 125 Pl 10% TCA, 100 Pl 43% H3PO4, 100 Pl 4% DD´-bipyridyl and 50 Pl 3%
FeCl3 were added to all tubes and then incubated at 37 °C/1 h. Absorbance was read at 525
nm. Total carotenoids: 75 mg of comminuted were mixed with 1.5 ml of ethanol: THF (1:1
v/v). An ultrasound treatment at 45 W/10 min was applied. Samples were centrifuged and
absorbance was read at 450 nm. Antioxidant activity: 25 Pl of adequate dilution of
comminuted was placed in black plates with 96 wells. A microplate reader was used to
automatically dispense 150 Pl of 1 PM fluorescein. After 30 min of incubation/37 °C, 25 Pl of
153 M AAPH were dispensed. Fluorescence (485 nm ex and 528 nm em) was read at 37 °C.

RESULTS & DISCUSSION

Phytochemicals analyzed in this work are shown in Table 1. The amount of TP and FP was
found as 285.5±9.0 and 181.1±7.4 mg GAE/100 g respectively. FP were subtracted from TP,
and 36.6% of conjugated phenolics were calculated, indicating that most phenolics are in free

11th International Congress on Engineering and Food - Athens, Greece, 2011 2139
form. Concentration of TP in comminuted orange is similar to that reported by the USDA [1]
for navel orange (337 mg GAE/100 g wb). Comminuted orange could be considered as an
important source of phenolics when compared with foodstuffs widely recognized by their high
phenolic content, such as blackberry and blueberry (447 and 311 mg GAE/100 g wb
respectively) [1]. L-ascorbic acid (reduced form) and dehydroascorbic (oxidized form) account
for total vitamin C. 48.8±1.5 and 44.9±1.1 mg L-AAE/100 g for TA and RA were found
respectively for comminuted orange. Only 8% was found as dehydroascorbic acid. The USDA
[1] reported 50 and 136 mg L-AAE/100 g wb of total vitamin C for juice and peel orange
respectively. Total carotenoids in comminuted orange was 3.3±0.2 mg CE/100 g wb, while in
dry basis this value corresponds to 21.0±1.1 mg CE/100 g db. 8 mg CE/100 ml freeze dried
orange juice, and 44.5 mg CE/100 g freeze dried peel were reported by Yuan-Chuen et al. [2]
and Yuan-Chuen et al. [3] respectively. Using data from these authors, and considering the
orange juice and flavedo plus albedo (peel) proportions used in comminuted orange
formulation, the total carotenoid content calculated is 18.4 mg CE/100 g db, value similar to
21.0±1.1 mg CE/100 g db. Because phenolics are 86.5 times higher than total carotenoids, they
are the main phytochemicals in the product. Antioxidant capacity, mainly attributed to
hydrophilic compounds such as phenolics and vitamin C, was 3606.6±154.9 mol TE/100 g, this
value is much higher than that reported by the USDA [4] for orange juice (726 mol TE/100 g).
Compared with other foodstuffs widely recognized as sources of antioxidants such as
blackberry and blueberry with 5802 and 3463 mol TE/100 g respectively [4], the antioxidant
capacity of comminuted could be considered as high.

Table 1. Concentration (in wet basis) of some phytochemicals of comminuted orange.

Phenolics (mg GAE/100 g) Vitamin C (mg L-AAE/100 g) Carotenoids Antiox. activity
Total Free Total Reduced (mg-CE/100 g) mol TE/100 g)
285.5±9.0 181.1±7.4 48.8±1.5 44.9±1.1 3.3±0.2 3606.6±154.9
GAE, gallic acid equivalents; L-AA, L-ascorbic acid equivalents; CE, carotene equivalents; TE, trolox
equivalents.

CONCLUSION
Knowledge of the concentration of phytochemicals in comminuted orange is fundamental for
evaluating the level in which this product is a source of these compounds, and can provide
antioxidant activity. This is also important for designing the most adequate method to preserve
it without reducing phytochemicals availability and functionality. Results show that orange
comminuted can be recognized as an important source of phytochemicals like foodstuffs such
as blackberry and blueberry.

REFERENCES
[1] USDA National Nutrient Database for Standard Reference. www.nal.usda.gov/fnic/foodcomp
[2] Yuan-Chuen W., Yueh-Chueh C. & Yu-Hua K. 2007. Quantitation of Bioactive Compounds in Citrus Fruits
Cultivated in Taiwan. Food Chemistry, 102, 1163–1171.
[3] Yuan-Chuen W., Yueh-Chueh C. & Hsing-Wen H. 2008. The Flavonoid, Carotenoid and Pectin Content in Peels of
Citrus Cultivated in Taiwan. Food Chemistry, 106, 277–284.
[4] U.S. Department of Agriculture, Agricultural Research Service. 2010. Oxygen Radical Absorbance Capacity
(ORAC) of Selected Foods, Release 2. Nutrient Data Laboratory Home Page: www.ars.usda.gov/nutrientdata/orac

2140
11th International Congress on Engineering and Food - Athens, Greece, 2011 2141
2142
 Sensory and antioxidant properties of beer with Juniperus communis L.

Mile Veljovic1, Sasa Despotovic, Radovan Djordjevic, Sonja Pecic, Ana Kalusevic, Ida Leskosek-
Cukalovic, Viktor Nedovic
1
Faculty of Agriculture-University of Belgrade, Belgrade-Zemun, Serbia (mileveljovic@yahoo.com)

INTRODUCTION
Juniperus communis L. is a coniferous shrub with fruits described as berries or berry-like
cones. The berry or fruit of this species have a aromatic, spicy aroma, and a slightly bittersweet
flavour. Juniper berries have long been used for flavouring foods and beverages and as a
remedy for many health problems. The medieval pre-hops brewers used different herbs or
mixtures of herbs (called grut) to flavor their beers. The most common of these were sweet
gale, juniper, yarrow, rosemary, mugwort, and woodruff. In this study investigated were
sensory and antioxidant properties of beers produced with three different concentrations of
juniper berries. The goal was to determine the influence of adding of juniper berries on sensory
characteristic and antioxidant properties of the obtained beers.

MATERIALS & METHODS

Juniper berries were cut into two pieces and added to the wort in 3 different concentrations:
0.24, 0.48 and 0.72 g/L. The mixtures of wort and juniper berries were sterilized and seeded
aseptically. The fermentation process was completed after 25 days. The control beer was
obtained by fermentation of pure wort. The physico-chemical properties of the beers were
determined using Alcolyzer Beer ME Analyzing System, Anton Paar GmbH – AUSTRIA.
The total phenolic content of samples was determined by the Folin-Ciocalteu method [1]. The
FRAP assay was performed according to the procedure described by Benzie and Strain [2].
Aqueous solutions of known ascorbic acid concentrations were used for calibration. The
DPPH-reducing activity was estimated following the procedure described by Kaneda et al [3].
The TEAC assay was conducted using method described by Re at al [4]. A sensory evaluation
of the obtained products was conducted, assessing: fragrance, taste, aroma, body, bitterness,
freshness and general impression. A commercial lager beer was used as a reference for the
assessment. The experimental results were analyzed with the student’s t-test for dependent
samples.

RESULTS & DISCUSSION

The physico-chemical characteristics of the beers are shown in Table 1.

Table 1. Physico-chemical properties of beers

Parameter Cb K1 K2 K3
Original gravity (°Plato) 11.91 11.91 11.91 11.91
Real extract (% w/w) 2.80 3.17 3.80 3.66
Real degree of fermentation (% w/w) 77.60 74.00 69.69 70.28
Alcohol (% v/v) 5.00 4.37 4.23 4.18
Calories (kJ/100 mL) 148.00 174.60 180.69 177.02
Cb – control beer; K1, K2, K3 – beers with 0.24, 0.48 and 0.72 g/L of juniper berries, respectively

11th International Congress on Engineering and Food - Athens, Greece, 2011 2143
Phenolic compounds are generally considered as one of the very important antioxidant sources
in beer. Total phenolic content and the antioxidant capacity of beer samples are shown in Table
2. In the beer samples with juniper, the total amount of phenolic compounds was significantly
higher than in the control beer. However, mutual comparison of samples K1, K2 and K3 were
shown statistically significant difference only between samples K1 and K3.

Table 2. Total antioxidant capacity of beers

Assay
Sample a b
TPC (mg GAE /l) FRAP (mM AC) TEAC (mM TE)c DPPH (%)d
Cb 385,6 ± 6,6 3,61 ± 0,04 3,27 ± 0,04 50,59 ± 0,88
K1 416,2 ± 7,3 3,73 ± 0,07 3,71 ± 0,02 64,70 ± 1,27
K2 432,8 ± 5,4 3,84 ± 0,03 4,37 ± 0,02 69,00 ± 1,08
K3 450,0 ± 3,1 3,90 ± 0,03 4,45 ± 0,03 69,56 ± 1,40
Each value is the mean ± standard deviation of three replicate experiments. Bolded numbers indicate
significantly different values (p < 0,05) compared with control beer; Cb – control beer; K1, K2, K3 –
beers with 0.24, 0.48 and 0.72 g/L of juniper berries, respectively; a total phenolic content, expressed as
milligrams of gallic acid equivalents per liter of beer; b total antioxidant capacity expressed as mmol of
ascorbic acid equivalents; c total antioxidant capacity expressed as mmol of Trolox equivalents;d % of
inhibited of DPPH free radical after 30 minutes.

Using the TEAC and DPPH assays, it was found statistically significant difference between
control beer and juniper beers. However, the FRAP method was shown significant difference
only between control beer and sample K3. The results of the sensory evaluation are presented
in Figures 1. Sensory analysis suggests that beers produced with juniper berries have more than
satisfactory sensory properties, even better than a common lager beer.

Figure 1. The results of sensorial evaluation of beer samples

CONCLUSION
The obtained results suggest that beers produced with different proportion of juniper berries
have very interesting and pleasant sensory properties. The addition of juniper berries can give
very pleasant taste and flavor to beer but also increase its antioxidant capacity.

REFERENCES
[1] Singleton V.L. & Rossi J.A. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagent.
American Society for Enology and Viticulture, 16(3), 144-158.
[2] Benzie I.F.F. & Strain J.J. 1996. The ferric reducing ability of plasma (FRAP) as a measure of ”antioxidant power”:
The FRAP assay. Analytical Biochemistry, 239(1), 70-76.
[3] Kaneda H., Kobayashi N., Furusho S., Sahara H. & Koshino S. 1995. MBAA Technical Quarterly, 32(2), 90-94.
[4] Re R., Pellegrini N., Proteggente A., Pannala A., Yang M. & Rice-Evans C. 1999. Antioxidant activity applaying
an improved ABTS radical cation decolorization assay. Free Radical Biology & Medicine, 26(9-10), 1231-1237.

2144
 Influence of phytosterols addition in the rheology and sensory attributes of dark

chocolate

Priscilla Efraima; Gabriela C. Marsona; Denise C.P. Jardimb; Aline O. Garciab; Katumi Yotsuynagib

a
Universidade Estadual de Campinas, Campinas, Brazil (efraim@fea.unicamp..br,
gabrielacmarson@gmail.com)
b
Instituto de Tecnologia de Alimentos (ITAL), Campinas, Brazil (djardim@ital.sp.gov.br,
alinegarcia@ital.sp.gov.br, katumyot@ital.sp.gov.br)

INTRODUCTION
Chocolate is a food consumed and enjoyed worldwide by people of varied ages and social
classes. It is considered a non-Newtonian fluid, pseudoplastic, and its rheological behavior
determines the process conditions adopted industrially. Phytosterols are plant sterols found in
vegetables and have proven to be effective in reducing levels of total cholesterol and LDL-
cholesterol through inhibition of cholesterol absorption [1]. Studies about the application of
phytosterols in chocolate and its influence on sensory characteristics and technological
performance are scarce. Thus, this study evaluated the influence of phytosterols in dark
chocolate on rheological and sensory properties, in order to obtain products with suitable
technological and sensory levels, also providing health benefits.

MATERIALS & METHODS

From a basic formulation containing 55.0% of natural cocoa liquor, 39.9% sugar, 5.0% cocoa
butter deodorized and 0.1% of vanilla flavor were produced 6 samples of chocolate with 3
types of phytosterol and 6 samples of free-phytosterol chocolate (standard for comparisons), in
which the levels of the emulsifiers lecithin and polyglycerol poliricinoleate were varied from
0,0 to 0,5% (sum of both). It was used the following phytosterols in the following amounts
(corresponding to 0,8g of phytosterols/40g of chocolate): Encapsulated pine phytosterol
powder (A), 2.7 g/100g of chocolate; Oil-based soy phytosterol (B), 3.6 g/100g of chocolate;
Powder soy phytosterol (C), 2.1 g / 100g of chocolate.
The rheological determinations were performed according to Vissotto et al. [2] at 40°C. The
sensory evaluation consisted of an acceptability test with 60 consumers [3] of the global
acceptability, aroma, flavor, melting in the mouth and greasy feel in the mouth after
consumption by 9-point hedonic scale (9: extremely like and 1: extremely dislike); force
required to break the chocolate using an scale of the ideal of 7 points (7: requires much more
strength than I like, and 1: requires much less force than I like) and purchase intention (5:
definitely would buy and 1: definitely wouldn’t buy). For the statistical analysis, data
underwent ANOVA and Tukey test to compare means [4].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2145
RESULTS & DISCUSSION
Plastic viscosity (Pa.s) x Emulsifier content Chocolate yield value (Pa) x Emulsifier content

9 30
Standard
8 A
25
Plastic viscosity (Pa.s)

B
7
C
6 20
Standard
5 A
15
B
4
C
3 10

2
5
1

0 0
0,5 : 0 0,4 : 0,1 0,3 : 0,2 0,2 : 0,3 0,1 : 0,4 0,0 : 0,5 0,5 : 0 0,4 : 0,1 0,3 : 0,2 0,2 : 0,3 0,1 : 0,4 0,0 : 0,5

% Lecithin : % PGPR % Lecithin : % PGPR

(a) (b)
Figure 1. Casson plastic viscosity (a) and yield value (b) of the chocolates with and without phytosterols
after the conching step

The addition of the oil-based phytosterol (B) did not affect the sensory attributes of dark
chocolate and affected positively the plastic viscosity and the yield value, reducing their
values. On the other hand, the addition of the encapsulated phytosterol (A) affected the sensory
acceptability and influenced negatively the plastic viscosity and the yield value of the
chocolates. The addition of phytosterol in the powder form (C) also influenced negatively the
rheological parameters, but did not influence the sensory attributes.

CONCLUSION
It was demonstrated that the different type of phytosterols added to chocolate influenced its
sensory properties and its rheological behaviour in a different way, but the technological
feasibility of the application was also proved. This study will be improved assessing the shelf
life of the chocolates with the phytosterols and if there is any loss or oxidation of the
phytosterols during this period.

REFERENCES
[1] Cercaci, L.; et al. (2007). Phytosterol oxidation in oil-in-water emulsions and bulk oil. Food
Chemistry, 102,161-167.
[2] Vissotto, F. Z. et al. (1999). Caracterização físico-química e reológica de chocolates comerciais
elaborados com gorduras alternativas. Brazilian J. Food Techn., 2(1), p.139-148.
[3] Meilgaard, M.; Civille, G.V.; Carr. B.T. 1987. Sensory Evaluation Techniques. CRC Press. New
York. NY. 281p. [4] SAS Institute Inc. Sas/STAT User’s Guide. Release. Cary, NC: SAS Institute
Inc, 1028p. 1993.

2146
 Addressing new functional fillo products through nutrition and healthy ingredients: Hi

omega-3 fatty acids and phytosterol esters

Varzakas, T.a, Labropoulos, Ab, and Anestis, S.b

a
Technological Institute of Kalamata, Hellas (tvarzakas@teikal.gr)
b
Technological Institute of Athens, Hellas (athanlab@teiath.gr)

INTRODUCTION
Hi Flaxseed Oils and other microencapsulated Hi omega-3 marine oriented products are
excellent sources of Essential Fatty Acids (EFAs), including Omega-3 (alpha-linolenic acid),
Omega-6 (linoleic acid) and Omega-9 (oleic acid). Athens Foods has launched an omega–3
fortified fillo dough aimed at health and wellness oriented customers with the following health
claim as defined for foods containing any level of EPA and DHA which meet the qualifying
criteria. This study has been designed to incorporate new functional (fortified) foods into fillo
dough products. It reviews all the key issues addressed, key problems to be solved and methods
to achieve success.
The development process will be explained and more specifically how the functional
ingredients were chosen, how they were presented to the consumer, recipes incorporated,
claims made, branding made, product liability with unknown risks and how all these will be
communicated with the consumers. Two case studies will be presented.

MATERIALS & METHODS

Two formulas for omega-3 FA and plant sterols fillo doughs are given. Torsion test, Kramer
shear test and Texture profile analysis (TPA) were the methods used to determine texture.
Kramer shear test and TPA are empirical tests whereas Torsion test is a fundamental test for
texture according to Kim et al. [1]. Shear stress and shear strain of cooked gels (fillo dough
products) was determined using torsion test [2, 3].
Colour properties were determined using a Minolta Chroma Meter CR-400 colorimeter. A CIE
color system using L ,a b tristimulus color values was employed.
Score cards with five lines have been provided for sensory evaluation (appearance, odour,
texture, flavour and overall acceptability) for each sample.

RESULTS & DISCUSSION

Texture profile analysis (Table 1) revealed no differences (P>0.05) in the six texture
parameters of the cooked fillo dough products compared to fillo dough products with added
neutraceuticals. Similar TPA values have been reported by Kassis et al. [3]. Stabilization might
have occurred due to yolk phospholipids and soybean lecithin contained in omega-3 and
phytosterol esters.
Finally all products with or without neutraceuticals had similar Kramer shear force (P>0.05)
averaging approximately 140 N/g. However, it should be noted that fat emulsification results in
stabilization of food products which in turn requires more shear force to fracture a sample. This
is also confirmed by torsion shear stress and TPA hardness values. Finally there was no

11th International Congress on Engineering and Food - Athens, Greece, 2011 2147
difference (P>0.05) in shear strain between the tested samples ranging from 0.93 to 1.1
confirming the TPA cohesiveness values since shear strain is a measure of gel cohesiveness.

Table 1. Texture profile analysis of cooked fillo dough products with added omega-3 fatty acids and
phytosterol esters compared to fillo dough products with nothing added

Fillo dough Fillo dough with Fillo dough with

omega-3 FA phytosterol esters
Springiness 2.02 1.97 1.92
Hardness 1400 1405 1423
Cohesiveness 0.65 0.67 0.69
Gumminess 985 992 994
Chewiness 1520 1600 1585
Resilience 0.33 0.34 0.35

Figure 1. Different uses of

fillo dough

CONCLUSION
Functional food containing neutraceuticals is highly encouraged. However, attention needs to
be paid at the level of concentrations added since there is no Recommended Daily Allowance
and the concentrations in the tablets or capsules are not clear. However, questions arise from
consumers regarding credibility and price, new brands, product formats and health claims.

REFERENCES
[1] Kim, B.Y., Park, J.W., and Yoon, W.B. 2005. Rheology and texture properties of surimi gels. In J.W.
Park (ed.) Surimi and surimi seafood (2nd edition). Pp. 491-582, Boca Raton, FL: Taylor and Francis
Group.
[2] Taskaya, L., Chen, Y.C., Beamer, S., and Jaczynski, J. 2009A. Texture and colour properties of
proteins recovered from whole gutted silver carp (Hypophthalmichthys molitrix) using isoelectric
solublization/precipitation. Journal of the Science of Food and Agriculture, 89 (2), 349-358.
[3] Kassis, N., Drake, S.R., Beamer, S.K., Matak, K.E., Jaczynski, J. 2010. Development of
neutraceutical egg products with omega-3-rich oils. LWT-Food Science and Technology, 43, 777-
783.

2148
 The non–starch polysaccharides quantity changes in pastry products where Jerusalem
artichoke (Helianthus tuberosus L.) added

Ilga Gedrovicaa, Daina Karklinaa, Anna Frasb, Olga Jablonkab, Danuta Borosb

a
Faculty of Food Technology, Latvia University of Agriculture, Jelgava, Latvia (Ilga.Gedrovica@llu.lv)
b
Laboratory of Quality Evaluation of Plant Materials, Institute of Plant Breeding and Acclimatization,
Radzikow, Poland (postbox@ihar.edu.pl)

INTRODUCTION
In recent years people around the world and also in Latvia have become more and more
interested in healthy food. Foods containing dietary fiber (DF) constitute a major component of
a healthy, balanced diet. A good source of non-starch polysaccharides (NSP) fiber is vegetable
Jerusalem artichoke (Helianthus tuberosus L.) that may be useful for treatment of various
gastrointestinal disorders. Jerusalem artichoke (JA) consumption helps lowering cholesterol
levels, reducing the risk of colon cancer, and losing weight [1]. The addition of Jerusalem
artichoke powder (JAP) to pastry products is one more possibility of making a healthier food
and increasing DF intake, however, JAP concentration in products is limited from
technological and sensory aspects. More important is the consumer sensory evaluation, which
showed that people liked cakes, honey biscuits and butter biscuits with 30 % JAP and part of
the consumers agree to use pastry products with 50 % of JAP. The aim of the study was to
determine changes in quantity of individual NSP in raw JAP and such pastry products as cakes,
butter biscuits and honey biscuits with JAP added.

MATERIALS & METHODS

The experiments were carried out in the Laboratory of Quality Evaluation of Plant Materials at
the Institute of Plant Breeding and Acclimatization at Radzikow in Poland. Analyzed cakes,
honey biscuits and butter biscuits baked using classic recipe and technology, were used as a
control samples. In samples with JA part of wheat flour was substituted with JAP in
concentrations 30 % and 50 %. The Upsala method includes preparation of a residue after
treatment with thermostable D–amylase and amyloglucosidase and then ethanol precipitation of
solubilized DF components [2]. After acid hydrolysis of total residue in two steps, neutral
polysaccharide residues are quantified as alditol acetates by gas–liquid chromatography, uronic
acid residues determined by colorimetry, and the ash–free acid insoluble residue is determined
gravimetrically. With this method, total DF, including starch resistant to the enzymatic
treatment used, is calculated as the sum of analyzed polysaccharides and Klason lignin. Fructan
is not included in the analysis.

RESULTS & DISCUSSION

Results of this study showed that the major NSP monomers of JAP and pastry products are, in
decreasing manner, hexoses: glucose, galactose, mannose, and pentoses: arabinose, xylose,
rhamnose, and fucose. Total amount of NSP is 9.2 % in JAP; out of them 6.3 % composes of
hexoses – and 2.9 % of pentoses (Fig.1, A). International Life Sciences Institute (ILSI) declares
that to claim as “source of DF”, a food must contain at least 3 g of DF per 100 g for a solid
food [3]. The NSP amount in classical cake and butter biscuits is less than 3 g per 100 g, only

11th International Congress on Engineering and Food - Athens, Greece, 2011 2149
honey biscuits has higher level of NSP. Adding JAP to pastry products significantly increases
the amount of NSP (pd0.05). The concentration 30 % of JAP in total amount of flour increases
NSP amount in cakes by 52.3 %, in honey biscuits by 49.3 %, but in butter biscuits by 48.4 %,
also permits to say that these pastry products are “source of DF”. When JAP added in quantity
of 50 %, the NSP contents go up in cakes by 86.2 %, in honey biscuits by 68.9 %, but in butter
biscuits by 69.4 % as compared with control samples (Fig.1, B).
Total pentoses and hexoses 9.2 9
8 control 30 % of JAP 50 % of JAP

T o ta l a m o u n t o f N S P
Total hexoses 6.3
4.1 A 7 5.7 B

in g p er 1 0 0 g
hexose

glucose
galactose 1.4 6 5.1
mannose 0.8 5 3.8 4.0
3.4 3.6
Total pentoses 2.9 4 3.0
2.0 3 2.0 2.4
arabinose
pentose

xylose 0.4 2
rhamnose 0.4 1
fucose 0.1 0
0.0 2.0 4.0 6.0 8.0 10.0
Cake Honey biscuits Butter biscuits
Amount of NSP in % of DM
Figure 1. Amount of non-starch polysaccharides in Jerusalem artichoke powder (A);
total amount of non-starch polysaccharides in pastry products (B).

All pastry products used contain the mostly arabinose and xylose, representing pentose sugars,
which occur in flour as pentosans, or more precisley arabinoxylans. These polysugars influence
on quality parameter of flour, such as water absorption. With increasing JAP concentration
moisture of pastry products significantly increases, which can be explained by increasing
amount of arabinose follow by increased product’s water–holding capacity. Xylose is
decreasing in cakes (14 %), honey biscuits (30%) and butter bicuits (22 %) with added JAP in
concentrations 50 % and negatively affects the gas retention and loaf volume in the products.
However, the water holding capacity of the dough and the freshness of the pastry products
made of wheat and JAP are much dependent upon their content of NSP, mostly of arabinose
and xylose [1].

CONCLUSION
1. The addition of Jerusalem artichoke powder significantly (pd0.05) influenced the amount
of non-starch polysaccharides in pastry products.
2. Glucose, galactose, mannose, arabinose, xylose, rhamnose, fucose are found as the non-
starch polysaccharides monomers in Jerusalem artichoke powder and in pastry products
with Jerusalem artichoke powder.
3. Pastry products with added 30 % Jerusalem artichoke powder are permitted to call “source
of DF”.

REFERENCES
[1] Kays S. J. & Nottingham S. F. 2008. Biology and Chemistry of Jerusalem Artichoke Helianthus
tuberosus L. Boca Raton, CRC Taylor & Francis Group. p. 459.
[2] Boros D. & Aman P. 2009. Total Dietary Fiber. In: Shewry P. R. & Ward J. L. (Eds.). Healthgrain
Methods. Analysis of Bioactive Components in Small Grain Cereals. Inc. AACC International, St.
Paul, Minnesota, U. S. A, 167 – 176.
[3] Nielsen S. S. 2010. Food Analysis. Fourth edition. Springer, New York, p.602.

2150
 Characterization of cookies formulated with rice and black bean extruded flours

Bassinello, P. Z.a; Freitas, D. De G. C.b; Ascheri, J. L. R.b; Takeiti, C. Y.b; Carvalho, R. N.a; Koakuzu, S.
N.a; Carvalho, A. V.c

a
Embrapa Rice and Beans, Santo Antônio de Goiás, Brazil (pzbassin@cnpaf.embrapa.br;
rosangela@cnpaf.embrapa.br; selma@cnpaf.embrapa.br)
b
Embrapa Food Technology, Rio de Janeiro, Brazil (daniela@ctaa.embrapa.br;
ascheri@ctaa.embrapa.br; cristina@ctaa.embrapa.br)
c
Embrapa Eastern Amazon, Belém, Brazil (anavania@cpatu.embrapa.br)

INTRODUCTION
White rice and common beans are considered staple food in Brazilian diet as an important
source of good protein quality and energy. However, few actions are made with the byproducts
released from the rice milling or bean processing industries. Recently this concern has been
increasing in the country since other successful examples can be observed in different parts of
the world and our consumers are becoming more demanding for food diversity and quality.
Extrusion cooking technology is increasingly being used for the development of food
ingredients in order to promote the use of industrial wastes.
In order to utilize valuable components of the broken rice and the aged bean grains, which have
low commercial value, to produce different bakery products, this study aimed to investigate the
addition of a rice and black bean pre-gelatinized flour by extrusion on cookies formulations.

MATERIALS & METHODS

Mixtures of rice and peeled black bean (70:30) or whole black bean (60:40) were extruded on
Inbramac extruder in order to produce a pre-gelatinized flour (PBF and WBF respectively).
Four cookies samples (15% and 30% PBF; 15% and 30% WBF replacing corn starch) were
evaluated on nutritional composition, anti-nutrient contents, texture (TA-Hdi texture analyzer -
Stable Micro Systems, Surrey, UK) and color (Color Quest XE) measurements, and appearance
and global acceptability by 104 consumers on 9-point hedonic scales. The thiamin (vitamin B1)
and riboflavin (vitamin B2) were evaluated by HPLC according to the European Standard
methods. Tannins concentration was determined based on Deshpande and Cheryan (1985)
method description, using a methanol extraction and vanillin reaction with a catechin standard
curve. Phytate content was determined according to Haug and Lantzsch (1973) using a
spectrophotometer (Femto) calibrated to visible absorbance range. The rice-black bean pre-
gelatinized flour was also characterized for some of those parameters. The data were
statistically analyzed and significant differences among samples were assessed using ANOVA
(p<0.05).

RESULTS & DISCUSSION

The final products showed an interesting nutritional composition. The final formulations
presented higher carbohydrates contents especially when whole bean grain was used in the
flour. So, the cookies can be considered a good source of energy. The rice and bean cookies
had an average of 435 Kcal/100g of energetic value, which is around 9% lower than control,

11th International Congress on Engineering and Food - Athens, Greece, 2011 2151
probably due to their less fat content. This is considered a high value and so, the cookies can be
applied in the diet as energetic products. The protein content was significantly higher (about 2-
fold) for all samples in comparison to control. Based on Brazilian Ordinance nº 33, January 13,
1998, the average protein content found for rice and bean cookies (3.64%) allows us to affirm
that these products are a source of protein for children, because they attend the demanded
minimum percent of 20% of reference Recommended Daily Ingestion for each 100 g of food.
The lipid content was lower in the rice-bean cookies than control, especially for 30% of corn
starch substitution and when whole beans were used. Vitamin B content was slightly changed.
There was no tannin content detected in the final products and the level of phytate was very
low with no significant difference between products. This observation indicates that extrusion
process contributed to reduce anti-nutritional factors. There was significant difference between
products for some colour parameters. The sensory analyses (Table 1) showed that it can be
considered that consumers “liked lightly” the cookies with 15% and 30% PBF and 15% WBF.
Cookies formulated with WBF had lower acceptance by consumers in terms of appearance and
only those with 30% WBF had an intermediate score (”neither like, neither dislike”) for global
acceptability. Regarding texture, the cookies prepared with 30% WBF had their hardness and
brittleness increased. In general, when a higher amount of both flour (PBF and WBF) was
applied it was observed an increase of cookie hardness and brittleness values.

Table 1. Average liking scores (± standard deviations) for cookies

containing rice and black bean extruded flour.
Appearance Overall liking
15% PBF 6.14±1.84ª 6.25±1.92ª
30% PBF 6.21±2.01ª 6.17±1.70ª
15% WBF 4.33±2.17b 6.09±2.17a
30% WBF 4.41±2.22b 5.39±2.15b
PBF: rice and peeled black bean pre-gelatinized flour;
WBF: rice and whole black bean pre-gelatinized flour.

CONCLUSION
The obtained results indicate that both PBF and WBF flours may be adequate for cookies
formulating, although they need to be optimized, especially when whole bean grain was used in
the flour, regarding their effect on sensory characteristics. The cookies using rice and black
bean extruded flours can be considered a good source of protein, fibre, energy and presented
reduced anti-nutritional factors and lipid content, making possible to diversify the application
of by-products generated by rice and black bean processing industries.

REFERENCES
[1] Brasil, Ministério Da Saúde. Agência Nacional de Vigilância Sanitária. Portaria nº 27 de 13 de janeiro
de 1998. Disponível em: < http://e-legis.anvisa.gov.br/leisref/public/showAct.php?id=97>. Acesso
em: 18 outubro 2010.
[2] Deshpande, S.S.; Cheryan, M. (1985). Evaluation of vanillin assay for tannin analysis of dry beans.
Journal of Food Science, 50, 905-910.
[3] Haug W, Lantzsch H (1983) Sensitive Method for the Rapid Determination of Phytate in Cereals and
Cereals Products. Journal of Food Agricultural 34, 1423-1426.

2152
 Isolation of lactic acid bacteria in Marajoara cheese, Amazon, Brazil

Hamilton Mendes de Figueiredo, Cleidiane Gonçalves e Gonçalves, Paula Carolina de Moura Guimarães,
Adilson Mendes de Figueiredo Júnior

Engeneering Of Food, University Of Pará (hamiltonmendes1@hotmail.com)

INTRODUCTION
The Amazon, for its climatic and environmental conditions, has great agricultural potential,
with opportunities to excel in agribusiness, as supply center for domestic and foreign markets,
generating income and jobs. Here the regional cheeses are still produced with artisanal methods
in micro-enterprises typically family members, resulting in poor quality of products available
to consumers, and could affect their health. This problem stems primarily from inefficient
methods used in the cleaning of equipment and maintenance of the final product, besides the
lack of selected microorganisms and pure for use in the production of such cheeses.

Marajoara cheese, a typical product of Marajo-Amazon-Brazil, has been gaining market share
rapidly, which can be seen from this large consumption. So it becomes necessary to establish a
standard of identity and quality of this product, which may fix the minimum quality
requirements to be met for this cheese. The problem is that due to a craft like this is done,
hardly the quality standard will be in accordance with the law. This is mainly due to lack of
pasteurization of milk results in the presence of fecal bacteria that contaminate the product
during the steps of obtaining this. The development of these microorganisms in cheese leads to
a number of defects in the product, in addition to changes in flavor of it.

The objective this work is isolation and characterization of bacteria in Marajó cheese to former
a typical bank of lactic cultures in the Amazon.

MATERIALS & METHODS

The methodology was to collect three samples of cheese with one day and a serum when it is
still dripping cheese on a micro-industry in the region of the Marajó Archipelago followed by
transport to the microbiology laboratory of the Federal University of Para in Belem. The
samples were plated in deep agar medium M17 (Terzaghi and Sandino, 1975) and MRS and
incubated at 32 ° C for 72 hours. After this period was selected plates containing between 25
and 100 colonies for performing Gram staining and catalase test (Speck, 1984). The selected in
the previous tests were tested for hydrolysis of arginine (Silva, 2007), growth at 40 and 45 ° C
in skim milk and reconstituted (LEITE, 1993), gas production in MRS medium and growth at 4
and 6.5% salt in broth Bacto Lactose (Holt, et al 1994).

11th International Congress on Engineering and Food - Athens, Greece, 2011 2153
RESULTS & DISCUSSION
The results (Table 1) showed similar numbers of cells isolated on MRS or M17.

Table 1: Microbial count (log CFU / g or ml) and standard deviation

Amostra Ágar MRS Ágar M17
A <1,00 ± 0,000 <1,00 ± 0,000
B 6,38 ± 0,026 6,39 ± 0,013
C 4,42 ± 0,598 5,67 ± 0,039
SA 5,69 ± 0,013 4,95 ± 0,069
A, B and C: samples of cheese
SA: serum sample

Among the 20 isolates, 16 were able to hydrolyse arginine, 20 produced gas when MRS
medium, 19 grew at 40 °C, 10 grew at 45 ºC and 20 were able to grow at 4 and 6.5% salt. It
can be concluded that the 20 colonies analyzed, seven showed characteristics of Enterococcus,
eight of Lactococcus , two presented themselves as Streptococcus and three did not fit into any
of the characteristics of lactic acid bacteria.

CONCLUSION
Based on the results we can conclude it was possible to isolate 17 colonies with characteristics
of lactic acid bacteria. We also observed that most of lactic acid bacteria isolated were of the
genera Enterococcus and Lactococcus, which may mean that these are the genres that
contributed most significantly to the flavor of the cheese Marajó

REFERENCES
[1]Brasil. 1996. Regulamento técnico de identidade e qualidade de queijos. Ministério da
Agricultura do Abastecimento e da Reforma Agrária. Portaria nº 146 de 07 de março.
[2]Carvalho, J. D. G. 2007. Caracterização da microbiota lática isolada de queijo de Coalho
artesanal produzido no Ceará e de suas propriedades tecnológicas. Tese (Doutorado em
Tecnologia de Alimentos). Universidade Estadual de Campinas. Campinas. 154p.
[3]Finotelo, N. A. 1981. Melhoramento de tecnologia na produção e conservação do queijo
marajoara. Tese (Mestrado em Tecnologia de Alimentos). Universidade Estadual de Campinas.
Campinas. 113 p.
[4]Terzaghi, B. E.; Sandine, W. E. 1975. Improved medium for lactic streptococci and their
bacteriophages. Appl. Microbiol. 29, 807-813. 1975

Acknowledgments: FADESP – UFPA – PARÁ - BRASIL

2154
 The physico-chemical and microbiological aspects in ice-cream of buffalo milk added for

fiber food

Gerla C. B. CHINELATEa, Dorasílvia F. PONTESb, Roberto R. de A. BEZERRAc

a
UFCG, Pombal, Brazil (gerla@ccta.ufcg.edu.br)
b
UFC, Fortaleza, Brazil (dora@ufc.br)
c
UFCG, Pombal, Brazil (robson-aveiro@hotmail.com)

INTRODUCTION
Functional foods are now among the great progress made by man in order to promote and
provide health and quality of life. These foods, which naturally brings benefits to health have
been developed recently by taking advantage of recent knowledge acquired by engineers, food
technologists, chemists, nutritionists and health professionals. (CRAVEIRO e CRAVEIRO,
2003). [1]

Flaxseed is a plant food shop offering potential benefits for cardiovascular health by being an
important source of Į-linolenic acid (omega 3) and lignans, a class of phytoestrogens. Chitosan
- a naturally occurring biopolymer, found in the shells of crustaceans and other natural sources,
is composed of repeating units of D-glucosamine. It is considered a dietary fiber, since it has a
chemical structure very similar to cellulose, is also not digested by digestive enzymes
(MUZZARELI, 1996). [2]

Buffalo milk has higher concentrations of fat, protein, total solids and some minerals in relation
to bovine milk. For this reason, great importance is the transformation of food into their
products, since its peculiar composition offers high performance industry.

In this context, we developed this study in order to associate a functional ice cream formulation
based on buffalo milk supplemented with flaxseed and chitosan, in order to obtain products
with an alternative source of fiber, analyzing the physical and chemical interactions, chemical
and microbiological.

MATERIALS & METHODS

The first formulation was developed without the addition of flaxseed meal and chitosan, then
the others were added as an ingredient in Chitosan percentage set at 2%. This value was
defined by preliminary testing and balanced with the addition of flaxseed meal in different
proportions (0%, 5%, 10% and 15%), maintaining the standard 5% fat, 12% SNGL 15% sugar,
2% chitosan and 62% water, coded SQL-0 (ice cream with 2% chitosan and 0% flaxseed),
SQL-5 (ice cream with 2% chitosan and 5% flaxseed), SQL -10 (ice cream with 2% chitosan
and 10% flaxseed) and SQL-15 (ice cream with 2% chitosan and 15% flaxseed). For Physical,
physicochemical and chemical properties: moisture, protein, ash, lipids, carbohydrates, pH,
total acidity and iodine. For microbiological analysis was performed for Salmonella detection,
enumeration of coagulase-positive Staphylococcus and fecal coliform (BRASIL, 2001). [3]

11th International Congress on Engineering and Food - Athens, Greece, 2011 2155
RESULTS AND DISCUSSION
The ice creams were analyzed and processed results are presented in the table below.

Table1. Results of physico-chemical, physical and chemical properties.

Samples Moisture Proteins Ashes Carbohydrates Lipidis

SQL-0 70,17 ± 0,2a 6,49 ± 0,40a 0,97 ± 0,01a 13,57 ± 1,21a 4,80 ± 0,53a

SQL-5 63,00 ± 0,24b 6,55 ± 0,15a 1,10 ± 0,02a 13,82 ± 1,32a 5,24 ± 0,54a

SQL-10 60,29 ± 0,18c 6,69 ± 0,34a 1,21 ± 0,02a 14,43 ± 1,20a 5,40 ± 0,41a

SQL-15 60,19 ± 0,22d 7,00 ± 0,57a 1,30 ± 0,01a 12,78 ± 1,89a 6,40 ± 0,68a

The ice cream had moisture between 60.19 to 70.21%. This large variation is due to the
considerable addition of flaxseed meal to the formulations of ices (5%, 10% and 15%),
increasing the soluble solids content, resulting in a more consistent product. The protein
content ranged from 6.49 to 7.00%, increasing with the addition of flaxseed oil, thus improving
the nutritional characteristics of food. The ash content ranged from 0.97 to 1.30%, these
proportions guaranteed by adding the highest percentage of flaxseed. The increase of ash
content means a higher content of minerals in food, of great importance to their quality. The
percentage of carbohydrate was determined ranging from 12.78 to 14.43%, remaining in the
samples of four formulations developed a pattern of sugar content did not differ significantly at
5% of each other. The lipid content ranged from 4.80 to 6.40%. Due to the total replacement of
hydrogenated vegetable fat by oil meal in the formulation SQL-15, we observed a significant
increase in content of lipid fraction in the product. This fact justified by the presence of a
higher percentage of oil meal in the formulation

CONCLUSION
The application of flaxseed meal and ice cream in the processing of chitosan in different
proportions studied showed satisfactory results in relation to the physical-chemical,
microbiological, and nutritional and technological fit into the concepts of functional foods.

REFERENCES
[1] CRAVEIRO, A.A.; CRAVEIRO A.C.; QUEIROZ, D.C. Quitosana: A fibra do futuro. 1ª Edição,
281p. Fortaleza/CE, Editora Eletrônica Sandro Vasconcelos, 2003.
[2] MUZZARELI, R. A. A.; TANFANI, F.; EMANUELLI, M.; MARIOTTI, S. N-
(CABOXYMETHYLIDENE) Chitosans: Novel Chelating polyampholytes obtained from chitosan
glyoxylate. Carboydr. Res., v. 107, p. 199 – 210, 1983.
[3] BRASIL. Resolução RDC nº 12 de 02 de janeiro de 2011. Agencia Nacional de Vigilância Sanitária.
ANVISA, 2011.

2156
 Process Optimisation of Egg Replacer in Sponge Cake Baking

Levan Maia, Tom Nortonb, Weili Lia, Brijesh Tiwaria, Charles Brennana

a
Department of Food, Manchester Metropolitan University, Hollings Faculty, Manchester, UK
b
Department of Engineering, Harper-Adams University College, Shropshire, UK

INTRODUCTION
Heath risk associated with the consumption of eggs [1] and consumer preference for vegan diet
had lead researchers to investigate for egg replacers. The partial or total substitution of egg in
cake formulation appears interesting for people who suffer from cholesterol-diseases. The
replacement of egg in bakery products such as cake is a challenging task. Presence of egg
white proteins plays a significant role in foam formation by allowing the incorporation of large
volumes of air into the batter. Subsequent baking allows trapped air bubbles to expand leading
to an increase in the volume of cake after the coagulation of egg proteins during baking [2].
Response surface methodology has previously been used to optimise the processing variables
in partial or complete replacement of egg in cakes. For example, Arozarena et al., [3] employed
central composite design to investigate the effect of several ingredients on physicochemical
characteristics of lupine. The objective of this study was to optimise the baking process
parameters such as mixing time, baking time and temperature using Box-Benhken design.

MATERIALS & METHODS

Cakes were prepared using a standard recipe as outlined by AACC. Cakes samples were
analysed for texture, colour values (L*,a*,b*), specific volume, specific gravity, symmetry of
cake samples as per AACC. The effects of the three independent processing parameters on
quality parameters were investigated using Box-Behnken designs. The independent variables
were; X1 (4 – 10 min), X2 (160 – 200 oC) and X3 (10 – 30 min). Experimental data from the
Box-Behnken design was analysed and fitted to a second-order polynomial model.
3 3
[1]
0 ¦
Y E  E X  E X2 
i 1
i i ¦
i 1
iiE X X
i ¦¦
i j i 1
ij i j

where Y is the predicted response, ȕ0, ȕi, ȕii and ȕij are the intercept, linear, quadratic and cross
product coefficients. Xi and Xj are independent variables.

RESULTS & DISCUSSION

SG, texture, L*, a*, b* and Sym responses fluctuated between 0.74 to 0.64, 194.06 to 300.12,
50.03 to 77.30, 4.04 to 15.60, 16.86 to 33.64 and 0.15 to 1.80 respectively. The models
presented showed high regression coefficients (R2 > 0.80) for SG and colour values (L*, a*,b*)
whereas low regression coefficients (R2 = 0.47) laws observed for Sym of the cakes. The
predicted models were found to be significant with p values <0.0001 for SG, colour values (L*,
a*,b*) and 0.0039 for sym respectively.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2157
 a) b)
200 200
0.68 213 229
0.68 0.67
245
0.70
190 190
Baking temperature (oC)

180 180
0.71
221 213
0.68
170 170

0.67
0.69 221 237
0.66 205
160 160
10 12 14 16 18 20 10 12 14 16 18 20

o
Figure 1. Contour plots showing the effect of baking temperature ( C) and baking time (min) at mixing
time of 7 min on specific gravity (a), texture (b) of cakes.

Both linear and quadratic models were significant (p<0.05) for SG, texture, L* and b* whereas
both linear and cross product models were significant for a*. The statistical analysis showed
that the interaction among parameters was insignificant for SG, texture and sym (p<0.05). A
significant decrease in lightness value was observed with an increase in baking temperature
and time with similar increase in a* value. The second-order polynomial models obtained in
this study were utilized for each response in order to determine the optimum conditions. These
regression models are valid only in the selected experimental domain of mixing time, baking
temperature and time. This region was selected on the basis of product quality, as higher
baking temperature caused a sponge cake with a peaked centre and insufficient cooking time
led a collapse centre cake. The optimised condition shows that the optimum mixing time was 7
(range 6-8) min. The best baking condition for an optimum symmetry are in range between
170-183 oC with a baking time of 12.30-15.30 min respectively. However, control cakes
prepared from fresh egg were superior compared to the cakes prepared from egg replacer at
optimised baking conditions.

CONCLUSION
Response surface methodology using Box-Behnken design was found to be en effective
technique to investigate the impact of the mixing time, temperature and cooking time. The best
baking conditions for the cake with egg replacer was 7 min mixing time with baking
temperature of 180 oC and baking time of 14 min (12.3 – 15.30 min).

REFERENCES

[1] Shi, Z., Yuan, B., Zhang, C., Zhou, M., & Holmboe-Ottesen, G. (2011). Egg consumption and
the risk of diabetes in adults, Jiangsu, China. Nutrition, 27(2), 194-198.
[2] Çelik, I., Yilmaz, Y., IsIk, F., & Üstün, Ö. (2007). Effect of soapwort extract on physical and
sensory properties of sponge cakes and rheological properties of sponge cake batters. Food
Chemistry, 101(3), 907-911.
[3] Arozarena, I., Bertholo, H., Empis, J., Bunger, A., & de Sousa, I. (2001). Study of the total
replacement of egg by white lupine protein, emulsifiers and xanthan gum in yellow cakes.
European Food Research and Technology, 213(4-5), 312-316.

2158
 Obtaining functional fermented beverages by using the kefir grains.

Balabanova T.a, P. Panayotovb

a
Department “Technology of milk and dairy products”, University of food technology, Plovdiv, Bulgaria
(tbg_georgieva@yahoo.com)
b
Department “Technology of milk and dairy products”, University of food technology, Plovdiv, Bulgaria
( panayotov_p@yahoo.com)

INTRODUCTION
The increased interest to industrial dairy manufacture bring to full use of milk component, a
fuller and more rational use of separate secondary raw materials from processing. This
rationalization correlated from one side to ensure sustainability on production and high quality
of products, and from other side environmental protection through better utilization of
productive resources through the development and implementation of effective technological
systems to dairies for manufacturing of secondary raw materials [1]. It is known to obtain
fermented by kefir grains milk with high biological value. Obtaining large quantities of milk
serum from the cheese manufacturing and separately after membrane processing-filtrate is a
prerequisite for searching a different ways for their recovery.
The aim of this article is to obtain, examine and analyze fermented by kefir grains beverage
from whey, produced in the manufacture of Bulgarian brine cheese and ultrafiltrate.

MATERIALS & METHODS

The raw materials accompanying the experimental part are-milk, whey and ultrafiltrate. Milk is
used like a control sample, the whey is separated after the production of Bulgarian brine
cheese, cooled and stored refrigerated at t = 4-6qC. Filtrate obtained by using membrane
technology-ultrafiltration. The fermentation process for whey and filtrate are lead at 20-22qC
with 10% kefir grains and duration for 360 minutes. The experiments are realized in off-line
mode with ultrasound sensors of type UST40Ɍ/UST40R from Nippon Ceramic Company [2,3].

RESULTS & DISCUSSION

The dynamics of the change in pH during fermentation in raw materials (milk, whey and
filtrate), inoculated with kefir grains. Depending on the qualitative and quantitative
composition of raw materials and existing microflora are observe a gradual variation of pH in
the process of lactic fermentation in common cultivate of lactic acid bacteria and yeast (Fig.1).
Protein content of the raw materials also reflects on the changes in pH value. The control milk
sample is characterized by a higher buffering capacity, due to higher levels of protein
substance (3,4%). From the data in Figure 1 is observed gradual decrease in pH during
fermentation. Up to 120 minutes to reach pH 5,18, which coincides with izoyonic point of milk
and then are destabilize the casein. The filtrate and whey as opposed to the control sample
characterized by low buffering capacity (no protein substances), which allows for a short time
(20 minutes) the formation of small amounts of lactic acid, cause lowering the value on pH at
5,00 to 5,20.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2159
 Fig.1. Variation of pH during fermentation by kefir 0
grains 30
60
7 90
6 120
5 150
4 180
pH 210
3
240
2
270
1
300
0 330
filtrate whey milk
360

The duration of the fermentation process of milk is about 4 hours, but for whey and filtrate is
45 minutes. Characteristic of the control milk levels at the end of the fermentation process for
pH ranging from 4,5 to 4,6 and reached about 300-360 minutes, but for others raw materials
that period is five times shorter.
Non-contacts ultrasound method is beside on the effect of reflection. When passing through a
medium the ultrasound reduces its intensity, leading to its weakening. The absorption of
ultrasound energy in which ultrasound is spread depends on the characteristics of the medium-
density and elasticity. Thus, the ultrasonic signal carries information about the characteristics
of the medium between the transmitter and the receiver.

a b
Figure.6. Experimental data from measurements of beverages beginning coagulation-pH 5,2 (a), end of
fermentation-pH 4,5 (b)

CONCLUSION
The experimental technology and received laboratory results can be summarized several
important conclusions, one of which is the deployment of technologies for utilization of by-
products of milk production and seek alternative ways to create beverages milk-based foods
with certain nutrients and biological properties.

REFERENCES
[1] ɏɪɚɦɰɨɜ Ⱥ.Ƚ., ɋ. ȼ. ȼɚɫɢɥɢɫɢɧ, „ɋɩɪɚɜɨɱɧɢɤ ɬɟɯɧɨɥɨɝɚ ɦɨɥɨɱɧɨɝɨ ɩɪɨɢɡɜɨɞɫɬɜɚ”, Ɍɨɦ 5, 2004
[2] Shopov, N., R. Ilarionov, I. Simeonov, H. Kilifarev. Non-contact ultrasound method for identification
of yogurt according to its butter content. Proceedings of the International Conference on Computer
Systems and Technologies, CompSysTech’09, Rousse, Bulgaria, 18-19 June 2009, pp. I.1-1 – I.1-6.
[3] N. Shopov, R. Ilarionov, I. Simeonov. „Generating symptoms' spaces by wavelets for automatic
classification of yogurt according to its butter content”, Scientific works of University of Food
Technologies, Plovdiv, volume LVI, issue 2, 2009.

2160
 Effect of Synthesis Conditions of Short-Chain Fructooligosaccharides to Obtain High
Yield and Volumetric Productivity

Roberto Vegaa, María Elvira Zúniga-Hansena,b

a
School of Biochemical Engineering, Pontificia Universidad Católica de Valparaiso, Valparaiso, Chile
(rovegapa@gmail.com)
b
Regional Centre for the Study of Healthy Foods (CREAS), Valparaiso, Chile (mzuniga@ucv.cl)

INTRODUCTION
Short chain Fructooligosaccharides (sc-FOS) are a mixture of 1-kestose (GF2), nystose (GF3)
and 1F-fructofuranosylnystose (GF4), which are regarded as prebiotics since the middle of 90’s.
This recognition has allowed them to increase their demand in the food industry. Currently,
they are produced from sucrose by the action of fructosyltransferases, but with commercial
enzyme formulations of low cost have not been widely reported.
In this paper, we report the optimal conditions for production of sc-FOS from sucrose in order
to obtain high percentage of 1-kestose in syrups using a commercial enzyme preparation as a
source of food grade fructosyltransferase.

MATERIALS & METHODS

The commercial enzyme formulation, Rohapect®, was obtained from AB Enzymes GmbH
(Comercial Dimerco Ltda., Chile). Experiments were carried out in 20 mL of sucrose solution
in acetate buffer (50 mM, pH 5.5) and in stirred at 150 rpm. The conditions of temperature,
initial concentration of sucrose and enzyme concentration were studied. Sc-FOS and other
sugars were analyzed by HPLC.
The responses were yield (YP/S, g sc-FOS/100 g initial sucrose), 1-kestose in sc-FOS (ıGF2,
g/100 g sc-FOS) and volumetric productivity (QP, g/L.h), which were reported at 3 h reaction
time. The differences were statistically significant at p-values <0.05.

