Córdoba,

Argentina

VI Iberoamerican Conference on Supercritical Fluids,

Córdoba, Argentina, 2023.
PROSCIBA is a technical conference focused on supercritical fluids fundamentals and applications,
where participants from Iberoamerican countries, as well as from other regions, can share and
exchange their research, expertise, and new ideas, and attend high-standard plenary and keynote
lectures from both academy and industry. Its history goes back to the 1990’s, when the pioneering
Brazilian Meetings on Supercritical Fluids began. From 2007, these meetings were extended to the
Iberoamerican community under the name of PROSCIBA, which is held every three years since then:

2007 – I Prosciba – Iguassu Falls, Argentina and Brazil

2010 – II Prosciba – Natal, Brazil
2013 – III Prosciba – Cartagena, Colombia
2016 – IV Prosciba – Viña del Mar, Chile
2019 – V Prosciba – Campinas, Brazil
2023 - VI Prosciba – Los Cocos, Córdoba, Argentina

VI Iberoamerican Conference on Supercritical Fluids : Prosciba 2023, Córdoba,

Argentina : Book of Abstracts / compiled by Raquel Evangelina Martini, María Laura
Goñi, Alfonsina Esther Andreatta, Nicolás Alberto Gañan. - 1a ed. - Córdoba :
Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales,
2023.

Digital Book, PDF

Digital File: for download and online version

ISBN 978-950-33-1775-4

1. Chemical Engineering. 2. New Technologies. I. Martini, Raquel Evangelina. II.

Goñi María Laura. III. Andreatta, Alfonsina Esther. IV. Gañan, Nicolás Alberto.
Compilers.

CDD 660.071
Organizing Institutions

Sponsors and Support

Local Organizing Committee
Nicolás Gañán (Coordinator) – Universidad Nacional de Córdoba (Argentina)
Raquel Martini – Universidad Nacional de Córdoba (Argentina)
María Laura Goñi – Universidad Nacional de Córdoba (Argentina)
Martín Cismondi – Universidad Nacional de Córdoba (Argentina)
Gerardo Pisoni – Universidad Nacional de Córdoba (Argentina)
Facundo Mattea – Universidad Nacional de Córdoba (Argentina)
Juan Manuel Milanesio – Universidad Nacional de Córdoba (Argentina)
Alexis Velez – Universidad Nacional de Córdoba (Argentina)
José Scilipoti – Universidad Nacional de Córdoba (Argentina)
Lautaro Luján – Universidad Nacional de Córdoba (Argentina)
Noelia Machado – Universidad Nacional de Córdoba (Argentina)
Ramsés Melean – Universidad Nacional de Córdoba (Argentina)
Florencia Muratore – Universidad Nacional de Córdoba (Argentina)
Julieta Quinzio – Universidad Nacional de Córdoba (Argentina)
Andrea Carolina Rodriguez Ruiz – Universidad Nacional de Córdoba (Argentina)
Maricel Santos – Universidad Nacional de Córdoba (Argentina)
Federico Benelli – Universidad Nacional de Córdoba (Argentina)
Janet Chinellato – Universidad Nacional de Córdoba (Argentina)
Paulina Falletti – Universidad Nacional de Córdoba (Argentina)
Pablo Hegel – Universidad Nacional del Sur (Argentina)
Natalia Cotabarren – Universidad Nacional del Sur (Argentina)
Mariana Fortunatti Montoya – Universidad Nacional del Sur (Argentina)
Francisco Sánchez – Universidad Nacional del Sur (Argentina)
Mariana González Prieto – Universidad Nacional del Sur (Argentina)
Aylen Merlo Raimondi – Universidad Nacional del Sur (Argentina)
Alfonsina Andreatta – Universidad Tecnológica Nacional, FR San Francisco (Argentina)

International Organizing Committee

Conference Regional Chairs:
Selva Pereda – Universidad Nacional del Sur (Argentina)
Ignacio Gracia – Universidad de Castilla La Mancha (Spain)
Sandra Regina Salvador Ferreira – Universidade Federal de Santa Catarina (Brazil)
Herminio Sousa – Universidade de Coimbra (Portugal)

María José Cocero – Universidad de Valladolid (Spain)

Lúcio Cardozo-Filho – Universidade Estadual de Maringá (Brazil)
Camila Gambini Pereira – Universidade Federal do Rio Grande do Norte (Brazil)
Gustavo Bolaños – Universidad del Valle (Colombia)
José Manuel Del Valle – Pontificia Universidad Católica de Chile (Chile)
Julián Martínez – Universidade Estadual de Campinas (Brazil)

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International Scientific Committee

María Angela de Almeida Meireles – Universidade Estadual de Campinas (Brazil)

Esteban Brignole – Universidad Nacional del Sur (Argentina)
Erdogan Kiran – Virginia Tech (USA)
Jerry King – University of Arkansas (USA)
Eckhard Weidner – Ruhr Universität-Bochum and Fraunhofer Institute-UMSICHT (Germany)
Cor Peters – Colorado School of Mines (USA)
Nora Ventosa – Institute of Materials Science of Barcelona – CSIC/CIBER-BBN (Spain)
Sagrario Beltran Calvo – Universidad de Burgos (Spain)
Elena Ibañez – Instituto de Investigación en Ciencias de la Alimentación-CSIC (Spain)
Danilo Cantero – Universidad de Valladolid (Spain)
Elisabeth Badens – Aix-Marseille Université (France)
Aaron Scurto – University of Kansas (USA)
Andrea Del Pilar Sanchez Camargo – Universidad de los Andes (Colombia)
Carlos A. García-González – Universidad Santiago de Compostela (Spain)
Feral Temelli – University of Alberta (Canada)
Ignacio Vieitez – Universidad de la República (Uruguay)
Iolanda de Marco – Università degli Studi di Salerno (Italy)
Irena Zizovic – Wroclaw University of Science and Technology (Poland)
Luís Padrela – University of Limerick (Ireland)
Mara Braga – Universidade de Coimbra (Portugal)
Marcus Petermann – Ruhr-Universität Bochum (Germany)
Marleny D. Aranda Saldaña – University of Alberta (Canada)
Owen Catchpole – Callaghan Innovation (New Zealand)
Rita Duarte – Universidade Nova de Lisboa (Portugal)
Sílvio Vieira de Melo – Universidade Federal da Bahia (Brazil)
Željko Knez – Univerza v Mariboru (Slovenia)

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 Plenary Lectures

“Touring the Supercritical Space”.

Esteban A. Brignole
Universidad Nacional del Sur (Argentina)

“The role of sub- and supercritical fluids in the processing of cannabis/hemp”.

Jerry King
Arkansas University (USA)

“Advances on the use of sustainable solvents for pharmaceutical and biomedical applications”.
Ana Rita Duarte
Universidade Nova de Lisboa (Portugal)

“Supercritical Fluid Technology for Food Processing”.

Julián Martínez
UNICAMP (Brazil)

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 Keynotes

“Supercritical fluids technology and sustainability: some opportunities, many challenges”.

Speaker: Gustavo Bolaños (Universidad del Valle, Colombia)

“Hydrothermal extractions-reactions for circular bioeconomy”.

Speaker: Danilo Cantero (Universidad de Valladolid, España)

“Simulation of heat transfer effects on the supercritical CO 2 extraction at a two-vessel industrial

plant”.
Speaker: Felipe Toledo (Pontificia Universidad Católica de Chile)

“Integrated and sustainable pressurized fluid processes for biorefinery of plant and animal by-
products”.
Speaker: Marleny Saldaña (University of Alberta, Canada)

“Important aspects and challenges for the supercritical extraction processes in industry”.
Speaker: Pedro Ivo Nunes (Singularity, Brazil)

“Processing of hemp seed and cannabis flowers - a New Zealand perspective”.

Speaker: Owen Catchpole (Callaghan Innovation, New Zealand)

“Ultra-high pressure processes - Design and examples”.

Speaker: Zeljko Knez (University of Maribor, Slovenia)

“Development of an online integrated method for analyzing bioactive compounds: Pressurized Liquid
Extraction coupled with High-Performance Liquid Chromatography”.
Speaker: Juliane Viganó (Universidade Federal de Sao Carlos, Brazil)

“Come into a new era of purification”.

Speaker: Assia Kone (SFE Process, France)

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“Complex fluid phase behavior in binary and ternary systems, including the critical region”.
Speaker: Cor Peters (Colorado School of Mines, USA)

“ScCO2-impregnation of clove extract in polycarbonate, poly(L-lactic acid) and their blends”.

Speaker: Mathilde Champeau (Universidade Federal ABC, Brazil)

“Selection of green extraction solvents using the Hansen solubility parameters approach”.
Speaker: Andrea Sánchez-Camargo (Universidad de los Andes, Colombia)

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 Oral Communications

008 – “Simultaneous extraction of macauba oil and active compounds from olive leaves using
pressurized propane.” J. M. Jaski, C. E. Barão, C. da Silva, L. Cardozo-Filhoa, and V. F. Cabral.
Speaker: Vladimir Cabral, Universidade Estadual de Maringá

009 – “Development of β-sitosterol-loaded nanocapsules obtained by supercritical fluid extraction of

emulsion process for potential use in functional foods.” D. Cerro, C. Streitt, A. Rojas, J. Romero, and
A. Torres.
Speaker: Daniela Cerro, University of Santiago de Chile

010 – “Concentration of oxygenated terpenes in orange essential orange by supercritical fluid

adsorption.” V. Saldanha Carvalho, J. Viganó, L. H. Fasolin, and J. Martínez.
Speaker: Victor Saldanha Carvalho, UNICAMP

021 – “Supercritical CO2 and melt-extrusion to produce enhanced PLGA scaffolds.” I. Gracia, F.
Carrascosa, J.M. García, M.J. Ramos, J.F. Rodríguez, and M.T. García.
Speaker: Ignacio Gracia, University of Castilla-La Mancha

023 – “High-pressure sequential extractions using aqueous solutions of deep eutectic solvents (DES):
a sustainable process with Green Certificate.” L. Benvenuttia, A. A. Ferreira Zielinski, and S. R.
Salvador Ferreira.
Speaker: Laís Benvenuttia, Federal University of Santa Catarina

034 – “Kappaphycus alvarezzi biorefinery: PLE and SFE applications for phenolics and carragenan
recovery and aerogel drying.” A. R. Rudke, C. J. De Andrade, and S. R. S. Ferreira.
Speaker: Cristiano J. De Andrade, Federal University of Santa Catarina

039 – “Antioxidant extracts from olive pomace by compressed fluid extraction technologies:
comparison between SFE and PLE.” C. Dauber, M. Romero, A. Gámbaro, E. Ibáñez, and I. Vieitez.
Speaker: Cecilia Dauber, Universidad de la República

046 – “Effect of operational variables on the development of PLA/PBAT active foams using
supercritical fluids.” P. Rivera, A. Torres, J. Romero, F. Rodríguez, and M. J. Galotto.
Speaker: Alejandra Torres, University of Santiago de Chile

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048 – “Supercritical CO2 assisted impregnation of pink pepper essential oil in gelatin-siloxane gels
for biomedical applications.” U. Silva do Nascimento, B. Guzzo da Silva, and L. Passos Maia-Obi.
Speaker: Ligia Maia-Obi, Federal University of ABC

050 – “Catalytic synthesis of ethyl levulinate aided by supercritical carbon dioxide.” Authors: M. E.
Zakrzewska, A. B. Paninho, A. R. C. Duarte, and A. V. M. Nunes.
Speaker: Malgorzata Zakrzewska, NOVA School of Science and Technology

051 – “Evaluation of the lipid fraction of tucumã-do-Amazonas (Astrocaryum aculeatum) almond

extracts obtained by SFE.” L. M. S. Carvalho, R. Grimaldi, and J. Martínez.
Speaker: Luciedry Carvalho, UNICAMP

053 – “Supercritical extraction from Citrus reticulata Blanco with carbon dioxide.” M. G. de F. Batista,
M. B. Kolicheski, F. A. Pedersen Voll, and M. L. Corazza.
Speaker: Marcelle Batista, Federal University of Paraná

056 – “Influence of phase behavior on enzymatic esterification of residual oils using propane as
solvent: experimental study and thermodynamic modeling.” B. M. Nogueira, F. W. Tavares, and P. M.
Ndiaye.
Speaker: Bruno Nogueira, Universidade Federal do Rio de Janeiro

057 – “Effect of supercrititical CO2 extraction as pretreatment to obtain c-phycocyanin from spirulina
(Arthrospira maxima).” J. A. López-Limón, M. A. Rios-Corripio, G. Romero-de la Vega, A. S.
Hernández-Cázares, J. V. Hidalgo-Contreras, and R. del A. Mellado-Pumarino.
Speaker: Juana Lopez-Limón, Universidad Iberoamericana Puebla

060 – “Extraction of neutral oil and polyphenolic fractions of spent coffee grounds: single step and
sequential extractions with compressed fluids.” M. Nolasco Araujo, N. do Carmo Diniz, F. Hamerski,
G. Varela Garcia Lesak, J. C. de Carvalho, and M. L. Corazza.
Speaker: Marcos Corazza, Federal University of Para

062 – “Interfacial description of binary systems containing CO2, n-butanol and H2O.” R. Villablanca-
Ahues, R. Nagl, T. Zeiner, and P. Jaeger.
Speaker: Rafael Villablanca – Ahues, Tecnische Univetsität Clausthal

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065 – “Extraction of volatile oils from aromatic Ecuadorian plants.” L. I. Jaramillo, E. Vera, J. Boujila,
and S. Camy.
Speaker: Lorena Jaramillo, Escuela Politécnica Nacional / Université de Toulouse.

069 – “Nutraceutical delivery via SC-CO2: Lentil protein and pectin based aerogels to encapsulate β-
carotene.” S. Mekala and M. D. A. Saldaña.
Speaker: Srujana Mekala, University of Alberta

073 – “Ionic gelation assisted by compressed fluids (IGACF): a new process for microencapsulation of
bioactives in chitosan.” C. Pardo-Castaño and G. Bolaños.
Speaker: Camilo Pardo Castaño, Universidad del Valle

077 – “CO2 + ethanol assisted impregnation of chitosan microparticles with curcumin.” P. H. F.

Tondo, R. M. F. Vargas, and E. Cassel.
Speaker: Eduardo Cassel, Pontifícia Universidade Católica do Rio Grande do Sul

090 – “Comparison of emerging extraction techniques for recovering phenolic compounds from
avocado byproducts using a Natural Deep Eutectic Solvent based on chloride choline and lactic acid.”
J. F. Grisales-Mejía, V. Cedeño-Fierro, J. Pala Ortega, H. G. Torres-Castañed, M. M. Andrade-
Mahecha, H. A. Martínez-Correa, G. Álvarez-Rivera, J. A. Mendiola, A. Cifuentes, and E. Ibañez.
Speaker: Juan F. Grisales-Mejía, Universidad Nacional de Colombia

091 – “Extraction and evaluation of peach (Prunus persica) seed oil obtained by supercritical CO2
extraction.” F.Huayta, H. Pajan, H. Obregón, J. Sedano, J. Paul Rosales, J. Argumé, and J. Palacios.
Speaker: Fredy Huayta, Pontificia Universidad Católica del Perú

108 – “Supercritical CO₂ extraction of oil from soldier fly larva meal (Hermetia illucens L) – yield and
antioxidant activity.” V. A. Cruz and A. L. De Oliveira.
Speaker: Vanessa Cruz, University of São Paulo

110 – “New Deep Eutectic Solvents based on hyperbranched polyglycerols and superbases for CO2
absorption.” G. Silveira dos S., D. Miranda de S. C., and R. C. Bazito.
Speaker: Gabriel Silveira Dos S., Institute of Chemistry University of São Paulo

118 – “Chitosan-TPP based aerogels for tissue engineering applications.”C. S. A. Bento, J. P. Ruivo,
H. C. de Sousa, A. M. A. Días, and M.E. M. Braga.
Speaker: Cristiana Bento, University of Coimbra

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119 – “High pressure packed column for fractionation of low volatile and viscous mixtures.
Conceptual design of glycerol acetates refinement with scCO2.” M. Fortunatti Montoya, F. Sanchez,
P. Hegel, and S. Pereda.
Speaker: Mariana Fortunatti Montoya, PLAPIQUI-UNS-CONICET

121 – “Extraction of bioactive compounds from Scaptotrigona depelis pollen using supercritical
carbon dioxide.” A. Gomes da Silva, E. Gama Ortiz Menezes, and R. Nunes de Carvalho Junior.
Speaker: Adriane Gomes da Silva, Universidade Federal do Pará

125 – “Production of homogeneous biofuel mixtures by supercritical transesterification: experimental

results, process simulation and life cycle assessment.” L. Molina and V. F. Marulanda.
Speaker: Victor Marulanda, Universidad de La Salle

128 – “Supercritical carbon dioxide processing of maritime pine (Pinus pinaster): sorption study.” J.
Leocádio, L. Fonseca, M. E.M. Braga, and H. C. Sousa.
Speaker: João Leocádio, University of Coimbra

135 – “Phase boundaries and copolymerization of acrylic acid + butyl methacrylate + carbon dioxide
under high-pressure single-fluid initiation.” R. S. Meleán Brito, J. E. Tasque, M. C. Strumia, F. Mattea,
J. M. Giussic, and J. M. Milanesio.
Speaker: Juan M. Milanesio, IPQA-UNC-CONICET

143 – “Supercritical CO2-assisted impregnation of PLA films with R-carvone. I: Effect of processing
on thermal and crystallinity properties.” P. P. Miranda, N. A. Gañan, R. E. Martini, and M. L. Goñi.
Speaker: María Laura Goñi, IPQA-UNC-CONICET

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 Poster Presentations

001 – “Phase behavior of ethanolic Passion Fruit (Passiflora edulis sp.) bagasse extract in
supercritical CO2”.
E. J. Santos de Araujo, A. J. Oliveira Braga, P. Matar Ndiaye, and J. Martínez

002 – “Ultrasound-assisted Pressurized Liquid Extraction from Tucumã-do-Amazonas Almonds

(Astrocaryum aculeatum) Defatted with Supercritical CO2”.
M. de P. Kraüss Ferreira, L. M. S. Carvalho, and J. Martínez

003 – “Supercritical CO2 fractionation of passion fruit by-products extract using zeolite 13-X to
concentrate phytosterols and squalene.”
L.C. dos Santos, R. B. Silva, E. Scopel, T. Hatami, C. Rezende, and J. Martínez

004 – “Production of vitamin C-vitamin B3 cocrystals by the gaseous antisolvent (GAS) technique.”
C. A. Balbinot Filho, J. L. Dias, E. A. Rebelatto, S. R. S. Ferreira, and M. Lanza

005 – “Effect of temperature and solvent composition on the pressurized liquid extraction of
phenolic compounds from spent hop.”
A. M. B. Oliveira, J. Viganó, and J. Martínez

006 – “Supercritical impregnation of phenolic compounds of passion fruit (Passiflora edulis sp.)
bagasse extract in corn starch aerogels: effect of operation mode.”
E. J. Santos de Araujo and J. Martínez

007 – “Pressurized liquid extraction of phenolic compounds from pequi almond (Caryocar brasiliense
Camb.).”
I. F. Morenoa and J. Martínez

011 – “Recovery of bioactive compounds from sesame cake by combined high-pressure methods.”
M. A. de Avila Souza, A. R. Rudke, and S. R. Salvador Ferreira

012 – “Optimization of pressurized liquid extraction used to recover flavonoids present in testa shell
from Anacardium Occidentale.”
L. J. da Silva, E. Sousa de Brito, and S. R. Salvador Ferreira

014 – “Supercritical CO2 extraction of essential oil from guarana (Paullinia cupana) peel.”
R. P. F. de Melo, R.C. Moreira, J. L. Bicas, N. C. Bastos, A. L. Atroch, T. P. De Souza and J. Martínez

015 – “Fractionation of α-terpineol and limonene by supercritical fluid adsorption.”

R. P. F. de Melo, B. B. Kienast, L. C. dos Santos, R. C. Moreira, J. L. Bicas, and J. Martínez

016 – “Supercritical fluid extraction of alkaloids from Cestrum plants.”

F. D. Sanches and J. Martínez

017 – “Effects of temperature and molar ratio between acetic anhydride and α-terpineol on terpinyl
acetate production in SC-CO2.”
R. C. Moreira, R. P. F. de Melo, J. Martínez , and J. L. Bicas

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018 – “Recovery of phenolic compounds of the Melão-de-São-Caetano (M. charantia) peel by high-
pressure extraction methods.”
M. C. de Sousa, A. R. Rudke, and S. R. S. Ferreira

019 – “Chlorella vulgaris deconstruction by green sequential extractions: the potential of

(bio)surfactant modifiers.”
G. G. González, S. R. Salvador, and C. J. De Andrade

020 – “Influence of chemical composition on the properties and foamability with supercritical CO2
in thermoplastic polythiourethanes.”
I. Gracia, P. Belmonte, J. M. García-Vargas, J. F. Rodríguez, and M. J. Ramos

022 – “Assessment of pressurized solvents extractions from grape pomace residues.”

V. S. Carvalho, L. C. Freitas, and M. L. Corazza

024 – “Supercritical fluid technology as a tool to valorize bacuri fruit (Platonia insignis Mart.) shell.”
L. Cantão Freitasa, F. C. Seabra Pires, T. Costa de Araújo, and R. Nunes de Carvalho Junior

025 – “Pressurized propane extraction of umari (Poraqueiba sericea Tul.) oil: extraction kinetics and
fatty acid composition.”
L. Cantão Freitasa, V. Souza Carvalho, M. L. Corazza, and M. L. Massona

027 – “High Pressure Liquid Impregnation of Barbatimão Stryphnodendron adstringens (Mart.)

Coville bark extracts in biopolymeric membranes.”
P. C. Veggi, L. F. Campos, L. E. do Carmo, J. Viganó, L. P. Mazur, and L. H. I. Mei

028 – “Pressurized liquid extractions of proteins and biocompounds with antioxidant activity from
banana inflorescence (Musa acuminata cv. Cavendish).”
G. E. Motta, G. A. Valencia, and S. R. Salvador Ferreira

029 – “A model sugar cane waste biorefinery based on sub and supercritical water processing: a
LCA case study.”
V. F. Marulanda

030 – “Gurguéia nut (Dipteryx lacunifera Ducke) cake as a source for obtaining extracts rich in
bioactives using pressurized liquid extraction.”
G. Polmanna, P. H. Santos, R. Alves Morais, S. R. Salvador Ferreira, and J. M. Block

031 – “Scaling up of the Extraction of Dill Essential Oil.”

M. Franco and M. D.A. Saldaña

032 – “Pressurized liquid extraction as a promise method to obtain protein from broken black bean
cotyledon.”
R. Fialho Teixeira, L. Benvenutti, D. de Oliveira, and A. A. Ferreira Zielinski

033 – “Supercritical fluid extraction of oil from Mauritia flexuosa pulp: a comparison with traditional
extraction techniques.”
M. P. Carrillo-Bautista, L. L. Orduz-Díaz, J. E. C. Cardona-Jaramillo, and G. Bolaños

035 – “Peach seed cake (Prunus persica): a comparative study of different extraction methods for
the recovery of protein fraction.”
C. R. M. Rudke, T. M. S. Torres, A. A. F. Zielinski, and S. R. S. Ferreira

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036 – “Recovering bioactive compounds from pineapple waste: a comparison between pressurizes
liquid and supercritical fluid extractions.”
F. A. Maia and L. H. Fasolin

037 – “Integration of alternative solvents for enhanced stability of dill (Anethum graveolens L.)
aroma volatile compounds.”
J. Vladic, S. Jokic, K. Aladic, I. Jerkovic, and A. R. Duarte

038 – “Naringin processed by gas antisolvent technique.”

P. V. Oliveira, J. L. Dias, G.P. S. Aguiar, K. Z. Kuhn, O. Sanaiotto, A. C. Provinelli, C. F. Daniel, A. Bortoluzzi, A.
M. Siebel, L. A. Lerin, C. Trapella, L. G. Müller, and J. V. Oliveira

040 – “Recovery of phenolic compounds from pink pepper fruits.”

E. A. Rebelatto, L. Vitali, A. R. Rudke, K. S. Andrade, and S. R. S. Ferreira

041 – “Sequential extraction of tocols and carotenoids from Butia capitata fruits using supercritical
carbon dioxide and pressurized fluids.”
G. S. Lourenço Pereira, P. Tonon de Souza, S. Fraga, and K. Araujo Sampaio

043 – “Recovery of extracts rich in phenolic compounds from buritirana pulp (Mauritella armata)
using high-pressure extraction processes.”
R. Alves Morais, E. A. Rebelatto, S. R. Salvador Ferreira, and J. M. Block

044 – “Production of vitamin C-vitamin B3 cocrystals by the gaseous antisolvent (GAS) technique.”
C. A. Balbinot Filho, J. L. Dias, E. A. Rebelatto, S. R. S. Ferreira, and M. Lanza

045 – “High-pressure phase equilibrium data for carbon dioxide + 2-methyltetrahydrofuran-3-one

and carbon dioxide + 2-methyltetrahydrofuran-3-one + glucose systems.”
E. L. S. Barros, E. A. Rebelatto, D. A. Mayer, and J. V. Oliveira

047 – “Obtaining and characterization of quinoa starch (Chenopodium quinoa Willd.) defatted by
supercritical fluids for its application as biopolymer.”
Y. Aliaga and L. Olivera-Montenegro

049 – “Gelatin-Siloxane xerogel and criogel loaded with ibuprofen by supercritical CO2 assisted
impregnation.”
U. Silva do Nascimento and L. Passos Maia-Obi

052 – “Sequential supercritical extraction of extract of L. rivularis stems using CO2 and ethanol-
modified CO2”.
C. Marillán and E. Uquiche

054 – “Enzymatic copolymerization of ω-pentadecalactone and globalide in supercritical carbon

dioxide.”
R. D. Santos, E. A. Rebelatto, D. A. Mayer, and J. V. de Oliveira

055 – “Supercritical fluid extraction of coffee oils for sunscreen formulations.”

N. Trullo, I. Mejía and G. Bolaños

058 – “Gelatin-Siloxane aerogels by supercritical drying.”

B. Andrade de Campos and L. Passos Maia-Obi

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059 – “Value-added products from wheat straw using pressurized water + ethanol mixtures.”
A.-X. Vidrio-Sahagun and M. D.A. Saldaña

061 – “Multiphase equilibria modeling of CO2 the homologous series of carboxylic acids.”
J. M. Ortega Bacicheti, V. Ferreira Cabral, and F. A. Sánchez

063 – “Techniques to solubilize and adsorb acid dyes in alpaca (vicugna pacos) fiber using
supercritical fluid.”
B. G. Orcón Basilio, D. R. Verástegui Cholan, M. E. Tito Silva, F. V. Huayta Socantaype, J. L. Sedano Malpartida,
R. Chavez, S. M. Figueroa Flores, S. M. Trujillo Navarro, and E. V. Carranza López

064 – “Operation parameters of dyeing alpaca fiber with supercritical fluid.”

B. G. Orcón Basilio, D. R. Verástegui Cholan, M. E. Tito Silva, F. V. Huayta Socantaype, J. L. Sedano Malpartida,
S. M. Figueroa Flores, S. M. Trujillo Navarro, and E. V. Carranza López

066 – “An integrated high-pressure and supercritical CO2-based process for sterilization of alginate-
gelatin aerogels.”
C. S. A. Bento, S. Alarico, N. Empadinhas, H. C. de Sousa, and M. E. M. Braga

067 – “Development of Simulator: Supercritical Extraction Process.”

O. A. F. Oliveira, R. M. F. Vargas, and E. Cassel

068 – “Bioactive Compounds from Yerba-Mate: Supercritical Decaffeination and Polyphenol

Extraction.”
B. Sgarioni, L. C. Angeli, R. M. F. Vargas, and E. Cassel

070 – “Effect of Casearia sylvestris supercritical extracts on levels of serum cholesterol, triglycerides
and glucose.”
P. Benelli, R. C. Pedros, and S. R. Salvador Ferreira

071 – “Processing of rosehip and grapes yields for recovery of biologically active components.”
M. Knez Marevci and Ž. Knez

072 – “Valorization of grape marc (Vitis labrusca) using supercritical extraction: Lab-scale DOE and
pilot-scale validation.”
C. Pardo-Castaño, E. Sanchez, and W. Vallejo

074 – “Encapsulation of niacin in PVP-K30 by gas antisolvent technique.”

M. Silveira, P. V. Oliveira, E. A. Rebelatto, P. H. H. Araújo, C. E. M. Campos, and J. V. Oliveira

078 – “Calculation of critical endpoints and phase diagrams of highly asymmetrical binary
mixtures.”
F. A. Sánchez and M. Cismondi

079 – “Coumarins Supercritical Extraction from Citrus Peel.”

J. J. Mora, H. M. Tavares, E. Cassel, and R. M.F. Vargas

15
081 – “Optimization SFE process of bioactive compounds from Myroxylon balsamum (Estoraque)
seeds.”
L. O. Chañi-Paucar, F. de A. Maia, G. R. Lobato-Calderon, J. Martínez, and L. H. Fasolin

084 – “Efficient flash computation of continuous sets of solid-liquid-vapor equilibria directly related
to laboratory data obtained through the synthetic method.”
M. J. Molina, J. V. Mattos, S. B. Rodriguez-Reartes, L. Cardozo Filho, and M. S. Zabaloy

085 – “Upgrading of biolubricants by CO2 technology.”

A. Pizzano, S. B. Rodriguez Reartes, and P. Hegel

086 – “High-pressure viscosity measurements of orange oil saturated with CO2.”

M. Fortunatti-Montoya, P. E. Hegel, and S. Pereda

087 – “Measurement and correlation of PVT data for the CO2+hydrogen+methanol+water system.”
N. S. Cotabarren, P. E. Hegel, F. A. Sanchez, and S. Pereda

092 – “Hibiscus sabdariffa L. leaves as an alternative source of bioactive compounds obtained by

supercritical technology.”
G. N. Clímaco, R. Vardanega, and L. H. Fasolin

094 – “Solubility of aspirin, ketoprofen and R-(-)-carvone in supercritical CO2 in binary, ternary and
quaternary systems: Effect of co-solutes.”
K. Dujarric, I. T. Coutinho, G. T. Mantuaneli, T. Tassaing, and M. Champeau

098 – “Supercritical CO2 extraction of the compounds of the old cinnamon leaves (Miconia albicans)
and antioxidant activity evaluation.”
V. A. Rambo Cerioli, E. Jair Klein, G. Caroline Peiter, E. A. da Silva, P. A. Palsikowski, V. Ferreira Cabral, and L.
Cardo Filho
099 – “A circular economy approach for the processing of fisheries waste: Pressure-intensified
extraction of valuable products from fish roes.”
L. I. Granone, R. I. Naveira, D. L. Lamas, M. A. Fanovich, M. S. Churio, and Á. E. Massa

100 – “Use of a pressurized aqueous solution of a natural deep eutectic solvent to extract bioactive
compounds from the Hass avocado epicarp.”
J. F. Grisales-Mejía, M. M. Andrade-Mahecha, H. A. Martínez-Correa, J. A. Mendiola, A. Cifuentes, and E.
Ibañez

101 – “Predictive correlation for high-pressure phase equilibria of nitrogen + n-alkane binary
mixtures with the RKPR EoS.”
M. J. Quinzio, F. E. Benelli, S. B. Rodriguez-Reartes, and M. Cismondi Duarte

102 – “Effect of cosolvent (ethanol/water) on the supercritical CO2 extraction of flavonoids from
hydroethanolic suspensions of calafate fruit.”
C. Moreno, G. A. Núñez, L. Cuellar, and J. M. del Valle

103 – “Algorithm for calculating phase diagrams at set initial global composition involving fluid and
solid-solution phases for multicomponent chemically reactive systems.”
M. J. Molina, A. F. Porras-Giraldo, A. Pizzano, S. B. Rodriguez-Reartes, and M. S. Zabaloy

104 – “Determination of process conditions to obtain Macaúba (Acrocomia aculeata) oil using
Supercritical CO2 and its potential for application in new products.”

16
G. C. M. Aires, Kelly R. P. Pantoja, J. M. L. Pinheiro, V. M. S. Bezerra, M. E. F. de Carvalho, E. G. O. Menezes,
and R. N. de Carvalho Junior

105 – “Subcritical fluid extraction of phenolic compounds: an alternative to add value to pecan
nutshells.”
A. C. Rodríguez-Ruiz, M. E. Cardona Jimenez, A. R. Velez, L. M. Agudelo-Laverde, P. R. Santagapita, and R. M.
Bodoira

106 – “Supercritical fluid extraction of antioxidants from "Lorena" avocado seeds.”

J. C. de la Cruz, L. Salazar, I. Mejía, and G. Bolaños

107 – “Supercritical fluid extraction from bark of mururé (Brosimum acutifolium): Sequential
extraction study, Chemical composition, total phenolic compounds and antioxidant activity.”
K. R. P. Pantoja, F. de Freitas M. de Azevedo, E. Gama O. Menezes, H. L. de Araujo, W. Braamcamp de S.
Pinheiro, and R. N. de Carvalho Júnior

109 – “Influence of bed filling on the kinetics of the extraction of phenolic compounds from araçauna
(Psidium myrtoides) using pressurized liquids”.
Y. M. Monroy, S. Fraga, P. Berilli, M. R. Maróstica Jr., M. Villa Nova Rodrigues, and K. Sampaio

111 – “Development of Deep Eutectic Solvents based on amino acids for CO 2 absorption.”
D. Miranda de S. C., G. S. Santos, and R. C. Bazito

112 – “Compositions of fatty acids and tocopherols from macauba pulp (Acrocomia aculeata)
extracted with supercritical fluid.”
Y. M. Monroy, G. S. L. Pereira, C. A. Colombo, and K. A. Sampaio

113 – “Supercritical technology applied to obtain tucumã oil (Astrocaryum vulgare): Kinetic
parameters and chemical composition.”
R. M. Morais, E. G. O. Menezes, N. J. N. da Silva, M. C. R. Ferreira, K. R. P. de Sousa, F. de F. M. de Azevedo,
C. B. G. Teles, and R. N. de Carvalho Júnior

114 – “Physical-chemical studies of araçauna flour (Psidium myrtoides) and extraction yield
obtained by sequential extraction in fixed bed with SFE and PLE technology.”
S. Fraga, Y. M. Monroy, P. Berilli, M. R. Maróstica Jr., D. Gonçalves, and K. Araujo Sampaio

115 – “Determination of process parameters and antioxidant potential of Calycolpus goetheanus

(O. Berg) essential oil obtained by supercritical CO2 extraction.”
R. V. Mota, N. J. N. da Silva, E. G. O. Menezes, M. E. F. de Carvalho, J. M. L. Pinheiro, and R. N. de Carvalho
Junior

116 – “Supercritical extraction of butter from agroindustrial cocoa residue from the amazon.”
J. M. L. Pinheiro, A. P. S. e Silva, K. R. P. Pantoja, R. Vitti Mota, G. I. de A. Campos, G. C. M. Aires, V. M. S.
Bezerra, M. E. F. de Carvalho, and R.N. de Carvalho Junior

117 – “Extraction of yerba mate (Ilex Paraguariensis) using near-critical CO2 + hydrated ethanol.”
L. I. Granone, F. A. Sánchez, P. E. Hegel, and S. Pereda

120 – “Application of green technologies for the valorization of banana peel from the chips industry:
a biorefinery approach.”
A. del Pilar Sánchez-Camargo, P. Henrique Santos, L. Benvenutti, E. Rojas-Orduña, and S. R. Salvador Ferreira

122 – “Theoretical Study of the Fractionation of Glycerol Laurates Using SC-CO2.”

17
F. A. Sánchez, N. S. Cotabarren, P. E. Hegel, and S. Pereda

123 – “Kinetics of carotenoids extraction from peels of peach palm fruits (Bactris gasipaes) by CO2
supercritical.”
S. E. G. Souza, E. G. O. Menezes, R. C. Chisté, and R. N. C. Junior

124 – “First Contact Minimum Miscibility Pressure for Enhanced Oil Recovery: Tracing Pressure-
Composition (P-x) Phase Envelopes and Analyzing Injection Fluid Effects.”
F. E. Benelli, G. O. Pisoni, and M. Cismondi

126 – “Assembly of supercritical extractor to operate in semi-continuous mode without CO2

liquefaction.”
A. P. S. Silva, M. C. R. Ferreira, L. M. M. Siqueira, L. B. Santos, L. L. T. Ribeiro, and R. N. Carvalho Jr.

127 – “Supercritical CO2 extracts of ginger (Zingiber officinale R.): kinetic global yield, polyphenol
content, antioxidant activity and its application as antioxidant in olive oil.”
J. Meza-Coaquira and F. P. Cárdenas-Toro

131 – “Unlocking the potential applications of Ganoderma applanatum through ultrasound and
supercritical fluid extraction: A presentation of the state of the art.”
P. C. Torres-Mayanga, J. Vigano, L. O. Chañi-Paucar, S. R. Tovar-Barrientos, C. F. Epiquien-Zorrilla, J. L.
Pasquel-Reátegui, L. Condezo-Hoyos, and M. Rostagno

132 – “Comparison of modified HPAM using hyperbranched brush macromonomer in supercritical

carbon dioxide and controlled polymerization.”
R. S. Meleán Brito, A. Iborra, J. E. Tasque, M. C. Strumia, F. Mattea, I. N. Vega, J. M. Giussi, and J. M. Milanesio

133 – “Supercritical CO2 + co-solvent extraction of essential oil from Mentha piperita and scale up.”
M. D. A. Saldaña and Y. Menga

134 – “Exploring the physicochemical and rheological properties of acrylic acid - co - 2-acrylamido-
2-methylpropane sulfonic acid copolymers synthesized in supercritical carbon dioxide.”
R. S. Meleán Brito, F. Mattea, J. Padró, M. C. Strumia, S. Camy, M. Destarace, and J. M. Milanesio

136 – “Exploring the potential of supercritical carbon dioxide for eugenol impregnation in 3D printed
polylactic acid structures.”
J. Chinellato, M. R. Romero, and F. Mattea

137 – “Delipidation of porcine femur bone and porcine pericardial membrane using supercritical
CO2 extraction.”
J. Reynoso, N. A. Gañan, Alexis Velez, M. Murature, and J. M. Milanesio

138 – “Supercritical CO2 pretreatment and subcritical water extraction of sulfated flavonoids from
Flaveria bidentis leaves.”
P. Falletti, M. F. Barrera Vázquez, P.H. Santos, L.G. Gonçalves Rodrigues, M. Lanza, R. E. Martini, and L. R.
Comini

139 – “Esterification of oleic acid using subcritical isoamyl alcohol.”

M. Santos, C. Rodriguez Ruiz, J. Milanesio, I. Magario, and A. Velez

140 – “Validation of the RK-PR equation of state for the calculation of volumetric properties for
hydroethanolic mixtures at subcritical conditions.”

18
A. C. Rodríguez-Ruiz, G. O. Pisoni, and A. R. Velez

141 – “Supercritical CO2 sorption and diffusion in polylactic acid films: experimental study and
modeling.”
M. L. Goñi, J. Scilipoti, and N. A. Gañán

142 – “Preparation of cellulose aerogels using supercritical drying: Influence of process variables on
aerogel porosity.”
N. D. Machado, V. Pigino, J. Chiavassa, M. L. Goñi, and N. A. Gañán

144 – “Supercritical CO2-assisted impregnation of PLA films with R-carvone. II: Effect of process
variables on mechanical behavior.”
F. Muratore, P. P. Miranda, L. Luján, N. A. Gañán, M. L. Goñi, and R. E. Martini

19
PLENARY LECTURES

20
 Touring the supercritical space

Esteban Brignole

Planta Piloto de Ingeniería Química (PLAPIQUI-UNS-CONICET).

Universidad Nacional del Sur, Bahía Blanca, Argentina.
eabrignole@gmail.com

ABSTRACT

In 1981, Aage Fredenslund presented his groundbreaking Unifac group contribution model at
PLAPIQUI, describing a liquid mixture as a complex interplay of molecular groups. On that
important day a single question from the audience gave birth to Computer Aided Molecular
Design (CAMD). This new journey began with the application of CAMD to search for viable
liquid solvents for the extraction and dehydration of alcohols. To our initial surprise, the results
were inconclusive. However, during my sabbatical leave at the Kemiteknik Institute in
Denmark, a process centered around near-critical propane emerged. This process was based on
the non-classic supercritical properties of propane and its application in the extraction and
dehydration of ethanol, employing the GCEOS model. Our work was presented at the 1991
Boston Supercritical Conference, where we showed the experimental validation of this
innovative process. At the same conference, a dedicated team from Mitsubishi Company
confirmed a pilot plant-scale implementation of all stages of the hot propane process, giving
us an epiphanic moment. Subsequently, we enriched the GCEOS model with a group
contribution with association term based in Whertein theory, thus giving birth to the GCA EOS.
Susana Bottini further expanded our horizons by using propane to investigate the impact of
molecular size asymmetry on the phase behavior of vegetable oils with light hydrocarbons.
This work marked our initial contact with Cor Peters as early as 1997. Both projects demanded
the development of thermodynamic modeling and experimental units, work that was carried
out on both sides of the Atlantic. We entered the world of supercritical reactors when, working
in Delft, Susana obtained the first reliable data on hydrogen solubility in vegetable oils. Just as
remarkable were Selva Pereda's contributions. Her PhD thesis focused on the design of the
phase conditions and solvent selection needed to achieve a homogeneous phase in the
hydrogenation of vegetable oils, defining the field of Phase Equilibrium Engineering. Pablo
Hegel et al applied the principles of phase design to extract natural oils using propane and
carbon dioxide mixtures, under conditions that offered both high solvent power and safety.
This group also ventured into biodiesel production through supercritical transesterification with
methanol. It also delved into measuring densities and phase boundaries using isochoric cells to
evaluate continuous transesterification reactor data.
And finally, throughout this productive collaborative journey, with coleagues from Cordoba
we developed experimental units and thermodynamic models like GCA EOS and RKPR EOS,
along with software tools like GPEC, that further enriched the field of process thermodynamics.

Keywords: Thermodynamic modeling, GCEOS, GCA EOS, Phase Equilibrium Engineering

21
 The Role of Sub-and Super-critical Fluids in the Processing of
Cannabis/Hemp
Jerry W. King*

CFS (Critical Fluid Symposia), Fayetteville, AR 72704, USA

*kingjw100@hotmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Sub- and super-critical fluids have played a significant role in the development of the
cannabis/hemp processing platform, resulting in extracts used in the formulation of a variety
of end-products. Aside from sub- and super-critical carbon dioxide - propane/butane, R134a,
dimethyl ether (DME), and even water, have been used for extraction of the constituents of
cannabis/hemp (cannabinoids, terpenes, etc.). Optimization of these various modes of
extraction is challenging due to the lack of experimentally-determined physicochemical
property data, however EOSs (Equations of State), HSP (Hansen Solubility Parameter Theory)
and application of a molecular dynamics’ method have improved the current situation. The
pros/cons of using the sub-, near-, and super-critical state of the above fluids - with respect to
their selectivity towards the constituents of cannabis/hemp, will be described. As illustrated in
the above graphical abstract, the extraction step is proceeded by selection of the right
cannabis/hemp cultivars (trim/flower), a comminution option, and if desired- decarboxylation
of the cannabis biomass. Optimizing these pre-extraction factors influence the outcome of the

22
 sub- or super-critical extraction as well as the desired end result (product). The morphology
of the cannabis/hemp plant is complex: pretreatment of cannabinoid-terpene trichomes via
sifting and dry-ice or ice water comminution is a common practice among “artisan” processors.
Decarboxylation of the cannabinoid moieties to their neutral forms if desired can be performed
batch-wise in ovens or continuous mode, employing inert gas or vacuo conditions to prevent
cannabinoid oxidation and/or terpene loss from the plant matrix. The occurrence of both acidic
and neutral cannabinoids with the cannabis/hemp material can result in differential rates of
extraction as well as formation of “derivative” cannabinoids such as CBN, CBG, Δ8THC, etc.
– desired or not. With respect to cannabis extraction using the sub--critical fluid state, the
fluids n-propane/-butane, R134a (HFC134), and dimethyl ether are utilized at low extraction
temperatures (<Tc and <T bp) to extract and preserve the terpene and cannabinoids. The
reported conditions are obviously quite different from using these fluids as supercritical media,
although their use under subcritical conditions is somewhat empirical in practice. HFC-134
and DME are touted as better substitutes for propane/butane for producing an alternative to
butane hash oil (BHO), i.e., DME hash oil (DHO). Use of these fluids could be improved by
better extractor temperature control and applying thermodynamic principles with respect to the
chosen pressure/temperature with respect to the V-L locus. The use of CO2-based extraction
represents ~52% of all of the reported cannabis extraction techniques and is highly preferred
in the industry due to its environmentally-benign nature and consumer approval in many
formulated products from these extracts. The “classic” technique is to first perform a CO2
extraction under sub-critical conditions and collect the terpene-enriched fraction, followed by
supercritical CO2 extraction between 10-30 MPa/40-60o C for the cannabinoids, and then
composting the collected fractions for post-extraction refinement, or offered as a “full
spectrum” extract. Neat CO2 extractions up to 100 MPa/50o C have been reported for optimal
extraction of Δ9THC, conditions predicted by HSP. Cannabinoid extraction discrimination can
also be affected under L-CO2 conditions (20-25 C/< Tc) with further enrichment of the neutral
cannabinoids and removal of unwanted co-extractives using in-line sorbent beds, a method that
is also used in subcritical propane/butane extraction technology. Finally, water in the
subcritical state has been applied by this researcher and others for the extraction of
cannabis/hemp biomass usually between 100 – 160O C and requisite pressures (< 2.5 MPa)
required by the V-L equilibria curve. Such conditions might seem odd based on solubility
considerations, but extraction under these conditions allow for in-situ decarboxylation, phase
separation of cannabinoid-rich oil phase from the aqueous medium after pressure reduction,
and potentially, the formation of “desired” cannabinoid derivatives. In conclusion,
cannabis/hemp extraction continues to be an evolving field of activity; several references of
the current state of the art from Espana 1) and (3) Latin America (3) are given below:
(1) Christina Lopez-Olmos et al, Frontiers in Nat. Prods. (2022),10:3389, pp. 1-39.
(2) Masoumeh Pourseyed Lazarjani et al, J. Cannabis Res. (2021) 3:32, pp. 1-15.
(3) Angela Suarez-Jacobo et al, Molecules, (2023) 28:2895, pp.1-15.

Keywords: cannabis, CO2-extraction, decarboxylation, full-spectrum extract, hemp

23
 Advances on the use of sustainable solvents for pharmaceutical
and biomedical applications
Ana Rita C. Duarte*

LAQV/Requimte, Department of Chemistry, NOVA School of Science and Technology, FCT NOVA,
Campus da Caparica, 2829-516 Caparica, Portugal.
*corresponding author email address: aduarte@fct.unl.pt

ABSTRACT
The concept of green solvents was introduced by John Warner and Paul Anastas, in 1998, when
they listed the 12 principles of green chemistry. One of these principles assumes that the use
of solvents, should be as an auxiliary substances, in the synthesis and preparation processes,
and emphasises that they should be made unnecessary wherever possible, and if used they
should be innocuous.[1] The sustainability of a process or a product is, hence, a result of the
complex interaction between the product or process implemented and environment, technology
and economy factors.[2] In the current scenario of a worldwide quest for sustainable and
“greener” alternatives for current applications, in the past two decades the number of reports
on NADES/DES applicability in various situations has rapidly grown. The tailor-made
versatility of these eutectics, meaning in practical terms a wide range of chemical and physical
properties, has resulted in promising studies applied in industry, biotechnology, cosmetics,
pharmaceuticals, food, and health.[3]
In life science applications, several challenges arise. In particular, the formulation of
therapeutic deep eutectic systems (THEDES) revealed to be very promising to enhance the
characteristics of existing drugs and optimize new formulations that could be easily
administrated and more effective, providing higher dissolution rates of the API, enhancement
of solubility, permeability and absorption through tissues, or they can be incorporated in
suitable polymer to potentiate the efficiency of biomedical devices. Coupled with supercritical
fluids it is possible to combine two state of the art green technologies towards the development
of enhanced materials, namely 3D scaffolds, impregnated membranes and particles for drug
delivery. In this communication, these different examples will be presented and discussed.

Keywords: Life sciences; Pharmaceutics; Deep eutectic solvents; Therapeutic deep eutectic
solvents; Green chemistry; Cryopreservation

References:
[1] Anastas P, Eghbali N. Green Chemistry : Principles and Practice. 2010;301–12.
[2] Sheldon RA. The: E factor 25 years on: The rise of green chemistry and sustainability.
Green Chem. 2017;19:18–43.
[3] Paiva A, Craveiro R, Aroso I, Martins M, Reis RL, Duarte ARC. Natural deep eutectic
solvents - Solvents for the 21st century. ACS Sustain Chem Eng. 2014;2:1063–71.

24
 Supercritical fluid technology for food processing
Julian Martinez*

Universidade Estadual de Campinas.

*corresponding author email address: julian@unicamp.br

ABSTRACT
Supercritical fluid technologies have overcome the research boundaries and gained industrial
status in the past decades, due to the unique and attractive features of supercritical fluids that
are well known in our community. The versatility and tunability of supercritical fluid properties
made them excellent solvents for separation processes such as extraction, which stands out as
the main unit operation involving these fluids in research and industrial scales. Moreover, the
use of carbon dioxide as supercritical solvents offers additional advantages, since its mild
critical temperature is adequate to thermally sensible compounds, and it is a safe solvent for
application in products for human consumption, such as food. Given this background, this talk
will offer an overview of supercritical fluids in the food field, mentioning consolidated
industrial processing and some promising applications. As the main application in this area,
supercritical fluid extraction is addressed with a special emphasis on the recovery of food by-
products, but also as a strategy to add value to our biodiversity. Other classical unit operations
that are feasible with supercritical fluids are exposed, such as fractionation, adsorption,
impregnation, particle formulation and chemical reactions, which can be integrated in a
biorefinery approach to provide the complete recovery of a given raw material. Finally, the
current challenges of food industries are presented, such as the demand for natural ingredients,
healthy products, plant proteins and waste recovery. In this context we identify strategies for
supercritical fluids to help overcoming them and offer alternatives for the safe and sustainable
food production in the future.

25
KEYNOTES

26
 Supercritical fluids technology and sustainability:
some opportunities, many challenges

Gustavo Bolaños

Laboratory of Applied Thermodynamics and Supercritical Fluids

School of Chemical Engineering, Universidad del Valle
CL 13 100-00 Edificio E30, Cali, Colombia
gustavo.bolanos@correounivalle.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
The world is attending to an unprecedented revolution regarding the way we transform and use
energy, improve the environmental performance of productive processes, and decrease the
carbon fingerprint of human activities. Governments around the world have been making
efforts to identify and quantify the impact of each country to the problem of the continuous
increase of atmospheric CO2 concentration, and many have detailed plans to achieve low or
net-zero carbon emissions by 2050. Concepts such as “sustainable development” and “circular
economy” have been part of the political discussion in many countries for already a long time.
This revolution presents many opportunities and challenges for researchers from many
scientific and technological fields, including the field of supercritical fluids.
Successful technological solutions, however, require not only scientific understanding and
technological insight (e.g., invention), but also important elements related to
commercialization. Society demands innovation, which some people have expressed with the
equation:
Innovation = invention x commercialization
Note that in this expression the operator is “x” and not “+” as is usually considered by many
scientists and engineers. This expression in many cases challenge the way we researchers
perceive our work.

27
In this presentation, three lines of work for developing sustainable processes that were
performed in our laboratory in a timespan of 25 years will be described, taking into
consideration the technical opportunities involved and, most importantly, some of the
challenges we experienced in the search for innovation. The first work deals with the
preparation of building and encapsulating materials based on silica and lime agglomerates, in
which control of the porosity and other important properties of the material can be
accomplished by infiltration with supercritical CO2. The second work deals with the use of
subcritical water to produce chitin from Aspergillus niger mycelium (a filamentous mold that
is a byproduct of the industrial production of citric acid) and exoskeletons of crustaceans. The
third work deals with the destruction of dielectric oils contaminated with polychlorinated
biphenyls, which, according to international treaties, must be eliminated in an environmentally
friendly way before 2028. These three lines of work were developed to different degrees in its
way to commercialization (the so-called “Technology Readiness Level” or TRL) and allowed
us to identify important non-technical barriers that will be summarized in this contribution.
Some of the barriers are related to differences in culture and even language of academia and
industry; in particular, how time and money are valued. There are other practical barriers,
related to the usual profile of a scientific researcher, and to the structure of administrative
offices created by universities to promote transference of research results to industry.
Overcoming such barriers is essential for the supercritical fluids technology to contribute to
the sustainable development of our nations, and to be considered by society and governments
as a useful field that deserves support.

Keywords: Supercritical fluids, commercialization, materials, PCB, supercritical water, TRL.

28
 Hydrothermal Extractions-Reactions for circular bioeconomy
Danilo Cantero

The Institute of Bioeconomy

*Danilo.Cantero@uva.es

GRAPHICAL ABSTRACT

ABSTRACT
The Institute of Bioeconomy has evolved its supercritical water hydrolysis equipment to a next
level of automatization and control. Our current equipment can perform hydrolysis under
supercritical water conditions with biomass flow of 1 to 5 dry kg/h. This equipment allows us
to quickly evaluate different materials as potential feedstocks. Our main strategy is the use of
inexpensive (like industrial or agricultural residues) as feedstocks. These materials do not
represent a costly raw material. Going deeper in that strategy, we focus in the “upcycling” of
materials. This transformation concepts allows the “re-engineering” of the biomass to make
more valuable materials. Linking these two, we seek the conversion of waste resources into
high performing materials, fuels and ingredients. Some examples we can cite are:
• Production of bio polyesters from tomato and cork residues.
• Production of fuel intermediates from coffee waste and wood.
• Extraction of mucilaginous material from beans, seeds, and peels.
• Production of prebiotics from onions and artichokes.
• Production of polyphenols by lignin, black liquor or wood liquefaction.
• Production of antioxidants by ferulic-coumaric-lignin hydrolysis.
We can carry out such a diverse fan of applications thanks to the precise control over the
reaction time – temperature pair we apply to the reactor (severity). In this conference
presentation, we would like to show with examples the high flexibility of the technology by
correctly controlling time and temperature.