RESULTS & DISCUSSION

Experimental design results are shown in Table 1. Responses are presented at 3 h of reaction
time, when ıGF2 of all the experiences was higher than 70%. Higher yields were possible to
obtain, but in such cases were observed further transformation from GF2 to GF3. ıGF2 varied
with reaction conditions from 72 to 87.7%. Higher results of ıGF2 were achieved in trials 1, 8,
14 and 15 under different experimental conditions.
From a Pareto chart for each response for all the possible effects (Figure not showed) was
observed that the enzyme concentration (linear effect) had the greatest influence on the three
responses. The second effect on YP/S and ıGF2 was the temperature (quadratic effect), followed
by the initial concentration of sucrose (linear effect) and the interaction between temperature
and initial concentration of sucrose. The second effect on YP/S and ıGF2 was the temperature
(quadratic effect), followed by the initial concentration of sucrose (linear effect) and the
interaction between temperature and initial concentration of sucrose.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2161
Table 1. Full factorial central composite design of three variables with natural and coded units
Variables Coded variables Qp
YP/S
sc-FOS
sc-FOS ıGF2
T Sucrose Enzyme
Run x1 x2 x3
(°C) (M) (UT/mL)
1 50 2.1 4.2 -1 1 -1 42.9 ±0.4 87.7±0.1 102.9±1.0
2 50 1.7 6.6 -1 -1 1 59.9±1.1 72±0.0 115.8±2.2
3 60 1.7 4.2 1 -1 -1 44.2±0.8 83.5±0.1 85.4±1.6
4 55 1.9 7.4 0 0 1.682 62.2±0.5 67.6±0.0 134.8±1.1
5 55 2.2 5.4 0 1.682 0 47.3±0.1 83.5±0.8 121±0.3
6 55 1.9 5.4 0 0 0 51.7 76.4 112.1
7 60 1.7 6.6 1 -1 1 55.4±0.9 73.9±1.5 107.1±1.7
8 63.4 1.9 5.4 1.682 0 0 41.2± 1.6 84.8± 0.7 89.3± 3.6
9 55 1.9 5.4 0 0 0 52.5 76.9 113.7
10 60 2.1 6.6 1 1 1 57.7±0.2 72±0.3 138.5±0.5
11 60 2.1 4.2 1 1 -1 47.4±0.1 81.9±0.3 113.8±0.4
12 50 2.1 6.6 -1 1 1 54.8±1.3 79.9±0.1 131.6±3.0
13 55 1.6 5.4 0 -1.682 0 60.4±1.0 73.7±0.5 107.2±1.7
14 55 1.9 3.4 0 0 -1.682 45.8±0.0 84.9±0.1 99.3±0.1
15 46.6 1.9 5.4 -1.682 0 0 45.8±0.0 84.1±0.1 99,2±0.1
16 50 1.7 4.2 -1 -1 -1 51.4±0.1 82.1±0.3 99.3±0.3
17 55 1.9 5.4 0 0 0 53.1 77.6 115.1
(Mean ± standard deviation, n=2)

The second effect on productivity was the initial concentration of sucrose (linear effect),
followed by temperature (quadratic effect) and, finally, the interaction between temperature
and initial concentration of sucrose. The other effects were negligible at the 0.05 level of
significance and pooled into error. The highest p-value of significant effects was 0.014. The
ANOVA showed that the models are highly significant, which is evident from the low values
of probability of the null hypothesis. The lack of fit of models was not significant.

CONCLUSION
It was possible to determine the optimum condition for temperature, initial concentration of
sucrose and enzyme concentration in the reaction medium to obtain high percentage of 1-
kestose in sc-FOS from sucrose. The temperature was 49.6 °C, the initial concentration of
sucrose was 1.89 M and the enzyme concentration was 5.3 TU/mL. The sc-FOS YP/S reached
48% based on initial concentration of sucrose; ıGF2 reached 81% based on sc-FOS and QP
reached 103 g/L.h, all they under the above conditions at 3 h reaction time. The experimental
values to prediction values were 94.8% for yield of sc-FOS, 100.9% for 1-kestose in sc-FOS
and 95.1% for volumetric productivity.
The results indicate that the commercial enzyme formulation produces high content of 1-
ketoses, and presents a potential as biocatalyst in view of an industrial process for the
conversion of sucrose to syrup with high content of 1-ketose.

2162
Effect of pH culture on growth and fatty acid profile of Lactobacillus plantarum bacteria

C.Soto

Centro Regional de Estudios en Alimentos Saludables, Valparaíso, Chile

Escuela de Ingeniería Bioquímica. Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
(carmensoto@creas.cl)

INTRODUCTION
Bacteria such as Lactobacillus are used in the food industry to produce fermentable vegetable;
also, they are recognized as probiotics [3]. Some of these microorganisms are capable of
producing conjugated linoleic acid (CLA) [4], an isomer of linoleic acid, which is considered a
good fat and is used in prevention and control of cardiovascular disease and cancer treatment,
among others [2]. An important variable in microbial growth is the pH, affecting among other
things, the specific growth rate and formation of secondary products.
The aim of this study was to determine the effect of pH on growth and fatty acid profile of L.
plantarum, when this strain is grown in a supplemented medium with oils rich in linoleic acid,
in order to establish the viability of obtaining prebiotic biomass with a nutraceutical fatty acid.

MATERIALS & METHODS:

Lactobacillus plantarum NRRL - B4496 was donated by ARS-USDA. The culture medium
used was Man-Rogosa-Sharpe (MRS) including 2.67 mL/L of corn oil or grape seed oil.
Lactobacillus culture was carried out in an aerobic environment by batch. Three pH were
tested: 5.5, 6.5 and 7.5. A phosphate buffer (200 mM) was used to keep the pH. Microbial
growth was determined by spectrophotometry and by gravimetry. Glucose consumption was
determined using a specific enzymatic kit.
Intracellular and extracellular fatty acid profiles were determined after reaching a steady state.
Lipids were extracted using isopropyl alcohol and n-hexane from the culture media; then, lipids
were methylated in a derivatization process. Lipids from biomass were trans-methylated from
the cell. Methyl fatty acids were recovered with n-hexane and injected into the gas
chromatograph (Perkin Elmer Clarus 600) with FID detector.

RESULTS AND DISCUSSION

Figure 1 shows the effect of pH on Lactobacillus plantarum growth and on the glucose uptake
when the culture was supplemented with grape seed oil (Figure 1a) and corn oil (Figure 1b). As
it is possible to observe in both cases the maximum cell concentration was the same using
either a buffer pH 5.5 or 6.5 (3.2 g/L and 2.7 g/L using grape seed and corn oil respectively).
After 20 hours of fermentation, L.plantarum reached a stationary phase of growth at pH 6.5,
whereas at pH 5.5 this phase was achieve after 25 hours. For glucose decrease, a similar
behavior was observed with both pHs, with a total consumption after 20-23 hours. The specific
growth rate increase from 0.089 h-1 to 0.141 h-1 when the pH increased from 5.5 to 6.5. When
pH of 7.5 was applied growth was not observed.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2163
 4,0 25 4,0 25
3,5 3,5
3,0 20 20
Biomass(g/L)

3,0

glucose(g/L)
2,5 15 2,5 15
2,0 2,0
1,5 10 1,5 10
1,0 5 1,0 5
0,5 0,5
0,0 0 0,0 0
0 10 20 30 40 0 10 20 30 40
Time(h) Time(h)

Figure 1: Effect of pH culture on L.plantarum growth. Culture conditions: MRS medium supplemented
with grape seed oil (a) or corn oil (b). 2.67 mL / L; 37ºC; pH: 6.5 ({), 5.5 (š)

As it is possible to observe, in both cases (corn and grape seed oil supplementation)
Lactobacillus plantarum is able to adapt to different pHs.
Regarding the effect of pH on fatty acid profile, vaccenic and CLA fatty acids were observed
only inside the microorganism. If grape seed oil was incorporated, a similar CLA content was
observed when pH 5.5 or 6.5 were used, obtaining about 20% of this fatty acid. In the case of
LA a higher content is reported in depleted culture media. It is important to mentioned that LA
hydrogenation by microorganism enzymatic system have as final product stearic acid passing
through vaccenic acid generation; also, some ruminal microorganism are able to produce CLA
from oleic and vaccenic acids by '9-desaturase enzyme [1]. Due to this fact, these results
suggest a mechanism active by oil richest presence and satisfactory growth pH.
When comparing corn and grape seed oil, the last one is the best according L. plantarum
growth and intracellular CLA concentration.

CONCLUSION
pH during the culture of Lactobacillus plantarum bacteria can affect the growth rate and the
fatty acid profile. These effects depend markedly on the culture medium used.

REFERENCES
[1] Banni, S., Angioni, E., Murru, E., Carta, G., Melis, M.P., Bauman,D., Dong, Y. & Ip, C.2001.
Vaccenic Acid Feeding Increases Tissue Levels of Conjugated Linoleic Acid and Suppresses
Development of Premalignant Lesions in Rat Mammary Gland. Nutrition and Cancer, 41(1&2), 91-97
[2] Bhattacharya, A., Banu. J., Rahman. M., Causey. J. & Fernandes, G. 2006. Biological effects of
conjugated linoleic acids in health and disease. Journal of Nutritional Biochemistry. 17,789-810.
[3] Maragkoudakis, P.A., Zoumpopoulou, G., Miaris, C., Kalantzopoulo, G., Pot, B., & Tsakalido, E.
2006. Probiotic potential of Lactobacillus strains isolated from dairy products. International Dairy
Journal, 16,189–199.
[4] Ogawa, J.; Kishino, S., Ando, A., Sugimoto, S.; Mihara, K., & Shimuzu, S. 2005. Production of
conjugated fatty acids by lactic acid bacteria. Journal of Bioscience and Bioengineering, 100, 355-64.

2164
 Quality decay and viability of Lactobacillus acidophilus free and encapsulated in buffalo milk yogurt

A.S. Shoji a; A.C. Oliveirab; M.A. Trindadea; O. Freitasb, M. Thomazinia,

R.J.B. Heinemanna; C.S. Favaro-Trindadea,*
a
Universidade de São Paul.. Faculdade de Zootecnia e Engenharia de Alimento. Rua Duque de Caxias Norte, 225,
Pirassununga-SP, Brazil, CEP 13593-900 *Corresponding author: carmenft@usp.br
b
Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Avenida do Café, s/n°,
Ribeirão Preto-SP, Brazil, CEP 14040-903

INTRODUCTION

The viability of the microbial content and the general quality of many probiotic-containing products is still under
question. Several reports have shown that the survival and viability of probiotic bacteria is often low in yoghurt [1],
resulting in counts lower than 107–108 CFU/g of the daily. Innovative technologies have investigated the instability
problems of these microorganisms. Microencapsulation is one of the approaches that have presented excellent results
[2, 3]. Thus, the aim of this work was to evaluate the quality decay and the viability of L. acidophilus in buffalo milk
yogurt.

MATERIAL & METHODS

Materials: A citrus pectin GENU R (REF. 13596) esterification degree of 68% (CP Kelco, Brazil) and Bovine casein
(Katuffmann, Germany) were used as encapsulating agents. The Lactobacillus acidophilus Lac-04 culture was kindly
donated by Danisco (Brazil) in a pure, freeze-dried form and kept at -18oC. Buffalo milk and a traditional culture for
yoghurt (L. bulgaricus and S. thermophilus from Christian-Hansen, Brazil) were used to prepare the yoghurts.
Microencapsulation: The microcapsules were prepared in accordance with the method described by Oliveira et al.
[13] and freeze dried.
Application of microcapsules in yoghurt: For yoghurt preparation, buffalo milk was pasteurised as described by
Marcatti et al. [4], cooled to 40°C and inoculated with a traditional culture of yoghurt and (1.5g/100g of yogurt) for
probiotics (1.5g/100g of yogurt). The filling was carried out in packs of 100 mL incubated at 40°C until the product
reached pH levels of 5.0 and 4.5. Four pilot-scale yoghurt manufacturing protocols (designated T1, T2, T3, and T4)
were carried out (n=4). Yoghurts T1 were manufactured with a traditional culture of yoghurt followed by adding
encapsulated L. acidophilus and fermenting it to pH 5.0. Yoghurts T2 were manufactured with a traditional culture of
yoghurt by adding free L. acidophilus fermenting it to pH 5.0. Yoghurts T3 were manufactured with a traditional
culture of yoghurt by adding encapsulated L. acidophilus and fermenting it to pH 4.5. Yoghurts T4 were manufactured
with a traditional culture of yoghurt by adding free L. acidophilus and fermenting it to pH 4.5. All yoghurts were kept
in refrigeration for 28 days for storage evaluation. The samples were analysed by pH, acidity and viability of probiotic
cultures during 28 days for refrigerated storage.

RESULTS & DISCUSSION

Table 1 presents physicochemical data for buffalo yoghurts prepared with the addition of L. acidophilus. There was an
overall decline in pH during refrigerated storage of all yoghurts, being more pronounced in the early days. The
yoghurts prepared with free form of the probiotics (T2 and T4) resulted in a more accentuated decline of pH than the
yoghurts prepared with microencapsulated probiotics. Considering the final fermentation pH or initial pH in the storage
of the yoghurts with microencapsulated probiotics of 5.0, the decrease was from 5.02 to 4.17 within 28 days of
refrigerated storage and 4.9 to 4.0 in the yoghurts with free cultures at the same conditions. The same profile was
observed for yoghurts with a pH at 4.5 at the beginning of the storage conditions. Thus, it can be inferred that the
microencapsulation of L. acidophilus in buffalo yoghurt results in lower post-acidification values, independent of the
pH at the beginning of storage. Also, the pH, both 4.5 and 5, did not show a significant effect on the variables

11th International Congress on Engineering and Food - Athens, Greece, 2011 2165
 evaluated. The results from the pH decline in the free and encapsulated forms can be an indication that the
encapsulation process results in lower metabolic activity.

Table 1. pH and titrable acidity of yoghurts containing probiotics during refrigerated storage for 28 days.
Day 1 Day 14 Day 28
Treatment ** pH Acidity pH Acidity pH Acidity
T1 5.02±0.01aA 0.82±0.015aA 4.14±0.01 aB 0.94±0.015aA 4.17±0.01 aB 0.96±0.022aA
T2 4.99±0.01aA 1.23±0.013bA 3.99±0.01 bB 1.35±0.011bA 4.01±0.01bB 1.40±0.005bA
T3 4.53±0.01bA 0.93±0.020aA 4.14±0.01 aB 0.98±0.020aA 4.20±0.02 aC 1.06±0.018aA
T4 4.51±0.01 bA 1.25±0.031bA 3.97±0.02 bB 1.36±0.018bAB 4.01±0.02 bB 1.45±0.018bB
* Results presented as a mean (n=3) standard error of mean (n=3)
a,b,c: Means with different small letters in the same column differ (p < 0.05) A,B,C: Means with different capital
letters in the same line differ (p < 0.05)
** T1: Yoghurt with encapsulated L. acidophilus fermenting to pH 5.0. T2: Yoghurts with free L. acidophilus
fermenting to pH 5.0. T3: Yoghurts with encapsulated L. acidophilus fermenting to pH 4.5. T4: Yoghurts with L.
acidophilus free fermenting to pH 4.5

The viability of the probiotic cultures in the buffalo yoghurt was also determined during the storage period of 28 days
as shown in Table 2. For L. acidophilus in free and microencapsulated forms, there was a decay in the number of
viable cells throughout the refrigerated storage period, reaching values between 4.4 and 4.9 log CFU/g for the
probiotics in free form (T2 and T4) and 7.9 and 7.5 log CFU/g for the encapsulated probiotics (T1 and T3) after 28
days. Treatment T1, with a pH of 5.0 at the beginning of storage, presented the highest number of viable cells after
storage time, while T2 presented the lowest counting among the treatments. These results differ from those from the
evaluation of the survival of L. acidophilus at pH 1 and 3. This suggests that the microencapsulated L. acidophilus
resisted the processes employed (encapsulation, fermentation and cooling) was not affected by intrinsic conditions (pH,
acidity and the potential for oxidation-reduction) of yoghurt, as well as those of storage (refrigeration temperature) and
had a better performance than the free probiotic culture.

Table 2. Effect of final fermentation pH and form of probiotics on the viability of L. acidophilus (counts of L.
acidophilus in log cfu g-1) in buffalo yoghurt
Treatment** Storage (days) *
1 14 28
T1 9.24±0.02aA 9.14±0.04aA 7.99±0.01aB
T2 9.34± 0.02aA 8.54±0.02bB 4.38±0.07dC
aA bB
T3 9.20±0.01 8.46±0.07 7.51±0.06bC
aA aA
T4 9.31±0.03 9.04±0.04 4.95±0.04cB
*
Results presented as a mean (n=3) r standard error of mean (n=3)
a,b,c
: Means with different small letters in the same column differ (p < 0.05) A,B,C: Means with different capital letters in
the same line differ (p<0.05)
**
T1: Yoghurt with encapsulated L. acidophilus fermenting to pH 5.0. T2: Yoghurts with free L. acidophilus
fermenting to pH 5.0. T3: Yoghurts with encapsulated L. acidophilus fermenting to pH 4.5. T4: Yoghurts with L.
acidophilus free fermenting to pH 4.5

These results suggest that microencapsulation by complex coacervation followed by lyophilisation provides sufficient
protection for the L. acidophilus culture in yoghurt made from buffalo milk.

CONCLUSION

Yoghurts prepared with microencapsulated L. acidophilus result in lower values of post-acidification and higher
culture viability during cold storage in a number sufficient to promote therapeutic effects on consumer health superior
to 107 CFU/g over the 28 days. Thus, yoghurt prepared by buffalo milk is a good vehicle for the incorporation of
probiotics.

2166
 Supercritical fluid extraction with modifier of antioxidant compounds from jabuticaba

(Myrciaria cauliflora) by-product: economic viability

Rodrigo N. Cavalcanti, Priscilla C. Veggi, M. Angela A. Meireles*

A

A LASEFI/DEA/FEA (School of Food Engineering)/UNICAMP (University of Campinas) – R. Monteiro

A Lobato, 80; 13083-862, Campinas, SP, Brazil (meireles@fea.unicamp.br)
B
INTRODUCTION
Jabuticaba (Myrciaria cauliflora) is a Brazilian grape-like fruit with extensive occurrence in
the country. A part of production of these fruits is explored by local populations, but the
majority is wasted during harvest and processes. Thus, the application of innovative
technologies such as supercritical fluid extraction (SFE) processing by-products is important to
obtain high quality products adding value to these products. Besides, by-products processes
currently represents an increasing niche of market mainly due to its ecological, economic and
social implications. Indeed it is necessary a critical analysis of chemical composition and
economic viability of the extracts obtaining in order to evaluate the industrial applicability [1].
The objective of this work is to evaluate the feasibility of antioxidants recovery by supercritical
fluid extraction (SFE) with co-solvent using different conditions of pressure and temperature.

MATERIALS & METHODS

Jabuticaba jelly residue was provided by Santa Maria farm (Campinas, São Paulo, Brazil)
grinding in knife mill (Tecnal TE-631 model series 01071, Piracicaba, São Paulo, Brazil),
packed in plastic bag, and stored in domestic freezer (Metalfrio, HC-4, Sao Paulo, Brazil) at -
18 °C. The SFE assays were carried out using a system with a 415 cm3 extraction vessel (3.4 ×
10-2 m of diameter and 37.5 × 10-2 m of height) performed in two temperatures (323 and 333
K) and three pressures (10, 20, and 30 MPa) using ethanol as modifier at 20% v/v. The global
yield (X0) was calculated as ratio of extract mass (mextract) and mass of raw material (mRM)
loaded in the extraction vessel as shown in the equation 1:
m extract
%X 0 ˜ 100 (1)
m RM
Radical scavenging activity using DPPH (2,2-diphenyl-1-picrylhydrazyl) was performed
according to the method of Kordali et al. [2] at 517 nm in UV-vis spectrophotometer (Hitachi,
model U-3010, Tokyo, Japan). Antioxidant activity of the extracts was calculated by
scavenging ability (%SA) at 0.2 mg/cm3 extract concentration according to equation 2 as
follows:

% SA

Abs Control  Abs Sample u 100 (2)
Abs Control
SuperPro Designer 6.0® was used for process simulation and economical evaluation. The main
costs that compose the manufacturing costs (COM) are similar to the ones described by Turton
et al. [3].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2167
RESULTS & DISCUSSION
The increase in temperature shown to have a significantly influence increasing the global yield
of the supercritical extracts but decreasing antioxidant compounds recovery. Manufacturing
costs of the antioxidant compounds (COMCA) showed high variability with the variation of
temperature, pressure and size of the cell extraction. The COMCA increased with temperature
probably due to energetic costs. The increase in pressure showed a decrease (10 to 20 MPa)
followed by a pronounced increase in manufacturing costs (20 to 30 MPa). This phenomenon
is closely related to antioxidant compounds recovery and energetic costs.

Table 1. Global yield, antioxidant activity and cost of manufacturing of extract and antioxidant
compounds obtained by SFE with ethanol at different operational conditions.

Global Antioxidant COMX0 (US$/kg) COMCA (US$/kg)

Sample Yield Activity
(% d.b.) (% d.b.) 0.05 m3 0.3 m3 0.05 m3 0.3 m3

323K-10MPa 8,321 13,01 18,17 8,99 139,70 69,12

323K-20MPa 15,20 15,10 18,13 8,98 120,08 59,48
323K-30MPa 10,15 7,971 18,18 9,01 228,08 113,04
333K-10MPa 25,46 8,754 18,37 9,26 209,85 105,78
333K-20MPa 20,85 12,35 18,42 9,28 149,20 75,17
333K-30MPa 24,72 4,188 18,40 9,28 439,38 221,60

CONCLUSION
Thus, evaluating extract and the antioxidant compounds yield as well as the manufacturing
costs for each condition of temperature and pressure it is possible to conclude that the
extraction condition at 323 K and 20 MPa had the highest yield of antioxidant compounds and
lower cost of manufacturing (COM-CA) being selected as the best choice by supercritical fluid
extraction with ethanol to obtain antioxidants from jabuticaba jelly residue.

REFERENCES
[1] Arts I.C.W. & Hollman P.C.H. 2005. Polyphenols and disease risk in epidemiologic studies.
American Journal of Clinical Nutrition, 81(1), 317–325.
[2] Kordali S., Kotan R., Mavi A., Cakir A., Ala A., & Yildirim A. 2005. Determination of the chemical
composition and antioxidant activity of the essential oil of Artemisia dracunculus and of the
antifungal and antibacterial activities of Turkish Artemisia absinthium, A. dracunculus, Artemisia
santonicum, and Artemísia spicigera essential oils. Journal of Agricultural and Food Chemistry, 53,
9452–9458.
[3] Turton R., Bailie R. C., Whiting W. B. & Shaeiwitz J. A. 1998. Analysis, synthesis, and design of
chemical process. 1st ed., Prentice Hall, Upper Saddle River, NJ, 1998. p. 848.

2168
 Microencapsulation of sacha inchi (Plukenetia volubilis L.) oil with zein

Sócrates Quispe-Condoria,b, Marleny D.A. Saldañab

a
School of Food Engineering, Universidad Peruana Unión, Ñaña, Lima, Perú (socrates@upeu.edu.pe)
b
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta
(marleny@ualberta.ca)

INTRODUCTION
The consumption of polyunsaturated fatty acids (PUFA’s) has gained great importance due to
the variety of health benefits. Fish, flax and algae are the commonly sources of PUFA’s.
Recently, sacha inchi seeds (Plukenetia volubilis L.) has been commercialized as an alternative
source. Sacha inchi tree is a millenarian legacy, of the Inca civilization, that is being
extensively cultivated in the Peruvian Amazon. Sacha inchi seeds are valued for their high oil
(35-60%) and protein (27-33%) content. Furthermore, its oil is an excellent source of
polyunsaturated fatty acids, mainly linolenic acid (C18:3 Ȧ-3) and linoleic acid (C18:2 Ȧ-6)
[Hamaker et al., 1992]. However, one of the major drawbacks of oils containing a high amount
of PUFA’s is their rapid oxidation. The use of encapsulation technologies to retard the
oxidation of these oils has drawn considerable attention [Sanguansri & Augustin, 2007]. Zein,
the prolamin fraction of corn protein, has long being recognized for its coating ability and
mechanical and barrier properties. Encapsulation of sacha inchi oil using zein as a carrier has
not been reported in the literature. Therefore, the objective of this study was to evaluate the
physical properties of sacha inchi microcapsules produced by spray drying. The microcapsules
were analyzed for their particle yield, microencapsulation efficiency, flowing properties,
particle size distribution and morphological characteristics.

MATERIALS & METHODS

Factorial design (Table 1) was used to investigate the physical properties with respect to zein
(x1) and sacha inchi oil (x2) concentrations. The microencapsulation of sacha inchi oil and the
physical chemistry analysis were carried out according to Quispe-Condori et al. [2011].

Table 1. Experimental matrix (actual values and coded levels) for the factorial design and particle yield,
microencapsulation efficiency (MEE) and flowing properties of sacha inchi oil - zein microcapsules
Particle Flowing properties
Zein Oil Zein:oil Mean diameter
# Yield MEE (%) Bulk density Hausner
(%) (%) Ratio Carr Index (Pm)
(%) (kg/m3) ratio
1 10 (0) 1 (1) 10:1 76.93 79.95±0.59 206.91±2.93 1.48±0.02 32.37±0.97 n.d.
2 10 (0) 0.5 (-1) 20:1 66.57 84.50±0.84 203.37±10.51 1.52±0.08 34.10±3.55 n.d.
3 8 (-1) 1 (1) 8:1 62.55 78.81±0.09 245.26±5.20 1.41±0.05 29.17±2.26 207.44
4 12 (1) 1 (1) 12:1 75.59 88.53±0.38 243.66±20.65 1.55±0.10 35.16±4.22 22.63±1.49
5 8 (-1) 0.5 (-1) 16:1 59.15 84.98±0.14 274.78±11.60 1.48±0.06 32.28±2.87 84.46±16.01
6 12 (1) 0.5 (-1) 24:1 92.73 86.23±0.16 209.21±10.38 1.61±0.03 37.93±1.25 52.85±13.77
7 10 (0) 0.75 (0) 13.3:1 80.56 90.41±0.74 194.73±19.35 1.55±0.19 34.69±8.72 n.d.
8 10 (0) 0.75 (0) 13.3:1 78.41 86.60±0.65 216.35±17.09 1.58±0.11 36.42±4.30 40.64±28.45
9 21 (-) 0.75 (0) 28:1 92.84 86.54±0.85 183.70±9.82 1.51±0.12 33.45±5.54 n.d.
n.d. = not determined

11th International Congress on Engineering and Food - Athens, Greece, 2011 2169
RESULTS & DISCUSSION
Particle yield, microencapsulation efficiency, flowing properties and particle size distribution
of sacha inchi oil microcapsules are presented in Table 1. The response surface contour plots
for particle yield and microencapsulation efficiency are presented in Figure 1.
1 1 94
Sacha inchi oil concentration (%)

Sacha inchi oil concentration (%)

90
80 90
70 86
60 82
50 78

0 0

-1 -1

-1 0 1 -1 0 1

Zein concentration (%) Zein concentration (%)

Figure 1. Estimated contour plot in the factorial design experiment obtained by plotting the sacha inchi
oil and zein concentration for (a) Particle yield, (b) Microencapsulation efficiency.

Morphological analysis showed that there were not significant differences in the morphologies
of the sacha inchi oil microcapsules at different zein:oil ratios (Figure 2)

Run 4 Run 6

Figure 2. Scanning electron micrographs of microcapsules at different operational conditions.

CONCLUSION
It was demonstrated that particle yield was significantly affected by the zein concentration,
while microencapsulation efficiency and flowing properties were not affected by zein and
sacha inchi oil concentration. Particle size distribution and morphology of the microcapsules
depend on the zein:flax oil ratio.

REFERENCES
[1] Hamaker B.R., Valles C., Gilman R., Hardmeier R.M., Clark D., Garcia H.H., Gonzales A.E.,
Kohlstad I., Castro M., Valdivia R., Rodriguez T. & Lescano M. 1992. Amino acid and fatty acid
profiles of the Inca Peanut (Plukenetia volubilis L.). Cereal Chemistry, 6(4), 461-463.
[2] Sanguansri L. & Augustin M.A. 2007. Microencapsulation and delivery of Omega-3 fatty acids. In:
Shi, J. (Ed.), Functional Food Ingredients and Nutraceuticals: Processing Technologies. Taylor &
Francis, Florida, USA.
[3] Quispe-Condori S., Saldaña M.D.A., Temelli F. 2011. Microencapsulation of flax oil with zein using
spray and freeze drying. LWT – Food Science and Technology, In press.

2170
 Encapsulation of Curcumin Loaded Oil Droplets by Cryotropic Gel Formation from

O/W Emulsion

Kyuya Nakagawa1*, Nataporn Sowasod2, Tawatchai Charinpanitkul3, Apinan Soottitantawat3

and Wiwut Tanthapanichakoon3
1
Research Centre for Nano-Micro Science and Engineering, University of Hyogo, Japan
2
Nanoscience and Technology Program, Graduate School, Chulalongkorn University, Thailand
3
Center of Excellence in Particle Technology, Faculty of Engineering, Chulalongkorn University,
Thailand (*e-mail to Nakagawa: nakagawa@eng.u-hyogo.ac.jp)

INTRODUCTION
Nowadays, nano-micro encapsulation is a key technology to stabilize active food ingredients such as
vitamin, polyphenol, peptide etc., and to put a controlled release function onto a matrix. Hydrogel is a
widely accepted encapsulant, as naturally used in many traditional gel foods. These gel structures usually
possess nano-scale holes formed by polymeric chains, and the sizes of these holes are determined by the
characteristics of the polymers and the degree of gel formation. It is a reasonable strategy to design a
controlled release system in a hydrogel by controlling the gelation manners. But it is not always easy to
realise it because of the difficulty of sol-gel transition management in an industrial process. The authors
believe that these problems would be overcome by using cryotropic gel formation where the network
formation could be controlled by a freezing operation. Cryotropic gelation is a sol-gel transition induced
by the concentration increase of the substrates due to the dehydration by freezing (ice formation).
Lozinsky et al. have reported a lot of intensive studies on cryotropic gelation [1, 2]. This cryotropic
gelation would also be a useful tool for oil encapsulation, however, the number of reports on the relevant
topics is still lacking. The present study was aimed to encapsulate an oil phase into cryogel matrices
obtained from a ternary system of chitosan, ț-carrageenan, and carboxy methylcellulose sodium salt
(NaCMC). Triolein oil that contained curcumin was selected as the oil phase, and o/w emulsions were
prepared with suspended chitosan (emulsifier). A suspension of ț-carrageenan and NaCMC was added to
this emulsion to convert them into cryogels after freezing. The obtained frozen cryogels were
subsequently freeze-dried to obtain dried gels that contain curcumin oil. Release behavior of curcumin
was investigated in an aqueous buffer solution, and the influence of freezing conditions on the release
characteristics was reported.

MATERIALS AND METHODS

A triolein solution containing 0.3 wt% of curcumin (oil phase) was mixed with a 3 wt% chitosan aqueous
suspension containing 2 wt% of acetic acid and 5 wt% of Tween80. The ratio of the oil phase to the
aqueous phase was set at 10 %(v/v), and this mixture was emulsified with a high-speed homogenizer at
12,000 rpm for 5 min. 5 %(w/v) of sodium chloride was added to the prepared o/w emulsion, then mixed
with another polymer suspension consisting of 1 wt% of polymer (ț-carrageenan and NaCMC mixture)
and 5% of sodium chloride. Two sets of the suspensions were prepared by varying the ratio of
carrageenan to NaCMC (set A: 1:9, set B: 4:6). The mixture of the prepared o/w emulsion and this
polymer suspension was homogenized at 12,000 rpm for 5 min to obtain a homogeneous colloidal
suspension. The prepared suspension was frozen by cooling with a contact plate heat exchanger at -40°C,
where the cooling rates were controlled, namely; -0.5, -1.0, -2.0 K/min, respectively. The obtained frozen
samples were subsequently freeze-dried for 24 hours. A piece of the cylindrical freeze-dried cryogel

11th International Congress on Engineering and Food - Athens, Greece, 2011 2171
sample (dia. 10 mm, height 10 mm) was set in a test tube with 5 ml of the buffer solution, and the test
tube was installed in a shaking bath where the temperature was maintained at 37ºC. The released amounts
of curcumins were evaluated by HPLC analysis to obtain release curves.

RESULTS AND DISCUSSION

First of all, we should note that the present formulations successfully enabled the preparation of
irreversible gel formation merely due to freezing. The freeze-thawed specimens were fragile, however,
they possessed enough strength to be removed from the test tube and held directly by hand. The obtained
freeze-dried cryogels were spongy and their structural hardness depended on their formulation and
freezing condition. These differences were appeared on their release characteristics as listed in Table 1.
The total released amount of curcumin (for four days) was almost determined by the freezing conditions.
They increased as increasing the cooling rates during freezing.

Table 1. Sample IDs and the total released amount of curcumin

Sample S-1 S-2 M-1 M-2 F-1 F-2
Mixed polymer suspension set A B A B A B
Cooling rate during freezing
-0.5 -0.5 -1.0 -1.0 -2.0 -2.0
[°C/min]
Total amount released
41.1 43.3 48.7 48.3 58.05 59.9
for 4 days [%]

The release curves, as depicted in Fig. 1, were mostly burst type curves. A curve obtained from F-2,
however, was first order release type. It was worth noting that S-2, M-2 and F-2 were prepared from the
same suspension, but their characteristics were greatly dependent on their freezing history. It means that
the gel network structures in the present cryogel system were greatly controlled by their gel formation
kinetics during sol-gel transition. And this structural modification consequently affected to the
diffusivities of the core ingredient (i.e. curcumin). This cryogel based technique holds big potential for
producing various types of hydrogel structures from a certain suitable formulation simply by changing
their processing parameters.
70

60

50
% Release

40

30 S-1
S-2
20 M-1
M-2
10 F-1
F-2
0
0 500 1000 1500 2000 2500 3000
Time [min]

Figure 1. Release curves of curcumin from freeze-dried cryogels (in PBS pH 7.4)

REFERENCES
[1] Lozinsky, V. I. et al. (1986) Acta Polymerica, 37, 142-146
[2] Lozinsky, V. I., & Damshkaln, L. G. (2000) Journal of Applied Polymer Science, 77, 2017-2023.

2172
 Effect of different ratios of maltodextrin/gelatin and ultrasound in the

microencapsulation efficiency of turmeric oleoresin

Cassia Roberta Malacridaa, Vânia Regina Nicoletti Telisa

a
São Paulo State University – UNESP, São José do Rio Preto, Brazil (cmalacrida@terra.com.br,
vanianic@ibilce.unesp.br)

INTRODUCTION
Encapsulation is a process in which one or more ingredients or additives are coated with a
small and edible capsule. This technique seems to be useful to solve problems regarding
limitations in the use of food ingredients, as turmeric oleoresin, which despite having
numerous advantages over the turmeric powder, is sensitive to light, heat, oxygen and pH
variations. The main emphasis of microencapsulation has been concentrated on improving the
encapsulation efficiency and extending shelf-life of the products. The properties of the wall and
core material as well as the emulsion characteristics and drying parameters are the factors that
can affect the efficiency of encapsulation [1].
The objective of this work was to study the influence of different ratios of maltodextrin/gelatin
used as wall materials and the effect of ultrasound application during the emulsification step on
the encapsulation efficiency of turmeric oleoresin by freeze drying.

MATERIALS & METHODS

Different concentrations of maltodextrin 10 DE (12.0 - 29.7 %) and bovine gelatin, bloom
value 240, (0.5 - 6.0 %) were dispersed in distilled water. The turmeric oleoresin was added to
the mixtures at a fixed ratio of 15 %, based on the mass of carrier materials, and emulsified in a
shear homogenizer for 15 min at 18,000 rpm. Sonication was applied by immersing samples in
an ultrasonic bath for 15 minutes. The homogenized mixtures were frozen at -38 ºC for 24
hours and freeze dried at < -40 °C for 48 hours. The encapsulation efficiency was calculated
based on the curcumin content [2] retained in the microcapsules after freeze drying. Moisture
[3] and solubility in water [4] of the encapsulated powder were also determined. All
experiments were carried out in duplicate and average values are reported.

RESULTS & DISCUSSION

Moisture contents varied from 1.1 to 4.1 % (wet basis) and it is possible to observe that the
samples with the highest gelatin content (6 %) presented the highest moisture percentages.
Regarding to solubility experiments, only the samples with higher gelatin percentage (6 %)
were not soluble in water within 5 minutes of agitation. The application of ultrasound was only
significant for the encapsulating matrix consisting of 18 % maltodextrin and 6 % gelatin,
reducing the time of solubilisation from 15.5 minutes (without ultrasound) to 12 minutes (with
ultrasound).
The increasing gelatin concentration in the polymeric matrix increased the encapsulation
efficiency of turmeric oil (p < 0.05), whereas the maltodextrin content in the wall material did
not show significant effect at a 5 % level of significance. The effectiveness of ultrasound to

11th International Congress on Engineering and Food - Athens, Greece, 2011 2173
improve encapsulation efficiency was more pronounced for a higher content of maltodextrin in
the wall material: for an encapsulant matrix formulated with 18 % maltodextrin and 6 %
gelatin, the encapsulation efficiency increased from (81.1 ± 0.4) % without ultrasound to (90.4
± 1.4) % when applying ultrasound. In the case of an encapsulant matrix formulated with 12 %
maltodextrin and 6 % gelatin, the increase in the encapsulation efficiency was lower: (77.6 ±
0.6) % without ultrasound to (80.6 ± 0.1) % when applying ultrasound.

Table 1. Influence of the wall material formulation on turmeric oleoresin encapsulation efficiency
Formulation Maltodextrin Gelatin Moisture* Solubilization time* EE%*
(%) (%) (wt %) (min)
1 20.6 0.9 1.8 ± 0.13 1.2 ± 0.20 57.8 ± 0.30
2 20.6 2.6 2.2 ± 0.12 3.0 ± 0.00 79.7 ± 0.30
3 28.1 0.9 1.2 ± 0.10 2.0 ± 0.13 50.8 ± 2.00
4 28.1 2.6 1.1 ± 0.07 3.6 ± 0.34 64.1 ± 0.30
5 19 1.8 1.5 ± 0.05 2.5 ± 0.09 58.6 ± 1.50
6 29.7 1.8 1.3 ± 0.06 3.8 ± 0.50 62.5 ± 2.50
7 24.4 0.5 1.4 ± 0.06 2.3 ± 0.11 65.3 ± 1.20
8 24.4 3 1.8 ± 0.00 1.3 ± 0.26 56.3 ± 3.10
9 24.4 1.8 1.4 ± 0.02 3.0 ± 0.00 54.8 ± 1.00
10 12 6 3.6 ± 0.04 15.1 ± 0.19 77.6 ± 0.40
11 18 6 4.1 ± 0.01 15.5 ± 0.22 81.1 ± 0.30
12** 12 6 4,0 ± 0.07 14.8 ± 0.15 80.6 ± 0.10
13** 18 6 4.2 ± 0.04 12.0 ± 0.40 90.4 ± 1.00
*Mean values ± standard error (n=2)
**Samples with ultrasound application

CONCLUSION
The encapsulation efficiency was significantly affected by gelatin content, indicating the
effectiveness of gelatin as an encapsulant for turmeric oleoresin. The application of ultrasound
in the emulsification step of turmeric oleoresin and maltodextrin/gelatin appeared to improve
emulsion quality and stability, increasing the curcumin retention in the freeze-dried powders.

REFERENCES
[1] Jafari S.M., Assadpoor E., He Y. & Bhandari B. 2008. Encapsulation efficiency of food flavours and
oils during spray drying. Drying Technology, 26, 816-835.
[2] Chauhan S., Singh B. & Agarwal S. 1999. Estimation of curcuminoids in Curcuma longa by HPLC
and spectophotometric methods. Indian Journal of Pharmaceutical Sciences, 61, 58-60.
[3] AOAC. 1995. Official methods of analysis of the Association of Official Analytical Chemists.
AOAC, Arlington.
[4] Wang Y., Lu Z., Lv F. & Xiaomei B. 2009. Study on microencapsulation of curcumin pigments by
spray drying. European Food Research Technology, 229, 391-396.

2174
 Encapsulation of Melissa Officinalis leaf’s active compounds in ȕ-cyclodextrin

and modified starch

Ioannis Mourtzinos1, Spyridon E. Papadakis2, Panagiotis Igoumenidis3 & Vaios T. Karathanos3

1
Apivita SA, Natural Products & Cosmetics, Koletti 3, 144 52, Metamorfosi, Athens, Greece
2
Laboratory of Food Packaging, Department of Food Technology, Technological Educational Institute of
Athens 12210, Egaleo, Athens, Greece
3
Laboratory of Chemistry & Physical Chemistry of Foods, Department of Nutrition & Dietetics, Harokopio
University, 17671, Kallithea, Athens, Greece

INTRODUCTION
Melissa officinalis (lemon balm) is a perennial herb in the mint family Lamiaceae, native to
southern Europe and the Mediterranean region [1]. Its main active compounds are phenolic acids,
which are well known antioxidants [2]. Phenolic acids also possess antibacterial, antiviral, and anti-
fungal properties, and are known to stimulate the immune and blood circulatory systems [3]. In
order to use lemon balm leaf extract as a nutraceutical, effective methods of extraction and
encapsulation of phenolic acids are required. The aim of this work was to prepare encapsulated
forms of lemon balm leaf extract in ȕ-cyclodextrin and modified starch in solid state. Another
objective was to verify the encapsulation via Differential Scanning Calorimetry (DSC) and to study
the stability of the free and encapsulated extract under inert and oxidative conditions.

MATERIALS & METHODS

A Semi-automatic extraction equipment for the simulation of a percolation of the type Timatic was
used. The encapsulated forms of lemon balm constituents in ȕ-Cyclodextrin (ȕ-CD) and in
modified starch were prepared. A Differential Scanning Calorimeter was employed to study the
thermo-oxidation stability of the samples. The antioxidant activity of the extract and also those,
which were extracted in presence of ȕ-cyclodextrin, was also studied with antioxidant radical
scavenging methodology.

RESULTS & DISCUSSION

The formation of inclusion complex of an extract in ȕ-CD can be confirmed by obtaining DSC
thermograms in an inert atmosphere for: (a) pure extract (b) pure ȕ-CD, (c) physical mixture of
extract with ȕ-CD, and (d) the encapsulated extract.
In Figure 1 the DSC oxidation curves of (a) pure lemon balm extract, (b) the inclusion complex of
lemon balm with ȕ-CD and (c) lemon balm encapsulated in modified starch are presented. The
exothermic peak initiated at 146°C for pure lemon balm is related to lemon balm’s constituent’s
oxidation. This peak was not present in the DSC scan of the lemon balm/ȕ-CD complex indicating

11th International Congress on Engineering and Food - Athens, Greece, 2011 2175
that constituents of lemon balm are protected from oxidation being inside the ȕ-CD cavity. Similar
results were obtained by DSC of the lemon balm/ MS sample under oxidative conditions at the
same temperatures, (Figure 1c), indicating that lemon balm’s constituents are also more stable
when it is encapsulated in MS.

Figure 1. DSC thermograms of (a) pure

lemon balm extract, (b) complex of
lemon balm/ȕ-CD and (c) lemon
balm/modified starch under oxidative
conditions.

The assessment of the in vitro antiradical activity showed that the antioxidant activity of the extract
increased when the extraction was performed in presence of ȕ-CD. The increment can be attributed
to the increment of the solubility of lemon balm constituents in water due to the inclusion complex
formation with ȕ-CD.

CONCLUSION
The encapsulated molecules are protected from oxidation, as depicted from oxidative DSC studies.
Therefore, the encapsulated forms can be used as additives to foods. Moreover the dissolution of ȕ-
CD in the extraction solvent increased the antioxidant activity of the obtained extract.

REFERENCES
[1] Kim S., Yun E.J., Bak J.S., Lee H., Lee S.J., Kim C.T., Lee J.H., Kim K.H. 2010. Response surface
optimised extraction and chromatographic purification of rosmarinic acid from Melissa officinalis leaves.
Food Chemistry 12, 521–526.
[2] Herodez S.S., Hadolinb M., Skergeta M., Kneza Z. 2003. Solvent extraction study of antioxidants from
Balm (Melissa officinalis L.) leaves. Food Chemistry 80, 275–282
[3] Dastmalchia K., Damien Dormana H.J., Darwisd Y., Laakso I., Hiltunena R. 2008. Chemical
composition and in vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract. LWT-
Food Science & Technology 41, 391–400.

2176
 Deployment of Response Surface Methodology to Optimize Recovery of Grape
(Vitis vinifera) Stem and Seed Polyphenols

Evangelia Karvela1, Dimitris P. Makris2, Nick Kalogeropoulos1, Vaios T. Karathanos1

1
Department of Science of Dietetics-Nutrition, Harokopio University, 70, El. Venizelou, 17671, Kallithea, Athens, GREECE
2.
Department of Food Science & Nutrition, University of the Aegean, 2, Mitr. Ioakim, 81400, Myrina, Lemnos, GREECE

INTRODUCTION

Industrial wine production is accompanied by the generation of large quantities of waste streams, including inorganic
material (e.g. bentonite clay) and by-products composed of bio-organic substances (skins, seeds and stems). In Europe,
it is estimated that 14.5 million tonnes of grape by-products are produced, on an annual basis, deriving from the
winemaking industry. Thus the increasing demand for environmentally compatible production, coupled with rising
operational and waste treatment cost, has started to move wine industry towards adoption of integrated waste
preventive approaches. The investigations carried out on the efficient retrieval of polyphenols from winery wastes have
mainly been focused on red pomace, which is characterised by relatively high burden in phenolics and pigments.
Grape stems, which represent a fraction of the total grape waste generated during the vinification process, is a tissue
that has been given relatively little attention, in spite of recent reports on its polyphenolic composition that appears to
incorporate substances not encountered in other by-products, e.g. flavonols and stilbenes, in addition to monomeric and
oligomeric flavanols. Furthermore, the polyphenolic content of stems was shown to be approximately 5.8% on a dry
weight basis, and therefore stems may be a source of bioactive phenolics that should not be overlooked.
Ethanol is a bio-solvent, which is food compatible, reusable and cheap and it has become the solvent of preference for
quite a few recent studies pertaining to the recovery of phenolics from various plant tissues. In the case of winery
wastes, there is a scarcity of data concerning the use of ethanol-based solvents for extracting phenolic phytochemicals.
This being the conceptual basis, the study presented herein provided some novel aspects pertaining to polyphenol
retrieval from grape stems, using factorial design and response surface methodology.

MATERIALS & METHODS

Stems and seeds from three widely cultivated wine grape varieties were chosen; one white (Savatiano), one red used
for white wine production (Moschofilero) and one red used for red wine production (Agiorgitiko). Extraction
procedure was carried out as described by [5]. A 23 full-factorial experimental design was used to identify the
relationship existing between the response functions and process variables as well as to determine those conditions that
optimised the extraction process in relationship with ethanol concentration, extraction time and pH. Several indices of
the polyphenolic composition, such as total polyphenol (TP) [4], total flavanol (TFl) [4], total flavone (TFn) [2],
proanthocyanidin (PC) [4] and their antioxidant capacity, such as Antiradical Activity (AAR) [1] and Reducing Power
(PR) [4], were determined as they are described to the related articles.

RESULTS & DISCUSSION

The experimental screening performed was designed to assess the influence of three factors (ethanol concentration, pH
and extraction time). The experimental data obtained showed a good fit with the equations, which most of them were
statistically significant (p=0.05). The trends revealed in each case were recorded in the form of three-dimensional plots
(Figs. 1-2), where on the left is illustrated the effect of simultaneous variation of pH and EtOH, and on the right the
effect of simultaneous variation of time and EtOH. The trends recorded in each case, as well as the discrepancies
revealed regarding the optimum EtOH level, suggested that optimisation of polyphenol recovery from tissues with
different polyphenolic composition should be based on case experimentation, and that there is not a universal model
describing the optimal conditions that should be deployed. From the optimisation process it became evident that the

11th International Congress on Engineering and Food - Athens, Greece, 2011 2177
 extraction of flavanols requires an EtOH level of 60%, as opposed to flavones, which can be efficiently recovered with
40% EtOH. For proanthocyanidins (PC), which represent flavanol oligomers and polymers, intermediate EtOH
concentrations ranging from 44.2 to 53.1% were the most satisfactory. Significant differences were also caused by
altering the pH. While flavanol extractions gave higher yields at pH 2-3, flavones were better extracted at pH 4.5-6.
The results for PC were absolutely consistent, indicating a pH 2 as the optimal. Such a consistency for PC was also
observed for the duration of the extraction, where for all samples the time required for optimal yields was 5 hours. On
the other hand, optimal extraction times for TFl and TFn were 1-3.5 and 1-5 hours, respectively, indicating that, in
general, extraction of higher TFn amounts required extended extraction time, compared with TFl.

A A Fig. 1: Response-surface plot showing the effect of EtOH /

pH (left) and EtOH / time (right) co-variance on the total
polyphenol (TP) yield. A, Moschofilero; B, Savatiano; C,
B B
Agiorgitiko (stems).

Fig. 2: Response-surface plot showing the effect of EtOH

C C / pH (left) and EtOH / time (right) co-variance on the
total polyphenol (TP) yield. A, Moschofilero; B,
Savatiano; C, Agiorgitiko (seeds).

Extraction of TP from Agiorgitiko seeds required higher EtOH levels compared with the Moschofilero and Savatiano
counterparts, suggesting that this tissue contained less polar substances. Further, it has been supported that increasing
pH values might enhance polyphenol solubility by promoting dissociation of the most acidic phenolic –OH groups,
which would render polyphenols higher solubility in a hydroalcoholic medium. Such a hypothesis would explain why
TP and TFl optimal extraction from Agiorgitiko seeds required higher pH. The time necessary to attain optimum levels
was, on average, shorter for PC (1.14 h) and almost the double for TFl (2.33 h). The correlations between antioxidant
activity and polyphenols differed for grape skins and seeds. Careful interpretation of the data given for stems shows
that in Moschofilero and Savatiano extracts, the only statistically significant correlations were those established
between PC concentration and PR, while for Agiorgitiko a significant link was found between PC and AAR. For
Moschofilero, the correlation of TP was also significant but low. The correlations established for grape seeds suggested
that in Moschofilero extracts, which contained higher amounts of TFl and PCs, links were statistically significant with
both AAR and PR, but for Agiorgitiko this held true only for the correlations of TP, TFl and PCs with AAR; for Savatiano
only TP and TFl gave high correlations with AAR. This outcome does not display any consistency and might indicate
that interactions among the various flavanol forms rather define the overall antioxidant potency.

CONCLUSION
The examination presented herein demonstrated that the set of conditions employed to optimise extraction of phenolics
from plant material may vary substantially, even for the same tissue originating from different varieties [3]. Grape
stems were shown to contain mainly flavanols and flavonol glycosides, but it appeared that flavanol oligomers and or
polymers (proanthocyanidins) define the antioxidant magnitude of the extracts obtained. The finding that extraction of
different polyphenol classes from grape stems requires different set of conditions might be of value in selective
recovery, for the generation of extracts enriched in particular components. Grape seeds were shown to contain mainly
flavanols and flavanol dimers, which rather define the antioxidant magnitude of the extracts generated. If seeds
originate from red pomace used in red wine production, then partial exhaustion of phenolics is to be anticipated. This
variation in the composition as a result of processing might fundamentally affect the polyphenolic profile and
consequently the conditions required for effective retrieval. These crucial differences in the conditions should be
carefully considered when extractions are not directed and recovery of as many phenolics as possible is sought.

2178
 Production of 1-octen-3-ol by Neurospora species isolated from beiju
in different culture medium

D. S. de Carvalhoa, A. P. Dionísioa, R. dos Santosa, S. Boguzs Jrb, H. T. Godoyb , G. M. Pastorea

a
Laboratory of Bioflavours, Department of Food Science, Faculty of Food Engineering (UNICAMP), P.O. Box 6121,
13083-862 Campinas-SP, Brazil (danisc31@gmail.com.)
b
Laboratory of Food Analysis, Department of Food Science, Faculty of Food Engineering (UNICAMP), P.O. Box 6121,
13083-862 Campinas-SP, Brazil

INTRODUCTION

Flavors and fragrances are key impact substances in the food and fragrance industry and these compounds are commonly
produced chemically. However, the growing market share of flavoured and fragranced products requires novel strategies for
aroma chemicals. Biotechnological options comprise single-step biotransformations, bioconversions and de novo synthesis
with microorganisms, plant cells and enzymes. Some fungal species are able to produce the unsatured alcohol 1-octen-3-ol,
known as “mushroom-like flavor” and “raw mushroom”, as a product of its secondary metabolism, in a procedure known as
de novo synthesis. One of these microorganisms is Neurospora sp., and some works showed a production of 1-octen-3-ol
using this specie (Pastore et al, 1995 and Yamauchi et al, 1991). In this context, the aim of this study was to evaluate the
production of 1-octen-3-ol in different culture medium by some Neurospora species.

MATERIALS & METHODS

Production of mushroom aroma

Four culture medium were used for the production of aroma: malt extract broth (50 g.L-1); yeast malt broth (YM:10 g.L-1 of
glucose, 5 g.L-1 of peptone, 3 g.L-1 of yeast extract and 3 g.L-1 of malt extract); fructose (50 g.L-1)/yeast extract (5 g.L-1);
and Czapeck medium standard, with some modifications (40.6 g.L-1 of NH4H2PO4,10 g.L-1 of K2HPO4, 5 g.L-1 of
MgSO4.7H2O, 5 g.L-1 of KCl, 0.01 g.L-1 of FeSO4.7H2O and 30 g.L-1 of sucrose).
Preparation of pre-inoculum and fermentation
Neurospora sp. strains were inoculated into slant tube of potato dextrose agar (PDA) at 30°C for 72h. After, 10 mL of sterile
water were added into tube and transferred for 250 mL Erlenmeyer flasks containing 50 mL of YM medium at 30°C, 200
rpm for 24h. Then the culture broth was filtered through a membrane of acetate filter (pore size: 0.45 μm) and the mycelia
were washed with sterile water. The inoculum occurred using 1 g of the biomass added in a flask containing 50 mL of each
medium described above and homogenized under sterile conditions using an Ultra-Turrax ® T18 (Ika, Wilmington, NC,
USA). The flasks were incubated on a rotary shaker at 30°C for until 144 h under agitation (200rpm). Each 24h samples
were collected and analyzed in gas chromatography (GC-FID).
GC-FID conditions and GC-MS conditions
The volatile compound, extracted with diethyl ether, was analyzed using an Agilent GC (Model 7890A) equipped with a
flame ionization detector (FID) and a HP-5 columm (Agilent Tecnhologies i.d. = 0.320 mm, length = 30 m, film thickness =
0.25μm, USA). Injector ( mode splitless) and detector were 250°C. GC-MS analyses were carried out in a GC-MS system
(Shimadzu GC-17A/QP-5000 high performance quadrupole, Japan) in the same conditions of GC-FID. Transfer line
temperature: 240°C, energy of impact: +70 eV, 35-350m/z). The identification of components was made by mass spectrum
an retention index agreed whit standards and comparing spectra (Adams, 2007 and NIST, 2005).
Statistical analysis

11th International Congress on Engineering and Food - Athens, Greece, 2011 2179
The data obtained were analyzed using ANOVA/Tukey (p<0.05). The statistical package used was StatisticaTM 8.0 data
analysis software by Statsoft, Inc.,USA.

RESULTS & DISCUSSION

As shown table 1 the analytical method was adequate for the analysis of 1-octen-3-ol.

Table 1. The concentration range, regression equations, R2, recovery, LOD and RSD for 1-octen-3-ol
Matriz Conc. range Regression equation R2 Recovery RDS range LOD LOQ
-1
(mg.L ) (%) % (mg.L-1) (mg.L-1)
Malt
1-40 Y=0.0163x - 0.0022 0.998 38.53 0.40 1.55
extract 3.52-8.74
YM 1-40 Y=0.0131x-0.0059 0.998 83.85 1.84-10.92 0.59 2.17
Fructose/
yeast 1-40 Y=0.0208x+0.0007 0.998 99.78 0.89-15,74 0.52 1.49
extract
Czapeck 1-40 Y=0.0366x+0.0295 0.994 43.42 2.68-10.51 0.57 2.02

Table 2. The table 2 shows the maximum yield of 1-octen-3-ol in each culture broth.
Culture broth Strain 1-octen-3-ol (ppm) Time of fermentation (h)
Malt extract LB12DSC 17.17a 48
Fructose/
LB13DSC 16.51a 96
yeast extract
YM LB13DSC 12.53 b 48
Czapeck LB13DSC 0.85 c 24
* Different letters represent statistically significant differences (p<0,05).

As shown in table 2, the malt extract broth was the most adequate medium for production of 1-octen-3-ol. Strain LB23DSC
produced 17.17 mg.L-1 of 1-octen-3-ol in malt extract broth followed the LB13DSC in fructose/yeast extract medium.

CONCLUSION

All Neurospora strains studied produced 1-octen-3-ol in the different mediuns tested. However, the high yields of this
compound occurred using malt extract broth.

REFERENCES

[1] Adams, R. P. (2007) Identification of Essential Oil Components by Gas Chromatography/Quadrupole Mass Spectrometry,
Allured Publishing Corp.: Carol Stream, Illinois, USA.
[2] Assaf, S., Hadar, Y., Dosoretz C.G. (1997). 1-Octen-3-ol and 13-hydroperoxylinoleate are products of distinct pathways in the
oxidative breakdown of linoleic acid by Pleurotus pulmonarius. Enzyme and Microbial Technology, 21, 484-490.
[3] Bicas, J.L., Silva, J.C., Dionísio, A.P., Pastore, G.M. (2010). Biotechnological production of bioflavors and functional sugars.
Ciência e Tecnologia de Alimentos, 30(1). 7-18.
[4] Rossi, S.C., Vandenberghe, L.P.S., Pereira, B.M.P., Gago, F.D., Rizzolo, J.A., Pandey, A., Soccol, C.R., Medeiros, A.B.P.
(2009). Improving fruity aroma production by fungi in SSF using citric pulp. Food Research International, 42 (4), 484-486.

2180
 Characterisation of a non-alcoholic beverage made of residues from king palm

(Archontophoenix alexandrae) industry

Karina Cardoso Tramonte, João Gustavo Provesi, Iolanda Moreira Dutra Albuquerque E Silva, Aureanna
Nairne Negrão Murakami, Marcelo Maraschin, Renata Dias De Mello Castanho Amboni And Edna
Regina Amante*

Department of Food Science and Technology, Federal University of Santa Catarina, Rodovia Admar
Gonzaga 1.346, 88034-001 Florianópolis, SC, Brazil
Corresponding author: Fax: (55) (48) 3721 99 43. Telephone: (55) (48) 3721 53 71. E-mail:
eamante@cca.ufsc.br

INTRODUCTION
The exploration of palm trees for the production of canned palmito generates large amounts of
solid residue because it requires the felling of the whole plant, where only the inner sheath is
exploited, and the leaves, the stems, and the external and the median sheaths are discarded [1].
The objectives of this study were: to propose a suitable procedure for obtaining and preserving
the juice of king palm leaf sheaths; to study the chemical composition of the juice; to develop a
non-alcoholic beverage with this juice and assess its acceptability by potential consumers.

MATERIALS & METHODS

The leaf sheaths were washed in potable water and immersed in 0.3 M citric acid solution to
prevent browning and then they were blanched by immersion in boiling water for 3 minutes for
enzyme inactivation. This material was pressed using a roller mill with stainless steel clamps.
For each 100 mL of juice, were added 5 g of citric acid and 12.5 mg of ascorbic acid.
Centesimal composition was determined according to the methods recommended by the
AOAC [2]. Total phenolic content (TPC) was determined by the Folin-Ciocalteu method [3].
The free radical scavenging capacity of the extracts of the leaf sheathes of king palm was
analyzed by the DPPH• (2,2-diphenyl-1-picrylhydrazyl radical) [4]. Identification and
quantification of phenolic compounds was performed by HPLC [5].

RESULTS & DISCUSSION

The extract, or juice, has high contents of carbohydrates and minerals. The total polyphenol
content was 19.91 ± 0.07 g GAE 100 g -1. The juice of king palm leaf sheaths showed EC50 of
3.50 ± 0.01 mg . g-1 DPPH. The DPPH radical scavenging capacity, represented by the
percentage of inhibition, was 86.54 ± 0.09 %. The interest in antioxidants from natural sources
has grown because of their high capacity to scavenge free radicals, linked to reduced risk of
several diseases and of their greater food safety in comparison with synthetic antioxidants,
butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Table 1 shows phenolic
compounds in juice from leaf sheath of king palm.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2181
Table 1. Phenolic compounds in juice from leaf sheath of king palm
Retention time Concentration
Phenolic compounds
(min) (ȝg/100 g - dry basis) a
Gallic acid 4.85 3219.87 ± 377.12
3,4-Dihydroxybenzoic acid 7.2 4170.90 ± 585.44
Chlorogenic acid 9.39 3393.58 ± 315.76
Syringic acid 11.30 1834.08 ± 151.16
Caffeic acid 12.90 10781.21 ± 916.87
a
Data are mean ± SD (n = 3).

CONCLUSION
The juice from king palm leaf sheathes showed interesting nutritional characteristics, with high
values of minerals, such as magnesium and potassium, and of total polyphenols and also high
DPPH free radical scavenging capacity, which suggest a high antioxidant activity. The
presence of caffeic, chlorogenic, 3,4-dihydroxybenzoic, gallic and syringic acids in this new
juice, suggest new studies about it functional properties.

REFERENCES
[1] Vieira, M. A., Podestá, R., Tramonte, K. C., Amboni, R. D. M. C., Simas, K. N., Avancini, S. R. P. &
Amante, E. R. (2009). Chemical composition of flours made of residues from the king palm
(Archontophoenix alexandrae) industry. Brazilian Archives of Biology and Technology, 52, 973-
980.
[2] AOAC. Association of Official Analytical Chemists (2005). Official methods of analysis, 18th ed.
Maryland, USA: The Association.
[3] Singleton, V. L. & Rossi, R. J. A. (1965). Colorimetry of total phenolics with phosphomolybdic-
phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16, 144-158.
[4] Brand-Williams, W., Cuvelier, M. E. & Berset, C. (1995). Use of a free radical method to evaluate
antioxidant activity. LWT – Food Science and Technology, 28, 25-30.
[5] Schuldt, E. Z., Bet, A. C., Hort, M. A., Ianssen, C., Maraschin, M., Ckless, K. & Ribeiro-do-Valle, R.
M. (2005). An ethyl acetate fraction obtained from a southern brazilian red wine relaxes rat
mesenteric arterial bed through hyperpolarization and NO-cGMP pathway. Vascular Pharmacology,
43, 62-68.

2182
 Composition of aroma compounds in fermented apple juice: effect of apple variety,

fermentation temperature and inoculated yeast concentration

Riekstina-Dolge R. a, Kruma Z. a, Karklina D. a, Seglina D. b

a
Latvia University of Agriculture, Faculty of Food Technology,
Liela iela 2, LV – 3001 (rita.riekstina@llu.LV)
b
Latvia State Institute of Fruit Growing, Graudu iela 1, Dobele, LV-3701 (dalija.seglina@lvai.lv)

INTRODUCTION
Apple juice is the raw material of different fermented drinks, like apple wine and cider. Aroma
plays significant role in the quality of cider, and composition of volatiles depends on used
technology, maturation and storage conditions [1]. Cider maturation significantly influenced
the chemical composition of cider distillates. The concentrations of ethyl esters of the major
organic acids of cider (lactic, acetic and succinic), as well as the contents of aromas produced
by the bacteria activity (2-butanol, 2-propen-1-ol, 4-ethylguaiacol and eugenol) increased with
increasing levels of cider maturation [2]. The aim of current research was to evaluate aroma
composition of fermented apple juice, depending on used apple variety, fermentation
temperature and yeast concentration in must.

MATERIALS & METHODS

For analysis two apple varieties ‘Auksis’ (Streif index 0,16, starch index 8,4) and ‘Lietuvas
Pepins’ (Streif index 0,24, starch index 6,2 ) were used. Apples were grown in the Latvia State
Institute of Fruit Growing and harvested in autumn 2010. Juice was obtained by press Voran
Basket press 60K. For stabilization of juice Tannisol (Enartis, Italy) were used. Fermentation
was performed using commercial Saccharomyces bayanis yeast EC-1118 (Lalvin, Canada)
(three concentrations in juice and two fermentation temperatures). Volatiles from apple juice
and fermented drinks were extracted using solid phase microextraction (SPME) using
divinylbenzene/carboxen/polydimethylsiloxane fiber (Supelco Inc., Bellefonte, PA, USA).
SPME parameters were: incubation time 30 min, extraction temperature 22±2 °C, extraction
duration 30 min, desorption 15 min, 250 °C. For the analysis of the SPME extracts, a Perkin
Elmer Clarus 500 GC/MS and a Elite-Wax ETR (60 m x 0.25 mm i.d.; DF 0.25 ȝm) was used.

RESULTS & DISCUSSION

A total of seven volatile compounds (total peak AU 2999,93 ×105) were detected in ‘Auksis’
apple juice and aroma is mainly composed of aldehydes (60,5%) and esters (35%). The main
volatile compounds of ‘Auksis’ is 2-hexenal (50,1 %), 1 butanol, 2- methyl acetate (13,1%)
and hexanal (10,5%). In ‘Lietuvas pepins’ juice, compared to ‘Auksis’ apple juice, higher
content of aroma compounds were detected (6 volatile compounds and total peak AU
4490,35×105). The main volatile compounds of ’Lietuvas Pepins’ apples juice were acetic acid
butylester (35,3%), acetic acid hexylester (24,2%) and 1 butanol, 2- methyl acetate (15,6%).
Some compounds found in apple juice were not possible to identify in fermented apple juices,
namely aldehydes hexanal and 2-hexenal and alcohols 2-hexen-1-ol. A total of 16 volatile

11th International Congress on Engineering and Food - Athens, Greece, 2011 2183
compounds were detected in ‘Auksis‘ apples fermented juice at the early stage of fermentation
and 13 at the 28 day of fermentation. As major groups of juice have been identified alcohols
(67, 5%), esters (31%) and acids (1,6%) after 8 day fermentation and alcohols (54,1%), esters
(42,8%) and acids (3,2%) after 28 day fermentation. Acetic acid, phenylethyl alcohol and 3-
methyl-1-butanol were characteristic compounds of yeasts. Among higher alcohols determined,
all the strains produced 3- methyl-1-butanol that is the major higher alcohol in wine [3,4]. n-
decanoic acid, hexanoic acid ethylester, decanoic acid ethylester, 9-Decenoic acid, ethyl ester
were detected after 8 day fermentation of ‘Auksis’, but were not detected after 28 day
fermentation. Whereas opposite tendency were observed for hexanoic acid hexylester and 8-
heptadecanol. They were detected after 28 day fermentation, but wasn’t at the early stage of
fermentation. Characteristic apple aroma compound acetic acid butylester were detected in
fermented juice only at the initial stage of fermentation. Hydroxyethylhydrazine was found as a
main volatile aroma compounds in all fermented apple juices. It is possible to identify higher
concentrations of apple juice aroma compounds in fermented juices with lower concentration
of inoculated yeast. Acetic acid, 1-butanol, 3-methyl phenylethyl alcohol found in fermented
juices, are typical yeast aromas aroma compounds. Others aroma develops during fermentation
process. The highest content of aroma compounds was detected in LP4 (with lower
concentration of yeast), followed by LP3 (fermented with lower concentration of yeast).
Content of acetic acid increased during fermentation, and the highest content was detected with
the lowest concentration of inoculated yeast.

CONCLUSION
The main aroma compound of ‘Auksis’ apple juice is 2-hexenal, whereas for ‘Lietuvas Pepins’
acetic acid butyl ester. Typical used yeast aroma compounds are acetic acid, phenylethyl
alcohol and 3-methyl-1-butanol. The main aroma compounds in fermented juices were 2-
hydroxyethylhydrazine, 3-methyl-1-butanol, and hexanoic acid ethyl ester, acetic acid hexyl
ester. Typical apple aroma compounds acetic acid hexylester and 1- hexanol better maintained
in juices fermented with lower concentration of yeast at lower temperature. Ethyl acetate and
acetic acid butylester concentration in fermented juice was higher in samples fermented at
higher temperature. Others esters group (octanoic and decanoic acid ethyl esters), that
developed during fermentation process, showed the highest concentrations in samples
fermented at lower temperatures.

REFERENCES
[1] Mangas J.J., Gonzalez M.P., Rodriguz, R., Blaco, D.(1996). Solidphase extraction and determination
of trace aroma and flavour components in cider by GC-MS. Chromotorapia, 42,101-105.
[2] Rodríguez Madrera R., Picinelli Lobo A., Mangas Alonso J. J. (2009). Effect of cider maturation on
the chemical and sensory characteristics of fresh cider spirits. Food Research International, 70-78.
[3] Romano, P., Capece, A., Serafino, V., Romaniello, R., Poeta, C., 2008. Biodiversity of wild strains of
Saccharomyces cerevisiae as tool to complement and optimizƝ wine quality. World J. Microbiol.
Biotechnol. 24, 1797–1802.
[4] Garde-Cerda´ n, T., AncĪ´n-Azpilicueta, C., 2007. Effect of SO2 on the formation and evolution of
volatile compounds in wines. Food Control 18, 1501–1506.

2184
 Mode of Inhibition of Į-Glucosidase and Į-Amylase by Polyphenol-Enriched Extracts of

Maqui (Aristotelia chilensis)

Francisca Acevedoa, Mónica Rubilara,b*, Barbara Palmaa, Carolina Shenea,b

a
Center of Food Biotechnology and Bioseparations, BIOREN, Universidad de La Frontera, Casilla 54-D,
Temuco, Chile (mrubilar@ufro.cl)
b
Technology and Processes Unit, CGNA, Universidad de La Frontera, Casilla 54-D, Temuco, Chile
(mrubilar@ufro.cl)

INTRODUCTION
Polyphenolic compounds are known to be useful in formulating of nutritional or medicinal
supplements for the treatment of several diseases. An important activity of polyphenols is the
inhibition of digestive enzymes, especially carbohydrate-hydrolyzing enzymes such as Į-amylase
and Į-glucosidase. Inhibitors of these enzymes are able to retard carbohydrate digestion, thus
causing a reduction in glucose absorption rate. Effective Į-amylase and Į-glucosidase polyphenol-
type inhibitors from natural resources have been reported to be useful in reducing postprandial
hyperglycemia [1]. To our knowledge, the inhibition mode of maqui leaf crude extract against Į-
glucosidase and Į-amylase has not been reported yet.

MATERIALS & METHODS

Leaves of maqui was collected from the Andes mountains in the Araucanía Region (Chile). The
collected material was dried at 35 °C, ground and sieved. The sample (5 g) was macerated with
ethanol (50% v/v in water, solvent-to-solid ratio of 5:1) at room temperature and filtered. Filtrate
was concentrated in a rotary evaporator at 35°C and lyophilized. The dry powder was dissolved in
ethanol 50% v/v. In order to examine the inhibition mode of maqui leaf crude extract, Į–amylase
and Į-glucosidase activities were measured with increasing concentrations of substrate in the
absence and presence of maqui leaf extract at different concentrations. Alpha-amylase activity was
quantified by measuring the maltose equivalents released from starch at 540 nm. Alpha-glucosidase
activity was quantified by measuring the para-nitrophenol equivalents released from pNPG at 400
nm. The Michaelis-Menten constant (Km), maximum enzyme reaction rate (Vmax) and the inhibition
mode of maqui leaf crude extract on the Į-amylase-catalyzed hydrolysis of starch and on the Į-
glucosidase-catalyzed reaction of pNPG were estimated using Lineweaver–Burk plots. In this
study, the initial velocity ‘v’ of the hydrolysis reactions catalyzed by Į- amylase was measured at
various substrate concentrations [S] (0.2 – 1% starch) in the absence and presence of several maqui
leaf crude extract concentrations [I] (10 – 100 ppm).

11th International Congress on Engineering and Food - Athens, Greece, 2011 2185
RESULTS AND DISCUSSION
Km value for Į-amylase was found to be 1.8% starch with a Vmax value of 6.8 μM min-1. The results
indicate that maqui leaf crude extracts act as mixed-type inhibitors, binding to either Į-amylase (E)
or enzyme-substrate (ES) complex, resulting in a decrease in the apparent affinity of Į-amylase for
starch (increase Km) and a decrease in the apparent Vmax. A similar type of Acarbose inhibition and
for the two Acarbose analogues for porcine pancreatic Į-amylases was reported by Yoon and
Robyt [5]. Similar behavior appears reported in literature for finger millet seed coat phenolics,
which behave as non-competitive inhibitors on pancreatic Į-amylase, being Km value for this
enzyme about 1% starch [3].
The v of the hydrolysis reactions catalyzed by Į-glucosidase was measured at various substrate
concentrations [S] (2-5 – 15-0 mM pNPG) in the absence and presence of several maqui leaf crude
extract concentrations [I] (0.05 – 5-0 ppm). The Km for Į-glucosidase was found to be 3.5 mM
starch with a Vmax value of 0.25 mM min-1. The results of this study indicate that binding of
phenolic compounds to enzyme affected the velocity of Į-glucosidase reaction rate, proportionally
to the concentration of the phenolic compounds in the reaction mixture, not modifying Km value.
Thus, a non-competitive inhibition by maqui leaf crude extracts upon Į-glucosidase-catalyzed
pNPG hydrolysis was found in this study. Reversible non-competitive inhibition of Į-glucosidase is
reported in literature for aqueous extracts from the gall of Rhus chinensis [2]. Alpha-glucosidase
inhibition by several flavonoids has been reported as mixed-type and almost non-competitive [4].

CONCLUSION
Furthermore, the evaluation of digestive enzyme inhibition in the extract represents a preliminary
approach to the potential biological properties of maqui leaves, which are used in traditional
medicine.

REFERENCES
[1] Gao H., Huang Y.N., Gao B., Xu P.Y, Inagaki C.; Kawabata J. 2008. Į-glucosidase inhibitory effect by the
flower buds of Tussilago farfara L. Food Chemistry, 106, 1195-1201.
[2] Shim Y.-J., Doo H.-K., Ahn S.-Y., Kim Y.-S., Seong J.-K., Park I.-S., Min B.-H. 2003 Inhibitory effect of
aqueous extract from the gall of Rhus chinensis on alpha-glucosidase activity and postprandial blood
glucose. Journal of Ethnopharmacology, 85, 283-287.
[3] Shobana, S.; Sreerama, Y.N., Malleshi, N.G. 2009. Composition and enzyme inhibitory properties of
finger millet (Eleusine coracana L.) seed coat phenolics: mode of inhibition of a-glucosidase and
pancreatic amylase. Food Chemistry 115, 1268–1273.
[4] Tadera K.; Minami Y.; Takamatsu K.; Matsuoka T. 2006.Inhibition of Į-glucosidase and Į-amylase by
flavonoids. Journal of Nutritional Science and Vitaminology. 52, 149-153.
[5] Yoon S.-H.; Robyt J.F. 2003Study of the inhibition of four alpha amylases by acarbose and its 4IV-a-
maltohexaosyl and 4IV-a-maltododecaosyl analogues. Carbohydrate Research, 338, 1969-1980.

2186
 Influence of pH variation during propolis extraction with the use of water as solvent

Beatriz C.B.S.Melloa, Paula M. Kakudab, Miriam D. Hubingera

a
Dept. of Food Engineering, Faculty of Food Engineering, University of Campinas, Brazil
(mhub@fea.unicamp.br)

INTRODUCTION
Propolis is a substance that shows a variable and complex chemical composition with
antimicrobial, antioxidant and antiviral properties, associated to its high concentration of
flavonoids and phenolic compounds. Due to these characteristics, which can bring health
benefits, propolis is considered a functional ingredient and has attracted much attention in
recent years as an important substance that can be used in foodstuffs, beverages, cosmetics and
medicine to improve health and prevent diseases. Considering the widespread use of propolis,
the objective of this work was to evaluate the effect of pH variation on propolis extraction,
prepared with water as solvent. Six different pHs were tested to each solvent and were
compared with samples without pH variation. Final extracts were quantified regarding
flavonoids and phenolic contents to verify the relation between the pH and solvent in the
extraction efficiency. Moreover, analyses of antimicrobial and antioxidant activities were
carried out for all the extracts.

MATERIALS & METHODS

Aqueous propolis extracts was prepared from crude propolis previously comminuted in a bench
blender, homogenized, weighed and mixed to deionized water (20% propolis and 80%
solvent). After five days at room temperature, in the dark, the sample was centrifuged at 8800g
for 20 min. Finally, the resulting extract was stored under refrigeration (4°C) in a closed
recipient in the dark. It was used HCl 1M and NaOH 1M to change the extraction pH.
Total flavonoid content of the propolis solutions was determined spectrophotometrically by the
aluminium complexation method [1]. The polyphenols in the propolis solutions were
determined by the Folin-Ciocalteau colorimetric method [2]. Antimicrobial activity of the
propolis samples was investigated by the disc diffusion method, using Staphylococcus aureus
[3]. Antioxidant activity was measured by FRAP and DPPH methods.

RESULTS & DISCUSSION

Table 1 shows the quantification of the extracted compounds for each sample and their
antioxidant activities. For the aqueous extracts, the results for flavonoid concentration showed
that all samples presented statistical differences compared to the initial solution (extracted
without pH variation) at 95% significance. However, only the samples with a more basic pH
(6.0 and 8.0) had an increase on concentration of the main functional compounds. This
behaviour was the same for phenolic compounds in the samples at pH 6.0 and 8.0, which had
statistical difference from the initial solution, at pH 4.3. Flavonoids and phenolic quantitative
results were compared in order to choose which combination of solvent and pH is the best one
to produce a novel propolis extract.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2187
 Table 1. Flavonoids, phenolic compounds and antioxidant activity in extracts
prepared with different pHs
Solution Flavonoids (mg/g)* Polyphenols (mg/g)** FRAP value a DPPH scavenging b
pH 2.0 12.08 ± 0.54 (a) 36.96 ± 0.04 (a) 42,44 ± 0,78 304,76 ± 7,41
pH 3.0 11.41 ± 1.56 (a) 35.48 ± 0.44 (b) 45,48 ± 1,33 385,71 ± 22,22
pH 4.3 23.67 ± 2.14 (d) 36.57 ± 0.35 (a.b) 24,17 ± 1,66 180,95 ± 7,41
pH 6.0 33.21 ± 2.62 (b) 40.37 ± 0.27 (c) 44,17 ± 0,57 190,48 ± 19,60
pH 8.0 61.42 ± 1.51 (c) 45.41 ± 0.40 (d) 44,44 ± 1,01 414,29 ± 46,26
Values are represented by mean ± S.D of three experiments
* quercetin equivalents
** gallic acid equivalents
a
In ȝmol Fe2+/mg dry weight of extract
b
(%Inhibition)
Means with different superscript letters within a column are significantly different at p< 0.05

Antimicrobial activity was higher to aqueous extract at pH 8.0 and ethanolic extract. Despite of
the increase on flavonoids and phenolic compounds in the basic aqueous samples, they did not
show the same activity against Staphylococcus aureus when compared to the ethanolic extract.
The aqueous sample without base addition and the sample at pH 6.0 had no activity against the
microorganism, while the sample at pH 8.0 showed a little activity, 62% lower than the activity
obtained for the ethanolic sample.

CONCLUSION
According to the results overview, the studied propolis extracts represent an important
functional product, rich in flavonoids and polyphenols. By using alkaline water as the
extraction solvent, the amount of extracted compounds increased as so their antimicrobial
activity comparing with the extract prepared without pH modification. In conclusion, water
could be used as an alternative solvent for propolis extraction, with a similar behaviour than the
ethanolic one and without its disadvantages.

REFERENCES
[1] Marcucci, M.C., Woisky, R.G. & Salatino, A. 1998. Use of aluminium chloride in the flavonoids
quantification of propolis samples. Mensagem Doce 46, 3-9. (In Portuguese)
[2] Kumazawa, S., Hamasak, T. & Nakayama, T. 2004. Antioxidant activity of propolis of various
geographic origins. Food Chemistry, 84, 329 – 339.
[3] Brindley Morgan, W.J. (1990). Guidelines for surveillance and control of antimicrobial resistances.
WHO/Zoonoses/90, 167, 15-17.

2188
 Modifier effects on Supercritical Fluid Extraction (SFE) of some Brazilian plants:

Antioxidant activity and Economical evaluation

Priscilla C. Veggi, Rodrigo N. Cavalcanti, M. Angela A. Meireles

LASEFI/DEA/FEA (School of Food Engineering)/UNICAMP (University of Campinas) – R. Monteiro

Lobato, 80; 13083-862, Campinas, SP, Brazil (meireles@fea.unicamp.br)

INTRODUCTION
Many Brazilian plants were found to be rich sources in antioxidant activities [1]. Pyrostegia
venusta (common names: flame vine, flaming trumpet, golden shower), Heteropterys
aphrodisiaca (nó-de-cachorro), Inga edulis (ingá-cipó), Hymenaea courbaril stilbocarpa
(jatobá) and Phaseolus vulgaris L. (beans) have been reported great natural antioxidant
properties. The important role of antioxidants in human health has been demonstrated, thus
increasing the interest in such products and their demand by consumers. Supercritical fluid
extraction (SFE) is a promising technology proved to obtain extracts with high quality by using
CO2. However, only CO2 is not sufficient to obtain the extracts with antioxidant properties due
to its low polarity. Then, the use of a modifier (or a co-solvent), can viabilize the extraction of
antioxidants via SFE. Concerning that, the use of simulation software to estimate the cost of
manufacturing (COM) of a product allows substantial cost, labor and time reduction in the
studying of industrial processes, making possible great capacity of analysis of the process.The
aim of this work is to evaluate the quality of the Brazilian plants extracts selected through the
antioxidant activity and perform the economical evaluation of the SFE process by estimating
the COM of the extracts.

MATERIALS & METHODS

To evaluate the effectiveness of SFE in obtaining extracts rich in antioxidant, the global yields
of these plants were determined at 323 K and 35 MPa by using CO2 and CO2 + ethanol (10%)
as a modifier. The antioxidant activity was determined by the linking capability of the free
radical DPPH (1,1-difenil-2-picrilidrazil). The software SuperPro designer 6.0® was used to
simulate the process and to estimate COM of extracts. The software considers the total capital
investment (FCI), raw materials (CRM), utilities (CUT), operating labor (COL) and so on. The
process was designs to run 7920-h per year, which corresponds to 330 days per year of
continuous 24-h per day operation. For the scale-up, the procedure assumed that the industrial
scale unit has the same performance as the laboratorial scale unit; this study considered setups
with extractor of 0.3m3.

RESULTS & DISCUSSION

Figure 1a, b show the results of global yields (% X0) and antioxidant activity (% SA) for SFE
with CO2 and CO2 + ethanol (EtOH). Antioxidant activities are expressed as the percentage of
potential antioxidant compounds present in the extract. Looking at Figure 1a it is possible to
notice that for SFE with CO2 the greatest amount of global yield was obtained for ingá-cipó
(IC) followed by jatobá (JB), nó-de-cachorro (NC), cipó-de-são-joão (CSJ), and bean (FJ). This

11th International Congress on Engineering and Food - Athens, Greece, 2011 2189
demonstrates that a higher recovery of extract is not always associated with a higher recovery
of antioxidant compounds. On the other hand, the highest value of antioxidant activity per
extract was achieved for jatobá followed by cipó-de-são-joão, bean, ingá-cipó, and nó-de-
cachorro. However, the highest antioxidant activity percentage was obtained for jatobá and had
the second best global yield as well.
In Figure 1b, it can be verified that the addition of ethanol as cosolvent in SFE process had a
positive influence in the extraction efficiency of all raw materials, the addition of ethanol as
modifier showed a great enhancement in antioxidant activity of SFE extracts for the samples
jatobá, ingá-cipó, and nó-de-cachorro; representing better antioxidant compounds sources
using SFE with ethanol than, using pure CO2, while bean and cipó-de-são-joão achieved better
antioxidant activity results by SFE with pure CO2.

(a) (b)

Figure 1. Global yields (X0) and antioxidant activities (%) of SFE extracts from cipó-de-são-joão (CSJ),
ingá-cipó (IC), nó-de-cachorro (NC), jatobá (JB), and bean (FJ) with CO2 (a) and CO2 + ethanol.

The specific costs obtained for the antioxidant compounds (COMAC) for SFE process using
CO2 + EtOH were a lot lower than those assessed for the process using only CO2. And also, the
COM was directly influenced by the global yield (X0) of extracts; in extractions with CO2 the
lower X0 of extracts lead to higher COM. the COM was directly influenced by the global yield
(X0) of extracts; in extractions with CO2 the lower X0 of extracts lead to higher COM.

CONCLUSION
The results indicated that all the global yields increased with the use of ethanol as co-solvent
instead of CO2 pure to obtain rich antioxidants extracts by SFE. Comparing all the extracts,
jatobá showed the highest antioxidant activity. According to the simulation process, all the
extracts obtained with the addition of co-solvent showed a lower COM.

REFERENCES
[1] Leal, P. F., Braga, M. E. M., Sato, D. N., Carvalho, J. E., Marques, M. O. M., & Merieles, M. A. A.
2003. Functional properties of spice extracts obtained via supercritical fluid extraction. J. Agric. Food
Chem. 51: 2520-2525.

2190
 Anthocyanin Extraction From Jabuticaba (Myrciaria cauliflora) Skins by Different

Techniques: Economical Evaluation

Priscilla C. Veggi, Diego T. Santos, M. Angela A. Meireles

LASEFI/DEA/FEA (School of Food Engineering)/UNICAMP (University of Campinas) – R. Monteiro

Lobato, 80; 13083-862, Campinas, SP, Brazil (meireles@fea.unicamp.br)

INTRODUCTION
Anthocyanins are a type of functional pigment responsible for a wide range of colors present in
vegetables, flowers, fruits, and derived products. It is known that anthocyanin pigments act as
strong antioxidants and are anti-inflammatory, with antimutagenic and cancer chemopreventive
activities [1].
Grape peels, grape by-products (constituted mainly by peels) and berries are well known for
their antioxidant properties due to the presence of anthocyanins and other phenolic compounds.
Many studies have been done to extract and evaluate these compounds on the industrial scale.
In Brazil another source seems promising; jabuticaba (Myrciaria cauliflora) is grape-like in
appearance and texture, although its skin is thicker and tougher. This fruit has a dark purple to
almost black skin color due to a high content of anthocyanins that cover a white gelatinous
flesh inside [2].
As the extraction procedure is of great importance for obtaining natural colorants, different
research groups have made an effort to develop an efficient extraction procedure. An efficient
extraction should maximize anthocyanin recovery with minimal degradation and result in an
extract with high antioxidant activity using environmentally friendly technologies and low-cost
raw materials. For this purpose different techniques were evaluated in terms of economical
feasibility for extraction of anthocyanins from skins of jabuticaba. Ultrasound assisted (UAE),
agitated bed (ABE), soxhlet and pressurized liquid extraction (PLE) methods were
economically compared. Ethanol was used as extraction solvent for all extraction techniques.
The simulation was conducted using the software SuperPro designer 6.0®.

MATERIALS & METHODS

Plant Material
Jabuticaba fruits (Myrciaria cauliflora) harvested from a plantation in the State of São Paulo,
Brazil, were acquired from a fruit and vegetable market center (CEASA-Campinas, Brazil).
Immediately after acquisition, the fruits were stored in the dark in a domestic freezer (-10ºC)
(Double Action, Metalfrio, São Paulo, Brazil) until sample preparation. Before extraction, the
fruits were manually peeled.
Economical Evaluation
The estimation of the cost of manufacturing (COM) was done for the crude extract obtained by
UAE, ABE, soxhlet and PLE. The main costs that compose the COM are similar to the ones
described by Turton et al. [3], which are given by total capital investment cost and operating
cost. The total capital investment cost represents the fixed capital investment (FCI), working
capital and start-up cost. The first one involves expenses with equipment, installation,

11th International Congress on Engineering and Food - Athens, Greece, 2011 2191
territorial taxes, engineering, etc., while the second one represents operating liquidity available
to a business, and finally, the start-up cost is associated with the beginning of operation and the
validation of the process. The operating cost represents direct costs that are directly dependent
on the production rate; it is composed of the cost of raw materials (CRM), the cost of the lost
solvent during the process, utilities cost (CUT), which represents the demand for steam and
cooling water required for the evaporator and condenser, electricity, and operational labor cost
(COL).

RESULTS & DISCUSSION

The COM obtained for the extract from jabuticaba skins extraction in UAE, ABE, soxhlet and
PLE processes for extractor capacities of 0.05, 0.1 and 0.3 m3 can be observed in Table 1.

Table 1. COM for extract´s global yield estimated for UAE, ABE, soxhlet and PLE
Extractor Global yield COM for Crude Extract
Extraction Technique
Capacity (m3) (%) (US$/kg)
UAE 0.05 794.46
0.10 11.93 530.22
0.30 401.21
ABE 0.05 1016.88
0.10 9.01 666.50
0.30 422.18
soxhlet 0.05 3020.00
0.10 9.92 1800.00
0.30 778.42
PLE 0.05 19.26
0.10 13.01 17.24
0.30 15.53

CONCLUSION
According to the results, PLE resulted in higher extraction efficiency followed by UAE,
soxhlet and ABE. PLE process also was the most economically viable method to obtain
extracts rich in anthocyanins due to the use of less solvent and time.

REFERENCES
[1] Kong, J., Chia, L., Goh, N., Chia, T., & Brouillard, R. 2003. Analysis and biological activities of
anthocyanins. Phytochemistry, 64(5), 923-933.
[2] Santos, D.T., & Meireles, M.A.A. 2009. Jabuticaba as a Source of Functional Pigments.
Pharmacognosy Reviews, 3(5), 127-132.
[3] Turton, R., Bailie, R.C., Whiting, W.B. & Shaeiwitz, J.A. 2003. Analysis, Synthesis and Design of
Chemical Process, Prentice Hall-PTR, New Jersey.

2192
 Study of cleaning efficiency of organic microfiltration membranes by attenuated total
reflectance infrared microspectroscopy

Tilahun K. Gelawa, Alexandre Trentina, Carme Güella, Montse Ferrandoa, Sílvia de Lamo-Castellvía,*

a
Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Païssos Catalans, 26 campus
Sescelades, 43007 Tarragona, Spain

INTRODUCTION
In membrane emulsification process, there are two different methods of operation, direct
membrane (DME) and premix membrane emulsification (PME). For DME, the to-be-dispersed
phase is pressed through a microporous membrane while the continuous phase flows along the
membrane surface. However, for the case of PME, a coarse premix is prepared and pushed
through a membrane producing finer droplets [1]. The main problem of PME is membrane
fouling, since both the continuous phase (with the emulsifier) and the disperse phase pass
through the membrane [2]. Fouling occurs due to the interaction between the membrane and
the components in the emulsion [3]. To reuse the membrane, it is necessary to clean it.
Membrane cleaning can be performed by a combination of water and air in either the forward
or background direction or applying chemical cleaning [4]. Moreover, it is also possible to
clean the membranes using chemicals like Tween 20 and Derquim + at specific concentration
and operating pressure. The objective of this work was to study the efficiency of different
cleaning protocols applied to reduce fouling phenomena produced during membrane
emulsification on nylon micromembrane by using infrared microspectroscopy (IRMS)
combined with multivariate analysis, specifically soft independent modelling of class analogy.

MATERIALS & METHODS

Oil/Water (O/W) emulsions were prepared using commercial sunflower oil (10% v/v, disperse
phase), MiliQ water (continuous phase) and whey protein solution (1% w/w) (WPC,
Lactalbumin® 75 L, from Milei - Stuttgart, Germany, emulsifier) using a two-step
emulsification process (premix emulsification procedure). After three emulsification cycles, the
membranes were cleaned using a combination of different concentration of Tween 20
(polyoxyethylene sorbitan monolaurate, from Sigma-Aldrich, Spain) and pressure of N2 on a
backflush mode. In this experiment five different cleaning protocols were tested by combining
different concentrations of Tween 20 (2, 3 and 4%) and N2 pressure (150 – 700 kPa). The
composition of different samples present on the nylon membrane were analyzed by ATR-FTIR
equipment (Illuminate IR, Smiths detection, 64 Clarendon Road, Watford, Herts WD17 1DA,
UK) interfaced with mercury-cadmium-telluride (MCT) photoconductive detector and
equipped with a microscope with a motorized x-y stage, 20x and 50x objectives, and slide-on
attenuated total reflection (ATR) diamond objective (Smiths detection, The Genesis Centre
Science Park South Birchwood Warrington, WA3 7BH, England). IRMS spectra of 10% (w/v)
whey protein solution, sunflower oil and Tween 20 were taken after the samples were placed
on the nylon membrane and dried. The spectrum was acquired from 800 to 4000 cm-1
wavelength with 4 cm-1 resolution. A background scan was taken before every sample. The
spectrum of each sample was obtained by taking the average of 128 scans to improve the
signal-to-noise ratio. Raw spectra were exported to Pirouette multivariate analysis software for
soft independent modeling of class analogy (SIMCA) analysis.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2193
RESULTS & DISCUSSION
Spectra of new and fouled membranes were analyzed by SIMCA.The infrared spectra analysis
(1800-900 cm-1) using SIMCA classification models of new and fouled membranes, sunflower
oil and 10% whey protein solution, permitted tight clustering, clear differentiation and zero
misclassifications among samples. Discriminating power of SIMCA of new and fouled
membranes, sunflower oil and 10% whey protein solution, showed four strong spectral bands
at 1550, 1516, 1099 and 1057 cm-1 (data not shown). The discrimination power is a measure of
variable importance in infrared frequency and contributes to the development of the
classification models [5]. The first two IR bands at 1550 and 1516 cm-1, were associated to N-
H bending and carbonyl stretching bands, respectively [5]. These two bands are diagnostic of
secondary amides (whey protein and nylon membrane). The last two IR bands, 1099 and 1057
cm-1, were related with asymmetric and symmetric stretching modes of C-O-C
(polysaccharides) of esters presents in sunflower oil and whey protein. These four bands
clearly differentiate between fouled and new membrane spectra. Moreover, the interclass
distance (ICD) of the SIMCA classification model ranged from 6.1 to 39.7 (data not shown)
showing chemical differences among the membrane samples. In the other hand, the infrared
spectra analysis (1800-900 cm-1) of different clean membrane samples showed tight clustering
and clear differentiation among the different cleaning protocols applied and fouled membrane
(data not shown). From the ICD data, the membranes cleaned with 4 or 3 % Tween 20
combined with 500 kPa or 700 kPa of nitrogen pressure had the highest values, 9.7 and 9.6
respectively. The discriminating power of SIMCA classification model of fouled and clean
membranes showed a band at 1629 cm-1 linked to the presence of protein from whey protein or
the nylon membrane.

CONCLUSION
ATR-IRMS combined with multivariate analysis is a valuable tool to obtain information about
membrane surface. It was simple and easy to differentiate between membrane samples (new,
fouled and cleaned) and also to detect the most effective membrane cleaning protocols among
those tested in this experiment. IRMS combined with multivariate data analysis has not been
used for membrane characterization and with this research we show that could be an efficient
and rapid technique.

REFERENCES
[1] Suzuki K., Fujiki I. and Hagura Y.1998. Preparation of corn oil/water and water/corn oil emulsions
using PTFE membranes. Food Science and Technology International, Tokyo 4 (2), 164-167.
[2] Trentin A., Güell C., López F. and Ferrando M. 2010. Microfiltration membranes to produce BSA-
stabilized O/W emulsions by premix membrane emulsification. Journal Membrane Science 356.
[3] Belfort G., Davis R.H. and Zydney A.L. 1994. The behavior of suspensions and macromolecular
solutions in crossflow microfiltration. Journal Membrane Science 96, 1–58.
[4] Kuzmenk D., Arkhangelsky E., Belfer S., Freger V. and Gitis V. 2005. Chemical cleaning of UF
membranes fouled by BSA. Desalination 179, 323-333.
[5] Helm D., Labischinski H. and Naumann D. 1991. Elaboration of a procedure for identification of
bacteria using Fourier-Transform IR spectral libraries: a stepwise correlation approach. Journal of
Microbiological Methods 14, 127-142.

2194
Comparative study on quality evaluation of buffalo meat slices incorporated with finger millet, oats

and chickpea

Mahjabeen Siddiquia, Mohammad Ali Khanb

a
Aligarh Muslim University, Aligarh, India (mahjabin111@yahoo.co.in)
b
Aligarh Muslim University, Aligarh, India (makamu4@gmail.com)

INTRODUCTION
A buffalo meat is a major and cheapest source of protein in India, especially for those consuming a non-
vegetarian diet. It is also a major source of healthy iron in the Indian diet. [1]. Meat and meat products are
essential for a balanced diet, although it must also be remembered that they are susceptible to
modifications to give them a “healthier” appearance. Other non meat additives used as fillers/binders
include wheat flour in chicken nuggets [2], texturized soy protein in chicken kebab [3], cowpea and
peanut flour in chicken nuggets [4], Bengal gram flour and maida in chicken patties, and green and black
gram flours in buffalo meat burger. Ragi or finger millet (Eleusine coracana) is a low-cost cereal and is a
rich source of calcium, iron and phosphorous. Chickpeas are high in protein and a helpful source of zinc
and folate. Oats (Avena sativa L.) is a typical cereal containing ß-glucans, which have an effect on blood
cholesterol levels and control of lipoprotein metabolism [5]. The object of this paper is to evaluate the
effect of adding functional ingredients such as chickpea, finger millet and oats on the physicochemical,
microbiological and sensory characteristics of buffalo meat slices.

MATERIALS & METHODS

The preparation of buffalo meat slices incorporated with chickpea, finger millet and oats are the same and
the composition selection of all the ingredients are mentioned in Table 1 on the basis of preliminary
organoleptic trails. These ingredients were weighed and mixed properly, wet-ground and prepared in the
form of dough, which was filled in stainless steel brick shaped moulds.

Table1:- Ingredients of buffalo meat slices incorporated with chickpea, finger millet and oats

Ingredients Control Chickpeas based Finger millet Oats based

S.No
Quantity (gm/kg)
1 Buffalo lean meat 540g 540g 540g
2 Chickpea 400g -- --
3 Finger millet -- 400g --
4 Oats -- -- 400g
5 Big cardamom 4g 4g 4g
6 Cinnamon 3g 3g 3g
7 Black pepper 3g 3g 3g
8 Garlic paste 5g 5g 5g
9 Ginger paste 5g 5g 5g
10 Turmeric powder 6g 6g 6g
11 Red chilly powder 14g 14g 14g
12 Table salt 20g 20g 20g
Note: not added (--)

11th International Congress on Engineering and Food - Athens, Greece, 2011 2195
RESULTS & DISCUSSION
Variation of pH, Moisture content, TBA number, ash, protein, fat, TPC and Yeast and mould count of
chickpea, finger millet and oats incorporated buffalo meat slices during storage have been shown Figures
1 to 8.

CONCLUSION
Based on the above results, the following conclusions can be drawn: A control buffalo meat slice which
were prepared without fortification of any cereal or millet flour. The fortification with finger millet, oats
and chickpea in the development of buffalo meat slices could impart multi-pronged beneficial attributes
together with a host of beneficial physiological effects. Keeping in view the many promising health
effects, such food adjuncts could be regarded as ‘neutraceuticals’, which makes the food healthier.

Thus, among the processed meat slices with fortification have the advantage of a longer shelf life (120
days) and low cost of processing. The longer shelf life of these products at refrigerated temperature and
good nutritive/medicinal values may add great convenience to many meat consumers. Finger millet, oats
and chickpea incorporated samples are lower in fat and higher in protein content and ash content, which
indicates that the finger millet, oats and chickpea incorporated samples are rich in mineral source in
comparison to control samples.

REFERENCES
[1] Fernández-Ginés JM, Fernández-López J, Sayas-Barberá E, & Pérez-Alvarez Ja. 2005. Meat Products as
Functional Foods: A Review—Journal of Food Science, 70(2), 37-43. [2] Rao K.H., Singh R.P., Anjaneyulu A.S.R.,
Rao K.V.S.S. & Yadav P.L. 1997. Effects of caseinases and refined wheat flour on the quality of chicken nuggets from
spent hens. Ind. J. Anim. Sci., 67, 1004–1006. [3] Mir Salahuddin, Kondaiah N. & Anjaneyulu A.S.R. 1991. Effect of
maida, potato and texturized soya protein as binders on the quality of chicken and mutton kababs. J. Food Sci.
Technol., 28, 301–303. [4].Prinyawiwatkul W., Mcwatters K.H., Beuchat L.R. & Phillips R.D. 1997. Physicochemical
and sensory properties of chicken nuggets extended with fermented cowpea and peanut flours. J. Agric. Food Chem.,
45, 1891–1899. [5] Braaten J. T., Wood P. J., Scott F. W., Wolynetz M. S., Lowe M. K., Bradley-White P., & Collins
M. W. 1994. Oats ß-glucan reduces blood cholesterol concentration in hypercholesterolemic subjects. Eur. J. Clin.
Nutr., 48, 465.

2196
 Microencapsulation of tocopherols in lipid matrix by spray chilling method

Oscar Diaz Gamboa, Aparecida Lireny Guaraldo Gonçalves, Raymundo Carlos Grosso

Faculty of Food Engineering, University of Campinas, Campinas, Brazil

(oscarw07@fea.unicamp.br)

INTRODUCTION

Vitamins are essential micronutrients that contribute to normal growth and maintenance of health, however, vitamin E is
very unstable because it is slowly oxidized by atmospheric oxygen. Microencapsulation is a technology for coating
substances for protection and/or controlled release of them. Likewise, microencapsulation using lipids have great flexibility
regarding packaging material and size of the particles.This project aimed to evaluate the stability of the microcapsules of Į-
tocopherol in different storage conditions of temperature in a period of time.

MATERIALS & METHODS

Raw Material. Interesterified fat (cottonseed oil and fully hydrogenated palm oil), fully hydrogenated soybean oil (OSTH),
and as active ingredient to be encapsulated Į-tocopherol. To obtain microcapsules of the lipid matrices were melted at a
temperature of 65 º C. The Į-tocopherol was added followed by homogenization in ultra-Turrax for 5 min. The lipid
microparticles suffered in spray atomizer heated fluid also double to 65 ° C and air pressure of 0.25 MPa, with atomization
performed inside a chamber cooled to 10°C. Four trials were prepared, four trials coded A, B, C, D respectively, which were
submitted by 180 days storage at three different temperatures (BOD at 22°C and -18°C freezer, in the absence of light and
temperature of 25 ± 3°C with light). Determination of encapsulation efficiency. Performed according to AOCS method Ce
8-89 [1]. Thermal Analysis.Performed according to AOCS method Cj 1-94, 2004 [2]. X-ray diffraction Polymorphic forms
of the fat on the microcapsules were analyzed according to AOCS Method Cj 2-95, 2004 [2].The Index of Relative
Crystallinity (Ic) was quantitatively estimated according to the method proposed by Rabek (1980) [3].

RESULTS & DISCUSSION

Encapsulation efficiency values determined in tests, one day after production were above 90%. Likewise the values for the
retention of Į-tocopherol (Table 1) of the microcapsules after 180 days of storage at three temperatures were considerably
higher for the systems studied ranged from 99.7 to 94.1%. The small variation observed over time in each test individually
assessed indicates that the lipid matrix using interesterified fat and FHSO, may have functioned as agents for preventing the
expulsion of the core material over time and creating a good accommodation microstructure for the drug.
Thermal behavior. The results of thermal properties of melting and crystallization of lipid microcapsules obtained by
Differential Scanning Calorimetry (DSC) at time zero was evaluated with the objective of verify possible differences in
relation to different proportions of lipid / core material because it could have different crystallization and melting behaviors
for obtained peaks. The values obtained for the peaks recorded in both the curve of melting and crystallization indicated as
being directly related to the proportion of lipid / core material used for the production of microcapsules. The melting
temperature "endset" occurred over a range of 62.2 °C to 64.0 °C corresponding to the tests B and C respectively, which
showed lower and higher amount of lipid matrix. Similarly the values of crystallization temperature curve indicate also be
related to the amou lipid in the matrix, because these values varied over a range of -2.5 °C to -7.5 °C corresponding to the
tests C and B respectively with higher and lower amounts of lipid matrix present in the microcapsules. So even if these
effects also showed an association with the respective reduction and increase in the values of melting and crystallization
enthalpy (ȴH). The melting curve was characterized by the formation of two distinct endothermic peaks separated by an

11th International Congress on Engineering and Food - Athens, Greece, 2011 2197
exothermic peak could be attributed to the polymorphism of FHSO. For crystallization curves the thermograms obtained
were characterized by the formation of two peaks of a smaller magnitude that appears close to 0 ° C and another large peak
near 40 ° C represents the crystallization of FHSO. Regarding the presence of Į-tocopherol on the microcapsules, the results
indicate that it did not affect the melting and crystallization of microcapsules in the tests, due to solubility that exists in the
lipid matrix.