Keywords: Supercritical Water, Biorefinery, Fuels, Materials

29
 Simulation of heat transfer effects on the supercritical CO2
extraction at a two-vessel industrial plant
Felipe R. Toledo* and José M. del Valle

Departamento de Ingeniería Química y Bioprocesos, Pontificia Universidad Católica de Chile,

Santiago, Chile
*frtoledo@uc.cl

GRAPHICAL ABSTRACT

ABSTRACT
Simulation of a SuperCritical (SC) carbon dioxide (CO2) predicts the extraction process by
describing solute mass transfer from the solid substrate to the fluid. It can be used to predict
extraction yields in an industrial plant. However, heat transfer phenomena must be included in
calculations to account for temperature gradients formed within the extraction vessel in the
pressurization, extraction, and depressurization stages [1]. These temperature gradients affect
physical and transport properties. The depressurization and pressurization stages are relevant
in the industrial SC-CO2 extraction process because these plants use two or more extraction
vessels to approach a continuous operation by reconditioning one extraction vessel while the
remaining ones are in service [2]. The objective of this work was to simulate the heat transfer
in an SC-CO2 extraction process for a two-vessel industrial plant to assess the effect of those
temperature gradients on the rate and yield extraction.
We adapted the heat and mass transfer models and used the parameter values proposed by
Toledo & del Valle [1] to estimate temperature profiles at all stages of the industrial SC-CO2
extraction process. The modifications included temporal temperature variations at the
extraction stage and heat transfer within the vessel wall. The temperature of the fluid depends
on the extraction time and axial and radial position in the extraction stage, while in the
reconditioning stages (depressurization and pressurization), a perfect mixing is assumed and
only depends on the stage time (homogenous temperature). On the other hand, the temperatures
of the solid substrate and the vessel wall depend on the time and axial and radial positions in
all stages. We used as a case base a 370-L extraction vessel with a length-to-diameter ratio of
6, operating at 60 MPa, with a heating medium (service fluid) in contact with the external face
of the extraction vessel at 60 °C. In the vessel, 1-mm particles were extracted using 25 kg/min

30
of CO2 a 45 °C and 60 MPa (superficial CO2 velocity of 3 mm/s). We simulated cycles
(pressurization, extraction, and depressurization) starting from an extraction stage at isothermal
conditions (60 °C). At the end of the extraction stage, we averaged the final temperature of the
fluid (weighed average local temperatures by CO2 mass), and used this value as the initial
condition for the depressurization stage. On the other hand, we kept the local substrate and
vessel wall temperatures. In the pressurization stage, the initial vessel wall temperatures were
those at the end of the depressurization stage, and the substrate was at room temperature. We
simulated the heat transfer in these cycles until we reached a pseudo-steady state when
differences in local temperatures at the end of two consecutive extraction stages were
sufficiently small. After reaching this pseudo-steady state condition, we applied the mass
transfer model simultaneously with the heat transfer model, where we estimated local fluid
temperatures, which were used to estimate the fluid properties and mass transfer parameters.
Finally, we estimated the cumulative SC-CO2 extraction curves and compared them with those
obtained at isothermal extractions (using the average temperature among all the weighted
average temperatures along the extraction stage).
Results showed that the pseudo-steady state was reached in the third cycle of the process.
Temperature gradients affected cumulative SC-CO2 extraction curves at the pseudo-steady
state at industrial levels compared to those obtained in an isothermal extraction at 54 °C
(average temperature for all extraction time), obtaining a Mean Absolute Percentage Error of
10%. The impact of the fluid service and inlet CO2 temperatures on the average temperature of
the extraction vessel is related to the superficial CO2 velocity. At a higher CO2 velocity, the
average fluid temperature approaches the inlet CO2 temperature, while at lower CO2 velocities,
the average fluid temperature approaches the heating medium temperature.
In the following stages, process variables such as the CO2 and heating medium temperatures,
superficial CO2 velocities, and particle diameter will be modified to assess their effect on the
extraction rate and yield.

Keywords: Heat transfer, pressurization, depressurization, extraction, industrial plant.

REFERENCES
[1] F.R. Toledo, J.M. del Valle, Effect of heat transfer on the pressurization, extraction, and
depressurization stages of a supercritical CO2 extraction process. 1. Development and
validation of the heat transfer model, Submitted to J. Supercrit. Fluids. (2023).
[2] F.R. Toledo, J. García-Serna, G.A. Núñez, J.M. del Valle, Simulation of a flexible multi-
vessel extraction plant with counter-current contacting between a solid substrate and
supercritical CO2, Comput. Chem. Eng. 175 (2023) 108261.
https://doi.org/10.1016/j.compchemeng.2023.108261.

31
 Integrated and sustainable pressurized fluid processes for
biorefinery of plant and animal by-products
Marleny D.A. Saldaña*

Department of Agricultural, Food and Nutritional Science, University of Alberta

116 St & 85 Ave, Edmonton, AB, Canada.
*corresponding author email address: marleny.saldana@ualberta.ca

GRAPHICAL ABSTRACT

ABSTRACT
Every year, huge amounts of by-products from plants and animals are generated worldwide.
The improper disposal of these by-products results in global environmental concerns.
Therefore, proper management, development, and integration of technologies that are harmless
to the human being and protective of the environment have increased. Technologies that use
pressurized fluids such as subcritical water (sCW), pressurized ethanol and supercritical CO2
(SC-CO2) are considered “green” and environmentally friendly solvents. Research in my
laboratory has focused on the use of these pressurized fluids with various unit operations,
including extraction, fractionation, adsorption, reaction and particle formation to obtain
valueadded compounds from a variety of plant and animal by-products. Different plant by-
products, such as pomace, peel, husk, and straw, and animal by-products such as, shells and
feet, have been used to obtain valuable compounds.
In this presentation, some examples of high value-added compounds obtained after processing
with pressurized fluids will be demonstrated and discussed. Among the value-added
compounds obtained are sugars, phenolic acids, anthocyanins, amino acids, chitosan,
oligosaccharides, cellulose nanofibers among others. In one of the examples, a two-step
sequential approach was evaluated based on sCW technology and the use of citric and malic

32
acids to first extract and purify pea soluble polysaccharides from pea fiber, a co-product from
the food industry, and then convert them to pectic oligosaccharides. In another example, high
value-added compounds were obtained after sCW processing followed by SC-CO2 drying.
First, sCW technology was used to treat various agricultural by-products to obtain
hemicellulosic sugars and phenolic compounds, and a residue rich in cellulose and lignin. Then,
the obtained cellulose residue was converted to cellulose nanofibers. These nanofibers were
used to produce cellulose nanofiber hydrogels, and SC-CO2 drying was used to obtain cellulose
nanofiber aerogels. Additionally, examples using the biorefinery concept in the processing of
animal by-products aiming the production of new functional products and high value-added
co-products will be discussed. For example, we have treated shrimp shells using sCW
processing assisted by ultrasound to obtain value-added compounds such as chitin, chitosan
and amino acids that can be further use in several applications for the food, pharmaceutical and
biomedical sectors.
This presentation demonstrates that combining sCW, high-intensity ultrasound and SC-CO2
can increase process efficiency, and promote a sustainable economy. Overall, our results have
shown that pressurized fluids with various unit operations can be used for a variety of
applications.

Keywords: Subcritical water, Supercritical CO2, Unit operations, Reactions, Supercritical CO2
drying.

Acknowledgements
My research program at University of Alberta has been funded by the Natural Sciences and
Engineering Research Council of Canada (NSERC), Canada Foundation for Innovation (CFI),
Alberta Innovates, Alberta Pulse Growers and various Canadian and International companies.
I thank the 2023-2024 Killam Professorship Award, and the 2022-2023 Agricultural, Life and
Environmental Sciences Faculty International Engagement Award.

33
Processing of hemp seed and cannabis flowers using supercritical
fluids – a New Zealand perspective

Owen J. Catchpolea,*, Teresa Morenob, Campbell Ellisona, Stephen Bloorb

a
Callaghan Innovation (Food Processing Technologies), 1 Watt St, Auckland, New Zealand
b
Callaghan Innovation (Process Engineering), 69 Gracefield Road, Lower Hutt, New Zealand
*corresponding author email address: owen.catchpole@callaghaninnovation.govt.nz

GRAPHICAL ABSTRACT

ABSTRACT

Regulatory changes to New Zealand’s drug laws have now made it possible to establish a
medicinal cannabis product industry using locally grown cannabis, and hemp seed product
industry using hemp grown specifically to produce seed. Cannabis is grown under extremely
tightly regulated conditions, and the extracts produced must meet a minimum quality standard
before products can be sold by prescription. In contrast, hemp seed is less regulated, and any
food products are possible provided the seed is not viable. We have carried out R&D to
determine a suite of extraction, thermal processing and fractionation processes to convert hemp
and cannabis flowers through to partially purified cannabidiol (CBD). The feed material was
ground, dried and thermally decarboxylated for specified temperatures and times. The treated
flowers were then extracted with supercritical CO2 (with two stage pressure reduction) to give
a first extract enriched in CBD and wax-like material, and a second extract enriched in terpenes.
Comparative extractions have also been performed using ethanol, propane and dimethyl ether.
The first extract was then subjected to winterization using ethanol, filtration and vacuum

34
evaporation to remove solvent. Hemp seed processing in New Zealand has transitioned from
whole seed cold-pressing for hemp seed oil extraction to dehulling to give white coloured
hearts with high oil and protein contents and hulls as a by-product stream. A significant portion
of the hearts are green coloured (“chlorophyll hearts”), which are removed as a by-product by
colour sorting. In our work, the standard and chlorophyll hearts were ground and extracted with
supercritical CO2, subcritical propane or subcritical dimethyl ether (DME) to give high quality
oil and substantially fat free proteins. Comparative extractions were also performed on whole
seed hemp meal and hemp hulls. Further processing of the defatted seed hearts into hemp seed
protein hydrolysates and hemp milk were performed, along with extraction of phenolics
including cannabisins and residual oil from hemp hulls.

Keywords: Medicinal cannabis, hemp seed, supercritical extraction, cannabidiol, seed oil

35
 Ultra high-pressure processes-design and examples
Željko Kneza,b*, Volkmar Steinhagenc, Petra Kotnika, Maša Knez M.a
a
University of Maribor, Faculty of Chemistry and Chemical Engineering, SI-2000 Maribor, Slovenia,
b
CINS d.o.o, SI-2000 Maribor, Slovenia
c
Uhde High Pressure Technologies GmbH, D-58093 Hagen, Germany.
*corresponding author email address: zeljko.knez@um.si

GRAPHICAL ABSTRACT

ABSTRACT
Conventional solvents are potential environmental pollutants and the use of heat is
energyintensive, so research is focused on developing new processes with a lower
environmental impact. High-pressure technologies involving subcritical and supercritical
fluids offer the possibility of producing new products with specific properties or designing new
processes that are environmentally friendly and sustainable. The use of high pressure as a
process tool can also avoid regulatory restrictions on residual solvents and limitations on the
use of conventional solvents in chemical processes. The extraction of substances from plant
materials and their formulation "in situ" into products with specific properties is currently one
of the very promising applications of supercritical fluids.
There are several processes using sub- and supercritical fluids that have already been developed
for commercial scale, dry cleaning, high-pressure sterilization, jet cutting, thin film deposition
for microelectronics, separation of value-added products from fermentation broths in the field
of biotechnology, and as a solvent in syntheses. All these applications lead to sustainable
production methods that are not only more environmentally friendly but also give products
very special properties.

36
The design of all high-pressure processes requires information on operating parameters, solvent
type and quantity, recirculation rate and energy consumption. This information can be obtained
by phase equilibrium and mass transfer measurements. Therefore, before selecting the
appropriate treatment method, several parameters must be taken into account which affect the
solubility, mass transfer of the target compounds in the sub- or supercritical fluid and
consequently the extraction yield. The quality of the extract depends on pressure and
temperature, which can have a serious influence on the composition of the final extracts. In
addition, the effect of pressure drop in extraction material should be evaluated and taken into
account in the optimization of the parameters in order to achieve the best ratio between yield
and solvent volume, and extraction time. There is an additional requirement, namely the
maximum possible subcritical or supercritical solvent loading to be achieved in the extraction
phase of the processes, while in the separation phase, the solute solubility in the solvent should
be the lowest.
Since a high solubility of the desired compound in the supercritical solvent is essential for the
economy of the extraction process, it should be verified by practical analyses whether
extraction using supercritical fluid is a suitable technique for the isolation of the target
compound or whether conventional solvents should be used.
Particular attention should be paid to the thermodynamic basis of these processes – phase
equilibrium data for systems of plant extracts or pure substances with various gases such as
carbon dioxide and more recently propane, argon, chlorinated hydrocarbons, and sulfur
hexafluoride.
In addition to the assessment of basic thermodynamic data, transport data describing the
behavior of the system under certain conditions, such as density, viscosity, dielectric constant,
diffusion coefficient, and interfacial tension, must also be considered. A range of these data,
measured at different pressures and temperatures, is available in the scientific literature for a
number of pure compounds. Data on the behavior of binary systems at very high pressures and
temperatures are still relatively scarce and involve methods that are either expensive or
timeconsuming.
A comparison of the composition and yield of the products obtained from low and ultra-
highpressure extraction will be presented. One of the most important advantages of using
subcritical or supercritical fluids at ultra-high pressure is the highly selective extraction of
components or fractionation of whole extracts. The unique properties of SCF, especially at
ultra-high pressure, can be used for fractionation of extracts, particle formation, impregnation
of solid particles, formation of solid powder emulsions, coating of particles, e.g. for the
formation of solids with unique properties to be used in a variety of applications.
Results of our research on thermodynamics, mass transfer and extraction processes carried out
in the pressure range up to 1500 bar and above will be presented.

Keywords: Ultra high pressure, design data, industrial plant, herbs extracts

37
 Development of an Online Integrated Method for Analyzing
Bioactive Compounds: Pressurized Liquid Extraction Coupled
with High-Performance Liquid Chromatography
Juliane Viganóa,b,*, Monique Martins Striederb, Rodrigo S. Pizanib, Letícia S. Contierib,
Leonardo M. de Souza Mesquitab, Mauricio A. Rostagnob
a
Centro de Ciências da Natureza, Universidade Federal de São Carlos, Rod. Lauri Simões de Barros,
km 12 – SP 189, Buri, SP 18290-000, Brazil
b
Multidisciplinary Laboratory of Food and Health (LabMAS), School of Applied Sciences (FCA),
Universidade Estadual de Campinas, São Paulo, Brazil
*julianevigano@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
The accurate analysis of biocompounds, such as polyphenols in solid samples, relies heavily
on effective solid-liquid extraction methods. Various extraction techniques have been
employed to prepare samples prior to chemical analysis. Furthermore, the specific analytical
method employed varies depending on the target analyte. Historically, sample preparation and
chemical analysis have been conducted using offline methods, meaning that extraction and
chemical analysis were carried out separately, without integration. As a result, the development
of a single commercially available equipment integrating extraction and analysis has remained
unfeasible due to numerous inherent challenges. Overcoming these obstacles and fostering the
advancement of more sustainable analytical procedures require the adoption of innovative
techniques.
Pressurized liquid extraction (PLE) is a well-established technique for sample preparation in
the analytical step and has been extensively studied for a wide range of raw materials. One of
its key advantages is the utilization of pressurized solvents, enabling the liquid phase to exceed
its normal boiling point. This significantly enhances mass transfer, extraction rates, and reduces
overall extraction time. In terms of chemical analysis, high-performance liquid
chromatography (HPLC) stands out as one of the primary techniques employed.

38
Therefore, the objectives of this study are to describe a novel online integrative system,
combining pressurized liquid extraction (PLE) with high-performance liquid chromatography
(HPLC) using a 2D PLE·HPLC-PDA system, and to present its validation for two different
solid samples. The introduction of this innovative online method marks a significant
advancement in the field of extraction and analysis automation systems, taking us a step closer
to the future of streamlined and efficient analytical processes.
The system consists of two modules: one for pressurized liquid extraction (PLE) and the other
for high-performance liquid chromatography (HPLC). These modules are integrated using an
interface valve, which is programmed to separate a portion of the extract (20 µL) for injection
into the HPLC module. During each run, solid samples weighing 0.5 g were loaded into the
PLE module, which was set to a temperature of 70 ± 1 ºC. The extraction process commenced
by flowing a solvent (70 vol.% ethanol) at a rate of 2 mL/min, under a pressure of 1500 ± 50
psi. At regular intervals (every 5 minutes), automatic injections of the extract were performed
into the HPLC module. The remaining extract that was not injected into the HPLC was
collected in fractions at the end of the PLE module. The chromatographic methods were
adjusted according to the type of solid sample being analyzed. To validate the system, readily
available coffee and passionfruit byproducts were utilized as sources of bioactive compounds.
Passionfruit bagasse and coffee husks were employed as solid samples, with each PLE process
taking approximately 120 minutes per sample.
The collected extract fractions were subjected to offline analysis that consisted of the manual
injection the extract sample into the HPLC. The online and offline analysis evaluated the
extraction yield of piceatannol, chlorogenic acid, and caffeine for passion fruit bagasse and
coffee husks, respectively.
A comparison was conducted between the extraction yields obtained from online and offline
injections. The results showed that the extraction yields were not significantly different (p >
0.05). However, it is important to acknowledge certain limitations of the system. For each
individual sample, the timing for the first online injection should be carefully evaluated. The
mass of the sample and the flow rate of the extraction solvent must be adjusted considering the
target compound content and the calibration curve range. Opting for a shorter chromatographic
method can minimize discrepancies between online and offline quantifications, making shorter
methods preferable. Additionally, the compatibility of the extraction solvent and mobile phase
should be considered.
Overall, the 2D PLE·HPLC-PDA system demonstrates its feasibility as a tool for integrating
solid sample preparation and chemical analysis of biocompounds into a single online step.

Keywords: high-pressure technologies, extraction, sample characterization, green chemistry,

bioactive compounds.

39
 COME INTO A NEW ERA OF PURIFICATION

Assia Konea, Piotr Kesickia

a
SFE Process
*
E-mail: akone@sfe-process.com

GRAPHICAL ABSTRACT

ABSTRACT
Step into a new era of supercritical purification with SFE PROCESS, a leading manufacturer
in cutting-edge extraction technology. In this presentation, we explore the transformative
potential of supercritical fluid extraction (SFE) and chromatography (SFC). At SFE PROCESS
we are pioneers in sustainable and precise extraction and purification methods that elevate
industries like pharmaceuticals, cosmetics, and the food industry. Discover the benefits of
supercritical technologies, including enhanced selectivity, minimal residue, environmental
sustainability. With a focus on efficiency and improved product quality, these innovations
redefine industrial processes. Join us to unlock the future of extraction and purification and
embrace a cleaner, greener, and more efficient approach with SFE PROCESS.

Keywords: Supercritical, Purification, Extraction

References
[1] Chen, Y.-W., Hsieh, D.-J., Periasamy, S., Yen, K.-C., Wang, H.-C. and Chien, H.-H. (2021),
Development of a decellularized porcine bone graft by supercritical carbon dioxide extraction
technology for bone regeneration. J Tissue Eng Regen Med, 15: 401-414.
https://doi.org/10.1002/term.3181
[2] J. Fages, A. Marty, C. Delga, J-S Condoret, D. Combes, P. Frayssinet. (1994), Use of
supercritical CO2 for bone delipidation. Biomaterials 15: 650-656. https://doi.org/10.1016/0142-
9612(94)90162-7

40
 Complex Fluid Phase Behavior in Binary and Ternary Systems,
including the Critical Region

Cor J, Petersa , Kerstin Gauterb

a
Colorado School of Mines, Department of Chemical and Biological Engineering,
Center For Hydrate Research, Colorado, Golden CO 80401, USA
b
Delft University of Technology, Faculty of Applied Sciences,
Laboratory of Applied Thermodynamics and Phase Equilibria,
Julianalaan 136, 2628 BL Delft, The Netherlands
*corresponding author email address: cjpeters2006@gmail.com

ABSTRACT
For practical applications of near-critical fluids, e.g., separations, reactions, materials
processing, etc., in all cases a thorough knowledge of the phase behavior of such systems is
a prerequisite. Although carbon dioxide is an abundant, inexpensive, nontoxic, and
nonflammable solvent, in supercritical fluid applications it has the major disadvantage of
having a poor solvency with respect to many low-volatile components, and cosolvents like,
for instance, alkanols must be used to enhance the solubility of the solutes in supercritical
carbon dioxide.
Another characteristic feature of carbon dioxide in supercritical fluid applications is that in
many cases the fluid mixture is composed of, on the one hand, a highly volatile near-critical
solvent (carbon dioxide) and, on the other hand, one or more low-volatile solutes, i.e., in
practice we are dealing with mixtures composed of physically and/or chemically very
dissimilar molecules. From many investigations, it is known that in these kinds of mixtures
the presence of a second liquid phase is most likely to occur. The latter phenomenon also
makes carbon dioxide a less attractive near-critical solvent.
Besides the occurrence of a second liquid phase, from extensive and systematic experimental
research it became apparent that in binary mixtures of near-critical carbon dioxide with
certain low-volatile solutes, belonging to a homologous series of substances, the nature of
this multiphase behavior may undergo major changes. For instance, the nature of the fluid
multiphase behavior in the binary mixture of near-critical carbon dioxide with dodecane is
completely different compared to that with tetradecane. Although in both cases a three-phase
equilibrium liquid-liquid-vapor (l1l2g) is present, their critical endpoints differ in nature. In
the carbon dioxide system with dodecane the critical endpoint of the three-phase equilibrium
l1l2g is characterized by criticality of the two liquid phases l1 and l2 (l1 = l2 + g), while in the
critical endpoint of the three-phase equilibrium l1l2g of the carbon dioxide system with
tetradecane the liquid-phase l1 is critical with the gaseous phase. The fluid multiphase
behavior of the system near-critical carbon dioxide with tridecane is characterized by the
presence of even three critical endpoints: two of them being of the nature l 1= l2 + g and the
other one showing the phenomenon l1= g + l2. In all, this demonstrates that minor changes in
the size and/or shape of the solute molecules sometimes may cause major changes in its phase
behavior. A closer look at the various phase diagrams of these binary mixtures will show that
in each of the three systems mentioned above the connectivity of the critical loci is different,
i.e., each system belongs to a different class of fluid phase behavior. From detailed
experimental studies on the fluid multiphase behavior of these so-called asymmetric systems,

41
 it became apparent that the phenomena described above are not an exception but apply in
general.
Another phenomenon of interest for supercritical fluid technology is the occurrence of co-
solvency effects in ternary mixtures of a near-critical solvent and two different solutes. For
instance, addition of a small amount of a 1-alkanol to a binary mixture carbon dioxide + n-
alkane may cause a significant effect on the critical behavior of the system with consequently
unexpected phase behavior like miscibility windows and two-phase holes in three-phase
surfaces. In general, miscibility windows occur at higher pressures than holes. Although it is
expected that both phenomena may have a consider-
In this contribution, we review the occurrence of so-called holes as one of the complexities
in fluid multiphase behavior that may occur in ternary mixtures of near-critical carbon
dioxide and two low-volatile solutes e.g., in certain combinations of n-alkanes, 1-alkanols,
and o-nitrophenol. It is believed that the selected systems are representative for systems to
be met in supercritical fluid applications. From this study it became apparent that in the
region of interest small fluctuations in the concentration of the components the type of fluid
phase behavior in the sense of the Scott and van Konynenburg classification may change
several times, or in other words, the number of coexisting phases may change several times
from two into three and vice versa. For obvious reasons this makes the design of processes
with near-critical carbon dioxide extra complicated since it may be expected that the
phenomena discussed in this review are very general and not only limited to the solutes
investigated in this study.
From this study it also could be concluded that in the transformation schemes with a type-I
or type-V fluid phase behavior system involved an inconsistency in the ternary phase
diagram may arise. It has been established that the inconsistency caused by the presence of
a type-I sub-binary system in the ternary system only can be removed by considering the
type-I system being a type-II system. A similar inconsistency occurs if the ternary system
has a type-V sub-system. In this case the sub-binary system only can transform to a type-IV
sub-system.

Keywords: Phase Equilibria, Critical Phenomena, Ternary Systems

42
 ScCO2-impregnation of clove extract in polycarbonate, poly(L-
lactic acid) and their blends

Amanda M. Jordãoa, Jorge A. P. Monteiroa, B. Grignardb, C. Jérômeb, Eric K. Silvac, Lígia. P.

Maia-Obia, Bruno. G. da Silvad, Mathilde Champeaua,*
a
Center of Engineering, Modelling, and Applied Social Sciences, Federal University of ABC, Santo
André, Brazil
b
Center of Education and Research on Macromolecules, Université de Liège, Belgium
c
School of Food Engineering, University of Campinas, Campinas, Brazil
d
Center for Natural and Human Sciences, Federal University of ABC, Santo André, Brazil
*Corresponding author: mathilde.champeau@ufabc.edu.br

GRAPHICAL ABSTRACT

ABSTRACT
Polymers able to release natural extracts are relevant systems in many fields such as food
packaging, wound care, cosmetics, and agriculture. The polymer protects the extract from
degradation, and exhibits the extract bioactivity. The supercritical CO2-assited impregnation
(SCI) process is a promising alternative to other manufacturing processes to prepare extract-
releasing polymeric products, since it operates at mild temperatures and allows recovering
products free of solvent. While the SCI of a single molecule in a polymer is well understood,
working with multicompound extracts (natural extracts) is challenging due to their complex
composition. The bioactivity of a natural extract may be attributed to a specific compound or
group of compounds. Consequently, the challenge of scCO2 impregnation of natural extracts
is not limited to controlling the total loading. It is also essential to ensure that the loaded

43
extract's composition may preserve the crude extract's bioactivity, which can be ensured by
impregnating the compound(s) responsible for the desired bioactivity. Thus, it is crucial to
evaluate the influence of the operational conditions on the composition of the impregnated
extract, in addition to the total loading.
The goal of this research project was to investigate the specific phenomena involved in the
impregnation of a mixture of compounds, and the consequence on the modification of natural
extracts’ composition once impregnated. Herein, we focused on the impregnation of clove
extract (CE) into polycarbonate (PC) and poly(L-lactic acid) (PLLA), as well as in their blends.
CE (Eugenia caryophyllus) was obtained via supercritical fluid extraction (SFE) from flower
buds, being rich in eugenol, and containing eugenyl acetate, β-caryophyllene, and α-humulene.
CE is known for its antibacterial, antifungal, and insecticidal properties, similar to synthetic
preservatives. PC and PLLA were chosen since their CO2 absorption is known, they possess
different thermal transitions and their distinct chemical structures imply different affinities for
the four main compounds of CE. First of all, a face-centered design was applied to evaluate the
influence of the temperature (35, 47.5, and 60 ºC) and pressure (10, 20, and 30 MPa) on the
clove loading (CL%) and on the composition of the loaded extract in PC and PLLA. The
amount of impregnated extract and its composition in the polymeric matrix were determined
gravimetrically and by gas chromatography, respectively. For both PC and PLLA, significant
CL% were achieved (6.8 – 44%). In the case of PC, the temperature favored the impregnation
whereas the pressure was deleterious, whereas no clear tendency was observed for PLLA and
it melted in some conditions due to saturation with CE. The composition of the extract
impregnated in PC and PLLA was different from the crude extract, suggesting the selectivity
of the impregnation process. However, similar composition of the impregnated extract were
noticed in both polymers. The eugenol content increased from 70.06 to the range of 81.31-
86.28 %. The eugenyl acetate, β-caryophyllene, and α-humulene contents decreased from 17.06
to 11.22 – 14.99 %, 10.67 to 1.03 – 4.83 %, and 2.20 % to below 0.51, respectively. In PC, the
pressure enhanced the eugenol content whereas it decreased the eugenyl acetate content in the
loaded extract, whereas the temperature had the opposite effect. The results were rationalized
using the Hansen type solubility parameters and the intermolecular interactions between the
polymers and each compound. Differential scanning calorimetry (DSC) showed a high
plasticizing effect of CE on both polymers, which is responsible for an increased chain mobility
that is favorable to the impregnation. In the second part of the project, immiscible blends of PC
and PLLA were produced with different PC/PLLA ratio. The samples were characterized by
DSC, scanning electron microscopy, and dynamic mechanical analysis. The influence of the
blend composition and of their microstructure on CL% was rationalized.
This study has highlighted the selectivity of the scCO2-impregnation process on
multicompound extracts. Using blends and controlling the microstructure can be a relevant
strategy to control the impregnation yield and the functional properties of the impregnated
polymer. The impregnated polymers are promising for antibacterial food containers application
and active implants due to the high loading of clove extract and the high eugenol content.

Keywords: Supercritical Solvent Impregnation, Blend, Phase separation, Natural Extract

44
 Selection of green extraction solvents using the Hansen solubility
parameters approach
Andrea del Pilar Sánchez-Camargo

Grupo de Diseño de Productos y Procesos (GDPP), Department of Food and Chemical Engineering,
Universidad de los Andes, Carrera 1E No. 19 A 40, Bogotá, Colombia.
Email address: ad.sanchez@uniandes.edu.co

GRAPHICAL ABSTRACT

ABSTRACT

In today's context, green extraction processes are primarily oriented towards the development
of innovative strategies to attain sustainable separation methods. These methods advocate the
utilization of alternative solvents derived from renewable sources. Green solvents have
emerged as a viable option to replace petroleum-based counterparts. However, it is imperative
that they maintain equivalent extraction properties and technological attributes. The selection
of an appropriate solvent stands as a pivotal phase in the process of substituting solvents.
Hence, the Hansen solubility parameters (HSP) have proven to be an effective decision-making
tool. This approach boasts several noteworthy advantages, including its simplicity and efficient
utilization of time and resources.
During this plenary conference, we will elucidate the factors influencing the solvent selection
process from both a chemical and structural standpoint. This discussion encompasses scenarios
involving both subcritical and supercritical states. Furthermore, we will provide an overview
of the application of HSP as a solvent selection tool for extracting bioactive compounds from
natural products. Additionally, we will present a case study that predicts the extraction of a
carotenoid (fucoxanthin) from microalgae using green solvents, employing the HSPIP®
software [1-4].

45
Keywords: Alternative solvents, Bioactive compounds, Green extraction, Hansen solubility
parameters

References :

[1] Sánchez-Camargo, A.P., Bueno, M., Ballesteros-Vivas, D., Parada-Alfonso, F., Cifuentes,
A., Ibañez, E. 2.46 - Hansen Solubility Parameters for Selection of Green Extraction Solvents,
Editor(s): Alejandro Cifuentes, Comprehensive Foodomics, Elsevier, 2021, Pages 710-724,
ISBN 9780128163962.
[2] dos Santos, L.C.; Mendiola, J.A.; Sánchez-Camargo, A.d.P.; Álvarez-Rivera, G.; Viganó,
J.; Cifuentes, A.; Ibáñez, E.; Martínez, J. Selective Extraction of Piceatannol from Passiflora
edulis by-Products: Application of HSPs Strategy and Inhibition of Neurodegenerative
Enzymes. Int. J. Mol. Sci. 2021, 22, 6248. https://doi.org/10.3390/ijms22126248
[3] Pagano, I., Sánchez-Camargo, A., Mendiola, A.J., Campone, L., Cifuentes, A., Rastrelli,
L., Ibañez, E. Selective extraction of high-value phenolic compounds from distillation
wastewater of basil (Ocimum basilicum L.) by pressurized liquid
extraction. Electrophoresis 2018, 39, 1884–1891
[4] Sánchez-Camargo, A.P, Natalia Pleite, Herrero, M., Cifuentes, A., Ibáñez, E., Gilbert-
López, B. New approaches for the selective extraction of bioactive compounds employing bio-
based solvents and pressurized green processes, The Journal of Supercritical Fluids, 2017, 128,
112-120. https://doi.org/10.1016/j.supflu.2017.05.016.

46
 ORAL
COMMUNICATIONS

47
 Simultaneous extraction of macauba oil and active compounds
from olive leaves using pressurized propane
Jonas M. Jaskia, Carlos E. Barãoa,b, Camila da Silvad, Lúcio Cardozo-Filhoa,d, Vladimir F.
Cabralc
a
Postgraduate Program of Agronomy, State University of Maringá, Av. Colombo 5790, Maringá,
Brazil
b
Federal Institute of Paraná, Campus Paranavaí, Rua José Felipe Tequinha, 1400, Paranavaí, Brazil
c
Department of Food Engineering, State University of Maringá, Av. Colombo 5790, Maringá, Brazil
d
Department of Chemical Engineering, State University of Maringá, Av. Colombo 5790, Maringá,
Brazil
*corresponding author email address: vfcabral@uem.br

GRAPHICAL ABSTRACT

ABSTRACT
Olive leaves are olive-growing products with high commercial value because they have
valuable bioactive components. Macauba seeds have different characteristics with potential use
in the food agroindustry. The simultaneous extraction of plant matrices (seeds and leaves) can
incorporate substances into the oils in the same process. The pressurized n-propane solvent can
be used to obtain solvent-free oils from plant matrices. Moreover, recent works showed that
these oils could act as co-solvents in extractions of bioactive compounds. In this context, the
main goal of this work was to obtain high-quality oils with attractive nutritional properties and
high levels of active compounds by obtaining extracts from macauba seeds and olive leaves in
an only structured bed using propane as a solvent. In the first section of the structured bed,
propane extracts the oil from the macauba seeds, and in the next section, the mixture of propane
+ oil (co-solvent) extracts bioactive components from olive leaves. The percentage mass yield,
fatty acids (FA), bioactive compounds, and oxidative stability were analyzed. The percentage

48
mass yield of the oils obtained using or not using olive leaves was 26.0% and 32.3%,
respectively. Macauba oil obtained here not using olive leaves had the highest percentage of
saturated FA, mainly lauric acid (28%); among the unsaturated FA, oleic acid (32%) was the
major. The oils produced using the olive leaves did not have their profile of fatty acids altered,
but there was the aggregation of bioactive compounds from the olive leaves to the oil, mainly
α-tocopherol (0 – 92 mg•100g-1), campesterol (4.6 – 5.2 mg•100g-1) and β – sitosterol (35.6 –
131.7 mg•100g-1). Oxidation induction time increased from 9.2 to 13.2 hours for the oil
obtained using olive leaves. Such results show that incorporating bioactive compounds from
olive leaf extract in a vegetable oil of macaúba, using propane as a solvent, promotes increased
resistance to lipid oxidation and more active compounds in oils.

Keywords: bioactive compounds, phytosterols, pressurized fluid, lipid oxidation.

49
 Development of β-sitosterol-loaded nanocapsules obtained by
supercritical fluid extraction of emulsion process for potential use
in functional foods
Daniela Cerroa,b, Camilo Streitta,b, Adrián Rojasa,c,Julio Romerob, Alejandra Torresa,c,*
a
Center for Packaging Innovation (LABEN), Food Science and Technology Department,
Technological Faculty, University of Santiago de Chile, Chile.
b
Laboratory of Membrane Separation Processes (LabProSeM), Chemical Engineering Department,
Engineering Faculty, University of Santiago de Chile, Chile.
c
CEDENNA, Center for the Development of Nanoscience and Nanotechnology, Chile
*corresponding author email address: alejandra.torresm@usach.cl

GRAPHICAL ABSTRACT

ABSTRACT
β-sitosterol has demonstrated hypocholesterolemic effects for the prevention of cardiovascular
diseases. However, its application as a bioactive compound is limited due to its low water
solubility and poor bioavailability. One of the strategies to overcome these limitations involves
its encapsulation based on biopolymers. The supercritical fluid extraction of emulsions (SFEE)
process is one of the most widely used encapsulation techniques in recent years. This process
uses supercritical CO2 to remove the organic solvent from an emulsion at moderate process
temperatures, protecting the compound to be encapsulated. In turn, SFEE process allows the
development of micro o nanoscale sizes that further improve the final product properties. The
objective of this work was to study the effect of process variables (CO2 pressure and operation
time) on the physical-chemical properties of nanocapsules loaded with b-sitosterol obtained by
supercritical fluid extraction of emulsion (SFEE) process using polycaprolactone and modified

50
starch as coating material for their potential use in functional foods. For this purpose,
nanoemulsions of acetone in water (o/w) was prepared in a 20/80 v/v ratio for a total volume
of 50 mL. The nanoemulsions were obtained by homogenization with ultrasound assistance.
0.25 %w/w of surfactant tween 80 content was used, 0.5 %w/w of modified starch content or
0.4 %w/w of polycaprolactone content were used and ultrasonic time between 5-20 min.
Droplet size were measured in dynamic light scattering equipment, in addition to physical
stability for 90 min. Subsequently, the emulsions were put through a step of acetone solvent
elimination using supercritical CO2 to 9 and 13 MPa and time from 30 to 180 min at 40°C for
the capsules precipitation and subsequent freeze-drying, obtaining powders that were
characterized in terms of encapsulation efficiency, in addition for their physicochemical,
structural and thermal properties. The formulated emulsions serving as the starting point for
the SFEE encapsulation step were about 390 nm droplet size and were stable to storage. At 9
MPa and operation time of 90 min, higher encapsulation efficiency around 70-80% and more
defined core-shell morphologies and smaller particle size were obtained. In addition, the
structural and thermal properties confirmed the powders stability with the incorporation of β-
sitosterol. In conclusion, it were successfully developed β-sitosterol-loaded nanocapsules
obtained by supercritical fluid emulsion extraction process using polycaprolactone and
modified starch as coating material and have a greater potential for the development of a
product with potential use in functional foods or nutraceutical products.
Keywords: Nanoencapsulation, β-sitosterol, hypocholesterolemic effects, Supercritical Fluids,
Process Variables.

51
 Concentration of oxygenated terpenes in orange essential orange
by supercritical fluid adsorption
Victor Saldanha Carvalhoa*, Juliane Viganóab, Luis Henrique Fasolina, Julian Martineza
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas, -SP,
Brazil.
b
Centro de Ciências da Natureza, Universidade Federal de São Carlos, Rod. Lauri Simões de Barros,
km 12 – SP 189, Buri, SP 18290-000, Brazil
*Corresponding author email address: victorsalcarvalho@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT

The orange essential oil is used in several areas, such as the food, chemical and pharmaceutical
industries, working as a flavoring agent and fragrance ingredient in cosmetics and herbal
medicines. The major oxygenated terpenes found in this oil are linalool and terpineol, which
are responsible for their functional properties and aroma. To increase these functional
properties and the lifetime of the orange oil, many technologies to increase the oxygenated
monoterpenes have been studied, like supercritical fluid adsorption (SFA). SFA consists of
putting a supercritical mixturein contact with an adsorbent under controlled temperature and
pressure conditions. In this process, the compounds of the mixture are selectively adsorbed.
Once the adsorption is complete, the adsorbed compounds are gradually released in the final
fractions, significantly enhancing their concentration. Therefore, the objective of this work was
to concentrate linalool and terpineol in an orange oil fraction by supercritical fluid adsorption
(SFA) using silica aerogel as an adsorbent. The orange oil was donated by the company
Citrosuco®, located in Matão-SP, Brazil and the silica aerogel was bought from BuyAerogel®,

52
Cabot Enova® IC3100 Aerogel. SFA consisted in two steps: i) Continuous fractionation, where
sc-CO2 preferentially solubilizes short-chain nonpolar monoterpenes from orange oil; ii)
Adsorption, where the mass transfer of linalool and terpineol from the supercritical solution to
the adsorbent takes place. In the second step, the sc-CO2 stream with the solubilized orange oil
interacts with the adsorbent, promoting the contact and interaction of the orange oil
monoterpenes with the silica aerogel. In both steps, a 3² complete factorial design was
performed, varying temperature (40, 50, and 60 ºC) and pressure (10, 17, and 24 MPa). The
lower and upper limits of temperature and pressure were chosen considering the capacity of
the equipment and the thermal sensibility of monoterpenes at temperatures above 60 ºC.
Around 10 g of oil were used in both steps. The sc-CO2 flow rate was 4.2 g/min. For the second
step, column filled with 1.5 g of silica aerogel was coupled to the equipment immediately after
to the solubilization cell. Orange oil fractions were collected every five minutes along the
process. Ethyl acetate was used to adsorb the aerogel monoterpenes, and the elutions were sent
for gas chromatography analysis. The responses evaluated in SFA were the oil recovery, the
concentration and purification factors of linalool and terpineol, and the concentration of these
compounds adsorbed on the aerogel. Continuous fractionation achieved an oil recovery of
approximately 70% under conditions of 24 MPa/60 ºC, 24 MPa/50 ºC, 24 MPa/40 ºC and 17
MPa/60 ºC. However, under the performed conditions, the sc-CO2 solvating power was not
selective for the oxygenated monoterpenes concentration. However, continuous fractionation,
without the use of the adsorbent, obtained average concentration factors of 2.5 for linalool, 2.3
for terpineol and 1.9 for limonene. This indicates a promising strategy for the previous
concentration step, as the selectivity of sc-CO2 itself was able to concentrate the monoterpenes
by not solubilizing the non-volatile compounds present in cold-pressed orange essential oil.
Myrcene was the only monoterpene that had its concentration reduced, possibly because it is
the most volatile and has been carried out with CO2 in depressurization. In the second step,
when added an adsorbent column, the process was selective for oxygenated monoterpenes
concentration. Achieving concentration of 3.10 and 7.54, and purification factors of 2.2 and
4.9, for linalool and terpineol, respectively. Differently from the continuous fractionation, the
adsorption process involves intermolecular interaction, which makes the process more
selective. These results were obtained at the condition of lowest pressure and highest
temperature, in the fraction collected between 80 and 85 min at 10 MPa and 60 ºC. However,
this fraction represented only 6% (g/g) of oil recovery. The sc-CO2 is used as a solvent to
promote mass transfer from the oil to the aerogel, when the temperature increases, the mass
transfer also increases, resulting in the highest concentration and purification factor for the
higher temperature (60 ºC). We also observed that at 10 MPa and 60 ºC the lowest dynamic
solubility (15,8 ± 1,1 g oil/kg CO2) was obtained. This lower solubility resulted in a longer
process time (85 min) in relation to the other conditions and constant and lower oil recovery
rate. Contrary to those conditions that resulted in higher solubilities, in which in 40 minutes of
the process all oil had already been recovered. Corroborating the results, the concentrations of
terpenes adsorbed in the condition 10 MPa and 60 ºC were the highest. Therefore, it can be
concluded that SFA with sc-CO2 at low solvation power conditions is indicated for
concentrating oxygenated monoterpenes in orange oil.
Keywords: Supercritical fluid adsorption, orange essential oil, concentration, oxygen terpenes,
bioactive compounds.

53
Supercritical CO2 and melt-extrusion to produce enhanced PLGA
scaffolds
I. Gracia a, F. Carrascosa a*, J.M. García a, M.J. Ramos a, J.F. Rodríguez a, M.T. García a.
a
University of Castilla-La Mancha, Avda. Camilo Jose Cela, 12, 13071, Ciudad Real, Spain.
*corresponding author email address: Fernando.Carrascosa@uclm.es.

GRAPHICAL ABSTRACT

ABSTRACT
Traditionally, bone implants have been made with metals or their alloys, but they have some
problems and complications. However, these issues might be resolved by porous scaffolds
made of biocompatible and biodegradable polymers. There are various polymers proposed for
this application, among them poly(lactic-co-glycolic acid) (PLGA) stands out as highly
biodegradable and bioabsorbable polymer, in fact, it has been approved by the Food and Drug
Administration [1].
However, this polymer's weak mechanical strength is one of its main problems [2]. Many
studies have set their goals to reinforce these porous scaffolds by introducing reinforcing agents
into the polymer matrix. Various compounds have been used as reinforcing agents as different
calcium phosphates, Mg and derivates [3–5]. Additionally, according to some papers, using
these compounds enhanced the polymer to issues like acidic degradation or poor
osteointegration, suggesting that they may work in concert to improve scaffolds.
PLGA 75/25 was mixed with Mg, Mg(OH)2 and hydroxyapatite through melt-extrusion, later,
each composite and pure PLGA were foamed with supercritical CO2 under conditions ranging
from 100 to 200 bar of pressure and 25 to 60 ºC of temperature. Mechanical strength of
manufactured foams was measured through compression test and compared parameters as

54
Maximum strength. Other important parameters were also measured as main pore size by SEM
images, or porosity by 3D scanner.
Foam development with homogeneous dispersion of the added agents was demonstrated. In
addition, the mechanical properties of foams containing reinforcing agents were improved
compared to pure PLGA foams.

References

[1] X.H. Zhu, L.Y. Lee, J.S.H. Jackson, Y.W. Tong, C.H. Wang, Biotechnol. Bioeng. 100
(2008) 998–1009. https://doi.org/10.1002/bit.21824.
[2] E. Salonius, V. Muhonen, K. Lehto, E. Järvinen, T. Pyhältö, M. Hannula, A.S. Aula, P.
Uppstu, A.M. Haaparanta, A. Rosling, M. Kellomäki, I. Kiviranta, J. Tissue Eng. Regen.
Med. 13 (2019) 406–415. https://doi.org/10.1002/term.2801.
[3] M.O. Montjovent, L. Mathieu, H. Schmoekel, S. Mark, P.E. Bourban, P.Y. Zambelli,
L.A. Laurent-Applegate, D.P. Pioletti, J. Biomed. Mater. Res. - Part A. 83 (2007) 41–
51. https://doi.org/10.1002/jbm.a.31208.
[4] A. Brown, S. Zaky, H. Ray, C. Sfeir, Acta Biomater. 11 (2015) 543–553.
https://doi.org/10.1016/j.actbio.2014.09.008.
[5] T.M. Bedair, C.K. Lee, D.S. Kim, S.W. Baek, H.M. Bedair, H.P. Joshi, U.Y. Choi, K.H.
Park, W. Park, I.B. Han, D.K. Han, J. Tissue Eng. 11 (2020).
https://doi.org/10.1177/2041731420967591.

55
 High-pressure sequential extractions using aqueous solutions of deep
eutectic solvents (DES): a sustainable process with Green Certificate
Laís Benvenuttia*, Acácio Antonio Ferreira Zielinskia, Sandra Regina Salvador Ferreiraa
a
Chemical and Food Engineering Department, Federal University of Santa Catarina, C.P. 476, 88040-
900, Florianopolis, Brazil
*corresponding author email address: labenvenutti@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT

The growing world population demonstrates the need to develop economically viable processes
to provide the food demand. Besides, over the last decade, there has been a growing interest
from global consumers for natural compounds, as ingredients and additives to different
products, in replacement to synthetic ones, attending health and sustainability concerns. In this
sense, the plant-based extraction processes, mainly focusing on the valorization of industrial
by-products, can be an economic, ecological, and sustainable solution. Green processes reduce
energy consumption and the use of fossil-based solvents, ensuring the security and quality of
the recovered extract. Therefore, using aqueous solutions of deep eutectic solvents (DES) as
novel green solvents, coupled with high-pressure methods, emerging as a potential eco-friendly
alternative to fractionate different target compounds from vegetal matrixes. This novel
approach was applied for the valorization of jaboticaba peel. Pressurized DES aqueous
solutions were used in sequential extractions to recover the anthocyanin- and the pectin-rich
fractions from jaboticaba peel, following previously optimized conditions. Initially,
Pressurized Liquid Extraction (PLE) was applied to obtain the anthocyanin-rich fraction using
a 47% DES solution of choline chloride and malic acid (ChCl:Ma) at 10 MPa, 90 °C, and a

56
flow rate of 5.3 mL/min for 12 min. Then, Supercritical Water Extraction (SWE), using water
modified by 8% DES composed of citric acid, glucose, and water (Ca:Glu:Wa), was employed
to obtain the pectin-rich fraction at 10 MPa, 122 °C, a flow rate of 2 mL/min for 5 min. Water
was the control solvent for PLE and SWE methods. A semi-quantitative green metric tool, the
Green Certificate, measured the sustainability of the sequential extractions. The bioactive
potential of the anthocyanin-rich extract was evaluated by antioxidant, anti-glycemic, and anti-
lipolytic activity and thermostability. The pectin-rich extract was evaluated by water-holding
capacity (WHC), oil-holding capacity (OHC), and capacity to form and stabilize emulsions.
The anthocyanin fraction obtained using ChCl:Ma aqueous solution had a 40% higher yield
compared to water as solvent. Besides, the DES extract had antioxidant, anti-glycemic, and
anti-lipolytic activities 1-4 folds higher than the control extract, with less thermal sensibility
(1.2-folds higher activation energy, ) and good colorant stability, important characteristics for
a natural colorant. For the pectin fraction, the yield of the SWE modified by DES (Ca:Glu:Wa)
was 1.6-folds higher than the control, and with WHC and OHC values next to 1g/g, emulsifying
ability next to 50%, and emulsifying stability above 90%. The Green Certificate value for the
sequential extraction from jaboticaba peel was 90.35, a genuine green procedure, which
suggests its use for different biomasses. The presented sequential extractions contribute to a
biorefinery approach for the efficient use of natural resources, encouraging the use of DES in
high-pressure extraction methods to increase the value of agro-industrial by-products.

Keywords: Sequential extraction; deep eutectic solvent; green metric tools; pressurized liquid
extraction; subcritical water extraction.

57
Kappaphycus alvarezzi biorefinery: PLE and SFE applications for
phenolics and carragenan recovery and aerogel drying

Adenilson R. Rudke, Cristiano J. De Andrade, Sandra R. S. Ferreira*

Chemical and Food Engineering Department, Federal University of Santa Catarina, EQA/UFSC, ZIP
CODE 88040-900 Florianópolis-SC/Brazil

*s.ferreira@ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Kappaphycus alvarezzi (KA) is one of the world’s most cultivated (red) seaweed, very likely
due to the following main factors: rapid growth and high κ-carrageenan content. The
biopolymer κ-carrageenan is soluble in hot water and presents remarkable gelling properties,
with relevant uses in different industries (food, pharmaceuticals, and cosmetics). Carrageenan
is generally extracted by a two-step process: (I) pretreatment with KOH and (II) 2-4 hours hot
water extraction. Thus, it is essential to develop faster extraction techniques, reach similar or
higher yields, and maintain carrageenan properties. In this sense, the biorefinery concept can
be applied to KA biomass since it comprises of various compounds of interest. Therefore, this
work aimed to evaluate the Pressurized Liquid Extraction (PLE) with ethanol and water within
the biorefinery concept, to obtain phenolic compounds, carrageenan, and the cellulosic-rich
fraction. In addition, the recovered carrageenan was used for aerogel production and dried by
Supercritical Fluid Extraction (SFE). PLE, conducted at 100 bar and 40°C with ethanol, was
used to obtain the phenolic fraction. In contrast PLE at 50 bar and 80°C with water were used

58
to obtain the carrageenan, while the final solid material represented the rich cellulosic fraction.
The methods were compared to a conventional carrageenan extraction by maceration. Then,
yield, gel strength, and viscosity (carrageenan) were measured. The phenolics fraction from
KA, from PLE with ethanol, provided total phenolics content (TPC) of 7.82 ± 0.17, while the
Soxhlet method with ethanol reached a TPC of 3.42 ± 0.02. The carrageenan yield by PLE with
water was 62.64 ± 4.06 %, while the maceration method yielded 56.83 ± 1.90%. No statistical
difference was detected in the gel strength of the carrageenan fraction from PLE and maceration
methods. Otherwise, the carrageenan viscosity from PLE was slightly lower than the
conventional one, 46 ± 2 and 35 ± 1 cP, respectively. The SFE with CO2 at 40°C and 100 bar
was used for the ethanol removal from an alcogel formed from the recovered carrageenan,
producing an aerogel with similar characteristics, such as porosity and density, to that produced
by a commercial carrageenan. Therefore, the high-pressure methods PLE and SFE were
adequately applied to valorize KA biomass to recover valuable components for different
applications, such as aerogel production for food or pharmaceutical uses.

Keywords: Seaweed, sequential extraction, phenolics, polymer.

59
 Antioxidant extracts from olive pomace by compressed fluid
extraction technologies: comparison between SFE and PLE
Cecilia Daubera*, Melissa Romeroa, Adriana Gámbaroa, Elena Ibáñez b, Ignacio Vieiteza
a
Departamento de Ciencia y Tecnología de Alimentos, Facultad de Química, Universidad de la
República, Montevideo, Uruguay
b
Laboratory of Foodomics, Institute of Food Science Research, CSIC, Madrid,España

*cdauber@fq.edu.uy

GRAPHICAL ABSTRACT

ABSTRACT

Agroindustrial by-products represent a valuable source of bioactive compounds which may

be recovered and used for several applications like pharmaceutical, cosmetics or food
industry. Among them, olive pomace (OP) is a major problem for the olive industry due to
the high amounts generated during olive oil production and environmental concerns
regarding its phytotoxical effects on the soil. Bioactive compounds present in OP, mainly
polyphenols, are recognized to have positive effects on human health, related to their
antioxidant properties. In this sense, it is important to develop « green » extraction processes
that preserve the target compounds, minimizing the environmental load.
The aim of this work was to study and compare two technologies based on the use of
compressed fluids: supercritical fluid extraction (SFE) and pressurized liquid extraction
(PLE) to obtain natural extracts with antioxidant properties from the olive pomace.

60
OP of Arbequina variety was collected from a two-phase olive mill. A pre-treatment of
drying (40ºC, 22 h) and milling was performed before the extractions. Extractions for each
technology were carried out in a laboratory-scale equipment according to an experimental
design in order to know the effect of operational factors, in three response variables:
extraction yield (% w/w), total phenol content (TPC, expressed as mg GAE/g) and
antioxidant activity of the extracts (TEAC, expressed as µmol Trolox/g). For SFE, 15 runs
were carried out following a Box-Behnken Design in which the factors considered were:
extraction temperature (40-60ºC), extraction pressure (200-400 bar) and percentage of
ethanol as modifier (0-10%). For PLE, a face-centered Central Composite Design was
followed, comprising 11 runs in which extraction temperature (100-140ºC) and solvent
composition (50 - 100% ethanol in ethanol/water mixture) were the independent factors
considered. Process pressure remained constant for all runs at 10 MPa. Both designs included
three repetitions of the central point. TPC and antioxidant activity of the extracts were
determined by Folin-Ciocalteau and ABTS+. assays, respectively. Collected data for each
response variable was adjusted to quadratic regression models to determine optimal
extraction conditions.
For SFE, extraction yield was positively correlated with pressure and percentage of ethanol as
co-solvent, while TPC and TEAC were positively correlated with temperature and percentage
of ethanol as co-solvent. The higher values of TPC and TEAC were obtained at 20 Mpa, 60ºC
with 10% ethanol. For PLE, higher values of TPC and TEAC were obtained at 120ºC using
between 60 and 80% of ethanol, according to the quadratic model fitted to data (p<0.05).
When comparing the best OP extracts obtained by SFE (20 MPa, 60ºC and 10% ethanol as
co-solvent) and PLE (10 MPa, 120ºC, 75% ethanol) TPC and antioxidant activity was higher
for PLE extracts, as expected. Also, PLE resulted in higher extraction yields (20.4 vs 10.3 %
w/w), leading to a higher recovery of phenolic compounds (14.12 vs 0.13 mg GAE/g dw).
According to these results, PLE shows higher potential than SFE for recovering antioxidant
compounds from OP, which may be further used in food applications. In fact, the selected
PLE extract was able to protect sunflower oil from oxidative damage, increasing its induction
period 2.4 fold in an accelerated oxidative stability test (Rancimat) when added at 1000
mg/kg.

Keywords: Olive pomace, PLE, SFE, natural antioxidants

61
Effect of operational variables on the development of PLA/PBAT
active foams using supercritical fluids
Patricia Rivera a, b, Alejandra Torres*a, Julio Romero b, Francisco Rodríguez a, María José
Galotto a
a
Packaging Innovation Center (LABEN), Department of Food Science and Technology,
University of Santiago de Chile (USACH), Chile.
b
Laboratory of Membrane Separation Processes (LabProSeM), Department of Chemical
Engineering and Bioprocesses, University of Santiago de Chile (USACH), Chile.
*alejandra.torresm@usach.cl

GRAPHICAL ABSTRACT

ABSTRACT
In 2021, the total world production of plastics reached 391 million tons. Plastic materials are
massively used in various industries, mainly in food packaging, which generates large amounts
of polluting plastic waste. This has led to a great interest in the search for materials from
renewable sources to displace petroleum-derived polymers. Among the most interesting
biopolymers currently commercialized are cellulose, polylactic acid (PLA), starch, and
Polybutylene adipate-co-terephthalate (PBAT).
However, traditional methods of incorporating active agents into polymeric matrices have
some drawbacks, mainly due to their high volatility and thermosensitivity. An alternative to
this problem is the use of supercritical carbon dioxide (scCO2) due to its properties such as low
cost, non-toxic, environmentally friendly, and chemically inert. Moreover, scCO2 as a physical

62
blowing agent is advantageous because it expands rapidly due to a phase change at the blowing
temperature and pressure. Therefore, scCO2 foaming shows important advantages over the
traditional chemical foaming processes such as mass transfer properties adapted according to
the foam morphology, by generating different path lengths for the diffusion of the active
compounds within its structure.
The aim of this research was to study the effect of operational variables on the impregnation
and supercritical foaming processes of biocomposites with antioxidant activity based on
PLA/PBAT blends with caffeic acid (CA) using the supercritical fluid technique. The work
methodology considered the analysis of thermal (DSC and TGA) and structural (SEM and
FTIR) properties of the obtained materials. In addition, the antioxidant activity by DPPH assay
was evaluated.
The results revealed that the use of ethanol as a cosolvent contributed to an increase in the
amount of caffeic acid impregnated in the films. In addition, there was an increase in the degree
of crystallinity of the samples impregnated with CA, due to the nucleating effect of the active
compound. On the other hand, SEM images of both films and foams showed significant
differences due to the presence of PBAT and its low miscibility with PLA. Finally, the results
obtained in this work contribute to the knowledge of the important parameters to consider for
the implementation of the impregnation and foaming process of PLA/PBAT blends with
potential use in food packaging.
Keywords: PLA, PBAT, blend, active foaming, supercritical carbon dioxide.