Table 1. Retention values of tocopherol in the microcapsules after 180 days.

Temperature of storage Trials % Retention SD
A 99.0 ± 0.42 to 0.45
22°C (BOD) B 99.0 ± 3.89 a 4.20
C 98.5 ± 7.35 to 7.16
D 99.1 ± 3.25 to 5,04
A 94.8 ± 2.05 to 2,21
25 ± 3 °C (Environment) B 94.1 ± 0.283 a 0,34
C 98.8 ± 2.55 a 2,90
D 94.8 ± 1.28 to 1,34
A 99.4 ± 1.41 to 0,46
-18 ° C (Frezzer) B 99.7 ± 4.17 to 1,27
C 99.1 ± 6.50 a 7,32
D 99.7 ± 0.78 to 0,80
A: (ratio of lipid / core material 90/10), B: (ratio of lipid / core material 80/20), C: (ratio of lipid / core material 95 / 5), D:
(ratio of lipid / core material 85/15).

X-ray Diffraction. Diffractograms of the test capsules stored for 180 days at different temperatures showed similar specter
and shpae of the obtained curves.The results demonstrated the presence of three major peaks detected in the following
angles 2ș = 19.3 ° d = 4.6A, 2ș = 22.8 ° d = 3.8A and 2ș = 23.1 ° d = 3.7A. The standards were similar and literature
usually associate it with the polymorphic form ȕ, characteristic of most triacylglycerols and fatty acids.While the
percentages of crystallinity of the trials of lipid microcapsules were relatively lower than 30% without significant difference
during storage time. The low crystallinity observed for the formulations have been driven by the presence of amorphous
solids which are lipid but not crystalline that contribute to increase the retention of core material inside the matrix. Although
there are no quantitative data in the literature, the benefits of amorphous structures on microencapsulation efficiency of
products are well known.

CONCLUSION

The production of lipid microparticles containing Į-tocopherol is possible using lipid matrices using spray chilling with
good efficiency and high levels of retention of the active product, ranging from 94.1 to 99.7%.The lipid microparticles
showed good stability over time and temperature.

REFERENCES
[1] AOCS 1989.American Oil Chemists' Society. Official methods and recommended practices of the AOCS. 4th. ed. Champaign.
[2] AOCS 2004.American Oil Chemists' Society. Official methods and recommended practices of the AOCS. 4th. ed. Champaign.
[3] Rabek, JF 1980.Experimental Methods in Polymer Chemistry: Applications of Wide-Angle X-Ray Difraction (WAXD) to the
Study of the Structure of Polymers, Wiley-Interscience Chichester, UK p. 505.

2198
 Amino acid profile of Sous vide cooked poultry breast meat products

Kristine Ramanea, Ruta Galoburdaa, Viesturs Kreicbergsa, Ilona Vanagab

a
Latvia University of Agriculture, Faculty of Food Technology, Jelgava, Latvia
e-mail: ramane.kristine@gmail.com
b
Research Institute of Biotechnology and Veterinary Medicine “Sigra”,
Latvia University of Agriculture, Sigulda, Latvia

INTRODUCTION
Amino acid composition greatly determines the nutritive value of meat product. Chicken meat
is characterized by a high content of lysine, leucine, aspartic acid and glutamic acid [1]. Creed
& Reeve described that Sous vide cooking reduces heat damage to proteins, diminishes the loss
of liquids and aroma compounds at the same time providing longer shelf life comparing to
traditional cooking methods [2]. The improvement of tenderness in meats is mainly caused by
changes in structure of connective tissues solubilised by heat, while at the same time heat-
denaturation of myofiblrillar proteins generally causes meats toughening. In order to enhance
nutritional value of the meat product – vegetable-and-fruit additive can be used [3]. When the
transverse shrinkage to the fibre axis occurs mainly at 40–60 ºC this widens the gap already
present at rigor between the fibres and their surrounding endomysium. At 60–70 ºC the
connective tissue network and the muscle fibres co-operatively shrink longitudinally. It is then
presumed that water is expelled by the pressure exerted by the shrinking connective tissue on
the aqueous solution in the extracellular void [4]. As a result amino acid composition in cooked
meat is changed compared to raw product. The aim of this research was to evaluate amino acid
profile of four marinated, sous vide cooked poultry meat products made from broiler’s fillet
(A); broiler’s fillet with vegetable-and-fruit additive (B); mature hen’s fillet (C); mature hen’s
fillet with vegetable-and-fruit additive (D).

MATERIALS & METHODS

Carcasses of slaughtered broilers of the cross Ross 308 and parents’ stock hens after reaching
rigor mortis were randomly selected for separating a fillet (musculus pectoralis). The obtained
skinless fillets together with other ingredients were packaged in polyamide/polyethylene
(PA/PE) pouches (film thickness 90 μm, pouch size 230×145 mm), vacuum sealed, marinated
and sous vide cooked according to the technology described in the patent of Republic of Latvia
no. 14095 [3]. The amino acid profile was determined using HPLC-MS by method LVS ISO
13903:2005. Amino acid content is reported as average of three replications in g 100 g-1 dry
weight (DW).

RESULTS & DISCUSSION

Chemical composition showed significant differences (p<0.05) between broiler and hen meat,
regarding water and protein content. Mature hen’s fillet has lower water content and higher
protein content. Moreover amino acid content and especially content of individual essential
amino acids (Table 1) varies with bird age, being lower in mature hen’s fillet compared to
broiler’s fillet. In the thermal treatment process a cooking loss was observed due to loss of
water, fat, and soluble proteins. However relatively higher loss of water was observed thus the
total proportion of proteins per 100 g of product was increased after heat treatment.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2199
 Table 1. Content of essential amino acids in broiler’s fillet, hen’s fillet, and their products, g 100 g-1 DW
Broiler’s fillet Mature hen’s fillet
Chilled A B Chilled C D
Total amino
acids 52.58±0.42 50.82±0.26 44.78±0.51 49.43±0.12 44.56±0.25 43.94±0.25
Incl. essential
amino acids
Phenylalanine 3.01±0.29 2.93±0.09 2.63±0.01 3.20±0.03 2.92±0.12 2.97±0.19
Isoleucine 2.17±0.18 2.03±0.07 1.83±0.02 2.22±0.04 2.16±0.12 2.13±0.18
Leucine 4.33±0.36 4.05±0.15 3.65±0.04 4.52±0.02 4.25±0.23 4.14±0.36
Lysine 2.76±0.01 2.72±0.05 2.57±0.02 1.80±0.01 1.76±0.12 1.79±0.18
Methionine 2.53±0.13 2.32±0.05 2.11±0.34 1.51±0.02 1.41±0.03 1.22±0.05
Threonine 2.92±0.28 2.73±0.02 2.52±0.04 2.58±0.02 2.54±0.15 2.44±0.20
Valine 4.04±0.42 4.00±0.10 3.54±0.19 3.03±0.03 2.72±0.09 2.63±0.20
The highest content of total amino acids among studied samples was detected in raw broiler
fillet 52.16 g 100 g-1. It was reduced to 50.82 g 100 g-1 after heat treatment or to 44.78 g 100 g-1
when it was heat treated together with vegetable-and-fruit additive. Similar loss of amino acids
was observed in hen’s fillet after cooking, but it was not significantly different in both
thermally treated products (without/with vegetable-and-fruit additive (p>0.05). Among
essential amino acids the highest relative decrease is observed for methionine in thermally
treated broiler’s fillet either without or with additive, as well as in hen’s fillet with additive. It
can be related to the high activity of sulfide group of this amino acid, for example, it is readily
oxidized into the sulfoxide and further in the sulfone.

CONCLUSION
In cross Ross 308 broiler’s fillet significantly higher content of essential amino acids –
methionine, lysine, and valine was observed comparing to parent’s stock hen’s fillet of the
same breed. Relative decrease in essential and non-essential amino acid content of broiler’s
fillet product sous vide cooked together with vegetable-and-fruit additive is more than two
times higher comparing to relative decrease in the same product cooked without additive. In
thermal treatment process the highest relative decrease among studied amino acids was
observed for methionine.

REFERENCES
[1] Sales J. & Hayes J. P. 1996. Proximate, amino acid and mineral composition of ostrich meat. Food
Chemictry, 56(2), 167-170.
[2] Creed P. G. & Reeve W. 1998. Principles and applications of sous vide processed foods, In: Ghazala
G. (Ed.) Sous vide and cook-chill processing for food industry. Aspen Publishers Inc, Gaithersburg,
Maryland, 25-56.
[3] Dukalska L., Ramane K., Galoburda R. & Seglina D. 2010. Process to prepare chicken fillet with
vegetable and fruit said dish by Sous vide packaging. The Official Gazette of the Patent Office of the
Republic of Latvia - "Patenti un precu zimes", 20 March, 2010, pp. 431.–432.
[4] Tornberg E. 2005. Effects of heat on meat proteins – Implications on structure and quality of meat
products. Meat Science, 70, 493-508.

2200
 Antioxidant activity and porphyran content in hydrothermal extracts of Porphyra
Yezoensis (Susabinori)

Chinami Goto a, Siti Machmudahb, Mitsuru Sasakia, Motonobu Gotob,

Kiyoka Okaic, Yasuji Okaid, Shoji Kondoe

a
Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan
(106d8205@st.kumamoto-u.ac.jp)
b
Bioelectrics Research Center, Kumamoto University, Kumamoto, Japan
c
Department of Food and Nutritional Environment, Kinjo Gakuin University, Nagoya, Japan
d
Department of Human Life Science, Osaka Kun-Ei Women's College, Setsu, Japan

INTRODUCTION
There is a mechanism that produces active oxygen in the human body. The active oxygen
damages the cell function and causes many chronic diseases such as cancer, diabetes and
arteriosclerosis. At the same time, various defense functions exist in our human body that
eliminate and control against the production of the active oxygen. However, since these
functions decrease with age it is thought to be important to take foods which contain
antioxidation effect to prevent from the chronic diseases. Porphyra yezoensis (Susabinori),
which is a kind of seaweed marine biomass, has been eaten since early times in Japan.
Nonetheless, most of the remaining after the harvest of the seaweed is sent to the waste.
Sulfated polysaccharide such as Porphyran (POR), antioxidant activity substance, fat and
proteins are contained in seaweed contain. These components in Porphyra yezoensis are
thought to be produce antioxidant activity. It is useful to develop a technology to produce
valuable materials from unused part of marine biomass.
The aim of this study was set to determine the effective extraction method of useful
components from Porphyra yezoensis in order to use the seaweed effectively [1, 2]. Thus, POR
and others are extracted from the seaweed using subcritical water and the effects of extraction
temperature and time on extraction rate was investigated as well as molecular weight
distribution and measuring the antioxidant activity in the extract.

MATERIALS & METHODS

A sample was made by reducing the seaweed (Porphyra yezoensis without heat treatment) to
powder with particles less than 3 mm diameter using a blender. 0.1 g of seaweed powder and 5
ml of distilled water were placed into a batch extractor with volume of 8.8 ml and seal after
replacing the inside with argon gas. When the heater of the extractor reached the desired
temperature, place the reactor and start mixing. The experimental conditions of extraction were
temperature 140-220 oC and time 35-200 min. Then, immediately cool down the reactor with
water after the reaction and separate the extract and the residue using a centrifuge.
Moreover, the measurement of antioxidant activity was taken using DPPH method where each
extract was measured. Also, TOF-MS was used to analyze the molecular weight distribution.

RESULTS & DISCUSSION

Antioxidant material is affected by the extraction temperature and the time, and the extract
shows a higher antioxidant activity with temperature more than 200oC. Comparing with the

11th International Congress on Engineering and Food - Athens, Greece, 2011 2201
extracts extracted below 180oC, the temperatures above 200oC has higher activity at any
extraction time and shows a drastic increase in activity. However, the temperature condition
below 200oC also shows an increase in the antioxidant activity with the extraction temperature.
From the TOF-MS results, it was found that the smaller molecular weight components increase
with a higher extraction temperature and a longer extraction time. Furthermore, the higher the
extraction temperature, more the extracts with smaller molecular weight components were
obtained in a shorter time. Consequently, the results of antioxidant activity and molecular
weight show that the antioxidant activity increases with the extracts with smaller molecular
weight.
(a) (b)
㻞㻡㻚㻜㻜 㻡㻚㻜㻜

㻞㻜㻚㻜㻜 㻠㻚㻜㻜

㻟㻘㻢㻙㻭 䡊 㻳 㼍 㼘㼇㼙 㼓㼉
㻤㻜䉝
㻟㻚㻜㻜 㻤㻜䉝
㻰 㻙 㻳 㼍 㼘㼇㼙 㼓㼉

㻝㻡㻚㻜㻜
㻝㻞㻜䉝 㻝㻞㻜䉝
㻝㻜㻚㻜㻜 㻝㻢㻜䉝 㻞㻚㻜㻜 㻝㻢㻜䉝
㻡㻚㻜㻜 㻝㻚㻜㻜

㻜㻚㻜㻜 㻜㻚㻜㻜
㻜 㻡㻜 㻝㻜㻜 㻝㻡㻜 㻞㻜㻜 㻜 㻡㻜 㻝㻜㻜 㻝㻡㻜 㻞㻜㻜
㼀㼕㼙㼑㼇㼙㼕㼚㼉 㼀㼕㼙㼑㼇㼙㼕㼚㼉

Figure 1 A yield of the sugar of every extraction time at each temperature

(a)D-galactose (b)3,6-anhydrogalactose

㻡㻜㻚㻜
㻭㼚㼠㼕㼛㼤㼕㼐㼍㼠㼕㼢㼑㻌㼍㼏㼠㼕㼢㼕㼠㼥㼇㻑㼉

㻠㻜㻚㻜
㻟㻜㻚㻜 㻤㻜䉝
㻝㻞㻜䉝
㻞㻜㻚㻜 㻝㻢㻜䉝
㻝㻜㻚㻜
㻜㻚㻜
㻜 㻡㻜 㻝㻜㻜 㻝㻡㻜 㻞㻜㻜
㼀㼕㼙㼑㼇㼙㼕㼚㼉

Figure 2 Antioxidant activity of the extracts

CONCLUSION
By subcritical water treatment, POR was successfully recovered in high yield, where the
optimal temperature was 160 oC. The molecular weight of POR depends on the temperature
and time. The highest antioxidant activity was obtained at 160 oC.

REFERENCES
[1] Goto C., Ooga S., Izumi J., Machmudah S., Sasaki M., Goto M., Okai K., Okai Y. & Kondo S. 2010.
Antioxidant activity and porphyran content in the extracts of Porphyra yezoensis (Susabinori) using
hydrothermal treatment. Pacifichem, Honolulu.
[2] Oga S., Tanaka M., Kondou S., Sasaki M. & Goto M. 2009. Characterization of porphyran extracted
from Porphyra yezoensis (Nori) using subcritical water, Proc. Supergreen, Sendai, 2P-308.

2202
 Effect of frozen storage on the quality of camu camu (Myrciaria dubia (H. B.K.)

McVaugh,) pulp

Souza, A.L.R.a, Pagani, M.M.b, Gomes, F. S.b, Cabral, L.M.C.b

a
Department of Food Science and Technology/ UFRRJ, Rio de Janeiro-RJ, Brazil (hoglan@bol.com.br)
b
Embrapa Food Technolog,. Rio de Janeiro-RJ, Brazil (lcabral@ctaa.embrapa.br)

INTRODUCTION
Camu camu (Myrciaria dubia (H. B.K.) McVaugh, Myrtaceae) fruit has a high economical
potential as a natural source of vitamin C. Its pulp has the vitamin C content ranging from 1000
to 3000mg/100g [1], higher than the all citrus and acerola fruit, considered one of the main
sources of vitamin C. Moreover, the recent interest of polyphenols in plants has focused on its
great potential to benefit human health, with special reference to polyphenols in fruits and
vegetables [2]. Fresh fruits and vegetables, when harvested, continue to undergo chemical
changes which can cause spoilage and deterioration of the product. Freezing is a simple, quick
way to preserve foods. Fruits freeze well and retain their distinct fruit flavor. Texture is usually
softened somewhat by freezing, but serving partially frozen fruit with ice crystals will
compensate for texture changes. Lighter colored fruits may require special treatment to retain
color, texture and flavor. The processing of camu camu as frozen pulp is an alternative for its
conservation, increasing its shelf life, while maintaining their nutritional characteristics [3].
Thus, the objective of this work was to evaluate the effect of freezing on the functional quality
of the camu camu pulp.

MATERIALS & METHODS

Camu-camu fruits were obtained at the experimental fields of Embrapa Western Amazonia,
located at Manaus - Amazonas state, Brazil. The fruits were frozen and transported to Rio de
Janeiro where they were extracted using a pulper machine with a 0.6 mm diameter sieve. The
obtained pulp was homogenized and stored in plastic packing’s of 0.5kg at -18°C for 15
months. Samples were analyzed regarding acidity, pH, soluble and total solids, anthocyanins
and antioxidant capacity [4, 5, 6, 7, 8].

RESULTS & DISCUSSION

It was observed that the vitamin C content remained quite high, as cited by Rodrigues et al. [9]
and Rufino et al. [7].
The different vitamin C content in the camu-camu is a function of its maturity stage. The
vitamin C content found for the frozen camu camu pulp storage for 15 months was higher than
many fruits known for their high content of this vitamin like acerola and citrus.
Regarding the phenolic concentration and antioxidant activity, the values were also within the
expected range, indicating that the frozen camu camu pulp had a high level of phenolic
compounds and antioxidant activity even after 15 months storage. Barreto [10] determined the
acidity ranging from 1.94 to 2.07 g citric acid /100g, from 2.87 to 2.90 for pH and 5.30 to
5.75°Brix to the soluble solids content.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2203
 Table 1. Characterization of camu-camu pulp1

1
Mean ± Standard Deviation

CONCLUSION
Storage at -18 ° C retained the main functional characteristics of the camu camu pulp even
after a period of 15 months, suggesting the efficiency of freezing on the preservation of the
main physicochemical characteristics of this fruit.

REFERENCES
[1] Rodrigues, R. B., Marx, F. 2006. Camu camu [Myrciaria dúbia (H. B. K.) Mc Vaugh]: a promising
fruit from the Amazon Basin. Nutrition, 30 (9), 376-381.
[2] Clemente, C. R., da Silva, D. F. 1994. Amazonian small fruits with commercial potential. Fruit Var.
J., 48, 152-158.
[3] Rodrigues, R.B., Menezes, H.C., Cabral, L.M.C, Dornier,M., Rios, G.M., Reynes, M. 2004. Evalution
of reverse osmosis and osmotic evaporation to concentrate camu-camu juice (Myrcyaria dubia). Journal
of Food Engineering, 63, 97-102.
[4] AOAC. American Official of Analytical Chemists. Official methods of analysis of AOAC
International. 17. ed. Washington, 1997.
[5] Wrolstad, R. E. & Giusti, M. M. Characterization and mesasurement of anthocyanins by UV-visible
spectroscopy. In WROLSTAD, R. E. (Ed.). Current Protocols in Food Analytical Chemistry. New York:
Wiley, 2001.
[6] Re, R.; Pellegrini, N.; Proteggente, A. Pannala, A.; Yang, M.; Rice-Evans, C. Antioxidant acitivity
applting an improved ABTS radical cation decolorization assay. Free Radical Biology.1999.
[7] Rufino, M.S.M., Alves, R.E., Brito, E.S., Jiménez, J.P., Calixto, F.S., Filho, J.M. 2010. Bioactive
compounds and oxidant capacities of 18 non-traditional tropical fruits from Brazil. Food chemistry. 121,
996-1002.
[8] Georgé, S., Brat, P, Alter, P., AMIOT, M.J. 2005 Rapid determination of polyphenols and vitamin C
in plant-derived products Journal of Agricultural and Food Chemistry, 53, 1370-1373.
[9] Rodrigues, R. B., Menezes, H. C., Cabral, L. M. C., Dornier, M., Rios, G. M., Reynes, M. 2001
Amazonian fruit with a high potential as a natural source of vitamin C: the camu-camu (Myrciaria dúbia).
Fruit, 56, 345-354.
[10] Barreto, A. G., Carvalho, R. A., Cabral, L. M. C., Matta, V. M. , Freitas, S. P. 2009. Concentration
by reverse osmosis of camu camu juice. In: 3rd International Simposium on Huaman Health Effects of
Fruits and Vegetables. 1, 254-254.

2204
 Effect of semolina particle size on the cooking kinetics and quality of spaghetti

Giampiero Sacchettia, Giuseppe Coccob, Domenico Coccob, Lilia Neria, Dino Mastrocolaa

a
Department of Food Science, University of Teramo, Italy (gsacchetti@unite.it)
b
Pastificio Cav. Giuseppe Cocco, Fara S. Martino, Chieti, Italy (info@pastacocco.com)

INTRODUCTION
Semolina particle size is a key factor in pasta making and fine semolina is preferred by the
pasta industry since it gives a higher yield upon milling and shows a high hydration rate, thus
facilitating the mixing and further extrusion process.
However the reduction process required for fine semolina making could determine adverse
effects on quality due to starch damage such as: a higher reducing sugar content, lower starch
gelatinization temperature and solute leakage from the starch granule, resulting in higher pasta
stickiness and tendency to clump. Starch damage is more likely to occur when hard wheat
(Triticum durum, Desf.) varieties with hard kernels are used for pasta production.
This work was aimed to study the effect of semolina particle size on the chemical composition
and cooking quality of spaghetti.

MATERIALS & METHODS

A hard wheat (Triticum durum, var. Kronos) with high hardness value (162 N) was milled in
order to reach an average particle size of 275, 375 and 475 ȝm corresponding to medium,
medium coarse and coarse semolina respectively.
Semolina particle-size distribution was determined with a sifter (Bühler, Uzwil, Switzerland)
by using 100 g of semolina and a 5 min sifting time.
Moisture, ash and protein content in kernels were determined according to the official EU
methods of analysis [1]. Spaghetti were produced with all of the three different semolinas by
using the same operating conditions: recipe, mixing, extrusion process and low-temperature
drying process. The dried spaghetti were characterized for their diameter, hardness and colour.
The optimum cooking time (OCT) was determined by the white core disappearance [2].
Weight increase, diameter increase, cooking loss and total organic matter released in rinsing
water were determined at different times during cooking [2] and overcooking (OCT + 25%).

RESULTS & DISCUSSION

The semolinas with lower particle size showed higher ash, protein and gluten content, a higher
gluten extensibility but a lower gluten index and yellow colour than coarse semolina (Table 1).

Table 1. Chemical and physical properties of semolinas

Particle Ash Protein Gluten Gluten Gluten Colour
size (Pm) (g/100gdw) (g/100gdw) (g/100gdw) index extensibility (mm) b*
275 0.88a 14.17a 11.54 91 28 27.1
375 0.58c 13.48b 10.50 94 6 34.6
475 0.65b 13.25b 9.70 95 3 35.9

11th International Congress on Engineering and Food - Athens, Greece, 2011 2205
\
Figure 1. Hardness of spaghetti from different semolinas during cooking and overcooking time.

Spaghetti from coarse and medium-coarse semolina showed higher optimum cooking time (18
min) than spaghetti from medium semolina (15 min). Cooking time being equal, the weight
and diameter increase was higher in spaghetti from coarse semolina.
Within the optimum cooking time the hardness (maximum shear stress) of spaghetti from
coarse and medium-coarse semolina (Figure 1) were higher than those from medium semolina;
on the contrary the latter were harder than the former in overcooking, likely due to the higher
gluten content of medium semolina. When the shear stress was expressed as a function of the
normalized weight increase no differences were observed between the different samples.
No differences in cooking loss and total organic matter released after rinsing were found
among samples. The colour of cooked spaghetti from coarse semolinas were lighter and less
saturated than those from medium semolina due to the higher water uptake of the formers.

CONCLUSION
The high OCT, hardness and shear stress (before OCT) of the semi-cooked pasta obtained from
coarse and medium coarse semolina could be useful in two-step cooking processes in which
pasta is pre-cooked and cooled before the final cooking step. These are cooking processes
widely used in restaurants and catering industry.

REFERENCES
[1] EEC 2000. Commission Regulation 824/2000. Procedures for the taking-over of cereals by
intervention agencies and laying down methods of analysis for determining the quality of cereals.
Official Journal of the European Community, L100/3(20.4.2000), 31.
[2] Cocci E., Sacchetti G., Vallicelli M., Angioloni A. & Dalla Rosa, M. 2008. Spaghetti cooking by
microwave oven: cooking kinetics and product quality. Journal of Food Engineering, 85(4), 537-546.

2206
 Kinetics of heterogeneous amylolysis in oat flour and characterization of hydrolyzates

Anna Patsioura *a, Vassilis Gekas b, Athina Lazaridou c, Costas Biliaderis c

a
Department of Environmental Engineering, Technical University of Crete, Chania, Greece, email:
annapatsioura@yahoo.gr
b
Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of
Technology, Lemesos, Cyprus, e-mail: vassilis.gekas@cut.ac.cy
c
Department of Food Science and Technology, School of Agriculture, Aristotle University, Thessaloniki,
Greece, e-mail: athlazar@agro.auth.gr, biliader@agro.auth.gr

INTRODUCTION
Oat-based products with desirable sensorial properties contribute to the maintenance of human health;
their important nutritional attributes have been related to control of blood glucose and cholesterol levels.
As a nutrient source, oat grains offer a balance between carbohydrates, protein and fat, thus meeting
human nutritional needs very well; as a result, there are several types of newly developed oat-based
products with enhanced health properties on the market. For example, ‘oat milk’, a hydrolysis product of
oat flour, is offered as an alternative to soya or cow’s milk, particularly for people suffering from lactose
intolerance or for individuals aiming at a better control through their diet of the blood serum cholesterol
and the postprandial glucose and insulin levels.
In the case of oat flour aqueous dispersions, studies on their enzyme hydrolysis kinetics in conjunction
with the rheological characterization are of major interest for a thorough understanding of their
technological and physiological functionality. The objective of this work was to evaluate the enzyme
hydrolysis kinetics of oat flour aqueous dispersions under varying conditions and to characterize the
respective hydrolyzates.

MATERIALS & METHODS

Oat flour samples and two commercial enzyme preparations, Į-amylase and ȕ-amylase (OATLY AB,
Landskrona, Sweden), were employed for the starch hydrolysis studies. The hydrolysis kinetics of the oat
flour dispersions were examined at a solids concentration of 10% (w/w) under continuous mechanical
stirring at 60 oC (for 3 h). The oligosaccharides produced (glucose, maltose, maltotriose and
maltotetraose) were determined at various sampling intervals using an HPLC method, as described by
Hatzikamari et al. [1]. Steady shear rheological measurements at 200 s-1 were performed to monitor the
time dependence of viscosity changes during enzymic hydrolysis using a rotational Paar Physica MCR
300 rheometer.

RESULTS & DISCUSSION

The hydrolysis data indicated that maximum conversion of starch to maltose (~60 g maltose/ 100 g
starch) occurs when both Į- and ȕ-amylases are involved in the enzymic reaction mixture (Fig. 1); the
second major oligosaccharide produced was maltotriose (~ 10 % w/w starch basis). Instead, by the action
of Į-amylase alone, the main products were maltose (35% starch basis) and maltotriose (25% w/w starch
basis), and only minor amounts of glucose and maltotetraose were present in the hydrolyzates after the
first hour of hydrolysis (data not shown).

11th International Congress on Engineering and Food - Athens, Greece, 2011 2207
 (a) 70
60

g oligosaccharides/
100g of oat starch
50
40
30
20
10
0
0 20 40 60 80 100 120 140 160 180 200

Time (min)

Figure 1. Enzyme hydrolysis kinetics of oat flour aqueous dispersions (200 mL of 10 % w/w flour slurries) at 60 oC,
by the action of: Į-amylase (4.16 FAU) and ȕ-amylase (57oL) (Symbols - ‘:maltose; Ŷ:maltotriose).

Depending on the enzyme(s) type and concentration used, a reduction in viscosity was observed
throughout hydrolysis, with the Į-amylase alone exhibiting a greater thinning impact on the heated oat
flour slurries (Fig. 2). The viscosity profiles reflect a composite result of gelatinization (granule swelling)
– solubilization of the starch granules (viscosity rise, first 30 min) and thinning of the flour dispersion due
to enzyme hydrolysis of the solubilized starch components.

(a) 0.08
0.07
0.06
0.05
Ș (Pa s)

0.04
0.03
0.02 ȕ-amylase
0.01 Į-amylase : ȕ-amylase
0 Į-amylase
0 20 40 60 80 100 120 140 160 180 200
Time (min)

Figure 2. Comparison of the viscosity profiles during enxymic hydrolysis (60 oC, 200 s-1) of oat flour aqueous
dispersions (200 mL of 10 % w/w flour slurries) at 60 oC, with Į-amylase (8.3 FAU), ȕ-amylase (6oL) & both Į-
amylase (0.416 FAU) and ȕ-amylase (5.7oL).

CONCLUSION
The kinetic profiles of sugars presented above exhibit that maltose production mainly happens when the
two amylases are both involved in the enzymic reaction. The rheological measurements have shown an
extensive reduction in viscosity when the Į-amylase alone was applied. On the contrary, when ȕ-amylase
(exo-acting hydrolase) was used either by itself or in combination with Į-amylase the reduction in
viscosity was milder and reached a steady value after the 1st hour of hydrolysis.

REFERENCES
[1] Hatzikamari M., Kyriakidis D.A., Tzanetakis N., Biliaderis C.G. & Litopoulou-Tzanetaki E. 2007. Biochemical changes
during a submerged chickpea fermentation used as a leavening agent for bread production. European Food Research and
Technology, 224, 715-723.
[2] Nascimento J.R.O., Junior A.V., Bassinello P.Z., Cordenunsi B.R., Mainardi J.A., Purgatto E. & Lajolo F.M. 2006. Beta-
amylase expression and starch degradation during banana ripening. Postharvest Biology and Technology, 40, 41-47.

2208
 Kinetics of Amycolatopsis mediterranei DSM 43304 lipase-mediated synthesis of isoamyl

acetate in n-hexane

Dharmendra .S. Dheemana, Jesús M. Fríasb, Gary T.M. Henehana

a
School of Food Science & Environmental Health, Dublin Institute of Technology, Cathal Brugha Street,
Dublin 1, Ireland (dheeman@gmail.com, jesus.frias@dit.ie, gary.henehan@dit.ie )

INTRODUCTION
Esters of short-chain fatty acids are important flavour and fragrance compounds that are widely
used in the food and beverage industries. Isoamyl acetate is the character impact compound of
banana flavour and pear drops. It is one of the most highly employed compounds (74 tonnes
per annum) in the food industries [1]. Consequently, enzymatic synthesis of isoamyl acetate
and other aroma active esters is of increasing relevance to the food industry [2]. A number of
commercial lipases have been employed for direct esterification and transesterification in
organic solvents to produce isoamyl acetate [3]. However, few attempts have been made to
synthesize isoamyl acetate using non-commercial lipases [4]. The objective of the present
investigation was to test the performance of a Celite-immobilized A. mediterranei lipase
(AML) for the synthesis of isoamyl acetate in n-hexane and model its kinetics.

MATERIALS & METHODS

AML catalysed synthesis of isoamyl acetate was carried out in a batch stirred reactor with a
ase spherical geometry and 50 mL capacity. The reactor equipped with a magnetic stirrer,
containing 300 mM of isoamyl alcohol and 750 mg immobilized lipase as catalyst in n-hexane
were placed in a thermostatic water bath at 37± 0.1 °C and a stirrer speed of 200 rpm. Acetic
acid (300 mM) was then added to initiate the reaction. The concentration kinetics were fitted to
the kinetic models using the Levenberg-Marquardt nonlinear regression from ODRPACK. The
differential equations resulting from the different ester synthesis mechanism proposed were
simulated using the ODEPACK library [5].

RESULTS & DISCUSSION

A sequential strategy of experimental design proved to be useful in determining the conditions
for maximizing the equilibrium conversion in n-hexane using Celite-immobilized AML as a
catalyst. Optimum conversion was obtained at an acetic acid/isoamyl alcohol molar ratio of 2,
initial addition of 1.0% (v/v) of water and 7.5% (w/v) of enzyme (i.e. 2.5 g of enzyme mol-1 of
alcohol) at 50 °C. Under these conditions, a 12 h reaction time was sufficient to reach the
equilibrium molar conversion of 59%; however under non-optimized operational conditions the
equilibrium molar conversion reached was 21% after 36 h of reaction time.
Widely different conversion yields of isoamyl acetate in organic solvent systems have been
reported in the literature. A maximum conversion yield of 100% was reported by Romero et al.
[2] using C. antarctica Novozyme 435 at 13.8 g mol-1 of substrate in n-hexane, whereas
Liaquat and Owusu Apenten [4] used n-hexane during esterification to obtain a conversion
yield of <30% in 48 h with plant seedling lipases at 200 g mol-1 of substrate. In the present

11th International Congress on Engineering and Food - Athens, Greece, 2011 2209
investigation a non-commercial Celite-immobilized AML at 2.5 g mol-1 of substrate was
employed to achieve a conversion yield of 59% in 12 h, indicating a significant esterification at
a much lower enzyme concentration.
Esterifications of various organic acids with different alcohols by a variety of commercial
lipases are often modelled using the so called Ping Pong Bi Bi mechanism, a well known and
widely accepted mechanism for lipase-catalyzed reactions [3, 4]. Following the methodology
of Paiva et al. [4], the Michaelis-Menten dissociation constant terms for each of the compounds
from the enzyme complex were considered for model reduction.Each proposed models was
separately fitted to the experimental data and F-tests were performed with the aim of
investigating the statistical likelihood of such simplifications. The resulting rate expression is
described by Eq. (1).
­°
{( k cat f [ E t ]) u ( k cat r [ E t ])} u ®>$c @u >ǿǹǹ @ 
>ǿǹǹ c @u >+ 2 2 @½°
¾
°̄ k eq °¿
r (1)
( k cat f [ E t ])
( k cat r [ E t ]) >$c @u >ǿǹǹ @+ >ǿǹǹ c @u >+ 2 2 @
k eq

where r is rate constant (mol L-1 h-1), [Et] is total enzyme concentration (g L-1), [Ac], [IAA],
[IAAc] are acetic acid (M), isoamyl alcohol (M) and isoamyl acetate (M), respectively. kcatf and
kcatr are forward and reverse catalytic efficiency (h-1) of the AML and keq is the equilibrium
constant (dimensionless) of the reaction.

CONCLUSION
The present study is the first report delineating kinetics of direct esterification reaction between
a short-chain acid and isoamyl alcohol to synthesise isoamyl acetate using a non-commercial
Celite®545 immobilized lipase from an actinomycete strain. Optimized conditions for the
synthesis of isoamyl acetate were 7.5% (w/v) of Celite-immobilized AML, an acid/alcohol
molar-ratio of 2 with an initial addition of 1% (v/v) water at 50 °C and 200 rpm. Under these
conditions the equilibrium conversion yield obtained was 59% in 12 h. A simplified model,
based on a postulated Ping Pong Bi Bi mechanism, adequately described the kinetics of Celite-
immobilized AML catalysed direct esterification of isoamyl alcohol with acetic acid. Future
experiments, exploiting statistical process optimization designs, are likely to be able to raise
productivity further, making this lipase a potential candidate for the production of isoamyl
acetate.

REFERENCES
[1] Welsh F.W., Murray W.D., Williams R.E. 1989. Microbiological and enzymatic production of flavor
and fragrance chemicals. Critical Reviews in Biotechnology 19, 105–169.
[2] Romero M.D., Calvo L., Alba C., Daneshfar A. 2007. A kinetic study of isoamyl acetate synthesis by
immobilized lipase-catalyzed acetylation in n-hexane. Journal of Biotechnology, 127, 269–277.
[3] Liaquat M. & Owusu Apenten R.K. 2000. Synthesis of low molecular weight flavour esters using
plant seedling lipases in organic media. Journal of Food Science, 65, 295–299.
[4] Paiva A.L., Van Rossum D., Malcata F.X. 2002. Kinetics of lipase-mediated synthesis of butyl
butyrate in n-hexane. Biocatalysis & Biotransformation, 20, 43–51.
[5] Radhakridhnan K. & Hindmarsh A. 1993. Description and Use of LSODE, the Livermore Solver for
Ordinary Differential Equations. Lawrence Livermore National Laboratory report UCRL-ID-113855.

2210
 PROBIOLIVES: Table olive fermentation with selected strains of probiotic lactic acid
bacteria. Towards a new functional food (FP7-SME-2008- 2 project)

Chrysoula C. Tassoua, Efstathios Z. Panagoub, Antonio Garrido- Fernandezc, Cidalia Peresd, Luca
Cocoline & Nadia Chammemf

a
National Agricultural Research Foundation, Institute of Technology of Agricultural Products, Athens,
Greece (ctassou@nagref.gr)
b
Agricultural University of Athens, Dept. of Food Science & Technology, Lab. Of Microbiology &
Biotechnology of Foods, Athens, Greece (stathispanagou@aua.gr)
c
Instituto de la Grasa, Consejo Superior de Investigaciones Científica, Seville, Spain (garfer@cica.es)
d
Instituto Nacional dos Recursos Biológico, Lisbon, Portugal (cperes@itqb.unl.pt)
e
University of Turin, Faculty of Agriculture, Sector of Microbiology and Food Science, Turin, Italy
(lucasimone.cocolin@unito.it)
f
L'Institut National des Sciences Appliqées et de Technologie,Tunis, Tunisia (chnadia@yahoo.fr)

INTRODUCTION
The current project is a collaborative action of 14 participants. There are 4 SME-AGs
(PEMETE -Greece, ASEMESA - Spain, APABI - Portugal, AIFO - Italy) comprising of table
olive producing SMEs and 4 industries producing fermented olives from the main olive
producing countries (OLYMP-Greece, JOLCA-Spain, AZAGAP-Italy and PROBEIRA-
Portugal), 3 Research Institutions from Greece (NAGREF), Spain (IG-CSIC) and Portugal
(INRB); and 3 Universities from Greece (AUA), Italy (UNITO) and Tunisia (INSAT), all of
which are selected for their multidisciplinary individual expertise in the proposed research as
well as for their ability to provide expert facilities to perform the various tasks.
The concept of this project is to provide to the SME Associations and their members SMEs
with tools to increase their technological level, competitiveness and profits by the production
of olives, fermented with probiotic bacteria, preferably isolated among the lactic acid bacteria
colonizing the olives. Lactic acid bacteria bacteria from the olive microbiota are the dominant
microorganisms in natural fermentations and there will be studied if some of them possess
probiotic properties. The selected probiotic bacteria will be introduced into the brines at the
onset of fermentation, to act as starters [1,2], to be able to dominate and ensure a proper
fermentation inhibiting the growth and survival of undesirable microorganisms.
The goal is the production of a functional product, containing probiotic bacteria in adequate
amounts to improve consumer’s health, without altering the quality characteristics of fermented
olives. Consumer acceptance studies will be essential for the exploitation and the introduction
of the new food into the EU and international market. At the same time a better control of the
fermentation process, early detection of faulty fermentation and spoilage and assessment of the
time needed for fermentation completion will be achieved by monitoring the quality indices
[3] throughout the process with the use of advanced and emerging instruments and tools
(mathematical models) [4].

11th International Congress on Engineering and Food - Athens, Greece, 2011 2211
MATERIALS & METHODS
The work plan of the project includes the following: a) Characterization of the olive microbiota
and selection of probiotics. b) Use of the selected strains as starters in olive fermentations. c)
Evaluation of the shelf life of the final fermented product under different storage conditions. d)
Application of mathematical tools to predict the fermentation kinetics and survival of the
probiotic lactic acid bacteria. e) Safety of the probiotic fermented olives and risk analysis. f)
Consumer studies and g) application of the most suitable strains and processes in test field
studies.

RESULTS & DISCUSSION

The PROBIOLIVES project is in progress running its second year. The results so far are very
promising. A great number of lactic acid bacteria has been isolated from different cultivars in
Greece, Spain, Portugal, Italy and Tunisia. The tests that have been performed in vitro for their
probiotic potential have indicated that certain lactic acid bacteria have shown probiotic
properties. The most promising ones have been used as starters in olive fermentations in each
participating country. The fermentations have been monitored with microbiological and
physicochemical analyses performed at regular intervals and the data are under study.
Molecular techniques have also been used to detect the potential probiotic strains in the final
fermented product. Packaging and safety studies are also in progress.

CONCLUSION
There are very promising results regarding the isolation of new probiotic strains of lactic acid
bacteria from olives that are suitable to be used as starter cultures in olive fermentation.

REFERENCES
[1] Panagou, E.Z., Tassou, C.C. and Katsaboxakis, K.Z. 2003. Induced lactic acid fermentation of
untreated green olives of the Conservolea cultivar by Lactobacillus pentosus. Journal of the Science
of Food and Agriculture, 83,667-674.
[2] Pintado, C., Brito, D., Catulo, L., Peres, F., & Peres, C. 2008. Lactobacillus pentosus DSM 16366
starter added to brine as freeze-dried and as culture in the nutritive media for Spanish style green
olive production. Grasas y Aceites 59(3), 232-236.
[3] Arroyo López, F.N., Bautista Gallego, J., Chiesa, A., Durán Quintana, M.C., Garrido Fernández, A.
2009. Use of a D-optimal mixture design to estimate the effects of diverse chloride salts on the
growth parameters of Lactobacillus pentosus. Food Microbiology 26, 396-403.
[4] Panagou, E.Z., Tassou, C.C., Saravanos, E. and Nychas, G.-J.N. 2007. Application of neural
networks to simulate the growth profile of lactic acid bacteria in green olive fermentation. Journal of
Food Protection, 70, 1909-1916

2212
 Effect of vacuum drying on blackcurrant’s antioxidant components

Mónika Stéger-Máté, Beatrix Nótin, Réka Juhász, Balázs Verasztó, Dávid Jakab, Judit Monspart-Sényi,
József Barta

Corvinus University of Budapest, Faculty of Food Science, Department of Food Preservation, Budapest,
Hungary (monika.stegernemate@uni-corvinus.hu)

INTRODUCTION
Food components (vitamins, coloring agents, antioxidants, mineral components etc.) are
essential for healthy function of human organism, for prevention or medication of certain
diseases. Black currant fruit (Ribes nigrum L.) contains high amount of biologically active
components (vitamin C, anthocyanins, polyphenols, minerals, etc.) beneficial for human health.
Fresh consumption of black currant is not typical, it is mainly distributed as processed food
(concentrate, jam, frozen, dried powder etc.). For these reasons effect of preservation
technologies on changes of valuable components has primarily importance [1,2,3].
Aim of present study was to investigate changes of antioxidant compounds (vitamin C, total
polyphenol, total anthocyanin,) and antioxidant activity of black currant during vacuum drying
at different temperature levels (40-50-60oC). As for control atmospheric drying at 60°C was
also performed.

MATERIALS & METHODS

Black currant (Ribes nigrum L.) var. Titania grown in Hungary in 2010 was used for the
experiments. Samples were dried at three temperature levels (40-50-60 oC at 10 mbar) as long
as they reached a wet content lesser than 10%. Vacuum drying was performed by an industrial
Memmert V 200 vacuum dryer. As a control, atmospheric drying was performed at 60°C using
an atmospheric dryer. During drying process were determined the dry material content, total
polyphenol (TPC), total anthocyanin (TAC), ascorbic acid (AA) and antioxidant activity
(FRAP). Statistical evaluation was performed using Statistica 9. Software.

RESULTS & DISCUSSION

Initial wet content was 79.62 %. In case of atmospheric drying at 60°C took 16 hours to reach
final value, while using vacuum drying at same temperature level it was only 8 hours. When
vacuum drying was performed decreasing drying temperature (60-50-40°C) resulted in longer
drying time (8-10-12 hours).
Total polyphenol, total anthocyanin, and ascorbic acid content and antioxidant activity of black
currant expressed in mg g-1 dry material decreased compared to initial values during each
drying method.
Antioxidant compounds and antioxidant activity of black currant were affected by drying
temperature and pressure. Lowest FRAP values of end product were measured in case of 40°C
and 60°C vacuum dried samples (0.04 mg g-1 dry material and 0.08 mg g-1 dry material,
respectively), while at 50°C vacuum and atmospheric drying there was no significant
difference between initial and final FRAP values (Table 1).
50°C vacuum drying proved to be the optimal for preserve antioxidant compounds among the
dehydration methods investigated.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2213
Table 1. Antioxidant activity and compounds of black currant during different drying methods
Sample Time (h) FRAP TPC TAC AA
mg As g-1 DM mg g-1 DM mg g-1 DM mg g-1 DM
X±SD X±SD X±SD X±SD
Fresh 0 0.13±0.00 20.26±0.03 3.62±0.14 7.6±0.11
2 0.09±0.01 10.43±0.45 2.67±0.01 3.2±0.01
4 0.07±0.02 8.44±0.01 2.06±0.07 3.3±0.02
40 ºC 6 0.09±0.01 5.88±0.16 1.74±0.08 2.6±0.02
8 0.06±0.01 6.19±0.28 1.39±0.07 3.2±0.01
10 0.04±0.01 5.62±0.31 1.43±0.08 3.7±0.02
12 0.04±0.00 5.39±0.13 1.64±0.05 3.9±0.00
2 0.13±0.00 18.70±0.31 4.52±0.05 4.5±0.00
4 0.14±0.03 17.34±0.21 5.78±0.09 2.2±0.03
50 ºC 6 0.12±0.01 16.49±0.25 4.64±0.15 1.8±0.10
8 0.12±0.01 12.33±0.14 4.16±0.02 2.2±0.00
10 0.11±0.02 11.79±0.56. 3.69±0.31 2.6±0.06
2 0.09±0.01 16.78±0.78 3.78±0.02 4.9±0.01
4 0.10±0.03 13.42±0.16 3.99±0.69 2.3±0.03
60 ºC 6 0.08±0.00 15.79±0.09 3.44±0.02 2.9±0.06
8 0.08±0.00 15.58±0.07 3.72±0.16 2.6±0.04
2 0.10±0.00 15.79±0.20 4.65±0.13 6.6±0.20
4 0.12±0.01 15.68±0.76 4.29±0.07 4.5±0.46
6 0.12±0.00 15.79±0.31 3.49±0.14 5.2±0.05
60 ºC atm. 8 0.13±0.01 16.70±0.49 3.14±0.06 4.1±0.03
10 0.14±0.00 16.55±0.56 3.19±0.09 3.8±0.00
12 0.14±0.01 15.61±5.03 2.96±0.28 3.4±0.14
14 0.14±0.01 17.76±0.10 2.24±0.16 3.2±0.10
16 0.14±0.00 17.79±0.58 1.95±0.12 3.0±0.02

CONCLUSION
Based on our results it was concluded that drying temperature affects drying duration, rate of
wet decrease and amount of antioxidant compounds. Drying temperature at 50°C proved to be
the optimum. Black currant products made by moderate drying technology are suited for
production of functional food products (for example: mueslies, teamixes, sauces with fruit
pieces, jams etc.) due to their high antioxidant content.

The authors acknowledge the financial help of the TÁMOP 4.2.1./B-09/1/KMR-2010-0005 grant.

REFERENCES
[1] Souci S.W., Fachmann W. & Kraut H. 2008. Die Zusammensetzung der Lebensmittel Nährwert-
Tabellen 7. Revidierte Auflage, Medpharm Scienctific Publishers, Stuttgart, Deutschland, 1071.
[2] Slimestad R. & Solheim H. 2002. Anthocyanins From Black Currants (Ribes nigrum L.). Journal of
Agricultural and Food Chemistry, 50, 3228-3231.
[3] Mcdougall G.J., Gordon S. & Brennan R. 2005. Anthocyanin-Flavanol Condensation Products From
Black Currant (Ribes nigrum L.). Journal of Agricultural and Food Chemistry, 53, 7878-7885.

2214
 Production of Bioactive Metabolites with Pharmaceutical and Nutraceutical Interest by

Submerged Fermentation of Pleurotus ostreatus in a Batch Stirred Tank Bioreactor

Lefki-Maria Papaspyridia, Nektarios Aligiannisb, Paul Christakopoulosa, Alexios-Leandros Skaltsounisb

Nikolas Fokialakisb,

a
BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical
University of Athens, 9 Iroon Polytechniou Street, Zografou Campus, 15700 Athens, Greece,
leriapap@mail.ntua.gr
b
Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of
Athens, Panepistimioupolis, 15771, Athens, Greece,
fokialakis@pharm.uoa.gr

INTRODUCTION
Mushrooms comprise a vast and yet largely untapped source of powerful new pharmaceutical
products having potential as functional foods and sources of novel molecules [1]. Pleurotus
ostreatus (Jacq.:Fr.) P. Kumm., also known as the oyster mushroom is a basidiomycete
belonging to the family Pleurotaceae (Agaricales, Agaricomycetes). In the last decade, the
scientific interest in this species has increased considerably because of its gastronomic value
and its nutraceutical and pharmaceutical properties [2]. Natural products from mushrooms are
mostly obtained through the field-cultivation of the fruiting bodies. However, the submerged
fermentation of their mycelial form has received much attention as a promising alternative for
efficient production of their biomass and active metabolites [3]. The aim of this study was the
isolation and identification of bioactive metabolites derived from biomass produced by
submerged fermentation of the edible mushroom P. ostreatus (strain ATHUM 4438) in a batch
stirred tank bioreactor.

MATERIALS & METHODS

The composition of culture medium and fermentation conditions of P. ostreatus ATHUM 4438
(obtained from the ATHUM Culture Collection of Fungi of University of Athens) in bioreactor
used, was the suggested for maximum biomass production, reported in our previous study [4].
The mycelia biomass was extracted using the method of Accelerated Solvent Extraction (ASE).
The crude dichloromethane extract (DCM) was initially fractioned by means of medium
pressure liquid chromatography (MPLC). In order to extract efficiently the phenolic
compounds of the initial methanolic extract (MeOH), an adsorption-desorption process using
XAD-4 type resin as efficient sorbent, was performed. The isolation procedure of the resulting
phenolic fraction was carried out by Fast Centrifugal Partition Chromatography (FCPC).
Further analysis of all the resulting fractions was conducted by means of Sephadex LH-20
column chromatography, preparative HPLC and TLC. All isolates were identified by 1D/2D
NMR-spectroscopic analyses, NMR data comparisons, comparison with literature data and
chemical correlations combined with GC/MS and LC/MS experiments.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2215
RESULTS & DISCUSSION
The main metabolite chemotypes present in extracts of mycelia biomass produced by
submerged fermentation process of P. ostreatus ATHUM 4438 in a stirred tank bioreactor were
fatty acids, phenolic metabolites, nucleotides, and alkaloids. Specifically, the compounds
afforded by the DCM extract were identified as linoleic acid (1), oleic acid (2), stearic acid (3),
palmitic acid (4) and their corresponding methyl esters (5, 6, 7 and 8, respectively), benzoic
acid (9), trans 3, 4-dihydro-3, 4, 8-trihydroxynapthalen-1(2H)-one (10), 4-hydroxy-
benzaldehyde (11), indolo-3-carboxylic acid (12) and uracil (13).

Considering the phenolic compounds in the MeOH extract, it was observed that almost 14% of
it consists of such compounds. The investigation of the phenolic extract (POXM), based on the
effective fractionation by FCPC analysis, afforded 3-formyl-pyrrole (14), 4-hydroxy-benzoic
acid (15), uridine (16), nicotinic acid (17) and nicotinamide (18).

Based on existing literature data, compounds (1-8), (17) and (18) are regarded as functional
food ingredients, exerting numerous health benefits (e.g. lowering heart attack risk and
arteriosclerosis and aiding in cancer prevention), while the other metabolites obtained in this
study are considered of great pharmaceutical interest, demonstrating great biological activities
(e.g. exhibiting anti-inflammatory and antioxidant properties). An interesting observation was
that compounds (10), (12), (14) and (15) have not been reported again from this mushroom
strain, revealing its potential for the development of powerful new pharmaceutical or
nutraceutical products.

CONCLUSION
From the data presented herein, it is proved that P. ostreatus ATHUM 4438 may be used as
potential source of bioactive metabolites for food supplements or in the development of
pharmaceuticals. Most importantly, it is demonstrated that the established fermentation process
of the studied P. ostreatus strain in a batch stirred tank bioreactor is viewed promising for this
objective to be carried out on industrial scale.