63
 Supercritical CO2 Assisted Impregnation of Pink Pepper
Essential Oil in Gelatin-Siloxane Gels for Biomedical Applications
Ulisses Silva do Nascimentoa, Bruno Guzzo da Silvaa, Lígia Passos Maia-Obia*
a
Federal University of ABC, Avenida dos Estados, 5001, Santo André – SP, Brazil
*ligia.maia@ufabc.edu.br

GRAPHICAL ABSTRACT

ABSTRACT
Aside from its use as a condiment, pink pepper (Schinus terebinthifolius Raddi fruits) has high
commercial potential because of its wide range of biological activities, including anticancer,
anti-inflammatory, antioxidant, fungitoxic, antibacterial, and anti-Trypanosoma cruzi
activities. These properties have been related to the presence of terpenes from essential oil
(EO). Although these studies have shown that pink pepper is a promising source of bioactive
compounds, only a few studies have investigated impregnation and controlled release of the
pink pepper EO. The impregnation in polymers provides protective effect of the EO, preventing
losses of volatile compounds and the integrity of these compounds against degradation, and
enhancing biological activities through controlled release. Gels based on gelatin are
biocompatible and biodegradable materials that have called attention due its capacity to be used
for controlled release of actives and to absorb a significant amount of water, which is important
for its use as biomaterials. The aim of this work is to obtain two types of gels based on gelatin,
a xerogel and a cryogel, impregnated with pink pepper EO using supercritical CO2 and to verify
the impact of pressure and temperature at constant density on the EO load, and also to evaluate
the potential of the gels for biomaterial applications through their water absorption kinetics.
The gels were synthesized reacting gelatin with 3-glycidoxypropyl trimethoxysilane (GPTMS).

64
They were dried in oven to obtain a xerogel or liophilyzed to obtain a cryogel. FTIR showed
that the epoxide group (906 cm-1) from the GPTMS was no longer present in the sample
demonstrating that this group was connected to the gelatin chain and the bond Si-O-Si,
responsible for the crosslinking, was observed at 1086 cm-1. EO was obtained by steam
distillation in a pilot-scale distillation system. The EO was obtained with a yield of 6.0% ±
0.4% and the major compounds were ∆3-carene (37.21% - 40.35%), α-terpinene (18.91% -
19.54%), α-pinene (14.56% - 15.73), limonene (10.54% - 11.97%), and germacrene D (3.35%
- 4.23%). The impregnation was done at P = 100 bar, T = 35 °C, d = 0.71 g.ml -1 (condition 1)
and P = 200 bar, T = 60 °C, d = 0.72 g.ml-1 (condition 2) in a high pressure cell of 10 mL. Pink
pepper EO (10 %w/w in relation to the CO2) was added at the bottom of the cell and the samples
were allocated in stainless steel supports to prevent the direct contact with the pure EO. The
system was heated at the desired temperature and pressurized with CO2 using a manual syringe
pump from HiP and stirred for 4 h. The depressurization was made by cooling the cell with
liquid nitrogen before opening the exit valve, it took aprox. 10 minutes and its rate was not
controlled. The impregnation was evaluated gravimetrically, determining the %w/w of the EO
in the final material, and by FTIR. The potential for the material to be used as biomaterial was
investigated by the water absorption kinetics by immersing the samples in a pH 7.4 PBS buffer
solution at 37°C for 3 days, and their mass was measured in pre-determined time intervals. The
impregnation at the condition 1 showed loads of 0.2 %w/w for the xerogel and 4.9 %w/w for
the cryogel and at the condition 2, 0 %w/w for the xerogel and 7.2 %w/w for the cryogel. The
FTIR of the cryogels presented a clear increase of the C-H stretching band around 2900 cm-1
and a band at 1695 cm-1 due to C=C stretching indicating the presence of the alkenes that
composes the EO. The impregnation occurs for the cryogels, but is neglectable for the xerogels,
showing that the main mechanism of impregnation is related with the porous structure and not
with the diffusion of the EO into the matrix. Also, by increasing the pressure and the
temperature the load of the EO increases for the cryogel but decreases for the xerogel, it can
be explained by the fact that at higher pressures, the affinity of the EO by the CO2 increases,
and a greater amount of EO is carried into the pores and is entrapped by them when the system
is depressurized. However, by decreasing the pressure, the affinity of the EO by the CO2
decreases favouring the absorption equilibrium of the EO into the matrix (but it is absorption
is overall too low, indicating a low permeation of the CO2 into the matrix). Regarding the water
absorption, at the first minutes, it is greater for the cryogel, followed by similar absorption
speeds and maximum absorptions of 252 and 344 % for the cryogel and the xerogel,
respectively. This indicates that the materials are highly absorbent. The difference in absorption
is mainly related to their morphologies as the cryogel presents micropores that allow water
retention additionally to the swelling of the matrix where water occupies the interstices of the
chains network. It is possible to conclude that cryogels could be easily impregnated using
scCO2 and that further exploration of the operational conditions has potential to increase the
EO load. But the xerogel do not present the same potential. As these materials absorb a great
amount of water, the cryogel is shown to be promising as multifunctional material with
potential to be used for controlled release of the pink pepper EO for therapeutic or biomedical
applications. Additionally, this EO can be obtained by supercritical CO2 extraction, opening an
opportunity to integrate the processes, reducing energy costs.

Keywords: Pink pepper, essential oil, supercritical impregnation, gelatin xerogel, gelatin
cryogel

65
 Catalytic synthesis of ethyl levulinate aided by supercritical
carbon dioxide
Małgorzata E. Zakrzewskaa,*, Ana B. Paninhoa, Ana R.C. Duartea, Ana V.M. Nunesa
a
LAQV, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade
Nova de Lisboa, 2829 516 Caparica, Portugal
*corresponding author email address: ma.zakrzewska@campus.fct.unl.pt

GRAPHICAL ABSTRACT

ABSTRACT
Biofuels, understood as derived from biomass, are essential to meet fast-growing demand for
transportation and alleviate the negative impact of vehicles emissions. Alkyl levulinates can be
produced from levulinic acid, one of the top 12 platform chemicals produced from biomass,
and have been proposed as both gasoline and diesel additives capable of enhancing fuel quality
[1]. Ethyl levulinate is the most widely studied alkyl levulinate that is already being used in
flavoring and fragrance industry, plasticizers or as a solvent.
Conventionally, the Fischer esterification is carried out with strong mineral acid leading to
corrosion, environmental pollution, and troublesome catalyst recyclability [2]. On the other
hand, heterogenous catalyst are associated with lower reaction rates, side products or catalyst
deactivation. Ionic liquids, although emerged as a green solution, are often characterized by
complex preparation, possible toxicity, low biodegradability and high cost limiting their
applications to a large extent.
In this context, deep eutectic systems (DES) offer a pronounced advantage [3,4]. They are
recognized by their straightforward and reproducible preparation, 100% atom economy, low-
price, and often biodegradability and non-toxicity, especially in case of natural deep eutectic
systems (NADES) that are composed of primary metabolites like, i.e., organic acids, sugars,
amino acids or choline chloride.

66
In this work, we present the greener organic synthesis showing advantages of coupling dense
carbon dioxide with other environmentally friendly solvents such as eutectic systems. Catalytic
formation of fuel additives from a potentially biobased important platform chemical and
alcohol is investigated under various process conditions (i.e., temperature, pressure, time,
alcohol to acid ratio, or catalyst loading) in order to optimize the conversion and selectivity
towards the desired product. The obtained products are analyzed using GC coupled with FID
detector.

Acknowledgements
This project has received funding from the Fundação para a Ciência e Tecnologia FCT/MCTES
projects PTDC/EQU-EPQ/1039/2021 and UIDB/50006/2020 of the Associate Laboratory for
Green Chemistry – LAQV. This work was further supported by the European Union's Horizon
2020 (European Research Council) under grant agreement No ERC-2016-CoG 725034. GC-
FID and Mass Spectrometry data were obtained by the Laboratório de Análises/LAQV, and
the SCXR equipment was financed by national funds from FCT/MCTES - project RECI/BBB-
BEP/0124/2012.

References
1. J.F. Leal Silva, R. Grekin, A. Pinto Mariano, R. Maciel Filho, Energy Technol., 6 613
(2018).
2. E. Ahmad, Md. Imteyaz Alam, K.K. Pant, M. Ali Haider, Green Chem., 18 480 (2016).
3. A. Paiva, A.A. Matias, A.R.C. Duarte, Curr. Opin. Green Sustain. Chem., 11 81 (2018).
4. M. Komar, T.Gazivoda Kraljevič, I. jerkovič, M. Molnar, Molecules, 27 558 (2022).

Keywords: high-pressure carbon dioxide, eutectic mixture, organic synthesis

67
 Evaluation of the lipid fraction of tucumã-do-Amazonas
(Astrocaryum aculeatum) almond extracts obtained by SFE
Luciedry M. S. Carvalhoa*, Renato Grimaldib, Julian Martíneza
a
Laboratory of High Pressure in Food Engeering, Departament of Food Engineering and Technology,
Faculty of Food Engineering University of Campinas, UNICAMP, Campinas – SP, Brazil
b
Fats and oils Laboratory of, Departament of Food Engineering and Technology, Faculty of Food
Engineering University of Campinas, UNICAMP, Campinas – SP, Brazil
*corresponding author email address: eng.lmatheus@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT

The Amazon region has a very rich biodiversity in native species of great economic, academic
and social interests. Tucumã-do-Amazonas (Astrocaryum aculeatum) is a popular fruit from
the Amazon region, usually consumed as fresh, highly nutritious and a good source of oil that
can be extracted from its pulp and almond. The aim of this work was to extract the lipid fraction
from the almond of tucumã-do-Amazonas with supercritical CO2. Supercritical fluid extraction
(SFE) was performed at different conditions (15 MPa and 60 °C and 35 MPa and 40 °C) and
the extracts were characterized in terms of global yield, glyceride by high-performance size
exclusion chromatography (HPSEC), fatty acid and triacylglycerols profile (TAGs), using a
capillary gas chromatograph (CG), and thermal analyses by differential scanning calorimetry
(DSC). The global yield obtained for the evaluated conditions were 4.74 and 32.72% for 15
MPa and 60 °C and 35 MPa and 40 °C, respectively. The reason for this great difference in
global yield is that the solubility of the target compounds in supercritical CO2 increases with

68
density, which in turn increases with pressure, resulting in the enhanced recovery. Therefore,
the higher the pressure, the lower the solvent amount required for a specific extraction. The
increase in temperature decreases CO2 density, thus reducing target compounds' solubility in
this solvent. The lipid fractions extracted in both conditions are mainly composed of
triacylglycerols (TAG), with about 98%, and a minor amount of monoacylglycerols (MAG)
and free fatty acids (FA). Lauric (between 52,94 and 57,88%) and myristic (between 23,69 and
25,61%) acids, both saturated, were the most abundant fatty acids obtained in both conditions.
Regarding the the unsaturated fatty acids, oleic acid (between 4,14 and 5,85%) was the most
relevant. Triacylglycerols represent the largest class of glyceride compounds in the lipid
fractions of tucumã-do-Amazonas almond. The results show that the extracts are mostly
composed of TAG LaLaLa (between 24 and 28%), LaLaM (about 28%), and LaMM (between
12 and 15%) at the evaluated conditions evaluated. The thermal analyses showed that the
crystallization and melting curves of both extracts are very similar, which implies that they
were not affected by the SFE conditions at which they were obtained. Oils and fats do not have
specific melting and crystallization temperatures, but they have melting and crystallization
profiles. Based on the composition and thermal profile of the SFE extracts, tucumã-do-
Amazonas almond can be considered a good source of a lipid product, which is presented as a
fatty extract, obtained by SFE. It is also a good source of triacylglycerols, being lauric and
myristic the major fatty acids. The observed fatty acid profile, the TAG content and the thermal
profile of the extracts make them a potential ingredient for certain food formulations.

Keywords: SFE, food by-products, almond, lipid fraction, biodiversity

69
Supercritical extraction from Citrus reticulata Blanco with carbon
dioxide
Marcelle G. De F. Batistaa, Mônica Beatriz Kolicheskia, Fernando Augusto Pedersen Volla,
Marcos Lúcio Corazzaa,*
a
Department of Chemical Engineering, Federal University of Paraná, Av. Cel. Francisco H. Dos
Santos n. 100, Curitiba-PR, Brazil.
*corresponding author email address: corazza@ufpr.br

GRAPHICAL ABSTRACT

ABSTRACT

Production of tangerines and their hybrids in the world reached 55300 ktons/ year. Brazil
produced 984 ktons/year in an area of about 52000 hectares planted. Ponkan tangerines
belong to the Rutaceae family and have the scientific name of Citrus reticulata Blanco. The
fruits have an average of 162 g, 11 seeds, orange-yellow color, oblate shape, and rough skin
with prominent oil glands. The tangerine juice corresponds to 43 wt.% of the fruits and it is
normally extracted by pressing, generating an agro-industrial waste rich in compounds with
market value. Citrus peel has a high level of d-limonene, a terpene widely exploited in the
food, pharmaceutical, cosmetic and pesticide and cleaning product industries. Supercritical
fluid extraction is a technique in which volatile substances are obtained, mainly terpenoids,
and non-volatile substances such as oleoresins and fatty acids. Carbon dioxide is widely used
for extraction as a supercritical fluid due to its moderate critical pressure, 7.4 MPa, and low

70
critical temperature, 31 °C. Other advantage of using supercritical CO2 is its odorless and
colorless, it is safe, non-toxic and non-flammable. In the literature, there are studies with
supercritical CO2 of citrus fruits that obtained more than 95% d-limonene at conditions of 40
°C and at least 10 MPa. The objective of this study was to obtain an extract from Citrus
reticulata Blanco waste, rich in d-limonene. The Ponkan waste was purchased in the region
of Cerro Azul (Paraná, Brazil). This raw material consisted of peel, seeds and pulp and was
cut into pieces of approximately 1 cm², dried in an oven with air circulation at 50 °C for 15 h,
ground, and classified by sieving with particle size 28 mesh. The moisture content in dried
waste was 8.5 wt.%, determined in a Shimadzu infrared balance (model ID200). The essential
oil yield was 0.22 wt.% (88.5 % of d-limonene), it was determined in Clevenger aparatus for
2 h. The extraction yield using hexane was 1.94 wt.% (44.8 % of d-limonene), determined in
Soxhlet aparatus for 6 h. The extractions were performed by a supercritical extractor with
carbon dioxide as solvent. The static extraction time was 30 min and the dynamic extraction
used 20 g of biomass and flow rate of CO2 2 mL.min-1. The experimental planning of the
extractions evaluated two process variables: X1 - temperature (°C) and X2 - pressure (MPa)
at two levels (X1: 40 – 80 °C, X2: 100 – 200 MPa) and central point (X1: 60 °C, X2: 150
MPa), which factorial design type is 22. The extracts were analyzed by gas chromatography
mass spectroscopy (GC-MS), with a ZB-5MS column. The NIST (National Institute of
Standards and Technology) mass spectrometry library was adopted for the identification of
the other compounds present using Postrun GC-MS software. According to the literature,
pressure is the condition that most influences the extraction yield since the density and
solvation power of CO2 depend on the pressure. The influence of temperature depends on
each system, because increasing temperature increases vapor pressure and reduces densityn.
These effects were observed in this study: with the increase in pressure from 10 MPa to 20
MPa there was an increase in the extraction yield from 0.00 wt.% to 1.50 wt.% at 80 °C, and
from 0,6 wt.% to 1,29 wt.% at 40 °C. However, at fixed pressure of 20 MPa the extraction at
80 °C occurred at rate smaller than at 40 °C, but obtained an overall extraction yield greater,
1.50 wt.% and 1.29 wt.% respectively, which indicates that CO2 is acting only as a carrier of
the extract. The extracts obtained were analyzed by GC-MS and the highest fraction of d-
limonene obtained was 89%, at conditions 80 °C and 20 MPa. At 40 °C and 20 MPa the d-
limonene fraction obtained was 81.6%. The results show that the combination of both high
pressures and high temperature had a positive impact to recover extract rich in d-limonene,
which can be used in diferent industries.

Keywords: CO2, tangerin, extract, yield, d-limonene.

71
 INFLUENCE OF PHASE BEHAVIOR ON ENZYMATIC
ESTERIFICATION OF RESIDUAL OILS USING PROPANE
AS SOLVENT: EXPERIMENTAL STUDY AND
THERMODYNAMIC MODELING
Bruno M. Nogueiraa, Frederico W. Tavaresa,b , Papa M. Ndiayea,b,*
a
Institution 1: Programa de Engenharia Química/COPPE, Universidade Federal do Rio de Janeiro
(UFRJ)- Technology Center, Block G, Cidade Universitária, RJ 21941-914, Rio de Janeiro,
Brazil
b
Institution 2: Programa de Pós-Graduação em Engenharia de Processos Químicos e
Bioquímicos/EPQB, Escola de Química, Universidade Federal do Rio de Janeiro (UFRJ)-
Technology Center, Block E, Cidade Universitária, RJ 21941-909, Rio de Janeiro, Brazil
*corresponding author email address: papa@eq.ufrj.br

GRAPHICAL ABSTRACT

ABSTRACT

Enzymatic transesterification is an alternative promising route to produce biodiesel from

residual oil. The main characteristic of these oils is their high content of fatty acid. Usually, the
enzymatic transesterification is carried out using two steps. First, a hydrolysis of the
triglycerides is carried out to transform them into fatty acids and glycerol, then the resulting

72
fatty acid will be transformed into esters (biodiesel) through an esterification, both reactions
can be catalyzed by the free enzyme Eversa Transform 2.0 (ET2) from Novozymes. ET2
showed good performance in the presence of water with a significative reduction of its
denaturation, allowing its reuse. The major difficulty of these process is the time consuming
(more than 12h), for reaction and separation. The phase mass transfer plays an important role
mainly when two or more phases are presents. In this work propane is used as solvent in order
to optimize the enzymatic esterification reaction. Propane is known as a suitable solvent for
fatty acid and at high pressure, the contact of reactants molecule and ET2 can be significantly
improved. Additionally, when propane is used, the separation and purification process of
produced biodiesel is straightforward through pressure reduction. We investigate the phase
behavior of reactional mixtures with different propane molar fraction at several ration of
methanol to oil. The PC SAFT equation of state with suitable association scheme is used to
correlate experimental data.

Keywords: Enzymatic Esterification of Fatty acid, Phase behavior, propane, PC-SAFT

Modeling.

73
 Effect of supercritical CO2 extraction as pretreatment to obtain
c-phycocyanin from spirulina (Arthrospira maxima)
Juana A. López-Limóna, María A. Rios-Corripioa,, Gregorio Romero-de la Vegab*
Aleida S. Hernández-Cázaresa, Juan V. Hidalgo-Contrerasa, Ramzi del A. Mellado-Pumarinob
a
Institution 1: Colegio de Postgraduados, Córdoba, Veracruz , México
b
Institution 2: Universidad Iberoamericana Puebla-IDIT, Puebla, México
*corresponding author email address: gregorio.romero@iberopuebla.mx

GRAPHICAL ABSTRACT

ABSTRACT
C-phycocyanin (C-PC) is a blue pigment present in spirulina (Arthrospira maxima); it is a
water-soluble phycobiliprotein and photosynthetic accessory pigment. The main uses of C-PC
are as a nutritional use, flourescent marker or coloring in food and cosmetics [2].
Currently, Supercritical Fluid CO2
25
extraction (SFCO2) is being used as a
pretreatment alternative, having as its main 20
C-PC Yield (%)

function the denaturation of proteins; also

pretreatment with SFCO2 extraction has been 15
used to improve the antioxidant activity in
10
powder of spirulina [1,3]. The objective of
this work was to propose the SFCO2 5
extraction as pretreatment in spirulina
(Arthospira maxima) to facilitate the 0
A B C D
a b c d
posterior C-PC obtention from the raffinate. A: 75°C, 17.3 MPa +EtOH; B: 45°C, 24.2 MPa;
For the extraction, temperatures from 45 °C C: 75°C, 24.2 MPa + EtOH; D: 75°C, 31 MPa
to 75 °C and pressures from 17 to 31 MPa
were used, these conditions were performed Figure 1. Yield of freeze-dry powders.
using pure CO2 and 10% absolute ethanol for
120 min., subsequently the resulting raffinate were centrifuged adding distilled water at room

74
temperature at 5000 rpm for 30-40 min, the supernatant was freeze-dried to obtain the
𝑔 𝑜𝑓 𝐶−𝑃𝐶
characteristic blue powder of C-PC. The yield(𝑔 𝑜𝑓 𝑟𝑎𝑓𝑓𝑖𝑛𝑎𝑡𝑒 × 100) was then calculated and
expressed as percentage.
In the yields (Figure 1) it can be observed that the
best performance was the D treatment which was
achieved at the highest temperature and the
highest pressure without ethanol and the lowest
performance was at the highest temperature and
the lowest pressure with ethanol which
corresponds to C treatment (Figure 1); this can be
attributed to the temperature conditions to which
the sample has been subjected, since high
temperature denatures the proteins which results
in a low yield.
Figure 2. FTIR spectra of the freeze-dryThe freeze-dry powder underwent Fourier
powder with different treatments.
Transform infrared spectroscopy (FTIR)
analysis. Figure 2 shows the bands associated
with the presence of proteins in each of the samples also, a gradual absorbance increase is
observed in the bands, suggesting a protein concentration increase in the samples. Therefore it
can be confirm that the obtained blue powder contains C-PC.
These results allow to prove that the SFCO2 can serve as a pretreatment technology for the
spirulina conditioning for a subsequent C-PC extraction.
Keywords: c-phycocyanin, SFE, pretreatment, FTIR

References
[1] Dejsungkranont, M., Chen, H. H., & Sirisansaneeyakul, S. 2017. Enhancement of antioxidant
activity of C-phycocyanin of Spirulina powder treated with supercritical fluid carbon dioxide.
Agriculture and Natural Resources, 51(5), 347-354. DOI: https://doi.org/10.1016/j.anres.2017.12.001
[2] Eriksen, N. T. 2008. Production of phycocyanin—a pigment with applications in biology,
biotechnology, foods and medicine. Applied microbiology and biotechnology, 80, 1-14. DOI:
https://doi.org/10.1007/s00253-008-1542-y
[3] Olivera-Montenegro, L., Bugarin, A., Marzano, A., Best, I., Zabot, G. L., & Romero, H. 2022.
Production of Protein Hydrolysate from Quinoa (Chenopodium quinoa Willd.): Economic and
Experimental Evaluation of Two Pretreatments Using Supercritical Fluids’ Extraction and
Conventional Solvent Extraction. Foods, 11(7), 1015. DOI: https://doi.org/10.3390/foods11071015

75
 Extraction of neutral oil and polyphenolic fractions of spent
coffee grounds: single step and sequential extractions with
compressed fluids
Micheli Nolasco Araujoa, Natalia do Carmo Dinizb, Fabiane Hamerskib, Giuliana Varela
Garcia Lesakb, Júlio César de Carvalhoa and Marcos L. Corazzab*

a
Department of Bioprocess and Biotechnology Engineering, Federal University of Paraná, CEP
81531-990, Curitiba, PR, Brazil.
b
Department of Chemical Engineering, Federal University of Paraná, CEP 81531-990, Curitiba, PR,
Brazil.
* corazza@ufpr.br

GRAPHICAL ABSTRACT

ABSTRACT
In the climate crisis context, the replacement of traditional, land-using and food-competitive
raw materials with residues and by-products is imperative. The oils and polyphenolic fractions
of renewable matrices, such as coffee, are highly demanded by a wide range of industries such
as the pharmaceutical, cosmetics and food industries. Considering this, the use of spent coffee
grounds (SCG) instead of coffee beans for the extraction of the above-mentioned compounds
is a promising approach. The SCG extraction could be performed either in a single step or with
a sequential approach, in the latter of which a second extraction is carried out in the remaining
solid from the first step, allowing to further exploit the obtainable products. The sequential
extraction process alongside with other unit operations and raw materials is particularly
interesting in a biorefinery approach, since the remaining residue could be used either to
generate energy in boilers or second-generation biofuels, contributing to a circular economy.
In view of all of these aspects, this work assessed the single step and sequential extractions of
oil and polyphenolic compounds from spent coffee grounds. Furthermore, the compressed
propane and compressed butane, which application was not yet reported in the literature for
these purposes, were employed as the extraction solvents. Single step extractions were carried

76
out with pressurized propane and butane at different temperatures (40, 60 or 80 ˚C), pressures
(2, 6 or 10 MPa), static extraction times (0, 10, 30 and 60 min), dynamic extraction times (25
and 60 min), particle sizes (0.356 and 0.867 mm) and solvent mass flow rates (1, 2 and 3 g/min
for propane and 1 g/min for butane). Single step extractions were also performed with
supercritical CO2 with ethanol as the liquid solvent (scCO2+EtOH), with the following
experimental conditions of temperature, pressure and ethanol to SCG mass ratio: 80 ˚C, 20
MPa, 2 gethanol to 1 gSCG; 80 ˚C, 10 MPa, 2 gethanol to 1 gSCG; and 60 ˚C, 15 MPa, 0.5 gethanol to1
gSCG. Soxhlet extractions with organic solvents (n-hexane, ethyl acetate and ethanol) were also
carried out, and the compressed fluid extractions were compared with Soxhlet extraction with
n-hexane by means of the recovery parameter. Sequential extractions were performed as well,
in which the defatted solid resulting from the pressurized fluid extraction (with propane or
butane) was further extracted with supercritical CO2 with ethanol. The experimental conditions
of temperature, pressure and ethanol to defatted solid (DS) mass ratio were the same utilized
in the single step scCO2+EtOH extractions.
The extracts were analyzed by means of the fatty acids profile, total phenolic content,
differential scanning calorimetry and thermogravimetric analysis, and the resulting solid had
its soluble and insoluble lignin, structural carbohydrates and higher heating value quantified
by analytical methods. Two commercial green coffee oil samples were also analyzed and
compared with SCG oil samples.
The highest recovery among all single-step extractions (109.6 %) was obtained with Soxhlet
using ethanol. The extraction yields obtained with compressed propane and butane in different
operational conditions were statistically similar (average of 11.4 and 11.6 %, respectively),
indicating that mild operational conditions of pressure and temperature can be used without
yield penalty. When considering the highest overall sequential extraction yields, values up to
13.6 % and 13.1 % were obtained with compressed propane and butane, respectively, each one
at specific experimental conditions. Sequential extractions allowed a higher overall yield to be
obtained, since the single step extraction with scCO2+EtOH resulted in a maximum yield of
12.1 %. When compared to the single step scCO2+EtOH extract, the extract from the second
step of sequential extraction presented an increased (up to 431 %) phenolics content.
The similar yield results from propane and butane pressurized fluid extractions of SCG oil
preliminarily suggest the technical feasibility of the use of liquefied petroleum gas as the
extraction solvent, which is mainly a mixture of propane and butane and have a lower cost
when compared to the pure solvents. However, studies regarding the effect of the presence of
heavy contaminants in the oil should be performed to confirm this point. The results obtained
in this work showed that compressed propane and butane SCG oil extractions are promising
technical alternatives that can present advantages over conventional extraction methods such
as Soxhlet with n-hexane, such as shorter extraction times and easier solvent removal. The
exhausted solid, corresponding to 83 % of the original biomass, could be used to generate heat
in boilers, since its high heating value is only slightly lower from the SCG one. It also could
be used to synthesize second generation biofuels or chemicals due to its carbohydrate and lignin
contents. The similarity of the SCG oil obtained in this study with commercial green coffee oil
samples in terms of total phenolic content, fatty acids profile and thermal analyses results
suggested the possibility of substitution of coffee oil by SGC oil for some applications. The
results point to the technical feasibility of the extraction approach proposed in this study, that
allows the reutilization of the SCG, a widely produced residue worldwide, to produce added
valued products.
Keywords: Compressed propane, compressed butane, sequential extraction, supercritical CO2,
spent coffee grounds oil

77
 Interfacial description of binary systems containing CO2, n-
butanol and H2O.
Rafael Villablanca-Ahuesa,*, Roland Naglb, Tim Zeinerb, Philip Jaegera

a
Institute of Subsurface Energy Systems, Technische Universität Clausthal, Adolf Roemer-Straße 2A,
38678 Clausthal-Zellerfeld, Niedersachsen, Germany
b
Institute of Chemical Engineering and Environmental Techology, Technische Universität Graz,
Rechbauerstraße 12, 8010 Graz, Steiermark, Austria
*rafael.villablanca@tu-clausthal.de

GRAPHICAL ABSTRACT

ABSTRACT
Even though the most widely spread and studied area of thermodynamics in engineering
applications is that of the bulk phases, the thermodynamics of heterogeneous systems focusing
on their interface becomes of high relevance when processes need to be optimized and
simulated. For applications in the Oil and Gas industry, substances such as H2O, surfactants,
and gases such as CO2 are commonly present. Enrichment of surfactant-like components at the
interphases produces a change in the interface concentration with respect to the bulk phase,
which alters thermodynamic properties, potentially hindering diffusion as well as decreasing
the interfacial tension between coexisting phases affecting industrial processes such as
residence times in settlers and extraction.
The aim of the current work is to investigate the applicability of the Density Gradient Theory
(DGT) in combination with the Perturbed Chain Polar Statistical Association Fluid Theory
(PCP-SAFT) equation of state in order to describe interfacial thermodynamic behavior of
systems containing SC-CO2 in combination with a model surfactant (n-butanol) and H2O. The

78
vapor-liquid systems n-butanol+CO2 and H2O+CO2, as well the liquid-liquid and vapor-liquid
subsystems formed by CO2+n-butanol+H2O were studied at saturation conditions. In particular,
the phase density and interfacial tension of the systems were measured by the Oscillating tube
technique and Pendant Drop method, respectively, between 313.15 and 323.15 K up to 30 MPa.
These properties aforementioned, as well as phase equilibria were theoretically estimated by
the DGT+PCP-SAFT approach.

Keywords : Interfacial Tension, Surface active, PCP-SAFT, DGT, Experimental.

79
 Extraction of volatile oils from aromatic Ecuadorian plants
Lorena I. Jaramilloa,b*, Edwin Veraa, Jalloul Boujilab, Séverine Camyb
a
Escuela Politécnica Nacional, Ladrón de Guevara17-12-866, Quito, Ecuador
b
Université de Toulouse, INPT, UPS, Laboratoire de Génie Chimique, UMR CNRS 5503, 4 Allée
Emile Monso, F-31030 Toulouse, France
*corresponding author email address: lorena.jaramillo@epn.edu.ec

GRAPHICAL ABSTRACT

ABSTRACT
Ecuador owns a great biodiversity thanks to its different climates and geographical location.
Sunfo (Clinopodium nubigenum) and Muña (Minthostachys mollis) are aromatic plants
belonging to the Lamiaceae family, which have been used by the autochthones for centuries
against gastritis, anthi-influenza, menstrual symptoms, etc. Essential oils (EO), oleoresins (OR)
and volatile oils (VO) are mixtures of hydrocarbons, aldehydes, alcohols, and phenols, which
confer them among others antibacterial, antifungal, antioxidants, antitumor properties.
Therefore, they have gained importance in the food and pharmaceutical industry.
Different techniques have been used to obtain extracts (EO, OR) from plants such as
hydrodistillation (HD) and soxhlet (SX), these techniques have disadvantages such as long
extraction times, use of high temperatures or solvents. Alternatively, there are unconventional
extraction techniques like ultrasound assisted extraction (UAE) and supercritical fluid
extraction (SFE), which are performed at low temperature, reduce energy consumption, and
enhance the extraction yield. SFE is a green technology that uses supercritical CO2 as a solvent
and provides extracts that are free of solvent.
There are no studies of VO extraction by SFE from Sunfo and Muña. Therefore, this work aims
to study the influence of the extraction process conditions on the extraction yield of oils from
Sunfo and Muña obtained by SFE and compare it with the one obtained by HD, SX and UAE.

80
Experiments were performed to determine the pressure (100-600 bar), particle size (0.35-1.0
mm), CO2 flow (10 -20 g/min), temperature (40 -60°C) and static time (10 – 20 min) allowing
to obtain the highest global yield. The identification and quantification of the major compounds
was done by means of gas chromatography coupled to mass spectrometer. Finally, preliminary
experiments were performed to analyze the inhibition growth of two fungi (M. racemosus, M.
racemosus) that affect Ecuadorian flora by means of plate dilution at different extract
concentrations (400 and 800 μg/ml).
The pressure has a significant positive influence on the yield until 300 bar, particle size has an
inverse influence, whereas solvent flow and static time do not have influence on the yield. The
extraction yield varied according to the extraction technique for Sunfo and Muña as follows:
SX (258 / 299 mg/g), UAE (227 / 241 mg/g), SFE (36 / 21 mg/g) and HD (27 / 7 mg/g).
The composition of extracts for both plants varied strongly depending on the extraction
method. The major compounds of Sunfo were menthol, pulegone, thymol, carvacrol,
caryophyllene and piperitenone, and, on the other hand, the major compounds for Muña were
humelene, piperitone and caryophyllene. Finally, the extracts from both plants inhibited
partially or entirely microbial growth. The highest inhibition was reached by VO´s Muña
(96.27 ±1.00 %) for M. racemosus. On the other hand, VO from Muña and Sunfo inhibited
completely C. musae´s growth. These results confirm the potential of Minthostachys mollis and
Clinopodium nubigenum as a promising natural antifungal agent thanks to their chemical
composition.

Keywords: Please include at the end three to five keywords separated by a comma (Times New
Roman 12, single space).

81
 Nutraceutical delivery via SC-CO2: Lentil protein and
pectin based aerogels to encapsulate β-carotene
Srujana Mekalaa,, Marleny D.A. Saldaña
a
University of Alberta, Edmonton, AB, Canada
*corresponding author email address: marleny@ualberta.ca

GRAPHICAL ABSTRACT

ABSTRACT
β-Carotene is a naturally occurring pigment that also functions as an antioxidant with various
health benefits. Due to poor stability of β-carotene against temperature, oxygen, and light,
incorporating it into delivery systems enhances its bioavailability and application in the food
and beverage industry. Lentil protein and pectin are gaining popularity as wall materials for
delivery systems due to their abundance, biocompatibility, and functionality. In this study, we
developed β-carotene loaded aerogels with lentil protein and pectin as wall materials using
supercritical CO2 drying. The aerogels were formed in two steps: 1) formation of β-carotene
loaded emulsion gels using high-intensity ultrasound (HIUS), 2) emulsion gels were then
converted to alcogels via solvent exchange and then dried using SC-CO2 to form aerogels. The
physico-chemical, mechanical, and functional properties of aerogels were determined using

82
various analytical techniques. The structural conformation studies of the aerogels revealed the
predominance of hydrogen bonding responsible for the interactions between the lentil protein
and pectin. The water absorption capacity of the aerogels were in the range of 2.1 – 9.7 g
water/g aerogel while the oil absorption capacity was in the range of 3.7 – 9.4 g oil/g aerogel.
The release kinetics and antioxidant activity of the aerogels were also studied. Overall, the β-
carotene loaded aerogels using lentil protein and pectin by SC-CO2 drying provided a novel
approach for promoting stability and delivery of β-carotene. Utilization of naturally occurring
biopolymers lentil protein and pectin offer environmentally begin and sustainable solutions.
The aerogels developed in this study have potential applications in food industry.

Keywords: β-Carotene, aerogel, lentil protein, SC-CO2 drying, high-intensity ultrasound

83
 Ionic gelation assisted by compressed fluids (IGACF):
a new process for microencapsulation of bioactives in chitosan
Camilo Pardo-Castaño and Gustavo Bolaños*

Laboratory of Applied Thermodynamics and Supercritical Fluids

School of Chemical Engineering, Universidad del Valle, CL 13 100-00 Edificio E30, Cali, Colombia.
*corresponding author gustavo.bolanos@correounivalle.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
The use of chitosan-based materials for micro- and nanoencapsulation of pharmaceutically
active compounds has been growing during the last years, and several techniques have been
widely used for this purpose, but they usually have important drawbacks such as thermal
degradation of the product, poor control of the particle size distribution and use of organic
solvents. Supercritical fluid assisted processes, such as SAA (Supercritical Assisted
Atomization), RESS (Rapid Expansion of Supercritical Solutions), SAS (Supercritical
AntiSolvent), and SCASA (sc-CO2 Solubilization and Atomization) have been overcoming
such drawbacks. However, long solubilization times, high operating pressures and high drying
temperatures are still a matter of concern to improve processing efficiency, operating costs,
and quality of the product. In this work, we propose a new process we designate as Ionic
Gelation Assisted by Compressed Fluids (IGACF) to microencapsulate bioactive molecules in
chitosan, and we illustrate the process characteristics by producing microparticles of two

84
biologically active derivatives of coumarin, using commercial chitosan as encapsulating
material.
Two highly bioactive isomers referred as collinin and isocollinin were synthesized in a previous
work and were characterized by differential scanning calorimetry (DSC), infrared spectroscopy
(FTIR) and nuclear magnetic resonance (NMR). These compounds exhibit strong activity
against periodontal and other common persistent bacteria. A batch of commercial chitosan was
purified to obtain a 17% acetylated product with an average molecular weight of 340 kDa,
which was used as the encapsulating material.
In IGACF, the active compound and chitosan are initially suspended in water. In the presence
of compressed carbon dioxide chitosan is dissolved, and pentasodium triphosphate (TPP)
dissolved in water is slowly injected. TPP is used as a crosslinking agent, to induce ionic
gelation at 30 ºC and 70 bar in 3 h and to form solid aggregates (i.e., bioactives-in-chitosan
microparticles), which can be separated by centrifugation or freeze drying.
Initially, a 23 factorial experiment was run without the presence of the bioactive compound to
determine the effect of the mass concentration of chitosan (2 to 4 wt%), the mass concentration
of TPP (1 to 2 wt%), and the mass ratio of chitosan/TPP (2.0 to 3.1) on the microparticles
morphology and size distribution. The results show that particles with different morphologies
(spherical and amorphous) are obtained, with average particle sizes from 0.2 to 6.4 µm and
polydispersity indexes of up to 0.01. When similar experiments were run for including a load
of collinins, the obtained particles have 2.9 µm average size, 0.05 polydispersity index and 4%
loading content, with a 72% entrapment efficiency. All obtained microparticles were
characterized by FTIR, DSC, and electron microscopy (SEM-EDS).
This process represents a promising and ecofriendly alternative for producing microparticles
of bioactive compounds encapsulated in chitosan. These microparticles are useful in the
pharmaceutical industry to produce controlled release products. The new process uses low
pressures (up to 70 bar) and temperatures (up to 50 ºC), in short operating times (up to 3 hours),
with no use of any organic solvent.

Keywords: chitosan, collinin, microparticles, ionic gelation, carbon dioxide.

85
CO2 + Ethanol Assisted Impregnation of Chitosan Microparticles
with Curcumin
Pedro H. F. Tondo, Rubem M. F. Vargas*, Eduardo Cassel

Unit Operations Lab, School of Technology, PUCRS, Ipiranga, 6681, Porto Alegre, Brazil
*rvargas@pucrs.br

GRAPHICAL ABSTRACT

ABSTRACT
Curcumin, a polyphenol extracted from turmeric, has many therapeutic properties. Those
benefits make this compound a promising alternative in medicine that can be used in the
treatment of diseases such as arthritis, metabolic syndrome, liver disease, obesity,
neurodegenerative illness and multiple types of cancer. Colon cancer is high occurrence disease
and it’s the second most lethal type of cancer. As chemotherapy has too many side effects, new
natural products are widely researched as alternative drugs to treat all kinds of cancer.
Curcumin is being researched as a replacement for the traditional chemotherapy drugs,
however, it’s instability when ingested makes it unable to reach the colon with treatment
potential. Incorporating the compound in a biopolymer is a common strategy to by-pass this
problem and allow the drug to reach the desired location. Chitosan is an example of biopolymer
which suits these characteristics and can be used to transport the curcumin through the digestive
tract. Supercritical impregnation was the technique chosen to incorporate the curcumin into the
chitosan, which is a new method that allows a more practical control of the final properties of

86
the polymer. The goal of this work was to impregnate the chitosan particles (100 mg) with
curcumin (20 mg) using carbon dioxide expanded in ethanol and evaluate its delivery capacity
as well as making the mathematical modeling of the process. A 23 factorial design was carried
out to evaluate which condition of co-solvent percentage (50%, 60%, and 70%), and pressure
(100, and 200 bar), at 55 °C and depressurization rate of 10 bar.min-1, resulted in a higher
percentage of curcumin impregnation in the chitosan. SEM and FTIR were used to characterize
the particles before and after the impregnation. The pressure of 200 bar resulted in highest
percent and amount of curcumin impregnated, 15.8% and 3.16 mg, in the particles and there
was no significant difference between the quantities of ethanol evaluated. The first order model
was the one that best described the curcumin liberation in PBS (pH 7.4). Furthermore, these
experiments indicated that the microparticles released concentrations between 20 and 48 µM
of the active compound in the first 120 min.

Keywords: colon cancer, drug delivery, mathematical modeling, co-solvent, SC-CO2

87
 Comparison of emerging extraction techniques for recovering
phenolic compounds from avocado byproducts using a Natural
Deep Eutectic Solvent based on chloride choline and lactic acid
Juan F. Grisales-Mejíaa,*, Valeria Cedeño-Fierroa, Jenny Pala Ortegab, Harlen G. Torres-
Castañedaa, Margarita M. Andrade-Mahechaa, Hugo A. Martínez-Correaa,*, Gerardo Álvarez-
Riverac, Jose A. Mendiolac, Alejandro Cifuentesc, Elena Ibañezc
a
Universidad Nacional de Colombia, Carrera 32 # 12 – 00, Palmira, Valle del Cauca, Colombia
b
Universidad Nacional de Colombia, Carrera 30 # 45 – 03, Bogotá, Colombia
c
Institute of Food Science Research - CIAL (CSIC-UAM), Nicolas Cabrera 9, 28049 Madrid, Spain
*corresponding authors email address: jfgrisalesm@unal.edu.co & hmartinezco@unal.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
Phenolic compounds have gained significant interest in industries developing consumer
products due to their beneficial biological properties for health, particularly their well-known
antioxidant properties. In pursuit of a more sustainable economy and aligned with the
Sustainable Development Goals (SDGs), emerging extraction technologies have been
implemented to obtain these compounds. When combined with green solvents, such as Natural
Deep Eutectic Solvents (NADES), they become environmentally friendly alternatives to
conventional methods that are known to be harmful to human health and the environment.
However, there have been limited studies integrating these solvents with extraction methods,
particularly those involving high pressures. Thus, the objective of the present investigation was
to compare the influence of Pressurized Liquid Extraction (PLE) and Ultrasound-Assisted
Extraction (UAE) using a NADES based on choline chloride and lactic acid (ChCl:Lac) on the
phenolic compounds found in Hass avocado peels and seeds. These residues were dehydrated
in a freeze dryer (-40 °C, 120 mbar) and reduced in size using a blade mill to obtain a diameter
between 0.2 and 0.6 mm. The ChCl:Lac synthesis involved mixing the individual compounds
in a 1:1 molar ratio and heating them in a water bath at 85 °C until a transparent liquid was

88
formed. To aid the formation of the NADES, 10% of the total water to be used (50% of the
total solution) was added. The UAE technique was performed in an ultrasonic bath at 40 kHz
and room temperature for 30 minutes, while PLE was conducted at 100 bar, 100 °C, with 20
minutes of static time. The in vitro assay results for total phenolic content (TPC) showed that
the epicarp extracts obtained through PLE (166 ± 8 mg gallic acid equivalent [GAE]/g dry
sample [DS]) had higher values compared to UAE (148 ± 1 mg GAE/g DS), likely due to the
conditions employed, particularly the temperature. Unsignificant differences were observed in
the case of the seed extracts (56 ± 3 and 58 ± 1 mg GAE/g DS, respectively). One possible
explanation for this relates to the structural changes mediated by the interaction between seed-
starch, water, and NADES at that temperature, which may have limited further TPC extraction.
Subsequent chemical characterization of the extracts using HPLC-ESI-QTOF-MS revealed
that the epicarp extracts obtained by PLE contained a greater number of phenolic compounds
(33) compared to UAE (27), particularly phenolic acids, as well as other types of compounds
including some fatty acids. In contrast to the TPC assay, PLE demonstrated superiority in the
case of the seed extracts, with a total of 36 compounds compared to the 26 tentatively identified
in the UAE extracts, with the main difference being the presence of proanthocyanidins. These
results demonstrate that PLE is an emerging tool that can contribute to the bioeconomy by
enabling the production of high-value bioproducts from plant waste matrices, further enhanced
by the use of innovative solvents like NADES.

Keywords: Green chemistry, neoteric solvents, waste, subcritical water, Persea americana
Mill.

89
 EXTRACTION AND EVALUATION OF PEACH (Prunus
persica) SEED OIL OBTAINED BY SUPERCRITICAL CO2
EXTRACTION
Fredy Huaytaa*, Harold Pajanb , Henry Obregóna, Joel Sedanob, Jean Paul Rosalesb, José
Arguméb, Jeremy Palaciosb
a
Pontificia Universidad Católica del Perú, Av. Universitaria 1801, San Miguel, Perú
b
Universidad Nacional de Ingeniería, Av. Túpac Amaru 210 – Rímac, Lima, Perú
* corresponding author email address: fhuayta@pucp.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
The use of carbon dioxide (CO2) as a solvent in the supercritical fluid extraction process has
been successfully applied in the extraction of oils from various seeds, being considered a
benign technology for the environment. Therefore, in this research, a study was carried out to
optimize the yield in the extraction of peach seed oil (Prunus persica) at the National University
of Engineering in the Unit Operations Laboratory using the Applied Separations basic model
Helix SFE System supercritical fluid equipment. The company Proexi SAC (Peru-Lima-San
Isidro) donated 17 Kg of peach pit (seed plus stone or bone or endocarp), the seed represents
13.92% of the pit and has 4.76% moisture. The seed was crushed to a particle size between 850

90
and 1180 µm. In the extraction process, temperature, pressure, and CO2 flow were considered
as study variables. The Response Surface Methodology was used to determine the optimal
values of pressure and temperature. The results show that operating at 2 liters per minute of
CO2, a yield of 16.5% is obtained experimentally at 350 bar and 50 °C and a yield of 17.3 % is
estimated under conditions of 331 bar and 35.86 °C. The two types of volatile components of
peach seed oil that have the highest relative area are aldehydes and alcohols such as
Benzaldehyde (41.37%, aldehyde); 2,4-Decadienal, (E, E) (18.03%, aldehyde) and Benzyl
alcohol (15.27%, alcohol), also in smaller quantities there are acids (7.36% Oxalic acid),
ketones (5.15% - 3-Hexanone, 2-Hexanone) and phenols (4.02% - Butylated Hydroxytoluene).
These results show that peach seed moisture favors food preservation and is an oil enriched
with volatile compounds having a characteristic peach aroma which can be very useful for the
cosmetic and food industry.

Keywords: Extraction, peach seed oil, volatile compounds, optimization parameters,

supercritical fluid

91
 Supercritical CO₂ Extraction Of Oil From Soldier Fly Larva
MeaL (Hermetia illucens L) – Yield And Antioxidant Activity
Vanessa A. Cruz a, Alessandra L. De Oliveira a*

Faculty of Animal Science and Food Engineering, University of São Paulo, Av. Duque de
Caxias Norte, 225, Pirassununga, São Paulo, CEP: 13.635-900, Brazil
*Corresponding author: alelopes@usp.br

GRAPHICAL ABSTRACT

,,

ABSTRACT
With the growth of the world population and for food, edible insects have been drawing
attention for their economic, nutritional and sustainable benefits (Hierro, et al., 2020). The black
soldier fly (Hermetia illucens) has stood out for its nutritional composition, as it contains a high
content of proteins, lipids and antioxidants (WANG et al., 2017). Extraction with supercritical
CO₂ (scCO2) was applied because it has many advantages, such as not leaving toxic residues
in the flour or in the extracted oil. This research studied the profile of antioxidants present in
black soldier fly oils in different dynamic and intermittent processes.
Extractions with scCO2 were performed at a temperature of 60°C and pressures of 25 and 30
MPa. Static time (tE) of 30 min and continuous flow (10 mL/min) for all experiments.
The intermittent extraction process was carried out in 3 cycles. Starting the first cycle (C), in
which the flour with the solvent remained in contact for 30 min of static time (St), after this

92
period the pump was turned on and the outlet valve opened to collect the oil until reaching the
mass of determined CO₂. Once the collection was complete, a new cycle began with the same
contact time and P and T. In the dynamic process, the extraction was performed in 90 min. The
analyzed parameters were the yields and the antioxidant profile determined by the 2,2'-azinobis
(3-ethylbenzothiazoline)-6-sulfonic acid radical cation (ABTS) and ferric reducing antioxidant
power (FRAP) according to RUFINO et al., 2010 .
The dynamic and intermittent processes showed similar yields of 33% of oil, which corresponds
to 32.70% of the oil contained in the flour, not differentiating between them. The intermittent
showed lower consumption of CO₂, with half the volume of solvent spent in the dynamic
process. Intermittent extraction showed the same yield as dynamic extraction. However,
extraction time and solvent consumption were considerably lower. It is believed that this
extraction method in cycles (intermittent process) should be adopted, due to its important
economic benefits.
The antioxidant capacity of black soldier fly oil 0.23 ± 0.13 to 0.50 ± 0.11 µmol TE/g of oil did
not show statistical differences between them, however, the extracts obtained under conditions
of higher pressure showed higher concentrations of antioxidant activity. In the FRAP test, the
reducing antioxidant power also did not differ from each other (5.44 ± 0.89 to 6.59 ± 0.99 mg/g)
showing a lower oxidative potential when compared to Silver Spur apple peel (50.47 ± 7.22),
indicating that fly oil has low oxidative potential.
It was possible to conclude that the extraction method in cycles (intermittent process) should
be adopted, due to its yield and economically important benefits, since the solvent consumption
(scCO2) is lower. Black soldier fly oil showed potential as a product with antioxidant activity,
being an important factor for human nutrition or even for its use in cosmetics formulation.

References
Navarro Del Hierro, J., Gutiérrez-Docio, A., Otero, P., Reglero, G., Martin,
D. Characterization, antioxidant activity, and inhibitory effect on pancreatic lipase of extracts
from the edible insects Acheta domesticus and Tenebrio molitor. Food
Chemistry, 309, 2020. https://doi.org/10.1016/j.foodchem.2019.125742
Rufino, M. Do S. M.; Alves, R. E.; Brito, E. S.; Perez-Jimenez, J.; Saura-Calixto, F.; Mancini-
Filho, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits
from Brazil. Food Chemistry , v.121(4), p.996–1002. 2010.
Sethi, S., Joshi, A., Arora, B., Bhowmik, A., Sharma, Rr E Kumar, P. Significado dos ensaios
FRAP, DPPH e CUPRAC para determinação da atividade antioxidante em extratos de
maçã. European Food Research and Technology , 246 , 591-598. 2020.
Wang, C. Liang.; Wang, W.; Wang, T.; Deng, Z.; Yang, F.; Xiong, J.; Feng, W. Exploring the
potential of lipids from black soldier fly: New paradigm for biodiesel production (I).
Renewable Energy , v. 111, p. 749–756, 2017. Disponível em:
<http://dx.doi.org/10.1016/j.renene.2017.04.063>.

93
 New Deep Eutectic Solvents based on hyperbranched
polyglycerols and superbases for CO2 absorption
Gabriel Silveira dos S.*, Diego Miranda de S. C., Reinaldo C. Bazito

Fundamental Chemistry Department, Institute of Chemistry University of São Paulo

Av. Prof. Lineu Prestes, nº 748, São Paulo, Brasil
*gss.silveira97@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
High emissions of gases such as carbon dioxide (CO2) intensify the greenhouse effect and
impact the average temperature of planet Earth. These temperature variations can cause
ecological imbalances, climate changes, and other phenomena that harm the environment. In
light of this, the scientific community has been developing various strategies to efficiently
capture CO2 without causing harm to the environment. Deep eutectic solvents (DES) are
environmentally friendly materials for CO2 absorption compared to other known absorption
methods, such as traditional aqueous solutions of ethanolamine and its derivatives, and
applications with ionic liquids.
DES are liquid solvents at room temperature. They are composed of a mixture of two or more
substances in which the molecules interact with each other through hydrogen bonds, resulting
in a significant lowering of the mixture's melting point. The precursors of DES can be simple,
low-cost substances that are generally less toxic to the environment. The lowering of the
melting point occurs through the interaction between a hydrogen bond donor (HBD) molecule,
such as alcohols, amines, and carboxylic acids, and a hydrogen bond acceptor (HBA) molecule,
such as organic salts like choline chloride (ChCl). To increase the DES's affinity for CO 2, other
substances can be added to the mixture, and HBD or HBA can be functionalized. Superbases,

94
in general, are known for their affinity with CO2 and have been used in DES formulations from
simpler molecules. Larger molecules, such as hyperbranched polyglycerols (PGOH), can be
used as HBD due to the high density of functional groups in the polymer chain. PGOH has
multiple hydroxyl groups along the chain and is a good candidate for DES synthesis.
This work aims to present the results of DES synthesis using low-molecular-weight
hyperbranched polyglycerols as the hydrogen donors and choline chloride as the hydrogen
acceptor, as well as other organic salts. Ternary mixtures containing superbases will also be
presented to evaluate CO2 formation. The base used will be 1,4-diazabicyclo[2-2-2]octane
(DABCO).
The DES were characterized by nuclear magnetic resonance (NMR) of carbon and hydrogen,
matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF),
and thermal analyses such as differential scanning calorimetry (DSC). CO 2 absorption analyses
of the different mixtures were performed on a magnetic suspension balance.

Acknowledgments : We gratefully acknowledge the support of the RCGI – Research Centre

for Greenhouse Gas Innovation, hosted by the University of São Paulo (USP) and sponsored
by FAPESP – São Paulo Research Foundation (2020/15230-5; 2014/50279-4) and Shell Brasil,
and the strategic importance of the support given by ANP (Brazil’s National Oil, Natural Gas
and Biofuels Agency) through the R&DI levy regulation.

Keywords: CO 2 absorption, deep eutectic solvents, superbase, hyperbranched polyglycerol.

95
 Chitosan-TPP based aerogels for tissue engineering applications
Cristiana S. A. Bento, José P. Ruivo, Hermínio C. de Sousa,
Ana M. A. Dias*, Mara E. M. Braga*

Chemical Process Engineering and Forest Products Research Centre (CIEPQPF), Department of
Chemical Engineering, University of Coimbra, 3030-790 Coimbra, Portugal
*adias@eq.uc.pt; marabraga@eq.uc.pt

GRAPHICAL ABSTRACT

ABSTRACT
Tissue engineering scaffolds have been a topic of research in the last decades aiming the
development of scaffolds that can promote tissue repair when self-regeneration is compromised
[1,2]. Aerogels present ideal properties for this purpose, such as high and tailorable porosity
and tunable surface chemistry and bulk properties [3]. There has been an effort to produce eco-
friendlier and biocompatible aerogels by processing different biopolymers. Among others,
chitosan (CS) is one of the most studied for application as scaffolds for tissue engineering
applications, including as wound dressings, especially due to its biocompatibility,
biodegradability, and hemostatic and antimicrobial activities.
CS-based structures can be obtained by ionic and/or covalent crosslinking. Sodium
tripolyphosphate (TPP) is an ionic crosslinker with low toxicity that has been intensively
employed for the production of chitosan-based nanoparticles, hydrogels and scaffolds [1]. CS-
TPP cryogels were already reported in the literature [1;4] however, to our knowledge, there is
no report on the use of supercritical CO2 (scCO2) drying to obtain CS-TPP aerogels. Among
other advantages, scCO2 drying allows the production of sterilized/disinfected scaffolds with
high and tunable surface area and porosity while preserving the hydrogels' chemical and
morphological properties, which are critical properties when biomedical applications are
envisaged [3,5].
In this work, biocompatible and biodegradable CS-TPP aerogels were developed using scCO2
drying. Tridimensional CS-based hydrogel structures were obtained by ionic crosslinking of
low molecular weight chitosan (deacetylation > 75%) with TPP at different concentrations (0.5;
1.5 and 2.5% w/v). CS (3% w/v) was dissolved in an acetic acid solution (1% v/v) under stirring
for 24 h. A known volume of CS solution ( 20 ml) was then poured into a Teflon mold which

96
was further immersed in the TPP solutions. A composite polymer network was also prepared
by mixing CS (at 3% m/v) with poly(vinyl alcohol) (PVA, Mw 89,000-98,000, 99+%
hydrolyzed) at 1% (m/v) to evaluate its effect on the mechanical properties of the CS-TPP
aerogels. The mixture was further crosslinked with TPP at 1.5% w/v (intermediate TPP
concentration tested). All the prepared hydrogels were washed with water to remove non-
crosslinked TPP. The solvent exchange at ambient temperature was performed by sequential
immersion of the gels into ethanol/water mixtures, followed by scCO2 drying (2h, 150 bar,
35ºC, 0.5 L.min-1). The obtained aerogels were fully characterized in terms of their
physicochemical, morphological and mechanical properties by Fourier transform infrared
spectroscopy (FTIR), water swelling capacity (when immersed in PBS at 25ºC), water vapour
sorption and transmission rate capacities, helium pycnometry, nitrogen adsorption, optical
microscopy and compressive strength.
Results show that all the produced CS-TPP aerogels are mesoporous (pore diameters  12 nm)
and that their surface area increases with the crosslinking degree (ranging between 150 and 190
m2/g). Moreover, compressive tests allowed concluding that higher crosslinked CS-TPP
aerogels present higher mechanical strength. The average water vapor transmission rate of the
aerogels was  1550 g.d-1.m-2, although a lower value (~ 1260 g.d-1.m-2) was observed for CS-
TPP2.5%.
This work shows that it is possible to tune several properties of CS-TPP-based aerogels, such
as porosity, surface area, water swelling capacity or mechanical properties, by using non-toxic
and simple procedures, targeting different tissue engineering applications including wound
dressing biomaterials.

Keywords: cross-linked chitosan; sodium tripolyphosphate (TPP), supercritical drying

(scCO2); aerogels

REFERENCES
[1] I. Silvestro, I. Francolini, V. Di Lisio, A. Martinelli, L. Pietrelli, A. Scotto, A. Scoppio, A. Piozzi,
Preparation and Characterization of TPP-Chitosan Crosslinked Scaffolds for Tissue Engineering,
Materials, 13 (2020) 3577.
https://doi.org/10.3390/ma13163577
[2] D. Izzo, B. Palazzo, F. Scalera, F. Gullotta, S. Scialla, A. Sannino, F. Gervaso, D. Izzo, B. Palazzo,
F. Scalera, F. Gullotta, Chitosan scaffolds for cartilage regeneration : influence of different ionic
crosslinkers on biomaterial properties, Int. J. Polym. Mater. Polym. Biomater. 68 (2019) 936.
https://doi.org/10.1080/00914037.2018.1525538
[3] I. Smirnova, P. Gurikov, Aerogel production: Current status, research directions, and future
opportunities, J. Supercrit. Fluids. 134 (2018) 228.
https://doi.org/10.1016/j.supflu.2017.12.037
[4] P.Sacco, M. Borgogna, A.Travan, E. Marsich, S. Paoletti, F. Asaro, M. Grassi, I.Donati,
Polysaccharide-Based Networks from Homogeneous Chitosan-Tripolyphosphate Hydrogels: Synthesis
and Characterization, Biomacromolecules, 15 (2014) 3396.
https://doi.org/10.1021/bm500909n
[5] C.S.A. Bento, S. Alarico, N. Empadinhas, H.C. de Sousa, M.E.M. Braga, Sequential scCO2 drying
and sterilisation of alginate-gelatine aerogels for biomedical applications, J. Supercrit. Fluids. 184
(2022) 105570.
https://doi.org/10.1016/j.supflu.2022.105570

97
High pressure packed column for fractionation of low volatile and
viscous mixtures. Conceptual design of glycerol acetates
refinement with scCO2
Mariana Fortunatti Montoyaa, Francisco Sanchez, Pablo Hegela, Selva Peredaa,*
a
Planta Piloto de Ingeniería Química (PLAPIQUI), Chemical Engineering Department, Universidad
Nacional del Sur (UNS) - CONICET, Camino La Carrindanga Km7, 8000B Bahía Blanca, Argentina
* spereda@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT

Glycerol esterification with carboxylic acids is a highly studied chemical route to add value to
renewable byproducts in biorefineries. This is a stepwise reversible reaction, whose
intermediate products have higher value than the final fully esterified glycerol molecule
(triglyceride). Therefore, high yield fractionation processes are needed to recover mono and
di-acyl glycerides. Like glycerol, both, low molecular weight, and fatty acids are today
available as low-cost reactants in biorefineries. Interestingly, the acetylation of glycerol
comprises the condensation reaction of two by-products from different-platform biorefineries:
glycerol from oil-based and carbohydrate- and/or lignocellulosic-based biorefineries. Acyl
glycerides are typically non-volatile, thermolabile, and viscous compounds, features that
hinder their separation and purification. Additionally, depending on the molecular weight of
the carboxylic acid, intermediate products may even show azeotropic behaviour, like
monoacetin and diacetin.
Supercritical CO2 fractionation is a convenient technology for processing these mixtures, it
operates at moderate temperatures and remarkably reduces the mass transfer resistances
typically found in viscous systems. In previous works, we demonstrated the technical
feasibility of fractionating glycerol acetates using CO2 as a solvent. Moreover, we extended the

98
GCA-EOS to accurately model, by group contribution, the phase equilibria of mixtures that
can be found in the homologous family of acylglicerides.
In this work, we carry out the conceptual design of a high-pressure fractionation column for
the rectification of intermediate products of glycerol acetylation (mono- and di-acetin) based
on GCA-EOS predictions. To challenge the model accuracy, we contrasted its predictions with
the results of experimental scCO2 fractionation of a commercial mixture of glycerol acetates,
conducted at temperatures ranging from 301 K to 323 K and pressures between 100 bar and
130 bar. Given the group contribution nature of GCA-EOS, the simulation tool developed in
this work is also valuable for designing separation processes of other surfactants.
A proof of concept was conducted in a bench-scale high-pressure packed column (19 mm ID,
total height 4 m) at 313 K and 95 bar, using a solvent-to-feed ratio (S/F) of 17.5 g CO2/g feed.
Based on these findings, the height equivalent of theoretical plate (HETP) was determined from
the number of effective equilibrium stages in the equipment and the total height of the packing
material. Both, the experimental results and the simulations show that a reflux is needed, as a
countercurrent isothermal operation leads to a low recovery of the target compounds.
Therefore, two design options are considered: (i) a countercurrent column with internal reflux
using a hot finger point, and (ii) a countercurrent column design with external reflux using a
high-pressure pump. Finally, we use the GCA-EOS to design a feasible operating window to
fractionate a commercial mixture of glycerol acetates (23.3 wt% triacetin, 48.7 wt% diacetin,
and 28.0 wt% monoacetin) in two steps, using a single column of 11 stages, and fed at stage 4.
According to our simulations, the more favourable operating conditions for monoacetin
refining are 306 K and 91 bar, with a reflux ratio of 1.6 and 65 kg CO2/kg of feed, while for
diacetin the conditions are 310 K and 93 bar, with a reflux ratio of 8 and 45 kg CO2/kg feed.

Keywords: CO2, supercritical, purification, acylglycerides, glycerol

99
 Extraction of bioactive compounds from Scaptotrigona depelis
pollen using supercritical carbon dioxide.
Adriane Gomes da Silvaa,*, Eduardo Gama Ortiz Menezesb, Raul Nunes de Carvalho Juniora
a
Federal University of Pará 1: Adriane Gomes da Silva, R. Augusto Corrêa, 01 - Guamá, 66075-110,
Belém – PA, Brazil
b
Federal Institute of Education, Science and Technology of Rondônia: Eduardo Gama Ortiz Menezes,
Avenida Calama 4985, 78908-010, Porto Velho, Rondônia, Brazil.
*corresponding author email address: adriane.dasilva.gomes@gmail.com.

GRAPHICAL ABSTRACT

ABSTRACT
In recent years the search for sustainable technologies has become an increasingly present
concern in various sectors of society and this is due to the fact that the use of natural resources
in an unbridled and unplanned way generating negative impacts on the environment and the
people who depend on it, as is the case of the original peoples who inhabit the Amazon region.
In this context, the processing of Amazonian plant matrices can be a viable and sustainable
alternative for the economic development of the region, provided that it is used consciously
and respecting the preservation of the environment.
The plant matrices of the Amazon, such as the pollen of stingless bees, which in its composition
has a diversity of bioactive, many with medicinal and even nutraceutical properties, are valued
worldwide. And the exploitation of bee pollen must be sustainable to ensure economic progress
and preservation of the environment, for this it is necessary to use technologies for the
processing of these matrices to be developed to minimize environmental impacts and the
preservation of these species. Therefore, supercritical technology is shown as a sustainable
alternative to conventional techniques, such as soxhlet and maceration that require large

100
amounts of toxic organic solvents to ensure the solubility of certain bioactive compounds.
Thus, the objective of this work is to obtain pollen extract and concentrate in polyphenols and
evaluate the influence of pressure and temperature on the extraction yield and identify or
estimate kinetic parameters that guarantee estimates of the characteristic extraction times.
The experimental methodology consists of two main steps: the first is the evaluation of the
effect of pressure and temperature through the global yield isotherms and later there will be the
evaluation of the influence of the extraction time. The overall yield isotherm will be
experimentally evaluated at pressure conditions 200,300 and 400 bar at 40°C isotherm. The
extraction kinetics will be evaluated by estimating the kinetic parameters by applying mass
transfer models described in the literature.
An initial, theoretical analysis prior to the experiments consisted of applying the Peng-
Robinson cubic equation of state to evaluate the influence of temperature and pressure on
solubility in the supercritical solvent in the plant matrix. These results showed that solubility
is influenced by pressure and temperature, which had high solubility values in the condition of
40ºC and pressure of 400 bar.

Keywords: Amazon, Stingless bees, Pollen, Sustainable, Supercritical.

101
 Production of homogeneous biofuel mixtures by supercritical
transesterification: experimental results, process simulation and
life cycle assessment.
Lizeth Molinaa, Victor F. Marulandab,*
a
Universidad de La Salle, Carrera 2 # 10-70, Bogotá, Colombia
b
Universidad del Valle, Calle 13 # 100-00, Cali, ColombiaUniversity of Pará 1: Adriane Gomes da
Silva, R. Augusto Corrêa, 01 - Guamá, 66075-110, Belém – PA, Brazil
*corresponding author email address: victor.marulanda@correounivalle.edu.co.

GRAPHICAL ABSTRACT

ABSTRACT
In the supercritical transesterification process biofuel is produced from a triglyceride source
and an alcohol at temperatures and pressures usually in the range 300 °C to 350 °C, pressures
above 13 MPa and high alcohol to oil molar ratios. At these conditions, alcohol and
triglycerides form a homogeneous phase and no catalyst is required, and triglyceride sources
such as unrefined vegetable oils, animal fats and waste oils can be directly trans esterified
without pretreatment steps. Yet, use of high alcohol to oil molar ratios is translated into a high
energy consumption and an unfavorable environmental performance. Alternatively, some
studies have dealt with the transesterification reaction at temperatures higher than 350 °C and
lower molar ratios (9:1- 15:1) and reported that esters and glycerol decomposition reactions
took place, but not necessarily conducing to the degradation of the properties of the resulting
biofuel.
In this work, the production of a biofuel mixture by supercritical transesterification of refined
palm oil and aqueous ethanol was carried out at temperatures higher than 400 °C and a 12:1
ethanol to oil molar ratio, to assess the effect of temperature and residence time in the formation
of a homogeneous phase, effluent appearance, glycerol decomposition and increased water

102
content. Glycerol decomposition was assessed by means of water production and mass balances
according to the expected glycerol etherification reactions described in literature. In a typical
run ethanol was first pumped through the system for a few minutes to purge trapped air. Once
purged from air, the needle valve was shut off and ethanol pump was stopped. Preheaters and
reactor were set at the desired reaction temperature while keeping the system pressure above
20 MPa (3000 Psi). Once the selected temperature in preheaters and reactor was reached,
ethanol and vegetable oil pumps were simultaneously started. Pressure was manually
controlled, and samples were taken under steady state conditions after at least 3 residence times
in the reactor had passed.
The results showed glycerol was formed at temperatures near 400°C and decomposed at higher
temperatures, whereas water was produced at temperatures higher than 400 °C, and that
prolonged times resulted in gas and soot formation, indicating decomposition reactions.
Collected effluents formed a homogeneous phase, and no phase separation was observed even
after 30 days of storage at room temperature conditions. Effluent samples showed an increment
in water content with increasing temperature. However, gas formation was observed at 460 °C
with a residence time of 11.3 min. Soot formation was observed at 480 °C. Residence times
longer than 20 minutes resulted in gas formation and lower water content at temperatures above
420 °C. Gas formation could be attributed to thermal degradation reactions resulting from a
longer than required residence time. Although water formation is not commonly addressed in
supercritical transesterification studies, additional water formed can be indirectly associated to
triglycerides conversion by means of a mass balance in which triglycerides (TO) react
completely with ethanol (EtOH) to form fatty acid ethyl esters (FAEE) and glycerol (Gly),
which further reacts with excess ethanol to produce glycerol ethers (TriG) and water, according
to the proposed reaction scheme

By adding reactions (1) and (2) the global reaction was obtained

Glycerol decomposition in-situ with water produced due to alcoholysis reactions could result
in a simpler process in which glycerol separation is no longer required and the resulting mixture
comprised by biodiesel, ethanol, water, and glycerol decomposition products could be obtained
as a final product in a real one-pot process, which could be directly added to diesel fuel. Fuel
mixtures formed by blending ethyl biodiesel with diesel, particularly if the fuel formulation is
completed with bioethanol or even water, have been extensively studied as a feasible
alternative for improving engine performance and emissions, such as decreased NOX
emissions, drastic smoke reduction and lower brake specific energy consumption. Analyzed
water content in biofuel samples along with mass balances indicated the optimal set of
operation conditions for producing a homogenous biofuel sample in which glycerol was
completely decomposed and gas and soot formation was avoided was 440 °C and a residence
time of 11.7 min.

Keywords: supercritical transesterification, glycerol decomposition, biofuel mixtures.

103
 Supercritical carbon dioxide processing of maritime pine
(Pinus pinaster): sorption study
João Leocádioa, Luís Fonsecab, Mara E.M. Bragaa, Hermínio C. Sousaa*
a
Chemical Process Engineering and Forest Products Research Centre (CIEPQPF), Department of
Chemical Engineering, University of Coimbra, 3030-790 Coimbra, Portugal
b
Centre for Functional Ecology-Science for People & the Planet (CFE), Department of Life Sciences,
University of Coimbra, 3004-456 Coimbra, Portugal
*corresponding author email address: hsousa@eq.uc.pt

GRAPHICAL ABSTRACT

ABSTRACT
Wood is a widely used material due to its unique properties and features including its
availability, renewability, versatility and natural beauty. However, its vulnerability to biotic
and abiotic agents can lead to deterioration and other problems. These limitations often require
protective measures to ensure its performance, long-term durability and safe usage.
The application of supercritical fluids in wood processing, in particular the supercritical carbon
dioxide (scCO2), has been exploited as a medium for the deposition/impregnation of modifying
agents, such as biocides as well as a drying method [1, 2]. The chemical and physical
interactions between scCO2, wood, and biocides play a crucial role in efficient impregnation
since scCO2 provides a pathway for biocides to penetrate the wood structure. Therefore,
understanding scCO2 sorption on wood is an important step for optimizing wood treatment
processes and enhancing impregnation efficiency. However, the sorption behavior of scCO2
depends on several factors such as pressure, temperature, and wood properties including

104
moisture content, density, anatomical structure as well as the presence of extractives that can
affect its interaction with the wood [1].
Some studies have focused on the permeability of wood to scCO2 intending to correlate the
mass transfer of impregnated biocides and few to determine the limits of pressurization and
depressurization [1, 3], but there is still a lack of information on the total amount of CO2 sorbed
by each species.
In the current work, the sorption of scCO2 in the sapwood of maritime pine (Pinus pinaster)
was investigated for the first time at a lab scale. The study employed a Central Composite
Design (CCD) that considered mild temperature conditions of 35 to 45°C, densities of 600 to
900 kg/m3, and treatment times ranging from 0.5 to 6 hours.
Small clear wood samples (~1.4 × 1.4 × 3.4 cm) were prepared and conditioned (13–17 %
moisture) before the experiments. The amount of sorbed carbon dioxide in the pine wood was
determined using a gravimetric procedure, by recording the weight changes of samples during
desorption at atmospheric pressure, after the fast release of CO2 in the treatment vessel [4]. The
obtained data were analyzed, and the mass of CO2 was extrapolated through linear correlation.
JMP® software was used for statistical analysis and to generate graphical representations that
allowed us to evaluate the relationship between temperature, density, treatment time, and the
scCO2 sorption in the sapwood of maritime pine.
The exposure of wood to a lower temperature of 35°C favored sorption at equal densities. With
a density increase from 600 to 900 kg/m3, higher scCO2 uptake was observed in both wood
exposed to lower and higher temperatures. However, above 35°C, longer exposure times did
not promote sorption, while at this temperature that was observed. The results showed a
sorption variation ranging from approximately 0.3% to 6%, with the highest percentage being
obtained at 35°C and 900 kg/m3 after 6 hours.
The determination of optimal parameters within the tested range of conditions for the scCO2
sorption in P. pinaster sapwood provided valuable insights for the efficient treatment and
subsequent biocide deposition/impregnation.

Keywords: Wood, Pinus pinaster, scCO2, sorption, CCD.

ACKNOWLEDGEMENTS
The authors acknowledge Fundação para a Ciência e Tecnologia for the financial support
through the PhD grant of João Leocádio (UI/BD/150895/2021).
REFERENCES
[1] Kjellow, A.W., Henriksen, O., 2009. Supercritical wood impregnation. The Journal of
Supercritical Fluids 50, 297–304. https://doi.org/10.1016/j.supflu.2009.06.013
[2] Zhang, J., Yang, L., Liu, H., 2021. Green and Efficient Processing of Wood with
Supercritical CO2: A Review. Applied Sciences 11. https://doi.org/10.3390/app11093929
[3] Fernandes, J., Kjellow, A.W., Henriksen, O., 2012. Modeling and optimization of the
supercritical wood impregnation process – Focus on pressure and temperature. The Journal of
Supercritical Fluids 66, 307–314. https://doi.org/10.1016/j.supflu.2012.03.003
[4] Aroso, I.M., Duarte, A.R.C., Pires, R.R., Mano, J.F., Reis, R.L., 2015. Cork processing
with supercritical carbon dioxide: Impregnation and sorption studies. The Journal of
Supercritical Fluids 104, 251–258. https://doi.org/https://doi.org/10.1016/j.supflu.2015.06.024

105
 Phase Boundaries and Copolymerization of Acrylic Acid + Butyl
Methacrylate + Carbon Dioxide under High-Pressure Single-
Fluid Initiation

Ramses S. Meleán Britoa,b, Joana E. Tasquec, Miriam C. Strumiaa,b, Facundo Matteaa,b, Juan
M. Giussic and Juan M. Milanesio*b,d
a
Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química
Orgánica. Av. Haya de la Torre y Av. Medina Allende, Córdoba, X5000HUA, Argentina.
b
CONICET, Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada
(IPQA – UNC – CONICET). Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
c
YPF TECNOLOGÍA S.A., Av. Del Petróleo s/n (entre 129 y 143), Berisso, Argentina
d
Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales. Departamento
de Química Industrial y Aplicada. Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
*
E-mail: juan.milanesio@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
The use of pressurized fluids in precipitation polymerization offers the advantage of
minimizing, or even eliminating, the need for a separation process, yielding almost pure
polymers by a simple depressurization of the system [1]. Supercritical carbon dioxide (CO2) is
the most used industrial pressurized fluid due to its low critical temperature (Tc = 31.1 °C) [2].
Pressurized CO2 can dissolve various vinyl monomers like acrylic acid (AAc) and methacrylic
acid esters.
For the proper design of supercritical polymerization reactors, it is crucial to consider the phase
behavior of the initial reactive mixture. In this context, one of the main challenges is to

106
experimentally determine the high-pressure phase scenario for the initial reactive system, to
design and define the process variables that ensure the initiation of the polymerization in a
single-fluid phase [3].
In this study, a variable volume high-pressure cell was employed to experimentally determine
the bubble pressures of the ternary mixture CO2 + AAc + butyl methacrylate (BMA) with
different BMA contents and temperatures, and with that information determine the phase
boundaries of the ternary system. Then, the synthesis of acrylic acid-co-butyl methacrylate
copolymers with different BMA concentrations were carried out in CO2.
BMA acts as a hydrophobic associative group in the polymer chain, potentially enhancing
inter- and intra- chain interactions and, consequently, changing the viscosity of aqueous
polymer solutions, compared to polyacrylic acid solutions. In this study, the viscosities of the
different solutions were measured using a rotational rheometer in buffered solutions at pH = 7
and in water at pH = 7 by adjusting the pH with sodium hydroxide. The incorporation of BMA
in the polymer chain was analyzed using spectroscopy techniques like FTIR and 1H-NMR.
At temperatures ranging from 60 to 100 ºC, a maximum pressure of 140 bar was necessary to
dissolve both comonomers in pressurized CO2 and initiate the polymerization in a single fluid
phase.
The polymers obtained in the reaction were white dry powders, and the results revealed that
the hydrophobic monomer was effectively incorporated into the polymer chain. The obtained
results also showed that increasing the BMA content from 4% to 8% w/w does not affect the
solubility of the macromolecules in the solutions at pH = 7. However, an increase in viscosity
was observed due to the presence of hydrophobic association effects as BMA incorporation
increased.
Keywords: Supercritical, rheology, associative polymer, ternary system, polyacrylic acid.

References

[1] Z. Bin, H. Hu, M. Chen, W. Liu, Synthesis of associating poly(acrylic acid) in supercritical
carbon dioxide and its solution properties, Colloid Polym Sci. 282 (2004) 1228–1235.
https://doi.org/10.1007/s00396-004-1062-y.
[2] L. Ferro, O. Scialdone, A. Galia, Preparation of pH sensitive poly(vinilydenefluoride) porous
membranes by grafting of acrylic acid assisted by supercritical carbon dioxide, J Supercrit
Fluids. 66 (2012) 241–250. https://doi.org/10.1016/j.supflu.2011.09.004.
[3] M. Menossi, J.M. Milanesio, S. Camy, S. Harrisson, M. Strumia, M. Destarac, Volumetric
properties of carbon dioxide + acrylic acid binary in the context of supercritical precipitation
polymerization, J Supercrit Fluids. 160 (2020) 104787.
https://doi.org/10.1016/j.supflu.2020.104787.

107
 Supercritical CO2-assisted impregnation of PLA films with
R-carvone. I: Effect of processing on thermal and crystallinity
properties

Patricia P. Miranda, Nicolás A. Gañan, Raquel E. Martini, María L. Goñi*

a
IPQA (UNC – CONICET), Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina.
b
ICTA, FCEFyN, Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina. Name
*corresponding author email address: laura.goni@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Polylactic acid (PLA) is a biodegradable and biocompatible material that has become of great
interest for biomedical, pharmaceutical, and food packaging applications. The incorporation of
active compounds enhances its attractivity, due to it can be used as a release device for varied
applications such as drug delivery or food preservation. In this context, the use of supercritical
impregnation technology for the development of this kind of material appears as a good
strategy. Supercritical CO2 (scCO2) treatment of polymers can promote changes in the polymer
structure and/or morphology, due to swelling and plasticizing effects induced by scCO2. These
changes depend on the compound-polymer system and also on the operating conditions and
can affect not only the amount of active compound that can be incorporated but also the release
kinetics and the final properties of the polymer. In this sense, in the framework of a project
concerning the development of PLA-based active materials, the supercritical impregnation of
PLA films with carvone, a naturally occurring compound with antimicrobial and insecticide
activity, was investigated. The influence of process variables on the impregnation yield and
release kinetics was previously studied and reported [1]. In this work, a systematic study of the
effect of the supercritical processing and the incorporation of R-(–)-carvone on the thermal
behavior and crystallinity properties of PLA films was assessed.

108
Commercial films of PLA were kindly provided by Converflex (ARCOR Group, Argentina).
R-(–)-carvone (98% purity) was purchased from Sigma-Aldrich (Germany), and industrial
extra-dry carbon dioxide (water content ≤ 10 ppm v/v) was from Linde (Argentina). The
supercritical processing of PLA was performed in batch mode, using a lab-scale high-pressure
system described before [1], and three variables were evaluated: scCO2 density (278–683 kg
m-3), temperature (40–60 °C), and depressurization rate (0.6–6.0 MPa min-1). All runs were
performed in duplicate, with a contact time of 2 h and a carvone/PLA mass ratio of 0 or 0.5.
Thermal behavior was assessed by differential scanning calorimetry (DSC) using a Discovery
DSC 250 equipment (TA Instruments, USA), heating film samples from -10 °C to 200 °C at
10 °C min-1. Main transition phenomena were observed, recording the glass transition (Tg),
cold crystallization (Tcc), and melting (Tm) temperatures, as well as the transition enthalpies
(∆𝐻𝑐𝑐 , ∆𝐻𝑚 ). The crystallinity degree (XC%) was also calculated [1].
Neat films were nearly amorphous (XC% = 3.5 %) and showed the typical behavior of PLA (Tg
~58°C, Tcc ~102°C, and a Tm ~149 °C).
Treated films with and without the addition of carvone showed an increase in the XC% that
varied from 23 to 32 %, depending on the process conditions. This increase in the crystalline
region can be explained by the plasticizing effect of scCO2, which promotes polymer chain
mobility, allowing their rearrangement into different crystalline structures. Furthermore, a
decrease in the cold crystallization phenomenon was also observed for all treated samples, with
values of ∆𝐻𝑐𝑐 in a range of 1–3 J/g, compared to a value of 19 J/g for neat samples. These
results are in accordance with the increase in XC%, mainly due to polymer chains that are able
to crystallize in neat PLA during the DSC heating, in treated samples are mainly reorganized
during the supercritical process and only a small fraction is rearranged.
Finally, impregnated samples showed a dependency of some transition temperatures with the
content of carvone, which varied from ~6 to ~30 wt% [1]. In particular, a decrease in Tm and
Tg was observed as the amount of carvone incorporated in the films increased, which is related
to the compatibility and interaction between carvone and PLA, and this behavior may be
described by thermodynamic models. On one hand, Flory-Huggings relation [2] showed a good
prediction of the melting point depression with the increase of carvone content, observing a
less pronounced decrease of Tm for carvone contents higher than 18 wt%. On the other hand,
Fox equation [3] was used to model the Tg variation. In this case, the decrease was only
observed for small amounts of carvone content (6–10 wt%), being Tg nearly constant for higher
values (18–30 wt%). Therefore, experimental data for Tg behavior was only described by the
Fox equation for carvone contents up to 10 wt%. Taking into account that the Fox equation
describes the behavior of miscible blends, and the observed behavior of the melting point
depression, these results may suggest that the miscibility limit of carvone in PLA was reached
at around 10–18 wt%.

Keywords: Supercritical solvent processing, thermal behavior, Tg depression, Melting point

depression, plasticization

References:
[1] P. Miranda et al., Journal of CO2 Utilization, 2022, 61:102029
[2] J. Sodeau, Advances in chemical physics, 1986
[3] T. Fox, Bulletin of the American Physical Society, 1956, 1:123

109
 POSTER
PRESENTATIONS

110
 Phase Behavior of ethanolic passion fruit (Passiflora edulis sp.)
bagasse extract in supercritical CO2.
Erick J. Santos de Araujoa,b, Arthur J. Oliveira Bragac, Papa Matar Ndiayec,d Julian
Martíneza,*
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, University of Campinas, UNICAMP, Campinas, SP, Brazil.
b
Federal Institute of Education, Science and Technology of Bahia, Barreiras, Bahia, Brazil.
c
Chemical and Biochemical Process Engineering/School of Chemistry, Federal University of Rio de
Janeiro, Rio de Janeiro, RJ, Brazil.
d
Chemical Engineering Program/COPPE, Federal University of Rio de Janeiro, Rio de Janeiro, RJ,
Brazil.
*corresponding author email address: erickjarles@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
The investigation of phase behavior can be used to improve supercritical fluid impregnation
(SFI) processes, since the efficacy of this technique is closely dependent on the solubility of
the target compounds in supercritical CO2 (sc-CO2). Knowing the solubility is crucial,
considering that the saturation of the target compounds in sc-CO2 must be achieved, thus
inducing their interaction with the solid matrix and consequent impregnation. The present work
aimed to evaluate the phase behavior of a system composed by ethanolic passion fruit bagasse
extract (PFBE) and sc-CO2, in order to find the best conditions to solubilize the PFBE in a SFI
process. The passion fruit bagasse, which consisted of seeds and residual pulp, was dehydrated,
ground, and subjected to sc-CO2 extraction at 40 MPa and 35 °C in order to be defatted. To
obtain the PFBE, the defatted passion fruit bagasse was then placed in a stainless-steel cell and

111
subjected to pressurized liquid extraction (PLE) with absolute ethanol (with purity ≥ 99.5%) as
solvent at 10 MPa and 75 °C. After a static time of 10 min, dynamic extraction was performed
and the ethanolic PFBE was collected in amber glass vials for 120 min. Phase behavior of the
system PFBE + CO2 was evaluated using the static synthetic method in an experimental unit
composed of a high-pressure cell with variable volume, two high-pressure syringe pumps, two
thermostatic baths, a magnetic stirrer, a digital microscope, a gas cylinder, an analytical balance
and a control panel with valves and connections. The PFBE was introduced in the high pressure
cell using a sampling cylinder and a high-precision balance. The cell was closed, connected to
the process line, and a predetermined mass of CO2 was injected. Four different mass
compositions of the system (PFBE + CO2) were used: 5% PFBE + 95% sc-CO2, 7,5% PFBE +
92,5% sc-CO2, 10% PFBE + 90% sc-CO2, 20% PFBE + 80% sc-CO2. The system pressure
(37.5 MPa) and temperature (65 °C) were selected after previous investigations. Phase behavior
of the system was observed using a digital optical microscopy device attached to a sapphire
window in front of the HPC. According to the observed results, only one composition (5%
PFBE + 95% sc-CO2) remained stable, showing a single-phase region. In the other
compositions a solid phase was observed before the system had reached the selected pressure
and temperature conditions. This behavior was maintained even after the system achieved the
required temperature and pressure under constant agitation for 60 minutes. The solid phase
observed in the unstable compositions is the passion fruit bagasse separated from the solvent
(ethanol). This behavior can be explained by the high affinity between CO2 and ethanol
contained in the PFBE. PFBE precipitation is considered a limiting factor for the SFI process,
as it is related to the solubility of the target compounds in sc-CO2. Therefore, among the results
obtained, the best system composition to perform SFI is 5% PFBE + 95% sc-CO2.

Keywords: supercritical impregnation; static-synthetic method; process optimization.

112
 Ultrasound-assisted pressurized liquid extraction from tucumã-
do-Amazonas almonds (Astrocaryum aculeatum) defatted with
supercritical CO2
Mariana de P. Kraüss Ferreiraa,*, Luciedry M. S. Carvalhoa, Julian Martíneza
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, University of Campinas, UNICAMP, 13083-862, Campinas, SP, Brazil.
*corresponding author email address: m241258@dac.unicamp.br.

GRAPHICAL ABSTRACT

ABSTRACT
The Amazon region has a rich biodiversity in its fauna and flora, with typical species of
scientific, economic, nutritional and medical interest. Tucumã-do-Amazonas (Astrocaryum
aculeatum) is a poorly explored Amazonian fruit, extremely rich in bioactive phenolic
compounds such as quercetin and gallic acid. Besides the fruit, there is also a lack of studies
on its by-products, especially the almond. The tucumã-do-Amazonas almond can be consumed
fresh or used as raw material for the extraction of fat, which represents about 30-36% of its
weight and is edible. However, little is known about the non-fatty compounds present in the
almond. Therefore, the recovery and characterization of these compounds can be attractive,
and ultrasound-assisted pressurized liquid extraction (UAPLE) is a promising technique for
this goal. UAPLE is considered more sustainable than the traditional extraction methods and

113
its use also has the advantages of easy maintenance and low cost, due to the relatively short
extraction time and and lower solvent consumption. In the present work UAPLE was
performed to recover bioactive compounds, such as phenolics with antioxidant potential, from
defatted tucumã-do-Amazonas almonds. The undefatted almond was characterized and
presented 37,30 ± 0,38% of moisture and 34,83 ± 0,34% of lipids on a dry basis (D.B.). The
almonds were dried and the fat was extracted by supercritical carbon dioxide extraction (SFE)
at 250 bar and 40 ºC, since this technique is also concerned with green chemistry. The defatted
almonds were then subjected to UAPLE under different conditions and the extracts were
characterized in terms of global yield, total phenolic content and antioxidant capacity by FRAP
and ORAC. In the UAPLE extraction, the effects of temperature (70, 90 and 110 ºC) and
ultrasound power (160, 480 and 792 W) were investigated with a complete factorial design,
totaling 9 different conditions, keeping constant the solvent composition (ethanol 75% in
water), pressure of 100 bar, extraction time of 20 minutes and solvent flow rate of 5 mL/min.
The best UAPLE temperature and power conditions, disregarding the energy expenditure, were
110 ºC and 792 W, as expected, since these were the highest temperature and power among
those evaluated. However, some peculiarities for each analysis were observed. At some
conditions in which lower energy was required, results were statistically equal to those obtained
at 110 ºC and 792 W, such as: i) For global yield, 70 ºC and 160 W; ii) For the total phenolic
content, 90 ºC and 480 W; iii) For the antioxidant capacity by FRAP, 90 ºC and 792 W and;
iv) For the antioxidant capacity by ORAC, 70 °C and 480 W. It was expected that global yield
would increase with temperature and ultrasound power. However, these variables did not affect
global yield significantly. A possible explanation is that the used S/F (mass ratio between
solvent and defatted almond) was high enough to deplet the almonds at other conditions.
Furthermore, in longer extraction times or under high temperature and ultrasound power, free
radicals may have been be generated due to the use of ultrasound, thus interfering in the
extraction efficiency. The use of UAPLE to extract bioactive compounds proved to be efficient
mainly due to the amount of recovered compounds and the shorter time required for their
extraction, when compared to traditional techniques. At the best UAPLE condition, the content
of total extracted phenolic compounds was 6,31 ± 0,93 mg GAE (gallic acid equivalent)/g DA
(defatted almond) B.S. (dry basis); the antioxidant capacity determined by FRAP was 25,71 ±
0,36 mg TE (Trolox equivalent)/g DA B.S and by ORAC it was 42,07 ± 1,96 mg TE/g DA
B.S. It can be concluded that this work found favorable UAPLE conditions to recover bioactive
compounds from a food by-product that was defatted by SFE, valorizing a residue of an
Amazonian fruit that had not been deeply investigated before. This work characterized and
quantified the phenolic compounds and antioxidant capacity obtained by this emergent
extraction method, finding the best extraction conditions, thus encouraging the use of UAPLE,
as it is more sustainable than traditional extraction techniques.

Keywords: amazon fruit, Tucumã-do-Amazonas, sustainability, ultrasound, high pressure.

114
 Supercritical CO2 fractionation of passion fruit by-products
extract using zeolite 13-X to concentrate phytosterols and
squalene
Luana C. dos Santos1,*, Renata B. Silva2, Eupídio Scopel3, Tahmasb Hatami4, Camila
Rezende3, Julian Martínez2
1
BioEcoUva Research Institute on Bioeconomy, High Pressure Process Group, Department of
Chemical Engineering and Environmental technology, Universidad de Valladolid, Valladolid 47011,
Spain
2
Laboratory of High Pressure in Food Engeering, Departament of Food Engineering and Technology,
Faculty of Food Engineering University of Campinas, UNICAMP, Campinas – SP, Brazil
3
Institute of Chemistry, Physical Chemistry Department, University of Campinas, Unicamp, 13083-
970, Campinas, SP, Brazil
4
Department of Materials Engineering and Bioprocess, School of Chemical Engineering, University
of Campinas – UNICAMP, Av. Albert Einstein 500, CEP 13083-852 Campinas, SP, Brazil
*corresponding author email address: julian@unicamp.br

GRAPHICAL ABSTRACT

ABSTRACT
Phytosterols and squalene are two plant-derived chemicals that have gained significant interest
in the pharmaceutical and food industries, with analgesic, immune regulation, anti-
inflammatory, antioxidant, anticancer, antiaging, and even tumor- and cardio-protective
properties (Zhang et al., 2022; Spanova et al., 2011). In addition, squalene is an important
adjuvant in vaccines (Mendes et al. 2022). Squalene and phytosterols can be fractionated from
waste produced by the vegetable oil or fruit and vegetable processing. One of the best sources
for this purpose in Brazil is passion fruit by-products (PFBP), since the country produces
700,000 tons of yellow passion fruit annually (dos Santos et al., 2021). Squalene and
phytosterols can be fractionated from PFBP using either an organic solvent or a supercritical
fluid, with the supercritical fluid offering advantages such as the use of green solvents (CO2),

115
a mild temperature condition, high efficiency, high selectivity, and easy separation of the
extract from the solvent (Reverchon and De Marco, 2006). Although SFE is a promising
technology for PFBP extraction, it must be combined with a suitable post-processing procedure
to concentrate the target components. In a related work, Hatami et al. (2020) investigated the
combination of supercritical extraction and supercritical adsorption techniques from PFBP to
produce an extract containing a specific class of chemicals. The present work modifies Hatami
et al. (2020) previous work, in a way to concentrate phytosterols and squalene in the extract.
For this purpose, PFBP was fed into a supercritical fluid extraction (SFE) column at 40 ºC and
35 MPa to obtain an extract. The extract was then placed in a stainless steel solubilization cell
connected to a supercritical adsorption column packed with zeolites. The extract solubilization
was fixed at a CO2 flow rate of 1.06 × 10-4 kg/s, pressure of 35 ± 2 MPa and temperature of 40
± 2 ºC. After extract solubilization, the mixture was carried to a fractionation column filled
with a compacted bed of zeolites 13-X with 5, 10, or 15 g. The column was submerged in a
water bath at 50, 60, or 70 ºC. Each mass × temperature binomial was performed twice. The
static time in the solubilization vessel was 45 minutes. The collected fractions were accounted
for an extract yield kinetics (0-180 min), where the extract collected in the first 40 min was
determined as F1 and the extract lasting to 180 min, F2. The mass of zeolites of 15 g in the
adsorption column was the best to concentrate all phytosterols (in F2) and squalene (in F1) at
50 ºC, achieving concentrations of 1346.55 and 1630.56 mg/kg extract, respectively. In
comparison to the raw extract composition, the adsorption fractionation under such condition
was able to successfully concentrate squalene, β-sitosterol and stigmasterol in two times for
each compound.

Keywords: passion fruit by-products, supercritical CO2, adsorption, squalene, phytosterols.

References :
R.Y. Zhang et al., J Agr Food Chem, 2022, 70, 2483-2494
M. Spanova, G. Daum, Eur J Lipid Sci Technol., 2011, 113, 1299-1320
A. Mendes et al. Pharmaceuticals, 2022, 15, 265
L.C. dos Santos et al., J. Supercrit. Fluid., 2020, 168, 105093
E. Reverchon, I. De Marco, J. Supercrit. Fluid., 2006, 38, 146-166
T. Hatami et al., 2020, J. Supercrit. Fluid., 2020, 162, 104856

116
 Production of vitamin C-vitamin B3 cocrystals by the gaseous
antisolvent (GAS) technique
Clóvis A. Balbinot Filho *, Jônatas L. Dias, Evertan A. Rebelatto, Sandra R. S. Ferreira,
Marcelo Lanza

Universidade Federal de Santa Catarina, Department of Chemical and Food Engineering, PO Box
476, 88040-900, Florianópolis-SC, Brazil
*corresponding author: clovis.filho@posgrad.ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Vitamins are essential nutrients with specific physiological functions. L-ascorbic acid (Vitamin
C, VitC) and nicotinamide (Vitamin B3, Nic) are crucial in maintaining health. VitC prevents
scurvy, promotes tissue repair, and possesses strong antioxidant properties associated with
cancer prevention. Nic is an anti-pellagra vitamin and is effective in treating neurodegenerative
diseases. The development of multivitamin cocrystals, organized crystal structures formed by
molecules with complementary hydrogen bonding sites, offers a new approach for delivering
drug and nutrient compounds in pharmaceutical and food applications. In this context,
cocrystallization employing pressurized carbon dioxide (CO2) as an antisolvent stands out for
unique morphology and physicochemical properties of the the resulting powders. This study
explored the production of VitC-Nic cocrystals by the gaseous antisolvent (GAS) process using
ethanol as solvent. Initially, GAS process was conducted at 45 °C, 90 bar, and CO2 flow of 10
mL/min. During GAS process, small and voluminous cocrystal particles are precipitated inside
the chamber as the CO2 drags out the ethanol that solubilizes the solutes. The same cocrystals

117
were obtained through conventional liquid-assisted grinding (LAG) by maceration for 10 min
of a 1:1 molar rate, adding a small volume of ethanol (200 uL). The obtained powders were
characterized by diffractive powder X-ray (DRX) and differential scanning calorimetry (DSC),
and the resulting spectra were compared with pure compounds and physical mixture to confirm
cocrystal formation. A Box-Behnken experimental design (15 runs) was employed to evaluate
the following GAS process variables: temperature (35, 45, and 55 °C), CO2 flow rate (5, 10,
and 15 mL/min), and the molar ratio of VitC:Nic (1:2; 1:1 and 2:1). The mass yield, the
antioxidant activity (AA), determined by the DPPH method and the particle size of the obtained
powder measured through Scanning Electronic Microscopy (SEM) images of each run were
used as the responses. DRX diffraction peaks of the cocrystals and parent compounds showed
peaks of higher intensity at angles (2θ) that coincided with pure standards in the literature. DSC
thermograms of pure VitC and Nic showed endothermic events at 195 and 130 °C, respectively,
regarding their melting temperatures (Tm). The VitC-Nic cocrystal had an intermediary Tm of
approximately 145 °C, which differs from the mixture's eutectic point, thus indicating that a
new structure has been formed. GAS cocrystals presented a varying yield (30,35-73,85 %)
which was only negatively affected (p<0.05) by temperature. The AA varied from 1.76-4.83
mmol Trolox Equivalents (TE), which means that cocrystals could retain the AA of pure
Vitamin C (7.85 mmol TE), highly affected by the molar ratio between VitC and Nic, i.e., the
VitC content. From SEM images (2,500x magnification), particle length ranged from 4.05-
12.78 μm, presenting mainly plate and needle morphologies. DRX spectra indicated that two
distinct polymorphs were obtained (Form I, when using 1:1 or 1:2 mixtures, and Form II,
when using a 2:1 proportion), independently of the other variables. Further, the experimental
design was optimized to render higher yield and AA with smaller particle sizes using the
desirability function of Statistic 12 software. At the optimum point (40 °C, 7.5 ml/min, and 2:1
molar ratio), VitC-Nic cocrystals showed a 60.25±2.16 % yield and 4.92± 0.56 mmol TE,
which is close to the predicted values by the desirability function (67.98% and 5.01 mmol TE),
and validates the GAS experiments. The scalability of the process was verified by doubling the
feed solution and increasing the scale from 1 to 6 mmol of each starting compound, resulting
in yield increments up to 86,24%. GAS is an attractive, reproducible, and scalable method to
produce solvent-free vitamin cocrystals with improved properties that might exert beneficial
health effects when consumed. Compared to conventional (LAG) techniques with few
controllable variables, GAS-produced cocrystals can modulate their properties to achieve a
specific purpose by changing operational processes. Nonetheless, the product obtained in this
study can serve as a model role for other vitamin-vitamin food-grade cocrystals.

Keywords: Cocrystallization, vitamin-vitamin cocrystals, supercritical carbon dioxide,

antioxidant activity, yield.

118
Effect of temperature and solvent composition on the pressurized
liquid extraction of phenolic compounds from spent hop
Amanda M. B. Oliveiraa*, Juliane Viganób, Julian Martíneza
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and Technology,
Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas, SP, Brazil
b
Center for Natural Sciences, Federal University of São Carlos, highway Lauri Simões of Barros, km
12 – SP 189, Buri, SP 18290-000, Brazil

*corresponding author email address: amandamellissa92@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
The brewing industry produces billions of liters of beer annually and therefore contributes to
the generation of large amounts of waste such as grains, hops, and yeast. During the process,
only 15% of the hop components are effectively retained in the beer, generating a large amount
of waste known as spent hops. Spent hops are rich in bioactive compounds such as essential
oils, polyphenols, and bitter acids, which have several biological activities, including the
potential for the prevention and treatment of diseases, as well as interesting sedative properties
for application in the treatment of insomnia and sleep disorders. The demand for greener
processes based on environmental and safety aspects has encouraged the development of new
alternative techniques for extracting phytochemicals. Among these technologies, Pressurized
Liquid Extraction (PLE) stands out. PLE is a technique widely used in the recovery of polar
bioactive compounds, which can achieve high yields in short extraction times and lower solvent
and energy expenditure when compared to conventional techniques. The objective of this work
was to obtain extracts rich in phenolic compounds from the spent hop generated in the dynamic

119
dry-hopping stage of beer production. PLE was performed at 10 MPa for 20 min. Temperatures
of 70, 90, and 110 °C, and solvent compositions of 50, 75, and 100 (wt.%) ethanol in water
were evaluated. The extraction cell was loaded with 5 g of sample, heated, and the solvent was
pumped until the cell was filled. After 15 min of heating to operation temperature and 10 min
of static time, the dynamic extraction was started. After extraction, the mixture of solvent and
extract was decompressed in a micrometric valve. Extracts were stored in amber bottles at -18
°C until analysis. The extracts were analyzed in terms of global yield, total phenolic content
(Folin-Ciocalteu), and antioxidant capacity (FRAP and ORAC). The PLE global yield varied
between 7.76 and 12.06% for the different conditions evaluated. The increase in temperature
from 70 to 110 °C positively affected the global yield in the extractions with 50% ethanol,
resulting in an increase of approximately 15% (from 10.48 to 12.06%). The use of high
temperatures in PLE helps to reduce the viscosity and surface tension of the solvent, increasing
its penetration into the solid material. In addition, higher temperatures increase the solubility
of the target compounds, as well as facilitate the breaking of chemical bonds in the solid
material, releasing the solute for contact with the solvent, intensifying the mass transfer, and
therefore increasing the global yield. No significant differences (p > 0.05) in global yield were
observed in the other solvent compositions. Regarding the solvent composition, significant
differences (p < 0.05) were only observed at 110 °C, with a reduction of approximately 22%
in the global yield (from 12.06 to 9.41%) between the conditions of 50 and 100% ethanol. The
selectivity of the solvent deeply depends on its physical-chemical properties and its interaction
with the solutes of interest. The chosen solvent should improve the release and solubilization
of the target compounds, even if they are in low concentrations. The total phenolic contents
obtained in PLE from spent hops ranged from 4.15 to 6.90 mg GAE (Gallic Acid Equivalent)/g
of spent hops. No significant differences (p > 0.05) were observed for the different
temperatures, suggesting that the lowest temperature applied (70 °C) was sufficient for the
recovery of an extract rich in phenolics. Regarding the solvent composition, no significant
differences (p > 0.05) were observed at 70 °C. However, at 90 °C there was a reduction of
approximately 20% of the phenolic content compared to those obtained with 75 and 100%
ethanol (from 6.32 to 5.06 mg GAE/g spent hops). At 110 °C, in turn, a reduction of
approximately 26% of the phenolic content was observed from 75 to 100% ethanol (from 6.90
to 5.08 mg GAE/g spent hops). Regardless of the temperature, PLE with 75% ethanol achieved
the greatest phenolic yields, proving that changes in the solvent’s polarity and viscosity
positively affect the phenolic extraction. The antioxidant capacities determined by the FRAP
and ORAC methods followed the same trend as global yield. In FRAP, no significant
differences (p > 0.05) were observed with 75% ethanol, regardless the temperature. However,
with 50% ethanol, there was an increase of approximately 50% in antioxidant capacity between
70 and 110 °C (from 5.91 to 9.02 mg Trolox Equivalent (TE)/g spent hops). With 100%
ethanol, there was a 50% increase in antioxidant capacity between 70 and 110 °C (from 4.40
to 6.60 mg TE/g spent hops). For ORAC, no significant differences (p > 0.05) were observed
between temperatures or solvent compositions. The highest phenolic yields and antioxidant
capacities were obtained at 110 °C, indicating that this temperature can be used in the recovery
of extracts rich in phenolic compounds, without a negative impact on their antioxidant capacity.
Considering the obtained yield and operation cost, PLE at 70 °C using 75% ethanol was chosen
as the best process condition, confirming that PLE is a sustainable and efficient technique that
can be used to recover phenolic compounds with high antioxidant capacity from plants and
food processing wastes.

Keywords: Agro-industrial waste, antioxidants, hops, pressurized liquids.

120
 Supercritical impregnation of phenolic compounds of passion
fruit (Passiflora edulis sp.) bagasse extract in corn starch
aerogels: effect of operation mode
Erick Jarles Santos de Araujo* a, b, Julian Martínez a
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, Faculty of Food Engineering, University of Campinas, Campinas-SP, Brazil.
b
Federal Institute of Education, Science and Technology of Bahia, Barreiras, Bahia, Brazil.
*Corresponding author email address: erickjarles@gmail.com

ABSTRACT
Supercritical fluid impregnation (SFI) is an emerging technique that has gained prominence
due to its many advantages and applications. It is consensual that, to obtain good results in SFI,
it is crucial to understand the effects of process parameters such as pressure, temperature,
contact time, depressurization rate, and operation mode. This work evaluated the effect of the
operation mode on the incorporation of polyphenols from passion fruit bagasse extract (PFBE)
into corn starch aerogels by SFI. The passion fruit bagasse, which consisted of seeds and
residual pulp, was dehydrated, ground, and subjected to sc-CO2 extraction at 40 MPa and 35
°C in order to be defatted. To obtain the PFBE, the defatted passion fruit bagasse was then
subjected to pressurized liquid extraction (PLE) with absolute ethanol (with purity ≥ 99.5%) as
solvent at 10 MPa and 75 °C. After a static time of 10 min, dynamic extraction was performed
and the ethanolic PFBE was collected in amber glass vials for 120 min. Corn starch aerogel
particles were produced in three steps. First, the hydrogel was prepared using the gel-emulsion
technique followed by retrogradation (4° C, 48 h). In the second step, solvent exchange, alcohol
gel was produced by prior removal of the oil phase from the hydrogel, followed by immersion
of the particles in ethanol/water mixtures (30, 60, 90, and 100%, v/v) at a particle-to-liquid
mass ratio of 1:5 and an exchange frequency of 24 h. Finally, aerogels were obtained by
supercritical drying of alcohol gel at 40 °C and 12 MPa. SFI was performed at the best pressure

121
(37.5 MPa) and temperature (65 °C) conditions found in previous investigation, time of 60 min,
and average depressurization rate of 1.3 MPa/min. Three operating modes were evaluated:
static, dynamic, and semi-dynamic. In the static mode, the vessel outlet was closed, upon
reaching the required conditions, to maintain a fixed amount of sc-CO2 inside it; in the dynamic
mode, sc-CO2 was renewed in the system at an average flow rate of 5 g/min to maintain the
pressure inside the vessel; in the semi-dynamic mode, the system was kept half the time in the
static mode and the other half in the dynamic mode. The results were evaluated in terms of
loading (%), defined as the mass ratio between aerogel before and after SFI, and antioxidant
capacity (ORAC). In terms of loading, no significant differences were found between the
operation modes. However, in terms of antioxidant capacity, the static mode provided the best
result (18. 724 mg TE/g aerogel), followed by the semi-dynamic mode (13.508 mg TE/g
aerogel). This behavior suggests that the system's opening leads to a partial loss of phenolic
compounds. Thus, the static mode is the most recommended for the SFI of polyphenols from
passion fruit bagasse extract in corn starch aerogels.

Keywords: impregnation; operation parameters; polyphenols; loading; antioxidant capacity.

122
 Pressurized liquid extraction of phenolic compounds from pequi
almond (Caryocar brasiliense Camb.)
Isabela F. Morenoa,*, Julian Martíneza
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and Technology,
Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas- SP, Brazil
*corresponding author email address: isabelafmoreno@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Pequi (Caryocar brasiliense Camb.) is a native fruit of the Brazilian savanna. This fruit is
composed of epicarp, mesocarp and endocarp, which is divided into tegument, almond, and
edible pulp. Due to the thorns that surround it, the pequi almond is usually discarded as waste,
since pequi processing is mostly artisanal. Pequi almond is a promising source of bioactive
compounds, mainly phenolics. Therefore, the aim of this work was to extract the phenolic
compounds from pequi almonds using pressurized liquid extraction (PLE). PLE was performed
using the sample previously defatted by supercritical carbon dioxide extraction (SFE) at 35
MPa and 40 °C. The PLE extraction was carried out with a static period of 10 min and a
dynamic period of 60 min, at a constant pressure of 10.0 ± 0.5 MPa. The temperatures used in
PLE were 60, 85 and 110 °C and the solvent consisted of mixtures of ethanol (purity ≥ 99.5%)
in water at 50, 75 and 100% (v/v), with a constant mass flow rate of 5 g/min, which resulted in
volumetric flow rates of 5.5, 5.89 and 6.37 mL/min. The content of total phenolics in the
extracts was determined by the Folin-Ciocalteu method, in which the absorbance was read at a
wavelength of 760 nm in a microplate reader FLUOstar Optima (Ortenberg, Germany). The
greatest recovery of phenolic compounds was observed at the temperature of 110 °C and

123
ethanol concentrations of 50 and 75% (v/v). The kinetic curves show that in these conditions,
the extraction of the phenolics compounds took place during the entire PLE period. This was
not observed in the other conditions, in which the extraction of these compounds stopped at
about 30 min of the PLE, resulting in lower yields. The increase in temperature contributes to
the reduction of solvent viscosity, which helps the solvent penetration in the matrix, leading to
an increase in the amount of extracted compounds. In addition, mixtures of ethanol and water
as solvents provide shorter extraction times than pure ethanol due to the changes in the polarity
of ethanol at high temperatures, causing the breaking of hydrogen and dipole-dipole bonds.
Therefore, for the recovery of phenolics from pequi almond, mixtures of ethanol and water are
indicated as solvent. Furthermore, the increase in temperature allowed a better yield of phenolic
compounds. For future investigations it would be interesting to study the influence of different
PLE temperatures and solvent com positions in different discarded parts of pequi, such as the
peel.

Keywords: bioactive compounds, emerging technologies, food waste, biodiversity.