REFERENCES
[1] Ferreira I.C.F.R., Vaz J.A., Vasconcelos M.H. & Martins A. 2010. Compounds from wild edible
Mushrooms with Antitumor Potential. Anti-cancer Agents in Medicinal Chemistry, 10(5), 424-436.
[2] Gregori A., Švagelj M. & Pohleven J. 2007. Cultivation Techniques and Medicinal Properties of
Pleurotus spp. Food Technology and Biotechnology, 45(3), 238-249.
[3] Tang Y.Z., Zhu, L.W., Li H.M. & Li D.S. 2007. Submerged culture of mushrooms in bioreactors-
challenges, current-state-of-the-art, and future. Food Technology and Biotechnology, 45(3), 221-229.
[4] Papaspyridi L.-M., Katapodis P, Gonou-Zagou Z., Kapsanaki-Gotsi E. & Christakopoulos P. (2010).
Optimization of biomass production with enhanced glucan and dietary fibres content by Pleurotus
ostreatus ATHUM 4438 under submerged culture. Biochemical Engineering Journal, 50(3), 131-138.

2216
 Effects of High Intensity Pulsed Electric Fields or Thermal Treatments on Carotenoid

Profile of a Fruit Juice-Soymilk Beverage along Chilled Storage

Laura Salvia-Trujillo, Mariana Morales-de la Peña, Ma. Alejandra Rojas-Graü, Olga Martín-Belloso

University of Lleida, Lleida, Spain (lsalvia@tecal.udl.cat)

INTRODUCTION
Carotenoids are considered potential antioxidants and free radical scavengers which modulate
the pathogenesis of cancers [1] and coronary heart diseases [2]. Daily consumption of products
rich on these compounds, such as fruits and vegetables, is highly recommended. Nowadays,
mixed beverages containing fruit juices and soymilk are receiving considerable attention due to
their high content of functional ingredients such as vitamin C and isoflavones; having, at the
same time, high antioxidant capacity [3, 4]. Moreover, being a blend of different fruit juices
and soymilk, they could contain considerable amounts of carotenoids.
Thermal pasteurization is known to effectively inactivate microorganisms and deleterious
enzymes of fruit beverages. Nevertheless, the high temperature achieved during processing
destroys most desirable health-compounds. Hence, high intensity pulsed electric fields (HIPEF)
have being under continuous investigation as a potential food preservation technology. It has
been demonstrated that microbial and enzymatic inactivation levels achieved by HIPEF can be
as high as those reached by heat and, in addition, HIPEF processing leads to better retention of
bioactive compounds in fruit and vegetable juices [3, 4, 5]. Unfortunately, there is no current
information related to carotenoid composition of FJ-SM beverages and how HIPEF or thermal
process affects it. The aim of this study, therefore, was to identify the carotenoid composition
of a fruit juice-soymilk (FJ-SM) beverage and evaluate the effects of HIPEF (35 kV/cm, 4 ȝs-
bipolar pulses at 200 Hz during 800 or 1400 ȝs) or thermal (90 ºC, 60 s) treatments over these
compounds during the storage at 4ºC.

MATERIALS & METHODS

The FJ-SM beverage was prepared according to Morales-de la Peña et al. (4). Carotenoids were
extracted and quantified by HPLC, following a procedure validated by Cortés et al. (5).

RESULTS & DISCUSSION

Carotenoids identified in fresh and treated FJ-SM beverages were cis-violaxanthin +
anteraxanthin, cis-anteraxanthin, anteraxanthin, lutein, zeaxanthin, Į- and ȕ-cryptoxanthin, and
Į- and ȕ-carotene. Among them, lutein (0.066-0.108 mg/100mL), zeaxanthin (0.056-0.076
mg/100mL) and E-carotene (0.012-0.021 mg/100mL) were present in the highest
concentration. Just after processing, a significant decrease of lutein (16-39%), zeaxanthin
(26%) and E-cryptoxanthin (23-43%) content was observed, while E-carotene concentration
rose (6-17%) only in HIPEF beverages. Hence, total carotenoid concentration, obtained by the
sum of individual carotenoid content, was significantly diminished after processing (Fig 1).
Throughout storage, anteraxanthin, lutein, zeaxanthin and E-carotene content of untreated and
treated beverages tended to decrease so that total carotenoid concentration diminished with

11th International Congress on Engineering and Food - Athens, Greece, 2011 2217
time (Fig 1). Even though there were no significant differences between samples, HIPEF
beverages for 800 or 1400 Ps showed slightly higher carotenoid concentration than that heat
processed. The degradation of carotenoid content in the FJ-SM beverage could be due to
oxidation or enzymatic reactions occurred during processing and storage.

0.4
TCC (mg /100 mL) 0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0 10 20 30 40 50 60
Storage time (days)

Figure 1. Total carotenoid compounds (TCC) of untreated (Ƈ), high intensity pulsed electric field (35
kV/cm with 4 ȝs bipolar pulses at 200 Hz for 800 („) or 1400 ȝs (ʆ)) and thermal (ŏ) (90 ºC, 60 s)
treated fruit juice-soymilk beverages throughout storage at 4ºC.

CONCLUSION
Nine carotenoids were identified in the FJ-SM beverage, being lutein, zeaxanthin and ȕ-
carotene those present in higher concentrations. HIPEF and heat treatments caused a significant
decrease of initial carotenoid concentration of the FJ-SM beverage. Overtime, it tended to
decrease regardless of the treatment applied. However, HIPEF treated beverages always had
higher carotenoid concentration than the heat treated ones. The application of HIPEF could be
a good alternative to obtain FJ-SM beverages with similar carotenoid profile than fresh
beverages.

REFERENCES
[1] Giovanucci E. 1999. Tomatoes, tomato-based products, lycopene and cancer: a review of the
epidemiological literature. Journal of the National Cancer Institute, 91, 317-331.
[2] Kritechevsky S.B. 1999. ȕ-carotene, carotenoids and the prevention of coronary hearth disease.
Journal of Nutrition, 129, 5-8.
[3] Morales-de la Peña M., Salvia-Trujillo L., Rojas-Graü M.A. & Martín-Belloso O. 2010. Impact of
high intensity pulsed electric field son antioxidant properties and quality parameters of a fruit juice-
soymilk beverage in chilled storage. LWT – Food Science and Technology, 43 (6), 872-881.
[4] Morales-de la Peña M., Salvia-Trujillo L., Rojas-Graü M.A. & Martín-Belloso O. 2010. Isoflavone
profile of a high intensity pulsed electric field or thermally treated fruit juice-soymilk beverage
stored under refrigeration. Innovative Food Science and Emerging Technologies, 11 (4), 604-610.
[5] Cortés C., Esteve M.J., Frígola A. & Torregrosa F. 2004. Identification and quantification of
carotenoid incluiding geometrical isomers and vegetable juices by liquid chromatography with
ultraviolet-diode array detection. Journal of Agricultural and Food Chemistry, 52, 2203-2212.

2218
 Amino Acid Composition of a Fruit Juice-Soymilk Beverage as Affected by High

Intensity Pulsed Electric Fields or Thermal Treatments during Storage

Mariana Morales-de la Peñaa, Laura Salvia-Trujillo a, Teresa Garde-Cerdán b, Ma. Alejandra Rojas-Graüa,
Olga Martín-Bellosoa

a
University of Lleida, Lleida, Spain (mmorales@tecal.udl.cat)
b
Public University of Castilla-La Mancha, Albacete, Spain

INTRODUCTION
Nowadays, fruit juice consumption has been increased mainly because their nutritional
properties and bioactive principles (vitamins, minerals, phenolic compounds, carotenoids and
amino acids). Additionally, soymilk has been appreciated as a good source of amino acids (AA)
necessaries for optimal wellness (1). AA not only have a nutritive value, but also provide
several health benefits such as antimutagenicity and reduction of blood sugar and coronary
hearth diseases (2). At present, development of mixed beverages containing fruit juices and
soymilk has been attempted to overcome the typical beany-flavor of soy-based products.
Commonly, this type of beverages is preserved by heat pasteurization. However, the high
temperature achieved in the process generally causes undesirable health-attributes losses.
Hence, in order to avoid the detrimental effects caused by heat, non-thermal technologies, such
as high intensity pulsed electric fields (HIPEF), are becoming potential alternatives for liquid
food preservation. It has been reported that HIPEF process has minimal impact on nutritional
and sensory properties of foods and can extend their shelf-life by inhibiting microorganisms
and enzymes (3). Nevertheless, up to now there is no available literature regarding AA
composition of FJ-SM beverages and the effects of HIPEF or thermal process could have over
them. Hence, this work attempts to evaluate and compare the AA profile of a fruit juice-
soymilk (FJ-SM) immediately after HIPEF (35 kV/cm, 4 ȝs-bipolar pulses at 200 Hz during
800 or 1400 ȝs) or thermal (90 ºC, 60 s) treatments and during storage at 4ºC.

MATERIALS & METHODS

The FJ-SM beverage was prepared according to Morales-de la Peña et al. (4). AA determination
was carried out following the procedure described by Garde-Cerdán et al. (5).

RESULTS & DISCUSSION

Regardless of the treatment applied, aspartic acid, glutamic acid, serine, histidine, glycine,
threonine, arginine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine,
phenylalanine and lysine represented the initial free AA profile of the FJ-SM beverage.
Arginine (22 - 24 %) and proline (17 - 19 %) were the most abundant AA present in the
untreated and treated beverages, while isoleucine (0.88 – 1.1%) was considered the limiting
AA. Just after processing, HIPEF-800 Ps did not alter the individual AA content of the
beverage, although valine concentration increased. Otherwise, HIPEF-1400 Ps and thermal
treatments significantly affected the concentration of various AA. Thus, HIPEF-800 Ps
beverage showed a similar total AA content than the untreated one (64.5 - 64.79 mg/100mL),

11th International Congress on Engineering and Food - Athens, Greece, 2011 2219
whereas HIPEF-1400 Ps and heat processes diminished it up to 61.82 and 60.25 mg/100mL,
respectively. Overall, changes on the individual AA content of untreated and treated beverages
overtime led to an augment of the total AA concentration as storage time increased (Fig. 1).
Proteolysis reactions in the untreated and treated beverages might have occurred during the
storage allowing the increase on the concentration of most AA.
140

TFAA (mg/100 mL) 120

100

80

60

40

20

0
0 10 20 30 40 50 60
Storage time (days)

Figure 1. Total free amino acid (TFAA) content of untreated (Ƈ), high intensity pulsed electric field (35
kV/cm with 4 ȝs bipolar pulses at 200 Hz for 800 („) or 1400 ȝs (ʆ)) and thermal (ŏ) (90 ºC, 60 s)
treated fruit juice-soymilk beverages throughout storage at 4ºC.

CONCLUSION
Among the sixteen AA identified in the FJ-SM beverage, arginine and proline were present in
higher concentrations. Individual AA content was not affected by HIPEF (800 Ps) however,
immediately after HIPEF (1400 Ps) or heat treatments, the concentration of some amino acids
decreased. Along storage, most of the AA tended to increase, irrespectively of the treatment
applied, allowing that total AA content of the FJ-SM beverages gradually augmented with time.
Nonetheless, at the end of the storage HIPEF beverages had higher concentrations of AA than
that thermally processed. Hence, HIPEF is a potential process to obtain FJ-SM beverages with
stable shelf-life and similar AA composition than fresh samples.

REFERENCES
[1] Yang H. & Zhang L. 2009. Changes in some components of soymilk during fermentation with the
basidiomycete Ganoderma lucidum. Food Chemistry, 112, 1-5.
[2] Dajanta K., Apichartsrangkoon A., Chukeatirote E. & Frazier R. 2011. Free-amino acid of thua nao,
a Thai fermented soybean. Food Chemistry, 125, 342-347.
[3] Toepfl S., Heinz V. & Knorr D. 2005. Overview of pulsed electric field processing for food. In: Da-
Wen Sun (ed) Emerging Technologies for Food Processing.
[4] Morales-de la Peña M., Salvia-Trujillo L., Rojas-Graü M.A. & Martín-Belloso, O. 2010. Impact of
high intensity pulsed electric field son antioxidant properties and quality parameters of a fruit juice-
soymilk beverage in chilled storage. LWT – Food Science and Technology, 43 (6), 872-881.
[5] Garde-Cerdán T., Lorenzo C., Lara J.F., Pardo F., Ancín-Azpilicueta C., & Salinas M. R. (2009).
Study of the evolution of nitrogen compounds during grape ripening. Application to differentiate
grape varieties and cultivated systems. Journal of Agricultural and Food Chemistry, 57, 2410-2419.

2220
 Challenges and essentials for reinventing R&D in an open innovation ecosystem

I. Sam Saguy

The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of
Jerusalem, Rehovot, Israel (ssaguy@agri.huji.ac.il)

INTRODUCTION
Innovation is a company's lifeline to meeting the challenges imposed by global economic pressure,
unstable financial markets, and exponentially accelerating growth of scientific knowledge and
technological complexity, as well as consumers' needs and expectations. Innovation has a wide
spectrum of definitions and a multidimensional nature, touching upon every aspect of modern life.
Here, innovation is defined as the application of ideas, technology and processes in new ways to
gain a competitive advantage and create value [1]. Open innovation (OI) is a relatively new form of
interaction that that made significant inroads opening organizations to acquire, integrate and process
knowledge and to collaborate on co-development with external sources [2]. It offers acceleration by
reducing the burden of time pressure, while sharing human and physical resources and gaining a
critical mass of talented and highly skilled experts. It also benefits from embracing cultural
openness, networks, social impact and sharing the development risk [1,3]. This new mindset allows
knowledge to diffuse across company borders. However, despite OI's widespread application and
impact among large companies, small and medium-size enterprises (SMEs), particularly in the food
sector, are still struggling with its full implementation. The possibility that only 10% of all
companies have adapted themselves to OI [4] is alarming and highlights the significant challenges
we face in knocking down real and perceived roadblocks for reinventing R&D and reinnovating the
innovation process itself. The main objectives of this paper are to: underline some relevant OI
barriers and challenges, delineate essential changes and elements that should be adopted by both
industry and academia, and advocate a new approach to creating a "pull" force and platform for the
promotion and enhancement of collaborations and partnerships. Specific recommendations are
provided, with special emphasis on academia-industry's role and enhanced social responsibility.

DISCUSSION
Europe's competitiveness and its capacity to create new jobs, promote future growth and improve
standards of living depend on its ability to drive innovation in products, services, business and
social processes and models. This places innovation at the heart of the Europe 2020 strategy. It is
not surprising that the European Commission has identified innovation as the best means of
successfully tackling major societal challenges such as climate change, energy and resource
scarcity, health care and ageing. The untapped potential and full adaptation of the recently
burgeoning OI are relevant for the EU Food and Drink sector (F&D), especially for its SMEs.
Recent EU F&D 2008 data show that although SMEs comprise 99.1% of 308,000 companies and
62.8% of the employment, they generate only 48.2% of the turnover
(http://smes.ciaa.eu/php/index.php?doc_id=2). R&D investment represents 0.37% of F&D output,
lower than in most developed countries. Expanding the spectrum of companies that are able to
benefit from OI is a therefore significant challenge that calls for immediate action, involving several
paradigm shifts. One such paradigm is the pivotal role that academia should play in driving and co-

11th International Congress on Engineering and Food - Athens, Greece, 2011 2221
pursuing innovation, enhancing collaboration with industry and adding a new metrics of social
responsibility. Sharing-is-Winning principles [3] furnish industry and academia with an opportunity
to seek improved means and tools for the development of platforms that will maximize mutual
efforts leading to the creation of an innovation ecosystem. Academia needs to realize that
fundamental research is no longer sustainable as sole driving force. Applied research and becoming
"organic" members of the industrial efforts and teams are needed. Enhanced academia-industry
innovation interaction also calls for new approaches toward creating a bridge over the "Valley of
Death" (VoD) that exists between basic research and commercialization [5]. Academia management
must develop a strategy for sustaining a culture of promoting collaborations and enhancing the
status of applied R&D. Assessing the overall impact of research and inventions through the lens of
social contribution, and developing adequate metrics to quantify it are also recommended. Industry
also has a major role in this transformation. As most OI tools, know-how and practices have been
improved in the last decade, they are no longer an impediment. Management mindset, on the other
hand, can be, and as such it plays a vital role. In addition, to bring the process to fruition, industry
needs to clearly identify its needs at the outset, as well as the reasons why OI is the appropriate
model. Some points to consider include: benefits, cost, business model, people and expertise,
intellectual property (IP), time to market, and legal ramifications. SMEs should also consider the
unique obstacles that allow only partial utilization or complete blockage of OI: these can be related
to technology, product spectrum, ability to interact with large companies, adequate infrastructure,
available manpower, educational backgrounds, business functions, IP, etc. It is important to note
that although large industry is adept at implementing OI with or without academia, to make a
significant impact and/or improve SMEs' chances to be substantial OI players, academia needs to
serve as a catalyst.

CONCLUSION
Reinventing R&D in an OI ecosystem and increasing success rates in a growing competitive
marketplace require implementing significant new steps. The need for co-innovation with
complementary partners, alliances, enhanced collaboration, and the pivotal role of academia in
removing roadblocks for reinventing R&D are highlighted. These steps should reinnovate the
innovation process itself. OI is part of the innovation ecosystem and it is here to stay. Academia
should play a proactive role as innovation catalyst, bridging over real and perceived stumbling
blocks and establishing competence matching for co-development of innovation. Reinventing R&D
requires passionate people, committed executives and organizations, an innovation culture and
mindset, and communication. Mutual academic and industrial lenses will be utilized to draw
specific paradigm shifts, steps, and hands-on recommendations.

REFERENCES
[1] Traitler H., Watzke H.J. & Saguy I.S. 2011. Reinventing R&D in an Open Innovation Ecosystem.
Journal of Food Science, 76(2), R62-R68. [2] Chesbrough, H.W. 2003. Open Innovation: The New
Imperative for Creating and Profiting from Technology. Harvard Business School Press, Boston, MA,
USA. [3] Traitler H. & Saguy I.S. 2009. Creating Successful Innovation Partnerships. Food Technology,
63(3), 22-35. [4] Lindergaard S. 2010. The Open Innovation Revolution: Essentials, Roadblocks and
Leadership Skills. John Wiley & Sons, Hoboken, NJ, USA. [5] Markham S.K., Ward S.J., Aiman-Smith
L. & Kingon A.I. 2010. The Valley of Death as Context for Role Theory in Product Innovation. Journal
of Product Innovation Management, 27(3), 402-417.

2222
 Philosophy of an open R&D system

Kamel Chida

Generall Mills

Creating value for SMEs in the food industry through open innovation - Examples from

Norway

Øyvind Fylling-Jensen

Nofima AS

Open Innovation at Mars: Join-up, speed-up, scale-up

Olivier Fleurot

Mars GmbH

11th International Congress on Engineering and Food - Athens, Greece, 2011 2223
 Innovation sharing by cooperative R&D

Dieter Albers
Frutarom Savory Solutions GmbH, Korntal-Münchingen, Germany
(dalbers@frutarom.com)

The change in eating habits and consumers’ increasing expectations in terms of food quality
and safety also makes Frutarom Savory Solutions face ever new challenges if it wants to
continue to assert the significant market position it has obtained.
Frutarom Savory Solutions GmbH, as a Frutarom Group business unit, develops customised
solutions for the meat-,fish- and convenience food industry. The product portfolio comprises
spicy aromas, spice blends and starter cultures. Frutarom pursues acquisitions on a global basis
in order to further expand its technological know-how, product portfolio and its business
divisions‘ reach. It is Frutarom Savory Solutions GmbH’s express objective to become a
globally leading manufacturer in the market for spicy aromas and ingredients for meat-, fish-,
snack- and convenience food products.
Competitive products are the result of a careful and detailed development. The better the
analytic method, technology, process, and quality and safety management, the more successful
the company is with its customers. Therefore, Frutarom has recently decided to cooperate with
a strategic research partner, which is a non-profit research organisation, who is able to support
the company on an exclusive basis right from the beginning in all research and development
areas – be it the latest market trends or our customers’ special requests.
Driving force for this decision was not to call into question the companies own R&D
department but rather the question by how can Frutarom incorporate faster and more efficient
external knowledge in companies’ own products? How to establish a strategic cooperation with
external scientist with whom problems and needs have to be communicated without endanger
companies’ valuable own knowledge?
Therefor a prerequisite for the R&D cooperation was trust in the ability, expertise and
reliability of the external research and development provider. The cooperation with a non-
profit research organisation seems excellent as both parties collaborated before on project
level. The staff of the cooperating research institute also thinks outside their own box and use
diverse resources of knowhow and state-of-the-art equipment to finally be able to present
innovative solutions.
What is so special about this research collaboration is that the companies feel really connected.
The combination of the Frutarom Savory Solutions’ experts’ knowledge and the R&D provider
makes for practically relevant, directly applicable scientific research. Jobs at hand are
processed without any delay and the results are immediately fed into the product development
process. The time expended on new developments is much reduced: there is a much faster and
straighter response to market trends.
Steps towards this cooperation, expectations and first experiences will be presented showing
the benefit of knowledge and innovation sharing for both, the company as well as the external
research provider.

2224
 HighTech Europe Interactive Technology Portal – new tool
for innovation in food processing
Deutsches Institut für Lebensmitteltechnik e.V., Quakenbrueck, Germany (k.lienemann@dil-ev.de,
n.ay@dil-ev.de) (Lienemann Kerstin and Ay Nevaf), Wageningen UR - Food & Biobased Research,
Wageningen, The Netherlands (roos.groeneveld@wur.nl, don.willems@wur.nl) (Groeneveld Roos and
Willems Don), Katholieke Universiteit, Laboratory of Food Technology, Leuven, Belgium
(Iesel.VanderPlancken@biw.kuleuven.be) (Van der Plancken Iesel)

INTRODUCTION
22 partners from academia and industry are collaborating in the Network of Excellence HighTech
Europe (HTE, www.hightecheurope.eu) to develop tools for the stimulation of R&D and to boost
innovation in the food processing sector. The overall aim of the network is the establishment of a
European Institute for Food Processing (EU-IFP). One of the important building blocks of the EU-
IFP is the Interactive Technology Portal (ITP). The online portal will provide access to answers to
inquiries from industry, and demonstrate and communicate potential innovations in food processing.
The build-up of the portal makes use of the Science Cube, which is an approach to describe relations
between innovation sources, food processing operations, and underlying scientific principles. The
ITP is one of the novel approaches of HighTech Europe fostering open innovation in food
processing. Open innovation in this context means identifying and using opportunities for
innovation beyond the competences and resources available in ones’ own company or institute. The
objective of the HighTech Europe ITP is to transform complexity into simplicity and to provide a
portal that can and will be maintained after the end of the project.

MATERIALS & METHODS

An online portal is in general a single entry point for all kinds of information; a tool for bringing
people together and for providing information and solutions. The requirements for the HighTech
Europe ITP necessitated two main activities:
x Technical implementation of the ITP: Which semantic software provides the best solution for a
user-friendly knowledge portal? (A)
x Description and implementation of the screening procedure: How to identify the knowledge
input to the portal; how to illustrate and implement it; and how to describe a clear procedure
following the Science Cube approach? (B)
A) The ITP consists of three different building blocks: i) The SemanticMediaWiki software
inclusive SemanticForms [1], which is the backbone of the ITP. It is a free and very flexible open-
ended software application, written in the programming language PHP. ii) An ontology, which is a
formal representation of knowledge in the form of concepts from the projects target domain [2]. It
defines, furthermore, relationships between these concepts. An ontology can be used as a thesaurus,
it may be used to support information retrieval or information extraction, or even for automatic
reasoning. iii) The thesaurus-based full text search is an application used within the ITP
MediaWiki that uses the ontology as a thesaurus to support the user while searching through both
ITP and external documents. When a user of the ITP enters a term in the search box of the media
wiki, appropriate terms from the ontology are suggested in a drop down box and a full text search is
then performed on indexed documents. In addition, search terms are expanded using, broader and
narrower terms from the thesaurus/ontology.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2225
B) The HTE consortium considers in its screening procedure conventional and innovative food
processing technologies. On the one hand, the knowledge identified is used to further build up the
ontology, on the other hand, it is used to create technology, profile and infrastructure datasheets,
which are based on comprehensive literature searches (focus: review papers) and tacit knowledge
available at the beneficiaries’ institutions.
The Science Cube approach is used to characterize the knowledge in three different ways: i) by the
scientific principle it is based on (physical, chemical and biological), ii) by the food processing
operation it can be applied to (separation, stabilizing, structure forming and conversion processes
and packaging), and iii) by the innovation sources it can be attributed to (nanotechnology,
biotechnology, information and communication technology).

RESULTS & DISCUSSION

The ITP has been implemented (www.hightecheurope-portal.eu) and is now continuously updated.
Templates of datasheets have been created that meet the needs of the future user. Datasheets have
28 different entry fields (e.g. ’working principle’ or ‘risk or hazards’) whereas each entry field
presents a ‘property’ that facilitates a targeted search by the semantic search application of the ITP.
At the moment, technology datasheets (64), profile datasheets (205, not all complete), event and
infrastructure datasheets (being built up currently) are available. The ontology consists already of
more than 2,000 terms, which are grouped in the main concepts like “methods”, “products” or
“product characteristic”. The full text search function is implemented and gives the user suggestions
for broader, narrower or related terms. The user-friendliness and quality of the search function will
be increased by also offering a more detailed search using search and browse functions of the
MediaWiki software.

CONCLUSION
The MediaWiki based HighTech Europe Interactive Technology Portal is a promising portal for the
transfer and distribution of knowledge and technologies in the food processing area. During the
current implementation phase it is updated in content, quality, search functionality and user-
friendliness. Although access to the portal is restricted at the moment to project beneficiaries, it will
be opened to externals (members of the Associated Membership Platform) to test it, for further
input, and also to suggest improvements. The ITP will become an important tool for the European
food processing area to facilitate innovation. A next step in the network will be an open discussion
with stakeholders from industry, academia and policy how to maintain and improve it beyond the
end of the project.

ACKNOWLEDGEMENT: We wish especially acknowledge our colleagues of the HighTech Europe

consortium from CENTIV (DE), INRA (FR), IRTA (ES), SIK (SE), TTZ (DE), UTCN (RO) and VÚPP
(CZ). Furthermore we acknowledge Prof. Dr. Andreas Schmidt from the University of Applied Science in
Osnabrueck (DE) for providing us his expertise and experience in IT management.

REFERENCES
[1] http://semantic-mediawiki.org/wiki/Semantic_MediaWiki and
http://www.mediawiki.org/wiki/Extension:Semantic_Forms
[2] Gruber T. R. 1993, A translation approach to portable ontology specifications. Knowledge
Acquisition 5(2), 199–220.

2226
 Food Microstructure: a 3-D experience

B. Nicolai

Possibilities of X-ray nano-CT for internal quality assessment of food products

E. Herremansa *, S. Chassagne-Bercesb, H. Chanvrierb, A. Atoniukc, R. Kusztalc, E. Bongaersd, B.E.
Verlindene, E. Jakubczykf, P. Estradeg, P. Verbovena, B. Nicolaïa,e
a
Katholieke Universiteit Leuven, Willem de Croylaan 42, 3001 Leuven, Belgium
b
NESTEC SA, Nestle PTC. Route de Chavornay 3, 1350 Orbe, Switzerland
c
CHABER ltd, Prymasa Tysiclecia 83, 01242 Warsaw, Poland
d
SkyScan NV, Kartuizersweg 3b, 2550 Kontich, Belgium
e
VCBT, Flanders Centre of Postharvest Technology, Willem de Croylaan 42, 3001 Leuven, Belgium
f
SGGW, Warsaw University of Life Sciences, Dep. Food Eng. & Process MGMT, 02776 Warsaw, Poland
g
VSG, Visualization Sciences Group SAS, Avenue Kennedy 87, Mérignac Cedex 33708, France
*
Corresponding author electronic mail: els.herremans@biw.kuleuven.be

INTRODUCTION
Knowledge of food microstructure and how it changes during processing operations is essential
to produce high quality food. X-ray CT (Computed Tomography) uses X-rays to look inside
materials and produces 3D images. While micro-CT (X-ray imaging at micrometer resolution)
has become feasible over the last decade, many foods contain structural features (such as air
spaces, cells, cell walls) that are manifested over a large range of dimensions, including the
nanometer range. Up to date it has been nearly impossible to visualize structures on the
nanoscale in 3D with X-ray CT. As a consequence, it has been difficult to quantify the effects
of these nanoscale features on important quality attributes such as texture or rehydration
properties. While nano-CT (X-ray imaging at nanometer resolution) has recently become
available, the applicability of this new method remains to be explored. The aim of this work
was to visualize the 3D structure of selected moist and dry food products by advanced X-ray
imaging at micro- and nanometer resolution. In particular, we wanted to determine the
achievable representative sample size, resolution and contrast for imaging different types of
foods by quantitative comparison of images acquired at different spatial resolutions.

MATERIALS & METHODS

Sugar foams were produced in the lab by a standardized procedure (SGGW, Warsaw, Poland).
Apples (Malus domestica Borkh., cv `Braeburn') were picked on October 27th 2010 in an
orchard in Sint-Truiden (Belgium), and stored in Controlled Atmosphere (CA) coolrooms
(VCBT, Heverlee, Belgium). Nestlé and Chaber manufactured extruded cereal products by
using a model recipe of commercially available products. Extrusion conditions as well as
composition of the cereals could be modified. The samples were scanned using a SkyScan
1172 high resolution X-ray micro-CT system and/or the SkyScan 2011 nanotomograph
(SkyScan, Kontich, Belgium), operating at rather low energies ranging between 30 and 59 keV,
best suited for scanning these soft food materials. Reconstructed images, with pixel resolutions
at and below 1 μm, were processed using CTAn (SkyScan, Kontich, Belgium) and Avizo
(VSG, Bordeaux, France). In order to quantify the microstructures (Figure 1), CT images were

11th International Congress on Engineering and Food - Athens, Greece, 2011 2227
segmented by defining a threshold value separating different structures based on the grey-
scale, which correlates to the attenuation of the X-rays.

RESULTS & DISCUSSION

Projection images of the samples (Figure 1-top) show the presence of a microstructural features
in the foods, with contrasting elements such as high-density inclusions (black), strongly
attenuating the X-rays, and low-density regions (lighter) through which the X-ray beam passes
more easily. A more detailed insight in the microstructure is obtained by studying the virtual
cross-sections (Figure 1-bottom). Crispy breads show a highly connected bread matrix network
with air inclusions from μm to mm-sizes, leading to porosities as high as 91,79%. High density
inclusions can also be discerned (white). Reconstructed apple tissue images clearly show
clustered cells, surrounded by air voids, of critical importance for gas transport in the fruit.
Depending on the sample location inside the fruit, different porosities were measured: highest
porosities occur in the fruit cortex (20,18% ± 3, 28), followed by tissue under the skin (18,94% ±
1,19), whereas the least porous tissue is situated in the centre of the fruit (14,61% ± 3,4). The
foam was scanned at a range of resolutions, strongly affecting scan results. Increasing imaging
resolution causes smaller bubbles to be detected, resulting in a rise of measured porosity: from
1,63% at 70 μm pixel resolution to 64,45% at 1,35 μm pixel resolution. The cereal sample
scanned at 450 nm pixel resolution shows the presence of micro-cracks. Further morphometric
parameters such as local pore and structure diameters, connectivity and anisotropy were
obtained, which in combination with 3D visualisation (Figure 1E), provide a comprehensive
insight in the microstructure of these foods.

Figure 1. X-ray radiographic (top) and reconstructed (bottom) images of crispy bread (A), apple (B),
foam (C) and cereal (D). 3D visualization of segmented air bubbles in foam (E).

CONCLUSION
X-ray CT was very effective for imaging the microstructure of these porous products. The
distinct phases of the food (solid matrix, dense inclusions and air spaces) could be segmented
due to a high contrast in X-ray absorption. Nano-CT provided complementary structural
information (in particular the pore size distribution) to micro-CT but care had to be taken to
provide representative samples. Nevertheless, X-ray nano-CT enabled the investigation of the
3-D microstructure of samples in a near-native state at unprecedented resolutions. On a longer
term, this knowledge will contribute to improving nutritional quality (sugar- and gluten-free
cereal products), sensory quality (texture) and safety (foreign material detection) of foods.

2228
 Optical coherence tomography for quality control and microstructure analysis in food

Michael Leitnera,*, Günther Hannesschlägera, Attila Saghya, Alexandra Nemetha, Sophie Chassagne-
Bercesb, Hélène Chanvrierb, Els Herremansc, and Bert E. Verlindend

a
RECENDT – Research Center for Non Destructive Testing GmbH , Hafenstrasse 47-51, 4020 Linz,
Austria
b
NESTLE SA, Nestle PTC. Route de Chavornay 3, Orbe, Switzerland
c
Katholieke Universiteit Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium
d
VCBT, Flanders Centre of Postharvest Technology, Willem de Croylaan 42, B-3001 Leuven, Belgium
*Corresponding author electronic mail: michael.leitner@recendt.at

INTRODUCTION
Quality control and analysis of microstructure are of utmost importance in food industry. Pome
fruit, for example, are often stored for several months under special conditions, and the
thickness and homogeneity of the wax layer in the paring determines the apple´s protection
against liquid and therefore weight loss. In the case of extruded cereals the thickness and
homogeneity of sugar coatings, as well as the pore size distribution of the uncoated cereals, are
of special interest, since these determine the rehydration properties and the crisp- and
crunchiness, respectively. To control and monitor these quality indicators during the storage
and production processes of foods there is a need for fast and non-invasive assessment
techniques.
Optical coherence tomography (OCT) [1-4] is an emerging purely optical, non destructive, and
contactless high resolution imaging technique, which allows acquisition of two or three
dimensional image data in situ and in real time. OCT is the two and three dimensional
extension of low coherence interferometry and therefore well suited to image layered and
micro-structured specimens. The image contrast is due to inhomogeneities in the refractive
index of the sample materials, and thus OCT provides complementary information to other
high resolution imaging techniques, like X-ray computed tomography (CT) and magnetic
resonance imaging (MRI).

MATERIALS & METHODS

The experiments presented here were performed with two different OCT set-ups, which are
available at the labs of RECENDT, a time-domain ultra-high resolution (TD-UHR) OCT
system and a spectral-domain (SD) OCT system.
Braeburn apples were grown at an experimental station, Sint-Truiden, Belgium. Harvest date
was on 27/10/2010 which is in the optimal commercial picking window for long storage of
Braeburn in Belgium determined by Flanders Centre of Postharvest technology, Belgium. The
day after picking apples were sorted for size and kept at 1°C at normal air.

RESULTS & DISCUSSION

Pome fruit like apples are often stored for several months, and the quality and thickness of the
wax layer is one important parameter throughout the storing process [5]. To show the ability of
optical coherence tomography for the analysis and the control of wax layer thickness we

11th International Congress on Engineering and Food - Athens, Greece, 2011 2229
performed OCT imaging sessions on Braeburn apples. Figure 1 a) shows an OCT cross-section
image, as acquired with the SD-OCT system. The image size is 4 x 1.25 mm² and several
layers of the paring can clearly be distinguished. Other interesting features for the storage life
of apples are the lenticels, which act as a bypass medium for the exchange of gases between the
fruit flesh and the ambient. However, also bacteria and funguses can penetrate the fruit through
the lenticels. Panel 1 b) shows an OCT image of a lenticel, as acquired with the TD-UHR-OCT
set-up, and depicts a cross section with an image size of 3 x 0.3 mm². The lenticel is clearly
visible in the lateral centre of the image, as indicated by the arrow. The full paper also
describes OCT applications for microstructure analysis in extruded breakfast cereals.

Figure 1: OCT images of Braeburn apples; a) Cross section image acquired with the SD-OCT system.
Image size: 4 x 1.25 mm²; b) Cross-section image of a lenticel, acquired with the TD-UHR OCT system.
Image size: 3 x 0.3 mm²;

CONCLUSION
In this work we introduced optical coherence tomography as a new tool for microstructure
analysis and quality control in food. We showed the capability of this fast and non invasive
optical imaging technique for a real time assessment and monitoring of microstructures, and
therefore as a promising tool for at line quality control of food.

ACKNOWLEDGEMENTS: This work has been carried out with financial support from InsideFood -
Integrated sensing and imaging devices for designing, monitoring and controlling microstructure of foods
(FP7-226783).

REFERENCES
[1] Huang D, Swanson EA, Lin CP, Schuman JS, Stinson WG, Chang W, Hee MR, Flotte T, Gregory K, Puliafito CA,
Fujimoto JG 1991. Optical Coherence Tomography. Science 254(5035):1178-1181. [2] Stifter D 2007. Beyond
biomedicine: a review of alternative applications and developments for optical coherence tomography. Applied Physics
B: Lasers and Optics 88(3):337-357. [3] Wiesauer K, Pircher M, Götzinger E, Hitzenberger CK, Oster R, Stifter D
2007. Investigation of glass-fibre reinforced polymers by polarisation-sensitive, ultra-high resolution optical coherence
tomography: Internal structures, defects and stress. Composites Science and Technology 67(15-16):3051-3058. [4]
Wiesauer K, Pircher M, Gotzinger E, Bauer S, Engelke R, Ahrens G, Grutzner G, Hitzenberger CK, Stifter D 2005.
En-face scanning optical coherence tomography with ultra-high resolution for material investigation. Optics Express
13(3):1015-1024. [5] E. A. Veraverbeke, N. Van Bruaene, P. Van Oostveldt, and B. M. Nicolaï, Non destructive
analysis of the wax layer of Apple (Malus domestica Borkh.) by means of confocal laser scanning microscopy. Planta
213(4), 525-533 (2001).

2230
 Effect of Fibres and Whole Grain Content on Quality Attributes of Extruded Cereals

Sophie Chassagne-Bercesa, Michael Leitnerb, Angela Meladoc, Pila Barreiroc, Eva Crostina Correac, Imre
Blanka, Jean-Claude Gumya, Hélène Chanvriera

a
NESTEC SA, Nestle PTC Orbe, 1350 Orbe, Switzerland (sophie.chassagne@rdor.nestle.com,
helene.chanvrier@rdor.nestle.com)
b
RECENDT, 4020 Linz, Austria (michael.leitner@recendt.at)
c
UPM, 28040 Madrid, Spain (angela.melado@upm.es, pilar.barreiro@upm.es)

INTRODUCTION
Health and nutritional policies are currently promoting the increase of dietary fibre content in
food, especially in cereal-based products. However, incorporation of fibre in cereals may lead
to quality issues [1-2], thus decreasing consumer acceptance. This is partially due to
deterioration of the microstructure, one of the primary quality attributes of cereals [3-5].
Consequently, the production of fibre-enriched extruded cereals remains a challenge, in
particular when maintaining functional and quality properties.
The objective of this study was to better understand the mechanisms by which dietary fibres
affect the quality of cereal products during extrusion-cooking, by quantifying the effect of
source and amount of fibre and whole grain on (i) texture, (ii) structure, and (iii) rehydration
properties of extruded cereals. New innovative methods were applied and combined with
traditional techniques to characterize both the structure and the rehydration properties.

MATERIALS & METHODS

Studies were carried out on starch-based (wheat, whole wheat) recipes. Two sources of fibres
were added: oat bran concentrate and wheat bran for their high soluble (ȕ-glucans) and
insoluble (arabinoxylans) fibre levels, respectively. The oat and wheat bran levels used in this
study were 0, 10, 20%. The different recipes were extruded in a pilot twin-screw extruder
BC21 (Clextral) and then sugar coated after drying. The following extrusion parameters were
kept constant: die design, screw speed (400 rpm), product temperature (135°C) and moisture
content (20%).
Mechanical properties of extruded cereals were investigated by compression test. The cellular
structure was observed by X-ray tomography. Information on porosity, cell size and cell wall
thickness distributions were extracted from 3D image analysis. The quality of coating
(thickness, homogeneity) was analysed by optical coherence tomography. The rehydration
properties of the extruded cereals in milk were evaluated by magnetic resonance imaging
(MRI) and optical coherence tomography.

RESULTS & DISCUSSION

Whatever the type of fibre (oat bran concentrate or wheat bran), the modifications of
mechanical properties after addition of fibres or whole grains are similar:
x Without addition of fibres (0%), the maximum force and the number of peak do not
significantly change when the whole grain content increases.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2231
x Conversely, adding fibres increases significantly the maximum force (Fmax), whereas the
number of peak (Npeak) decreases, thus showing an increase of hardness and a decrease of
“crispness”, when the fibres are added.
The modifications of texture parameters (Fmax and Npeak) seem to be more important with oat
bran concentrate than with wheat bran.

Modifications of mechanical properties were linked with variations of cell size and cell walls
structure.
x Without addition of fibres, no modification of porosity and the cell size is observed when
the whole grain content increases.
x Conversely, adding fibres decreases the expansion of extruded cereals and thus the cell
size and the porosity decreases while the thickness of cell wall increases.

A loss of hardness and crispness is observed after immersion in milk. The hardness of dry
products is well correlated with those of the soaked products, thus showing the effect of fibres
addition on “keeping hardness” when poured in milk. This is confirmed by MRI measurements
showing a slower penetration of the milk within the pellets containing fibres.
OCT made possible to visualize the quality of coating and to follow the rehydration process of
extruded cereals in milk: the collapse of the structure that is immerged in milk can be followed
and quantified. Furthermore, MRI reveals that the differences of rehydration properties
between coated and non coated extruded cereals seem to be dependent on the content and
composition of the cereal base.

CONCLUSION
This work revealed that structure assessment of extruded cereals may lead to a better
understanding of the effect of fibre addition on texture and rehydration properties. The
application of innovative methods, such as Optical Coherence Tomography and Magnetic
Resonance Imaging, was found to be useful to quantify the structural properties. In the future,
the relationships between quantitative analysis of expanded structure changes in extruded
cereals with high fibre content and the final texture properties will be used to define optimized
processing conditions and recipe for an improved consumer satisfaction.

REFERENCES
[1] Yanniotis S., Petraki A., Soumpasi E. (2007). Effect of pectin and wheat fibers on quality attributes
of extruded cornstarch. Journal of Food Engineering, 80(2), 594-599.
[2] Brennan M. A., Merts I., Monro J., Woolnough J. & Brennan C.S. (2008). Impact of Guar and Wheat
Bran on the Physical and Nutritional Quality of Extruded Breakfast Cereals. Starch – Stärke, 60(5),
248-256.
[3] Jin Z., Hsieh F. & Huff H.E. (1995). Effects of soy fiber, salt, sugar and screw speed on physical
properties and microstructure of corn meal extrudate. Journal of Cereal Science, 22(2), 185-194.
[4] Yao N., Jannink J.L., Alavi S. & White P.J. (2006). Physical and sensory characteristics of extruded
products made from two oat lines with different ȕ-glucan concentrations. Cereal Chemistry, 83(6),
692-699.
[5] Rzedzicki Z. & Blaszczak W. (2005). Impact of microstructure in modelling physical properties of
cereal extrudates. International Agrophysics, 19(2), 175-186.

2232
NMR microscopy and NMR HR-MAS on apples of different qualities after different

storage conditions

Dieter Grossa, Manfred Spraula, E. Humpfer, H. Schaefer, A. Melado, T. Defraeye, P. Verboven

a
Bruker Biospin, Germany

A Digital Laboratory for visual analysis of materials microstructure

Pascal Estrade

VSG,France


Cryo scanning electron microscopy: enabling nano-imaging of food products

Frederic Depyperea, D. Van de Walle, K. Dewettinck

a
UGent, Belgium

11th International Congress on Engineering and Food - Athens, Greece, 2011 2233
2234
 The Application of Acoustic Emission to Measure Texture of Food Foams

Ewa Jakubczyk, Ewa Gondek

a
Department of Food Engineering and Process Management, Warsaw University of Life Sciences,
Nowoursynowska 159C, 02-776 Warsaw, Poland, ewa_jakubczyk@sggw.pl, ewa_gondek@sggw.pl

INTRODUCTION
Bubbles are integral to many classic confectionery products such as marshmallow, nougat, and
meringue [2]. Candy-foam like structures can be obtained by blowing gas through a nozzle to
the product or mechanical agitation of sugar-agar-foaming agent solutions [1].
The texture measurement of candy foams was presented by a typical force-deformation
response for aerated material in tension [2]. The mechanical parameters of an aerated gel
system were also analysed using a compression test.
Acoustic emission is a method that enables characterization of texture of the materials because
the sound wave contains information about the microstructure and micromechanical properties
of the product. In acoustic emission (AE) the sound wave produced during mechanical
deformation of an object such as a food is captured using a sensor. Acoustic emission was
successfully used to measure the differences in texture of different products [3].
The aim of this work was to evaluate the possibilities of applying the acoustic emission method
to determine the texture of aerated food gels.

MATERIALS & METHODS

The agar-fructose solution with addition of albumin from chicken egg was foamed using a
kitchen mixer. The effects of different whipping time on mechanical and acoustic properties
were determined. The solution was aerated for 1, 3, 5, 8, 10 and 15 minutes. Apparent density
of foams was calculated based on mass of aerated gel divided by occupied volume.
The measurement of acoustic emission was carried out while compressing prepared samples
diced into 13 mm cubes. Aerated gels were compressed with a speed of 50 mm/min. Acoustic
emission was registered in the range 0.1-16 kHz using a piezoelectric accelerometer type
(Bruel & Kjaer). The recorded AE signal was amplified in the external low noise amplifier and
recorded. The selected acoustic descriptors were analysed.

RESULTS & DISCUSSION

The aeration of sugar-agar solution affected the density and acoustic properties of gelled
material. The density of aerated gel after 1 minute of whipping was reduced by about 34% in
comparison to non-foamed agar gel. The increase of whipping time from 1 to 3 minutes caused
a significant decrease of sample density (Table 1). It is evident that the density of foams
slightly decreased with whipping time. However, there was a marked increase in density
observed between 10 and 15 minutes. Overbeating may lead to damage of the structure of
aerated material.
The texture of aerated gels was also measured using the acoustic emission method (Table 1).
There was a significant increase in the number of acoustic events observed between 1 and 10
minutes. Longer aeration resulted in a decrease of the number of acoustic events and average
energy of a single acoustic event. The total acoustic energy of the material gradually increased

11th International Congress on Engineering and Food - Athens, Greece, 2011 2235
with whipping time. The porous structure of gels had a significant effect on acoustic emission
of the gel system. Damage to many pores during compression may lead to emission of an
acoustic signal. The decrease of number of acoustic events and acoustic energy of single events
and after 15 minutes of whipping may indicate changes in bubble size and in distribution of air
voids in the material.

Table 1. Acoustic descriptor and apparent density of foams after different whipping times
Whipping time Apparent density Acoustic energy of Number of Total acoustic energy
(min) (g/cm3) single event (mV) acoustic events (a.u.)
1 0.839 r 0.020 142.42 33.65 96.78 r 3.61
3 0.523 r 0.001 307.65 78.65 107.68r 2.20
5 0.469 r 0.001 249.10 149,95 131.48 r 7.59
8 0.490 r 0.009 246.65 175.20 130.92 r 8.19
10 0.460 r 0.009 260.19 210.25 148.33 r 8.07
15 0.482 r 0.001 206.35 198.00 150.90 r 8.25

The decrease of density after aeration was a result of air incorporation. The results of many
authors have shown that with whipping time the size of bubbles decreased but the number of
bubbles progressively increased. The more acoustic events observed with longer whipping time
indicated that more bubbles were deformed during compression. The decline in the number of
acoustic events may be a result of higher density of foams probably caused by overbeating.

CONCLUSION
1. The density of sugar-gel foam decreased with whipping time (1-5 min). The increase in
density observed between 10 and 15 minutes may be a result of structural changes and
agglomeration of bubbles.
2. The foam structure of sugar gels had a significant effect on acoustic emission of the gel
system. Damage to pores during compression may lead to emission of an acoustic signal.
3. A higher total number of acoustic events indicated a larger number of bubbles deformed
during compression.

ACKNOWLEDGEMENTS
This study was funded within the framework of the European project InsideFood (FP7-226783
‘Integrated sensing and imaging devices for designing, monitoring and controlling
microstructure of foods’).

REFERENCES
[1] Campbell G. M. & Mougeot E. 1999. Creation and Characterisation of Aerated Food Products. Trends
in Food Science and Technology, 10, 283-296.
[2] Decker N. R. & Ziegler G. R. 2003. Mechanical Properties of Aerated Confectionery. Journal of
Texture Studies, 34, 437-448.
[3] Lewicki P. P., Marzec A. & Ranachowski Z. 2009. Acoustic Properties of Foods. In: Rahman S. M.
(Ed.). Food Properties Handbook. 2nd Ed. CRC Press Taylor & Francis Group., Boca Raton, USA.

2236
 Non destructive detection of brown heart in ‘Braeburn’ apples by time-resolved

reflectance spectroscopy

M. Vanolia,b, A. Rizzoloa, M. Grassia, A. Farinab, A. Pifferib, L. Spinellic, B. E. Verlindend, A. Torricellib

a
CRA-IAA, Milan, Italy (maristella.vanoli@entecra.it)
b
Politecnico di Milano, Dipartimento di Fisica, Milan, Italy (alessandro.torricelli@polimi.it)
c
Istituto di Fotonica e Nanotecnologie – CNR, Milan, Italy (lorenzo.spinelli@fisi.polimi.it)
d
Flanders Centre of Postharvest Technology (VCBT), Leuven, Belgium
(Bert.Verlinden@biw.kuleuven.be)
INTRODUCTION
Brown Heart (BH) is an internal disorder related to CO2 injury, which is characterized by browning
of the pulp and formation of cavities, and it is visible only when fruit are cut open. The
susceptibility of Braeburn apples to BH is related to their structural characteristics, as they have a
relative dense and firm tissue, poor flesh gas diffusivity and low skin gas-permeance. The
unpleasant nature of BH is not acceptable to consumers and causes economic losses. As external
symptoms are not evident, a reliable non-destructive method for on-line detecting and segregating
damaged from healthy fruit would be readily accepted by large co-operatives and commercial
packing-houses. Previous studies have shown that time-resolved reflectance spectroscopy (TRS) is
able to detect BH in pears, internal browning in Granny Smith apples, watercore in Fuji apples and
mealiness in Braeburn apples, showing higher absorption coefficients in the range 720-850 nm in
fruit affected by disorders [1]. In the present work, the optical properties measured by TRS were
evaluated in order to test whether TRS can be used to detect BH in intact “Braeburn” apples.

MATERIALS & METHODS

‘Braeburn’ apples, picked at commercial harvest in Belgium, were stored at 1°C for 3 and 6 months
in BH inducing (1% O2 + 5% CO2, BH storage) and not-inducing (2.5% O2 + 0.7% CO2 with a 3
week delay of CA, OPT storage) conditions. At each storage time, sixty apples/storage were
measured by a broadband TRS setup [2] at 670 nm and in the spectral range 740-1100 nm on four
points (A-D) around the equator, ranked on the basis of decreasing μa670 (increasing maturity) and
divided into 2 batches correspondent to 2 times of shelf life at 18°C (day 0 and day 14). At d14
apples were measured by TRS in the same spectral range and points as at d0. Afterwards, each fruit
was cut open and evaluated for disorders (browning, cavity) recording the position (brown core, BC;
brown pulp, BP) and the association with cavities (browning alone, BA; browning plus cavities,
BCV), and the severity score (1=healthy, 2=very slight, 3=slight, 4=moderate and 5=severe). All
fruit were measured for flesh firmness, intercellular space volume (RISV) and pulp colour (L*, a*,
b*, C* and H°, spectrophotometer CM-2600d, at 18 mm from the skin in correspondence of A-D
points of TRS measurements). The average of all points/fruit were computed before submitting
optical and colour data to ANOVA. Correlations between optical and colour data were studied using
the PROC CORR procedure.

RESULTS & DISCUSSION

Internal browning and cavities were already present after 3 m in BH storage, while in OPT storage
no disorder was found. After 6 m and at the end of shelf life, in BH storage there was the highest

11th International Congress on Engineering and Food - Athens, Greece, 2011 2237
incidence of browning (90%) and cavities (42%), while in OPT storage the browning incidence was
55% and the cavity ones 7%. Browning was localized mainly in the core region in both
atmospheres; the incidence of BP increased with increasing shelf life time, mainly in BH storage
after 3 m and in OPT storage after 6 m.
Overall, fruit affected by internal browning showed significantly higher μa in the 740-900 nm
spectral range respect to those of healthy ones, with the highest difference recorded at 740 nm. With
the development of internal browning, μa740 increased, more in BP than in BC fruits, and more in
BCV than BA ones (Table 1). There was no difference in μa740 between pulp without browning and
pulp affected by very slight and slight browning, whereas it significantly increased in fruit with
moderate and severely affected fruit. L* and a* increased and H° decreased in fruit with moderate
and severe browning. Firmness did not change with respect to browning score, browning position
and cavities, whereas RISV showed the lowest percentage in healthy apples and the highest in
browned fruit scored as moderate and severe, and when cavities were associated to browning. As
the TRS measurement points did not always corresponded to the browning area in the pulp, in order
to study the correlations between μa740 and colour parameters, for each browned fruit only the
measurement points in which TRS analysis was carried out in correspondence of the defect were
chosen. High correlations were found between μa740 and L* (r=í0.95), a* (r=0.88) and H°
(r=í0.88). From the correlation between μa740 and pulp L* at μa740 <0.038 cmí1 only healthy pulp
can be detected, and at μa740>0.08 cmí1 only severely browned pulp can be found.

Table 1. Mean values of μa740 and of pulp colour parameters in healthy and browned Braeburn apples
(brown core, BC; brown pulp, BP; brown fruit without cavities, BA; brown fruit with cavities=BCV).
μa740 (cm- L* a* b* C* H°
1
)
Healthy 0.033 c C 82.83 a A í0.40 b B 23.30 b A 23.34 b A 90.98 a A
BC 0.037 b 81.52 b í0.21 b 23.12 b 23.18 b 90.56 a
BP 0.064 a 74.08 c 2.92 a 24.51 a 24.84 a 83.44 b
BA 0.040 B 80.81 B 0.09 B 23.23 A 23.33 A 89.86 A
BCV 0.047 A 78.35 C 1.10 A 23.76 A 23.90 A 87.60 B
small letters refer to browning position, capital letters refer to browning and cavities

CONCLUSION
The absorption coefficient measured at 740 nm was able to segregate healthy fruit from those
having moderate and severe BH. However, when browning affects only the region near the pith,
TRS was not able to detect the disorder due to its depth, which was greater than 2cm. Furthermore,
when internal browning involves only part of the pulp, the four equidistant measurement points used
in this research were not enough to make certain the detection of the disorder.

REFERENCES
[1] Vanoli M., Rizzolo A., Eccher Zerbini P., Spinelli L. & Torricelli A. 2009. Non-destructive detection of
internal defects in apple fruit by Time-resolved Reflectance Spectroscopy. Int. Conf. COST 924
“Enviromentally Friendly and Safe Technologies for Quality of Fruits and Vegetables”, Faro, Portugal, 14-16
January, 2009, in press.
[2] D’Andrea C., Nevin A., Farina A., Bassi A. & Cubeddu R. 2009. Assessment of variations in moisture content
of wood using time-resolved diffuse optical spectroscopy. Applied Optics, 48(4), B87-B9.

2238
Non-destructive Characterization of Food Microstructure and Composition by Spatially-

Resolved Spectroscopy

N. Nguyen Do Tronga, M. Tsutaa, b, E. Herremansa, R. Wattéa, C. Erkinbaeva, E. Verhoelsta,

P. Verbovena, B. M. Nicolaïa, W. Saeysa
a
Division of Mechatronics, Biostatistics and Sensors (MeBioS), Department of Biosystems, K.U.Leuven,
Kasteelpark Arenberg 30, 3001 Leuven, Belgium.
Email: nghia.nguyendotrong@biw.kuleuven.be
b
National Food Research Institute, 2-1-12 Kan-nondai, Tsukuba, Ibaraki 305-8642, Japan

INTRODUCTION
Quality of foods strongly depends on their microstructure and composition. Examples include
sponginess of bread, crispness or crunchiness of crackers, firmness or sweetness of fruits.
Processing of foods also affects their microstructure and composition: existing structures are
destroyed and new ones are formed; some constituents are changed and new ones are created.
Therefore, rapid and accurate measurement of food microstructure and composition and how
they change during processing operations is essential for the production of high quality foods.
Microstructure and composition (and their changes) strongly determine light propagation
behavior (e.g. diffuse reflectance) in the illuminated food samples, mostly attributed by
scattering and absorption phenomena. However, multiple light scattering increases photon
pathlengths inside the biological structure resulting in increased absorbance for the same
concentration level, such that the measured reflectance or transmittance spectra result from the
interplay of both scattering and absorption. Since the traditional NIR spectroscopy technique
only measures the diffuse reflectance (or transmission), which is a combination of scattering
and absorption effects, further improvements could be implemented for resolving this problem.
This research was performed in the context of the EU project InsideFood (FP7-226783). In this
study, the potential of spatially resolved spectroscopy for non-invasively characterizing
microstructure and composition of the microstructured foods (model foods) by means of their
optical properties (absorption and reduced scattering coefficients) has therefore been
investigated.
MATERIALS & METHODS
A setup for SRS measurements in the 400-1100 nm range has been built. This setup consists of
a contact probe with accurately placed fibers which is linked to a spectrograph for
simultaneous measurement of the reflectance at the different distances by a CCD camera. The
optical probe has been designed and assembled at the Swiss Federal Institute of Technology
(EPFL, Lausanne, Switzerland). The fibers used are Thorlabs multimode silica fibers (FVP-200
PF) with a numerical aperture of 0.22 and a core diameter of 200 Pm. The 7 detection fibers
are placed at various distances from the illumination fiber, ranging approximately from 0.3 to
1.2 mm with a step of about 0.15 mm. The illumination fiber of the probe is connected to a
AvaLight-DHc (Avantes, Eerbeek, The Netherlands) halogen lamp through an optical switch.
The detection fibers from the SRS probe and a fiber from the optical switch of the light source
are aligned in the entrance slit of a CP200 133 g/mm spectrograph (Horiba Jobin-Yvon, New
Jersey, USA) which splits the light from each of these fibers into its spectral components and
projects these onto a Hamamatsu C7041 CCD camera with a S7041-1008 detector

11th International Congress on Engineering and Food - Athens, Greece, 2011 2239
(Hamamatsu, Louvain-La-Neuve, Belgium). The signal from this camera is transferred to a
computer by means of a PCI MIO-16E-4 data acquisition card. Control of the light source,
optical switch and camera is performed in LabView software (National instruments, TX, USA)
Several model foods with clearly different microstructure properties have been designed. A
first food model is made by mixing agarose and gelatin in water to create an emulsion or a gel.
Two mixtures are considered: 1% agarose with 1% gelatin (Gel 1) and 1% agarose with 2.5%
gelatin (Gel 2). A second model food are the candy foams. These are produced by mixing
fructose, dextrose agar-agar, albumin and water. Two mixtures have been considered in this
study: One without dextrose (Foam 1) and one with dextrose (Foam 2). The third model food
considered in this study is a chocolate mousse which was created by mixing cold swelling
starch with cocoa, sugar, oil and water.

RESULTS & DISCUSSION

The optical properties of the different model foods are then estimated:

Gel 1 Gel 2 Foam 1 Foam 2 Chocolate Mousse

Figure 1. Fitted μs’ (upper left) and fitted μa (upper right) of 5 model foods. The continuous and dotted
lines represent the mean and 95% confidence intervals of the fitted values respectively. (Bottom)
Microstructure images of the model foods by microscopy (the scales in the images are 10 μm).

CONCLUSION
A spatially-resolved spectroscopy setup based on a fiber-optic probe was successfully
elaborated in the lab and validated for its measurement accuracy. A logical correlation was
found between the estimated reduced scattering coefficient spectra and the designed
microstructures of these model foods, verified by light microscopy. The estimated absorption
coefficients also showed good agreement with the designed ingredients. This research clearly
indicates the potential of spatially-resolved spectroscopy methods for non-invasive food quality
inspection and process monitoring in the food industry.

2240
 New Tools, concepts and solutions for improving technologies along the

European food cold chain: the FRISBEE project

Graciela Alvareza, A. Geeraerd b, D. Leducq a, J.Evans c, E. Wissink d, E. Indergårde

C. Cotillonf, P. Taoukis g

a
CEMAGREF, France (graciela.alvarez@cemagref.fr)
b
BIOSYST-MeBioS, K.U.Leuven, Leuven, Belgium c LSBU, Langford, UK,d TNO, Apeldoorn, The
Netherlands,eSINTEF Energy Research, Norwayf ,ACTIA, , France
g
Chemical Engineering Department, NTUA, Athens, Greece

INTRODUCTION
FRISBEE is a Food Refrigeration Innovation for Cold Chain Research IP European project.
The four-year, 6 M euro project is funded mainly through the EU’s 7th Framework
Programme, and has 26 partners; 13 of which are companies, 11 research institutes or
universities, and 2 nongovernmental organisations. FRISBEE will provide new tools, concepts
and solutions for improving refrigeration technologies along the European food cold chain.
The project will develop new innovative mathematical modelling tools that combine food
quality and safety together with energy, environmental and economic aspects to predict and to
control food quality and safety in the cold chain. Many innovative and high-tech food
refrigeration technologies such as advanced control, thermal storage, nanoparticles, super
chilling and magnetic refrigerator will be studied. Disruptive new technology will be
developed for a new energy-efficient refrigerator which would use magnetic refrigeration
instead of the vapour-compression cycle currently universally used.

FRISBEE main objectives and expected results

Frisbee will develop, first, a comprehensive database of the cold chain in Europe, identifying
refrigeration needs and available current technologies in the food industry, and investigating
consumer needs and expectations with respect to the food cold chain. We will compile product
temperatures existing data in Europe.
The project will establish the development of novel Quality and Energy/Environment
assessment Tools: The QEEAT. These tools are models, sensors, equipment, protocols and
methodologies that combine food quality and safety together with energy, environmental and
economic aspects to manage food quality and safety in the cold chain and energy optimisation.
These tools will allow assessing and improving existing refrigeration technologies and
emerging new technologies. Particular effort will be performed on:
a) Quality models Considerable efforts have been made in the last twenty years to develop
mathematical models to predict quality attributes of refrigerated foods. These attributes can be
referring to microbial quality, and physicochemical quality attributes. Obviously, temperature
is one of the most important controlling factors within this context. Most research has been
performed under stationary temperature conditions to obtain model parameters of the quality

11th International Congress on Engineering and Food - Athens, Greece, 2011 2241
degradation kinetics. By recording temperature conditions of the product continuously by a
temperature sensor, (e.g., inexpensive robust time–temperature sensors), consistent estimation
of the quality status and the remaining shelf life of the products could be achieved based on
models for quality tracers. Quality models under variable conditions will be developed for
FRISBEE.
b) Stochastic aspects of the cold chain Combining thermal deterministic and stochastic models
approaches has to be studied in food process engineering due to the variation of product
biological properties under uncertain process conditions. The main goal for these studies is the
optimization of cost, product quality and safety. Different methods have been proposed in these
studies to quantify the effects of the uncertainty of model parameters on the output of the
studied system. One widely used method is the Monte Carlo, which requires a large number of
repetitive simulations to obtain an acceptable level of accuracy.
Very few research work has been performed to take into account stochastic aspects during cold
storage, although the product is often exposed to uncertain environmental conditions such as
ambient temperature for variable, product position and durations in refrigerating equipment.
The methodology to be developed needs to focus on the prediction of the evolution and
variability of product temperature, microbial load and total energy needed for conservation
along the cold chain.
c)Energy use quality and sustainability .Advanced predictive control of refrigerating plants
Worldwide, refrigeration consumes 8 % of all energy and is responsible for 2.5 % of
greenhouse gas emissions, therefore any reduction to those figures will be a big improvement.
To improve existing technologies we will use new concepts such thermal energy storage
devices, nanoparticules of phase change materials (PCM) together with new advanced control
methods. Multiobjective optimisation approach will be performed to maintain food quality
reduce energy consumption, environmental impact in refrigeration process.
Model predictive control (MPC) will be developed in FRISBEE to predict the future response
of the system taking into account scenarios such as energy availability and price, weather
forecast, provisional load of the production line. We will take into account several difficulties
such as interaction between nonlinear dynamics and discrete events, on/off manipulated
variables, continuous controlled variables such as temperatures set points and finally, several
operation constraints.
d) Emerging new refrigeration technologies
If we will be able to develop such QEEAT Quality Energy, Environment Tools, and advanced
control algorithm the next step will be to develop new emerging refrigeration technologies,
such as nanofluids, air refrigerating machine or magnetic refrigeration. We will be able to
develop as well new refrigeration processes such as superchilling and supercooling and finally
to transfer and disseminate these innovations to all sector in the cold chain in particular to end-
users such as: industrial and stakeholders and consumers associations

REFERENCES
http://www.frisbee-project.eu

ACKNOWLEDGEMENT
The research leading to these results has received funding from the European Community‘s
Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 245288.

2242
 Management and Optimization of the Cold Chain and the development of

Cold Chain Data Base

Petros Taoukis, George Katsaros, T. Tsironi, E. Dermesonluoglu, E. Gogou

Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical
University of Athens, Greece, (taoukis@chemeng.ntua.gr)

INTRODUCTION
The main shelf-life determining post-processing parameter in the cold chain of chilled and
frozen food products is temperature. A modern quality and safety assurance system should rely
on prevention through monitoring, recording and controlling of critical parameters during the
entire product’s life cycle that includes the post-processing phase and extends to the time of
use by the final consumer. Increasing attention is focused on the role and the logistics of
transport, storage and handling, and the benefits of taking a supply chain perspective are being
appreciated and pursued. Temperature conditions in the chilled distribution chain determine the
risk potential, the shelf life and final quality of chilled products processed and packed under
Good Manufacturing Practices and Good Hygiene Practices. Since in practice significant
deviations from specified conditions often occur, temperature monitoring and recording is a
prerequisite for chain control and any logistics management system that aims on product
quality optimisation at the consumer’s end [1, 2].

MATERIALS & METHODS

A systematic data collection for identification and evaluation of the weak links of the cold
chain for different types of chilled and frozen products is necessary. A web-based platform
(hosted in the link http://frisbee-wp2.chemeng.ntua.gr/) has been built for data collection,
maximizing information retrieval with user friendliness. At all stages of the cold chain, the
needs of consumer and European industry will be considered, gaining a greater insight into
deviations between real cold chain data and targeted specifications.

RESULTS & DISCUSSION

Data from industry, cold chain parties (distributors, retailers) and consumer surveys, including
all stages of the cold chain (from production to consumption) are collected. All contributors
have privileged access to this database (by login and password) and the access to the database
is secured. This platform consists of a menu driven web-based software retrieving information
to accompany the contributed food product time-Temperature data. Some of the most
important questions to be answered from the user include the stages of the cold chain (i.e.
production warehouse, transportation etc.), the country of origin and the destination country of
the products and some descriptive information of the product such as the food storage
temperature range (chilled, frozen etc.), the characterization (fresh unprocessed, processed
ready to eat etc.) and type of food (meat and meat product, vegetable etc.), the packaging (air
packaged, vacuum packaged etc.) and the recommended food storage conditions. Some details
are asked with regards the data collecting equipment, such as the type and accuracy, date of last

11th International Congress on Engineering and Food - Athens, Greece, 2011 2243
Add Cold Chain Data 0 calibration and position during temperature
Stage/step of cold chain
Required Field Production warehouse
recording (top of the food, below the food
Transportation etc.). During on-line data submission, it may
Distribution warehouse be mentioned whether the data are
Retail warehouse
confidential or not. Any input will be
Hypermarket
Supermarket
valuable in building a comprehensive and
Hard discounter extensive database which will serve as a
Grocery valuable tool to people and organizations
Retail display
that have contributed and are involved in the
Consumer domestic refrigerator
Complete cold chain
Cold Chain as researchers or industrial
Other players. To enter the platform the user has to
login after creating his own account. After
Country of origin Please select an item
entering in the platform, a screen with the
Required Field
uploaded files of the user is shown (in the
Destination country Please select an item
first login there are no files, since no file has
been uploaded yet). To upload a file the user
has to click on the “Add Cold Chain data”
Sample Date 15/02/2011
button and he is driven to a new screen
Time data logger started
collecting data (hrs:min)
Please select an item
where all the necessary information
(metadata) for the uploaded time-
Food storage temperature
range
Please select an item
temperature profiles are shown. After filling
Characterization of food Please select an item in the available information in the boxes, he
Required Field
can upload the file with the time-temperature
Type of food Please select an item
data (in .xls format or any other format
available). After filling in the boxes and
selecting the file for uploading, he has to
click on the "Apply" button. Then, the software drives him to the first screen where now he
may see the uploaded file and all its metadata (double-click on the file).

CONCLUSION
The developed FRISBEE cold chain web based platform offers the potential to effectively
manage and improve cold chain weak links. The platform will offer concepts and solutions for
improving refrigeration technologies along the European food cold chain. The contributed data
of the cold chain will allow one to run simulations and distribution scenarios based on real cold
chain data. Further information with regards the web based platform and the data input
procedure may be provided by contacting frisbee@chemeng.ntua.gr.

REFERENCES
[1] Giannakourou M & Taoukis P. 2003. Application of a TTI-based distribution management system
for quality optimization of frozen vegetables at the consumer end. Journal of Food Science, 68(1),
201-209.
[2] Koutsoumanis, K., Taoukis, P.S., Nychas, G.J.E. 2005. Development of a Safety Monitoring and
Assurance System for chilled food products. Int. Journal of Food Microbiology, 100(1-3), 253-260.
ACKNOWLEDGEMENT
The research leading to these results has received funding from the European Community‘s
Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 245288.

2244
 Towards a framework for evaluation of energy consumption, sustainability and

associated food quality in the European cold chain

Sunny George Gwanpuaa, Bert Verlindenb, Sietze van der Sluisc, Edo Wessinkd,
Judith Evanse, Tim Browne, Denis Leducqf, Graciela Alvarezf, Petros Taoukisg, George Katsarosg,
Valérie Stahlh, Dominique Thuaulti, Ingrid Claussenj, Erlend Indergårdj,
Pieter Verbovena, Bart Nicolaïa, Annemie Geeraerda

a
BIOSYST-MeBioS, K.U.Leuven, Leuven, Belgium (annemie.geeraerd@biw.kuleuven.be)
b
VCBT, Leuven, Belgium (bert.verlinden@biw.kuleuven.be)
c
SAINT TROFEE, Renesse, The Netherlands (s.m.vandersluis@gmail.com)
d
TNO, Apeldoorn, The Netherlands (edo.wissink@tno.nl)
e
LSBU, Langford, UK (j.a.evans@lsbu.ac.uk)
f
CEMAGREF, Antony, France (graciela.alvarez@cemagref.fr)
g
Chemical Engineering Department, NTUA, Athens, Greece (taoukis@chemeng.ntua.gr)
h
Aérial, ACTIA Centre, Illkirch, France (v.stahl@aerial-crt.com)
i
ADRIA, ACTIA Centre, Quimper, France (dominique.thuault@adria.tm.fr)
j
SINTEF Energy Research, Trondheim, Norway (Ingrid.C.Claussen@sintef.no)

INTRODUCTION
Many models have been developed to explain temperature evolution and resulting food product quality and
safety along cold chains, but they have not yet been combined into a user-friendly software. Moreover, although
refrigeration is very important in extending the shelf life of perishable products, it has a setback of being a
major user of energy and a contributor to global warming. Reducing energy usage by refrigeration will
contribute in attaining the EU Commission’s objective of reducing energy consumption by 20% by 2020.
Models for refrigeration cycles can be used to predict the energy usage. Also, quantifying CO2 emission is
important in order to quantify the impact of a refrigeration technology on the environment. This FRISBEE
presentation focuses on a framework that is currently being developed to evaluate energy consumption,
environmental impact and associated food quality and safety attributes in the European cold chain.

MATERIALS & METHODS

As a first step, a reference product needs to be chosen for different product categories and the most important
quality and safety indicators for the different products are selected. For each reference product, both physical
(e.g. mass, geometry, diameter) and thermophysical properties (e.g. density, heat capacity, thermal
conductivity) are being defined. A reference cold chain, starting from when the food is
harvested/catched/slaughtered, to when it reaches the final consumer is currently being defined and quantified
for each product. Further elements of the framework are described below.

RESULTS & DISCUSSION

Reference food products, quality and safety indicators & reference cold chains
The reference food products were selected based on their economic importance in the EU market and are listed
in Table 1.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2245
 Table 1. Safety and quality indicators for selected food products & start of the reference cold chains.
Category Reference Safety indicator Quality indicator Start of the reference
food product cold chain
Chilled
Fruit Apple - Firmness, colour, Batch cooling of box
aroma pallets after harvest
Meat Raw, salted L. monocytogenes Texture, odour Slaughter
and smoked and spoilage lactic
ham & cooked acid bacteria
ham and paté
Fish Salmon fillets specific spoilage Texture Drum chilling of living
organisms salmon
Super chilled/super cooled
Fish Salmon fillets specific spoilage Texture Drum chilling of living
organisms salmon
Meat Pork neck L. monocytogenes Texture Slaughter
cutlet and spoilage lactic
acid bacteria
Frozen
Milk Ice cream - Texture, ice crystal Aging of pasteurized,
products size, sensory mixed ingredients
evaluation
Meat Pork meat - Texture, odour Slaughter
Vegetables Spinach - Vitamin C, texture, Freezing of spinach
colour, sensory pellets
evaluation

Heat transfer, energy usage and CO2 emissions along the reference cold chains
For each block of the reference cold chain, process values (such as inlet/outlet temperature, block duration, air
velocity) and cooling technologies involved are being listed. The best available technology (BAT) is chosen for
the different blocks of each reference cold chain. Data from the European cold chain will be used to link energy
usage and CO2 emission to the different technologies. Furthermore, a heat and mass transfer model for the room
air temperature needs to be coupled to the product heat and mass transfer model to enable prediction of product
temperature along the cold chain. The predicted product temperature is used as an input to kinetic models of the
different quality and safety indicators predicting quality and safety dynamics. The set point temperature and
process duration are used as inputs for BAT energy models to predict energy usage and refrigerant leakage
along the cold chain.

CONCLUSION
The framework will be used to specify the requirements of the user-friendly software, enabling to calculate the
effect of improving existing refrigeration technologies or incorporating emerging technologies in European cold
chain on the resulting energy usage, emissions and food quality and safety attributes.

ACKNOWLEDGEMENT: The research leading to these results has received funding from the European
Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 245288.

2246
 Influence of room temperature on food safety in refrigerated display cabinet

Laguerre O.a, Hoang M.a, Alvarez G.a, Flick D.b

a
Refrigeration Process Engineering, Cemagref, 92160 Antony, France
(onrawee.laguerre@cemagref.fr;hong-minh.hoang@cemagref.fr; graciela.alvarez@cemagref.fr)
b
AgroParisTech, 16 rue Claude Bernard, 75231 Paris Cedex 05, France (denis.flick@agroparistech.fr)

INTRODUCTION
A survey carried out by our team [1] showed that 30% of products presented in refrigerated
display cabinet were subjected to temperature abuse (more than 2°C higher than recommended
preservation temperature). Willocx et al [2] carried out a survey on processed vegetables in
Belgian retail display cabinets. This study also showed that retail display cabinets are a critical
point in the cold chain. Evans et al [3] observed that in open front display cabinet, the majority
of high temperature packs (97%) were located at the front and the largest number (60%) of
them was at the front base.
This work was carried out to; firstly, experimentally study the influence of the room
temperature on the product temperature in an open refrigerated display cabinet. Then, these
product temperatures were used in a predictive microbiological model to estimate the growth
of Listeria monocytogenes.

MATERIALS & METHODS

Figure 1 shows the side view of the display cabinet used in our study which was equipped with
one air curtain and 5 shelves. It was loaded with packages of test product made of
methylcellulose. Some packages were instrumented by calibrated thermocouples (T-type).
The display cabinet was located in a test room in which the room temperature was controlled at
20, 25 and 30°C. The temperature of air and test packages was measured every minute until the
steady state was reached.

RESULTS & DISCUSSION

The average temperature was calculated over 3h of the steady state period and reported in
figure1. The rise of room temperature leads to increase the air and the load temperatures
particularly at the front of the display cabinet.
A simple predictive model was used assuming a first order growth rate. The growth of Listeria
monocytogenes was estimated at various product temperatures after 4 days storage (Table 1).

CONCLUSION
Higher room temperature leads to higher air and product temperatures particularly the one
located at the front. This can be explained by the external air infiltration and the heat loss from
the display cabinet. The product temperature was used in a predictive microbiological model to
estimate the Listeria monocytogenes growth. This approach can be used as a tool of risk
evaluation.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2247
 a-Troom = 20°C b-Troom = 25°C c-Troom = 30°C
Figure 1. Product and air temperatures in the studied display cabinet.
bold and italic=air temperature, underlined= surface temperature of package, not underlined = centre
temperature of package

Table 1. Influence of product temperature on the growth of Listeria monocytogenes after 4 days storage.
Temperature Log[N(t)/N0)]
1.3°C (lowest observed product temperature) 0.13
4.0°C (maximal recommended storage temperature) 1.2
7.0°C 3.8
9.9°C (highest observed product temperature) 7.4

REFERENCES
[1] Cemagref & ANIA 2004. La chaine du froid du fabricant au consommateur: résultats de l'audit
ANIA/Cemagref. Revue Générale du Froid, 1042, 29-36.
[2] Willocx, F., Hendrick, M., Tobback, P., 1994. A preliminary survey into the temperature conditions
and residence time distribution of minimally processed MAP vegetables in Belgian retail display
cabinets. International Journal of Refrigeration, 17(7), 436-444.
[3] Evans, J.A., Scarcelli S. & Swain M.V.L. 2007. Temperature and energy performance of
refrigerated retail display and commercial catering cabinets under test conditions.
International Journal of Refrigeration, 30, 398-408.

ACKNOWLEDGEMENT
The research leading to these results has received funding from the European Community’s
Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 245288.

2248
 Improvement of existing concepts and refrigeration technologies: advanced control and

thermal energy storage applied to food Refrigeration

Denis Leducqa, P. Shalbarta, F. Trinqueta, A. Gracielaa, B. Verlindenb, S. van der Sluisc, E. Wessinkd, J.
EvanseTim Browne, jBart Nicolaïf, Annemie Geeraerdf. P. Verbovenf, J. M Lagarong, F. Jayh, M. Piranii, ,
E. Indergårdj

a
CEMAGREF, Antony, France (denis.leducq@cemagref.fr),b VCBT, Leuven, Belgium c SAINT TROFEE,
Renesse, The Netherlands d TNO, , Apeldoorn, The Netherlands, eLSBU, Langford, UK, f BIOSYST-
MeBioS, K.U.Leuven, Leuven, Belgium , g CSIC, Spain, h CRISTOPIA Energy Systems, France i SPES,
Fabriano, Italy, j SINTEF Energy Research, Trondheim, Norway

INTRODUCTION
In a context of greenhouse gas emissions, oil price rising and intermittent renewable energy
sources, energy storage, and more specifically thermal energy storage is one of the best
candidates to reduce and optimize the energy use of refrigerating systems. Moreover, the
temperature stability and the autonomy of those systems in case of power failure, related to the
use of thermal energy storage devices, is also an important factor of food quality and security
enhancement. The thermal energy storage (TES) technology has already attracted a number of
applications. From short-term storage in food containers to long-term storage in low
temperature warehouses, food engineering should also take a full advantage of its potential.
Coupled with control strategies as predictive control approach, it can lead to a drastic reduction
of energy consumption and a significant product quality enhancement. Through two cases, this
paper proposes to show the potential of those technologies for food applications.

MATERIALS & METHODS

Household refrigerator with thermal energy storage
The original experimental device is a single-compartment refrigerator. A phase change material
(PCM) slab is located on the back side of the evaporator. Temperatures were measured at
various locations on the refrigerating system, in the TES device and the cabinet. The
experiments have been realized inside an environmental chamber with temperature and
humidity controlled within 0.1°C and 1% fluctuations respectively. The value of the overall
heat transfer coefficient for the refrigerator is 0.44 W/m².K.
Dairy chiller with thermal energy storage capacity and advanced control
The second experimental device is an industrial dairy chiller with ice and chilled water storage
capacities. The refrigerating plant is split into three distinct systems. Two of them are dedicated
to water cooling and one to ice making. An advanced controller reads any measurement on
every programmable controller connected to the systems, and analyse the behaviour of the
global plant. Its action is based on modifying set points or directly controlling the main
actuators (for example, compressors), depending on the process.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2249
RESULTS & DISCUSSION
Household refrigerator with thermal energy storage
To evaluate the energy performance and the cool storage capacity of the refrigerator with and
without latent heat storage, the experiments have been performed for various thermal loads,
using no PCM, and water or an eutectic mixture as PCMs. For some specific runs, an
additional thermal load has been imposed using electric heaters, equipped with rheostat control
units for heat adjustment.
A first result observed was that the cycling period has changed from 180 mn without PCM to
540 mn with only a 5 mm PCM. This is a significant enhancement of the security for the
product since the integration of latent heat storage allows 5–9 h of continuous operation
without electrical supply (to be compared to 1–3 h without PCM). Another consequence is the
damping role for the food product temperature which has already been investigated in
literature, resulting in a food product quality enhancement.
A second significant result was a 10–30% increase of the coefficient of performance,
depending on the thermal load, mainly due to a higher evaporation temperature and thus a
higher cooling capacity.

Dairy chiller with thermal energy storage capacity and advanced control
Model predictive control (MPC) will be developed in FRISBEE to predict the future response
of the system taking into account scenarios such as energy availability and price, weather
forecast, provisional load of the production line, and the thermal inertia caused by the phase
change material.
A previous experiment on a dairy chiller has already shown the potential of the predictive
control approach. It consisted to implement a predictive controller using a physical model of
the plant, a criteria based on energy consumption and refrigerating demand, and finally a
possibility for the controller to modify remotely the set-points of the plant. After activation of
the controller, a 8% enhancement of the energy consumption has been observed, mainly due to
a much more adequate use of the three distinct systems.
In the last example, there was no model predicting the behaviour of the PCM, no foreseeable
events taken into account and the criteria, restricted to the energy consumption, did not take
into account any product impact. All these limitations will be overtaken in FRISBEE project.

CONCLUSION
By using a predictive control approach or using phase change material in refrigeration systems,
previous experiments have already shown significant enhancements on energy consumption.
By anticipating on the refrigeration demand, taking into account foreseeable scenarios, using a
criteria including quality of the product and energy performance, implementing a predictive
control approach and thermal energy storage devices, the FRISBEE project should allow to
develop an innovative and safer approach of controlling the refrigerating systems involved in
the cold chain application.

ACKNOWLEDGEMENT
The research leading to these results has received funding from the European Community‘s
Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 245288.

2250
Emerging Refrigeration Technologies at Laboratory Scale To Improve Food Quality And

Reduce Environmental Impact And Energy Consumption

Judith Evansa, Tim Browna, Denis Leducqb, Graciela Alvarezbb, Pieter Verbovenc, Bart Nicolaïc,
Annemie Geeraerdc, Edo Wessinkd, Ingrid Claussene, Erlend Indergårde, José Maria Lagarónf,h, Rocio
Pérez Masiáf,h, Stephane Moussetg, Alper Soysale, Marie-Christine Zelemj and Neil Wilsonk.

a
LSBU, Langford, UK (j.a.evans@lsbu.ac.uk)
b
CEMAGREF, Antony, France (graciela.alvarez@cemagref.fr)
c
BIOSYST-MeBioS, K.U.Leuven, Leuven, Belgium (annemie.geeraerd@biw.kuleuven.be)
d
TNO, Apeldoorn, The Netherlands (edo.wissink@tno.nl)
e
SINTEF Energy Research, Trondheim, Norway (Ingrid.C.Claussen@sintef.no)
f
CSIC, Burjassot, Spain (lagaron@iata.csic.es)
g
Costan S.p.A, Limana, Italy (stephane.mousset@eptarefrigeration.com)
h
NanoBioMatters, Burjassot, Spain (lagaron@nanobiomatters.com)
e
Arcelik, Istanbul, Turkey (alper.soysal@arcelik.com)
j
CNRS, Toulouse, France (zelem@univ-tlse2.fr)
k
Camfridge, Cambridge, UK (nwilson@camfridge.com)

INTRODUCTION
The Frisbee project (Food Refrigeration Innovations for Safety, consumers’ Benefit, Environmental
impact and Energy optimisation along the cold chain in Europe) will develop new tools and technologies
for use throughout the food cold chain. In work package 5 the Frisbee team will develop new and
emerging refrigeration technologies for representative cold chains selected for application in the
European food industry.

TECHNOLOGIES BEING CONSIDERED

There are a huge number of technologies that are being developed that may have applicability in the
refrigeration of foods. An evaluation of the available technologies that had applicability in the next 5-7
years resulted in the technologies listed in Table 1 being selected for development. Most of the
technologies are more suited to a certain sector of the cold chain and will be applied to the selected food
types being considered within the project (pork, salmon, apples, spinach or ice cream).

Table 1. Technologies being investigated in work package 5.

Sector Technology Sector of cold chain Food
Food Superchilling Primary chilling Pork
based Supercooling Primary chilling Pork
Smart packaging Retail/domestic All
Process Magnetic refrigeration Domestic All
based Air cycle refrigeration Blast freezing Pork, salmon, spinach,
ice cream
Nanoparticle refrigeration All Non specific
VIPs (Vacuum Insulated Panels) All Non specific

11th International Congress on Engineering and Food - Athens, Greece, 2011 2251
A short overview of each technology is contained below.
Superchilling and supercooling
Superchilling and supercooling have great potential to enable safe, high quality and long term storage of
foods without the consumer perceived detrimental effects of freezing. Superchilling allows 10-15% of the
free water in a product to be frozen whereas supercoooling enables all water to remain unfrozen. These
technologies are being considered by LSBU and SINTEF and will be combined with perfusion chilling
for meat where the additional benefits of rapid cooling, low weight loss and novel products are envisaged.
Smart packaging and VIPs
Work package 5 will develop 2 novel packaging technologies. LSBU will investigate VIPs for enhanced
thermal insulation for refrigeration systems and CSIC, Nanobiomatters and Cemagref will work on
nanoencapsulated PCMs (phase change materials) for food packaging.
VIPs have conductivities 5 times less than standard polyurethane insulation. However, their application
needs skill and the costs of panels still restricts uptake. Within work package 5 LSBU will develop
models and investigate reducing manufacturing costs. The work to develop PCMs within food packaging
will use composite nano-structured PCMs incorporated into packaging to provide thermal storage
capacity and to prevent unwanted temperature abuse of perishable food.
Magnetic refrigeration
Magnetic refrigeration exploits the magnetocaloric effect (the temperature change observed when certain
materials are exposed to a rapidly changing magnetic field) found in for example
gadolinium, lanthanum or manganese alloys. The real challenge in magnetic refrigeration is to increase
the temperature span of the refrigeration cycle. A key innovation has been the creation of a regenerative
cooling cycle, which extends the span of a magnetic refrigerator. In work package 5, Camfridge are
developing magnetic refrigeration for domestic and commercial refrigerators with the help of Arcelik and
Costan.
Air cycle refrigeration
LSBU are working to develop air cycle refrigeration for rapid freezing applications. Using air as the
working refrigerant has considerable potential or low temperature freezing applications. Air is a benign
working fluid and does not harm the environment or has the safety implications that are associated with
other refrigerants. There is considerable potential to develop air cycle systems based on optimised and
balanced components that would be suitable for demonstration in the food industry. The main areas
investigated will be fast freezing potentially combined with heating of hot water or food cooking.
Nanoparticle refrigeration optimisation
Within the project Cemagref will work on developing nanofluids for refrigeration system optimisation.
Nanofluids are engineered colloidal suspensions of nanoparticles (1-100 nm) in a base fluid that are used
to enhance heat transfer in conventional refrigeration. Large increases in heat transfer coefficients have
been observed by using only a low concentration of highly conductivity particles (carbon nanotubes).

CONCLUSIONS
Technologies within work package 5 will be used together with technologies from earlier work packages
as part of the demonstration and dissemination activities where optimised cold chains for pork, salmon,
apples, spinach and ice cream will be promoted.

ACKNOWLEDGEMENT: The research leading to these results has received funding from the
European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n°
245288.

2252
The potential for Superchilling to enable safe, high quality and long term storage of foods

I. C. Claussen

SINTEF Energy Research, Kolbjørn Hejes vei 1D, NO-7465 Trondheim, Norway
(ingrid.c.claussen@sintef.no)

INTRODUCTION
Superchilling is a concept where the temperature is reduced 1-2 °C below the initial freezing
point of the product. This results in a so-called ‘shell freezing’, where a thin layer of ice is
produced on the product surface during processing. The small amount of ice formed within the
product serves as a heat sink, eliminating the need for ice during storage and transport. As an
illustration, chilled haddock fillets have approximately 30 % higher environmental impact
potential than superchilled fillets due to the need for ice during storage and transport [1].
During storage, the ice distribution equalizes and the product obtains a uniform temperature
and the product appears as fresh. Consumer market analysis gives superchilled products as
good as or better quality score compared with chilled products.
The superchilling concept is not a new invention, and was described as early as 1920 by Le
Danois. Later, in the 1970’s and 1980’s, superchilling was mainly studied for transportation of
fish at sea. For the last 10-20 years the concept has been under continuous development.
Superchilling can be performed by means of several methods, RSW chilling (refrigerated sea
water), air chilling in blast tunnels and contact chilling being among the most used. The
Norwegian food industry is currently taking on the superchilling concept. In the meat industry,
superchilling is used ‘in-house’ in the industrial plant to expand the shelf life of the product,
ease the production and storage planning and to extend the sales period for fresh meat. In the
fish industry, superchilling of fillets increases the product yield and quality, resulting in more
of the raw material being sold as fresh fillets rather than frozen. For both industries,
superchilled conditions are applied only for the processing line and initial storage and the
advantages related to prolonged shelf life is to this day not fully exploited.
The objective of this work is to point out the main advantages and potential for the
superchilling concept to enable safe, high quality and long term storage of foods.

MATERIALS & METHODS

Several superchilling experiments have been performed on different food products to
demonstrate the extended shelf life of superchilled products. The extended sheld life consider
both the microbiological and physical changes during superchilling and storage of superchilled
products. A calorimetric method for measuring ice fraction has been established, and an online
near-infrared spectroscopy (NIR) method for measuring ice fraction is validated for prediction
of superchilled salmon [2]. The microbiological growth, by means of CFU (colony forming
units) is measured by method NMKL 96 [3]. Other physical changes such as drip loss, water
holding capacity and texture are measured by means of standardised methods in order to verify
quality changes in superchilled food products compared to chilled products.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2253
RESULTS & DISCUSSION
The shelf life of a food product is mainly defined based on a quality limit of 107 CFU/g, above
which food is regarded as unfit for human consumption. Figure 1 is based on several
experiments on superchilled and chilled food products and summarises the average shelf life
for the specific products due to the quality limit of 107 CFU/g.

Figure 1. Differences in shelf life of superchilled and chilled chicken, salmon and cod.

Figure 1 reveals that the shelf life of superchilled chicken and salmon is 50 % longer compared
to chilled product, while superchilled cod obtain 46 % longer shelf life compared to chilled
product. Since previous and ongoing research have already shown that superchilling of food
products does increase the shelf life and give high quality end-products, the main challenge
today is an efficient implementation and optimization of the superchilling processing lines in
the different food industries.

CONCLUSION
Superchilling enable safe, high quality and long term storage of foods. The main advantage is
the approximate doubling of shelf life for superchilled products compared to chilled products,
high product quality, higher yield and the potential for reduced environmental impacts
(approximately 30 %) when changing from chilled to superchilled value chains.

ACKNOWLEDGEMENT: The research leading to these results has received funding from the
European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n°
245288.
REFERENCES
[1] Claussen I.C., Indergård, E. & Grinde M. 2011. Comparative life cucle assessment (LCA) of production and
transport of chilled versus superchilled Haddock fillets from Norway to France. International Congress on Engineering
and Food, Athens, Greece, 22-26 May, 2011. To be published. [2] Stevik A.M., Duun, A.S., Rustad, T., O’Farrell M.,
Schulerud, H. & Ottestad S. 201. Ice fraction assessment by near-infrared spectroscopy enhancing automated
superchilling process lines. Journal of Food Engineering, 100(2010), 169-177. [3] Nordic Committee on Food
Analysis. 2003. NMKL Method 96. Bacterial Examinations in Fresh and Frozen Seafoods, third ed. [4] Stevik A.M., &
Claussen I.C. 2011. Industrial superchilling. A practical approach. International Congress on Engineering and Food,
Athens, Greece, 22-26 May, 2011. To be published.

2254
 CAFÉ : Computer-Aided Food processes for control Engineering

Dochain Denisa, Alonso Antoniob

a
CESAME, Université catholique de Louvain, Louvain-la-Neuve, Belgium
(denis.dochain@uclouvain.be)
b
CSIC, Vigo, Spain (antonio@iim.csic.es)

INTRODUCTION
The food industry is well established and many processes in operation nowadays are the
subject of intensive work with regard to the ways of devising better operation modes in terms
of product quality and safety (how to operate in order to ensure quality and comply with
safety constraints) as well as in terms of operation costs and environmental impact. There is
also an intensive development work aimed at responding to consumer demands by designing
new products and designing and operating the more appropriate combination of unit
operations needed to produce them.
However and despite the fact that the essential physical, biochemical and microbiological
principles are reasonably well understood, foods are complex systems with properties that
because are connected with quality and safety are usually very difficult to measure, estimate
or even represent through reliable models. Such properties may include physico-chemical
parameters associated to quality such as nutrient content, texture, colour or rheology, or
microbiological characteristics usually connected with food safety.
In addition, and from a Process Engineering perspective, the food industry integrates a rich
variety of apparently very diverse processes and technologies thus hampering the search for
unifying paradigms useful for dealing with different yet analogous processes. Such processes
have only recently been classified into a reasonably small number of categories, namely
bioconversion, separation, preservation and structuring.
The objective of the CAFÉ project is to provide new paradigms for the smart control of food
processes, on the basis of four typical processes in the areas of bioconversion, separation,
preservation and structuring. The novelty of the project lies in the capacity of combining PAT
(Process Analytical Technology) and sensing devices with models and simulation
environment with the following objectives:
1) to extract as much as possible information from the process/plant in the form of
precise estimations of unmeasured variables defining, in particular, product quality,
and of physical parameters changing as the process dynamics does or difficult to
know beforehand;
2) to save and encode in a reliable and usable way, basically via physical/deterministic
models;
3) to develop control methods to keep uniform quality and production despite the
variability in the raw material and/or to respond to sudden changes in the demand.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2255
 
The integration concept of CAFÉ

The four selected case studies are: wine making (bioconversion), microfiltration of food
beverages (separation), freeze-drying of lactic acid bacteria (preservation), and ice cream
crystallization (structuring).

The notion of paradigm is central in the CAFÉ project and serves as an integrating guideline
for most of the research activities within the project.

The consortium consists of sixteen organisations from seven member states. Due to the
ambitious technical nature of the project, five members of the consortium are universities
(UCL, APT, UTOV, WUR, UNIMAN) and three are research establishments (INRA, CSIC,
Cemagref), bringing to the consortium complementary skills, internationally recognised
expertise and a wealth of experience in European Union funded projects. The research
establishments also provide the link between academia and industry. One company (SPES)
gives the support for the development of the hardware sensors and for the hardware
integration of the project results. One company (Telstar) involved in the production of freeze-
dryers is participating in the implementation and evaluation of the tools developed in CAFÉ.
One company (PMS) involved in the manufacturing in freeze-drying products provides the
necessary expertise and facilities as end-users for the evaluation of the tools developed in the
framework of the present project. Three companies (C-Tech, Alctra, Norit) involved in sensor
development and commercialisation complement the sensor development activity and further
contribute to the integration and demonstration activities of the project. One company (BIV
Trace) provides expertise in traceability and quality management. And one company
(PSUTec) provides the necessary support for project management.

REFERENCES
http://www.cafe-project.org/

2256
 Design and development of REAlistic food Models with well-characterised micro- and

macro-structure and composition: DREAM

Monique Axelos

Head of the Science and Engineering of Agricultural Products division

Institut National de la Recherche Agronomique (INRA)

INRA Nantes, France (monique.axelos@nantes.inra.fr)

DREAM is aimed at developing realistic, physical and mathematical food models with well-
characterised structures, for use as standards to be exploited across all major food categories to
facilitate development of common approaches to risk/benefit assessment and nutritional quality
in food research and industry. These models will enhance the knowledge on process-structure-
property relationships from molecular to macroscopic levels. They will favour the creation of
generic food matrices, based on tailored microstructure to assess functional and nutritional
properties.
In terms of composition and structure, foods are very complex systems. Although scientists
have a good grasp of the former, the control of food structure remains difficult. But
understanding food structure is the key to understanding the effect of food on the human
health. There is an urgent need to improve our current knowledge on: i) the relationships
among food composition, processing, end-product structure and resulting material properties;
ii) the impact of these environment changes on nutriments and toxicants bioavailability as well
as on the microbial foodborne population and conversely, iii) the effect of the food microbiota
on the various food matrices.
The development of standard food models representing each a major food categories will make
it easier for public and private research partners to pool their knowledge and to enable partners
of the food industry, especially SME's, to benefit from models that are both generic and
realistic enough to optimize their existing processes or to come up with new ones. Scientists
also need generic but as realistic as possible models that can mimic food structure complexity.
Such models would make it much easier to assess the impact of a change in composition or of
processing conditions on the nutritional and health properties of foods and to help for
quantitative risk assessment studies.

To address the broadest possible range of food products and take into account their high
variability, foods will be classified in four generic structure groups:
· Filled cellular Solid (fruit and vegetables)
· Proteinous cellular network (meat);
· Combined gelled/dispersed/aerated systems (dairy products such as yogurts, creams and
cheeses)
· Open solid foam (cereal products such as bread).

11th International Congress on Engineering and Food - Athens, Greece, 2011 2257
Within these groups, the most relevant types of food products will be chosen using criteria
encompassing structural characteristics, industrial needs, and societal demands. In this way,
risks/benefits (food safety, nutritional aspects), economic weight and sustainability will be
taken into consideration. The fact that these four groups are based on structure rather than
specific products (open solid foam rather than bread for instance) will facilitate dissemination
and promotion activities through specific industry-sector associations, and encourage the
extension of the models to other food commodities.

The innovation brought by DREAM consists in applying cognitive science to integrate know-
how into scientific knowledge to the development of model foods and their standard operating
procedures (SOPs) for use by food industries, an up until now unprecedented approach.
Industrial partners (SOREDAB and UB) and five industry-oriented organizations (ADRIA,
CCFRA, CCHU, ACTILAIT and TIFN) are integrated into the project, contributing to
specification, providing validation feedback for overall improvement and standardisation.
The applicability of the model foods and food models will be assessed before transferring the
protocols and disseminating the gained knowledge to industry and other stakeholders
(EFFoST, The European "Food for Life" platform and national platforms, the CIAA and
national federations, EFSA and national regulatory bodies).

For each of these categories, three types of models will be developed:

_ Generic Model Foods (GMFs) are realistic physical models in which several parameters can
be varied, leading to a series of well defined samples for each given type of foods; GMF
fabrication protocols will be established; GMFs’ structure and chemical composition will be
determined and relationships between structure and chemical composition and functional
properties will be characterised.
_ Basic Knowledge Models (BKMs) are elementary food models describing specific aspects
of GMFs, through heuristic or mathematical approaches; for example, BKMs describe the role
played by temperature, pressure, chemical composition, etc. in a GMF’s structure and resulting
material properties.
_ Integrated Knowledge Models (IKMs) are dynamic networks - software systems –
integrating the operating rules of BKMs, technical expert knowledge, food properties and food
processing data from the GMFs. Results from initial experiments and simulations will be used
to improve IKMs’ mathematical models to reveal key parameters and material behaviour and
help refine GMFs: this iterative approach will optimise the food model concept prior to the
pilot stage.

Project website1 address: http://dream.aaeuropae.org/

1
The home page of the website should contain the generic European flag and the FP7 logo which are
available in electronicz format at the Europa website (logo of the European flag:
http://europa.eu/abc/symbols/emblem/index_en.htm logo of the 7th
FP: http://ec.europa.eu/research/fp7/index_en.cfm?pg=logos). The area of activity of the project should
also be mentioned.

2258
 Innovative technologies from Animal-by Products bioconversion

European project PROSPARE

Prof. Arnaldo Dossena a, Prof. Vladimir Popovb

a
Depts.Organic and Food Chemistry University of Parma, Parma, Italy (arnaldo.dossena@unipr.it)
b
A.N. Bakh Institute of Biochemistry of Russian Academy of Sciences, Moscow, Russia
(vpopov@inbi.ras.ru)

INTRODUCTION
PROSPARE -PROgress in Saving Proteins and Recovery of Energy (www.prospare.eu) is
a joint European-Russian research project, financed under the 7th Framework Programme (N.
212696, 2008 - 2011) for a total budget of 3.7M€ and an EU contribution of 2.7M€, committed
to the recovery of poultry industry leftovers into valuable end products. The PROSPARE
consortium includes eight research groups: four from the EU (two from Italy and two from
Belgium) and four from the Russian Federation. The consortium is coordinated by the
University of Parma (Italy), Department of Organic and Industrial Chemistry. The Russian
partners are coordinated by Bach Institute of Biochemistry of the Russian Academy of Sciences
(INBI).
The project worked at converting a problem into an added value bio-opportunity. In the
production of meat for human consumption, up to 50% of the animal weight is discharged, as
leftover [1]. This enormous mass from the meat industry has still raw materials rich in proteins
and lipids. Although this potentiality, most of this material is incinerated - only 22% is
converted into feed and barely 3% can become food [2].
The traditional rendering technologies, based on prolonged heating of the leftovers, ensure
microbial safety and increase digestibility, but use enormous amounts of energy and cause the
degradation of components of high biological value, at the same time inducing the formation of
compounds with undesired sensory properties and potentially harmful. Therefore, PROSPARE
project is aimed at development of complex technological platform for conversion of poultry
processing by-products into value added protein hydrolysates and biodiesel.

MATERIALS & METHODS

The technologies for processing the raw materials (feathers, bones, carcasses etc.) have been
developed by the Russian State Research Institute in Poultry Processing Industry (VNIIP) and
Symbol Ltd.. Technology of conversion of poultry meat &bone residues into functional animal
protein (FAP) is based on enzymatic hydrolysis of raw materials with enzyme blend under mild
pH (7.0) and temperature (about 55°C) conditions. The technology was optimized by
multifactor methodology with soluble protein recovery as key parameter monitored.
The Flemish Institute for Technological Research (VITO) developed a suitable procedure for
the transformation of lipids in biodiesel. Fat and tallow obtained after the hydrolysis were
subjected for biodiesel production by compact catalytic HTPM-process. The quality and
composition of fat fraction were compared to that of rendered chicken fat and rapeseed oil.

11th International Congress on Engineering and Food - Athens, Greece, 2011 2259
Optimal conditions of biodiesel production (temperature, duration, specific surface of catalyst)
were defined from multifactor experiment.
Unique high temperature short term (HTST) feather processing technology into feed raw
material – functional feather protein (FFP) was elaborated. The conditions of HTST treatment
were optimized aiming at maximal digestibility of FFP. In order to improve the solubility and
digestibility of FFP it was further subjected for enzymatic treatment. A.N. Bakh institute of
Biochemistry (INBI) and the group at the Department of Organic and Industrial Chemistry of
University of Parma have collaborated in the characterization of molecular composition (amino
acid composition, peptide profile, composition of volatile fraction) and functional (antioxidant,
antihypertensive, bifidogenic, antimicrobial) [3, 4] properties of the products and ensured their
safety concerns.
The integrated approach of the project was completed by the two partners as potential end-
users of the new technology: the Russian Mobitek-M, which develops food products from
protein hydrolysates, and the Italian Agricola Tre Valli, interested in producing high
digestibility feather hydrolyzates, for the pet-food industry. The consortium is completed by
CORE Biotech, a Belgian SME company specialized in functional proteins, contributing in the
dissemination and demonstration of the project results to potential industry sectors, for the
market exploitation of the new technologies across the EU and beyond.