124
Recovery of bioactive compounds from sesame cake by combined
high-pressure methods
Marta A. de Avila Souza, Adenilson R. Rudke, Sandra R. Salvador Ferreira*

Chemical and Food Engineering Department, Federal University of Santa Catarina, EQA/UFSC, C.P.
476, CEP 88040-900 Florianópolis, SC, Brazil.
*corresponding author email address: s.ferreira@ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Nowadays, the risk of exhaustion of natural resources and raw materials contribute to the
increase in food waste recycling. The production of sesame oil, commonly by cold pressing,
provides the defatted sesame cake as a coproduct. Sesame seeds contain diverse lignans and
phenolic compounds with important antioxidant potential, as previous attributed to lignan
glycosides. Besides, the protein fraction from sesame cake can stabilize colloidal systems such
as foam and emulsions, with potential for use as food ingredient. However, sesame cake is
underestimated and mostly used as animal feed or discarded as waste material. Therefore, the
valorization of this cheap and easily available residue is social, economic, and ecologically
relevant. The recovery of antioxidant compounds from sesame has been mostly carried out by
low-pressure methods (maceration, Soxhlet) with solvents such as methanol, ethanol, acetone,
diethyl ether, and ethyl acetate, although these methods have some drawbacks, including long
processing times, large amounts of solvents, and high process temperature, affecting the quality
of the recovered extract. Alternatively, Supercritical fluid extraction (SFE) and Pressurized

125
liquid extraction (PLE) can provide advantages, like easy analyte recovery, high diffusivity
and low processing temperature (SFE), adequate for thermally labile or easily oxidizable
compounds. PLE improves solvation power and mass transfer, resulting in high yields with
low solvent use. Therefore, the objective of the present study was to combine SFE and PLE for
sesame cake fractionation (recovery of oily and polar fractions). The raw material (sesame
cake) was submitted to Soxhlet extraction with hexane at 1:30 (solid to solvent ratio), 60°C
and 6 hours, providing and oily fraction yield of 32.78% (g/g). The oily fraction was also
recovered by SFE with CO2 at 50°C and 220 bar for 150 min and CO2 flow rate of 1 kg/h,
reaching an SFE yield of 26.77 % (g/g). The recovered oily fractions (by Soxhlet-hexane and
by SFE-CO2) were evaluated by gas-chromatography showing elevated content of essential
fatty acids, rich in linoleic and linolenic acids. Nevertheless, SFE sample was obtained using
low temperature and processing time, compared to the Soxhlet method. Then, after recovering
the oily fraction by SFE, the resulting solid material was submitted to PLE at 100 bar, 55°C
and solvent flow rate of 4mL/min, with ethanol:water solution of 25% (w/w) as solvent, for
polar fraction recovery. The PLE conditions were based on previous study from the research
group. PLE method provided yield of 15.97%, and the recovered extract presented TPC of
22.67 mgGAE/g, DPPH of 97.30 μmolTE/g, and protein content of 83.68 mg/100g. The results
show the relevance of combining SFE and PLE, for the recovery of two fractions from sesame
cake: an oily fraction by SFE, rich in essential fatty acids, and a polar fraction by PLE, with
high free radical scavenging capacity (DPPH assay) and relevant protein content. Combining
SFE and PLE evidenced the sesame cake relevance, confirming the nutritional importance of
this coproduct and the effectiveness of the high-pressure methods employed.

Keywords: defatted sesame cake, supercritical CO2, pressurized liquid extraction, bioactive
compounds, antioxidant activityA

126
 Optimization of pressurized liquid extraction used to recover
flavonoids present in testa shell from Anacardium Occidentale L.
Jonas da Silvaa, Edy Sousa de Britob, Sandra Regina Salvador Ferreiraa,*

a
Department of Chemical and Food Engineering, Federal University of Santa Catarina,
EQA/CTC/UFSC, 88040-900, Florianópolis - SC, Brazil.
b
Embrapa Alimentos e Territórios, Maceió, AL, 57020-050, Brazil.
* Corresponding author: s.ferreira@ufsc.br (Sandra R. S. Ferreira)

GRAPHICAL ABSTRACT

ABSTRACT
Cashew industries generate, daily, significant amount of waste materials, mainly from
juice and cashew nut production. Testa shell is a by-product discarded after cashew processing,
which can cause environmental and economic problems. This by-product contains several
bioactive substances, such as phenolic compounds, including different valuable flavonoid
components. Pressurized liquid extraction (PLE) was employed to recover these relevant
components. This technique was optimized using a rotational central composite design to
evaluate the effect of the independent variables, Temperature (T ºC) and ethanol concentration
(EtOH %) in aqueous solutions, in terms of process yield and total flavonoid content (TFC)
from the recovered extracts. Each variable had five levels, respectively: -α minimum axial
value (29.29 °C and 7.57 %), -1 minimum value (50.00 °C and 20.00 %), 1 central point
(100.00 °C and 50.00 %), +1 maximum value (150.00 °C and 80.00 %), and +α maximum axial

127
value (170.71 °C and 92.43%). Combining the different variable levels and the central point
repetitions, 11 assays were conducted at 100 bar for PLE optimization. The PLE yields varied
from 20.26 to 71.93 %, and the TFC, determined as determined by Zhishen et al., varied from
197.91 to 373.61 mg CAE g-1 (Catechin equivalents per gram of dried extract). These values
indicate that testa shell is a good source of flavonoid components due to the high recovery by
PLE method. The influence of the PLE parameters on process yield and TFC values was
modeled using a second-order regression equation, with a good fit to all experimental data. The
proposed regression model shows an R² of 0.9065, according to the analysis of variance
(ANOVA). Considering a desirability function combining process yield and TFC responses,
the optimal condition described by the model was 29.29 °C and 92.42 % of ethanol
concentration on solvent mixture. Finally, the optimized conditions show that PLE was an
adequate method to obtain flavonoids from testa shell, as a promising approach for food
processing. By applying suitable pressure, temperature, and solvent conditions, it is possible to
maximize the efficiency of the extraction, obtaining extracts rich in flavonoids with potential
application in the food, cosmetic, and pharmaceutic industries.

Keywords: Phenolic compounds, GRAS solvents, Desing of experiments

128
 Supercritical CO2 extraction of essential oil from guarana
(Paullinia cupana) peel

Rayanne P. F. de Meloa,*, Rafael C. Moreirab,, Juliano L. Bicasb, Nathana C. Bastosc, André

L. Atrochd Tatiane P. De Souzac, Julian Martineza

a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas-SP,
Brazil.
b
Laboratory of Food Biotechnology, Department of Food Science and Nutrition, Faculty of Food
Engineering, University of Campinas, UNICAMP, Campinas-SP, Brazil.
c
Laboratory of Innovation and Development in Pharmaceutical Technology, Faculty of
Pharmaceutical Science, Federal University of Amazonas, UFAM, Manaus-AM
d
Guarana Breeding Laboratory, EMBRAPA Western Amazon.

*rayannepfm.94@hotmail.com

GRAPHICAL ABSTRACT

129
ABSTRACT

Brazil is the largest producer of guarana (Paullinia cupana), a native fruit of the Amazon
rainforest, usually found in the southeastern region of the state of Amazonas. The guarana
fruit is divided into pulp, seed, and peel. Guarana seeds are used in the production of
beverages and other products, so peels (accounting for c.a. 30% of the fruit’s weight) and pulp
are processing side-stream products, which may lead to adverse economic, ecological, and
social impacts. Waste generation is an economic, environmental, and social problem. Guarana
peels contain essential oils and biologically active compounds that can be applied in food,
pharmaceuticals, and cosmetics. These compounds can be extracted with Supercritical Fluid
Extraction (SFE) using CO2 (carbon dioxide) as solvent. SFE is attractive for extraction
processes, as it is considered clean because it does little harm to the environment and is inert
to the extracts at the end of the process. Therefore, this work aimed to extract essential oil
from guarana peel by SFE and identify its main compounds. Guarana dried peels were kindly
donated by Guarana Breeding Laboratory of EMBRAPA Western Amazon. The peels were
dried in an air circulation oven at 45 °C and ground in a knife mill with a mesh opening of 1
mm. The dried peels were subjected to SFE at a fixed temperature of 40 °C and pressures of
25.0, 30.0, and 35.0 MPa, with a CO2 flow rate of approximately 3.6 g/min. Each experimental
condition was performed in duplicates. The SFE global yields were evaluated for all the
conditions. Gas Chromatography Coupled with Mass Spectrometry (GC-MS) was performed
to identify the main compounds of the extract. The global yields obtained after the extractions
at 40 °C were 2.40%, 2.80, and 3.60% at 25.0, 30.0, and 35.0 MPa, respectively. Thus, the
highest working pressure (35.0 MPa) obtained the highest global yield. This was already
expected since, as pressure increases, the density of the solvent increases and, consequently,
its solvation power, leading to the extraction of a greater amount of compounds that have a
chemical affinity with the solvent. In the extracts obtained at 35.0 MPa, compounds such as
caryophyllene, humulene, and copaene were identified by GC-MS. These compounds are
sesquiterpenes, which confirm that guarana peels contain essential oil. Caryophyllene is
directly to the prevention of diabetes, cerebral ischemia, depression, and Alzheimer's disease.
Caffeine, an important stimulant for the human body, was also identified in the extract.
Therefore, we can conclude that the essential oil of guarana peel can be extracted by means
of SFE, and it contains compounds with applicability in food and pharmaceutical products.

Keywords: Essential oil, guarana, food waste, biodiversity.

130
 Fractionation of α-terpineol and limonene by supercritical fluid
adsorption
Rayanne P. F. de Meloa,, Beatriz B. Kienasta, Luana C. dos Santosb, Rafael C. Moreirac,,
Juliano L. Bicasc, Julian Martineza
a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and Technology,
Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas-SP, Brazil.
b
BioEcoUva Research Institute on Bioeconomy, High Pressure Process Group, Department of
Chemical Engineering and Environmental technology, Universidad de Valladolid, Valladolid 47011,
Spain
c
Laboratory of Food Biotechnology, Department of Food Science and Nutrition, Faculty of Food
Engineering, University of Campinas, UNICAMP, Campinas-SP, Brazil.
*rayannepfm.94@hotmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Brazil is one of the world's largest exporters of oranges, with the state of São Paulo as the
leading producer. Generally, this production is destined to fresh consumption and industrial
processing. Consequently, tons of waste are generated. In these residues, essential oils contain
an extremely important component, limonene, and α-terpineol as one of its secondary
components. Limonene is part of the cyclic monoterpene family found in citrus essential oils.
From limonene, important derivatives can be obtained: carvone, menthol, carveol, and α-
terpineol, which are responsible for imparting aromas. These compounds have aroused great
interest from both researchers and industries. This interest, mainly concerning α-terpineol, is

131
due to its applicability as an aroma and flavor enhancer in condiments, chewing gum, sweets,
beverages, and baked goods. To obtain essential oils, industries use the cold pressing technique,
which can have disadvantages since hydrolysis and oxidation can occur during the pressing of
the residues. As an alternative, a technology that can avoid these undesirable reactions is the
use of supercritical fluids as selectiver solvents, and thus promote the separation between target
compounds and undesirable compounds. Carbon dioxide (CO2) is one of the most used solvents
in this technique because it does little harm to the environment and has good physical
properties, separating undesirable compounds and obtaining high-quality products. In this
context, this work aimed to recover enriched fractions of α-terpineol from orange essential oil
by adsorption with supercritical CO2. The essential oil + α-terpineol mixture (60.0, 40.0, m:m)
was first solubilized in supercritical CO2 (10.0 MPa and 40 °C). This mixture was produced to
simulate a possible product of the biosynthesis of α-terpineol from limonene. A kinetics study
was previously conducted to verify the exact time the fractions reached their depletion and
could be collected. In the adsorption, the effects of temperature (40, 50, and 60 ºC) and masses
of the zeolite adsorbent (6.0, 9.0, and 12.0 g) were evaluated. Analysis by gas chromatography
with a flame ionization detector (GC-FID) was performed to determine the concentrations of
limonene and α-terpineol in the recovered fractions. From the fractionation kinetics, the percent
concentration of limonene recovered at the beginning of the process ranged from 52.68 to
58.85%, while the variation of α-terpineol was from 30.54 to 35.15%. Comparing the beginning
to the end of the process, limonene concentration decreased in all conditions, while the amount
of α-terpineol increased. This may indicate that less α-terpineol was retained in the adsorption
column. Therefore, the adsorption process seems to concentrate α-terpineol, since this
compound passes through the column with the zeolite adsorbent with all the evaluated masses.
Thus, the fractionation of α-terpineol and limonene by adsorption in supercritical CO2 from a
mixture of orange essential oil + α-terpineol was achieved.
Keywords: essential oil, supercritical fluid adsorption, limonene, α-terpineol, residues.

132
 Supercritical fluid extraction of alkaloids from Cestrum plants

Fernando D. Sanchesa, Julian Martíneza,*

a
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and Technology,
Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas/SP, Brazil.
*corresponding author email address: f167506@dac.unicamp.br

GRAPHICAL ABSTRACT

ABSTRACT

Ethnopharmacology is the science that studies the link between cultures and substance use.
The associated traditional knowledge of native peoples or traditional communities about
genetic resources may show ways that will direct research about substances with medicinal
and therapeutic potential. Several genera of the botanical family Solanaceae produce
secondary metabolites with bioactive potential, such as terpenes, flavonoids, alkaloids,
steroids and saponins. Notably, tropane alkaloids have hallucinogenic action and are related
to the toxicity of some species. Nevertheless, they produce analgesic, anti-inflammatory and
antimicrobial effects. The genus Cestrum is the second largest group of the Solanaceae and
exclusively American. During times of drought, it is associated to cattle poisoning and causes
severe liver toxicity. However, it has therapeutic value in folk medicine and includes species

133
native to Brazil, such as Cestrum axillare Vell. Supercritical fluid extraction (SFE) is widely
used to remove secondary metabolites from plant matrices and is an interesting alternative to
traditional solid-liquid extraction because it is clean, versatile and more efficient for
obtaining thermolabile bioactive compounds. Aerial parts from Cestrum plants were collected
at a local forest called Mata de Santa Genebra, a federal conservation unit located in
Campinas, Brazil. The material was washed with water, selected and dried in a forced air
circulation oven in two sequential steps: 60 ºC/25 min, to ensure denaturation of proteins and
stabilization of the plant material; 40 ºC/24 h, followed by grinding with a knife mill with a
20 or 30 mesh sieve. The ground material was placed in glass jars and kept at −18 ºC until the
extraction experiments. SFE of aerial parts from Cestrum plants was performed with a
constant solvent-to-feed ratio (S/F) of 125 kg CO2/kg dry sample, 6 grams of dry sample and
CO2 mass flow rate of 8.4 g/min. At 40 ºC, pressures of 15 and 35 MPa were tested, as well
as three conditions: pure CO2 and mixtures CO2-7% (m/m) ethanol and CO2-10% (m/m)
ethanol. Experiments were run in two rounds: in the first, the dried sample was passed
through a 30 mesh sieve and ethanol at 7% (m/m) was tested as co-solvent. In the second
round, the plant material was passed through a 20 mesh sieve and ethanol was set at 10%
(m/m). The extracts were characterized for their global extraction yield, which is the mass
ratio between extract and sample subjected to extraction, and alkaloid content, which was
determined by ultraperformance liquid chromatography coupled to mass spectrometry
(UPLC-MS). The results were evaluated by statistical analysis in order to determine the
pressure and ethanol concentration condition that obtained the greatest global extraction
yield. In general, the experiments with the 30 mesh sieve resulted in greater global yields for
all the evaluated conditions. For pure CO2, higher pressure leaded to higher global yield.
When evaluating the effect of the co-solvent, the results suggest that it increases CO2’s
solvating power and therefore the extraction yield. The best global yield was achieved at 15
MPa with the mixture CO2 + 7% (m/m) ethanol: 2.38 ± 0.15%.

Keywords : Cestrum, supercritical extraction, alkaloids, ethnopharmacology.

134
Effects of temperature and molar ratio between acetic anhydride
and α-terpineol on terpinyl acetate production in SC-CO2.
Rafael C. Moreira a*, Rayanne P. F. de Melo b, Julian Martinez b, Juliano L. Bicas a
a
Laboratory of Food Biotechnology, Department of Food Science and Nutrition, Faculty of Food
Engineering, University of Campinas, UNICAMP, Campinas, SP, Brazil.
b
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, Faculty of Food Engineering, University of Campinas, UNICAMP, Campinas, SP,
Brazil.
*rafael.chelala@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
α-terpinyl acetate is used as an aroma compound in food, cosmetics, and chemical
products due to its floral-fruity scents reminiscent of lavender. In addition to its odor-active
activity, α-terpinyl acetate has antimicrobial, antioxidant, analgesic, and therapeutic properties
in neurodegenerative diseases such as Alzheimer's. Currently, this esterification reaction is
done by chemical synthetic routes. However, these methods have certain disadvantages, such
as producing undesired compounds, low quality, low efficiency, need for strong acids as
catalysts, and high temperatures, leading to an interest in biotechnological approaches for
natural production. In contrast, enzymatic synthesis of esters offers the opportunity of yielding
natural products in mild and environmentally friendly conditions when the substrates are
equally labeled as natural. Using supercritical fluids as reaction media offers advantages such
as tunable properties and improved mass transfer, enhancing enzyme-catalyzed processes.
Additionally, temperature and pressure variations can contribute to product separation,
providing flexibility in their application.

135
 The objective of this work was to understand the effects of temperature and molar ratio
on the esterification of R-(+)-α-terpineol with acetic anhydride mediated by Lipozyme 435
lipase for the production of α-terpinyl acetate in a supercritical medium (SC-CO2).
For the reactions, R-(+)-α-terpineol (Sigma-Aldrich 96%) and acetic anhydride P.A.
(Ecibra 97%) were used. The reaction unit used in all tests consists of a CO2 pump (Maximator
M-111, Zorge/Germany), a solvent reservoir, an ultrathermostatized cooling bath (Marconi
S.A., Campinas-SP/Brazil), a thermostatic heating bath (Marconi S.A., Campinas-SP/Brazil),
pressure gauges (Zurique, São Paulo-SP/Brazil), magnetic stirrer (IKA, RCT Basic,
Staufen/Germany), thermocouples, control valves (Autoclave Engineers, Erie/PA, USA), a
micrometer valve (Autoclave Engineers, Erie/PA, USA), and a 50 mL stainless steel container.
The experiments were set up using a central composite design with two factors
(temperature - ranging from 40 to 60 ºC - and the molar ratio between anhydride and α-terpineol
- ranging from 1:3 to 6:1), with three repetitions at the central point, totaling seven experimental
assays. The effects of the variables were calculated using the Protimiza Experiment Design
Software (http://experimental-design.protimiza.com.br). The results were analyzed using the
same software at a confidence level of 95% (p < 0.05). The experiments were randomized to
maximize the variability in responses caused by external factors.
For each test, 200 mg of the enzyme was placed inside the reactor and heated to the
respective process temperature for 30 minutes to achieve thermal stabilization and remove
residual surface water from the enzyme and reactor wall. The substrates were introduced and
pressurized with SC-CO2 at 10 MPa. The agitation was set at 600 rpm, and the volume of the
reagents was fixed at 5 mL. Each reaction was carried out for 90 minutes, and at the end of the
process, the system was depressurized at a rate of 1 MPa.min-1, and the product was collected
for analysis on a gas chromatograph with flame ionization detection (GC-FID).
For the GC-FID analysis, one microliter of the sample diluted in HPLC-grade ethanol
(1:10) containing 1% heptanol was injected into the HP-7890A instrument (Agilent
Technologies) equipped with an HP-5 column (30 m x 0.25 mm x 0.25 μm). A split ratio of
1:10 was used. The operating conditions were as follows: He as the carrier gas; injector
temperature at 250°C, detector temperature at 250°C, and the column temperature programmed
to increase from 80 to 200°C at a rate of 20°C/min with an initial waiting time of 3 minutes
and 5 minutes at the final temperature.
The peak area corresponding to α-terpinyl acetate ranged from 14.8 pA.s to 59.9 pA.s.
The triplicate at the midpoint (50ºC and a molar ratio of acetic anhydride to α-terpineol of
3.5:2) had a coefficient of variation of 9.78%, which can be explained by the low automation
of the reaction unit used, as well as minor deviations that may have occurred during enzyme
weighing and reagent pipetting steps.
The statistical analyses demonstrated that temperature significantly positively affected
α-terpinyl acetate production (p = 0.0143). Moreover, the variation in molar concentration
between the reagents did not show a significant effect (p = 0.8067). There was no statistically
significant correlation between the two variables studied (p = 0.2914). Therefore, for future
optimization tests of the esterification reaction between α-terpineol and acetic anhydride,
higher temperatures should be used to increase α-terpinyl acetate productivity. A kinetic study
of the reaction in a supercritical medium will also be carried out. As for the molar concentration
of the reagents, working with excess acetic anhydride can reduce production costs, and, in a
supercritical medium, it does not cause enzymatic denaturation due to the acidity excess in the
medium.

Keywords: esterification reactions, aroma compounds, process optimization.

136
 Recovery of phenolic compounds of the Melão-de-São-Caetano
(M. charantia) peel by high-pressure extraction methods
Marcela C. de Sousa*, Adenilson R. Rudke, Sandra R. S. Ferreira

Chemical and Food Engineering Department from Federal University of Santa Catarina
(EQA/UFSC), Florianópolis, SC, Brazil
marcela.mcs@gmail.com.

GRAPHICAL ABSTRACT

ABSTRACT
M. charantia is a species of climbing plant widely found in the Northeast region of Brazil.
Popularly known as Melão-de-São-Caetano, its fruits are rarely used, being considered an
unconventional food plant in Brazil. Although known in the world for its pharmaceutical and
medicinal properties, scientific studies from the Brazilian variety are still scarce, especially
concerning the properties and characteristics from the peel of the fruit. This work aims to obtain
bioactive extracts recovered from Melão-de-São-Caetano peel through high-pressure
extractions, compared with low-pressure methods, and using the concept of green chemistry.
Bioactive compounds were recovered using the methods Soxhlet, subcritical water extraction
(SWE) and pressurized liquid extraction (PLE) under various conditions of temperature,
pressure and type of solvent. The PLE experimental conditions were 10 MPa, temperatures of
40, 60 and 80 °C, in continuous mode with ethanol and the mixture ethanol:water 50% (also
used for Soxhlet method), flow rate 3 mL.min-1 for 50 min process. The SWE assays were
performed at 10 MPa, and at 130, 150 and 170 °C, in continuous mode with a water flow rate
4 mL.min-1 for 40 min. The extraction methods were compared in terms of process yield, while
the extracts were evaluated with respect to the content of total phenolic content (TPC), by the
Folin-Ciocalteu method, and in vitro antioxidant potential by the DPPH and β-carotene

137
methods. From the results it was verified the high yields obtained from the aqueous and
hydroethanolic extracts, up to 60%, considering the different extraction methods and
temperatures studied. The effects of SWE temperature conditions were observed in terms of
the highest phenolic values and antioxidant activity, especially the sample recovered at 170°
C. The phenolic content was 29.10 mgGAEg-1 for SWE extract at 170 °C, differing statistically
(p > 0.05) from all other treatments. The antioxidant potential evaluated by DPPH presented a
maximum of 570.46 μmol TE.g-1 and by the β-carotene method of 74.41%, also employing
SWE at 170 ºC, a value with significant difference in relation to the other extracts obtained.
PLE extracts exhibited antioxidant capacity similar to Soxhlet extracts. In general, compared
to Soxhlet method, the SWE technique showed better results, even with shorter extraction time,
and less solvent volume, employing an ecologically correct method. The combination of water
and high temperature provided extracts from Melão-de-São-Caetano peel richer in phenolic
compounds and antioxidant potential, adding value to it.
Keywords: Momordica charantia; subcritical water; pressurized liquid.

138
 CHLORELLA vulgaris FRACTIONATION BY GREEN
SEQUENTIAL EXTRACTIONS: THE POTENTIAL OF
(BIO)SURFACTANT MODIFIERS
Gina G. Gonzáleza, Sandra R. Salvadora, Cristiano J. De Andradea*
a
Chemical and Food Engineering Department, Federal University of Santa Catarina,
EQA/CTC/UFSC, 88040-900, Florianópolis - SC, Brazil.
*corresponding author email address: cristiano.andrade@ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
The microalgae Chlorella vulgaris is a sustainable source of lipids, carotenoids, carbohydrates,
and proteins, since it shows high biomass productivity and easy cultivation. It can growth from
5 to 10 times faster compared to lignocellulosic plants. The C. vulgaris components can be
used by the pharmaceutical, cosmetic, biodiesel, and food industries. In this sense, traditional
microalgal lipid extraction methods such as the Soxhlet and the Bligh and Dyer (mixture of
chloroform, methanol, and water) show the drawbacks of low selectivity, high solvent/sample
rate, solvent-contamination, time and energy-consuming, among others. Nevertheless, the use
of modifiers (usually co-solvents) can be used to improve, virtually, all extraction techniques,
since they are technically and economically easy to be implemented. Unusual modifiers include
surfactants. There is little data on the extraction assisted by surfactants (all biomass) and no
data on the biosurfactant-assisted green extraction methods applied to microalgae or any other
biomass. Therefore, the aim of the work was to investigate the supercritical fluid extraction

139
(SFE) and pressurized liquid extraction (PLE) for lipid recovery from C. vulgaris associated
with (bio)surfactants as modifiers, in particular cationic (CTAB), anionic (SDS), non-ionic
(Triton x-100) and the biosurfactants mannosylerythritol lipids (MELs) at different
concentrations. Soxhlet was used as a control (lipid extraction method), the extraction was
carried out for 4 h at 80 °C with 200 ml ethanol (99.5% v/v) as the solvent. Regarding Soxhlet
extraction, the control (no surfactant) reached 14.41% of lipids. The surfactants (Soxhlet)
subtle improved the lipid extraction yield, for instance 17.30% with CTAB at 0.1 g/l; 17.66%
and 16.0% with Triton x-100 at 0.25, and 0.5 g/l, respectively; and 15.64% with SDS 0.5 g/l.
Thus, surfactant modifiers do can enhance the lipid extraction (Soxhlet). The next steps include
the investigation of green pressurized extraction systems, in particular SFE and PLE at 40 °C
and 150, 200 or 250 bar, including the effect of aforementioned (bio)surfactants. It is expected
to have a better performance when using pressurized extraction methods, achieving greater
yields, thus proposing an extraction procedure with the use of green methods, along with
modifiers, that obtains more satisfactory extract in terms of quality and quantity, in conjunction
with lower environmental impacts.

Keywords: Microalgae, assisted extractions, green technologies, lipids.

140
 INFLUENCE OF CHEMICAL COMPOSITION ON THE
PROPERTIES AND FOAMABILITY WITH SUPERCRITICAL
CO2 IN THERMOPLASTIC POLYTHIOURETHANES.
I. Graciaa, P. Belmontea*, J.M. García-Vargasa, J.F. Rodrígueza, M.J. Ramosa.

a: Department of Chemical Engineering, Faculty of Chemical Science and Technologies, University

of Castilla-La Mancha, Ciudad Real, Spain.
*Corresponding author: Pablo.Belmonte@uclm.es.

GRAPHICAL ABSTRACT

ABSTRACT
Polyurethane is one of the most versatile and employed materials in the world. However, at the
end of their life, polyurethane products are mostly incinerated or landfilled, creating
undesirable environmental effects. In order to avoid these problems, new types of
polyurethanes, based on covalent adaptable networks (CANs) that can be reprocessed, are
being developed. Being the polythiourethanes one of the most promising CANs studied by the
moment. These materials are obtained from the reaction between thiols and isocyanates, being
this reaction described as a click reaction. On the other side, the use of supercritical CO2 to
reprocess polymers into foams it is arousing a great interest. For that reason, the synthesis and
supercritical CO2 foaming of thermoplastic polythiourethanes (TPTUS) is reported in the
present work.
Three TPTUs with different types of isocyanates and polyols (PCL_TDI, PPG_TDI,
PPG_HMDI and PCL_HMDI) were synthesized, chemically (FT-IR), thermally (DSC and
TGA), and mechanically (DMA and compression) characterized. Then, these polymers were
foamed at different conditions of pressure (100 and 150 bar) and temperature (35, 50, 65, and
80 ºC). Obtaining the most relevant parameters from these foams (expansion ratio, shrinking,
cell density and average cell size). After FT-IR characterization, it was confirmed that no bands

141
in the wavenumber corresponding with isocyanate (2250 cm-1) or thiol (2540 cm-1) appear
after reaction, corroborating that all reagents have reacted completely. In addition, it was also
observed that the type of isocyanate has a large influence on the crystallinity and stiffness of
the polymers synthesized, while the type of polyol modifies the FT-IR spectra and the
decomposition curve of the material with temperature.
On the other side, once the foaming tests were performed, it was observed that the polymeric
foams with TDI as isocyanate were able to achieve higher values of expansion ratio, although
they also suffered higher shrinkage. Moreover, these foams presented cells with higher size
and heterogeneity, while the foam obtained with HMDI as isocyanate presented cells with
lower size and heterogeneity but with higher cell density. Proving that the selection of the
isocyanate type not only has a huge influence on the properties of the polymeric material but
also on the foams obtained from it.
Keywords: TPU, Polythiourethane, Foaming, Supercritical CO2

142
 Assessment of pressurized solvents extractions from grape
pomace residues
Vanessa S. Carvalhoa, Lucas C. Freitasa, Marcos L. Corazzaa,*
a
Institution: Department of Chemical Engineering, Federal University of Paraná, Av. Cel. Francisco
H. dos Santos n. 100, Curitiba-PR, Brazil
*corresponding author email address: corazza@ufpr.br

GRAPHICAL ABSTRACT

ABSTRACT
This work aims to evaluate different methods of extracting value-added products from grape
pomace (GP), a residue generated especially by the wine industry that can be reused, avoiding
environmental pollution. Recent studies revealed that GP is a potential source of bioactive
compounds like antioxidants and volatile organic compounds with beneficial effects on human
and animal health, therefore having applications in many sectors such as food, pharmaceutical
and cosmetics industries. However, it is still a challenge to find appropriate extraction
techniques that do not compromise the stability of these compounds. In this study, Soxhlet
extractions (using water, ethanol, ethyl acetate, hexane and petroleum ether as solvents), and
extractions using pressurized fluids (propane and supercritical CO2 with cosolvents) were
evaluated. The pressurized fluids extractions (PFE) were conducted in a single step, using
propane (at 100 bar and temperatures of 30, 55 and 80°C) and supercritical CO2 (at 150, 200
and 250 bar and temperatures of 40, 60 and 80°C) as solvents, and also in sequential form,
using supercritical CO2 with and without a cosolvent after the extraction made with propane
(when the cosolvent was used, the evaluated pressure range was 100 to 200 bar). The choice

143
of the cosolvent to be used was based on the best performance in terms of yield in the Soxhlet
extractions, exclusively. The experiments carried out with propane in a single step showed an
increase of 3.05 to 3.52% in the extraction yield, from the minimum to the maximum
temperature, respectively. The comparison between the extraction methods will be guided, in
addition to the yield, by the quantification of phenolic compounds, flavonoids, antioxidant
capacity (by DPPH and ABTS methods) and by the identification of volatile compounds by
GC-MS.

Keywords: Extraction, pressurized solvents, supercritical CO2, compressed propane.

144
 Supercritical fluid technology as a tool to valorize bacuri fruit
(Platonia insignis Mart.) shell
Lucas Cantão Freitasa,b, Flávia Cristina Seabra Piresa, Tiago Costa de Araújob, Raul Nunes de
Carvalho Juniora,*
a
Federal University of Pará, Rua Augusto Corrêa S/N, Guamá 66075-900, Belém-PA, Brazil
b
Federal Rural University of the Amazon, Av. Barão de Capanema S/N, Capanema-PA, Brazil
*corresponding author email address: raulncj@ufpa.br

GRAPHICAL ABSTRACT

ABSTRACT
In an era where the concept of bioeconomy and circular economy has been widely discussed
and encouraged, the application of clean technologies envisioning the use and recovery of agro-
industrial waste has been the focus of relevant studies worldwide. Currently, the agro-industrial
waste are no longer seen as a problem, but as a great ally for the development of a sustainable
industry, since they can help in the development of several products, promoting new investment
opportunities that satisfy both economic and environmental concerns, in addition to generating
income and regional development. In this scenario, the Amazon region has an enormous
spectrum of raw materials with bioeconomic potential, such as bacuri (Platonia insignis Mart.)
fruit, which is composed of up to 70% shell, a fraction often discarded as agro-industrial waste.
Therefore, the purpose of this work was to study the bacuri shell through the application of
supercritical technology in order to valorize this waste by separating its resin, which is the main
barrier for its industrial application, as well as to evaluate the extraction process parameters
studying the influence of particle size, pressure and cosolvent use. All extractions were carried

145
out at 313.15 K of temperature, 20, 30 and 40 MPa of pressure and mean particle size of 0.84,
0.50 and 0.25 mm. Sequential extractions were performed to obtain the bacuri shell extract,
where the first part of the extraction was performed only with supercritical CO2, and the second
part with the addition of ethanol as a co-solvent. The bacuri shell cake retained in the extractor
vessel was collected and analyzed by a trinocular stereoscope with image amplitude of up to
40x coupled with a high definition camera, in other to detect remaining resin in the samples.
The extracts obtained were analyzed in terms of phenolic compounds and antioxidant capacity
(ABTS). The extraction results showed that the applied technology was able to separate the
resin, which is the first report described in the literature. Furthermore, the smaller particle size
(0.25 mm) exhibited the most prominent impact on extraction rate, providing good yields of
lipid extracts (up to 10.09 ± 0.02 %) and ethanoic extracts (up to 13.78 ± 0.41%). The obtained
extracts presented good levels of phenolic compounds, which was associated with its high
antioxidant activity. Thus, the application of supercritical technology added value to the bacuri
shell, enabling new strands for industrial use of this agro-industrial waste with potential
applications in the food, pharmaceutical and cosmetic industries, encouraging the circular
economy and the bioeconomy of the Amazon region.

Keywords: Agro-industrial waste, Valorization, Bioeconomy, Resin separation, SFE

146
Pressurized propane extraction of umari (Poraqueiba sericea Tul.)
oil: extraction kinetics and fatty acid composition
Lucas Cantão Freitasa,b, Vanessa Souza Carvalhoa, Macos Lúcio Corazzaa,, Maria Lucia
Massona,*
a
Federal University of Paraná, Av. Cel. Francisco H. dos Santos n. 100, Curitiba-PR, Brazil
b
Federal Rural University of the Amazon, Av. Barão de Capanema S/N, Capanema-PA, Brazil
*corresponding author email address: masson@ufpr.br

GRAPHICAL ABSTRACT

ABSTRACT
In an era where the concept of bioeconomy has been widely discussed all over the world, the
Amazon stands out as one of the main regions with huge potential for the advent and
development of the bioeconomy. Due to its vast biodiversity and abundance, the Amazon has
a wide spectrum of raw materials with relevant bioeconomic potential, which can bring a
source of income and development to the region. In this scenario, we find umari (Poraqueiba
sericea Tul.), which is an Amazonian fruit rich in lipids with unsaturated fatty acids and beta-
carotene. However, studies reporting its scientific information are rare and, so far, no work
reports the application of innovative processes for the extraction of oil from umari fruit. Thus,
this work aimed to apply pressurized propane (PP), which is an environmentally friendly
technology with great applicability in lipophilic compounds, to extract oil from umari fruit
pulp, in addition to compare the yields obtained with conventional extraction (soxhlet) using
hexane as solvent. Before extractions, the umari pulp samples were freeze-dried and
comminuted. The parameters evaluated in the extraction with pressurized propane were flow
rate (1 to 3 mL/min) and temperature (40 to 80 °C). The pressure was kept constant at 100

147
bar. The fatty acids composition of the obtained oil was analyzed by gas chromatography
(CG-FID). Global yields, extraction kinetics and fatty acid composition from the extracts
obtained with PP were evaluated. The results obtained with pressurized propane showed that
the highest extraction yield (26.88%) was obtained at 80 °C and 3 mL/min, while the lowest
yield (23.06%) was obtained at 80 °C and 1 mL/min. In the evaluated working range, the
flow rate variation had a positive effect (p < 0.05) on the extraction yield of umari pulp, while
the temperature had a negative effect. The extraction kinetics suggest that the extraction was
predominantly carried out by solubilization of the easily accessible solute, due to the high
availability of oil on the surface of the particles. Under all conditions, more than 50% of the
oil available in the samples was extracted in the first 20 min. Moreover, the results indicate
that with 25 g of propane consumption, the yield values of all conditions were similar
(~22%), and that the mass of 40 g of propane was enough to extract most of the oil present in
the samples under all conditions evaluated. The highest yield obtained with pressurized
propane was higher than that obtained in conventional extraction by soxhlet (25.64%) using
hexane as solvent. In addition to having presented numerically higher yields, the extraction
time with propane was 70 minutes, while in soxhlet extraction it was 360 minutes, indicating
that similar yields can be obtained in a much shorter time interval by using PP. Regarding the
fatty acid composition of umari oil, oleic acid (C18:1) was predominant with 65.70% content.
Oleic acid is a monounsaturated fatty acid, classified as omega 9, which has several benefits
for human health, helping to reduce the risk of cardiovascular diseases and diabetes control.
In conclusion, extraction with PP proved to be effective for obtaining umari pulp oil,
generating high oil yields with excellent quality and great content of omega 9, which adds
value to the production chain of this species. The information on pressurized propane
parameters obtained in this work is unprecedented and should contribute to the insertion of
umari fruit in the bioeconomy scenario in the Amazon, as the oil extracted using pressurized
propane has the potential to be used in food, pharmaceutical and cosmetic industries.

Keywords: Oil, Bioeconomy, Green extraction, Kinetics, Amazon.

148
 High Pressure Liquid Impregnation of Barbatimão
Stryphnodendron adstringens (Mart.) Coville bark extracts in
biopolymeric membranes
Priscilla C. Veggia,*, Leonardo F. Camposa, Luana E. do Carmoa, Juliane Viganób,
Luciana P. Mazurc, Lucia H.I. Meic
a
Department of Chemical Engineering, Institute of Environmental, Chemical and Pharmaceutical
Sciences, Universidade Federal de São Paulo, Diadema-SP, 09913-030, Brazil
b
Centro de Ciências da Natureza, Universidade Federal de São Carlos, Rod. Lauri Simões de Barros,
km 12 - SP 189, Buri-SP, 18290-000, Brazil
c
Department of Materials Engineering and Bioprocess (DEMBio), Faculty of Chemical Engineering,
UNICAMP, Campinas-SP, 13083-852, Brazil
*corresponding author email address: pveggi@unifesp.br

GRAPHICAL ABSTRACT

ABSTRACT
Recently, worldwide concern has been raised about the environmental impact of human
activities. The use of biopolymeric materials from various sources has emerged as a promising
alternative to synthetic polymers with great potential in various industries, such as
pharmaceuticals, nutraceuticals, and cosmetics, among others. These materials are well known
for being biodegradable and originate from renewable resources. Moreover, their versatility is
enhanced as they can be molded into a variety of forms. In this regard, significant investments
are being made to broaden the utilization of natural resources and to advance scientific research
towards the incorporation of bioactive compounds. Emerging technologies, including those
that utilize high pressure, are being increasingly utilized for many applications. In particular,
the use of high pressure technology for impregnation has been extensively explored. This
technique presents an intriguing and promising alternative to traditional impregnation methods

149
for producing active polymeric materials. Nevertheless, to date, the application of High
Pressure Liquid Impregnation (HPLI) for incorporating bioactives in biopolymeric matrices
has not been addressed in the literature. HPLI is a newly developed technique for impregnating
liquid extracts into biopolymers. When the liquid extract, which is rich in bioactives, is
subjected to high pressure, it enhances the mass transfer from the solution to the polymeric
matrix in a shorter period of time. Barbatimão extract is well-known for its high antioxidant
capacity and has a wide range of pharmacological applications, such as antimicrobial and anti-
inflammatory properties. The present study aimed to advance in the field of high pressure
technologies by evaluating the efficiency of barbatimão extract impregnation in electrospun
biopolymeric membranes made of polybutylene succinate (PBS), poly(lactic acid) (PLA), and
Closite 20A. The experiments were conducted in a 100 cm3 reactor in a static mode with
varying temperatures from 45 to 60 ºC at a fixed pressure of 11 MPa for 6h. To evaluate the
process efficiency, a conventional impregnated experiment was conducted at the same
conditions in an atmospheric pressure. The impregnated membrane was then immersed in
DMSO for 24h to extract the bioactive compounds and the solution characterized in terms of
phenolic compounds (TPC). The neat biopolymer membrane and the impregnated membrane
by HPLI and conventional process were characterized by SEM (Scanning Electron
Microscope). According to results, membranes containing a phenolic compound ranging from
25.44 ± 0.13 to 43.79 ± 1.73 mg of equivalent tannic acid (ETA) per g of net membrane were
obtained. Finally, according to SEM, the pressure effect influenced in the membrane
morphology and barbatimão loading.

Keywords: Impregnation, high pressure, phenolic compound, biomaterials

150
Pressurized liquid extractions of proteins and biocompounds with
antioxidant activity from banana inflorescence (Musa acuminata
cv. Cavendish)
Gabriel Emiliano Motta, Germán Ayala Valencia, Sandra Regina Salvador Ferreira*

Chemical and Food Engineering Department, Federal University of Santa Catarina, Florianópolis, SC,
CEP 88040-900, P.O. Box 476, Brazil.
* Corresponding author. E-mail: s.ferreira@ufsc.br (Sandra R. S. Ferreira) Phone: +55 48 37212537.

GRAPHICAL ABSTRACT

ABSTRACT
Banana is one of the world's most produced and consumed fruits; however, large-scale
production results in millions of tonnes of agricultural waste left in the fields, including leaves,
pseudostems, and inflorescences. These residues can be valuable raw materials, especially
inflorescences, which have bioactive compounds of great interest to the pharmaceutical, food,
and material industries. Despite its value, these biomasses are underutilized. Therefore, the
present work aims to evaluate the extracts of the bracts of the inflorescence of Musa acuminata
Colla, from Cavendish subgroup, obtained by pressurized liquid extraction (PLE). Four mass
transfer models (diffusional, Piecewise, logistic, and Martínez) were fit to experimental
kinetics curves performed at 100°C and 10 MPa, using water, ethanol, and their 1:1 mixture as
solvents. Then, PLE optimization was performed using a full factorial design (3²), with
temperature and ethanol concentration as variables. The responses evaluated included
extraction yield, protein recovery, antioxidant capacities (DPPH, FRAP, ABTS), and total

151
phenolic content (TPC). The PLE results were compared with those from a sample recovered
by Soxhlet method and the same solvents as used at PLE. Regarding the fitting of the models:
for water, R² was greater than 0.8222 (diffusional model); for the ethanol-water mixture (1:1),
R² was greater than 0.9234 (diffusional model); and, for ethanol, R² was greater than 0.9299
(Martínez's model). In general, the PLE using water at 150 °C showed the best values of yield,
protein recovery, TPC, and antioxidant potential, compared to the aqueous extracts obtained
by Soxhlet. The PLE at optimized condition, compared Soxhlet extraction with water, showed
1.61 times higher yield, 2.06 times higher protein recovery, 3.12 times the TPC value and also
better antioxidant potential by the methods ABTS, DPPH, and FRAP. These results
innovatively indicate the use of pressurized liquids (water and ethanol) as a green technique
for recovering proteins and bioactive compounds with antioxidant activity from the bracts of
banana inflorescences.

Keywords: Residue valorization, Sustainability, Green Chemistry, Bioactive compounds,

Waste management.

152
 A model sugar cane waste biorefinery based on sub and
supercritical water processing: a LCA case study
Víctor Fernando Marulandaa,*
a
Universidad del Valle, Calle 13 # 100-00, Cali, Colombia.
*corresponding author email address: victor.marulanda@correounivalle.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
Sugar cane waste is the lignocellulosic residue comprised by leaves and tops left in the field
after mechanical harvesting and accounts up to 20% or total harvest. Valorization of these
lignocellulosic wastes, through the production of value-added products and energy, represents
a dynamization alternative for the sugar industrial sector in the medium term. A great deal of
research has been oriented towards the valorization of lignocellulosic biomasses by means of
biorefinery process schemes which incorporate acid and enzymatic hydrolysis for sugars
production followed by fermentative processes to obtain products such as ethanol and xylitol,
as well as surplus energy production from the leftover biomass fractions through combined
heat and power cycles.
More recently, subcritical and supercritical water hydrolysis has been proposed as an efficient
alternative which incorporates depolymerization, reaction and separation of sugars from
lignocellulosic biomass with high yields and short reaction times. Implementation of subcritical
water processing in a biorefinery could conduct to the formulation of simpler biorefinery
schemes with improved environmental performance than those based on conventional acid and
enzymatic hydrolysis. Therefore, in this study a sugar cane waste biorefinery based on

153
subcritical and supercritical water processes for hemicellulose and cellulose hydrolysis as well
as energy production was proposed and compared with the conventional acid and enzymatic
hydrolysis though a life cycle assessment approach.
An analysis of available experimental results for subcritical water hydrolysis studies to produce
sugars from hemicellulose and cellulose biomass fractions was conducted in order to define
the process conditions that result in high yields while limiting the formation of unwanted
decomposition products. The proposed process consists of a semibatch fractionation process in
which biomass is initially fed to the reactor and water flows continuously though the packed
bed. A first hydrolysis is carried out at 185 °C and 50 bar, in which mostly hemicellulose sugars
are hydrolyzed, followed by a second hydrolysis in which temperature is increased to 300 °C
while keeping constant all the other conditions to hydrolyze cellulose to glucose. A final step
considers supercritical water oxidation at 400 °C and using hydrogen peroxide to recover the
energy produced in the oxidation process for heat and electricity used in the process.
Although the proposed process is semicontinuous, the simulation in Aspen Plus was based on
a continuous process based on the assumption that an industrial process could be carried out in
such manner if comprised by at least three reactors in parallel, in a way that hemicellulose and
cellulose hydrolysis as well as oxidation are always taking place in a separate reactor. The
conventional biorefinery process in which acid hydrolysis is followed by enzymatic hydrolysis
was also simulated based on available information on pretreatment steps, chemical reagents,
and process conditions. An LCA analysis was conducted in OpenLCA software, in which the
goal and scope was to examine the environmental performance of the proposed process in
comparison to the conventional process and the functional unit was the input lignocellulosic
waste flow of 6500 tons to produce fermentable sugars, electricity and heat, considering a gate-
to-gate scenario. Mass and energy balances from the simulations were used for the inventory
analysis and the method Recipe midpoint was selected for impact assessment and
interpretation.
The LCA interpretation step indicated a better environmental performance of the proposed
process implementing subcritical and supercritical processing in comparison to the
conventional process, which could be attributed to a lower energy requirement, since part of
the energy supplied to heat and pressurize water and biomass to subcritical and supercritical
conditions can be recovered through heat integration and to flash depressurization that removes
water from the hydrolysates, avoiding additional water evaporation. The proposed process also
reduces waste streams generated in the neutralization of acids in the conventional process as
well as waste biomass produced in the enzymatic hydrolysis. The proposed biorefinery
integrating subcritical and supercritical processing for fractionated hydrolysis and energy
production by supercritical water oxidation could be an efficient method for waste biomass
valorization, since technologically is a simpler process with less steps than the conventional
process.

Keywords: Subcritcal hydrolysis, sugar cane waste, biorefnery, LCA.

154
 Gurguéia nut (Dipteryx lacunifera Ducke) cake as a source for
obtaining extracts rich in bioactives using pressurized liquid
extraction

Gabriela Polmanna*, Pedro Henrique Santosb, Rômulo Alves Moraisa, Sandra Regina
Salvador Ferreirab, Jane Mara Blocka*
a
Department of Food Science and Technology, Federal University of Santa Catarina, Florianópolis,
SC, Brazil.
b
Chemical and Food Engineering Department, Federal University of Santa Catarina,
Florianópolis, SC, C.P. 476, 88040-900, Brazil.
*corresponding author email address: gabipolmann@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Dipteryx lacunifera, popularly known as gurguéia nut, is a nut native to the Brazilian Cerrado
with potential for commercialization and consumption. However, the gurguéia nut is still little
known and explored. The cake obtained from the cold pressing of gurguéia nuts is rich in lipids
and bioactive compounds. The objective of this work was to study the potential for sustainable
use of gurguéia nut cake using different clean extraction methods. The gurguéia nut cake
extracts, obtained by pressurized liquid extraction (PLE) and microwave-assisted extraction
(MAE), the composition of phytochemicals, and the antioxidant potential were evaluated, using
as a strategy for the full use of this raw material. Gurguéia cakes obtained by a hydraulic press
were used to obtain the cake 1 (C1). The residual oil of C1 was extracted using two different
extraction methods for obtaining cake 2 (C2) and cake 3 (C3). C2 was obtained from C1 using

155
SFE with CO2 (flow rate was set at 1.0 kg h−1) at 40 °C, 20 MPa for 180 min; and cake 3 (C3)
was obtained from C1 by Soxhlet with hexane (a ratio of 1:30 (w/v)) at 69 °C and for 6 h. C2
and C3 were used as raw materials to obtain extracts by PLE, with 3 g of cake, at 60 °C, 100
bar, and solvent flow rate of 3 mL min–1 of ethanol (99.9%), or ethanol and water (30 or 70%,
v/v); and MAE, with 1 g of sample and 20 mL of ethanol (99.9%), or ethanol and water (30 or
70%, v/v) at 60 °C. The yield of extraction (%, g g−1), total phenolic compounds (TPC),
flavonoids, flavonoids, ortho-diphenols (ODP), condensed tannins (CT), and antioxidant
potential (ABTS, TRC, CUPRAC, FRAP, DPPH, ILP) were determined. The highest yields
were observed for C2 (39.98%, g g−1) and C3 (35.60%, g g−1) obtained by PLE with 70%
ethanol. On the other hand, the lowest yields were verified in C2 and C3 (7.82 and 9.24% g
g−1, respectively) obtained by MAE with pure ethanol. The C2 extract obtained by PLE with
30% ethanol showed a concentration of TPC (4624.74 mg GAE 100 g−1), flavonoids (418.36
mg CE 100 g−1), flavonols (150.35 mg QE 100 g−1) and ODP (381.91 mg CGA 100 g−1) higher
when compared to the other extracts. The highest phenolic contents by PB (810.79 mg GAE
100 g−1) and CT (515.03 mg CE 100 g−1) were observed for the C2 extract, obtained by PLE
with 70% ethanol. The C2 extract, obtained by PLE with 70% ethanol, had higher antioxidant
potential for ABTS (1319.08 mmol TE 100 g−1), TRC (1064.00 mg QE 100 g−1) and CUPRAC
(70.38 mmol TE 100 g−1) than the other extracts. The C3 and C2 extracts showed the highest
values for DPPH (81.40%) and FRAP (368.36 mg TE 100 g−1), respectively. Furthermore, the
C2 extracts obtained by PLE exhibited approximately 75% of ILP. These data indicated that
30 and 70% ethanol-based extracts obtained by PLE have the highest antioxidant potential for
C2 and C3 cake samples. The yield was affected by the extraction method, type of solvent, and
quality of the raw material (initial lipid content). Sample C2 had a higher content of
phytochemicals than sample C3. These results are because the extraction process with
supercritical CO2 is not operated at a high temperature as in the Soxhlet extraction process.
PLE proved to be effective for recovering bioactive compounds from cakes of gurguéia nut. In
addition, this process can retard the degradation of components recovered by oxidative
processes due to their antioxidant characteristics. The PLE temperature (60 °C) probably
increased the recovery of antioxidants from the sample or hydrolyze new compounds with
antioxidant characteristics. Likewise, heat treatment can induce the formation of new bioactive
substances. The results indicated that the extracts obtained from gurguéia cake using PLE with
ethanol and water are rich in total phenolic compounds and present high antioxidant potential.

Keywords: Brazilian nut, pressurized liquid extraction (PLE), co-products, phenolic

compounds, antioxidant potential.

156
 Scaling up of the Extraction of Dill Essential Oil
Miguel Francoa, Marleny D.A. Saldañaa,*
a
Department of Agricultural, Food and Nutritional Science, University of Alberta
116 St & 85 Ave, Edmonton, Alberta, Canada
*marleny.saldana@ualberta.ca

GRAPHICAL ABSTRACT

ABSTRACT

The utilization of supercritical CO2 (SC-CO2) for extracting essential oils has gained significant
attention in recent decades due to its high yield, quality of the extract, and absence of toxic
solvents. Essential oils, consisting of hydrocarbons and oxygenated compounds, possess
various biological properties, such as antimicrobial and antioxidative activities. Dill seeds,
Anethum graveolens, are recognized for their contents of carvone and D-limonene. This study
aimed to determine the kinetics and optimal conditions for extracting essential oil from dill
seeds at laboratory scale (~3.2 g of sample) and scale it up threefold and tenfold (~9 g and 32
g of sample). SC-CO2 extraction of the essential oil was conducted within a pressure range of
150 to 350 bar and temperatures between 40 and 60 °C for a duration of 180 min while
maintaining constant flow rates of 2.30 mL/min at lab-scale and 1.50 L/min at pilot-scale. The

157
findings revealed that the most favourable extraction conditions at lab scale were achieved at
350 bar and 50 °C, resulting in a maximum yield of 13.69%. For the pilot plant experiments,
the yield ranged between 11.88 and 12.87% at the same conditions. According to the
experiments, pressure had more significant impact on the yield than temperature. The kinetics
curve exhibited linear behaviour during the initial 150 min of operation, with the yield
remaining practically constant after 210 min. The essential oils were characterized using gas
chromatography analysis, where carvone and D-limonene were present in the samples. The
concentrations of these bioactives decreased gradually after the transition and diffusion-
controlled phases.

Keywords: Essential oil, extraction, kinetics, limonene, scale-up.

158
 Pressurized liquid extraction as a promise method to obtain
protein from broken black bean cotyledon

Renata Fialho Teixeiraa, Laís Benvenuttia, Débora de Oliveiraa, Acácio Antonio Ferreira
Zielinskia
a
Chemical and Food Engineering Department, Federal University of Santa Catarina, EQA/UFSC – PO
Box 476, CEP 88040-900, Florianópolis, SC, Brazil.
*renatafialhot@gmail.com (Renata F. Teixeira)

GRAPHICAL ABSTRACT

ABSTRACT
The increasing world population and the search for new food habits highlight the need for
alternative plant-based protein sources. Pulses, including beans, have been recognized as
healthy, protein-rich staple foods with adequate nutritional quality and techno-functional
properties. Emerging technologies, such as pressurized liquid extraction (PLE), can improve
the recovery and functionalization of bean proteins compared to conventional heating-stirring
extraction (HSE). Until now, no study has been conducted on using PLE to extract protein from
beans, despite the promising findings obtained from other plant sources. This study aimed to
evaluate the protein recovery, the in vitro protein digestibility (IVPD), and the techno-
functional properties of broken black bean protein concentrate (BBBPC) obtained by HSE and
PLE, using water and alkaline solution (pH 9.0) as solvents. BBBPC was extracted in a solid-
liquid ratio 1:20 (m/v) at a temperature of 60 °C for 15 min (PLE operated at 100 bar with a

159
flow rate of 4 mL.min-1). Then, the extracts were subjected to isoelectric protein precipitation
steps (pH adjustment to 4.5), followed by centrifugation, neutralization, and lyophilization.
IVPD analysis was performed using a multi-enzymatic method (peptidase, trypsin, and α-
chymotrypsin), and the solubility of the extracts was determined at neutral pH. Evaluated
techno-functional properties of BBBPC were: Emulsifying Activity Index (EAI), Emulsion
Stability Index (ESI), Foaming Capacity (FC), Foam Stability (FS), Oil Holding Capacity
(OHC), and Water Holding Capacity (WHC). Protein fractions obtained by the PLE (water)
and HSE (alkaline) methods showed the highest protein purity (86.74 % and 86.06 %,
respectively), while PLE (alkaline) showed the highest protein yield (23.9 %) against the 18.8-
22.2 % yield of HSE. BBBPC exhibited remarkable solubility (83.3 %) and IVPD (70.8 %)
when subjected to PLE under alkaline conditions, surpassing HSE by 26.6 % and 5.9 %,
respectively. The characteristics of foaming and emulsification are primarily related to protein
solubility, molecular structure, and hydrophilic, hydrophobic, and protein-to-protein
interactions. Consequently, the increased solubility of BBBPC (alkaline PLE) significantly
improved EAI (8.4 m²/g), ESI (31.5 min), and FC (125.6 %), with enhancements of 212.2 %,
77.5 % and 44.9 %, respectively, compared to HSE (alkaline). However, all treatments
maintained above 90 % of the initially formed foam after 60 min, demonstrating high stability.
The good foam stability is ideal for application in food products such as mousses and toppings.
In contrast, proteins with good emulsification capacity can produce hamburgers, sausages, and
other meat substitutes. OHC and WHC are associated with juiciness, tenderness, and flavor
retention. Application of PLE with both solvents resulted in the highest OHC values (11.8-
11.9 g/g), and the highest WHC (2.2 g/g) was obtained with HSE (water). A high WHC is
related to protein denaturation, breaking down weaker interactions and increasing the exposure
of hydrophilic groups interacting with water. In conclusion, using PLE (alkaline) improved the
techno-functional properties of BBBPC, attributed to the conformational changes in protein
structure, like unfolding when submitted to high-pressure fluid. Implementing PLE technology
for protein recovery holds promise for its utilization in protein supplements, meat analogs,
desserts, and bakery products, thereby enhancing the value of bean co-products. Furthermore,
this approach addresses the growing need for alternative protein sources by employing an
environmentally friendly, safe, and efficient method.