RESULTS & DISCUSSION

The FAP- Functional Animal Protein multi-phase process developed under PROSPARE
Project, increases the peptides production to 85% high-quality peptide output from a poultry
post-slaughtering process; +42% of the current technology. The optimized technological
protocol developed provides >70% recovery of potentially available protein from poultry
leftovers Mild conditions of enzymatic hydrolysis ensure high retention of labile biologically
active compounds (e.g. thermo labile amino and fatty acids). The new FAP specific equipment
(follow up of the pilot system) can be installed as post-processor to the slaughtering production
line, with overall hi-profitability assured by hi-capacity and quality of the process and its
automation.
FAP is mainly composed of peptides and free amino acids (30%). FAP was characterised by
valuable amino acid composition with high content of lysine and tryptophan. Some, non-
proteolytic low molecular weight nitrogen-containing compounds that are characteristic for
meat products (anserine, carnosine, guanine, guanosine, adenosine, creatine, creatinine) were
also identified in FAP. Volatile fraction of FAP, that is essential for sensory properties, is
composed of fatty acids and different diketopiperasines. FAP exhibits a wide spectrum of
biological effects, including antioxidant (TEAC and ORAC values around 650 and 320 Pmol
TE/g) and antihypertensive (IC50- ) and growth stimulating activity with respect to
lactobacilli.
FAP was characterised with low fat content (<4%), low osmosis, high solubility under wide pH
range (300 g/L), complete digestibility in vitro, and hypoallergenic properties, and could be
used for different application: dietetic, nutritional, extra supply, cosmetics.
Related to the FFP-Functional Feather Protein technology, a new patented hydrothermal
treatment of the feathers, has been developed which produces, during the first stage of the
process (<90 sec. of treatment, instead of hours), FFP from keratin. FFP have shown to be
tasteless, high premium feather’s meal (>85% digestibility, <2% fat and ash). The conditions

2260
Optimal conditions of biodiesel production (temperature, duration, specific surface of catalyst)
were defined from multifactor experiment.
Unique high temperature short term (HTST) feather processing technology into feed raw
material – functional feather protein (FFP) was elaborated. The conditions of HTST treatment
were optimized aiming at maximal digestibility of FFP. In order to improve the solubility and
digestibility of FFP it was further subjected for enzymatic treatment. A.N. Bakh institute of
Biochemistry (INBI) and the group at the Department of Organic and Industrial Chemistry of
University of Parma have collaborated in the characterization of molecular composition (amino
acid composition, peptide profile, composition of volatile fraction) and functional (antioxidant,
antihypertensive, bifidogenic, antimicrobial) [3, 4] properties of the products and ensured their
safety concerns.
The integrated approach of the project was completed by the two partners as potential end-
users of the new technology: the Russian Mobitek-M, which develops food products from
protein hydrolysates, and the Italian Agricola Tre Valli, interested in producing high
digestibility feather hydrolyzates, for the pet-food industry. The consortium is completed by
CORE Biotech, a Belgian SME company specialized in functional proteins, contributing in the
dissemination and demonstration of the project results to potential industry sectors, for the
market exploitation of the new technologies across the EU and beyond.

RESULTS & DISCUSSION

The FAP- Functional Animal Protein multi-phase process developed under PROSPARE
Project, increases the peptides production to 85% high-quality peptide output from a poultry
post-slaughtering process; +42% of the current technology. The optimized technological
protocol developed provides >70% recovery of potentially available protein from poultry
leftovers Mild conditions of enzymatic hydrolysis ensure high retention of labile biologically
active compounds (e.g. thermo labile amino and fatty acids). The new FAP specific equipment
(follow up of the pilot system) can be installed as post-processor to the slaughtering production
line, with overall hi-profitability assured by hi-capacity and quality of the process and its
automation.
FAP is mainly composed of peptides and free amino acids (30%). FAP was characterised by
valuable amino acid composition with high content of lysine and tryptophan. Some, non-
proteolytic low molecular weight nitrogen-containing compounds that are characteristic for
meat products (anserine, carnosine, guanine, guanosine, adenosine, creatine, creatinine) were
also identified in FAP. Volatile fraction of FAP, that is essential for sensory properties, is
composed of fatty acids and different diketopiperasines. FAP exhibits a wide spectrum of
biological effects, including antioxidant (TEAC and ORAC values around 650 and 320 Pmol
TE/g) and antihypertensive (IC50- ) and growth stimulating activity with respect to
lactobacilli.
FAP was characterised with low fat content (<4%), low osmosis, high solubility under wide pH
range (300 g/L), complete digestibility in vitro, and hypoallergenic properties, and could be
used for different application: dietetic, nutritional, extra supply, cosmetics.
Related to the FFP-Functional Feather Protein technology, a new patented hydrothermal
treatment of the feathers, has been developed which produces, during the first stage of the
process (<90 sec. of treatment, instead of hours), FFP from keratin. FFP have shown to be
tasteless, high premium feather’s meal (>85% digestibility, <2% fat and ash). The conditions

11th International Congress on Engineering and Food - Athens, Greece, 2011 2261
REFERENCES
[1] Arvanitoyannis I.S. & Ladas, D. 2008. Meat waste treatment methods and potential uses.
International Journal of Food Science and Technology, 43, 543-559.
[2] Lui D.-C. Better utilization of by-products from meat industry ROC 2002-10-01.
[3] Wu H. C., Pan B. S., Chang C. L., Shiau C. Y. 2005, Low-molecular Weight Peptides as
related to antioxidant Properties of Chicken essence. J. of Food and Drug Analysis, 13, 176-
183
[4] Eu Project SEAFOODplus 6th FP

Optimization of FFP enzymatic hydrolysis conditions

2262
 The Animal by Product (AB-P): challenging problem and resource

W. De Roover

GePro member of AVEC

Approach and objectives of the PROSPARE project

O. Koroleva

Bakh Inst. Biochemistry, Russian Academy Science, Moscow

Innovative methodology and process technologies

O. Koroleva

Bakh Inst. Biochemistry, Russian Academy Science, Moscow

Molecular composition and Functional Properties of Poultry Hydrolyzates obtained in

the PROSPARE Project

Arnaldo Dossena

University of Parma, Italy

Food & feed market exploitation and nutrition

Virgilio Guardiani

International nutrition consultancy

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2264
 AUTHORSINDEX

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-1
AbKadir,M.O.  213,259,1697 Almeida,H.   1227
Abakarov,A. 1399,1403,1523,1525 Almeida,I.   1263
Abavi,A.   1885 Almeida,P.I.F.  327
Abbaszadeh,R.  1779 AlmeidaPorciuncula,B.D. 579
AbdElͲGader,S.  653 Almonacid,S.F. 529,541,1407,1521,
AbdalaGomide,C.  1179 1815,1973,1977
Abdellaoui,H.  415 AlͲMuhtaseb,A.H.  253
Abdellatief,A.  93 Alonso,A.A. 281,805,1409,1531,
AbdulMajid,A.M.S.  213,259,1697 1537
Abdullah,A.  153,1119 AlͲRawi,S.S.  213,259,1697
Abdullah,N.  387 Alsaed,A.K.   653
AboͲSalah,K.M.  1697 Alsaffar,A.A.  43
Abreu,M.   1647 AlͲSaidi,G.   153,1119
Aburatan,M.  1935 AlͲSanabani,A.  1089,1981
Acevedo,C.A.  1259 AlstromͲMoore,A.  467
Acevedo,F.   829,2185 Altan,A.   901,1155
Achaerandio,I.  651 AltenhofendaSilva,M.  1007,1017
Achir,N.   689 Alvarenga,N.B.  1181
Adachi,S.   927,1351 Alvarez,G. 55,523,535,595,
Adamiak,A.  1763 1531,1841,2241,
Adamopoulos,K.G.  571,1955,1957 2245,2247,2251
AdlerͲNissen,J.  269,517 Alvarez,I.   325,1529
Afshari,M.   1907 Álvarez,K.   2103
Agalioti,M.   2087 AlvarezQuintero,R.  2015
Aghili,S.R.   1245 AlvarezͲJubete,L.  1773
Agoulon,A.   1615 Alves,C.C.O.  1693
AguedaCastro,M.  1251 Alves,V.   2061
Agüero,M.V.  1709,1811 Alves,V.D.   103,1023
Aguilar,L.   1637 Alzamora,S.M. 1251,1311,1629,
Aguilera,J.M. 67,115,193,279,289,781, 1635,1795
921,929,935,985,1231, Amante,E.R.  1085,1839,2181
1753 Amendola,D.  165,377
Ahmad,R.   653 Ammar,J.B.  909
Ahmad,U.   547 Amorim,E.O.C.  1733
AhmadTarmizi,A.  655 AmorimRamos,V.  939
Ahmadi,H.   1779 Ananingsih ,V.K.  647
Ahrné,L.   783,1975 Anantheswaran,R.  105
Akcin,A.   1891 Andés,A.   445
AkemiMakiyama,P.  1527 Andoa,H.   1901
Akgun,A.   1891,1893 Andrade,P.   1861
AkŦllŦoŒlu,G.  1807 Andrade,R.D.  1325
Akpinar,O.   2051 AndradeͲMahecha,M.M. 1009
Akterian,S.   1599 André,S.   1181
Alakali,J.S.   239 Andrés,A.   1993,1995
AlamillaBeltrán,L.  1163,1287,1393 Andresen,T.  315,1539
Albano,K.M.  225 Andrianyta,H.  457
Albers,D.   2224 Andrieux,J.C.  243,1137
Albertsson,P.ͲÅ.  2047 AnelauskaitĦ,E.  1271
Albors,A.   41 Anese,M.   177
Albuquerque,C.L.C.  2113 Anestis,S.   1103,1923,2147
AlcaideMarzal,J.  661 Àngel,B.   1277
Alcañiz,M.   1483 Angelidis,A.S.  1797
Alcântara,L.A.P.  1705 Angelov,M.  1451
Aldoomy,H.  653 Ángulo,M.   2109
Alegria,C.   1647 Anguy,Y.   1463
Alex,R.   415,955 Anjum,F.M.  1117
Alexandrakis,Z.  1683 Antelo,L.T.   805
Alexandre,E.M.C.  1625 Antonio,A.   2255
Alexopoulos,A.  1013,2029 AntôniodeMoraisJrc,M. 2137
Ali,I.   1207 Antoniou,K.D.  231,1301,1307
Ali,M.F.   887 AparecidadeCarvalho,R. 37,861,1179
Aligiannis,N.  2215 AparecidaGuaraldoGonçalves,L. 1047,1049,1293
Alkuino,L.D.  1929 AparecidaMaieves,H.  1085
Allen,P.   237,1631 Aparicio,G.   959
Almada,C.A.  1195 Apostolidi,S.  1337

I-2
Araki,T.   455,1281 BakŦrcŦ,F.   851
Arango,P.   167 Baks,T.   725
Arapaki,S.   1831 Balaban,M.O.  425,1453,1455
Araujo,E.A.F.  1227,1233 Balabanova,T.  2159
Araus,K.   443 Balagué,C.E.  973
Aravena,R.   1717 Balasubramaniam(Bala),V.M. 345
ArayaͲFarias,M.  1723,1735 Balestra,F.   221
Arballo,J.R.   1441 Balian,S.C.   1181
Arellano,M.  55,523,1531 BalsaͲCanto,E.  281,1531
ArenasͲOcampo,M.L.  1287 Bambicha,A.R.  613
Argyropoulos,D.  819,955 Baquero,R.   1325
AriasͲMendez,A.  281 Baranda,A.   741
Arisseto,A.P.  1875,1877 Baranowski,Piotr  1759
ArjonaͲRomán,J.L.  417,1129,1221 Barao,C.   917
Arnault,I.   1223 Barat,J.M   181,1483
Arnold,M.   1417 BarbadeAlba,L.R.  1027
Arratia,C.   1075 Barbier,C.   1579
Arrieche,L.S.  1821 BarbosaNeto,A.M.  2003
Arshad,A.   1117 BarbosaͲCanovas,G.V.  801
Arshad,M.U.  1117 Barla,F.   2119
Arsianti Y.  29 Barón,P.J.   1395
Artíguez,M.L.  353,741 Barreiro,P. 245,845,1477,2231
ArzateͲVazquez,I.  1045,1753 Barreto,I.M.A.  1097
Arzeni,C.   407 Barrio,Y.   1655
Aschenbrenner,M.  149,957 Barrios,S.   1005
Ascheri,J.L.R.  2151 BarryͲRyana,C.  453
Aspé,E.   2101,2103 Barta,J.   2213
Assifaoui,A.  875 Bartolomeoli,I.  1731
Asteriadou,K.  711,1555 Baša,L.   1547,1849
Astruc,T.   1855 Basch,C.   979
AtaçMogol,B.  1581,1807 Basios,A.   1273
Atarés,L.   857,859 Bassinello,P.Z.  1421,2129,2151
Athes,V.   807 Bassirou,B.   85
Atoniuk,A.   843,2227 Basso,L.C.   2107
Atungulu,G.  795 Batista,E.A.C. 1037,1125,1389,
Atuonwu,J.C.  1607 1391
Aubourg,S.   1737 Batista,F.R.M.  1389,1391,1405
Auger,J.   1223 Batu,A.   2117
Augustin,W.  701,703,705 Baudrit,C.   323
Augusto,P.E.D.  1299,1911 Baysal,A.H.   219
AvalloneBueno,L.  2085,2137 Bazinet,L.   1723,1735
AvelinoPasa,A.  953 Beatty,E.   1943
Avérous,L.   111 Beaulieu,L.   1723
Axelos,M.   1547,2257 Beck,M.   885
Ay,N.   2225 Becker,T. 25,79,233,885,1473,
Ayadi,F.   597 1513
Ayalla,J.V.   1953 Bednáriková,A.  1881
AytaƔAkcin,T.  1891 Bedoui,I.D.   415
Azevedo,S.   971 Behsnilian,D.  21
Azuara,E.   1639,2097 BeiraghiToosi,S.  1497,1499
Babahmetoviđ,L.  867 BeirãoͲdaͲCosta,L.  873
Bacelos,M.S.  327 BeirãoͲdaͲCosta,M.L.  673,793
Bach,S.   109 BeirãoͲdaͲCosta,S.  673,793
BadanRibeiro,A.P. 1047,1049,1065,1293 Belagardi,M.  1709
Badeka,A.V.  1209 Belessi,C.ͲI.A.  1829,1831
Bader,S.   589 Beletsiotis,E.  495,1765
Bae,Y.   1145 Bellalta,P.   921
Baez,O.   1369 Bellmann,C.  707
Baeza,S.   1087 BelloͲPérez,L.A.  1315
Bai,J.ͲW.   2013 BelšēakͲCvitanoviđ,A.  865,867,2039
Baier,D.   355,357 BenNama,M.M.  213,259
Baier,M.   393,1643 Benavides,S.  113
Bakalis,S. 49,147,265,329,533,621, Benezech,T.  713,715,1557
817,887,913,915,1511, Bengtsson,H.  223
1543,1887 Benites,C.I.   2063

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-3
Benkhelifa,H.  55,523,1531 Braun,S.   765
Benning,R.   569 Brennan,C.   1651,2157
Bergenståhl,B.  587,1169,1305 Brianceau,S.  1735
Bergin,D.   507 Briassoulis,D.  849
Beristain,C.I.  1639,2097 BrionesͲLabarca,V.  1675
Bernardo,C.  1085 Broeze,J.   301
Bernardos,A.  181 Bronlund,J.E. 337,633,1153,1429
Bernáškova,J.  1165 Brooker,A.D.M.  709
Bernat,N.   217,1335 Brown,T.   2245,2249,2251
Berski,W.   1255 Broyart,B.   483,1867,1921
Besbes,E.   1205 Bruyninckx,W.  499
Betz,M.   675,1687 Bubník,Z.   1165,1695,1707
Bez,J.   589 Buckow,R. 335,469,1417,1535
Bezirtzoglou,E.  1013,2029 Bucur,A.   1095
Bhattacharya,S.  273 Budiastra,I.W.  457,477
Biau,N.   201 BuenodaSilva,J.  2107
BibbinsͲMartínez,M.D.  1587,2021 Buera,P.   151
Bicas,J.L.   1727 Buffa,M.   1467
Bilbao,A.   1783 Bugarski,B.   173
Biliaderis,C.G. 33,101,129,143,879,1055, Bui,V.A.   557
2207 Bult,J.H.F.   611
Bindrich,U.   569 Bulut,S.   1627
Bird,M.   603 Bund,R.   87,727,779,1213
Bittante,A.M.Q.B.  37,861,1179,1329 Bunya,M.   1721
Bitti,M.T.   1571 Burmester,K.  1989
Blahovec,J.   39,163 Bustamante,A.  1717
Blank,I.   845,2231 Bustamante,J.  541
Blel,W.   713,1557 Bustos,N.   1815
Bobe,U.   687,2211 Bustos,R.   1087
Bodini,R.B   883 Butler,F.   473,497,507,1819
Bogiceviđ,B.  825 Byrne,E.   427
BoguzsJr,S.  2179 Byrne,H.   1773
Bohuon,P.   481,689,1349 C.Jurkiewicz,C.  2093
Bolouri,B. 95,997,999,1497,1499 Cabral,F.   2071
Bolourian,S.  1907 Cabral,L.M.C. 551,1641,1967,2203
Bonazzi,C. 481,483,1805,1867 Cabral,R.A.F.  1347
Bongaers,E.  843,2227 Cadioli,M.G.B.  1375
Bonilla,J.   857,859 Calado,V.   1879
BonillaͲReyna,B.  837 CaldasFonseca,S.  971
Bonnier,F.   1773 Calderon,M.  1603
Bonomo,P.   1487 Calderón,J.   1777
Bonomo,R.C.F.  1487 CalderónDomínguez,G.  1045
Boom,R.   65,203,719,725 Califano,A.N.  137
Boons,K.   501 Calligaris,S.   177
BorgesLaurindo,J.  579,953 Callon,C.   1947
Bornhorst,G.M.  251,631 Calvimontes,A.  707
Boros,D.   2149 Camitan,N.O.  1929
Borralho,E.   1181 Campanella,O.H.  135,1321
Borrin,T.R.   19 Campañone,L.A.  1441
Boscarioli,M.P.M.  2093 Canada,J.S.B.  1181
Bouchon,P. 123,1073,1077,1079,1365 Candal,R.J.   923
Boudouvis,A.G.  97 Cano,C.J.C.   389
Bouhallab,S.  561,1149 CanoͲSarmiento,C.  1163
Boulekou,S.  359 Canut,A.   1563
Bouraia,M.   731 Cao,X.   1659
Bourbon,A.I.  293,673,1015 Cao,Y.   1985
Bourgaux,C.  119 Cappuyns,A.M  471,475,499,1789
Bourke,P.   409 Carareto,N.D.D.  1131,1133
Boushey,C.   187 Caravelli,A.H.  743
Boxler,C.   705 Carballo,S.   1305
Boz,Z.   1485 Cárcel,J.A.   1951,1997
Brambilla,A.  2075 Cardenas,F.  615
BrancoShinagawab,F.  2085 Cardoso,C.   827
Brand,J.   397 Cardoso,L.P.  19
Brandão,T.R.S.  479,1625 Cardoso,V.L.  1711

I-4
CardosoDeOliveira,D.  1085 Charalampopoulos,D.  679,1793
CardosoReis,R.  2031 Charbonnel,R.  721
CardosoTramonte,K.  2181 Charinpanitkul,T.  255,519,2171
Carignano,M.A.  135 Charlemagne,N.  85
Cariso,A.R.   1929 Charoenrein,S.  1063,1127
Carle,R.   617 Chassagne,D.  373
CarmelloͲGuerreiro,S.M  1067 ChassagneͲBerces,S. 245,843,845,1175,
Carmona,P.A.O.  1269 2227,2229,2231
Carneiro,H.C.F.  1387 Chatziantoniou,S.  1317,1865
Carola,T.   1181 Chatzikyriakidou,K.  1845
Carpenco,J.  979 Chaudhuri,S.  379
Carr,A.   185 Chaves,M.A. 1097,1397,1487,
Carrer,C.C.   1329 2031
Carretero,C.  1239,1241 ChávezMontes,E.  649
Carrion,F.P.  1143 Cheison,S.   2099
CarvajalͲMillán,E.  2011 Cheison,S.C.  397
CarvajalͲRondanelli,P.A.  615 Chen,S.H.   1913
Carvalho,A.V.  1421,2129,2151 CherrezMaríaJosé,P.  1197
Carvalho,J.M.P.  27 Chesterton,A.  835
Carvalho,M.  275 Chevallier,S.  199,1545
Carvalho,R.A.  883,1033,1143 Chew,J.   603
Carvalho,R.N.  2151 Chew,Y.M.J.  709,1559
Castelló,R.   1023 Chida,K.   2223
Casteran,C.  243,1137 ChihMing,C.  1065
Castillo,M.   317,1467,1469 Chinelate,G.C.B.  2155
CastilloHernandez,D.  1587 Chiralt,A. 41,217,857,859,871,
Castro,A.   1305 1335
Castro,D.   1777 Chiron,H.   133,1113
Castro,E.   1971 Chiumarelli,M.  853
CavalcanteBarros,F.F.  739 Cho,Y.ͲJ.   823,1459
CavalcanteFai,A.E.  401,2107 Choi,W.   435
CavalcanteMaciel,T.  657 Chombeau,M.  1455
Cavalcanti,R.N.  2167,2189 Chranioti,C.  1291,1775
Cavallieri,Â.L.F.  933 Christakopoulos,P.  2215
Cavella,S.   1211,2069 Ciccarone,R.  371
Cavicchioli,B.  739 Ciesarová,Z.  1591,1881,1965
Cayot,P.   875 Cieslak,B.   1889
Celis,D.   1159 Cifuentes,N.  1229
Cenard,S.   267 Cipriano,A.   1753
Cerecero,R.  1603 CŦrban,D.   1453,1455
Ceriani,R.   1419,1953 Citeau,M.   1215
Cernela,J.   1921 Clark,J.P.   3,543
Cerqueira,M.A.  1015 ClarodaSilva,R.  1047
CésarDacanal,G.  2095 Claussen,I.C. 511,1905,2245,2251,
Cevoli,C.   1423 2253
ChacanaͲOjeda,M.  1675 Cobos,A.   985
Chacko,J.   105 Cocan,I.   1909
Chafer,M.   217,1335 Cocci,E.   221
Chaiwanichsiri,S.  677,1613 Cocco,D.   2205
Challois,S.   125 Cocco,G.   2205
Challou,F.   533 Cocolin,L.   2211
Chambi,H.   1139,1179 Coelho,M.T.  1647,2061
Chammem,N.  2211 Coelhoso,I.M.  1023
Chamorro,M.C.  1477 Cohen,R.   117
Chang,K.S.   1461,1565,1567 Cole,P.   711
Chang,Y.H.   1461,1565,1567 Collignan,A.  689,1349
Changli,H.   2037 Comaposada,J.  1415
Chanier,L.   133,1113 Condori,J.L.  193
ChanonaPérez,J.J. 623,1753,1045,1999 Connelly,R.K.  727
Chantoiseau,E.  295,1493,1495 Connolly,J.   427
Chanvrier,H. 245,843,845,1175,2227, Contini,C.   1001
2229,2231 Cook,M.T.   679
ChaparroGonzalez,M.P. 1197 Coral,D.F   1135
Char,C.   1827 Cordeiro,C.   103
Charalambides,M.N.  189,191,1275 CordeiroCardoso,J.  2131

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-5
CordeiroMancini,M.  2001 Dagà,I.   1241
Córdova,A.   1479,1813 Dalay,O.   169
Cornacchia,L.  925 DallaRosa,M. 221,565,771,797,
Cornet,I.   499 1011,1423
CoronadoJorge,M.F.  37,861 Dalmolin,I.   1037,1125
Coronel,C.P.  1791 DanMironescu,I.  1377
Correa,E.C.   1477 Danao,M.ͲG.  317
CorreaMendonç,A.  2065 Danilov,N.P.  663
CorreaMoreiraRocha,D.G. 2001 DapēeviđHadnaĜev,T.  1109,2141
Corredig,M.  421 DaraeiGarmakhany,A.  1751
Correia,P.   873,1949 Datta,A.K. 291,305,1449,1401,
Corrieu,G.   1947 1507
Cortellin,G.  987,1577,1937 Datta,C.   379
Cortés,M.   1971 Daudin,J.D.  965,1855
Cortés,P.   1365 David,T.   1817
Costa,E.   1263 Davies,C.E.   51
Costa,M.C.   1131,1133,1701 DeA.Bezerra,R.R.  2155
Costa,N.   1023 DeAlmeida,A.R.F.  333,1489
Costa,P.A.   1329 DeAndrade,C.J.  1589
Costa,R. 769,771,773,1553,1861 DeAndradeMattietto,R. 1437
CostaLima,Rui  665 DeAraujoMantovani,R.  933
CostaSilva,M.  1553,1861 DeAssisPerrechil,F.  939
Cotillon,C.   2241 DeBaerdemaeker,J.  209,1057
Courel,M. 483,489,1805,1867 DeBonis,M.V.  1945
Courtois,F.   311 DeCarvalho,D.S.  2179
Coutinho,J.A.P.  1703 DeCesaro,A.M.  1037
Cox,P.   533 DeFátimaFonseca,M.  2085,2137
Cox,P.W.   1043 DeFaveri,D.M.  165
Craig,A.P.   1761 DeGregorio,C.  743
Crisostomo,J.  1087 DeJesúsPereaFlores,M. 1999
CristianaͲNunes,M.  873 DeJong,P.   723
Cristianini,M. 1299,1661,1663,1733, DelaFuente,J.C.  443,1363
1911 DelaFuente,L.  1289
Cristina,I.   37 DelaLlera,A.A.  193
Cristinapetry,F.  1161 DelaTorreR.Rene,R.  445,1619,2021
Crivellari,G.P.  1445 DeLamoͲCastellví,S.  2193
Croguennec,T.  1149 DeLandeta,M.C.  1195
CrostinaCorrea,E.  845,2231 DeMoura,S.C.S.R.  1799
CruzXimenes,G.  2137 DeMouraGuimarães,P.C. 2153
Cuadros,T.   1231 DeO.Rios,A.  1421,2129
Cude,L.   1201 DeOliveir,C.M.  1085
Cuellas,A.   1691 DeOliveira,A.L.  1671
Cuevas,M.S.  1701 DeOliveiraPinto,C.L.  1823
Cullen,P.J. 285,409,427,1465,1651 DeOliveiraRomeraa,C.  953
Cunha,L.M.  665,971 DePilli,T.   1933,179
Cunha,R.L.   1069,1071 DePinho,S.C.  747
Cunningham,D.  229 DeRoode,M.  723
Curet,S.   1433 DeRoover,W.  2263
CuretͲPloquin,S.  1545 DeSouza,L.L.  1669
CurveloPereira,N.  1235 DeSouzaCastro,P.  1527
Cutolo,R.S.   1747 DeSouzaFerreira,J.  1747
Cuvelier,G.   195,201 DeSouzaSantana,R.S.  747
Cuvelier,M.E.  47 DeVito,F.   889
Cybulska,J.   1053 DeVries,R.   11,1041
DaCosta,R.L.  1727 Debeaufort,F.  373
DaCruzFrancisco,J.  1169 Defraeye,T.   2233
DaMatta,V.M.  1437 Dehghannejad,N.  1297
DaPenhaPiccoloRamos,M. 1823 Dejmek,P.   59,515
DaPieve,S.   177,1731 Dekker,M.   505
DaRochaFerreira,E.H.  1879 DelCarmenAltamiranoFortoul,R. 199
DaSilva,A.B.  333,1489 DelToro,G.V.  623
DaSilva,J.N.  1397 DelValle,G.  133,1113,1427
DaSilva,M.A.  1747 DelValle,J.M. 443,1363,1715,1717,
DaSilvaArrieche,L.  275 1719
DabinaͲBicka,I.  563 Delgado,A.   521,569

I-6
Delshad,M.  1779 Dostálová,J.  1165
Delwiche,M.  795 Douania,I.   267
Demarchi,S.M.  2041 Doursat,C. 125,295,1435,1443,
DemertziͲAkrida,K.  1013 1493,1495
Demertzis,P.G.  1013,1301 Doval,V.C.   1733
DenBesten,H.M.W.  493 Dowling,D.P.  1001
Denis,N’dri   85 Downey,G.   911
Depypere,F.  2233 Doxastakis,G.  1243
Dermesonluoglu,E.  1853 Doyen,A.   1723
Dermiki,M.   377 Drakakis,K.   451
Derossi,A.   179,1933 Drosinos,E.H.  1829,1885
Dervisoglu,M.  1295,1891,1893 Druon,C.   415
DesMarchais,L.ͲP.  1261,2033 Drusch,S.   63
Descours,E.  1919 Du,Z.ͲL.   2013
Desjardins,Y.  1735 Duarte,C.   673
Despotoviđ,S.  2057 Duarte,C.M.M.  673
Dessev,T.   199 Dubey,B.N.   643
Destandau,E.  809 Dubois,C.   197
Devlieghere,F.  475 Ducept,F.   195
Dewettinck,K.  2233 Dueik,V.   1073
DeyaniraVegaMéndez,D. 1111 Dukalska,L.   1199
Dhall,A.   291,1401,1507 Dumoulin,E.  47,1463
Dhamole,P.B.  1699 Dupuy,A.   807
Dheeman,D.S.  2209 DuránPáramo,E.  623
DiScala,K.   2049 Dushkova,M.  1083
Dias,J.M.   1181 DustedMendoza,J.C.  1739
DiasDeCastanhoAmboni,R. 2181 Dutheil,E.   1651
Diaz,O.   985 Duthoit,M.   1647
Díaz,A.I.   1783 Dutta,A.   379
Díaz,J.   1777 Dutta,D.   379
Díaz,P.   1075 Duxenneuner,M.R.  643
DíazNarváez,G.C.  1027,1963 East,A.R.   1153,1873
DíazͲCarvajal,D.  2021 EbrahimzadehMousavi,S.M.A. 95,997,999
Dima,J.B.   1395 Echegaray,E.  1971
Dimitreli,G.  231,693,1307 Edris,A.E.   175
Dimitrijeviđ,S.  825 Efraim,P.   1799,2145
Dimou,A.   341 Efremov,G.   491
Dionísio,A.P.  1725,1727,2179 Eggers,R.   601,1715,1989
Diosady,L.   667,759 Egito,A.S.   1641
Djordjevic,R.  2143 Ehlbeck,J.   1643
DoAmaralSobral,P.J.  37,861,1009,1179 Eisner,P.   589,617
DoCarmoFerreira,M.  1327,1359 Ekiz,H.I.   585
DoPrado,M.M.  1367 Elfakir,C.   809
Dochain,D.   2255 Elfawakhry ,H.  233
Dodd,C.   467 ElitaLópez,M.  1289
Doehner,I.   355 ElMasry,G.   237,1631
Dogan,H.   1151 ElͲSamahy,S. 607,1089,1091,1979,
Dogenski,M.  23 1981
Dokiđ,L.   881 Emadzadeh,B.  1093
Domingues,C.  1949 EmamͲDjomeh,Z.  271,893
Domingues,R.  1711 EmanuelssonͲPatterson,E. 211
DongChen ,X.  297,1505 Emek,S.C.   2047
Donsì,F.   165,349,681 EncisoͲChavez,B.  1587
Donsì,G.   1657,1729 Enomoto,T.  1935,2119
Dorantes,L.  959 Enrione,J. 141,1075,1087,1159,
DorantesͲAlvareza,L.  961 1183
DorantesͲNieto,A.  2021 Erabit,N.   535,595
Dornier,M.   481 Erdogdu,S.B.  585
Dorofejev, K.  1199 ErdoŒdu,F.   331,1411,1485
DosReis,C.   941 Erkinbaev,C.  139,1057,2239
DosReis,R.C.  1725 ErlansonͲAlbertsson,C.  2047
DosSantos,R.  2107,2179 Escalona,V.  1827
DosSantos ,S.B.  1703 Eschenhagen,U.  707
DosSantosSilva,V.L.  735 EscobedoͲAvellaneda,Z.  1671,2139
Dossena,A.   2263 Escola,H.   1181

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-7
Eshraghi,E.   1497,1499 Ferrentino,G.  425
Esmaiili,M.   241,1167,1713 Ferret,E.   1919
Espigulé,E.   1239,1241 Ferrua,M.J.   513,619,629
Espinosa,L.   201 Fessas,D.   89
Esquerre,C.  911 Fevzioglu,M.  1321
Estrade,P.   843,2227,2233 Fica,X.   1719
Esvelda,E.   303 Ficzek,G.   1115
Euring,F.   1481 FidelGutiérrezLópez,G.  1045,1393,1999
Evageliou,V.  863,1273,2055 Filipoviđ,J.S.  1897
Evans,J.   2241,2245,2249,2251 Filipoviđ,N.K.  1897
Exarhopoulos,S.  693 Filipoviđ,V.S.  1897
Eze,S.O.   239 Filliol,C.   649
Fabbri,A.   1423 FinardiFilho,F.  1757
FabelaͲMorón,M.F.  1287 Fitry,I.   1373
Faber,T.J.   53 Fleurot,O.   2223
Fabiano,L.   2133 Flick,D. 55,125,295,523,535,
Faille,C.   1557,715 595,1435,1443,1493,
Falguera,V.   1299 1495,1531,1841,
Fang,B.   1333 2247
Fang,F.   449 Flores,S.   979
Farfán,M.   1079 Floros,J.   105
Farhoosh,R.  903,1093 Foerst,P.   1801
Faria,J.T.   1107 Foisy,M.   1261,2033
FariaSoares,C.M.  2131 Fokialakis,N.  2215
Farias,C.   1977 Follegatti–Romero,L.A.  1389,1391
FariasdeFarias,D.  1161 Fonseca,F.   267,561
Farid,M.   1601 Fonseca,I.M.  1023
Farina,A.   895,1177,2237 Fonseca,J.   1553
Farroni,A.   151 Fonseca,M.  793
Fasseas,K.   2105 Fontan,R.C.I.  1487
Fatnassi,S.   965 Forsberg,F.   593
Faulstich,M.  757 Först,P.   149,957
FavaroͲTrindade,C.S.  23,883,2165 Föste,H.   701,703
Favier,R.   965 Foster,K.D.   633
Fayolle,F.   713,1249 Foucquier,J.  323
FazaniCavallieri,Â.L.  933,1309 Fougère,L.   809
Fazheng,R.   2037 Fox,M.   723
Fazli,F.A.   1609 Fragkiadakis,G.A.  1835,1837
Fazli,N.A.   1609 Fragoso,E.   1817
Fehaili,S.   483,1805 Franca,A.S.   1693,1761
Felipe,A.M.P.F.  1019 Franchi,M.A.  1661,1663
FelixdeSantana,R.  2131 Franke,K.   569
Fencl,J.   1591 Franke,S.   731
Fenelon,M.A.  577 Fras,A.   2149
Feng,H.   411,1699 FraschͲMelnik,S.  695
Fereshteh,D.N.  1509 Freire,C.M.A.  1019
Feriotti,D.G.  1583 Freire,F.B. 333,1353,1361,1489,
FernandaZotarelli,M.  579 1569,1571
Fernandes,A.  103 Freire,J.T. 333,1353,1361,1489,
Fernandes,F.A.N.  1987 1569,1571,2005
FernandesNassar,S.  861 Freitas,D.DeG.  1225,1967,2151
Fernandez,A.  31,1237 Freitas,O.   2165
Fernández,K.  2101,2103 FreitasMoraes,I.C.  861,1179
FernándezͲMuñoz,J.L.  417 Frías,J.M. 285,453,773,1465,
FernándezͲSalguero,J.  2025,2027 1773,2209
FernándezͲSegovia,I.  1483 Fröhling,A.   393,1643
FernándezͲValle,M.E.  245 Fruilo,M.   349
Ferng,L.H.   1913 Frydenlund,F.  1539
Ferragut,V.   429,1193,1767 Fryer,P.J. 49,147,265,329,533,
Ferrando,M.  651,2193 621,697,711,913,
Ferrari,G. 165,349,371,681,1657, 915,1511,1543,1555
1729 Fuchigami,M. 1665,1667,1679,
Ferreira,A.R.  1023 2019
Ferreira,N.J.  1669 Fuchs,L.   587
Ferreira,R.G.  2093 Fuentealba,C.  627

I-8
Fuentes,A.   1483 Gioielli,L.A.   1047
FuentesBerrio,L.  1219,2017 Giovanis,T.   1847
Fujihara,E.   2119 Giuliani,R.   1933
Fujii,T.   369 Gloria,M.B.A.  1781,2065,2111
Fujita,K.   455,1281,1883 Gobbi,S.   1577
Fukuoka,M.  319,1917,1925 Gochangco,M.U.  1929
Fundo,J.F.   963 Godoy,H.T.   2179
Furtado,A.A.L.  1225 Godoy,S.   2101
Furuta,T.   7,907 Gogou,E.   1653
FyllingͲJensen,Ø.  2223 Gohari,A.R.  893
Gabas,A.L.   735 Gök,S.B.   791
Gabsoo,D.O.  1145 Gökmen,V. 485,1039,1581,1755,
Galán,E.   2025,2027 1807,1825
Galanaki,P.   863 Golding,M.   185
Galanakis,C.M.  737,1501,1579 Goli,T.   1349
Galier,S.   385 GoliMovahhed,G.  1907
Galoburda,R.  563,1199,1201,1217,2199 Golota,V.I.   575
Galúcio,C.S   2063 Gomes,A.   665
Galvez,C.   167,1677 Gomes,D.   1941
Gamboa,O.Diaz  2197 Gomes,F.S.   1641,2203
Gao,Z.ͲJ.   2013 Gomes,G.V.L.  19
Garbelotti,M.L.  1189,1375 Gomes,M.Z.V.  1549
Garcia,A.O.   2145 Gomes,S.   665
Garcia,L.C.   839,841 Gomez,P.L.   1635
Garcia,M.R.  281 GómezGalindo,F.  515
GarcíaMelón,M.  661 Gomide,C.A.  1179,1329
GarcíaͲPérez,J.V.  1951,1997 Goncalves,E.  1033,1647
Garczarek,U.  505 Gonçalves,C.  2151
GaribayFebles,V.  1999 Gonçalves,J.C.  2061
GaridoͲFernandez,A.  2211 GöncüoŒlu,N.  1825
Gaspar,A.   1595 Gondek,E.   1147,1385,2235
GasparMoreno,J.  747 Goñi,S.M.   1425,1803
Gaston,E.   285,1465,1959 GonzalesͲBarron,U.  473,497,1819
Gatellier,P.   1855 Gonzalez,J.E.  523
Gaucel,S.   323 González,K.  961
Gaukel,V.   949 González,R.  1277,2027
Ge,Z.   1123 GonzálezMariaPatricia,C. 1197
Gebhardt,R.  13 GonzalezͲMartínez,C.  217,1335,1963
Gedrovica,I.  2149 Goodman,P.A.  1593
Geeraerd,A.H.  475,2241,2245,2249,2251 Goossens,J.  475
Gekas,V.   737,1501,2207 Gordon,P.W.  709
Gelaw,T.K.   2193 Gorgen,M.   1643
GélvezOrdóñez,V.M.  1219,2017 Goto,C.   2201
Georgantelis,D.  1209 Goto,K.   91,127,1541
Georgiadou,M.  163 Goto,M.   691.2201
Gerkema,E.  249 Gougeon,R.  373
Gerolemou,A.  1273 Gougouli,M.  1787
Geyer,M.   1643 Goujot,D.   311,1867
Ghai,G.   529 Goula,A.M.   571,1517,1809,1957
Gharachorloo,M.  1297 GoulartMachado,V.  2095
Ghiaus,A.ͲG.  1899 Goulas,V.   1501
Ghikas,D.   495,1765 Gounadaki,A.S.  1829,1831
Ghiselli,G.   401 Gowen,A.A.  285,911
Giampaoli,P.  47,483,1805 Graciela,A.   2249
Gianfrancesco,A.  243,1137,1801 Graef,V.   21,121
Giannini,A.N.  2077 GramChristensen,M.  269
Giannoglou,M.N.  1851 Gramatina,I.  563
Giannopoulos,D.  849 Gras,M.L.   2089,2091
Giannou,V.   1775 Grassi,M.   895,1177,2237
Giannoulis A.  849 Grauwet,T.   351,367
Giardiello,M.  243,1137 GrayͲStuart,E.M.  337
Gibis,D.   1003 Greiner,R.   21,121
Gil,B.   1459 Grenier,D.   125,1435,1443
Gil,M.   1941 Grigoroudis,E.  1869
Giner,S.A.   549,2041 Grimaldi,R.   1047,1049,1293

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-9
Grinde,M.   511 Hampel,R.   649
Grob,F.   569 HamzalioŒlu,A.  485
Groeneveld,R.  2225 Han,Q.   993,1983
Grogan,H.   1773 Han,Wu   1673
Grogg,P.   1637 Hancocks,R.D.  1745
Gross,D.   2233 Hannesschläger,G.  1175,2229
Grossiord,J.ͲL.  1249 Hansriwijit,S.  533
Grosso,C.   1139 Hardacre,A.  669
Grosso,C.R.F.  1267,1387 Harnkarnsujarit,N.  1127
Grosso,R.C.  2197 HarrardeDienes,A.  661
Grupa,U.   1481 Harsa,b.   219,1039,2043
GruzdienĦ,D.  1271 Hartel,R.   87,727,779,1213
Gryz,K.   1255 Hashimoto,A.  1863
Gryz,S.   1253,1255 Hashitani,N.  1721
Grzegorzewski,F.  423 Hashizume,K.  127
Grzesik,M.   1251 Hassan,M.A.  233
GuadalupeSosaͲHerrera,M. 837 Hassanzadeh,A.  241
GuadarramaFernández,L. 1045 Hauser,C.   107
Gualdron,L.  1173 Havet,M.   339
Guamis,B.   429,1193,1467,1767 Hayakawa,S.  91
Guanoquiza,M.  1677 Hayashi,M.   369
Guardiani,V.  2263 Hazizaj,A.   421
GuastaferroSeravalli,E.Ap. 1757 He,J.   779,1213
GuedesͲdeͲPinho,P.  1861 Hebbink,G.A.  51
Güell,C.   651,2193 Hedreira,J.   2109
GümüƔ,B.   1453,1455 HeiluFayissa,A.  269
GuenterKieckbusch,T. 1007,1017,1047,1049, Heinemann,R.J.B.  23,2165
1065,1293 Heinz,V.   569
GuerraRodríguez,M.E.  1413 Heldman,D.R.  251,307
Guerrero,S.N.  1311,1629,1795 Hellenas,K.ͲE.  1217
Gueven,A.   391,1151 Hendrickx,M.  351,361,367
Guignon,B.   205 Henehan,G.T.M.  2209
Guillemin,H.  1947 Henke,S.   1695,1707
Guiné,R.   1263,1265 Henriques,F.  1265
Guizani,N.   153,1119 Henriques,M.  1941
Gul,O.   1295 Heo,S.   2073
Gulzar,M.   1149 Heredia,A.   1993,1995
Gumy,J.ͲC.   57,649,845,2231 Hernández,H.M.  1999
Guney,F.   1597 Hernández ,H.  445
Gupta,M.   1651 HernándezPezzani,C.  1195
Gustafsson,K.  2047 HernándezSánchez,H.  445,1163,1393
Gut,J.A.W.   1347,1439,1445 HernándezͲBrenes,C.  2045
GutiérrezdePiñeresc,C. 2109 HernándezͲHerrero,M.  429,1193
GutierrezdePiɶeresRocha,C. 2107 Hernando,I.  1621
GutiérrezLópez,G.F. 1045,1163,1287,1753, Herppich,W.B.  1643
1999 Herremans,E. 139,209,843,1175,
GutierrezͲUribe,J.A.  2045 2227,2229,2239
Guven,G.   581,1931 Herrera,M.L.  923
Guzman,F.   615 HerreraͲPérez,L.  1221
Gwanpua,S.G.  2245 Heyd,B.   1921
GʊmezGalindo,F.  565 Hicsasmaz,Z.  1151
Haas,K.   1551 Hinkova,A.   1695,1707
HaddadKhodaparast,M.H. 1509,1689 Hinojosa,A.  1827
HadnaĜev,M.  881,1109,2141 Hiraiwab,Y.  1191
HadnaĜev,T.D.  881,1109,2141 Hirano,M.   1033
Hadwiger,K.  765 Hoang,M.   1841,2247
Hafner,A.   1203 Hodara,K.   1251
Hagiwara,T.  313,525,1561 Höfler,J.   731
Hagura,Y.   155,1121,1303 Hogan,S.A.   1379
Håkansson,A.  587 Hojjatoleslamy,M.  1297
Halik,M.   1253 Holah,J.   467
Hamada,N.   1917 Holzmüller,W.  13
Hamaker,B.R.  1321 Hong,S.ͲI.   989
Hamanaka,D.  1859 Hook,B.   229
Hamberg,L.  539 Hooley,P.   1043

I-10
Horabik,J.   1759 Jaques,A.   1521
Hori,M.   1187 Jaques,D.   1521
Horziđ,D.   865,867,2039 Jara,F.   1141
Hosono,M.   1743 JaramilloͲFlores,M.ͲE.  961,1163
Hotrum,N.   723 Jardim,D.C.P.  1799,2145
Houška,M.   639,1165 Jauregi,P.   377,395
Hsu,C.ͲL.   1461,1565,1567 Jay,F.   2249
Hsu,K.ͲC.   2009 JeLee,S.   1153
Hu,H.   441 JeanͲPierre,P.  489
Hu,W.   439 Jeelani,S.A.K.  635
Hu,X.   1617,1659 Jekle,M.   25,79,885
Huang,Q.   947 Jelicich,A.   1381
Huang,S.ͲL.   2125 Jianu,C.   1909
Huang,T.ͲS.  1871 Jianu,I.   1909
Hubinger,M.D. 839,841,853,1067,1267, Jibu,Y. 1665,1667,1679,
1269,1387,1991,2001, 2019
2187 Jiménez,A.   871
HuckͲIriart,C.  923 Jiménez,E.   1639
Hui,Z.   29 Jiménez,M.  1603,1639,2097
Hultin,H.O.   1955 Jiménez,M.T.  1791
Hultman,E.   593 Jiménez,N.   481
Humpfer,E.  2233 JiménezͲAguilar,D.M.  2045
Hung,Y.ͲC.   405 JimenezͲAparicio,A.  1163
Hurtado,S.   1239,1241 JiménezͲAparicio,A.R.  1287
Hussein,H.F.  531 JimenezͲJunca,C.  57
Hussein,M.A.  1473,1513 Jin,X.   249,1607
Hutin,A.   1443 JinWe,K.   1099
Iamanaka,B.T.  1747 JoãoBarroca,M.  1265
Ibarz,A.   1299 Johnsen,S.O  471
Ibrahim,A.H.  213,259,1697 Jones,J.R.   337
IgnacioAzocar,M.  1159 Jordan,K.   1829
Igoumenidis,P.E.  1341,2175 JosédoAmaralSobral,P. 37,861,883,1009,
Iguti,A.M.   1583,2059,2133,2135 1033,1143,1179,
Ilg,Y.   99 1329
Iliná,A.   389,1739 JoseMazziniSartori,D.  275,1367
Illanes,A.   1239 JoséRodríguez,M.  2049
Indergård,E.  511,2241,2245,2249,2251 Juan,B.   1767
Inès,B.ͲA.   489 Juhász,R.   2213
Ingram,A.   265,1543 Jun,S.   435
Innings,F.   539,587,593 Jung,J.   1099
Isaji,E.   1541 Jury,V.   199,1205
Ishida,N.   15,441,1051 JustoAlonso,M.  1539
Ishii,K.   1665,1667,2019 Kabbani,D.   1959
Ishiwatari,N.  1917 Kadlec,P.   1165,1695
Isleroglu,H.  581,1931 Kadoyanagi,T.  1859
ItoMorioka,L.R.  2137 Kajiwara,K.   1901
Ivanov,S.V.   847,1081 Kakali,F.   1843
Jablonka,O.  2149 Kakuda,P.M.  2187
JacobKurtz,D.  1839 Kalantzi,K.   495,1765
Jacolot,P.   415 Kalogeropoulos,N.  2177
Jaeger,H.   419,431,437,607 Kaltsa,O.   69
Jagus,R.J.   833,979,1691 Kaluševiđ,A.  173,2057,2143
Jahanbin,K.  893 Kameoka,T.  1863
JaimeͲFonsecaa,M.R.  621 Kamil,R.   309
Jakab,D.   2213 Kammerer,D.R.  617
Jakubczyk,E.  843,1147,1385,2227,2235 Kanchanatawee,S.  385
James,B.   211,247,1035,1601 Kang,C.G.   1575
James,C.   527,729,1903 Kang,S.   1771
James,S.J.   527,729,1903 Kang,T.   1123
JanvanderGoot,A.  203,719 Kapetanakou,A.E.  1885
JaneteAngeloniMarcon,M. 1839 Karapantsios,T.D.  537,919,1243,1927
Jang,H.D.   1461,1565,1567 Karásková,P.  1483
Jangam,S.   555 Karathanos,V.T. 257,583,813,1341,
Janssen,A.E.M.  719 2123,2175,2177
Jao,C.ͲL.   2009,2125 Karatzas,K.A.G.  409,427

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-11
Karbowiak,T.  373 Koakuzu,S.N.  2151
Kargar,M.   61 Kobayashi,I.  73,573,937
Karipoglou,C.  1843 Kobayashi,T.  1351
Karklina,D.   1217,2149,2183 KocadaŒlŦ,T.  1581
Karpinski,K.  469 Kohda,J.   283,1457
Karvela,E.   2177 Köhler,H.   707
Karwe,M.V.  363,1681 Köhler,K.   591
Kasahara,I.   1229,1637 Kokawa,M.   455,1281
Kasuya,S.   1187 Kokini,J.L.   5,45,889
Katapodis,P.  1683 Kokkinaki,A.  1835,1837
Katayama,T.  369 Kokkinakis,E.  1835,1837
Katikou,P.   1209 Kole,A.P.W.  505
Katina,K.   1331,113 Komaitis,M.  745,863,1273
Katsanidis,E.  1845 Komen,G.   219
Katsaros,G.  1683,2243,2245 Komes,D.   865,867,2039
Katsikogianni,M.G.  1001 Kometani,T.  1191
Kaufmann,S.F.M.  637 Kondjoyan,A.  1855
Kawabta,M.  1541 Kondo,S.   2201
Kawagoe,Y.  1901 Kong,F.   1381
Kawai,H.   121,1551 Konstanta,M.A.  1341
Kawai,K.   155,1121,1303 Konteles,S.J.  583
Kawanishi,H.  1145 Kontominas,M.G.  1209
Kawashima,M.  2119 Koroleva,O.  2261
KayaͲCeliker,H.  169 Koronaki,E.D.  97
KaymakͲErtekin,F.  581,1931 Kostoglou,M.  919,1927
Ke,X.   1895 Kostov,G.   1451
Kechichian,V.  1445 Kotlar,C.E.   1709
Keener,K.   427 Koudou,M .  1935
Kehlenbeck,V.  803 Koulis,M.   2029
Kelkar,S.   187 Koulouris,A.  1605
Kemerli,T.   581,1931 Koutinas,A.  399,745
Kerasiotis,T.  1021 Koutsoumanis,K.  101,463,1787,1788
Kerjean,C.   1749 KovaēeviđGaniđ,K.  2039
Kerry,J.P.   1943,1961 Kovacs,A.   499
Khan,M.A.   81,967,1207,1975,2195 Koyanagi,T.  1935
Khayat,S.   1249 Kozarski,M.  1383
Kholodova,E.A.  575 Krahenbühl,M.A.  1131
Khosravi,A.R.  1245 KrauseBierhalz,A.C.  1007,1017
Khutoryanskiy,V.V.  679 Kreicbergs,V.  2199
Kiani,H.   521,1319 Kreyenschmidt,J.  99
Kieckbusch,T.G. 1007,1017,1047,1049, Kreyenschmidt,M.  99
1065,1293,1747 Kristo,E.   421
Kihara,F.S.   1747 Kroh,L.W.   423,607
Kim,C.ͲJ.   823 Krokida,M. 257,1331,2077,2087,
Kim,C.ͲT.   823,1459 2123
Kim,E.   247 Krstonošiđ,V.  881
Kim,E.H.ͲJ.   633 Kruma,Z.   563,2183
Kim,M.S.   1771 Kucera,P.   639
Kim,N.   823,1459 Kudra,T.   491
Kim,S.ͲW.   1099 Kühnl,W.   375
Kim,T.ͲE.   1459 Kukurová,K.  1591,1881,1965
Kim,Y.J.   1575 Kulozik,U. 13,149,375,381,397,
Kimura,C.Y.C.S.  1131 675,685,957,1687,
Kimura,Y.   1665,1667,2019 1801,2099
Kiosseoglou,V.  129,879 Kumagai,H.  2119
Kirs,E.   1343 Kumar,Y.   1207
Klaus,A.   1383 KumarMallikarjunan,P.  169
Klocke,S.   393 Kunigk,L.   2093
Knight,G.   469 Kunkulberga,D.  1889
Knoerzer,K.  335,1417,1535 KuoͲChiang,H.  2125
Knorr,D. 355,357,391,393,419,431, Kurenda,A.   1763
437,607,777,877,891, Kurozawa,L.E.  1991
1633,1643 Kushiro,M.   1883
Ko,S.   831,1099,1575 Kusztal,R.   843,2227
Ko,W.ͲC.   2009 Kuwada,H. 665,1667,1679,2019