Keywords: Pulse proteins, digestibility, alternative source, emerging technology.

160
 Supercritical fluid extraction of oil from Mauritia flexuosa pulp:
a comparison with traditional extraction techniques

Marcela P. Carrillo-Bautistaa,b*, Luisa L. Orduz-Díazb, Juliana E. C. Cardona-Jaramillob,

Gustavo Bolañosa
a
Laboratory of Applied Thermodynamics and Supercritical Fluids
School of Chemical Engineering, Universidad del Valle
Calle 13 # 100-00 Bldg E30, Cali, Colombia
b
Instituto Amazónico de Investigaciones Científicas SINCHI
Calle 20 # 5-44, Bogotá, Colombia

*Corresponding author: marcela.carrillo@correounivalle.edu.co.

GRAPHICAL ABSTRACT

ABSTRACT
Canangucha (Mauritia flexuosa L.f) is a palm that belongs to the Arecaceae family and is a
valuable non-timber forest product present in large quantities in the Amazon region. This
species has a wide distribution and density in the Colombian Amazon, and it is being
considered as an option for biodiversity conservation through its sustainable use. The main
product of this plant is an antioxidant-rich oil obtained from its fruits, which has a great
potential as a natural ingredient for the cosmetic and food industries.

161
Amazonian communities usually extract the oil using hydraulic pressing (HP) because this is
a simple, environmentally friendly, and cost-effective process, although it has a limited
production capacity. On the other hand, another possible technique is expeller pressing (EP),
which is a higher-capacity technology for continuous or semicontinuous processes but requires
a fine control to avoid the loss of physicochemical properties and bioactive compounds of the
product, a situation that might be avoided using supercritical fluid extraction (SFE).
In this work, we compared these three technologies for obtaining oil from M. flexuosa fruits.
Dehydrated pulp from fruits harvested in Montañita-Caquetá (Colombia) was used as raw
material. Samples of this pulp were respectively subjected to HP at 200 ton and 30 ºC, to EP at
20 rpm and 70 ºC, and to SFE with carbon dioxide at 22.3 MPa and 42 ºC. In each case,
extraction yield and some quality parameters of the product, such as lipid profile, quality index,
density, refraction index, total carotenoids and β-carotene contents of the extracts were
determined.
The results show that the extraction yield of SFE (26.0%) was higher than those obtained for
EP and HP (19.9% and 21.0%, respectively), and the quality parameters of the oil did not
exhibit any significant variation due to the extraction process. These results indicate that SFE
is an attractive alternative for producing M. flexuosa pulp oil that not only preserves the oil’s
quality but also improves the extraction yield.

Keywords: Mauritia flexuosa, natural oil, supercritical fluid extraction, amazonian species,
non-timber forest products.

162
 Peach seed cake (Prunus persica): a comparative study of
different extraction methods for the recovery of protein fraction
Carla R.M. Rudke,, Talyta M. S. Torres, Acácio A. F. Zielinski, Sandra R.S. Ferreira*

Federal University of Santa Catarina, Department University of Santa Catarina, PO BOX 476 88040-
970, Florianópolis SC, Brazil
*corresponding author email address: s.ferreira@ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Peach fruits (Prunus persica) belong to the Rosaceae family and are widely appreciated in
flavor, texture, juiciness, and nutritional value. Nowadays, peach production and processing
represent a significant market, and it is estimated that there are currently more than 400
cultivars worldwide. Several studies have already evaluated peach by-products, with the
identification of various bioactive compounds. However, the protein recovery from peach by-
products using emerging techniques still remains unexplored. Besides, the proximal
composition showed that peach seed has oil content of 33.40%±2.0, and protein content of
29.13%±0.63. The sequential extractions consisted of supercritical fluid (SFE-CO2) for the oily
fraction recovery (yield of 29.53% ±0.45), followed by protein recovery from the defatted cake
using the following methods: heat stirred extraction-HSE, and pressurized liquid extraction-
PLE, with alkaline water as solvent. The PLE, conducted at 100 bar 40°C for 10 min, provided
the highest protein recovery by Kjeldahl method (95.10%±0.98), followed by the HSE

163
technique (55.09%±0.35). Regarding the functional properties of the protein fraction, PLE
sample showed the highest values of turbidity (94% ± 0.02), sulfhydryl content (0.47 nmol.mg.
protein-1 ± 0.01), emulsion activity (2.75 m2/g ±0.25), foaming property (16.33% ± 1.15),
hydrophobicity (slope-942), whiteness index (36.67) and isoelectric point (5.09 ± 0.57).
Therefore, it is suggested that an isolated protein with better functional properties was obtained
by PLE. The high-pressure method provided a very promising result for protein purification,
creating alternatives to using the peach seed, adding value and lining up the concepts of
upcycling.

Keywords: Supercritical fluid, by-product, emergency techniques, aminoacid, alternative

protein

164
 Recovering bioactive compounds from pineapple waste: a
comparison between pressurizes liquid and supercritical fluid
extractions
Felipe A. Maiaa,*, Luiz H. Fasolina
a
Departament of Food Engineering and Technology, Faculty of Food Engineering University of
Campinas, UNICAMP, Campinas – SP, Brazil
*corresponding author email address: fmaia780@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Pineapple is a popular fruit consumed worldwide. As a consequence of fruit processing, about
50% of the raw material becomes waste with high potential for recovery of bioactive
compounds. In this perspective, this work aimed evaluating high pressure techniques
(supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE) to recovery
bioactive compounds from pineapple waste. Pineapple waste was formed by peel and bagasse
from juice processing. The SFE extractions were performed based on a full factorial
experimental design with different levels of pressure (10, 20 and 30 MPa), temperature (40 and
60 °C) and percentage of co-solvent (5% and 15% w/w ethanol). On the other hand, PLE
extractions were evaluated at different temperature levels (60, 80 and 100 °C) and ethanol
percentage (50, 75 and 100% v/v). The extracts were evaluated for overall yield, phenolic
compounds, flavonoid, carotenoid content and antioxidant activity. Then, overall extraction
curves (OEC) were performed for the optimized conditions to understand the kinetic behavior
of the extraction. It was observed that PLE had the highest overall yield of extract (38.7% w/w),

165
phenolic compounds (9.39 ±0.56 mg GAE/g RM dw), flavonoids (4.14 ± 0.45 mg EC/g RM
dw), carotenoids (26.18 ± 1.04 µg β-carotene/g RM dw) and antioxidant activity. This behavior
occurs because PLE uses higher temperature, which increase the mass transfer coefficient. In
addition, PLE requires a shorter extraction time and, consequently, there is less degradation of
the compounds. Another possibility is that generally hydroalcoholic solutions are more
efficient in solubilizing hydrophilic compounds than pure solvents. The determination of the
global extraction curves showed that the PLE reaches the diffusional period in times less than
50 min, while the duration of the SFE was approximately 5 times longer for all compounds
evaluated. Among the phenolics evaluated the SFE was able to selectively extract ferulic acid
(60%) in the optimal extraction condition. Therefore, PLE proved to be the best technique for
extracting bioactive compounds from pineapple waste when compared to SFE. In addition, the
optimized operating condition showed that PLE is a more sustainable technique, with less time
and energy demand.

Keywords: pineapple, SFE, PLE, antioxidant activity.

166
 Integration of alternative solvents for enhanced stability of dill (Anethum
graveolens L.) aroma volatile compounds
Jelena Vladica*, Stela Jokicb, Krunoslav Aladicb, Igor Jerkovicc, Ana Rita Duartea
a
Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal
b
Faculty of Food Technology Osijek, University of Josip Juraj Strossmayer of Osijek, Osijek, Croatia
c
Faculty of Chemistry and Technology, University of Split, Split, Croatia
*corresponding author email address: jelenavladic@fct.unl.pt

GRAPHICAL ABSTRACT

ABSTRACT
The aim of this work was to establish the process of obtaining aroma volatile compounds (VCs)
with an extended shelf life that integrates the use of two alternative solvents, supercritical
carbon dioxide (SCO2) and deep eutectic solvents (DES). Dill (Anethum graveolens L.) was
used as a source of aroma organic VCs. SCO2 was applied for the recovery of volatile lipophilic
components at 40 °C and 80 bar. The isolated components were further dispersed in betaine-
based mixtures: betaine/levulinic acid (Bet/LA), betaine/glycerol (Bet/Gly), and
betaine/ethylene glycol (Bet/EG), while SCO2 dill extract was used as a control. The stability
of the samples was monitored during storage at room temperature for 6 months. The volatile
headspace compounds (%) of A. graveolans samples were extracted by headspace solid-phase
microextraction and analyzed by gas chromatography-mass spectrometry.
Alterations in the headspace profiles were detected in the samples during storage, which
corresponds to oxidation, evaporation, and other transformation reactions. The most dominant
component in the samples was carvone with 43.42 - 60.44% at the start (month 0) and its
increase was observed during 6 months in all samples. The most pronounced changes in the
headspace profile were found in the control, where the second most abundant component
limonene (39.87%) had a drop to 1.24% after 6 months. A significant decrease in limonene
was also observed in the Bet/Gly sample, from 33.85 to 3.76%, while the oscillations in the
other two DESs, Bet/LA and Bet/EG, were significantly milder (from 46.00 to 28.21% and
from 39.16 to 15.96%, respectively). Additional confirmation of the reduced stability of the
control sample was the development and percentual increase of acetic acid, which may be
responsible for changing the sensory characteristics of the product. The development of low

167
molecular weight organic VCs that cause changes in sensory characteristics and may have a
harmful effect on human health was not detected in samples stabilized in DES.
The obtained results indicate that the dispersion of dill aroma VCs recovered by SCO2 in
Bet/LA and Bet/EG significantly decelerated degradation and ensured their prolonged stability.
The established approach represents a simple solution that ensures the attainment and stability
of aroma VCs of dill, in an environmentally friendly way. Additional benefits of the process
are the more effective use of natural resources, and a safe procedure which does not require
additional equipment and does not generate solvent waste.

Acknowledgements
This project has received funding from the European Union’s Horizon 2020 research and
innovation program under the Marie Sklodowska-Curie grant agreement No 101003396.

Keywords: deep eutectic solvent, stabilization, aroma volatile compounds, supercritical carbon
dioxide

168
 NARINGIN PROCESSED BY GAS ANTISOLVENT
TECHNIQUE
Patricia V. Oliveiraa, g, Jônatas L. Diasa, Gean P. S. Aguiar b,c, Ketelin Z. Kuhnd, Otavio
Sanaiottod, Ana C. Provinellid, Carla F. Danield, Adailton Bortoluzzie, Anna M. Siebelb,d,f,
Lindomar Alberto Lering, Claudio Trapellag, Liz G. Müllerb , J. Vladimir Oliveiraa,b*
a
Department of Chemical and Food Engineering, UFSC, Florianópolis, SC, Brazil, 88040-
900
b
Graduate Program in Environmental Sciences, Community University of Chapecó Region,
Unochapecó, Chapecó, SC, Brazil
c
Brazilian Service of Support to the Micron and Small Companies, SEBRAE, Chapecó, SC,
Brazil, 89803-003
d
Health Sciences School, Community University of Chapecó Region, Unochapecó, Chapecó,
SC, Brazil
e
Department of Chemistry, UFSC, Florianópolis, SC, Brazil
f
Institute of Biological Sciences, Universidade Federal do Rio Grande, FURG, Rio Grande,
RS, Brazil
g
Departament of Chemical, Pharmaceutical and Agricultural Science, Università degli Studi
di Ferrara, UNIFE, Ferrara
*corresponding author email address: jose.vladimir@ufsc.br

GRAPHICAL ABSTRACT

169
ABSTRACT
Naringin is a natural flavanone glycoside found in Chinese herbal medicine and citrus fruits.
Studies have shown that naringin has numerous biological and pharmacological properties,
such as antioxidant, anti-inflammatory, antidepressant, anti-apoptotic, and neuroprotective
properties. Naringenin has promising biological activity but finds pharmacokinetic barriers that
make it difficult to apply pharmaceuticals. Naringin has low solubility in water (0.6502 mg.mL-
1
), low oral bioavailability and an undesirable bitter taste. In this sense, various techniques have
been used to improve the low dissolution rate, solubility and bioavailability of such bioactive
compounds. The use of supercritical fluids offers several advantages over conventional
techniques, such as the use of moderate temperatures and an inert atmosphere that prevents
product degradation, and most importantly, the avoidance or reduction of the use of organic
solvents. This study aimed to obtain microparticles of naringin by the gas antisolvent technique
(GAS). For this purpose, a central composite experimental design 24 was adopted to evaluate
the effect of concentration (5; 7.5; 10 mg.mL-1), temperature (35, 40, and 45°C), pressure (8,
10, 12 MPa), and CO2 flow rate (5, 10, and 15 mL.min-1) on process yield and particle size.
The organic solvent used in the process is acetone. A series of analyses were performed
(XRDP, FTIR, SEM and DSC) and the dissolution rate was evaluated for future in vitro/in vivo
applications. The results obtained in this work showed that after processing by the GAS
technique, naringin run 11 (5 mg.mL-1, 12 MPa, 35°C, 15 mL.min-1) exhibited amorphous
characteristics with a yield of 61.37% and an average particle size of 0.83 µm. A 19-fold
reduction in particle size was observed compared to commercial naringin (15.71 µm). The
dissolution rate of run 11 naringin particles in distilled water was about 5 times higher than that
of commercial naringin. The dissolution results are probably related to a decrease in particle
size and an increase in the specific surface area of the substance (about 5-fold), which increases
the dissolution rate, as described by the Noyes–Whitney equation.Our data suggest the
potential application of amorphous naringin, which is due to its increased dissolution rate and
enhanced bioavailability. The GAS technique shows to be a promising approach for the
pharmaceutical application of this compound.

Keywords: GAS, supercritical fluid, bioactive compound

170
 Recovery of phenolic compounds from pink pepper fruits
Evertan A. Rebelattoa, Luciano Vitalib, Adenilson R. Rudkea, Kátia S. Andradea,
Sandra R. S. Ferreiraa
a
Department of Chemical and Food Engineering, Universidade Federal de Santa
Catarina/UFSC, 88040-900, Florianópolis, SC, Brazil
b
Department of Chemistry, Federal University of Santa Catarina, Florianópolis, SC, Brazil
*corresponding author email address: etorebelatto@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Schinus terebinthifolius Raddi, Anacardiaceae family, is a Brazilian native plant, also known
as Aroeira or Brazilian pepper tree. In traditional medicine, pink pepper has been used as a
remedy for ulcers, respiratory problems, wounds, rheumatism, gout, diarrhea, skin ailments,
and arthritis, as well as to treat tumors and leprosy. Integrated high-pressure processes,
combining Supercritical CO2 extraction (SFE-CO2) at 30 MPa and 333 K, Pressurized Liquid
Extraction (PLE) at 10 MPa and 333 K, and Subcritical Water Extraction (SWE) at 10 MPa
and 403 K, were used to obtain antioxidant compounds from pink pepper fruits. The recovered
extracts were characterized in terms of total phenolic content (TPC) and antioxidant activity,
by the methods DPPH (1,1-diphenyl-2-picrylhydrazyl method), ABTS (2,2′- Azino-bis (3-
ethylbenzothiazoline-6-sulfonic acid) and FRAP (Ferric reducing antioxidant power), and the
results showed high antioxidant potential from the SWE sample (ranged from 114.16 to 818.22
μmol TE.g-1). The chemical profile of different extracts obtained from pink pepper fruits was
evaluated by HPLC for the selection of the most promising extract for future applications. The
five main compounds identified and quantified were ellagic, gallic and protocatechuic acids
and the flavonoids kaempferol and myricetin. Gallic acid, the main quantified compound,
presents anticancer, antiulceronic, antiinflammatory, antimicrobial, and antitumor activities.
PLE and SWE assisted by SFE showed to be clean, efficient, and green alternatives for the
recovery of antioxidants compounds.

Keywords: Schinus terebinthifolius, bio-refinery, high-pressure extraction, phenolic

compounds.

171
 Sequential extraction of tocols and carotenoids from Butia
capitata fruits using supercritical carbon dioxide and pressurized
fluids
Gabriel Sthefano Lourenço Pereiraa, Patrícia Tonon de Souzaa, Sara Fraga, Klicia Araujo
Sampaioa*
a
University of Campinas, Cidade Universitária Zeferino Vaz – Barão Geraldo, Campinas, Brazil
*klicias@unicamp.br

GRAPHICAL ABSTRACT

ABSTRACT
Butia capitata, also known as coquinho-azedo, is a tropical fruit tree native to the Brazilian
Cerrado, highly valued for its bioactive compounds, such as tocols and carotenoids. These
compounds contribute to the fruit's nutritional and medicinal properties, including anti-
inflammatory, cardiovascular, and potentially anti-cancer effects. Traditional extraction
methods for these bioactive compounds have limitations, prompting exploration of alternative
techniques. Sequential extraction using supercritical carbon dioxide (scCO 2) and pressurized
fluids (PFE) emerges as a promising approach for obtaining these compounds from Butia
capitata fruit pulp. The scCO2 extraction employs carbon dioxide at its critical point to
efficiently extract a wide range of compounds, including carotenoids and tocols. Subsequently,
PFE utilizes pressurized solvents such as ethanol and water, further enhancing extraction
efficiency. These alternative techniques offer advantages like selective compound extraction,

172
reduced environmental impact due to lower organic solvent use, and better preservation of
bioactive compounds under mild extraction conditions. This study employs sequential
extraction with scCO2 and PFE to obtain Butia capitata fruit pulp extracts enriched with tocols
and carotenoids.
Ripe fruits were procured from the Municipal Market in Montes Claros, Minas Gerais state,
Brazil, and experimental assays were conducted in the Laboratory of Extraction, Applied
Thermodynamics and Equilibrium (ExTrAE) at the State University of Campinas
(UNICAMP). The fruit pulps were separated using a fruit pulper, followed by lyophilization
for drying and knife milling for grinding. The supercritical extraction unit involved an extractor
containing packed raw material, forming a particle bed supplemented with glass beads. The
scCO2 was conducted at a flow rate of 1.5 L/min, and the extract was collected in a collector
with controlled flow, temperature, and pressure. The sequential fixed-bed extraction method
was employed after prior studies, and it demonstrated improved oil extraction yields. For scCO2
extraction, the process was conducted at a fixed temperature of 50 °C and 350 bar, resulting in
supercritical carbon extract (SCE). Subsequently, the extraction residue underwent pressurized
ethanol and water extraction at 150 bar, yielding the sequential ethanol extract (SEE) and
sequential water extract (SWE), respectively. Each stage lasted 5 hours. The yield of
extractions was assessed, total carotenoids quantified using spectrophotometry, and tocols
profiles determined via ultra-high performance liquid chromatography with mass spectrometer
(UHPLC-MS). Experimental data were statistically compared using the Tukey test at a 5%
significance level, employing R® software.
The sequential method exhibited a high cumulative yield of 68.78% (26.32 ± 1.14% for SCE,
28.70 ± 1.9% for SEE, and 13.8 ± 0.8% for SWE). Regarding total carotenoids, SCE extract
demonstrated a significantly higher concentration of 56.14 ± 1.32 µg/g, followed by SWE with
35.44 ± 6.69 µg/g and SWE with 0.1 ± 0.1 µg/g. Analysis of tocols composition revealed that
the SCE extract contained 8.6 ± 0.1 µg/g of β/γ-tocopherol, 8.5 ± 0.1 µg/g of α-tocopherol, 0.3
± 0.1 µg/g of δ-tocopherol, and 1.6 ± 0.1 µg/g of β/γ-tocotrienol. The SEE extract displayed
11.9 ± 0.3 µg/g of β/γ-tocopherol, 11.8 ± 0.2 µg/g of α-tocopherol, 0.9 ± 0.1 µg/g of δ-
tocopherol, and 1.4 ± 0.1 µg/g of β/γ-tocotrienol. In contrast, SWE extract exhibited a low
concentration, containing only 1.6 ± 0.1 µg/g of α-tocopherol. The diminished content of
compounds found in SWE extract may result from water's inefficiency as a solvent for
lipophilic compounds. However, it's also important to consider that the extract might be
exhausted after SCE and SEE stages.
In conclusion, this study highlights the potential of sequential extraction using scCO2 and PFE
as a promising method for obtaining bioactive compounds, including tocols and carotenoids,
from Butia capitata fruit pulp. The sequential method yielded a cumulative high extraction
efficiency, with substantial carotenoid and tocols extraction. The SCE extract, obtained after
scCO2 extraction, exhibited the highest levels of total carotenoids and tocols. SWE extracts
displayed a lower content of these compounds, potentially due to water's inherent inefficiency
in extracting lipophilic compounds and the exhaustion of the raw material. The primary tocols
extracted were β/γ-tocopherol and α-tocopherol, with their amounts varying significantly
between extracts. The application of sequential extraction with scCO2 and PFE offers
numerous advantages, including selectivity, reduced environmental impact, and better
compound preservation. Further investigations could explore optimized extraction conditions
and potential health benefits linked to consuming these enriched extracts.

Keywords: Cerrado, Coquinho-azedo, Sequential extraction.

173
Recovery of extracts rich in phenolic compounds from buritirana
pulp (Mauritella armata) using high-pressure extraction processes

Rômulo Alves Morais 1*; Evertan Antonio Rebelatto 2; Sandra Regina Salvador Ferreira2;
Jane Mara Block 1
1
Graduate Program in Food Science, Department of Food Science and Technology, Federal
University of Santa Catarina (UFSC), Florianopolis 88034-001, Brazil.
2
Chemical and Food Engineering Department, Federal University of Santa Catarina (UFSC),
Florianópolis 88040-900, Brazil.
*corresponding author email address: romuloitallo2505@gmail.com

GRAPHICAL ABSTRACT

174
ABSTRACT

Buritirana (Mauritella armata Mart.), a fruit of the Arecaceae family native to the Brazilian
Cerrado, is rich in lipids and bioactive compounds with high antioxidant potential. They are
widely consumed in the northern states of Brazil in natura or as juices, sweets, jellies, and
fermented drinks. In this work conventional and emerging extractions of bioactive compounds
from buritirana pulp were studied. The extracts were obtained by Soxhlet (SOX) and by
pressurized liquid extraction (PLE) using 70% ethanol and 30% water as solvent at 10 MPa,
60 ºC, and the flow rate at 4 mL min−1 for 50 min. The conditions used in the extraction process
were established by previous research carried out by the research group with matrices similar
to the present study. Total phenolics, total flavonoids and flavonols and ortho-diphenols and
antioxidant potential (DPPH, FRAP, ABTS and CUPRAC). Total phenolics, flavonoids, and
total flavonols and ortho-diphenols, and antioxidant potential (DPPH, FRAP, ABTS, and
CUPRAC) were determined in the obtained extracts. The yield obtained from Soxhlet method
was 17.67 % and for PLE 35.76 %. The phytochemical analysis showed that the extraction
using Soxhlet method substantially affected the amount of the bioactive compounds in the
buritirana pulp. The total phenolic, flavonoids and flavonols content were between 744.48 mg
AGE 100 g−1 and 533.87 mg AGE 100 g−1; 437.98 and 309.31 mg CE 100 g−1; and 181.86 and
77,69 mg QE 100 g−1, for PLE and Soxhlet, respectively. On the other hand, the content of
ortho-diphenols presented higher amount for Soxhlet when compared to PLE (572.10 and
373.59 mg CAE 100 g−1, respectively). The antioxidant potential of the recovered samples also
showed a strong influence of the extraction method. DPPH values were 60.30 % inhibition for
PLE sample, and 45.91 % inhibition for Soxhlet extract. The same behavior was observed for
the ABTS and FRAP analysis (6.74 and 3.57 mmol TE 100 g−1; 477.27 and 214.36 mg AAE
100 g−1, respectively). On the other hand, the CUPRAC method showed a different behavior,
where the extract obtained by PLE showed lower results when compared to Soxhlet from 16.14
to 12.33 mmol TE 100 g−1. The results indicated that the extraction process affects significantly
the bioactive compounds' content and the antioxidant potential of the extracts, with good
behavior presented by the PLE method. The obtained extracts can be used as alternative
ingredients in food and/or cosmetics formulation.

175
 Production of vitamin C-vitamin B3 cocrystals by the gaseous
antisolvent (GAS) technique
Clóvis A. Balbinot Filho*, Jônatas L. Dias, Evertan A. Rebelatto, Sandra R. S. Ferreira,
Marcelo Lanza

Universidade Federal de Santa Catarina, Department of Chemical and Food Engineering, PO Box
476, 88040-900, Florianópolis-SC, Brazil
*corresponding author: clovis.filho@posgrad.ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Vitamins are essential nutrients with specific physiological functions. L-ascorbic acid (Vitamin
C, VitC) and nicotinamide (Vitamin B3, Nic) are crucial in maintaining health. VitC prevents
scurvy, promotes tissue repair, and possesses strong antioxidant properties associated with
cancer prevention. Nic is an anti-pellagra vitamin and is effective in treating neurodegenerative
diseases. The development of multivitamin cocrystals, organized crystal structures formed by
molecules with complementary hydrogen bonding sites, offers a new approach for delivering
drug and nutrient compounds in pharmaceutical and food applications. In this context,
cocrystallization employing pressurized carbon dioxide (CO2) as an antisolvent stands out for
unique morphology and physicochemical properties of the the resulting powders. This study
explored the production of VitC-Nic cocrystals by the gaseous antisolvent (GAS) using ethanol
as solvent. Initially, GAS process was conducted at 45 °C, 90 bar, and CO2 flow of 10 mL/min.
During GAS process, small and voluminous cocrystal particles are precipitated inside the
chamber as the CO2 drags out the ethanol that solubilizes the solutes. The same cocrystals were

176
obtained through conventional liquid-assisted grinding (LAG) by maceration for 10 min of a
1:1 molar rate, adding a small volume of ethanol (200 uL). The obtained powders were
characterized by diffractive powder X-ray (DRX) and differential scanning calorimetry (DSC),
and the resulting spectra were compared with pure compounds and physical mixture to confirm
cocrystal formation. A Box-Behnken experimental design (15 runs) was employed to evaluate
the following GAS process variables: temperature (35, 45, and 55 °C), CO2 flow rate (5, 10,
and 15 mL/min), and the molar ratio of VitC:Nic (1:2; 1:1 and 2:1). The mass yield, the
antioxidant activity (AA), determined by the DPPH method and the particle size of the obtained
powder measured through Scanning Electronic Microscopy (SEM) images of each run were
used as the responses. DRX diffraction peaks of the cocrystals and parent compounds showed
peaks of higher intensity at angles (2θ) that coincided with pure standards in the literature. DSC
thermograms of pure VitC and Nic showed endothermic events at 195 and 130 °C, respectively,
regarding their melting temperatures (Tm). The VitC-Nic cocrystal had an intermediary Tm of
approximately 145 °C, which differs from the mixture's euthetic point, thus indicating that a
new structure has been formed. GAS cocrystals presented a varying yield (30,35-73,85 %)
which was only negatively affected (p<0.05) by temperature. The AA varied from 1.76-4.83
mmol Trolox Equivalents (TE), which means that cocrystals could retain the AA of pure
Vitamin C (7.85 mmol TE), highly affected by the molar ratio between VitC and Nic, i.e., the
VitC content. From SEM images (2,500x magnification), particle length ranged from 4.05-
12.78 μm, presenting mainly plate and needle morphologies. DRX spectra indicated that two
distinct polymorphs were obtained (Form I, when using 1:1 or 1:2 mixtures, and Form II,
when using a 2:1 proportion), independently of the other variables. Further, the experimental
design was optimized to render higher yield and AA with smaller particle sizes using the
desirability function of Statistic 12 software. At the optimum point (40 °C, 7.5 ml/min, and 2:1
molar ratio), VitC-Nic cocrystals showed a 60.25±2.16 % yield and 4.92± 0.56 mmol TE,
which is close to the predicted values by the desirability function (67.98% and 5.01 mmol TE),
and validates the GAS experiments. The scalability of the process was verified by doubling the
feed solution and increasing the scale from 1 to 6 mmol of each starting compound, resulting
in yield increments up to 86,24%. GAS is an attractive, reproducible, and scalable method to
produce solvent-free vitamin cocrystals with improved properties that might exert beneficial
health effects when consumed. Compared to conventional (LAG) techniques with few
controllable variables, GAS-produced cocrystals can modulate their properties to achieve a
specific purpose by changing operational processes. Nonetheless, the product obtained in this
study can serve as a model role for other vitamin-vitamin food-grade cocrystals.

Keywords: Cocrystallization, vitamin-vitamin cocrystals, supercritical carbon dioxide,

antioxidant activity, yield.

177
 High-pressure phase equilibrium data for carbon dioxide + 2-
methyltetrahydrofuran-3-one and carbon dioxide + 2-
methyltetrahydrofuran-3-one + glucose systems
Eulália L. S. Barrosa,*, Evertan A. Rebelattoa,*, Diego A. Mayera,b
J. Vladimir Oliveiraa
a
Department of Chemical and Food Engineering, Federal University of Santa Catarina, EQA/UFSC-
PO Box 476, 88040-900, Florianópolis, SC, Brazil
b
Departamento de Engenharia de Petróleo, Centro de Educação Superior da Foz do Itajaí/CESFI,
Universidade do Estado de Santa Catarina/UDESC, 88336-275, Balneário Camboriú, SC, Brazil.
*barroseulalialopes@gmail.com (Eulália L. S. Barros)

GRAPHICAL ABSTRACT

ABSTRACT
Sugar esters are surfactants with strong emulsifying, dispersing, stabilizing, and detergent
properties. However, its production remains challenging for researchers due to problems
related to the homogenization of substrates, especially sugar. In addition, production of sugar
esters most often occurs by traditional chemical esterification, which does not meet the criteria
related to efficiency and sustainability. In this way, the development of strategies to increase

178
sugar solubility in more efficient and sustainable processes has been proposed. As an
alternative, enzymatic esterification reactions in pressurized fluids have been studied to
overcome some disadvantages of conventional methods. In this process, it is important to
choose the appropriate operational conditions to facilitate the solubility of the substrates.
Therefore, the objective of this work was to investigate the phase equilibrium data for the
binary (CO2 + 2-methyltetrahydrofuran-3-one) and ternary (CO2 + 2-methyltetrahydrofuran-3-
one + glucose) to distinguish regions where a solid phase occurs and establishing operational
conditions for synthesizing sugar esters at high pressures. The experiments were performed
using the synthetic visual method over the temperature range of 313.15 to 343.15 K and
pressures up to 13.06 MPa. For the ternary systems, two solutions were prepared with 0.0265
and 0.0530 molar% of glucose concentrations in MeTHF-3-one. Liquid-vapor phase transitions
(bubble or dew points) were observed in the presence or absence of a solid phase. Regarding
the ternary system, the increase in glucose concentration anticipated the appearance of a solid
phase for all temperatures at lower CO2 mole fractions, from 0.9249 to 0.9001, ranging from
0.0265 to 0.0530 molar% of glucose into 2-methyltetrahydrofuran-3-one. The presence of
solids did not change the liquid-vapor equilibrium behavior of the binary system about the
phase transition pressures. The Peng-Robinson equation of state with the classic quadratic
mixing rule (PR-vdW2) was used to adjust the experimental data, providing a satisfactory
representation of the equilibrium data. Through the Px diagram, the behavior of the LCST, that
is, the increase of the phase transition pressures with the temperature, was observed for all the
systems studied.

Keywords: Phase equilibrium, Supercritical fluids, Bio-based Solvent, Sugar esters.

179
 Obtaining and characterization of quinoa starch (Chenopodium
quinoa Willd.) defatted by supercritical fluids for its application
as biopolymer.
Yvonne Aliagaa, Luis Olivera-Montenegroa*
a
Grupo de Investigación en Bioprocesos y conversión de la biomasa, Universidad San Ignacio de
Loyola, Avenida la Fontana N° 550, Lima, Perú
*corresponding author email address: lolivera@usil.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
The supercritical fluids technology is an ideal method for obtaining and using all quinoa
byproducts such as oil, protein, and starch; main components of quinoa, mainly due to its
multiple advantages over other conventional methodologies based on the use of organic
solvents, so that the products obtained have no solvent residues, short processing times, high
yields and low temperatures that reduce the thermal degradation of molecules of interest. On
the other hand, starch is the major component of quinoa flour, so the objective of this study
was to obtain and characterize starch from quinoa flour of the Pasankalla variety defatted by
Supercritical to evaluate its potential use as a biopolymer in the manufacture of biodegradable
packaging. The Pasankalla variety quinoa flour was defatted using supercritical CO2 (SFE) and
a pilot level multisolvent extraction equipment, the following parameters were used:
temperature of 45°C and a pressure of 40 MPa; the CO2 mass flow was 45 g/min, the extraction
time was 240 min, and the particle size was 0.5 mm, also the defatting was evaluated using
petroleum ether (CSE).

180
The oil extraction yield, paste properties by rapid visco analyzer and mechanical properties
such as strength, elongation to failure percentage by ASTM D 882 standard test method were
evaluated. The oil yield was 90% with respect to the initial raw material content (6.4 %) in SFE
and 72.5 % in CSE, the paste temperature of in SFE was slightly higher by 0.5 % compared to
CSE, the strength and elongation to failure percentage slightly higher in SFE by 10 % and 7.2
%, respectively to CSE. These results show the superiority of the use of supercritical CO2 in
performance with respect to oil recovery and superior mechanical properties of quinoa starch,
increasing the possibility of being used as a biopolymer in the production of biodegradable
packaging and maximum utilization of by-products free of organic solvents.

Keywords: Supercritical fluids Oil extraction, quinoa starch, mechanical properties,

bioplastics.

181
 Gelatin-siloxane xerogel and criogel loaded with ibuprofen by
supercritical CO2 assisted impregnation.
Ulisses Silva do Nascimentoa, Lígia Passos Maia-Obia*
a
Federal University of ABC, Avenida dos Estados, 5001, Santo André – SP, Brazil
*corresponding author email address: ligia.maia@ufabc.edu.br

GRAPHICAL ABSTRACT

ABSTRACT
Collagen has attracted interest as a biomaterial due to its biocompatibility, degradability, and,
in the form of gelatin, its ability to form hydrogels, aerogels, cryogels, and xerogels. These
materials can serve as scaffolds for cell growth and have potential to be used as composites for
bone regeneration or on the surface of ceramic or metallic implants to enhance their
biocompatibility. Gelatin hydrogels have already been tested for impregnation and controlled
release of drugs, however, the use of supercritical CO2 as impregnation media offers great
benefits by avoiding the use of organic solvents, thus obtaining a clean product. Ibuprofen
(IBU) is an anti-inflammatory drug, a crucial class of drugs needed in implants procedures and
by impregnating it in gelatin gels, a multifunctional material can be obtained that not only
serves as a biomaterial but also has potential to release the drug in a controlled manner. The
present study aims to impregnate gelatin-siloxane xerogels and cryogels with IBU using
supercritical CO2 and evaluate the impact of the process conditions and morphology of the gels
on the load of the drug. The gels were obtained by reacting gelatin with 3-glycidyloxypropyl
trimethoxysilane (GPTMS) in aqueous media and drying the system to generate a crosslinked
structure. The drying methods were (i) ambient pressure drying in an oven to obtain a xerogel,
in the form of a film, and (ii) freeze-drying, to obtain a cryogel. The xerogel and the cryogel

182
were impregnated with IBU in a 10 mL high-pressure stainless-steel cell, in the presence and
absence of 10 % v/v of ethanol as cosolvent at 35 °C and 150 and 250 bar for 24 h, in triplicate.
Enough IBU was placed inside the cell to ensure the saturation of the CO 2 solution and the
depressurization was made by cooling the cell with liquid nitrogen before opening the exit
valve, it took aprox. 10 minutes and its rate was not controlled. The crosslinking was
characterized by gel percentage (88 % and 86 %, for the xerogel and cryogel respectively) and
by FTIR, which showed a band at 1086 cm-1 for the materials, corresponding to the stretching
vibrations of the Si-O-Si crosslinking bonds, indicating that water removal during freeze drying
and the slow water removal in the oven are efficient in promoting the crosslinking. The cryogel
was obtained as a porous material while the xerogel not, its porosity was characterized by
optical microscopy (OM), showing pores of average size of 221 µm and walls with an average
thickness of 17 µm. The supercritical processes resulted in the following loads in % regarding
the weight of impregnated IBU and the final materials: for xerogels and cryogels respectively,
2.6 ± 1.3 % and 4.4 ± 0.0 % (replicates not considered due to problems with their samples
during the process) at 150 bar without cosolvent, 2.1 ± 0.5 % and 6.0 ± 0.7 % at 150 bar with
cosolvent, 2.0 ± 1.6% and 3.8 ± 1.3% at 250 bar without cosolvent, 0.3 ± 0.4 % and 15.8 ± 2.2
% at 250 bar with cosolvent. It is seen that the impregnation is higher for the cryogels showing
that there is an additional mechanism of impregnation compared with the xerogel, in which
impregnation occurs by the diffusion of IBU into the gelatin matrix. For the cryogels, the
porous structure plays a significant role on the process, indeed, OM showed that IBU crystals
were formed inside their pores. The condition of 250 bar presented a significant higher
impregnation when cosolvent was used, which can be explained by the fact that increasing the
pressure, and also the presence of cosolvent, increase the solubility of the IBU in the scCO 2
solution, the solution permeates the porous structure and IBU precipitates inside the pores when
the pressure decreases. In the other hand, for xerogels, the impregnation decreases when the
solubility of IBU increases in the system, showing that CO2 presents a low diffusion in the
bulky matrix. FTIR of the cryogels showed the presence of the characteristic bands of the IBU,
780 and 936 cm-1, attributed to the out of plane deformation vibration of aromatic C-H and of
the aliphatic C-H, respectively, and 1686 cm-1 attributed to the stretching of the bonds O-C=O
and to the bending of the bond O-H. In this work, gelatin-siloxane cryogel was successfully
impregnated by supercritical impregnation and its morphology played a crucial role in its load,
showing that even with no considerable impregnation withing the matrix, a multifunctional
material can be formed in scCO2.

Keywords: supercritical impregnation, gelatin, cryogel, xerogel, ibuprofen

183
 Sequential supercritical extraction of extract of L. rivularis stems
using CO2 and ethanol-modified CO2
Claudia Marillána ; Edgar Uquichea*.

a
University of La Frontera 1: Claudia Marillán Baez, Av. Francisco Salazar 01145, Temuco, Chile
*corresponding author email address: edgar.uquiche@ufrontera.cl

GRAPHICAL ABSTRACT

ABSTRACT
Leptocarpha rivularis ("palo negro") is a shrub native to southern Chile. It belongs to
the Asteraceae family and treats diseases like cancer and diabetes.
It is characterized by having various bioactive compounds, such as flavonoids, terpenes, and
alkaloids, which are responsible for the antioxidant activity of the plant. Obtaining bioactive
compounds from L. rivularis stems can be done through sequential supercritical extraction, an
extraction technique based on consecutive stages with a gradual increase in the polarity of the
solvent, either by increasing the pressure or adding a cosolvent. This technique increases the
extraction yield and selectively separates the compounds for each stage. In addition, it allows
the vegetable matrix to be used until it is exhausted. In general, the extraction time of each step
is established until reaching a constant value in the accumulated extraction curve. Cumulative
extraction curves are used to obtain relevant mass transfer parameters through mathematical
models.

184
The objective was to study the sequential supercritical extraction with CO 2 and CO2 modified
with ethanol to obtain fractions of extracts characterized by their content of bioactive
compounds and to obtain mass transfer parameters by modeling the accumulated extraction
curves.
The extracts were obtained using the Spe-ed 2 supercritical extractor. 12 grams of the substrate
was used in a 50 cm3 extraction vessel. The sequential supercritical extraction was carried out
in three stages at a temperature of 60°C. Stage 1: 25 MPa con CO2 pure, stage 2: 50 MPa con
CO2 pure, and stage 3: 50 MPa and CO2 + ethanol (5%wt). The ethanol was pumped to the
CO2 feed line using an HPLC pump. Under the selected conditions, extracts were collected
periodically in glass vials to obtain the accumulated extraction curves that were later modeled
with the Fick equation, using the effective diffusion coefficient (De) as the adjustment
parameter. On the other hand, terpenoids, flavonoids, and alkaloids were identified by GC-FID
and HPLC-DAD.
The model was observed to be a good fit for the accumulated extraction curves. When studying
the coefficient of effective diffusion between the stages, it resulted in a higher De (63.3±6.36
m2/s) in stage 3 at a condition of 60 °C, 50 MPa, and CO2 + ethanol (5%wt); on the contrary,
stage 2 provided a lower De (6.37 ± 0.35 m2/s) at a condition of 60°C and 50 MPa.
Regarding the individual content of flavonoids, it was found: resveratrol, kaempferol,
myricetin, and catechin; while when analyzing the range of unique terpenes, only
caryophyllene oxide was found in the three sequential stages, and the individual content of
alkaloids, only caffeine was found for the three extraction stages.
The results showed that sequential supercritical extraction allows obtaining stem extracts with
different compositions of bioactive compounds depending on the conditions to which the
sample was subjected.

Keywords : L. rivularis stem, bioactive compounds, sequential supercritical extraction.

Thanks:
Work financed by the National Research and Development Agency of Chile through the
National Doctoral Scholarship N° 21201441.

185
Enzymatic copolymerization of ω-pentadecalactone and globalide
in supercritical carbon dioxide
Roseny D. Santosa*, Evertan A. Rebelattoa, Diego A. Mayera, J. Vladimir de Oliveiraa
a
Department of Chemical Engineering and Food Engineering, Universidade Federal de Santa
Catarina, P.O. Box 476, 88040-900, Florianópolis, SC, Brazil

*corresponding author email address: rosesantosmail@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Pentadecalactone (PDL) and globalide (Gl) are cyclic esters, which contain a double bond
susceptible to hydrolysis, i.e. biodegradation. Conventionally, polymerizations of those
lactones are carried out using organic solvents and organometallic catalysts, producing
polyesters largely used in the biomedical field. However, these methods cause a negative
impact on the environment, producing toxic residues, and additional steps of purification of the
polymer. The implementation of supercritical carbon dioxide is essential for including polymer
synthesis in the new circular economy model. Furthermore, enzymatic catalysts combine mild
reaction conditions, and enzyme reuse. In this context, this work investigates the synthesis of
the new copolymer ω-pentadecalactone-co-globalide (PDLGl) using immobilized lipase

186
Novozym®435 as catalyst in supercritical carbon dioxide. The polymerizations were carried
out in a high-pressure variable-volume reactor equipped with a sapphire window for visual
observation and a movable piston, which allows the control of the pressure in the system. Under
fixed conditions of temperature and pressure, we evaluated the influence of comonomers ratios,
different carbon dioxide densities, and the use of cosolvent in various proportions in the
polymerization media. Copolymers with different feed ratios were synthetized successfully for
all studied conditions. The good performance of the copolymerization is shown by the
calculated polymer yield, which ranged from 53.72% up to 73.98% for polymerization without
cosolvent, reaching higher yield in reactions performed in higher monomers concentrations.
When chloroform was used as cosolvent, the highest yield (73.61%) was reached with higher
concentration of cosolvent. Visually, PDLGl presented different characteristics according to
the proportion of PDL and Gl. The ones containing more PDL presented as a white powder,
while the copolymers with higher amount of Gl presented as a white hard material, inferring
large possibilities of customization in the mechanical properties of the material. These
preliminary results show the potential of a new synthetic route for a novel copolymer from the
polyester group, which enable the expansion of applications of polymeric biomaterials in the
market.

Keywords: pentadecalactone, globalide, supercritical carbon dioxide.

187
 Supercritical fluid extraction of coffee oils for
sunscreen formulations
Natalia Trullo, Isabel Mejía, Gustavo Bolaños*

Laboratory of Applied Thermodynamics and Supercritical Fluids

School of Chemical Engineering, Universidad del Valle
CL 13 100-00 Edificio E30, Cali, Colombia
*corresponding author: gustavo.bolanos@correounivalle.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
Oils present in green coffee beans and spent coffee grounds contain valuable compounds with
benefits for human health, among many as agents for skin protection against solar radiation,
due to their ability to absorb ultraviolet radiation. As natural products, they have been highly
appreciated by the cosmetics industry due to their potential to replace synthetic chemicals in
sunscreen formulations.
In this work, two low-value raw materials were selected as source of coffee oils: defective
green coffee beans (GCB), which are commonly rejected in the selection process that takes
place in a coffee farm; and spent coffee grounds (SCG), which are a byproduct of the brewing
process that is performed in coffee shops. These materials are usually considered a waste, are
produced in large quantities, and their disposition has environmental issues.

188
Samples of these raw materials were dried and subjected to cryogenic milling to an average
particle size between 0.50 and 0.84 mm. The samples were then stored in a dark and oxygen-
free environment before extraction with supercritical CO2. Then, supercritical extractions were
carried out at 50 ºC and a CO2 density of 0.9 g/mL. The extraction yields obtained were 9.3%
for SCG and 7.1% for GCB.
The extracted oils were characterized by determining their antioxidant capacity and solar
protection factor (SPF). The antioxidant capacity was determined by following the well-
established 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical methodology and was reported
as the dose of oil that halves the concentration of DPPH (EC50). The results show that the GCB
oil is 2.3 times more potent antioxidant than SCG oil. The solar protection factor was
determined by UV-vis spectrophotometry and was very similar for both oils (3.78 for SCG and
3.75 for GCB).
To establish the effectiveness of these oils in sunscreen formulations, a comparison of the SPF
of a formulation with and without the oils was made. The formulation was prepared as a cold
mixture of a base cream (ProblendÒ, Protécnica Ingeniería, Colombia) and conventional filters
such as ZnO2 and TiO2. It was found that a formulation that contains 24.5% SCG oil and 10.5%
GCB oil results in an increase in SPF from 1.03 to 30.75, a typical value for commercially
available sunscreens.
Preliminary stability tests of this formulation indicate that there was not phase separation at 5
ºC and 50 ºC for 24 hours. It was also observed that the formulation has a rapid skin absorption
and does not leave a greasy feeling. These results are promising for the recovery and
valorization of coffee processing byproducts that otherwise would become a waste whose
disposition usually causes environmental pollution.

Keywords: Spent coffee grounds, green coffee beans, coffee oil, supercritical extraction,
suncreen

189
 Gelatin-Siloxane Aerogels by Supercritical Drying
Bianca Andrade de Camposa, Lígia Passos Maia-Obia,*

a
Federal University of ABC, Avenida dos Estados, 5001, Santo André – SP, Brazil.
*corresponding author email address: ligia.maia@ufabc.edu.br

GRAPHICAL ABSTRACT

ABSTRACT
Supercritical drying is a process that is calling great attention for the obtaining of structured
porous materials for biomedical and pharmaceutical applications, i.e., the impregnation and
controlled release of active molecules. Gelatin is biocompatible and degradable material that
can form hydrogels, aerogels, cryogels, and xerogels, that are stable when crosslinked. These
materials can serve as scaffolds for cell growth and have potential to be used as composites for
bone regeneration or on the surface of ceramic or metallic implants to enhance their
biocompatibility. Supercritical drying can generate porous morphologies that differ from the
ones obtained through other drying techniques, with potential to achieving low densities and
high surface areas. The present study aims to obtain aerogels trough CO2 supercritical drying
and evaluate the impact of process conditions on the drying efficiency. To synthesize the
aerogels, gelatin was reacted with 3-glycidyloxypropyl trimethoxysilane (GPTMS) in aqueous
media and cured in a hermetically closed system in an oven to prevent water evaporation during
the process. The water was exchanged by ethanol at room temperature by placing the samples
in water/ethanol mixtures with an increasing concentration of ethanol (10, 30, 50, 70, 90 and
100 %v/v) for one hour each and the samples were stored in pure ethanol. The samples were
placed inside a 10 mL high-pressure stainless-steel cell that was heated and pressurized for the

190
supercritical drying that was done in the following conditions: {40 °C and 100 bar} and {60
°C and 200 bar} for 1 and 24 h, in triplicate, followed by a flow of around 4 g.min-1 of CO2 for
5 minutes at 75 bar and at the experimental temperature. The samples were obtained as hard
materials and FTIR of these materials show a band at 1086 cm-1, corresponding to the stretching
vibrations of the Si-O-Si bonds, indicating that a crosslinked material is formed. The drying
efficiency was determined gravimetrically, as % in weight of ethanol removal, and presented
the following values for 1 and 24 h respectively: 38 ± 6 and 52 ± 17 %weight at {40 °C and
100 bar} and 43 ± 1.9 and 99 ± 5 %weight at {60 °C and 200 bar}. The morphology at
microscale was evaluated through optical microscopy (OM) with cryofractured samples, which
showed that their surfaces of the samples presented a transparent layer and the bulky regions
presented aspects ranging from translucid to opaque, as the drying efficiency increased. This
type of material is known to be amorphous and when an opaque aspect is seen, it can be
attributed to porosity. It happens because the light suffers refraction at the interface of the pores,
therefore, the transparent layer indicates that the surface do not present significant porosity,
however it did not prevent the ethanol to be removed from the samples. Moreover, the presence
of ethanol into the bulk reduces the refraction of the light and the opacity. Pores could be
observed for the dried sample up to 200x of amplification, indicating that they are smaller than
500 nm. The drying efficiency increased for {200 bar and 60 °C}, which can be attributed to
the increasing of the vapor pressure of the ethanol as the temperature increases, favoring its
solubilization in CO2 and remotion. Also, longer processing times also increased the drying
since this process involves diffusion processes, which are time dependent. Gelatin-siloxane
aerogels were successfully obtained by supercritical drying. A further kinetic study of drying
and exploration of process parameters have the potential to reduce the drying time and to
modulate the aerogel morphology.

Keywords: supercritical drying, gelatin, aerogel

191
 Value-added Products from Wheat Straw using Pressurized
Water + Ethanol Mixtures
Ana-Xochitl Vidrio-Sahaguna, Marleny D.A. Saldañaa*
a
Department of Agriculture, Food and Nutritional Science
University of Alberta, Edmonton, T6G 2R3, AB, Canada
* marleny.saldana@ualberta.ca
GRAPHICAL ABSTRACT

ABSTRACT
Cellulose, the most abundant polymer on the earth, is the main compound of wheat straw with
approximately 32%. This compound and its derivatives like cellulose nanofibers are promising
materials for various applications. However, to date, corrosive and toxic solvents have
commonly been implemented in the isolation of cellulose. The main objective of this study was
to evaluate the effect of pressurized water + ethanol mixtures at different concentrations at
optimum processing conditions of temperature, time, and volumetric flowrate to hydrolyze
non-cellulosic material from wheat straw. Pressurized aqueous ethanol (PAE) treatment of
wheat straw was performed in a semi-continuous flow type subcritical fluid system. The
experiments were conducted at 180°C, 50 bar, and ethanol concentrations of 0–100% (v/v) for
40 min. Total sugars and total phenolic contents in the hydrolysates were analyzed according
to the phenol-sulfuric acid and Folin Ciocalteu methodologies, respectively. SCW and
pressurized 20% ethanol treatments significantly reduced wheat straw's ash, crude protein, and
fat contents. These hydrolysis treatments also showed the highest carbohydrate content,
followed by 60% and 100% ethanol treatments, which may be due to the higher proportion of
water in the solution media. Thus, more H+ and OH- ions were produced, increasing the
structural carbohydrates hydrolysis, and obtaining higher quantities of monosaccharides in the
hydrolysates. Total phenolics obtained from the hydrolysis treatment indicated the rupture of
lignocellulosic complexes. Samples treated with pressurized water + ethanol at different
concentrations showed no significant difference in phenolic content expressed as gallic acid
equivalents. In conclusion, pressurized fluid treatments successfully produced hydrolyzates
with value-added products such as sugars and phenolic compounds.

Keywords: Subcritical-water, ethanol, wheat, cellulose, hydrolysis.

192
 Multiphase Equilibria Modeling of CO2
the Homologous Series of Carboxylic Acids
Jacqueline M. Ortega Bacichetia,b, Vladimir Ferreira Cabrala,b, Francisco A. Sánchezc,d*.
a
Department of Food Engineering, Universidade Estadual de Maringá – UEM, Av. Colombo, 5790,
Bloco 13, Maringá-PR, 87020-900, Brazil.
b Chemical Engineering Program, Universidade Estadual de Maringá – UEM, Av. Colombo,
5790, Bloco E46, Maringá-PR, 87020-900, Brazil.
c
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca, (8000), Argentina
d
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253, Bahía
Blanca, Argentina
*corresponding author email address: fsanchez@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT

Supercritical fluid extraction (SFE) is an attractive substitute for conventional solvent extrac-
tion methods for obtaining fine chemicals, such as fatty and dicarboxylic acids. Carbon diox-
ide (CO2) is the most commonly used supercritical solvent due to its favorable properties in-
cluding non-flammability, non-toxicity, non-polluting nature and easy recoverability. Design-
ing efficient separation units for SFE necessitates careful thermodynamic modeling.
The knowledge of the critical properties and phase equilibria of systems involving
CO2 and carboxylic acids is crucial in the context of SFE, such as oil deacidification process-
es and extraction of acids from aqueous substrates. Nevertheless, modeling phase equilibria
in mixtures containing CO2 and carboxylic acids is a challenge because of their highly
asymmetrical behavior. Moreover, while experimental data of CO2 + monocarboxylic acids

193
are available, data of CO2 + dicarboxylic acids are scarce. Consequently, the development of
predictive thermodynamic models becomes essential for process design and optimization.
In this study, we employ the Group Contribution with Association Equation of State
(GCA-EOS) to comprehensively model the phase equilibria of CO2 + carboxylic acid homo-
logue series. The GCA-EOS has been successfully applied to represent the global phase be-
havior of carbon dioxide mixtures with alkanes and alcohols. In this work, we extend this
model to represent vapor−liquid, liquid−liquid, and vapor−liquid−liquid equilibria of CO2
mixtures with short and long-chain carboxylic acids, as well as dicarboxylic acids.
Through a meticulous parametrization strategy coupled with a robust thermodynamic
model, we develop a tool for assessing the multiphase behavior of CO2 + carboxylic acids.
The model is validated against an experimental database covering C2−C20 carboxylic acids,
temperatures from 308 to 473 K, and pressures up to 400 bar. Remarkably, utilizing a single
set of parameters, the model successfully predicts phase equilibria of binary mixtures not
included in the parametrization procedure.

Keywords: Carbon Dioxide, Carboxylic Acids, Phase Equilibria, Supercritical fluid extrac-
tion.

194
 Techniques to solubilize and adsorb acid dyes in alpaca (vicugna pacos)
fiber using supercritical fluid
Beatriz G. Orcón Basilioa*, Delia R. Verástegui Cholana, María E. Tito Silvaa, Fredy V.
Huayta Socantaypea, Joel L. Sedano Malpartida a, Roberto Chaveza, Sandra M. Figueroa F.a,
S. Mark Trujillo N.a, Elizabeth V. Carranza L.a
a
Universidad Nacional de Ingeniería, Av. Túpac Amaru 210 – Rímac, Lima, Perú
* corresponding author email address: borcon@uni.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
In this research, it was qualitatively analyzed a technique to solubilize and adsorb acid dyes
using supercritical fluid. The first test was diffusion and dilution of the acid dye in supercritical
fluid, for which the acid dyes Nylosan red N-2RBL from Achroma, Erionyl red A-3G, which
have an ionic nature, and Disperse Blue SR 300% which has a nonpolar nature. Each dye, 0.1
gram, was separately wrapped with a polyester fabric. No co-solvent was dosed and only
supercritical CO2 was used. In this test, the basic model Helix SFE System supercritical fluid
equipment was used. (Applied Separations brand). The results showed that the disperse dye
Disperso Blue SR 300% exhibited slight diffusion in the alpaca fiber, however it faded as the
days passed, while the Nylosan red N-2RBL from Achroma and the Erionyl red A-3G did not

195
solubilize. The second experimental test consisted in solubilizing and adsorbing the acid dye
in a homogeneous way in the alpaca fiber. In this sense, only Nylosan Red N-2RBL and
Nylosan Yellow N-3RL were used. Here, 4 possible techniques were used to achieve good
solubility and adsorption and fastness to washing was used as a fixing indicator, which was
done with a China Base Infrared laboratory dyeing machine, model HTN-24P. The 4
techniques are described below:
Technique No. 1: The alpaca fiber sample is not pre-treated. The dye is added wrapped in a
polyester cloth (the dye does not have direct contact with the alpaca fiber) it is entered into the
supercritical fluid machine, water is dosed as co-solvent and it is operated under supercritical
conditions.
Technique No. 2: The alpaca fiber sample is not pre-treated. The dye is added in small doses
(the dye has direct contact with the alpaca fiber), it is entered into the Supercritical Fluid
machine and acidulated water is added as co-solvent and it is operated under supercritical
conditions.
Technique No. 3: A pre-treatment is applied to the sample of alpaca fiber, which consists of
soaking the alpaca fiber in a bath of acidulated water and then passing it through a roller which
allows obtaining moistened alpaca fiber. Powdered dye is added to this moistened alpaca fiber.
Then, the alpaca fiber, moistened and powdered with dye, is entered into the Supercritical Fluid
machine and is operated under supercritical conditions without dosing co-solvent.
Technique No. 4: A pre-treatment is applied to the sample of alpaca fiber, which consists of
soaking the alpaca fiber in an acidulated water bath. To this previously acidulated water bath,
the dye is added, then it is passes through a roller managing to obtain moistened and colored
alpaca fiber. This moistened alpaca fiber is entered into the Supercritical Fluid machine and
worked under supercritical conditions with a co-solvent of acidulated water.
The results show that acid dyes cannot be solubilized in supercritical CO2 and need a co-
solvent. With technique 1 and 2 there is a slight solubility, but it is not adsorbed on the fiber,
therefore a solidity measurement was not made. Technique 3 gives better solubility and adsorbs
to fiber, has a solidity of 1, but does not look good. Technique No. 4 was the best, with this
technique good solubility and adsorption was achieved, it looks good because the color is
uniform in the fiber and it presented a 3.5 degree wash fastness, which is a good indicator to
develop supercritical fluid dyeing processes for alpaca fibers.

Keywords: Solubility, Adsorption, Supercritical CO2, acid dyes, alpaca fiber, Dyeing with
supercritical fluid.

196
 Operation parameters of dyeing alpaca fiber with supercritical
fluid
Beatriz G. Orcón Basilio*, Delia R. Verástegui Cholan, María E. Tito Silva, Fredy V.
Huayta Socantaype, Joel L. Sedano Malpartida, Sandra M. Figueroa Flores, S. Mark
Trujillo Navarro, Elizabeth V. Carranza López

Universidad Nacional de Ingeniería, Av. Túpac Amaru 210 – Rímac, Lima.

*corresponding author email address: borcon@uni.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
Supercritical carbon dioxide (SC-CO2) was used on alpaca fiber as an alternative dyeing
process in order to quantitatively analyze the variation of the wash fastness of the alpaca fiber
dyeing as a function of the operating parameters and the identification of the parameters that
significantly influence the supercritical fluid dyeing process. For this, a pre-treatment is carried
out, which consists of immersing the alpaca fiber in an acidulated solution in which the Nylosan
Yellow N-3RL dye was dissolved, then being squeezed by a roller to have a moistened and
colored fiber, subsequently it is placed in the 50 ml vessel of the supercritical fluid equipment.
The influence of the dye concentration in the acidified water on the fastness to washing in the
dyeing process with supercritical fluid was evaluated. It was evaluated in the following
concentrations: At 4,228 x 10-4 g/ml and 13,032 x 10-4 g/ml of Nyolsan Yellow N-3RL dye and
the influence of pressure, temperature and time in the dyeing process was also evaluated. in the
basic model Helix SFE System supercritical fluid equipment under a statistical experimental
design of factorial type of 3 variables (operation parameters) at 2 levels: Pressure (100 and 350

197
bar), temperature (60 and 90 °C) and time (30 and 120 minutes) of which the fastness to
washing is evaluated under the ISO 105-C06-B1M standard. The results were that the fastness
is 4.5 when the concentration in the acidulated water of Nylosan Yellow N-3RL dye is 13.032
x 10-4 g/ml and from the statistical result, temperature is the operating parameter that
significantly influences the supercritical dyeing process as it obtains a p-value of 0.035,
compared to pressure (p-value 0.156) and time (p-value 0.261), obtaining a wash fastness of 4
at a pressure of 100 bar, temperature of 90 °C and time of 30 min. From this it can be said that
at higher concentration of dissolved dye in acidulated water during pretreatment, greater
fastness is obtained and that wash fastness depends on varying temperature in the supercritical
dyeing process of alpaca fiber.

Keywords: Alpaca fiber, supercritical fluid dyeing, Nylosan Yellow N-3RL dye, Wash
fastness.

198
 An integrated high-pressure and supercritical CO2-based
process for sterilization of alginate-gelatin aerogels

Cristiana S. A. Bentoa, Susana Alaricob,c, Nuno Empadinhasb,c,

Hermínio C. de Sousaa, Mara E. M. Bragaa*
a
University of Coimbra, Chemical Process Engineering and Forest Products Research Centre
(CIEPQPF), Department of Chemical Engineering, 3030-790 Coimbra, Portugal
b
University of Coimbra, Center for Neuroscience and Cell Biology (CNC) and Center for Innovative
Biomedicine and Biotechnology (CIBB), Rua Larga, Faculty of Medicine, Polo I, 1st floor, 3004-504
Coimbra, Portugal.
c
University of Coimbra, Institute for Interdisciplinary Research (IIIUC), Casa Costa Alemão - Pólo
II, 3030-789 Coimbra, Portugal
*marabraga@eq.uc.pt

GRAPHICAL ABSTRACT

ABSTRACT
Due to their numerous advantageous functional properties, biopolymers have been widely
proposed to produce several composite biomaterials and/or scaffolds for distinct biomedical
and tissue engineering applications [1]. Still, their application in these fields has been limited
due to the physicochemical degradation that may occur during most of the currently available
and conventional processing/sterilization methods. For instance, traditional sterilization
procedures such as steam heat, γ-radiation, ethylene oxide and others may compromise the
physicochemical, morphological and mechanical properties of many biopolymers [2].
The effectiveness of supercritical carbon dioxide (scCO2) inactivating microorganisms has
already been widely reported [3] and the method has been proposed as an alternative
disinfection, or even sterilization technique for chemical-, moisture- and/or heat-sensitive
materials such as biopolymer-based materials. Although it has not yet been approved as a
sterilization method by European entities, such as the European Pharmacopeia, it clearly offers
real advantages when compared to conventional methods.

199
Furthermore, scCO2 can be used as a drying method to produce aerogels [4]. Due to their
hydrophilic natures, the scCO2 drying of biopolymers typically requires an additional
processing step, namely a water-alcohol (typically ethanol) solvent exchange to form an
alcogel. This process usually requires long processing times. Yet, it is possible to employ a
high-pressure solvent exchange (HPSE) method to form these alcogels, therefore accelerating
mass transfer and reducing the duration of this step [5].
By integrating the aforementioned high-pressure solvent exchange and scCO2-based
methodologies (HPSE, scCO2 drying and sterilization), it will be possible to prepare and
sterilize aerogels in shorter processing times and at lower costs.
In this study, HPSE, and scCO2 drying and scCO2 sterilization were integrated to prepare
alginate-gelatin sterilized aerogels. The physicochemical and mechanical properties of the
produced aerogels were evaluated. The efficiency of the scCO2 sterilization was also assessed,
by contaminating the samples with Bacillus atrophaeus spores. Two scCO2 sterilization
conditions were tested (different pressure cycles and depressurization rates) and the samples
were processed inside sterilization pouches. Results showed that the sterilization procedure did
not compromise the chemical and thermal properties of the aerogels (as determined by FTIR
and DSC analysis). The swelling properties of the aerogels were also unaffected by this
sterilization procedure. Contrarily, the mechanical properties and BET surface area were
affected by this sterilization procedure (BET surface area decreased, and Young’s Modulus
increased), due to structural changes caused by the rapid pressurization/depressurization rates
applied. Regarding sterilization efficiency, the results showed a reduction in contamination
throughout the process, achieving a SAL-10-4.
In conclusion, the integrated process employed in this work successfully produced sterile and
ready-to-use aerogels that can be further tested for biomedical and tissue engineering
applications.

Keywords: process integration; high pressure solvent exchange (HPSE); scCO2 drying &
sterilization, alginate-gelatin aerogels

REFERENCES
[1] R. Song, M. Murphy, C. Li, K. Ting, C. Soo, Z. Zheng, Current development of biodegradable
polymeric materials for biomedical applications, Drug Des. Devel. Ther. 12 (2018) 3117–3145.
https://doi.org/10.2147/DDDT.S165440.
[2] C.S.A. Bento, M.C. Gaspar, P. Coimbra, H.C. De Sousa, M.E.M. Braga, A review of
conventional and emerging technologies for hydrogels sterilization, Int. J. Pharm. 634 (2023)
122671. https://doi.org/10.1016/j.ijpharm.2023.122671.
[3] N. Ribeiro, G.C. Soares, V. Santos-Rosales, A. Concheiro, C. Alvarez-Lorenzo, C.A. García-
González, A.L. Oliveira, A new era for sterilization based on supercritical CO2 technology, J.
Biomed. Mater. Res. - Part B Appl. Biomater. 108 (2020) 399–428.
https://doi.org/10.1002/jbm.b.34398.
[4] I. Smirnova, P. Gurikov, Aerogel production: Current status, research directions, and future
opportunities, J. Supercrit. Fluids. 134 (2018) 228–233.
https://doi.org/10.1016/j.supflu.2017.12.037.
[5] A. Lebedev, E. Suslova, A. Troyankin, D. Lovskaya, Investigation of aerogel production
processes: Solvent exchange under high pressure combined with supercritical drying in one
apparatus, Gels. 7 (2021) 1–16. https://doi.org/10.3390/gels7010004.

200
 Development of Simulator: Supercritical Extraction Process
Otávio A. F. Oliveira, Rubem M. F. Vargas, Eduardo Cassel*

Unit Operation Lab, School of Technology, PUCRS, Ipiranga, 6681, Porto Alegre, Brasil
*cassel@pucrs.br

GRAPHICAL ABSTRACT

ABSTRACT
Supercritical extraction is a process that has grown in terms of industrial applications,
especially in sectors involving food production, but technical and economic feasibility studies
are still incipient.Therefore, the development of a calculation tool that allows economic
analysis and process scale up based on mass and energy balances, mass transfer models, fluid
properties, experimental extraction results and data introduction economics defines the purpose
of this work. In this development, the case study is the extraction of caffeine from yerba mate,
using carbon dioxide as a solvent and ethanol as a co-solvent, based on data from a pilot-scale
equipment. The following information used for the development of the simulation tool is
highlighted: the properties of carbon dioxide and ethanol in the extraction conditions, the
energy balances of the unit operations that constitute the process: pumps, heat exchangers, and
valves, and the mass balances of the chains that interconnect the equipment. The input variables
defined in the developed simulator are the following: extraction time, mass of raw material,
mass flow rate of solvent, mass flow rate of co-solvent, pressure, temperature and process
operation time. The scale-up process was carried out using the equality between the ratio of

201
CO2 flows in pilot and industrial scales and the ratio of raw material masses in both scales.
Solvent properties such as vapor pressure, density and viscosity were obtained through the
Aspen Hysys process simulator (V 8.0), using the EOS Peng-Robinson and heat exchangers
were designed using EDR software (v8.0). The simulator was built on an Excel® spreadsheet.
From the introduction of the previously mentioned input variables, the properties of the raw
material, the extract and the solvents and the introduction of mass and energy balance equations
for processes in steady state, it was possible to calculate, in an automated way, the energies and
masses involved in each process equipment, as well as the associated monetary costs.

Keywords: scale-up, yerba-mate, economic analysys, mass balance, energy balance.

202
 Bioactive Compounds from Yerba-Mate: Supercritical
Decaffeination and Polyphenol Extraction
Bruna Sgarioni, Luane C. Angeli, Rubem M. F. Vargas, Eduardo Cassel*

Unit Operations Lab, School of Technology, PUCRS, Ipiranga, 6681, Porto Alegre, Brazil
*cassel@pucrs.br

GRAPHICAL ABSTRACT

ABSTRACT
Numerous studies have focused on obtaining bioactive compounds from plant-based raw
materials. These researches explore the use of various solvents, such as water, supercritical
CO2, and organic solvents, and discuss different extraction methods, evaluating how the choice
of technique influences in extraction of the desired compound. Yerba-mate has been
extensively studied as a plant source due to its observed antioxidant capacity in extracts, which
is attributed to the presence of phenolic compounds. The composition of Ilex paraguariensis
extract is influenced by both cultivation and plant processing, as well as the extraction method
employed. The main bioactive substances obtained include polyphenols, saponins, and
methylxanthines. According to literature reports, yerba mate extract contains up to 58
polyphenols and 3 methylxanthines. To maximize the phenolic composition of the extract, the
process of decaffeination using supercritical fluid extraction is employed, as this method
selectively removes alkaloids while preserving the phenolic composition of the plant.
Therefore, the aim of this study was to extract polyphenols using water infusion from
decaffeinated yerba mate, and to encapsulate them in gelatin using a spray dryer. The
evaluation of polyphenol release of the capsules was carried out in a simulated gastric fluid.
The decaffeination process of the yerba-mate leaves was carried out through supercritical fluid
extractions, optimized using a Box-Behnken experimental design, varying the CO2 flow rate,
cosolvent flow rate, and extraction time. This design was based on previous studies conducted
by our research group, using in natura vegetal matrix. In this work, the Uruguayan variety of
yerba mate was used, with the extraction pressure, temperature, and mass kept constant at 300
bar, 60 °C, and 140 g, respectively. The response generated was the caffeine concentration in
the vegetal matrix, which was determined by characterizing the aqueous extract by HPLC
analysis. For polyphenol extraction, an orthogonal factorial design was conducted, with the
total polyphenol content in gallic acid equivalents (GAE) as the response. In this design, the

203
variables used were the extraction time and temperature. The extractions were performed using
an infusion system of decaffeinated plant material in an aqueous environment with reflux. In
addition, the antioxidant capacity of the extracts was also evaluated. With the polyphenol-rich
extract, encapsulation in gelatin was proposed. The results obtained for optimized conditions
of the decaffeination process were as follows: CO2 flow rate of 950 g/h, a cosolvent flow rate
of 1.643 mL/min, and an extraction time of 4.7 h. These results were consistent with those
obtained in the design to yerba-mate in natura, and it is worth noting that the caffeine
concentration in yerba mate after the decaffeination process was 0.01% (g caffeine/g yerba
mate). Based on this result, it can be stated that a decaffeinated plant was obtained according
to ANVISA guidelines (caffeine content < 0.1% g caffeine/g plant). Furthermore, the
polyphenol extraction, using water infusion process, was optimized under the following
conditions: 70 °C and 30 min. This result is under expectations since the ideal temperature for
consuming cultural yerba mate-based beverages (chimarrão, mate) is observed as the optimized
conditions. Additionally, it was observed that the extraction temperature is the most important
variable in the process, as the surface generated by the experimental data did not show
variations in the response for different extraction times at the optimized temperature. Beyond
that, in the optimized condition polyphenol-rich extract concentration observed was 1.6% (mg
polyphenol in GAE/mg extract). The conducted experiments emphasize the importance of
obtaining bioactive compounds from yerba mate, with an emphasis on polyphenols. The use of
suitable solvents and extraction methods influences the composition of the extract, with
supercritical fluid decaffeination being an efficient process to preserve the phenolic
composition of the plant. This study provides important knowledge, mainly, for the food,
cosmetics and supplement industries, leading off for the production of more effective products
with beneficial properties for human health.

Keywords: caffeine; Ilex paraguariensis; water infusion; polyphenols; spray dryer.

204
 Effect of Casearia sylvestris supercritical extracts on levels of
serum cholesterol, triglycerides and glucose
Patrícia Benellia,*, Rozangela C. Pedrosab, Sandra R. Salvador Ferreirab
a
ICTA/UFRG: Institute of Food Science and Technology, Federal University of Rio Grande do Sul -
UFRGS, Porto Alegre/RS, Brazil.
b
EQA/UFSC: Chemical and Food Engineering Department, Federal University of Santa Catarina -
UFSC, Florianopolis/SC, Brazil.
*Corresponding author: patricia.benelli@ufrgs.br

GRAPHICAL ABSTRACT

ABSTRACT
Natural antioxidant compounds have been extensively studied and isolated from different
plant materials. These compounds include flavonoids, phenolic acids, terpenes, tocopherols
and phospholipids. Casearia sylvestris is a native medicinal plant in Brazil, Peru, Argentina,
Uruguay and Bolivia and its leaves are popularly used in folk medicine as antiseptic, topical
anaesthetic, antitumor and antiulcer agents, and to heal skin wound diseases. C. sylvestris
extracts presents substances of great interest, such as coumarins, flavonoids and diterpenes,
especially clerodane diterpenes as casearins and casearvestrins, which are bioactive
compounds with excellent cytotoxic and antitumor potential. Some substances present
hypocholesterolemic, hypotriglyceridemic, hypoglycemic and anti-obesity effects.
Considering this, the aim of the present study was to obtain and compare the application of C.
sylvestris extracts recovered by supercritical fluid extraction (SFE) with CO2 (300 bar, 50 °C
- ESC300/50) and CO2 with co-solvent (300 bar, 50 °C and 5% of ethyl acetate (w/w) -
ESC300/50EA) in terms of reduction of blood cholesterol, triglycerides and glucose levels,
and reduction of intestinal absorption of triglycerides. These SFE extracts were selected due
to their best anti-obesity results in previous studies. The effect of the extracts was evaluated

205
by incorporating them into a high-fat diet to Mus musculus Balb/c mice (in vivo assays) using
100 mg/kg/day of C. sylvestris extracts during 30 days, monitoring the animals’ weight and
the final total serum cholesterol, triglycerides and glucose levels. The animals were divided
in four groups: ESC300/50 (high-fat diet combined with C. sylvestris extract); ESC300/50EA
(high-fat diet combined with C. sylvestris extract); CNDN (negative control treated with
normal diet); CNDH (negative control treated with high-fat diet). All groups received water
ad libitum. The inhibition of intestinal lipid absorption experiment by supercritical extracts
(ESC300/50 and ESC300/50EA) was also conducted using Mus musculus Balb/c mice treated
with olive oil and compared to Orlistat (tetrahydrolipstatin) on triglyceride levels. All animals
were maintained and handled in accordance with the legal requirements and
recommendations applicable to the species, in accordance with the Guide for the Care and
Use of Laboratory Animals (NIH, 2011), and with permission from the Commission on
Ethics in the Use of Animals (CEUA) of UFSC (research protocol number PP00744). For
cholesterol levels, both extracts showed values significantly lower than the cholesterol level
found for the negative control treated with hypercaloric diet (CNDH), but higher than the
negative control treated with normal diet (CNDN). For triglycerides levels, lower levels were
obtained than those obtained for the negative control treated with a hypercaloric diet
(CNDH), and the group treated with ESC300/50EA extract was the one that more reduced
triglycerides levels, being statistically equal (p < 0.05) to the negative control (CNDN). It is
important to note that elevated blood levels for triglycerides have a direct correlation with the
deposition of these lipids in adipose tissue and, consequently, with obesity. The reduction in
circulating levels of triglycerides in animals treated with ESC300/50EA extract corroborates
with previous studies where the lowest weight gain occurred in this same group of animals
and suggests a preventive effect against obesity for this extract. Glucose levels were
significantly below to CNDH group and these results also showed that the extracts
maintained the glycemic level equal (p < 0.05) to CNDN and ESC300/50EA extract was
responsible for reductions of 26.1% and 9.75% in relation to the CNDH and CNDN groups,
respectively. The inhibition of intestinal lipid absorption experiment showed that C. sylvestris
extracts promoted significant reduction in serum triglyceride levels after, suggesting a
hypolipidemic effect of the extracts. These results indicate that these extracts have an
inhibitory effect on the absorption of triglycerides in the intestinal mucosa since, even after
the administration of olive oil (rich in triglycerides), the blood level of this lipid remained
below the negative control in all periods of time evaluated. Based on the results, it can be
stated that ESC300/50EA extract from C. sylvestris showed anti-obesity activity since it
reduced triglycerides, cholesterol and glucose levels, probably by inhibiting their absorption
at intestinal level.

Keywords: supercritical extract, hypolipidemic effect, anti-obesity, hypercaloric diet,

intestinal absorption.

206
 Processing of rosehip and grapes yields for recovery of
biologically active components
Maša Knez Marevcia*, Željko Kneza,b
a
University of Maribor, Faculty of Chemistry and Chemical Engineering, SI-2000 Maribor, Slovenia,
b
University of Maribor, Faculty of medicine, SI-2000 Maribor, Slovenia
*corresponding author: masa.knez@um.si

GRAPHICAL ABSTRACT

ABSTRACT
Investigation of biologically active components of various natural materials such as fruits and
vegetables represents a great perspective due to multiple health benefits and dynamic demand
of the food and cosmetic industries. Therefore, numerous extraction methods have been
employed and modified to provide high-quality extracts that offer health benefits when used in
food or cosmetics. Conventional extraction techniques such as cold maceration (CM) and
Soxhlet extraction (SOX) are well considered for most natural materials, but recently more
sustainable methods such as ultrasonic and supercritical fluid extraction (SCE) have been
extensively researched. The use of environmentally friendly, non-hazardous solvents is
essential to obtain a safe product for the consumer while ensuring the sustainability of the
manufacturing process. Supercritical extraction with CO2 is used as an extraction technique
with application in food processing for aroma and flavor recovery, extraction and fractionation
of edible oils and removal of contaminants. The cost of SCF is becoming competitive for
largescale operations. One of the most sought-after components of natural extracts are
polyphenols, phytochemicals known for their numerous beneficial bioactivities. They behave
as antioxidants which means that they prevent diseases caused by oxidative stress, exhibit anti-
aging activity, and have cardioprotective, anti-cancer, anti-inflammatory, and antimicrobial
activity. Polyphenols comprise a significant portion of plant antioxidants but aren't the only of
the major groups. The quality of extracts has been examined as a function of the type of
material, extraction technique and solvent in several studies. Moreover, the qualitative and

207
quantitative profiles of extracts vary widely even within the same species depending on
extraction conditions, plant location, or environmental factors. Rosehip fruit as well as white
and red grapes are known of their high levels of natural polyphenols, well-documented
antioxidant activity that are traditionally used to treat a variety of ailments. Besides, these
materials are widely available in the phytogeographical region of Central and South – Eastern
Europe. Moreover, the byproducts of wine production, which is an important agricultural
activity in the region, can be used for valorization of functional ingredients or bioactive
phytochemicals, that can be devoted to the generation of pharmaceutical, food, and cosmetic
ingredients. In this study the compositions of extracts, prepared by three different extraction
methods have been examined. Cold maceration and Soxhlet extraction were performed using
EtOH and MeOH as solvents. Supercritical extraction with CO2 was performed using EtOH
as co-solvent. Processing of rosehip and grapes yields a significant portion of waste such as
skins, seeds, and stalks, which also retain large amounts of biologically active components.
Therefore, these primary and secondary raw materials have great potential as sources of desired
phytochemicals, improving resource recovery efficiency. As natural resources become
increasingly scarce and the climate crisis becomes a forefront issue, more and more efforts are
made to intensify nutrient extraction from by-products and incorporation of these compound
into stabile formulations. There is still an issue how to transfer these concepts efficiently on a
large scale. Sustainable processing, combined with a sustainable economy, requires innovative
solutions that suggest and evaluate feasible solutions from a technological point of view.
Extractions from red and white grapes skin and the rosehip (Rosa canina) have been performed
to acquire high phenolic content extracts. Extraction with supercritical fluids (SFE) and
conventional extraction methods have been carried out and the influence of extraction method
and operating parameters (time, pressure, temperature, and solvent mixture) have been
evaluated. Total phenolic content, proanthocyanins, and the phenolic constituent profile has
been examined. Additionally, antioxidant capacity has been measured and evaluated as a
function of the content of active compounds. Their contents significantly varied among the
different types of materials. The highest content of total phenolics was determined in the extract
from the white grape skin, macerated with MeOH (26.7 mg GA/g extract), and similar, 25.6
mg GA/g extract in the MeOH extract attained by Soxhlet. Ellagic acid (0.650 mg/100 g
extract), catechin (0.164 mg/100 g extract), gallic acid (0.133 mg/100 g extract) as well as
caffeic acid (0.038 mg/100 g extract) are the main compounds present in the rosehip extracts
attained by maceration using MeOH as solvent. On the other hand, supercritical fluids as
processing media allow selective extraction of very pure, high-value products from materials
which otherwise would be considered byproducts or waste. Such processing concepts promote
biorefinery concept which comprises reuse of residues from food industry.
The results show that the materials that represent wide amounts of wastes in food and beverage
industry, contain important types of phytochemical constituents with antioxidant potential.
Extracted fractions possess the antioxidant potential and might be helpful in preventing or
slowing the progress of various oxidative stresses. By studying the process kinetics at varying
operational conditions, optimized process parameters are determined experimentally, at which
the target bioactive substances are more completely extracted, and at which the extracts show
the highest antioxidant capacity. This paper supplies useful information for improving the
extraction process to produce enriched extracts from rosehips and grape skin with a higher
content of antioxidant substances beneficial to human health.

Keywords: High pressure, plant extracts, biorefinery concept, polyphenols, phytochemicals.

208
 Valorization of grape marc (Vitis labrusca) using supercritical
extraction: Lab-scale DOE and pilot-scale validation
Camilo Pardo-Castaño a,b *, Edwin Sanchez a and William Vallejo c

a
Didacontrol SAS, Cl. 32 #1-22, Cali, Colombia.
b
Universidad del Valle, Cl. 13 100 -00, Cali, Colombia
c
SENA, Cl. 52 No. 2-Bis-15, Cali, Colombia

*corresponding author: camilo.pardo@correounivalle.edu.co

GRAPHICAL ABSTRACT

ABSTRACT

The winemaking and grape juice industry generates a considerable amount of grape marc,
exceeding 20 million tons per year. Some of it is typically utilized as animal feed and fertilizer,
while the remainder is discarded as a residue, resulting in environmental damage such as gas
and odor emissions, soil degradation, and water pollution. In recent years, grape marc has
emerged as a promising alternative source for obtaining high-value materials due to its
bioactive content, which exhibits potent antioxidant and antimicrobial properties.
Consequently, it is crucial to develop environmentally friendly methods for its valorization,
contributing to one of the world’s most important goals: sustainability.
In this study, the extraction of bioactive compounds from grape marc (Vitis Labrusca) was
conducted at a laboratory scale using a 350 mL extractor. The extraction was carried out using
Supercritical Fluid Extraction (SFE) with and without ethanol as a cosolvent. A 24-1 fractional
factorial design with central replicates was developed to determine the impact of temperature
(30 to 60 °C), pressure (138 to 276 bar), cosolvent concentration (0 to 10 wt%), and average
particle size of the dried grape marc (116 to 601 μm) on the antioxidant capacity, total phenolic

209
content, and extract yield. The aim was to identify the optimal operating conditions for the SFE
process within the specified experimental range.
The analysis of variance (ANOVA) indicated that all the variables have a statistically
significant effect on the measured response variables, with a significance level (α) greater than
4%. The best experimental results were achieved with a yield of 9.2 wt%, antioxidant activity
of 238.4 μmol trolox eq./g, and phenolic content of 64.0 mg GA eq./g. These results were
obtained at a temperature of 60°C, pressure of 138 bar, 10 wt% ethanol concentration, and an
average particle size of 116 μm, with an extraction time of 2.5 hours. Additionally, using only
CO2 as the solvent at 30°C, a pressure of 276 bar, and an average particle size of 601 μm, a
yield of 2.5 wt%, antioxidant activity of 365.3 μmol trolox eq./g, and phenolic content of 69.3
mg GA eq./g were obtained in the same extraction time of 2.5 hours.
A supercritical extraction pilot plant was designed and constructed with a specific solvent-to-
feed (S/F) ratio of 0.4 min-1. The pilot plant has a capacity of 20 L and consists of two 10 L
extractors with a height-to-diameter (H/D) ratio of 5, along with two 3 L separators. It offers
the flexibility to operate in a semiautomatic mode, controlled by an intuitive control panel. The
extractors and separators can be operated in series or parallel configurations. The pilot plant is
capable of handling pressures up to 345 bar and temperatures up to 100 °C.
To ensure safety and performance, several tests were conducted to validate the operation of the
pilot plant. These tests demonstrated that the yields achieved at the laboratory scale can be
reproduced at the pilot scale, and in some cases, even improved. For example, an extraction
yield of 2.9 wt% was obtained in just 2 hours at a temperature of 30 °C, pressure of 276 bar,
and an average particle size of 770 μm. These findings indicate that the pilot plant is capable
of successfully scaling up the extraction process. The pilot plant's operation in a semiautomatic
mode, along with its intuitive control panel, further enhances its usability and ease of operation.
The presented results offer promising prospects for obtaining high-value products from grape
marc, especially in regions like Colombia where the Vitis labrusca species is prevalent. This
development holds the potential to address environmental concerns while also enhancing
profitability within the winemaking and grape juice industry. By effectively utilizing grape
marc, which is currently considered a byproduct, this approach can contribute to both
environmental sustainability and economic gains in the sector.

Keywords: grape marc, supercritical extraction, carbon dioxide, ethanol, antioxidants.

210
 ENCAPSULATION OF NIACIN IN PVP-K30 BY GAS
ANTISOLVENT TECHNIQUE
Marcel Silveiraa, Patricia V. Oliveiraa, Evertan A. Rebelattoa, Pedro H. H. Araújoa, Carlos E.
M. Camposb, J. Vladimir Oliveiraa*
a
Department of Chemical and Food Engineering, Universidade Federal de Santa Catarina-
UFSC, Florianópolis, SC, Brazil
b
Department of Physics and X-Ray Diffraction Laboratory, Universidade Federal de Santa
Catarina-UFSC, SC, Brazil
*corresponding author email address: jose.vladimir@ufsc.br

GRAPHICAL ABSTRACT

ABSTRACT
Nicotinic acid, also known as vitamin B3 or vitamin PP or niacin, is a well-known compound
in the medical literature for treating LDL (low-density lipoprotein) cholesterol reduction. It
also helps in the proper functioning of enzymes and the treatment of pellagra. This compound
can be released into the body by immediate or prolonged release. Niacin can be found in plants
and animals. It can be used as a food additive in industry to enrich wheat and corn flour. It also
has applications in the area of veterinary medicine for ruminant animals. In dairy cows, niacin
is the subject of studies to avoid production losses due to heat stress. Nicotinic acid may have
side effects in patients such as skin flushing. The encapsulation of nicotinic acid can help
reduce these effects through the mechanism of controlled release of this drug. In ruminant
animals, a rapid or controlled release may be an interesting target for studies due to the greater
complexity of the digestive tract of these animals. Carbon dioxide is the supercritical fluid most
commonly mentioned in the literature in drug processing. Precipitation of the compound of

211
interest by supercritical fluid techniques results in products with a trace level of solvent,
generally mild temperatures are used, and it is very versatile for use with polar and nonpolar
drugs. This study, aimed to form solid dispersions of niacin in PVP-K30 by the gas antisolvent
technique (GAS). A Box-Behnken factorial design with 15 conditions and three central points
was performed to study the effect of the independent variables: pressure (12, 14 and 16 MPa);
temperature (35, 40 and 45 oC), and encapsulation ratio [encapsulant/encapsulated] (1:1, 1.5:1,
and 2:1) on particle size, process yield and encapsulation efficiency. Each condition was
analyzed via FTIR, DSC and XRD. Scanning Electron Microscope analyses are underway
toward defining the most appropriate encapsulation processing conditions regarding the
morphological nature of particles produced and their size. GC-MS analyzes will be performed
to evaluate the dissolution rate of each proposed in factorial design. XRD results showed that
the final product has a degree of crystallinity between (78-84) %, approximately, and important
structural and microstructural changes of the nicotinic acid as compared to its pure form. DSC
analyses showed that in some formulations there were shifts in the melting point, for example:
variation from 237oC to close to 217oC (onset) and also to close to 200oC indicating that
specific structural and microstructural changes of niacin can favor its solubility in the
formulation. Spectroscopic analyzes indicate new intermolecular forces occurring in the
formed solid phase. A broad peak at 3385.17 cm-1 was associated to hydrogen bond formation.
The peak at 1699.34 cm-1 associated with the asymmetric stretching of nacina (COO-) bond is
not present in the encapsulations. A peak shift from 1651.59 to 1647.04 cm-1 also appears,
which may indicate a weakening of the PVP amide C=O bond. The preliminary results of the
FTIR, DSC, and XRD studies already show the success of the new solid solution between
niacin and PVP prepared by the GAS technique and confirm what has been demonstrated in
the literature for other drugs and encapsulants.

Keywords: Supercritical fluid, niacin, encapsulation, Box-Behnken factorial design

212
 Calculation of critical endpoints and phase diagrams of highly
asymmetrical binary mixtures
Francisco A. Sáncheza,b, Martín Cismondic,*
a
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca, (8000), Argentina
b
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253, Bahía
Blanca, Argentina
c
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA),
Universidad Nacional de Córdoba (UNC) – CONICET, Córdoba, Argentina
*corresponding author email address: martin.cismondi@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
The determination of monovariant lines like critical and three-phase equilibrium curves in
phase diagrams of binary mixtures plays a fundamental role in analyzing potential process
conditions and assessing the qualitative and quantitative capabilities of thermodynamic
models. In this regard, the automatic generation of global phase equilibrium diagrams
(GPED) has become an invaluable analytical tool, enabling intelligent solutions to highly
complex and nonlinear mathematical problems. In particular, it was mainly the software
GPEC (Global Phase Equilibrium Calculations) and the publications describing its
computation strategies and methods that made it possible for a large international community
of researchers to use GPED’s as frequent tools in the application of equations of state to
different problems.

213
However, the original GPEC algorithm face challenges in resolving highly asymmetric
mixtures, such as gases (CH4, C2H6, CO2, N2) in combination with heavy paraffins, vegetable
oils, or even polymers, as well as electrolytic aqueous systems. These challenges manifest in
convergence problems and floating point overflow. More specifically, the UCEP or “k point”
located quite close to the pure gas critical point (phase behavior of type III or IV) frequently
fails to be properly found and converged in this type of systems due to the low concentration
of the solute in the critical phase.
In this study, we present an innovative calculation methodology to address these challenges
in highly asymmetric binary systems. Our approach builds upon the extension of existing
algorithms, aiming to achieve an automatic generation of GPED that does not require manual
intervention. The approach is based on the formulation of other phase equilibrium problems
dealing with nearly pure phases. As a case study, we apply our methodology to analyze phase
equilibrium transitions in industrially relevant systems, such as CO2 with heavy alkanes.
Keywords: asymmetrical binary mixtures, critical lines, critical end points, LLV, EOS

214
 Coumarins Supercritical Extraction from Citrus Peel
Juan J. Mora, Henrique M. Tavares, Eduardo Cassel, Rubem M.F. Vargas*

Unit Operations Lab, School of Technology, PUCRS, Ipiranga, 6681, Porto Alegre, Brazil
*rvargas@pucrs.br

GRAPHICAL ABSTRACT

ABSTRACT
Brazil is the second biggest producer of citric fruits in the world and the biggest orange and
orange juice producer. The production of juices and other fruit products generates a large
amount of waste consisting mainly of fruit peels. These peels are a source of essential oil, but
there are also reports about the presence of molecules of interest in the development of other
natural products such as coumarins. Coumarins are a class of secondary metabolites, that are
mainly used as a fragrance enhancer. Despite its applications in aromas industry, depending on
the compound pattern, the coumarins also play a significant role in cancer prevention and
treatment. The aim of this study was to find the optimal conditions of pressure, temperature
and granulometry for the supercritical fluid extraction using Citrus limonia and Citrus sinensis
peel. Also, the study of the extraction kinetics was conducted using mass transfer based
mathematical models, as well as the characterization of the extract, aiming mostly the
identification of coumarins. The extraction experiments were carried out in a pilot unit, based
on a Box-Behnken design. The citrus peels were dried out at low temperature (35 °C), milled
and a granulometric separation was performed, obtaining particles with 0.5, 1.0 and 1.5 mm of
diameter. Prior to the supercritical extraction, extractions were conducted with temperature and
pressure conditions close to the CO2 critical point (80 bar and 40 °C). For the Box-Behnken
extractions, the conditions were 100, 150 and 200 bar of pressure and temperature conditions
were 40, 50 and 60 °C. The optimization objective was to maximize the extraction yield. An
extraction curve was plotted for the optimal condition, and two different mathematical models
(Reverchon and Crank) were fitted to the experimental data to obtain mass transfer parameters.
The analysis of the extract compounds was carried out by high performance liquid

215
chromatography (HPLC), using a methodology for coumarins identification. Extracts obtained
in conditions close to the critical CO2 point, were identified by Gas chromatography-mass
spectrometry (GC-MS). Chlorophylls and carotenoids were also identified using UV
spectroscopy. The chlorophyll contents for the investigated species were found to be quite low,
while contents of total carotenoids in the order of 10-6 (w/w of feedstock) were obtained. A
qualitative test for the extracts obtained was performed by utilizing thin layer chromatography,
with the stationary phase being a mixture of chloroform and methanol (98:2). The test resulted
in the presence of coumarins, along with many other classes of compounds. The optimal
conditions obtained by the Box-Behnken were 100 bar, 40 °C and 1.5 mm for Citrus sinensis,
and 150 bar, 50 °C and 1.0 mm for Citrus limonia. Extractions under the optimal conditions
for both Citrus species were conducted, triplicates for each Citrus was performed. The average
global extraction yield obtained were 3.47% for Citrus limonia, and 5.30% for Citrus sinensis.
The curve fitting with the Reverchon mathematical model resulted in two estimated parameters,
kTM (internal mass transfer coefficient) and K (equilibrium constant). Citrus limonia
parameters found were kTM = 1.16x10-3 s-1, and K=7.65x10-2 m3.kg-1 with a R² of 0.978 and for
Citrus sinensis, kTM = 2.89x10-4 s-1, and K = 5.16x10-5 m3.kg-1 with R² of 0.975. The curve
fitting using Crank mathematical model for spherical geometry resulted in the value for
diffusivity of 4.68x10-12 m².s-1 with a R² of 0.925 for Citrus limonia and 1.10x10-11 m². s-1 with
a R² of 0.927 for Citrus sinensis, while when the rectangular geometry was considered, the
diffusivity found for Citrus limonia was 2.41x10-10 m².s-1 with a R2 of 0.911 and for Citrus
sinensis was 2.31x10-10m². s-1 with a R² of 0.922. The results obtained with the Reverchon
model were more accurate to fit the extraction curve, since its development considers more
variables present in the extraction, such as porosity and mainly transport phenomena associated
with the fluid phase, while Crank model only considers the mass transfer by diffusion from the
plant and the geometry assumed for the feedstock.

Keywords: Supercritical fluid extraction, coumarins, optimization, mathematical modeling.

216
 Optimization SFE process of bioactive compounds from
Myroxylon balsamum (Estoraque) seeds
Larry O. Chañi-Paucara,b,*, Felipe de A. Maiac, Godofredo R. Lobato-Calderona, Julian
Martínezd, Luiz H. Fasolinc
a
Food Engineering and Agroindustry Research Group (GIIAA), Universidad Nacional Autónoma
Altoandina de Tarma (UNAAT), road La Florida – Cochayoc, Huancucro, Junin, Peru.
b
LATSBIO – Laboratorio de Tecnología Supercrítica y Bioproductos, Instituto Científico Amazónico
del Perú (ICIAP), avenue Alameda 306, Madre de Dios, Peru.
c
Department of Food Engineering and Technology, School of Food Engineering, University of
Campinas (UNICAMP), CEP 13083-862 Campinas, SP, Brazil.
d
Laboratory of High Pressure in Food Engineering, Department of Food Engineering and
Technology, University of Campinas, UNICAMP, 13083-862, Campinas, SP, Brazil.

*Corresponding author: Larry.76728@gmail.com / Lchani@unaat.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
Peru balsam is a product marketed for its bioactive properties. It is obtained from the exudate
of the bark of the Myroxylon balsamum tree. It has antiseptic and anti-inflammatory properties
and is also used in aromatherapy. The seeds of this species also have a characteristic aroma
that has yet to be studied. Therefore, the present work evaluated the supercritical extraction
process with CO2 to determine operating conditions that maximize the recovery of the aromatic
extract from the Estoraque seeds (ES). The SFE process was evaluated considering the
variables mass flow (g CO2/min): 6.36 - 14.58, pressure (MPa): 10 – 40, and temperature (ºC):
40 - 70. The experimental assays were performed following the Box-Behnken design and were

217
analyzed following the response surface methodology. According to the experimental design,
fifteen treatments were performed, including three repetitions in the central point. The
optimization of the extraction process is carried out by the desirability function, considering
only the maximization of the response variable (global extraction yield). The ES was purchased
from Agroforestry J&G SAC in Madre de Dios, Peru. Extractions were performed with 5 g of
ground ES, using a 25 mL extraction vessel (internal diameter = 20 mm and height = 63.66
mm). According to the experimental design, the highest extraction yield was observed under
the conditions of 55 ºC, 40 MPa, and 14.58 g/min (18.9 g extract/100 g ES) and the lowest at
70 ºC, 10 MPa, and 10.5 g/ min (2.1 g extract/100 g ES). The optimization of the SFE process
considering no restriction of the parameters of the extraction process, to maximize the
extraction yield of aromatic ES extract, under these considerations, it is possible to obtain 19.5
g extract/100 g ES yield at 70 ºC, 36.1 MPa, and 14.5 g CO2/min. This yield was higher than
that obtained by the Soxhlet method (15 g extract/100 g ES), using petroleum ether as an
extraction solvent. The optimization of the SFE process by implementing the constant
temperature restriction of 55 and 40 ºC allows obtaining yields of 18.3 g extract/100 g ES (55
ºC, 33.3 MPa, and 14.5 g CO2/min) and 17.4 g extract/100 g ES (40 ºC, 30.3 MPa, and 14.5 g
CO2/min), respectively; these yield values are close to those obtained at 70 ºC. On the other
hand, the ES extract presents characteristics of oily texture to the touch, with a sweet woody
aroma and a yellowish coloration. Its composition has not yet been reported, but perhaps it has
some similarities with the resin of the bark. (main components reported in the literature: benzyl-
benzoate, Benzoic acid, 3,4-dimethoxycinnamaldehyde, cinnamic acid, and benzyl-
cinnamate). The results are promising for including ES as a source of bioactive, but the
composition and their bioactive properties need to be elucidated.

Keywords: Peru balsam, Myroxylon balsamum, Supercritical fluid extraction, Response surface
methodology

218
 Efficient flash computation of continuous sets of solid-liquid-
vapor equilibria directly related to laboratory data obtained
through the synthetic method
Matías J. Molinaa,b, José V. Mattosd, S. Belén Rodriguez-Reartesa,b,c , Lúcio Cardozo Filhod,
Marcelo S. Zabaloya,b,*
a
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253, Bahía
Blanca, (8000), Argentina.
b
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS – CONICET), Camino
“La Carrindanga” Km 7, Bahía Blanca, (8000), Argentina.
c
Dpto de Ingeniería Química, ETSEQ, Universitat Rovira i Virgili, Av. Països Catalans 19, (43007),
Tarragona, Spain.
d
Universidade Estadual de Maringá, Departamento de Engenharia Química, Av. Colombo, 5790 -
Bloco D-90, Maringá - PR, Zona 07, CEP 87020-900, Brasil
*mzabaloy@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Often in the laboratory the set goal is to measure solid-fluid equilibria for systems made of
CO2(1), a drug(3) and an organic solvent(2), at supercritical conditions, by using a variable
volume equilibrium cell. Frequently, it is decided to proceed as follows: a known amount of a
known-composition binary liquid solution of components 2 and 3 is fed into the cell, followed
by feeding a known amount of the antisolvent CO2(1). In this way, the global composition
becomes known. Then, at set temperature (T), by moving the cell piston, a high enough
pressure (P) is found, at which the system is a homogeneous fluid (F). Next, P is lowered up to
the appearance of an incipient solid (S) phase [fluid-solid (FS) equilibrium]. Sometimes,
already at the maximum operating pressure of the apparatus, solid-fluid coexistence (no
homogeneity) is found, and hence the lowering of the pressure leads to the detection of an

219
incipient vapor (V) phase in the presence of a solid phase and of a fluid phase, each of finite
size (solid-liquid-vapor (SLV) equilibrium). In this case, at the end of the experiment, the only
known information to be recorded is: T, the global composition, and the pressure P of SLV
equilibrium where the V phase is the only incipient one [1]. If an isothermal set of such
experiments covers a range of global amount of CO2 (same liquid solution fed), then, the
experimentalist essentially obtains a curve of SLV equilibrium pressure versus, e.g., global CO2
mole fraction, at set T and set global drug/solvent ratio. We recognize that the right type of
computation corresponding to a point of such curve is a ternary solid-liquid-vapor flash at zero
vapor-phase mole fraction and set temperature T (Duhem’s Theorem), being the predicted P
just one of the outcomes of the calculation. Notice that none of the SLV phase compositions is
experimentally known. The purpose of this work is to develop an efficient algorithm for
computing curves as the previously described, each in a single computer run. This work
precedes a, strictly speaking, modeling stage in which we would seek agreement between
experimental data and model predictions. The algorithm is illustrated in this work for the
system CO2(1)+ethanol(2)+acetaminophen(3) [1], where ethanol is the organic solvent. The
system of equations of an SLV flash at set temperature is built by imposing: {1} the satisfaction
of the fluid PVTx equation of state (EOS) for the L and V phases (translated Peng Robinson-
EOS, with quadratic mixing rules, in this work), {2} the isofugacity condition for each of the
3 components in the L and V phases, {3} the isofugacity condition for component 3 among the
V and S phases (the S phase is considered to be made of pure component 3, and the solid-state
fugacity is computed according to [2]), {4} mass conservation condition in the heterogeneous
system for each component, {5} summation of mole fractions equal to unity for the L and V
phases, {6} specification equation that sets the value of T, and, {7} specification equation that
sets the V phase mole fraction equal to zero. On the other hand, the system of equations that
makes possible to compute the global mole fractions includes: {a} the constraint 𝑧1 + 𝑧2 +
𝑧3 = 1, being 𝑧𝑖 the global mole fraction of component 𝑖, and {b} the specification 𝑧3 ⁄𝑧2 =
𝑐𝑜𝑛𝑠𝑡, which corresponds to the composition of the fed liquid solution. In this work we use the
enlarged system of equations (ESEs) resulting from adding the two last equations to the SLv
system of equations. The ESEs has a single degree of freedom that one would tend to spend by
setting the value of one of the 𝑧𝑖 s. However, since we resort, for solving the ESEs, to a
numerical continuation method (NCM), due to its efficiency, we leave the NCM algorithm to
freely choose the variable to be specified, and to set its value, for each flash to be computed.
The graphical abstract (GA) presents, as an illustration of the results that can be obtained, the
computed SLV flash pressure, flash where the vapor phase is incipient, as a function of zCO2 (=
global mole fraction of CO2). T is constant throughout the curve (T = 313.15 K), as it is the
global drug/solvent ratio (= 1.1776 mol acetaminophen/kg ethanol). The computed solid phase
mole fraction decreases as zCO2 decreases, tending to zero as the LSV point is approached. Such
point is a double saturation point where a major L phase is simultaneously at equilibrium with
two incipient phases: a pure-acetaminophen S phase and a V phase. No stability tests were
performed for the computed equilibria. Such tests would identify the valid segments of the
curve shown in the GA. After doing that, the phase diagram would be completed by the addition
of other types of curves, such as the solid-fluid equilibrium curve of incipient S phase. The
NCM was able to properly capture the steep part of the curve in the GA.
Keywords: SLV flash, continuous set, computation, algorithm, numerical continuation method

[1] J.V. Mattos et al. DEQ, UEM, BRASIL.Yet unpublished.

[2] S.B. Rodriguez-Reartes, M. Cismondi, E. Franceschi, M.L. Corazza, J.V. Oliveira, M.S.
Zabaloy. J. Supercrit. Fluids 50 (2009) 193–202.

220
 Upgrading of biolubricants by CO2 technology
Aldana Pizzanoa,b, Sabrina Belén Rodriguez Reartesa,b,c, Pablo Hegela,b *
a
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca, (8000), Argentina
b
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253,
Bahía Blanca, Argentina
c
Department of Chemical Engineering, ETSEQ, Universitat Rovira i Virgili, Avinguda Països
Catalans 26, 43007, Tarragona, Spain

*corresponding author email address: phegel@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Lubricants play a vital role in modern society for the functioning of vehicles and the operation
of industrial units. However, mineral oils have a significant environmental impact, and there is
an increasing use of more eco-friendly options produced from renewable raw materials, which
are less toxic to humans and ecosystems. In this context, the demand for lubricants made from
oils and fats of vegetable and animal origin increases as the trend towards more sustainable
products advances. Biolubricants can be obtained from biodiesel through a reaction with
special tertiary alcohols like trimethylolpropane (TMP). Biolubricants obtained through this
pathway have interesting applications in the steel industry as special hydraulic oils. In the
reaction, fatty acid esters are usually in excess to ensure complete conversion of TMP and the
production of trimethylolpropane trimethyl esters (the desired product). As a result, fatty acid

221
esters remain after the reaction in the final biolubricant product, affecting its viscosity index
and oxidation stability.
In this study, we evaluate the use of liquid and supercritical CO2 for the purification of a
biolubricant obtained through classical esterification of fatty acid methyl esters (FAME) with
TMP at 120 ºC for a 4-hour reaction under vacuum. The product obtained is filtered at 5 ºC to
isolate solids and other undesired products, and the final biolubricant is analyzed to determine
its FAME content. Subsequently, we investigate the purification and upgrading of this
biolubricant using liquid and supercritical CO2 as solvents.
We utilize in this work a high-pressure view cell to examine the phase equilibria of the
biolubricant and CO2, as well as the purification of the biolubricant through a single-stage
contact. We introduce a specific mass of biolubricant (approximately 7 g) and CO2
(approximately 35 g) at room temperature. The temperature of the cell is then set under stirring,
and once the system reaches a stable desired temperature and pressure, we sample the system
to determine the composition of the liquid and solvent phases. Next, we perform a complete
extraction of the heavy liquid phase to assess the characteristics of the final product. The results
demonstrate that both, liquid and supercritical CO2, can be used to extract the remaining FAME
in the biolubricant. Using liquid CO2, up to 3% of FAME can be obtained in the solvent phase,
allowing for the removal of up to 80% of the initial FAME content present in the biolubricant.
Conversely, supercritical CO2 (40 ºC and 150 bar) exhibits a solubility of 8 wt.%, making it
possible to remove 99% of the FAME.

Keywords: Biolubricant, Purification, Supercritical CO2

222
 High-pressure viscosity measurements
of orange oil saturated with CO2
Mariana Fortunatti-Montoyaa,b, Pablo E Hegela,b *, Selva Peredaa,b,c
a
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca, (8000), Argentina
b
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253,
Bahía Blanca, Argentina
c
Thermodynamics Research Unit, School of Engineering, University of KwaZulu Natal, Howard
College Campus, King George V Avenue, Durban 4041, South Africa

*corresponding author email address: phegel@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Thermophysical properties play an important role in the design of packed columns. In
particular, the viscosity of the liquid phase determines the hydraulic behavior of the column
and controls the mass transfer phenomena that govern the separation process. It is well known
that the height of the theoretical stage in the supercritical CO2 fractionation of oily substrates
is highly dependent on the viscosity of the saturated liquid phase. Predicting the viscosity of
liquid mixtures, based on pure compound properties, is a complex challenge. Furthermore,
supercritical extraction processes involve mixing a liquid substrate and supercritical CO2,
which further increases the complexity of accurately modeling this property. Despite this, it is

223
a common practice to assume an ideal solution behavior to estimate the properties of the
raffinate phase in the column, even though it may deviate substantially from the actual mixing
property. Additionally, it is worth noting that viscosity is highly sensitive to the CO2
concentration in the saturated phase.
In this work, we demonstrate the development of a high-pressure viscosimeter for saturated
liquid mixtures under high pressure for the purpose of supercritical process engineering design.
A high-pressure falling weight viscometer is used to measure the viscosity of saturated liquid
mixtures of orange oil + CO2 at different temperatures (293 K to 333 K) and saturated pressures
(20 bar to 95 bar). This technique has been employed by various authors to measure the
viscosity at high-pressure conditions of different oily substrates + CO2 systems. In particular,
we investigate the viscosity behavior of orange oil saturated with CO2 (0 - 62 molar % of CO2
in the liquid phase).
Results show the operating temperature influences the viscosity of both the pure oil and the
CO2-saturated oily substrate. It has been observed that the effect of temperature is more
pronounced in the variability of the viscosity of the pure oil compared to the analysis of the
orange oil + CO2 saturated mixture. Additionally, the viscosity of the liquid mixture decreases
with increasing CO2 concentration at a given operating temperature. This effect is remarkably
noticeable at the lowest temperature studied (293 K). The new viscosity data is properly
correlated using a modified version of the Litovitz equation.

Keywords: Orange oil, Supercritical CO2, Viscosity

224
 Measurement and correlation of PVT data for the
CO2+hydrogen+methanol+water system
Natalia S. Cotabarrena,b, Pablo E Hegela,b *, Francisco A. Sanchez a,b, Selva Peredaa,b,c
a
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca, (8000), Argentina
b
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253,
Bahía Blanca, Argentina
c
Thermodynamics Research Unit, School of Engineering, University of KwaZulu Natal, Howard
College Campus, King George V Avenue, Durban 4041, South Africa

*corresponding author email address: phegel@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
CO2 hydrogenation for the synthesis of methanol (CH3OH) is an interesting route to mitigate
CO2 emissions and promote a sustainable economy considering CO2 capture/utilization and H2
storage. Based on the current industrial methanol production from syngas (H2+CO and minor
quantities of CO2 and CH4), CO2 hydrogenation can be carried out in gas-solid catalytic
reactors at moderated pressures (50 to 100 bar) and temperatures between 490 K and 570 K.