I-12
Kviesis,J.   1199 Leiva,M.   1737
Kyriakidis,G.  1835,1837 Leloup,M.   1355,1357
LaPenna,M.P.  179 Lema,P.   1005
Laackmann,J.  63 LemusͲMondaca,R.  1491
Labropoulos,A.  1921,1947 Lenas,D.   1843,1865
Labuza,T.P.   75,775 Lentzou,D.   1337
Lacenlotti,F.  1521 Leray,D.   199
Lafosse,M.   809 LeskošekͲukaloviđ,I.  2057
Lagarón,J.M.  2249,2251 Lesmes,U.   625
Laghi,L.   797 Lewis,W.   603
Lagorio,G.   151 Li,L.ͲT.   931
Lagrange,L.F.  733 Li,S.   993,1983,1985
Laguerre,J.ͲC.  415 Li,W.   2157
Laguerre,O.  1841,2247 Li,X.   795
Lalechou,A.  1847 Li,Y.   411,993,1983
Lalpuria,M.  105 Lianou,A.   1787
Lamardo,L.C.A.  1181,1189 Liao,X.   439,1659,1617
Lamrini,B.   171,1427 Liapis,A.I.   77
Lancellotti,F.  541 Lienemann ,K.  2225
LandiFranco,C.M.  225 LiewAbdullah,A.G.  387
Langwoski,H.ͲC.  731 Lijun,C.   2037
Lanoisellé,J.ͲL.  909,1939 Lim,J.ͲG.   1575,1771
Lanza,M.   1389,1391 Lima,Á.S.   1703,2131
Laohasongkram,K.  677,1613 Lin,T.   1123
Laos,K.   1343 Lin,Y.A.   1913
Laparra,J.M.  217 LingTsai,C.   1281
LaraGuzmán,O.  2015 Link,M.   449
Laridon,Y.   125,1443 Lioumbas,J.S.  537,1927
Larue,O.   1215 LirenyGuaraldoGonçalves,A. 2197
Lasagabaster,A.  353,741 Lite,Li   1895
LatorreSalamanca,L.E.  1027 Littlejohn,K.A  1043
Lauhon,R.   195 Liu,D.   459,1939
Lavenson,D.M.  901 Liu,L.   287
Lazarides,H.N.  753,789,1517 Liu,S.   319
Lazaridou,A.  143,2207 Liu,Y.   383
Lazou,A.   2123 Livney,Y.D.   683
LeGot,A.   633 Llave,Y.   313
LeLouët,L.   1855 Llorca,I.   1563
LePage,J.ͲF.  965 Llorens,A.   31,1237
LeRévérend,B.J.L.  147,915,1887 Lloret,E.   31,1237
LealJr.,W.F.  1967 Lo,Y.M.   567
Leaungvongpaisan,G.  659 Locatelli,G.   2085
LeͲBail,A.   199,1205 Loginova,K.  1623
Lebesi,D.   1345,1775,1783 Londoño,J.   1777,2015
Lebovka,N.   403,909,1623 LoonNeoh ,T.  7,907,991
Leducq,D. 523,1531,2241,2245,2249, LopesdaCunha,R.  933,937,941,1309
2251 López,J.   2049
Lee,C.   1833 López,J.A.   399,745
Lee,H.   411 LopezGarcia,I.  399,745
Lee,H.ͲH.   989 LópezMa.Reyna,R.  445,689,1617
Lee,I.   1099 LópezͲCervantes,J.  1157,2011
Lee,J.W.   411 LópezͲMalo,A.  1791
Lee,K.   1771 LópezͲQuiroga,E.  805,1409
Lee,P.S.   831 LoredoGarcia,A.B.  1311
Lee,S.   1575 Lorén,N.   783
Lee,S.H.   435 Lorenz,R.   99
Lee,S.M.   2073 Lorenzo,G.   137
Lee,W.   717 Loss,R.D.   1447
Leeb,E.   397,685,2099 Lou,Z.   2013
Legentilhomme,P.  713 Loukianou,M.  1851
Lehtinen,P.   2105 Louppis,A.P.  1209
Leiqing,P.   1123 Lourenço,L.H.  1227,1233
LeiteMittererͲDaltoé,M. 1161 Loures,M.M.R.  1107
Leitner,M.   845,1175,2229,2231 Loveday,S.M.  131
Leiva,G.   279,289 Lu,P.   431

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-13
Lucas,T. 125,199,227,1435,1443, Martin,P.J.   229,1061
1475 MartinͲBelloso,O.  2217,2219
Luccas,V.   1293 MartínTorrezIrigoyen,R. 549
LucenaBarbosa,J.  2001 Martinez,P.  1655
Lule,I.   501 Martinez,R.  961
Luna,G.   1369,1603,1639 Martinez,V.  1563
LunaͲSolano,G.  2097 Martínez,C.L.  1257,2115
Lund,D.B.   751,767 Martínez,E.  2049
Lustre,A.O.   1929 Martínez,H.J.L.  389
Lutton,E.   325,1529 Martínez,J.L.  1739
Luyten,J.M.J.G.  53 MartínezdeMarañón,I.  353,741,1475
Lykidou,A.   693 MartínezPadilla,L.P.  837,1111
Ma,J.   993,1983 MartínezͲBustos,F.  2011
Ma,X.ͲX.   211 MartinezͲManez,R.  181
Machmudah,S.  691,2201 MartínezͲRivas,A.  1045
Mack,S.   1513 MartínezͲVelarde,R.  1287
Madrona,G.  1711 Martins,E.   1161
Maeda,A.   1721 Martins,J.T.  293,1015
Maeda,Ryo  1743 Martins,V.M.  1161
Maeng,J.ͲS.  1459 MartinsJacinthoBarbosa,M.I. 2001
Maffi,D.   2075 Martverk,K.  1343
MagañaͲBarajas,E.  1157 Mary,G.   195
Magee,T.R.A.  235,253 Mascheroni,R.H.  613,1441
Maghsoudlou,Y.  1497,1499 Masi,P.   263,1211,2069
Mahadevan,S.  1681 Masot,R.   1483
Mahajan,P.V.  971,1699,1741 Mastrocola,D.  2205
Mahamaktudsanee,T.  677 Mathilde,C.  489
Mahjoob,M.  1779 Mathys,A.   355,357
Mai,L.   2157 Matiacevich,S.  141,1087,1159,1183
Maifreni,M.  1731 Matser,A.   367
Majschak,J.ͲP.  701,703,707 Matsuda,A.  441
Makhonina,M.  1081 Matsui,H.   2119
Makinoc,Y.   1901 Matta,V.M.  1437,1641,1967
Makishi,F.   735 Mauermann,M.  707
Makita,G.T.  1725 Maureira,H.  1675
Makri,E.   1853 Mavragani ,I.  863
Makris,D.P.  2177 Mawatari,T.  1303
Malacrida,C.R.  2173 Mazloumi,M.T.  981,1185
MalikOthman,A.  1555 Mazutti,M.A.  1037,1125
Mallides,K.   359 McCarthy,K.L. 277,855,899,901,
Malone,C.R.  175 1155
Mandala,I.   69,947,2055,2105 McCarthy,M.J.  277,855,901,1155
Manios,S.G.  1769,1847 McClements,D.J.  625,929
Mank,V.   1059 McHugh,T.H.  215,795
Manojlovic,V.  173 McKenna,B.  605,787
Manzocco,L.  177,1731 MedeirosHirata,T.A.  2095
MarangonideOliveira,G. 1065 Medina,V.   1977
Maraschin,M.  1839,2179 Medina,W.T.  193
Marciano,E.  1375 Mediouni,H.  681
Maresca,P.   371,1657 Mehryar,L.   241,1713
Margas,E.   467 Meijer,H.E.H.  53
MariaZanin,G.  1235 Meinders,M.  303
Mariano,J.   649 Meireles,M.A.A. 821,1037,1125,1519,
MariuszPieczywek,P.  9 2113,2167,2189,
Markaki,A.   1835,1837 2191
Markman,G.  683 Meirelles,A.J.A. 1131,1133,1389,
Marková,L.   1591,1881,1965 1391,1405,1419,
Markoviđ,K.  2039 1701,1953
Maroulis,Z.B.  813 Mekhloufi,G.  1249
Marques,D.  2085,2137 Melado,A.   245,845,2231,2233
Marques,L.G.  2003,2005,2007 MeléndezͲPérez,R.  1129,1221
Marra,F.   629,1005,1945 Mello,B.C.B.S.  2187
Marshall,S.H.  615 Melnyk,O.   1059
Marson,G.C.  2145 Mendes,R.   827
Martin,C.   229 MendesdeFigueiredo,H. 2153

I-14
MendesdeFigueiredoJúnior,A. 2153 Molina,P.   141
Mendez,S.   1289 Monahan,F.J.  1001
Mendo,S.   1879 Mondor,M.  2033
Mendoza,J.  1325 Mondragón,G.  279,289
MendozaMadrigal,A.G.  623 MonroyͲVillagrana,A.  1163
MendozaPérez,J.A.  1999 MonspartͲSényi,J.  2213
Meneau,F.   119 Monteau,J.Y.  1205
Menegalli,F.C.  1009,2095 Monteiro,F.S.  551
Meneses,N.  437 MonteiroAndradeBarreto,I. 2031
Menkov,N.   1083 Montel,M.ͲC.  1947
MerazTorres,L.S.  1163,1393 Montes,E.   1325
MercadoͲMárquez,C.  1129 Montes,F.J.M.  1619
Mercer,D.G.  751,755,767,1371,1593 Montoya,D.  761
Mercier,S.   1261,2033 Moongnarm,A.  337
Merkouri,S.I.  1829 Moraes,F.   37,917
Mery,D.   279,289,1753 Moraes,I.C.F.  1033,1329
Mescher,A.  63 Moraes,I.V.M.  1641
Mesmoudi,S.  325,1529 Moraes,M.   27
Mession,J.ͲL.  875 Moraga,N.O.  1491
Meyer,X.M.  311,483,1867 Morales,E.   829
Meza,B.E.   835 MoralesͲdelaPena,M.  2217,2219
Mezdour,S.  195 MoralesRosas,I.  2011
Mgaidi,A.   681 Moreau,M.  1615
Michel,M.   687,2211 MoreiraDutraAlbuquerqueE.Silva,I. 2181
Michihata,T.  1935,2119 Moreno,C.   1073
Michon,C.   69,227,1443 Moreno,J.   1973
Mihailov,M.  1451 Moreno,M.C.  123,1077
Milczarek,R.R.  215 MorenoͲLucas,B.  1477
MillánͲCarrillo,J.  1157 Morgernstern,M.P.  247,337,633
Mills,T.   817 Mosca,A.C.   611
Milutinoviđ,M.  2057 Moschakis,T.  129,143,879,1055
Min,B.   1575 Moscovici,A.M.  183
Minemoto,Y.  1191 Mosnier,J.ͲP.  427
Ming,L.   1123 Motallebi,A.A.  981,1185
Minim,L.A.   1107,1705 MottinDemiate,I.  1839
Minim,V.P.R.  1107,1705 Moughan,P.  631
Mintzas,D.   693 Mourão,C.A.  1705
Mirade,P.ͲS.  965,1947 Mourtzinos,I.  2175
Miranda,G.   1277 Mousavi,S.M.E.  951,1319,1751
Miranda,M.  1677,2049 Mousavi,Z.E.  1319
Miri,T.   49,913 MoussaͲAyoub,T.E.  607,1091,1979
Mirkoviđ,N.  2083 Mousset,S.   2257
Mironescu l,D.  1105 Moutsatsou,P.  1021,1775
Mironescu,M.  1105,1377 Moya,S.   1475
Mistriotis,A.  849 Mozina,S.S.  769
Mitrea,D.   1401 Müller,J.   819,955
MiwaIguti ,A.  1757 MüllerͲBuschbaum,P.  13
Miyawaki,O.  1969 MujicaͲPaz ,H.  2139
Mizuno,T.   1721 Mujumdar,A.  555
Mödinger,L.I.  1715 Mulet,A.   1277,1951,1997
Moerkens,A.  203 Muniz,E.P.   1447
Moggridge ,G.  835 Muñoz,I.   1415
Moghaddam,T.M.  1279 Muñoz,K.   1777
Mogol,B.A.   1825 Muñoz,L.A.   985
Mohamad,N.L.  387 Muñoz,O.   627
MohammadiMoghaddam,T. 1247 Muranyi,P.   699,1551
Mohammed,I.K.  191 Murata,L.K.Y.  1439
Mohammed,M.A.P.  189,1275 Murata,N.   2119
Mohebbat,M.  1509 Murniece,I.  1217
Mohebbi,M.  1279 Murphy,E.G.  577
Moini,S.   893 Murray,A.   733
Mokhtar,A.  1473 Mussida,A.   473
MoldãoͲMartins,M.  673,793,1647,2061 Mustafa,L.   653
Molina,G.   1725,1727 MustapaKamal,S.M.  387
Molina,J.I.   281 Muthukumarappan,K.  597

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-15
Mytilinaki,E.  1629 Norton,I.T. 35,61,645,695,817,
N’guessanGeorges,A.  85 1745,1887
Nabetani,H.  455 Norton,T.   2157
Nabizadeh,F.  1167 Nótin,B.   1115,2213
Nagano,H.   1187 Nounesis,G.  1683
Nahar,S.   635 Nualkaekul,S.  1793
NairneNegrãoMurakami,A. 2181 Numthuam,S.  995
Nakagawa,K.  519,2171 Nunes,L.   827,1595
Nakai,T.   1925 Nuñez,H.   1407,1973,1977
Nakajim,M.  931 Núñez,G.A.   1715
Nakajima,E.  1191 NúñezͲSantiago,M.C.  1315
Nakajima,K.  369 NurHanani,Z.A.  1943,1961
Nakajima,M.  73,937 Nychas,G.ͲJ.E.  461
Nakamura,K.  1665,1679 O´Connell,C.P.  285
Nakamura,S.  1935,2119 O’Callaghan,D.J.  1379,1467
Nakano,L.A.  1967 O’Donnell,C.  911,1651
Nakano,Y.   283,1457 O’Gorman,A.  453
Nanasaki,Y.  1561 O’Neill,F.T.   1001
Narain,N.   2007 O’Sullivan,M.  1001
Nascimento,L.A.S.  1703 Obiedzinski,M.  1889
NascimentoeSantos,D.  1669 Occhino,E.   1621
NassiriMahallati,M.  1093 Oehlke,K.   121
Nasution,D.A.  477 Ogawa,T.   1351
Nathanailides,C.  1843,1865 Ognean,C.F.  1095
Natividad,A.C.  1929 Ognean,M.   1095
NavaGalicia,S.B.  1587 Ograsici,E.   1627
Nazir,A.   65 Oh,S.ͲM.   823
Ndoye,F.T.   295,535,1493 Ohmoto,E.   525
Nedoviđ,V.   173,825,2057,2143 Oikawa,S.   1303
Neidhart,S.   617 Oikonomopoulou,V.P.  257
Neill,G.   235,253 Oikonomou,N.A.  1331
Nemeth,A.   1175,2229 Okabe,S.   1541
Neri,L.   2205 Okai,K.   2201
Neves,M.A.  73,937 Okai,Y.   2201
Ngadi,M.O.  239,287 Okos,M.R.   717
Nguyen,H.H.  831 Okos,M.   187
Nguyen,L.T.  435 Oliveira,A.C.  2165
Nguyen,M.H.  557 Oliveira,A.L.  809
NguyenDoTrong,N.  139,209,1057,2239 Oliveira,F.   1741
Niazmand,R.  903,951 Oliveira,J.   1975
Nicolaï,B.M. 139,843,1057,2227,2239, Oliveira,J.V.  1037,1125
2245,2249,2257 Oliveira,L.S.  1693,1761
Nicolau,N.   1467 Oliveira,N.   1703
NicolettiTelis,V.R.  225,2173 OliveiradeFariña,L.  1823
Nicoli,M.C.   177 OliveiradeMoraesa,J.  953
Nicorescu,I.  1615 Olivera,D.F.  1785
Nielsen,B.   1533 Olivier,K.   85
Nieto,A.   1251 OlivieradosSantos,A.  1049
Nieto,G.   1469 Olsen,A.   897
Niksic,,M.   1383 Olsen,Ø.   471
Ninios,A.ͲI.   2105 Oprean,L.   1105
Niranjan,K.   57,655,1431 Orange,N.   1615
Nishi,M.   2119 Oreopoulou,V.  1585,1783,2053
Nishimoto,N.  519 ØrnholtZammit,G.  517
Nishizu,T.   91,127,1541 Orse,A.G.   1799
Nitschke,M.  1549 Ortega,J.A.A.  1619
Nitz,M.   2135 Ortiz,J.   1737
Niwata,E.   15,1051 Ortiz,M.E.   1079
Noguchi,A.   441 Ortiz,S.   1795
Nojiri,Y.   1191 OrtizͲPortilla,S.  1675
NordinIbrahim,M.  309 Oshitac,S.   1901
Nordlund,E.  2105 Osorio,E.   1777
NorneWidell,K.  1539 Osorio,F. 67,141,905,935,
Norton,A.B.  645 1075,1087,1183
Ospino,H.   1817

I-16
OuldAhmedou,S.A.  339 Peinado,I.   1993,1995
Ozcakmak,S.  1891,1893 Pelissari,F.M.  1009
Ozdestan,O.  581,1931 Penbunditkul,P.  255
Ozerov,O.   1645 Penha,E.M.  1967
Ozolina,V.   1889 Pénicaud,C.  1867
Oztop,M.   855 Penteado,A.L.  1225,1967
Ozuna,C.   1951,1997 Peralta,J.M.  709
Ozyurt,B.   581,1931 Perazzini,H.  1569,1571
P.Singh,R.   251,513,619 Pereira,A.C.I.  2133
Paez,M.   1159 Pereira,A.G.M.  1367
Pagani,M.M  2203 Pereira,A.L.F.  657
Paik,S.ͲY.ͲR.  831 Pereira,C.   1941
Painter,P.   889 Pereira,L.M.  1067
PaivaͲMartins,F.  917 Pereira,R.N.  413
Palazoglu,K.  1755 PereiraQuadros,C.  2083
Paleologos,E.K.  1209 Peresichnyi,M.I.  2035
Pall,R.   1343 PeretͲAlmeida,L.  2111
Palma,B.   2185 Pérez,L.M.   973
Palou,E.   1791 Pérez,O.A.   1325
Palzer,S.   197,637 Pérez,O.E.   407
Pan,Z.   795 PérezCabrera,L.E.  1027,1963
Panagou,E.Z.  461,2211 PérezMasiá,R.  2251
Panarese,V.  797 PerezͲEsteve,E.  181
Panayotov,P.  2159 PérezͲGonzález,J.  1025
Pangloli,P.   405 PérezͲWon,M.  1677
Pani,P.   1937 Peres,C.   2211 
Panopoulos,S.  1843 Perone,N.   1211
Papadaki,I.   1783 Perret,B.   1947
Papadakis,S.E.  583,2175 Perrot,N. 321,323,325,1427,
Papadimitriou,V.  2127 1529
Papadopoulos,T.  1683 Perunoviđ,M.  1285,2083
Papadopoulou,O.S.  461 Petermeier,H.  731
Papanikolaou,S.  745 Petroviđ,T.   825
Papaspyridi,L.ͲM.  2215 Petrušiđ,M.  2083
Papathanassiou,M.  891 Phan,V.ͲA.   505
Papavergou,E.J.  1857 Phoon,P.Y.   515
Papoti,V.T.   1243 Piazza,L.   599
Pappa,A.   2087 Piccone,P.   2081
Pappas,C.   2055 Pickardt,C.   617
Parada,J.   781 Picouet,P.   31,1237
Parada,L.   961 Picque,D.   1947
Paramera,E.I.  583 Pietsch,A.   601,1989
Parés,D.   1239,1241 Pifferi,A.   895,1177,2237
Park,K.J.   1991 Pighin,A.   1195
Pascual,A.   1563 PilarDoradoPérez,M.  399,745
PassaroCarvalho,C.  2015 Pilli,De   179,1933
Passot,S.   267,561 Pilosof,A.M.R.  407,833,1141
Pastore,G.M. 401,739,1589,1725,1727, Pimentel,J.D.R.  2007
2107,2179 Pimentel,M.R.  1725
Pastukhov,A.  1431 PinedaͲGómez,P.  1135
Pataro,G.   349,1729 Pinheiro,A.C.  293,673,1015
Paterson,A.H.J.  51,815,1283,1373,1429 Pinheiro,J.   1647
Patil,S.   409 Pinho,C.R.G.  1661,1663
PatriciaMartínezͲPadilla,L. 837,1111 Pinho,S.C.   19,27,747
Patsioura,A.  2207 Pino,A.   2025
Paunoviđ,D.  2083 Pinto,C.   1971
Paviani,L.   2071 Pinto,M.   541,1521
PavieCardoso,L.  1049 PintoKempka,A.  1573
Payne,F.   317,509,1469 Pirani,M.   2249
Paz,J.E.   299,1515 Pittia,P.   1621,2081
Pazir,F.   791 PlanaͲFattori,A.  295,1493,1495
Pecic,S.   2143 PlattͲLucero,L.C.  2011
Pedreschi,F.  67,905,935 Plessas,S.   1013,2029
Pedroso,D.L.  23 Poliseli,F.   429,1193
Peglow,M.   649 Polissiou,M.  2055

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-17
Polovka,M.   1171 Rafiee,M.   265,1543
Polychniatou,V.  943,1775 Rahman,N.N.Ab.  213,259,1697
Pontes,D.F.  2153 Rajabipour,A.  1779
Poonnakasem,N.  1613 Rakcejeva,T.  1199,1201
Popov,V.   2261 Rakesh,V.   1449
PorrasͲParral,G.  49,913 Ramane,K.   2199
Porto,P.S.S.  1447 Ramaswamy,H.S.  261,531
Portsch,A.   35 Ramírez,A.   959,1739
Potes,N.   157,785 Ramírez,C.   2109
Pouliot,Y.   1723 RamírezͲWong,B.  1157,2011
Pour,V.   1695,1707 Ramos,M.G.M.  1619
Poutanen,K.  133,1113,2105 RamosͲRiver,D.  1135
Prado,G.H.C.  1453 Rao,M.A.   131,811
Prado,J.M.   821,1519 Raphaelides,S.N.  693,1031
Prado,M.M.  1367,2003,2005,2007 Rasburn,J.   191
Prakashb,B.  593 Rashevska,T.O.  2023
Prazeres,P.   1949 Rashevskaya,T.  1081
Preechathammawong,N. 1063 Rashevskaya,T.A.  847
Pretti,A.A.   747 Raspor,P.   503,1547,1849
Prieto,W.H.  2135 Rastelli,S.L.   2081
Prokopiou,P.  1809 Raviyan,P.   411
Promeyrat ,A.  1855 Rayner,M.   59,2047,2079
Proshlyakov,A.  163 Razavi,S.M.A. 903,951,1093,1247,
Protonotariou,S.V.  2055 1611,1689
Provesi,J.G.  2181 Razborskaya,O.  1645
Proveti,J.R.C.  1447 Rega,B.   483,1805
Psaroudaki,A.  1869 ReginaKuhn,K.  1309
Psimouli,V.   2053 Reginatto,V.  1839
Psodorov,D.B.  1897 Reid,D.   185
Psomas,A.   1769 Reineke,K. 355,419,777,877,
Psomas,E.   1301 891
PsonkaͲAntonczyk,K.M.  1053 Reis,I.A.O.   1703
Ptaszek,A.   1253,1255,1313 Reis,M.H.M.  1711
Ptaszek,P.   1253,1313 ReisdaCosta,S.R.  1595
Puente,L.   1737,1971 Rejo,A.   477
Puligundla,P.  831 Relkin,P.   119
Pullawan,J.   2079 Remédio,L.N.  1143
Purchas,R.   1373 Remillard,P.A.  1593
Purroy,F.   365,1685 Renard,C.M.G.C.  201
Purwadaria,H.K.  457,477,547 RendónͲVillalobos,J.R.  1025,1339
Purwanti,N.  203 Rennie,T.J.   1371
Putier,F.   1355,1357 Requerre,A.ͲL.  133,1113
Putranto,A.  297,1505 ResendeSimiqueli,A.P.  401,739,1589
Qiding,Z.   1123 Reuillon,R.   325,1529
Qinghua,H.   1673 Revithi,A.   163
Quarta,B.   177 Revstedt,J.   587
Queiroz,M.I.  1161 Rey,I.   205
QueirozZepka,L.  1161 Rey,J.F.   1173,1257,2115
Quevedo,J.M.  1767 Reyes,J.E.   113,1677
Quezada,C.  1813 ReyesͲMoreno,C.  1157
Quinta,A.   37 Rezvani,E.   71
QuintaBarbosaBittante,A.M. 861,1179 Rhim,J.ͲW.   989
QuintanillaCarvajal,M.X. 1163,1393 Riaublanc,A.  323
Quintas,D.A.C.  293 Ribeiro,A.P.B. 1047,1049,1065,
Quintas,M.A.C.  293,963,1069 1293
QuinteroRuiz,N.A.  2041 Ribeiro,C.M.  1181
QuinteroͲRamos,A.  2011 Ribeiro,E.P.  2059,2093,2133
Quintino,A.  2085 Ribeiro,R.   747
Quispe,I.   2049 Ribeiro,S.C.A.  1227,1233
QuispeͲCondori,S.  2169 Ricci,J.   1349
RabeloTorres,E.  2131 Richardson,M.  609,1833
Radiđ,D.   825 Rieblinger,K.  1003
Raduloviđ,Z.  825,2083 RiekstinaͲDolge,R.  2183
Rafe,A.   1689,1907 Rigo,A.A.   1125
Raffellini,S.   1795 Rincon,C.   167

I-18
Ripberger,G.  815,1283 Saavedra,J.   1479,1813
Risum,J.   1533 SabancŦ,S.   2051
Rivas,D.   1537 Sabanis,D.   1775
Rivera,W.   167 Sabarez,H.T.  1503
Rizzolo,A.   895,987,1177,2075,2237 Sabovics,M.  563
Robbins,P.T.  711,1555 Sacchetti,G.  2205
Robert,P.   443,1717 Sacoda,P.   2071
Robin,F.   197 Sadd,P.   835
RoblesͲDelaTorre,R.R.  2021 Sadecka,J.   1965
RoblesͲLópez,R.  2021 Saeleaw,M.  869
Rocculi,P.   565,797,1011 SáenzCollins,C.  2121
Rodas,M.A.B.  1189,1375 Saeys,W. 139,209,1057,2245,
Rodrigues,C.E.C.  1701 2239
Rodrigues,D.  1941 Safonova,O.  969,575,1645
Rodrigues,E.  1397 Saghy,A.   1175,2229
Rodrigues,S.  657,1987 Sagis,L.   29,641
Rodrigues,DeMassaguerP. 1879 Sagis,L.M.C.  11,1041
Rodríguez,A.  613 SaŒlam,D.   11,1041
Rodríguez,J.  1739 Saguer,E.   1239,1241
Rodríguez,R.  1475 Saguy,S.I.   763,2217
RodríguezͲGarcía,M.E.  1135 Saito,E.   2071
RodríguezͲMarín,M.L.  1315 Sakai,N.   319,1917,1925
RodríguezͲSinobas,L.  245 Sakejha,H.   653
Roeckel,M.   2101,2103 SakinYilmazer,M.  581,1931
Rohn,S.   423,607 Sakiyama,T.  313,525,1561
RoisAnwar,N.Z.  147,915 Sako,H.   127
Rojas,J.   1463 Salazar,K.   1719
RojasdeGante,C.  2121 Saldaña,M.D.A.  2169
RojasͲGrau,Ma.A  2217,2219 SaleenaTaip,F.  309
Rolemberg ,M.P.  1131 Saleh,D.   653
Romani,S.   565,1011,1423 Salehi,F.   1611
Romano,A.   2069 Salmanian,B.  1245
Romero,S.   1817 Salmeron,I.  1793
Roos,Y.H. 145,157,577,785,925, Salova,A.   1695,1707
1943,1961 Salta,F.N.   1341
Rosa,E.   1993,1995 Salvadori,V.O.  1425,1785,1803
Rosales,A.   167 Salvatori,D.M.  1635
RosalesͲRiver,A.  1135 SalviaͲTrujillo,L.  2217,2219
Rosas,A.   1369 Sampaio,K.A.  1419,1953
RosasͲMendoza,M.E.  417,1129 SanJuan,E.M.  1929
Rosell,C.   199 Sanabani,AlͲ  1089,1981
Rosen,J.   1217 Sanchez,M.M.  1407
Rosenberg,M.  855 Sánchez,H.   445,1163,1393
Rosenthal,A.  1879 Sánchez,J.   1739
Rosentrater,K.A.  597 SánchezͲMachado,D.  1157,2011
Rosnes,J.T.   471,1789 Sancho,A.   1655
Rouaud,O.   1433,1545 Sanow,C.   1655
Roura,S.I.   1709,1811 Santacatalina,J.V.  1951,1997
RouxͲdeBalmann,H.  385 Santagapita,P.  797
Roversi,T.   599 Santana,I.Ap.  1757,2059
Rózek,A.   651 SantanaCastro,E.  2131
Rozos,G.   1013 SantéͲLhoutellier,V.  1855
Rubilar,M.   829,2185 Santos,A.L.  1225
Rubiolo,A.C.  973,975 Santos,C.T.   1097,1487
Ruiz,C.   2109 Santos,C.V.   2067
RuízCabello,J.  245 Santos,D.C.  1227
Ruktanonchai,U.  255 Santos,D.T.   2113,2191
Rumpf,I.   1203 Santos,I.P.   1447
Rumsey,T.R.  899 Santos,J.S.   1189
Rungchang ,S.  995 Santos,P.H.S.  135
Rungrodnimitchai,S.  659 Santos,S.D.   2067
Ruocco,G.   1945 Sanz,P.D.   205
Russ,W.   731,757,1481 Sanz,Y.   217
Rustad,T.   897 Sanzana,S.   2089
Rutherfurd ,S.M.  631 Sapci,B.   2051

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-19
Saraiva,J.   1879 Shigematsu,T.  369
Sarghini,F.   263 Shimoni,E.   17,117,183
Sartori,D.J.M.  1367 Shimoyamada,M.  1187
Sasaki,M.   691,2201 Shinzato,R.E.  1701
Sasaki,T.   1935 Shleikin,A.G.  663
Sastry,S.K.   433,1973 Shoji,A.S.   2165
Satake,T.   995 Shokohi,T.   1245
Sato,A.C.K.   1069,1071 Shukat,R.   119
Saucier,L.   1723 Shukla,R.N.  81,967
Saurel,R.   875 Shynkaryk,M.  433
Savoire,R.   1939 Sibakov,J.   2105
Sbaite,P.   2063 Siddiqui,M.  2195
Schäfer,H.   449,2233 Siddiqui,S.W.  945
Scharnhorst,K.  983 Siegert,N.   685
Schiavo,L.   461 Signorelli,M.  89
Schiraldi,A.   89 Sikes,A.   1833
Schirmer,M.  79 Silano,A.   263
Schleining,G.  71,869 Silva,A.M.   2067
Schlüter,O.   393,419,423,1633,1643 Silva,C.L.M.  479,771,963,1625
Schmitz,I.   1801 Silva,C.R.   27
Schoessler,K.  419 Silva,E.   641,2181
Schöler,M.   701,703 Silva,F.C.N.N.  1823
Scholl,S.   701,703,705 Silva,F.F.   1781
Schreurs,P.J.  53 Silva,F.V.M.  1601
Schröder,J.   63,949 Silva,L.F.M.  1641
Schroeder,B.  1655 Silva,L.R.   1861
Schroën,K.   65 Silva,M.C.   1553,1861
Schubert,H.  1 Silva,P.   1183
Schuchmann,H.P.  1,197,591,949 Silva,R.A.F.   2133
Schütz,B.   449 Silva,S.M.   1419,1953
Schulz,M.   431 Silva,T.C.A.   1821
Schvartzman,S.  1829 SilvaLima,Á.  1703
Schwartzberg,H.  159 Silveira,A.M.  1353,1361
Schwarz,K.   63 SilveiraJr,V.  1527
Seglina,D.   2183 Simmons,M.  533
Segura,L.A.   299,1365,1515 Simplício,I.A.S.  19
Seifzadeh,M.  981,1185 Simpson,R.J. 541,1407,1521,1973,
Semrau,J.   335 1977
Semyonov,D.  17 Singh,H.   131
SenglingLacerda,R.  1179 Singh,R.P. 161,251,513,619,
Sepulcre,F.   1959 629,631,799,1381
Sepúlveda,A.  1973 Siracusa,V.   1011
SerafiniPereira,C.  739 Sjöholm,I.   2079
Seravalli,E.A.G.  1757,2135 Sjolm,I.   1169
Serfert,Y.   63 Sjöö,M.   59
SermentͲMoreno,V.  1671,2139 Skaltsounis,A.ͲL.  2215
Serna,R.   1777 Skandamis,P.N. 1769,1829,1831,
Serra,A.T.   673 1847,1885
Sessa,M.   165,681 Skåra,T.   471,1789
Setinova,I.   639 Skondras,D.  1021
Seuvre,A.ͲM.  373,1919 Skudra,I.   563
Severini,C.   179,1933 Skurtys,O.   141,905,1087,1231
Seyis,F.   1891 Smith,B.   247
Sfakianakis,P.  1649,1775 Smith,J.   185,669
Shaffer,K.   51 Soares,C.M.F.  1703,2131
ShafiurRahman,M.  153,1119 Soazo,M.   975
Shahabi,I.   1751 Šobajiđ,S.   1285
ShahidiNoghabi,M.  903,951 Sobral,P.J.A. 37,861,883,1009,
Shalbart,P.   2249 1033,1143,1179,
Shamaei,S.   271 1329
Sharma,A.   487 Solliec,C.   1355,1357
Shatta,A.   1089,1979,1981 SolorzaͲFeria,J.  1025,1339
Shene,C.   829,2185 Soma,P.K.   567
Sher,A.   57 Somavat,R.   433
Shibata,M.   455,1281 Sommier,A.  1463

I-20
Soottitantawat,A.  171,192,2555 Sylla,Y.   715,1557
SosaHerrera,M.G.  837,1111 Symoneaux,R.  201
Sotiroudis,T.G.  2127 Szerman,N.  1195,1655
Soto,A.   1753 SzymaŷskaͲChargot,M.  1053
Soto,C.   2163 TabatabaeeYazdi,F.  1509
Souchon,I.   807 TabiloͲMunizaga,G.  1677
Sousa,I.   103 Tabuchi,M.   1665,2019
SousaͲGallagher,M.  1741 Tadini,C.C.   111,1445
Souza,A.L.R.  2203 Taherian,A.R.  71
Souza,C.   2137 Taipina,M.S.  1181,1189,1375
Souza,D.S.   2007 Taitano,L.Z.  161
Souza,R.A.   2063 Takahashi,C.  573
Sowasod,N.  519,2171 Takahashi,H.  1303
Soysal,A.   2251 Takatsu,K.   1541
Sozer,N.   45 Take,H.   2119
Spiess,W.E.L.  749,751 Takeiti,C.Y.   2151
Spigno,G.   165,377 Talens,P.   871
Spinelli,L.   895,987,1177,2243 Tambor,K.   1385
Spraul,M.   449,2235 Tan,J.ͲH.   1601
Spyropoulos,F.  35,61,645,695,1745 Tan,X.H.   1873
Stahl,M.A.   2063 Tanaka,F.   1859
Stahl,V.   2245 Tanaka,M.   691
Stajiđ,S.   1285,2083 Tang,J.   459
StåleNordtvedt,T.  1203 Taniguchi,H.  1935
Stavrakakis,E.K.  1957 Tanthapanichakoon,W.  519,2171
Stavropoulou,E.  1013 Tao,N.   383
Stavros,F.   1683 Taoukis,P.S. 359,559,891,1653,
Steele,R.   1749 1683,1851,1853,
Stefanakis,A.G.  1957 2241,2243,2245
StégerͲMáté,M.  1115,2213 Tarantili,P.A.  977
Steiner,I.   983 Tarantilis,P.A.  2055
Steinhauer,T.  375 Tarhan,Ö.   1039
Stella,S.   187 Tarleton,E.   189,1275
Stenby,M.   1533 Tashiro,M.   127
Stephan,M.P.  1225 Tassou,C.C.  461,2211
Stevik,A.M.  1905 TecanteCoronel,A.  1963
Stieger,M.   207,611 Tedesco,J.C.G.  19
Stoeber,M.   757 Teixeira,A.A.  529,541,1407
Stoeva,D.   1451 Teixeira,J.A.  413
Stoforos,N.G.  341,359,1797,1809 Teixeira,J.   1741
Stokke,B.T.   1053 TeixeiraCarvalho,O.  111
Storrø,I.   897 TeixeiraFreire,J.  1359
Strati,I.F.   1585 Tejada,A.W.  1929
Strautniece,E.  1201 Teles,A.S.C.  1641
Strixner,T.   1687 Telis,V.R.N.  1347
Stroebinger,N.  631 TelisͲRomero,J.  1347
Strohalm,J.   1165 Temelli,F.   443
Ströher,R.   1235 Teramoto,A. 1665,1667,1679,
Su,D.   1985 2019
Suehara,K.ͲI.  1863 Ternovskoy,G.V.  663
Sugimoto,Y.  1863 TerrésRojas,E.  623,1999
Sugiyama,J.  455,1281,1883 Terzi,S.C.   1641
Suhaj,M.   1171 Tessier,F.J.   415
Suhil,M.   547 Teymurova,A.  969
Sui,Q.   335 Tfouni,S.A.V.  1877
Sun,D.ͲW.   237,447,451,521,1631 Thaiudom,S.  1101
Suntudprom,J.  1153,1873 Thomareis,A.S.  1317
Surel,C.   323 Thomazini,M.  23,2165
Surowsky,B.  1633 Thompson,C.  509
Susukio,T.   1191 Thuault,D.   2245
Suthiluk,P.   995 Thürling,K.   109
Suzuki,T.   1187,1901 Thussu,S.   305 
Svanberg,L.  783 Timgren,A.   59
Svidlo,K.V.   2035 TiqueRaice,R.  1169
Swartzel,K.R.  545 Tiwari,B.   1651,2157

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-21
Tjahjohutomo,R.  547 Uquiche,E.   1719
To,N.   201 Uren,A.   581,1931
Tobin,J.T.   577 Uresti,D.N.   389
Tobolková,B.  1171 Ureta,M.M.  1785
Toepfl,S.   347 Uribe,E.   2049
Tokizawa,K.  525 Urrea,A.   1257,2115
Tokuda,K.   1935 Urrea,G.R.   1369
Toldrà,M.   1239,1241 Urrego,F.A.  1363
Toledo,M.C.F.  1875,1877 Uyar,R.   1411
Tommaso,G.  747 Uysal,R.S.   2051
Tonello,C.   365,1685 Uzelac,G.   2057
Tonon,R.V.   839,1267,1269,1387 Vaillant,F.   481
Topalidou,S.  1307 Valance,N.   1919
Torbica,A.   881,1109,2139 ValdezͲFragoso,A.  2139
Tornberg,E.  223,737,1305,1579 Valdramidis,V.P.  409,427,471,1789
Torreggiani,D.  1577,1937 Valencia,D.   429,1193
Torres,P.I.   1157 Valencia,P.   1815
Torres,R.   1325 Valentão,P.  1861
TorresͲChávezb,P.I.  2011 Valous,N.A.  451
Torricelli,A.  895,987,1177,2237 VanAs,H.   249,303
Torrieri,E.   1211,2069 VanAsselt,A.  723
Tóth,M.   1115 VanBoekel ,M.A.J.S.  505
Trägårdh,C.  587 VanBoxtel,A.J.B.  249,1607
Tréléa,I.C.   267,321,1427 VanDalenc ,G.  303
Trentin,A.   2193 VanDamme,I.  229
Trezzani,I.   689 VandeVelde,F.  611
Triantafillou,D.  1865 VandeWalle,D.  2233
Tribst,A.A.L.  1661,1663 VanderGoot,A.J.  203,719
Trindade,M.A.  2165 VanderPlancken,I.  351,367,2225
Trinh,L.   1061 VanderSluis,S.  2245,2249
Trinquet,F.   2249 VanderSman,R.  303,343
Troncoso,E.  921,929 VanderSman,R.G.M.  249,301
Trujillo,A.J.   1767 VanDerlinden,E. 11,641,1041,471,
TrujillodeSantiago,G.  2121 475,499,501,1789
Trystram,G. 171,227,321,689,1349, VanDeventer,H.C.  1607
1427,1435,1915 VanDuynhoven,J.  303
Tryvittayasil,V.  1859 VanHecke,E.  909
Tsatsaragou,K.  1851 VanImpe,J.F.M. 465,471,475,499,
Tsavali,G.   1307 501,1789
Tsekoura,F.  745 VanLoey,A.  351,367
Tsimogiannis,D.  1853 VanStraten,G.  1607
Tsironi,T.N.  559,1851 Vanaga,I.   2199
Tsotsas,E.   553 Vanin,F.M. 37,227,861,1033,
Tsuta,M. 139,209,455,1057,1281, 1143,1435
1883,2239 Vanoli,M.   895,987,1177,2237
Tubeileh,A.  1371 Vanzo,A.   841
Tucker,G.   1431 Varas,F.   1537
Türköz,G.   851 Varas,J.   1289
Turchiuli,C.   47 Vareltzis,P.K.  1955,1957
Turpin,A.S.   1615 Vargas,M.   41,857,859
Tuta,S.   1755 Varzakas,T.   1103,1923,2147
Tylewicz,U.   515,565,797,1011 VasconcelosPereira,A.  1327,1359
Tyrovouzis,N.A.  1797 Vasheka, O.M.  2023
Tys,J.   1759 Vasilescu,C.  1899
Tzia,C. 943,1021,1291,1345,1649, Vásquez,C.   1075
1775 VásquezͲForttes,F.A.  2091
Tzoumaki,M.V.  129,1055 Vaudagna,S.R.  1655
Uchino,T.   1859 VazͲPires,P.  827
Uemura,K.   73,573,937 Vázquez,M.  1413
Ünalbengör,G.F.  1453 VázquezSolorio,S.C.  1111
Ueno,M.S.   1875,1877 Vega,R.   2161
Ueno,S.   369,1145 VegaͲGalvez,A.  1677,2049
Uhl,C.   1551 Veggi,P.C.   2167,2189,2191
Umeda,M.   9 Velasco,X.   167
Umpuch,C.   385 Velasquez,P.  905

I-22
VelázquezͲCastillo,R.R.  1129,1221 Watanabe,H.  1561
VelézͲRivera,N.  1753 Watanabe,T.  369
Velickova,E.  515 Watanabe,Y.  927
Veljoviđ,M.   2057,2143 Watté, R.  139,2239
Velliou,E.G.  475 Wedekind,J.  1959
Velloso,M.A.C.  1233 Weese,J.   1871
Veloso,C.M.  1097 Weich,H.   877
VenegasͲCubillos,G.  1675 Weisz,G.M.  617
Venema,P.   11,1041 Welderufael,F.T.  395
Ventura,S.P.M.  1703 Welt,B.   93
Veras,A.O.M.  1353,1361 WeltiͲChanes,J.  1671,2045,2139
Verasztó,B.   2213 Werkmeister,R.  757
Verboven,P. 139,843,1057,2227,2231, Werner,F.   949
2239,2245,2249,2251 Wessink,E.   2245,2249,2251
Verdini,R.A.  973,975 Weyandt,A.J.  1595
Vergeldt,F.J.  249 Widell,K.   315
Verhé,R.   1953 Widell,S.   565
Verhoelst,E.  139,2239 Wilke,W.   1481
Verlinden,B.E. 843,1175,1177,2227,2229, Wilkinson,B.  1373
2237,2245,2249 Willems,D.   2225
VernonͲCarter,J.  2097 Williams,J.G.  189,191,1275
Vervoort,L.   351,367 Williams,P.D.  567
Veytsman,B.  889 Wilson,A.   247
Vicente,A.A.  293,413,673,1015,1069 Wilson,D.I.   709,835,1559
Vicente,E.   1875,1877 Wilson,N.   2251
Vicente,S.   1251 Windhab,E.J.  635,643,671,783
VidalͲBrotóns,D.  2089,2091 Windtb,C.   303
Vidigal,M.C.T.R.  1107 Wissink,E.   2241
Vieira,G.S.   1067 Witekb,M.   303
Vignolle,M.  1463 WolfMaciel,M.R.  2063
Vilas,C.   281,1409,1531,1537 Wong,S.Y.   727
Villafin,M.   281 WounTan,L.  309
VillalobosͲCarvajal,R.  113 Wu,S.ͲC.   2125
Villalón,M.   1079 Wunderlich,J.  107,699,1551
Villar,M.A.   399,745 Xanthopoulos,G.  97,1337
Villarroel,M.  829 Xenakis,A.   2127
Villena,P.   1827 Xiao,H.ͲW.   2013
Villeneuve,S.  1261,2033 Xiong,Y.   1469
Viotto,L.A.   551 Xotlanihua,C.E.  1603
Vlahavas,G.  1301 Xu,Z.   439,1031
Vodyanova,I.V.  897 Yahya,F.   329,1511
Voidarou,C.  1013 Yamada,H.   1859
Voigt,T.   731 Yamamoto,C.  7,907
Voilley,A.   373,1919 Yamamoto ,S.  1743
Vollebregt,M.  343 Yaman,D.B.  851
vonStaszewski,M.  833 Yamazaki,H.  15,1051
Vorobiev,E.  403,909,1215,1623,1939 Yang,B.   1983,1985
Voswinkel,L.  381 Yang,S.ͲC.   831
Vraniđ,D.   1285 Yang,S.W.   1035
Vuataz,G.   243,1137 Yanniotis,S. 69,163,341,399,
Wada,Y.   73 1337,1885,2055
Wagner,J.R.  1691 Yano,T.   283,1457
Walker,A.   229 Yavuzdurmaz,H.  2043
Walker,S.   609,1833 Yeganeh,R.   171,1915
Walnum,H.T.  315 Yin,L.ͲJ.   931
Walz,E.   21,121 Yonezawa,Y.  441
Wang,J.   993 Yoo,J.   2073
Wang,J.ͲC.   77 Yoo,S.ͲH.   2073
Wang,L.ͲH.   2013 York,D.W.   709
Wang,P.   931 Yoshii,H.   7,255,907,991
Wang,W.   83 Yoshimoto,N.  1743
Wang,X.   383 Yotsuynagi ,K.  2145
Wang,Y.   795 Youbin,Z.   1673
Warning,A.   1401 Young,A.   247
Wasser,I.   773

11th International Congress on Engineering and Food - Athens, Greece, 2011 I-23
 Youssef,K. 1089,1091,1979,1981
Yuferov,V.   1645
Yunchuan,Z.  1673
Zahedi,A.   1297
Zaidi,S.   81,967,1207
Zanin,G.ɀ.   917,1235
Zaritzky,N.E.  137,743,1395
Zdunek,A.   9,1053,1763
Zelem,M.ͲC.  2251
Zerbe,J.   99
Zhang,L.   91,185,211
Zhang,Y.   439,1617,1659
Zhang,Z.   411,521,711
Zhao,D.   993,1983
Zhao,F.   1985
Zhao,Q.   1985
ZhengRen,F.  1333
Zhenghe,X.   1123
Zhong,Q.   13
Zhou,B.   411
Zhou,L.   439,1617
Zhou,M.   383
Zhou,W.   83,487,647
Zhu,P.   459
Zidová,P.   1695,1707
Zieliŷska,D.  1881
Zieliŷski,H.   1881
Zietz,M.   423
Zikas,A.   1273
Zinoviadou,K.G.  101,879
Zivkoviđ,D.   1285,2083
Zogzas,N.P.  2077,2123
Zoldan,V.C.  953
Zülicke,F.   1633
ZulianiStroppa,V.L.  1293
Zúñiga,R.N.  67,921,935
ZúnigaͲHansenm,M.E.  2161
Zwietering,M.H.  493

I-24
 

InternationalAssociationforEngineeringandFood


LISTOFCOUNTRYDELEGATES


11th International Congress on Engineering and Food - Athens, Greece, 2011 I-25
1.PASTIAEFPRESIDENTS(Retired)

JeanJ.Bimbenet(France)
Ronald.Jowitt(UK)
PekkaLinko(Finland)
MarcLeManguer(Canada)
WalterSpiess(Germany)

2.APPOINTEDDELEGATES

Argentina
 AsociaciʍnArgentinadeTecnʍlogosAlimentarios(AATA)
 StellaM.Alzamoraalzamora@indust.di.fcen.uba.ar

Australia
 AustralianFoodEngineeringAssociation(AFEA)
 MinhNguyenMinh.Nguyen@newcastle.edu.au

Brazil
 SociedadeBrasileiradeCiʃnciaeTecnologiadeAlimentos
 PauloSobralpjsobral@usp.br

Canada
 FoodEngineeringDivisionoftheCanadianInstituteofFoodScienceandTechnology(CIFST)
 MicheleMarcottemarcotte@agr.gc.ca

CzechRepublic
 CzechSocietyofChemicalEngineering
 MilanHouskamilan.houska@vupp.cz

Chile
 InstitutoChilenodeIngenierʆaparalosAlimentos(IChIA)
 JoseM.Aguilerajmaguile@ing.puc.cl

China
 ChineseMechanicalEngineeringSociety(CMES)
 ShujunLilisj@caams.org.cn

France
 AssociationdesChimistes,IngɿnieursetCadresdesIndustriesAgricolesetAlimentaries
(ACIA)
 GillesTrystramTrystram@ensia.inra.fr
 SocieteFrancaisedeGeniedesProcedes,SFGP
 JosephBoudrantjoseph.boudrant@ensaia.inplͲnancy.fr
 


I-26

Germany
 ChemicalEngineeringDivision(GVC)oftheGermanAssociationofEngineers(VDI)
 HelmarSchuberthelmar.schubert@kit.edu

Greece
 TechnicalChamberofGreece(TCG)
 GeorgeD.Saravacosgsaravac@otenet.gr

Ireland
 InstitutionofEngineersofIreland
 DaͲWenSundawen.sun@ucd.ie

Italy
 AssociazioneItalianadiIngegneriaChimica(AIIC)
 MauroMoresimmoresi@unitus.it

Japan
 JapanSocietyforFoodEngineering(JSFE)
 OsatoMiyawakiosato@ishikawaͲpu.ac.jp

Korea
 KoreanSocietyforIndustrialFoodEngineering(KSIFE)
 YegͲHeeChoiyhechoi@knu.ac.kr

Mexico
 InstitutoMexicanodeIngenierosMexicanos(IMIQ)
 JorgeWeltiͲChannesjwelti@mail.udlap.mx

Netherlands
 DutchInstituteofEngineers
 RemkoBoomRemko.Boom@wur.nl

NewZealand
 FoodEngineeringAssociationofNewZealand
 MohammedFaridm.farid@auckland.ac.nz

Russia
AllͲRussianResearchandTechnologyAssociationofDistillationandAlcoholicBeverages
Industries(ARͲRTADABI)
 V.I.Tuzhilkintuzhilkin@mgupp.ru

Singapore
 SingaporeInstituteofFoodScienceandTechnology(SIFST)
 WeibiaoZhouchmzwb@nus.edu.sg


11th International Congress on Engineering and Food - Athens, Greece, 2011 I-27

SouthAfrica
SouthAfricanAssociationofFoodScienceandTechnology(SAAFoST)orSAInstituteof
AgriculturalEngineers(SAIAE)
 AndrewMurrayamurray@pixie.co.za

Spain
 FoodWorkingPartyoftheEuropeanFederationofChemicalEngineering
 PedroFitopfito@tal.upv.es

Sweden
 SwedishSocietyofFoodTechnology(SSFT)
 PeterDejmekPetr.Dejmek@food.lth.se

Thailand
 FoodScienceandTechnologyAssociationofThailnd
 SakamonDevahastinsakamon.dev@kmutt.ac.th

UnitedKingdom
 InstitutionofChemicalEngineers(IChemE)
 PeterFryerp.j.fryer@bham.ac.uk
 InstitutionofMechanicalEngineers
 DonIvesdon@ivesͲconsultants.demon.co.uk
 SocietyforChemicalIndustry(SCI)
 K.Niranjanafsniran@reading.ac.uk

UnitedStates
 AmericanInstitutionofChemicalEngineers(AIChE)
 MartinOkosokos@purdue.edu
 AmericanSocietyofAgriculturalEngineers(ASAE)
 DennisHeldmandrheldman@earthlink.net
 FoodEngineeringDivisionoftheInstituteofFoodTechnologists(IFT)
 R.PaulSinghrpsingh@ucdavis.edu

3.RETIREDDELEGATES

Finland
 EngineeringAssociationofFinland(EAF)
 PekkaLinkoplinko@cc.hut.fi

Portugal
 ColigioNacionaldeEngenhariaQuʆmica(CNEQ)
 AugustoG.Medinaspiporto@spi.pt

I-28
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