225
However, the current industrial method requires a large recycle flow of syngas due to the
limited conversion achieved in the reactor. An interesting concept is to apply in situ
condensation of methanol or water (subproduct) operating at higher pressures (120 to 500 bar),
which helps to circumvents the costly use of adsorbents or additional coolers to increase the
conversion.
Volumetric properties of reactive systems are needed to elucidate kinetic mechanisms and carry
out a proper design of high-pressure continuous reactors, in particular for reactors with in-situ
condensation of products. To our knowledge, experimental PvT data of the system
(CO2+H2+CH3OH+H2O) have not been reported under the temperature (493.15 K to 563 K)
and pressures (70 to 400 bar) range of interest to carry out the CO2 hydrogenation. Also,
accurately predictive modeling of the volumetric properties and phase behavior of this non-
ideal system, under supercritical conditions, can be complex due to the asymmetric molecular
size and polarity between reactants (CO2/H2) and products (mainly CH3OH and H2O).
In this work, we adapted a high pressure/temperature stainless-steel constant volume cell
(12.76 cc) to experimentally study the pressure-temperature isochoric behavior of quaternary
synthetic mixtures under different stoichiometric molar ratios. The equipment has been
calibrated in the range of operating conditions using pure fluids (methanol, CO2, water). These
measurements were compared to PvT data from the National Institute of Standard and
Technology (NIST). The uncertainty in the density values is about 1.2 % based on calibration
studies.
In this work, we report new experimental PvT data of non-reactive mixtures of
H2+CO2+CH3OH+H2O in the range of temperature and pressure of industrial interest. Mixtures
have been prepared considering initial molar ratios of 1:3 CO2:H2, and different hypothetical
stoichiometric conversions of CO2 (60 % to 94 %) to CH3OH:H2O (1:1). Isochoric studies of
the multicomponent system between 0.13 g/cc and 0.5 g/cc show evidence of the phase
transition, from heterogeneous to homogeneous phase condition at temperatures and pressures
higher than 530 K and 240 bar, respectively. The phase equilibria and PvT data are modeled
using RK-PR, a three-parameter equation of state. We selected the RK-PR because of its
simplicity and proven accuracy to represent volumetric properties of other hydrogenation
reactors.

Keywords: Hydrogenation, Supercritical CO2, Isochores,

226
 Hibiscus sabdariffa L. leaves as an alternative source of bioactive
compounds obtained by supercritical technology
Gabrielli N. Clímacoa,*, Renata Vardanegab, Luiz H. Fasolina
a
University of Campinas, Av. Zeferino Vaz, Campinas-SP, Brazil
b
University of Minho, R. da Universidade, Braga, Portugal
*Corresponding author email address: gabiclimaco7@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
In the last few years, it has been possible to observe a greater demand on the part of consumers,
for healthier foods, rich in bioactive compounds, and that cause minimal impacts on the
environment. Among the plant matrices rich in bioactive compounds is Hibiscus Sabdariffa L.,
known for its calyx and flowers presenting a diversity of compounds beneficial to health.
However, its leaves, in addition to representing the largest part of the plant, are generally
wasted during processing. Therefore, the objective was to extract bioactive compounds from
hibiscus leaves through supercritical fluid extraction (SFE) using water and ethanol as co-
solvents. A complete full-factorial design (2 x 4 x 2) was carried out, varying temperature (40
and 60°C), pressure (100, 175, 250 and 300 bar) and co-solvent proportions (5% and 15%,
w/w). The overall yield (X0), content of bioactive compounds (phenolics, flavanols, tannins)

227
and antioxidant activity were evaluated. The best condition for the extraction of bioactive
compounds was obtained at a temperature of 60 ºC, 100 bar and 15% ethanol, and 40 ºC, 175
bar and 5% water, however, the highest X0 was obtained under maximum conditions, of the 3
factors, for the two co-solvents, being 26.26% and 16.53%, respectively. This shows that
despite the high yield, part of the compounds probably suffered degradation when exposed to
maximum conditions for a long time. For phenolic compounds, values ranging from 6.00 ±
0.01 to 246 ± 3 mg GAE/g of dry extract for extraction with ethanol, and 0.436 ± 0.01 to 3 ±
0.07 mg GAE/g of dry extract for extraction with water were obtained. A similar behavior was
observed for flavanols, with higher values obtained for extraction with ethanol, with a
maximum of 261 ± 3 mg GAE/g of dry extract, and for extraction with water, a maximum of
1.04 ± 0.06 mg GAE/g of dry extract. Regarding tannins, extraction with ethanol was also
superior, with values ranging from 4.80 ± 0.01 to 237 ± 3 mg GAE/g of dry extract, and for
water 0.419 ± 0.00 to 3.010 ± 0.03 mg GAE/g of dry extract. For the antioxidant activity, the
result differed for the two co-solvents used, with a maximum at 40 ºC and 100 bar for ABTS
and 60 ºC and 250 bar for FRAP, both with 5% water, and for ethanol, the best yield was
obtained at 60 ºC and 100 bar, with a proportion of 5% for ABTS and 15% for FRAP. Such
variation between methods can be explained by the fact that some of the compounds present in
the extracts have similar maximum UV wavelengths to the compounds in the reagents.
Comparing the two extractions, it was possible to observe that water allows a superior
extraction for compounds with antioxidant activity, while ethanol performed better for phenolic
compounds, flavonoids and tannins. From these results, it is observed that each solvent has a
unique effect for each compound of interest, and can be influenced both by pressure and by
temperature. In addition, it can be seen that hibiscus leaves are a potential source of bioactive
compounds and that supercritical technology is efficient for obtaining extracts rich in
compounds for the food, pharmaceutical and cosmetic industries.

Keywords: Leaves, hibiscus, supercritical extraction, antioxidant activity,

228
 Solubility of aspirin, ketoprofen and R-(-)-carvone in
supercritical CO2 in binary, ternary and quaternary systems:
Effect of co-solutes

Katia. Dujarrica, Isabela T. Coutinhob, Glaucia T. Mantuanelib, Thierry Tassainga,

Mathilde Champeaub,*
a
Institut des Sciences Moléculaires, UMR 5255 CNRS, University of Bordeaux, 351, Cours de la
Libération, F-33405 Talence Cedex, France
b
Center of Engineering, Modeling and Applied Social Sciences, Federal University of ABC, Santo
André, SP 09210-580, Brazil
*Corresponding authors: mathilde.champeau@ufabc.edu.br

GRAPHICAL ABSTRACT

ABSTRACT

Determining the solubility of compounds in supercritical fluids (SCF) is of primary importance

to understand and control SCF-assisted processes such as extraction, separation or
impregnation. Up to now, most of the studies that have been reported focused on the solubility
of a single compound in SCF (binary systems), and particularly in supercritical carbon dioxide
(scCO2), under different conditions of pressure and temperature. A large set of solubility data
is also available on systems that involve a compound, scCO2 and a co-solvent such as acetone,
methanol, ethanol, with the purpose to enhance compound solubility in the CO2 phase. In
contrast, much less works have been published on the solubility of multicomponent systems
(i.e. mixture of compounds) in scCO2. However, many scCO2-assisted processes involve

229
multicomponent systems instead of a single molecule and would benefit from solubility data
on mixture of compounds.
In this study, to better understand the solubility of multicomponent systems in supercritical
CO2, an in situ IR spectrophotometer coupled to a high-pressure cell was used to measure the
solubility of aspirin, ketoprofen and carvone in binary, ternary, and quaternary systems at 333.2
K and 353.2 K in pressures between 5 and 30 MPa. DSC and ATR-FTIR allowed measuring
the melting temperature of the compounds and optical microscopy was used to observe the
phases. Aspirin formed a eutectic mixture with ketoprofen in the studied proportions, and the
melting point of the mixture is depressed by scCO2. In the case of the ternary mixture with
carvone, the aspirin dissolved so a liquid-vapor equilibrium was established. This resulted in
an enhancement of its solubility in ternary and quaternary systems (graphical abstract), as
shown by the Solubility Enhancement parameter ranging between 30 to 822%. On the contrary,
ketoprofen solubility was higher in binary systems (liquid state), due to a decrease in vapor
pressure when in solution (graphical abstract). Finally, carvone solubility was increased by the
presence of other compounds at 333.2 K and pressures below 9 MPa, whereas at other
conditions, the solubility decreased because of a decrease in vapor pressure.

Keywords: Solubility, thermodynamic measurements, deep eutectic solvent, mixture, enhancer

effect.

230
 Supercritical CO2 extraction of the compounds of the old
cinnamon leaves (Miconia albicans) and antioxidant activity
evaluation
Vicky A. Rambo Ceriolia, Elissandro J. Kleina, Gabrielle C. Peitera, Edson
A. da Silvaa*, Paula A. Palsikowskia, Vladimir Ferreira Cabralb, Lúcio Cardozo
Filhoc
a
Western Paraná State University, Toledo, PR, Brazil
Center for Engineering and Exact Sciences, Western Paraná State University, Toledo, PR,
Brazil.
State University of Maringá – Food Engineering Department, Maringá, PR – Brazil
State University of Maringá – Chemical Engineering Department, Maringá, PR – Brazil
*corresponding author email address: edson.silva2@unioeste.br

GRAPHICAL ABSTRACT

ABSTRACT
Miconia albicans, known as old cinnamon, is a native Brazilian plant used in traditional
medicine as tea. Interest in plants with therapeutic potential has increased, especially in
phenolic compounds, the main group of antioxidant substances in plants. Phenolic compounds
have significant potential for preventing and treating diseases caused by free radicals. Some
studies aimed at describing the biological effects of plant compounds of the genus Miconia and
others aiming to evaluate the total phenolic compounds and antioxidant activity have shown
promising results. There are no reports of studies that evaluated the supercritical extraction
method to obtain compounds of old cinnamon leaves. Thus, the main objective of this study
was to use this alternative technique to evaluate the effects of temperature and pressure on the
yield of oil extracted from supercritical CO2, besides evaluating the content of phenolic

231
compounds and the antioxidant capacity of the extract obtained. This study evaluated the yield
of old cinnamon leaf compounds (Miconia albicans) using supercritical fluid extraction. The
extractions were carried out under the following conditions: temperature of 40, 50, and 60 ºC,
pressure of 140, 170, and 200 bar, and volumetric flow rate of 2.0 cm3.min-1. The extracts were
characterized according to the content of phenolic compounds and antioxidant activity. At
higher temperatures and pressure (200 bar and 60 °C), the highest extraction yield (0.793% by
mass) was obtained, and at lower temperatures and pressure (140 bar and 40 °C), the lowest
extraction yield (0.570% by mass) was obtained. The Sovová model was used to describe the
extraction kinetics. The analysis of the phenolic compounds (Folin-Ciocalteau) resulted in a
value of 29.77 ± 0.25 ìg GAE/g sample. For the study of the antioxidant activity (DPPH), the
value of EC50 was 53.36 ìg.cm-3, the extract being characterized as moderately active. The
results obtained were of great importance since there are no studies in the literature to date of
obtaining compounds of old cinnamon leaves (Miconia albicans) using supercritical fluid
extraction. However, further investment in research and development is still required so that
the extraction conditions are optimized to improve the yield and the profile of compounds
obtained.

Keywords: old cinnamon, supercritical extraction, phenolic compounds, antioxidant activity

232
 A circular economy approach for the processing of fisheries
waste: Pressure-intensified extraction of valuable products from
fish roes
Luis I. Granonea*, Rocío Isla Naveirab,c, Daniela L. Lamasb,c, María A. Fanovichd, María S.
Churioa, Águeda E. Massab,c
a
Departamento de Química y Bioquímica, Facultad de Ciencias Exactas y Naturales, Instituto de
Investigaciones Físicas de Mar del Plata, IFIMAR (CONICET- UNMdP), Deán Funes 3350, 7600
Mar del Plata, Argentina.
b
Instituto Nacional de Investigación y Desarrollo Pesquero, INIDEP, Paseo Victoria Ocampo N° 1,
7600 Mar del Plata, Argentina.
c
Instituto de Investigaciones Marinas y Costeras, IIMyC (CONICET-UNMdP), Rodríguez Peña 4046,
7600 Mar del Plata, Argentina.
d
Instituto de Investigaciones en Ciencia y Tecnología de Materiales, INTEMA (CONICET-UNMdP),
Av. Colón 10850, 7600 Mar del Plata, Argentina.
*corresponding author email address: ligranone@mdp.edu.ar.

GRAPHICAL ABSTRACT

ABSTRACT
The urgent need for environmental protection and human health preservation is forcing
industrial processes to adopt a circular economy scheme. This transformative approach aims to
minimize waste generation, maximize resource efficiency, and foster sustainability. While
advancements in technology have revolutionized many industries, the fishing industry has

233
historically lagged behind in adopting these innovations, particularly in the treatment of by-
products and waste recovery. Currently, fish by-products are utilized in the production of fish
oil, fishmeal, fertilizers, and feed for pets and livestock [1]. These products often have a limited
economic value, which poses challenges in implementing advanced technologies that could
improve their utilization. Fish viscera contain added-value components that are not fully
exploited. In particular, fish roes stand out for their high content of polyunsaturated fatty acids
(PUFAs), especially omega-3 fatty acids, which are highly valuable in the food, cosmetic, and
pharmaceutical industries [2]. Furthermore, the roes contain secondary metabolites that protect
themselves from the substantial exposure to UV rays they suffer in their natural environment.
These remarkable photoprotective attributes are derived mainly from gadusol and gadusol
derivatives, which are highly polar compounds with great potential in the cosmetic industry for
use in sunscreen formulations [2,3]. The conventional methods used to extract these high-
added-value components from fish roes often rely on the use of toxic organic solvents, which
pose risks to both the environment and human health. In addition, these procedures involve
lengthy and energy-intensive separation steps, further adding to their inefficiency. Therefore,
it is crucial to explore efficient and environmentally friendly alternatives. In this context,
extraction and fractionation techniques based on pressure-intensified technologies, particularly
those involving the use of supercritical fluids, emerge as an appealing solution [4].
In this work, omega-3-rich oil was extracted from dried roes of aquacultured Wreckfish
(Polyprion americanus) using supercritical CO2 (scCO2) with and without ethanol as a
cosolvent. At a temperature and pressure of 313 K and 35 MPa, respectively, an extraction
yield of fatty acids of approximately 7.0 % relative to the dry weight of the roes was achieved.
According to the chemical profile, the extracted fatty acids consist of 30.0 wt. % saturated fatty
acids (SFAs), with palmitic acid (C16:0) as the predominant component (61.0 wt. % of total
SFAs), 36.9 wt. % monounsaturated fatty acids (MUFAs), with oleic acid (C18:1n9) as the
main constituent (55.5 wt. % of total MUFAs), and 32.9 wt. % PUFAs, from which omega-3
fatty acids account for 92.1 wt. % and omega-6 fatty acids account for 7.9 wt. %. While DHA
(C22:6n3) and EPA (C20:5n3) were identified as the primary omega-3 fatty acids (85.5 wt. %
and 9.9 wt. % of total omega-3 fatty acids, respectively), linoleic acid (C18:2n6) was
determined to be the predominant omega-6 fatty acid (65.4 wt. % of total omega-6 fatty acids).
Both the yield and chemical profile of the extracted fatty acids remained similar regardless of
the addition of ethanol as a cosolvent. However, when pure scCO2 was used for the extraction,
the highly polar compound gadusol was not extracted, and hence, its concentration in the roes
remained unaltered. In contrast, the addition of ethanol at a cosolvent-to-feed mass ratio of 5
g/g resulted in a 30 wt. % reduction in the gadusol content of the roes after the extraction.
The proposed approaches allow for the efficient extraction of valuable compounds from fish
by-products while minimizing waste generation and environmental impact. Thus, the adoption
of supercritical fluid technologies presents a promising alternative for the fishing industry's
transition toward sustainability.

[1] Cooney, R. et al., J. Clean. Prod. 2023, 392, 136283.

[2] Guedes, M. et al., Separations 2022, 9, 210.
[3] Arbeloa E. M. et al., Food Chem. 2010, 119, 586-591.
[4] Gallego R. et al., Trends Anal. Chem. 2019, 116, 198-213.

Keywords: circular economy, fish roes, scCO2 extraction, omega-3 fatty acids, gadusol.

234
 Use of a pressurized aqueous solution of a natural deep eutectic
solvent to extract bioactive compounds from the Hass avocado
epicarp
Juan F. Grisales-Mejíaa,*, Margarita M. Andrade-Mahechaa, Hugo A. Martínez-Correaa,*, Jose
A. Mendiolac, Alejandro Cifuentesc, Elena Ibañezc
a
Universidad Nacional de Colombia, Carrera 32 # 12 – 00, Palmira, Valle del Cauca, Colombia
c
Institute of Food Science Research - CIAL (CSIC-UAM), Nicolas Cabrera 9, 28049 Madrid, Spain
*corresponding authors email address: jfgrisalesm@unal.edu.co & hmartinezco@unal.edu.co

GRAPHICAL ABSTRACT

ABSTRACT
Extraction processes play an important role in the recovery of bioactive compounds present in
different natural matrices. The above, combined with the need to protect the environment, has
led to the development and implementation of more sustainable processes, with reference to
the Sustainable Development Goals (SDGs), the principles of green chemistry, and the
principles of green engineering. In this context, naturales deep eutectic solvents (NADES),
have become an interesting alternative as they have demonstrated good extraction properties.
Moreover, these novel solvents are synthesized from natural products such as sugars, fatty
acids, quaternary salts, organic acids, polyalcohols, among others, through hydrogen bond
interactions. This is because these solutes can act as both hydrogen bond acceptors (HBA) and
donors (HBD). For this reason, NADES are considered biodegradable, biocompatible, safe,
and cost-effective. However, few studies have combined these solvents with eco-friendly
extraction technologies, such as Pressurized Liquid Extraction (PLE). Therefore, the objective
of this study was to optimize the extraction conditions of the PLE method using NADES based
on choline chloride and fructose (ChCl:Fru) to maximize the presence of phenolic compounds
and the antioxidant capacity of the extracts obtained from Hass avocado peel (Persea
americana Mill.). A response surface design evaluated the effect of water percentages (50%,
70%, and 90%), molar ratios between HBA and HBD (2:1, 1:2, 1:3.5), and extraction
temperature (40 °C, 80 °C, and 120 °C) on total phenol content (TPC), monomeric anthocyanin
content (MAC), and antioxidant capacity against the DPPH radical. The selection of the
NADES was made based on previous results. Their preparation was performed in a water bath
at 80°C with constant agitation until obtaining a transparent and homogeneous solution. The
extraction was carried out at 100 bar and a static time of 20 min. Results showed that the
optimal extraction conditions (desirability level = 0.9183) were 70°C, 60% water, and a 1:1

235
molar ratio. These conditions allowed for obtaining extracts with TPC values of 157.6 mg gallic
acid equivalent/g dry sample, MAC of 7.9 cyanidin 3-glucoside/100g dry sample, and
antioxidant activities of 1378.23 micromol Trolox equivalent/g dry sample. These values were
higher than those estimated for extracts obtained using water at 70°C, 100 bar, and 20 minutes
of extraction. In this latter case, the values were 110.1 mg gallic acid equivalent/g dry sample,
6.3 cyanidin 3-glucoside/100g dry sample, and 1358.3 micromol Trolox equivalent/g dry
sample. These findings evidence the effectiveness and environmentally friendly nature of
employing pressurized aqueous solutions of NADES for extracting bioactive compounds from
Hass avocado epicarp. This opens the possibility for their utilization in developing various
products across pharmaceutical, cosmetic, and food sectors.

Keywords: Green chemistry, neoteric solvents, waste, subcritical water, Persea americana
Mill.

236
 Predictive Correlation for High-Pressure Phase Equilibria of
Nitrogen + n‑Alkane Binary Mixtures with the RKPR EoS
Martina J. Quinzioa,b, Federico E. Benellia,b, Sabrina B. Rodriguez-Reartesc,d,e, Martin
Cismondi Duartea,b,*
a
Universidad Nacional de Córdoba (UNC) - Facultad de Ciencias Exactas, Físicas y Naturales. Av.
Vélez Sarsfield 1611, X5016GCA, Ciudad Universitaria, Córdoba, Argentina.
b
CONICET. Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada
(IPQA), Córdoba, Argentina.
c
Departamento de Ingeniería Química - Universidad Nacional del Sur (UNS), Av. Alem 1253, 8000,
Bahía Blanca, Argentina.
d
Planta Piloto de Ingeniería Química (PLAPIQUI)- Universidad Nacional del Sur (UNS-CONICET),
Camino La Carrindanga Km 7, 8000, Bahía Blanca, Argentina.
e
Department of Chemical Engineering, ETSEQ, Universitat Rovira i Virgili, Av. Països Catalans 19,
43007, Tarragona, Spain.

*corresponding author email address: martin.cismondi@unc.edu.ar

GRAPHICAL ABSTRACT

237
ABSTRACT
Nitrogen (N2) is one of the most common non-hydrocarbon components of natural gas and
oil. It is extensively used in oilfield operations for reservoir pressure maintenance, gas
cycling and gas lift. An increasing number of wells require pressure conservation to maintain
production levels, which can be achieved by injecting natural gas, carbon dioxide or nitrogen.
Nitrogen injection into oil reservoirs is an enhanced oil recovery technique that has several
advantages, as nitrogen is abundant, economically easy to obtain and depending on injection
rate and pressure in the well, the cost of nitrogen may represent up to one quarter to one half
the cost of natural gas.
A thorough understanding of N2 - oil phase behaviour is essential for nitrogen applications in
enhanced oil recovery; for example, the N2 - oil equilibrium phase diagram can be used to
establish miscibility and immiscibility conditions for the mixture.
A correct and consistent description of the phase behaviour for these types of multi-
component systems requires, in the first place, a good representation of the behaviour of the
binary N2 - n-alkane constituent systems. In particular, those systems with a high degree of
asymmetry are more challenging to model.
Predicting the phase behaviour of N2-containing systems with hydrocarbons is a challenging
task because all N2 - n-alkane binary mixtures develop, except with methane, type III phase
diagrams according to the classification scheme of van Konynenburg and Scott. The type III
mixtures present two distinct critical curves, one starting at the critical point of the
component with the highest critical temperature and reaching to infinite pressures, and
another critical curve starting at the critical point of the component with the lowest critical
temperature and meeting a three-phase liquid-liquid-vapour line at a higher temperature final
critical point.
In previous studies of the research group, the superiority of the generalised Redlich-Kwong-
Peng-Robinson equation of state (RKPR EoS) compared to the classical Peng-Robinson (PR
EoS) in predicting the vapour-liquid equilibrium (VLE) of binary n-alkane - n-alkane
systems, and then also for multicomponent mixtures, was demonstrated. The present work
was conceived as a continuation of these previous studies.
With the aim of proposing a model able to correctly and consistently describe the phase and
volume behaviour of N2 - n-alkane binary mixtures, including the most asymmetric ones, we
develop in this work for PR and RKPR EoS new correlations of the attractive parameters kij
with temperature dependence for the homologous series of binary mixtures formed by
nitrogen with normal alkanes. This also implies the parameterisation of pure nitrogen
parameter for RKPR EoS (δ1), whereas for each n-alkane this structural parameter will be
taken from a previous work in which a δ1 correlation for n-alkanes was optimized.
The results show, for both PR and RKPR EoS, a very good predictive power for the phase
behaviour of N2 - n-alkane binary systems. RKPR EoS shows a superior performance
compared to PR EoS which is more noticeable in the case of more asymmetric systems.

Keywords: nitrogen, RK-PR equation of state, n-alkane

238
 Effect of cosolvent (ethanol/water) on the supercritical CO2
extraction of flavonoids from hydroethanolic suspensions of
calafate fruit
Cristian Morenoa, Gonzalo A. Núñeza,*, Laura Cuellarb, José M. del Valleb
a
Department of Chemical and Environmental Engineering, Universidad Técnica Federico Santa
María, Vicuña Mackenna 3939, San Joaquín, Santiago Chile
b
Department of Chemical and Bioprocesses Engineering, Pontificia Universidad Católica de Chile,
Vicuña Mackenna 4860, Macul, Santiago Chile.
*Corresponding author E-mail: gonzalo.nunezm@usm.cl

GRAPHICAL ABSTRACT

ABSTRACT

In recent years, the consumption of berries has gained great importance in promoting population
health due to their high antioxidant content. Berberis microphylla G. Forst (calafate) is a native
plant of the Argentine and Chilean Patagonia, and its fruits are a natural source of flavonoids and
secondary metabolites with antioxidant and anti-inflammatory properties. Supercritical carbon
dioxide (scCO2) extraction is an attractive method for working with plant matrices as it leaves no
solvent residues in the extracts, and it is applicable to solid or liquid substrates. The main limitation
of scCO2 extraction to recover valuable polar compounds from calafate is its low affinity with this
kind of compounds. For this reason, scCO2 can be modified with polar solvents, specifically
mixtures of ethanol and water, to favor the partitioning of the molecules as flavonoids.
The objective of this study was to investigate the effect of the composition and quantity of cosolvent
mixture (ethanol-water) on the recovery of flavonoids from calafate fruits using suspension
extraction with scCO2. To achieve this, an SFE extraction screening unit for processing solid

239
substrates was modified for extracting suspension and it was adapted to allow the addition of
cosolvent under extraction conditions of 50 °C and 30 MPa. Different compositions of ethanol-to-
water will be evaluated to prepare the suspension (69:31, 49:51, 32:68, and 15:85 % w/w) in an
Ultra Turrax equipment, as well as different compositions of the cosolvent (ethanol:water = 86:14,
83:17, 78:22, and 56:44 % w/w). For comparison purposes, milled calafate fruit was extracted with
scCO2 only at the same extraction conditions. Preliminary results show a global extraction yield of
22% and 6% for scCO2 extraction of solid substrate and suspension, respectively. Despite of this,
it is expected that the results will demonstrate higher selectivity towards the target compounds and
improved yields when extracting in suspensions compared to packed bed extraction with scCO2,
due to the cellular fractionation that occurs when mixing the fine substrate in the suspension.
Additionally, it is expected to obtain an extract with high purity surpassing the content of
flavonoids, specifically anthocyanins, in contrast to extracts obtained with conventional methods.

Keywords: Berberis microphylla (Calafate), Flavonoids, hydroethanolic suspension;

supercritical CO2 extraction; ethanol-water cosolvent mixture.

240
 Algorithm for calculating phase diagrams at set initial global
composition involving fluid and solid-solution phases for
multicomponent chemically reactive systems
Matías J. Molinaa,b, Andrés F. Porras-Giraldoa,b, Aldana Pizzanob, S. Belén Rodriguez-
Reartesa,b,c , Marcelo S. Zabaloya,b,*
a
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253, Bahía
Blanca, (8000), Argentina.
b
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS – CONICET), Camino
“La Carrindanga” Km 7, Bahía Blanca, (8000), Argentina.
c
Dpto de Ingeniería Química, ETSEQ, Universitat Rovira i Virgili, Av. Països Catalans 19, (43007),
Tarragona, Spain.
*mzabaloy@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
The goal of this work was to develop an algorithm for computing, over wide ranges of
conditions, phase envelopes and three-phase envelopes for reactive systems including the
possibility of presence, at equilibrium, of solid phases of the solid-solution type. To our
knowledge, algorithms of such sort are not available in the literature. The familiar constant
global composition (z) phase envelope (PE) of a multicomponent non-reactive system shows,
in the pressure (P) versus temperature (T) plane, a number of curves, such as bubble point, dew
point, liquid-liquid and/or solid (S) - fluid (F) (S+F) curves (z is a vector of global component
mole fractions). In a given point of the PE (which often includes critical points), a phase of
finite size (major phase) of composition z is at equilibrium with a phase of differential size
(incipient phase) of composition generally different from z. The PE is the boundary between
the homogeneous region and the two-phase (heterogeneous) region. The information on the
phase behavior of the multicomponent system of interest becomes more complete if some

241
additional types of lines are plotted within the heterogeneous region, e.g., the three-phase
envelopes (3PEs) or three-phase lines (3PLs, for the case of binary systems). A 3PE is the
boundary between a two-phase region and a three-phase region. In a point of a 3PE, two phases
of finite size are at equilibrium with an incipient phase (still being the global composition equal
to z). The set of all PE segments plus all the auxiliary lines (e.g., 3PEs or 3PLs) constitute a
quite complete constant z diagram which is named “isopleth” (IP). For reactive systems, the
global composition z changes along the reactive PE (R-PE) and also along the reactive 3PE (R-
3PE) or reactive 3PL (R-3PL) (depending on the degrees of freedom (DsOF) of the reactive
three-phase system). Each point of any of these curves corresponds to the simultaneous phase
and chemical equilibria. Both, in IPs and in reactive IPs (R-IPs) the global mole fractions of
the atoms indeed remain constant, while the global mole fractions of the components are
constant only for the non-reactive IPs. In a previous work [1], calculation algorithms were
developed for all types of lines present in R-IPs involving only fluid phases. In the present
work, the possibility of precipitation of solid solutions has been incorporated to the computed
R-IPs. The modelling approach proposed by Porras et al. [2] was used to represent the
thermodynamic properties of multi-component solid phases (Solid Solution Approach (SSA)).
As case study, we have chosen the carbon dioxide (CO2) + 1,2-propylene oxide (PO) +
propylene carbonate (PC) system, in which chemical bonds are broken or formed as prescribed
by the chemical reaction CO2 + 1,2-propylene oxide ⟷ propylene carbonate. The fluid-state
volumetric and phase behavior of this system was modelled through the well-known SRK-EoS
coupled to quadratic mixing rules (QMRs). The EoS pure component parameter values, and
the values for the binary interaction parameters (considered equal for both the fluid and solid
phases), used in the calculations, are given in ref [3], together with the required “standard
state”-related parameters. Other pure component parameters appearing in the solid-solution
global fugacity expression were obtained, either from databases, or from reproducing the
experimental pure component solid-liquid equilibrium curves. For the chosen reactive system,
complete reactive three-phase line segments and reactive phase envelope segments, including
the possibility of presence of solid phases that are solid solutions, were computed over wide
ranges of conditions using numerical continuation methods. The results are illustrated through
the phase diagram, computed for the initial global composition z0CO2 =0.75, z0PO =0.25 and
z0PC =0.0, which is shown in the Graphical Abstract (GA). The obtained results imply the
existence of complex patterns of behavior for the computed R-IPs (see insert in GA). The
circular marker in the GA is a reactive critical point. Furthermore, a predicted homogeneous
solid-solution region is shown in the pressure-temperature plane in the GA. The presence of
homogeneous-solid regions in mixture isopleths is of impossible prediction by models that
allow precipitation only of solid phases made of pure components (i.e., models that exclude
the possibility of solid solutions). The proposed algorithms have been found to be robust and
this is ascribed to the applied numerical continuation method.
Keywords: Reactive Systems, Chemical and Phase Equilibrium, Algorithm, Solid Solution.
[1] M.J. Molina, S.B. Rodríguez-Reartes, M.S. Zabaloy. Computation of Complete Fluid State
Phase Diagrams at Set Initial Global Composition for Multicomponent Chemically Reactive
Systems. Conference Abstracts Book, p. 136. PetroPhase2021, June 7-9, 2021 (Virtual). [2]
A.F. Porras Giraldo, S.B. Rodriguez-Reartes, M.S. Zabaloy, Fluid Phase Equilib. 547 (2021)
113174. [3] M.J. Molina, S.B. Rodriguez-Reartes, M.S. Zabaloy, Computation and Analysis
of Reactive Isopleths Involving Fluid Phases. Submitted to Fluid Phase Equilib., march 16th,
2023.

242
 Determination of process conditions to obtain Macaúba
(Acrocomia aculeata) oil using Supercritical CO2 and its potential
for application in new products.
Giselle C. M. Airesa, Kelly R. P. Pantojaa, Jean M. L. Pinheiro, Verônica M. S. Bezerraa,
Maria E. F. de Carvalhob, Eduardo G. O. Menezesc, Raul N. de Carvalho Juniora*
a
Federal University of Pará, Rua augusto Corrêa w/n, 66075-900. Belém-Pará. Brazil
b
Federal Rural University of The Amazon, Rua augusto Corrêa Belém-Pará. Brazil
c
Federal Institute of Education, Science and Technology of Rondônia, Rondônia-Pará. Brazil
*corresponding author email address: gisellecsmelo@yahoo.com.br

GRAPHICAL ABSTRACT

ABSTRACT
Acrocomia aculeata is a palm tree native to the Amazon, found throughout Brazil. Its fruit is a
rich source of nutrients, including carbohydrates, fiber, protein, vitamins, and minerals. In
addition, the pulp of the macaúba fruit contains bioactive compounds, such as unsaturated fatty
acids, tocotrienols, carotenoids, phenolics and flavonoids, which give the oil antioxidant, anti-
inflammatory, antidiabetic, antimicrobial properties, as well as photoprotective effect. These
properties present in the pulp of macaúba have been the subject of several studies that show
the potential of this raw material for use in the food, pharmaceutical, cosmetic and biofuels
industries. In this context, the present study aimed to determine the process conditions for
obtaining oil from macaúba pulp through the use by supercritical CO2 as a solvent, exploring
the funtional potential of this extract. Thus, the oil was obtained from the pulp of freeze-dried
macaúba (Acrocomia aculeata). The conditions adopted for extraction were, temperatures of
40 and 50ºC and pressures of 200, 300 and 400 bar, and presenting solvent densities ranging
between 784.29 and 956.07kg/m³, for 90 minutes. The fatty acid profiles of the CO2-SFE
extracts were determined on gas chromatograph (model GC-2010, Shimadzu, Japan) equipped

243
with a flame ionization detector, and a capillary column TG-WAX MS A (30 m·3.20×10−4
m·2.50×10−7 m). The β-carotene contents were quantified in petroleum ether using a
spectrophotometer (model Evolution 60, Thermo Scientific, USA). The extracts of macaúba
obtained presented, on average, 64.12% of oleic acid (n9) and β-carotene with contents ranging
between 12.78 and 84.39μg/g in their composition. The maximum oil yield was obtained at
40ºC and 400bar (21.93 ± 0.13%) in db, generating an extract with a higher concentration of
β-carotene (84.39 ± 0.023 μg/g). The results suggest that the oil of the macaúba pulp presents
high nutritional potential.

Keywords: Macaúba, functional foods, supercritical CO2, bioactive compounds.

244
Subcritical fluid extraction of phenolic compounds: an alternative
to add value to pecan nutshells
Andrea C. Rodríguez-Ruiza*, Miguel E. Cardona Jimenezb, Alexis R. Veleza, Lina M.
Agudelo-Laverdec, Patricio R. Santagapitab, Romina M. Bodoirad
a
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA) –
CONICET – UNC, Av. Vélez Sarsfield 1611, Córdoba, Argentina.
b
Centro de Investigación en Hidratos de Carbono (CIHIDECAR) - CONICET – UBA, Intendente
Güiraldes 2160, Buenos Aires, Argentina
c
Universidad del Quindío. Facultad de Ciencias Agroindustriales. Carrera 15 calle 12 Norte, Armenia,
Colombia.
d
Instituto de Ciencia y tecnología de los Alimentos Córdoba (ICYTAC) – CONICET – UNC, Av.
Juan Filloy S/N, Córdoba, Argentina
*acrodruiz@mi.unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Changes in food trends such as the enhancement of consumer’s awareness regarding the
nutritional properties of food products, have caused an increase in the consumption of nuts,
particularly pecan nuts. According to data from the Pecan Cluster in Argentina, pecan nut
production [Carya illinoinensis (Wangenh) K.Koch], is estimated to be over 2000 tons per
year. The shell, which is produced during walnuts shelling, is equivalent to 40-50% of the
waste generated by this productive system, and it is only used for energy generation.
Many authors have reported the presence of bioactive components in pecan nutshells, including
phenolic compounds with antioxidant, antiradical, or chelating capacity. This has generated a
growing interest in studying this by-product and determining the best extraction method for
recovering these compounds. Among the green techniques, subcritical fluid extraction stands

245
out utilizing water or ethanol + water mixtures at temperatures between 100 and 374 °C, and
pressures above atmospheric to maintain the solvent in a liquid state. It is a safe and fast
technique that achieves high extraction yields of medium and high polarity compounds from a
solid matrix.
Considering these aspects, the use of water + ethanol mixtures at subcritical conditions for the
extraction of phenolic compounds from pecan nutshells was evaluated in the present work. The
pecan nutshell obtained as result of the peeling process was donated by Producor S.A (San
Jerónimo, Santa Fe, Argentina). They were washed, disinfected, dried, milled and sifted before
extraction. The pecan nutshell powder (PNSP) was subjected to different extraction conditions.
The studied variables were temperature (T: 100, 140, 180, and 220°C), ethanol composition in
the solvent mixture (S: 0, 25, 50, 75, and 100 %w/w), and solvent mass flow rate (F: 2.5, 5.0,
and 7.5 g/min). The total phenolic content (TPC) expressed as mg gallic acid equivalents
(GAE)/ g dry PNSP, and antioxidant activity by FRAP and ABTS methods expressed as mmol
Trolox equivalents (Tx eq)/ g dry PNSP of each extract were measured.
The TPC of the extracts varied between 165.93 and 454.61 mg GAE/g dry PNSP, and the
antioxidant activity with a range between 2.41 to 7.63 mmol Tx eq/g dry PNSP, and 1.92 to
5.50 mmol Tx eq/g dry PNSP for FRAP an ABTS, respectively. It was observed that increasing
the extraction temperature enhances the recovery of phenolic compounds, while the opposite
effect is observed with an increase in the ethanol content in the solvent mixture, which leads to
a reduction in TPC values and antioxidant activities. Thus, regarding the bioactive content, the
best extraction conditions were T 200°C, F 2.5 g/min, and S 50% w/w of ethanol.
The results showed that the selected variables influenced the extraction of total phenolic
compounds. The ethanol + water subcritical extraction is an alternative for recovering phenolic
compounds and adding value to pecan nutshells.

Keywords: Subcritical fluid extraction, bioactive compounds, pecan nutshells, antioxidant

activity.

246
 Supercritical fluid extraction of antioxidants from
“Lorena” avocado seeds
Juan Camilo de la Cruz, Luisa Salazar, Isabel Mejía, Gustavo Bolaños*

Laboratory of Applied Thermodynamics and Supercritical Fluids

School of Chemical Engineering, Universidad del Valle
CL 13 100-00 Bldg E30, Cali, Colombia

*corresponding author: gustavo.bolanos@correounivalle.edu.co

GRAPHICAL ABSTRACT

Cryomilling SCFE
50 to 70 ºC Antioxidant test
Freeze drying (DPPH)
12 to 34 MPa

ABSTRACT
From the nine avocado varieties that are produced in Colombia (Lorena, Trinidad, Booth-8,
Fuerte, Hass, Trapo, Santana, Colinred and Ettinger), the “Lorena” variety is one of the most
important for internal consumption. The seeds of this variety are known to contain larger
amounts of phenolic compounds (up to 36% more) than other varieties, in particular the Hass
variety, which is the one that has received a great deal of attention because of its international
demand. The phenolic compounds are in turn known for their biological activity as
antioxidants, and have many potential applications in the food, cosmetics, and nutraceutical
industries.
In this work, antioxidant extracts were obtained from “Lorena” avocado seeds by supercritical
fluid extraction. First, a large batch of fresh seeds was obtained from a local supplier and was
subjected to cryogenic milling up to an average particle diameter of 1.50 ± 0.41 mm. Then, the
pulverized seeds were freeze dried and stored at 4 ºC in a dark and oxygen-free environment
before extraction with supercritical carbon dioxide. Careful manipulation of the avocado seeds

247
was given during the entire processing to avoid the deleterious effect that the polyphenol-
oxidase enzyme has on the phenolic compounds. In fact, this enzyme is present in the seeds
and is known to be responsible for the browning that develops once the seed is mechanically
hurt in the presence of water and oxygen. Such color change is due to oxidation of phenolic
compounds. Thus, maintaining the material in an oxygen-free ambient before freeze drying is
essential for a successful extraction of phenolics.
The milled and dried seeds were subjected to supercritical extraction with CO2 according to a
factorial experiment in which the effect of temperature (from 40 to 50 ºC) and CO2 density
(from 0.7 to 0.9 g/mL) on the yield and antioxidant capacity of the extracts was determined.
Extraction yields were gravimetrically measured, and the antioxidant capacity was measured
according to the well-established 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical
methodology.
Results show that extraction yields up to 5.9% mass of extract/mass of dryed seed are obtained
and the extracts exhibit a significant antioxidant activity, with a concentration that halves the
DPPH free radical (EC50) as low as 4.5 g extract/g DPPH, which compares well with the value
for butylhydroxytoluene (0.13 g BHT/g DPPH), a well-known synthetic antioxidant, given the
fact that no further purification of the avocado seed extracts has been performed. These results
indicate that supercritical fluid extraction of antioxidants from avocado seeds might be a way
to add significant value to a byproduct that today is not used, by producing important
compounds for the food, cosmetics, and nutraceutical industries.

Keywords: Avocado, supercritical extraction, antioxidant

248
 Supercritical fluid extraction from bark of mururé (Brosimum
acutifolium): Sequential extraction study, Chemical composition,
total phenolic compounds, and antioxidant activity
Kelly Roberta P. Pantojaa*, Fernando de Freitas M. de Azevedoa, Eduardo
Gama O. Menezesb, Heloisa L. de Araujoa, Wandson Braamcamp de S.
Pinheiroa, Raul N. de Carvalho Júniora

a
Federal University of Pará, Augusto Corrêa 01, Belém, Brasil
b
Federal Institute of Education, Science and Technology of Rondônia, Governor Jorge Teixeira 3146
Sector - Industrial, Porto Velho, Brasil
*Corresponding author email:kellyramalhosousa@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
The mururé (Brosimum acutifolium) is a tree native to Brazil. Its bark is popularly used in the
Amazon region to treat inflammation and rheumatism, being used in the form of teas and
alcoholic macerations. This work studies the influence of the operating conditions of
temperature (40ºC-60ºC), pressure (250, 300, 350 bar) and addition of cosolvents, in the
extracts obtained by sequential extraction by supercritical fluid using the systems: CO 2, CO2 +
Ethyl Acetate and CO2 + Ethanol in relation to the mass yield, chemical composition, content
of phenolic compounds and antioxidant activity. The qualitative phytochemical analyses were

249
performed by High Performance Thin Layer Chromatography (HPTLC), for the content of
phenolic compounds the Folin-Ciocalteu method was used, and the antioxidant activity was
determined by the DPPH method. For all systems, 10 g of raw material was used. The isotherms
of the system that uses only supercritical CO2 were not possible to be obtained because they
presented non-quantifiable amounts of lipids, a fact that had already been observed during the
centesimal characterization stage of the raw material. In the CO2 + Ethyl Acetate system, for
both isotherms at 40ºC and 60ºC, the pressure increase favored the process yield. Isobarically,
it was possible to notice that the temperature increases also influenced the observed yield, the
highest yield in this system for each isotherm was at 40°C/350 bar (0.46% ± 0.03) and at
60ºC/350 bar (0.80% ± 2.64). For the CO2+ Ethanol system, in both isotherms at 40ºC and
60ºC, the increase in pressure caused a reduction in the mass yield of the extraction. When
evaluating the isotherms isobarically, at 250 bar the increase in temperature causes a reduction
in the extraction yield, however at 300 bar there is an inflection point of the isotherms, from
this pressure the highest yields are obtained at 60ºC, probably by the dominant effect of the
vapor pressure of the solute, the highest yields obtained were at 40°C/250 bar (0.31%±0.03)
and at 60°C/250 bar (0.23%±0.03). The phytochemical screening revealed the presence of
phenolic compounds in the extracts obtained with the use of Ethyl Acetate and Ethanol, being
the solvent Ethyl Acetate more selective for coumarin extraction in the conditions of 40ºC and
60ºC both at 350 bar. The presence of fatty acids in the systems with ethyl acetate and ethanol
was observed, which must have occurred due to the constant flow of CO 2 throughout the
extraction process. The analysis of the content of phenolic compounds and antioxidant activity
was performed in the extracts with higher yields. In this sense, in the CO2 + Ethyl Acetate
system, the extracts were evaluated under the conditions of 40ºC/350 bar and 60º/350 bar,
which presented phenolic compounds content equal to 22.48±0.28 mg GAE/g and 19.43±0.57
mg GAE/g, respectively, evidencing that the temperature causes reduction of these metabolites.
The antioxidant activity expressed values equal to 34.57±0.59 μmol Trolox eq./g (40ºC/350
bar) and 75.67±1.22 μmol Trolox eq./g (60º/350 bar), in which case it was possible to observe
the strong influence of the temperature increase on the antioxidant potential of these extracts.
For the CO2+Ethanol system, under the conditions of 40ºC/250 bar and 60ºC/250 bar, the
contents of phenolic compounds were equal to 171.9±2.48 mg GAE/g and 77.68±1.00 mg
GAE/g, respectively. In this system, the increase in temperature caused a reduction of
approximately 45 % in the phenolic content of the extracts. The evaluation of the antioxidant
activity showed values equal to 59.35±2.93 μmol Trolox eq./g (40ºC/250 bar) and 34.57±2.35
μmol Trolox eq./g (60ºC/250 bar), indicating that the increase in temperature negatively
influenced the antioxidant potential of the extracts studied. In this research, it is observed that
the relationship between the content of phenolic compounds and the antioxidant activity in the
evaluated conditions is not very well established and may be influenced by factors such as the
synergy between the substances, phenolic composition, and the method of evaluation of
antioxidant activity adopted. Thus, the study conducted with mururé demonstrated that the
technology with supercritical carbon dioxide was efficient, considering that the yields obtained
in this work exceeded those obtained by other authors (0.1 to 0.8%) who used from 1.5 kg to
2.5 kg of raw material in maceration extractions. In addition, it was possible to obtain plant
extracts with different chemical profiles, evidencing the selectivity of the studied process.

Keywords: Sequential extraction, mururé, chemical composition.

250
INFLUENCE OF BED FILLING ON THE KINETICS OF THE
EXTRACTION OF PHENOLIC COMPOUNDS FROM
ARAÇAUNA (Psidium myrtoides) USING PRESSURIZED
LIQUIDS
Yaneth M. Monroya, Sara Fragaa, Patricia Berillib, Mario R. Marósquita Jr.b,
Marili Villa Nova Rodriguesc, Daniel Gonçalvesd, Klicia A. Sampaioa
a
Department of Food Engineering, University of Campinas - UNICAMP, 13083-862 Campinas, SP,
Brazil
b
Department of Food Science and Nutrition, University of Campinas - UNICAMP, 13083-862
Campinas, SP, Brazil
c
Chemical, Biological and Agricultural Pluridisciplinary Research Center (CPQBA), University of
Campinas - UNICAMP, 13083-970 Campinas, SP, Brazil
d
Laboratory of Food Technology, State University of Northen Rio de Janeiro – UENF, 28013-602
Campos dos Goytacazes, RJ, Brazil
*corresponding author email address: klicias@unicamp.br yanethmchac@gmail.com
mmarosti@unicamp.br

GRAPHICAL ABSTRACT

251
ABSTRACT

Filling the extraction cell with raw material using pressurized liquids is important for achieving
a high yield of phenolic compounds from araçauna (Psidium myrtoides). The extraction cell,
also known as an extraction vessel or column, is a vessel designed to hold the raw material and
allow for efficient contact between the liquid and the material. It typically features an inlet for
the pressurized liquid and an outlet for the extracted solution. The raw material may be in the
form of solid particles, granules, or a packed bed of material. A systematic study is needed to
select suitable criteria for filling the extraction cell to obtain extraction curves with a high yield.
In the present work, an extraction cell with a volume of 47 cm3 was used, In this cell, different
filling positions of the raw material (RM) were used with an average particle diameter of 0.372
± 0.01 mm and a real density of 1.52 ± 0.01 g/cm3, and glass spheres (GB) with a diameter of
4.5 ± 0,5 mm were also used. The amounts of RM and GB for filling were ~7g and ~47g
respectively in the extraction cell, the following positions were used: (I) at the beginning with
RM and completed with GB, (II) in the middle with RM, it was placed before and then GB,
(III) at the end, first the GB were placed, completing with RM in the extraction cell, and (IV)
a mixture of the RM and GB. In all forms of extraction, 200 bar and 323 K were used at a flow
rate of 1.5 mL/min of 70% ethanol absolute at pH = 2 as solvent. The analysis of the results
showed that when we filled in at the end of the extraction cell, we opted for higher extraction
yields with 43.7 % and more concentrated extracts of total monomeric anthocyanins with 4.55
± 0.124mg/g dry extract. Thus demonstrating that when employing filling techniques in the
extraction cell, knowing the nature of the raw material and the use of pressurized liquids help
to maximize the efficiency of the extraction, promoting uniform contact and optimizing the
separation of the desired components of the raw material.

Keywords: Brazil's Atlantic Forest, Araçauna, Pressurized liquid, Extraction yield.

252
 Development of Deep Eutectic Solvents based on amino acids for
CO2 absorption
Diego Miranda de S. C. a,*, Gabriel S. Santos a, Reinaldo C. Bazito a

Fundamental Chemistry Department, Institue of Chemistry, University of São Pauloa

Av. Prof. Lineu Prestes,748, São Paulo, Brazil
*diego.miranda.costa@usp.br

GRAPHICAL ABSTRACT

ABSTRACT
Global climate change due to greenhouse gas emissions has been intensifying in recent decades
and, as a consequence, the focus of much research is aimed at reducing or capturing CO2
emissions. An interesting way to alleviate this problem is to invest in the reuse or capture of
CO2 using economically viable processes, so the interest in studies of new technologies for
capturing this gas is growing. Among the available technologies, capture by
sorption/desorption, either physical (physisorption) or chemical (chemisorption), has been
used. Chemisorption with organic compounds containing amine groups is the most used today
but its use leads to a large consumption of energy in the desorption process and the use of
volatile amines that make the processes more expensive and environmentally unsound.
Studies with IL (ionic liquids) have brought good results as an alternative to this problem,
because they can present good solubility of CO2, are less volatile and more stable in a wide
range of temperatures and pressures, can also be developed with different combinations of
cations and anions for specific applications. One of the main challenges to make them viable
is to develop materials with lower viscosity and that maintain their sorption capability even in

253
media with a higher amount of water. Another class of materials that has brought a lot of
interest is the Deep Eutectic Solvents (DES), a mixture of a hydrogen bond donor (HBD)
compound with a hydrogen bond acceptor (HBA) compound, resulting in a material that melts
bellow room temperature and, depending on compounds used, has the capability of CO2
sorption. They have several advantages over Ils, such as not demanding elaborated synthetic
and purification steps, lower toxicity, better biodegradability and lower heat of regeneration
(desorption). DES are characterized by not necessarily presenting ionic groups like ILs.
In order to obtain new materials for carbon dioxide capture and better understand their
properties, new DES were developed using greener compounds and with less environmental
impact, using amino acids and polyols as HBD, in conjugation with choline chloride as HBA.
The polyols developed in this work are hyperbranched polyol-polesthers, In order to obtain
DES with a higher CO2 sorption capability, we also synthesized hyperbranched polyesters or
oligomers modified with amines and amino acids such as arginine and proline in their terminal
groups. These were used as HDBs in the obtention of new DES.
All the compounds synthesized were characterized by the usual techniques (H1 and C13 NMR,
MALDI-TOF). The physicochemical properties of the DES obtained were characterized by
techniques such as DSC, NMR and Karl Fischer. We also studied its interaction with CO2
under low and high CO2 pressure conditions, as well as its phase behavior under supercritical
conditions. The results obtained demonstrate that the DES studied show good CO2 sorption
capabilities.

Keywords: CO2, DES (Deep eutectic solvents), Amino Acids

Acknowledgements :
We gratefully acknowledge the support of the RCGI – Research Centre for Greenhouse Gas
Innovation, hosted by the University of São Paulo (USP) and sponsored by FAPESP – São
Paulo Research Foundation (2020/15230-5; 2014/50279-4) and Shell Brasil, and the strategic
importance of the support given by ANP (Brazil’s National Oil, Natural Gas and Biofuels
Agency) through the R&DI levy regulation.

254
 COMPOSITIONS OF FATTY ACIDS AND TOCOPHEROLS
FROM MACAUBA PULP (Acrocomia aculeata) EXTRACTED
WITH SUPERCRITICAL FLUID
Yaneth M. Monroya, Gabriel S. L. Pereiraa, Carlos A. Colombob, Klicia A.
Sampaioa
a
Department of Food Engineering, University of Campinas - UNICAMP, 13083-862 Campinas, SP,
Brazil
b
Center for Genetic Resources, Agronomic Institute of Campinas - IAC, 13020-902 Campinas SP,
Brazil
*corresponding author email address: klicias@unicamp.br, yanethmchac@gmail.com,

GRAPHICAL ABSTRACT

ABSTRACT

The Acrocomia genus, commonly referred to as macaw palm or macaúba, is found in tropical
regions of the Americas, specifically in savanna areas and open forests. Their distribution
extends from Central to southern South America. These palm trees play a significant role in
ecosystems and local economies, and they possess substantial potential for biofuel production
and vegetable oil extraction. The oil can be extracted using different technologies at high or
low pressure, however extraction using supercritical CO2 allows obtaining clean extracts with
high extraction yields and bioactive compounds. In this work, Macauba pulp oil was extracted

255
using supercritical CO2 (scCO2) as the solvent in a single step under conditions of 323K and
300 bar with an average solvent flow rate of 1.5 L/min (2.475 g/min). The scCO2 flowed
through a fixed bed containing approximately 7.0 g of sample for an average total time of
approximately 1 hour and 30 minutes. The fatty acid profile was determined by gas
chromatography, for the identification of the peaks, the retention times of the fatty acid methyl
ester standards were compared, The separation of tocopherols and tocotrienols was
performedby liquid chromatography using methanol: water: ammoniumhydroxide
(99:1:0.1v/v/v) as mobile phase A and pure isopropanol as mobile phase B. Getting high
extraction yields were with 41.56%, the fatty acid composition of the oils obtained from the
pulp of A. aculeata obtained showed oleic acid (56.06%) was the major constituent. CO2 was
efficient in removing tocopherols from macaw palm pulp, among which α-tocotrienol (55.66
µg/g) and β/γ-tocotrienol (63,63 µg/g) were the major constituent.

Keywords: Macaw, Vegetable oil, Fatty acid, Tocopherol, Tocotrienol.

256
 Supercritical technology applied to obtain tucumã oil
(Astrocaryum vulgare): Kinetic parameters and chemical
composition
Rayelle M. Moraisa*, Eduardo G. O. Menezesb, Nayara J. N. da Silvac , Maria C. R. Ferreira
c
, Kelly R. P. de Sousac, Fernando de F. M. de Azevedoc , Carolina B. G. Telesd , Raul N. de
Carvalho Júnior a, c*
a
Graduate Program in Chemical Engineering, PPGEQ/ITEC/UFPA, Federal University of
Pará, Rua Augusto Corrêa S/N, Guamá, 66075-900 Belém, Pará, Brazil.
b
Federal Institute of Education, Science and Technology of Rondônia, IFRO, Porto Velho,
Rondônia, Brazil.
c
Laboratory of Extraction (LABEX), Federal University of Pará, Rua Augusto Corrêa S/N,
Guamá, 66075-900 Belém, Pará, Brazil.
d
Oswaldo Cruz Fundation of Rondônia, FIOCRUZ-RO, Porto Velho, Rondônia, Brazil.
*corresponding author email address: rayellemorais@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
The technology of extraction with supercritical carbon dioxide is highly efficient for obtaining
vegetable oils, although there are still few studies on its application for obtaining oil from
tucumã-do-Pará (Astrocaryum vulgare). The diversity of bioactive substances in tucumã oil
confers quality for this oil to be inserted as an input in the production of products with high
added value of interest to the industry that aim at the formulation of new functional products.
Therefore, the objective of this work was to evaluate the sequential extraction of tucumã-do-
Pará oil via supercritical carbon dioxide, followed by obtaining an ethanolic extract. The
extractions were carried out under conditions of 40°C and 400 bar, and the ethanolic extracts
obtained from concentrations of 5 and 15%. Thermodynamic solubility and kinetic parameters
were estimated using cubic equations of state and mass transfer fundamentals, respectively.

257
The results obtained showed a yield of 38.9% for the oil concentrated in carotenoids
(618.01±4.17µg/g oil). For ethanolic extracts, the highest yield was found at 15% (5.81%) with
antioxidant activity of 315.72±31.09 g extract/g DPPH. The solubility was estimated for two
systems, considering a multicomponent system consisting of fatty acids, the solubility value is
equal to 0.0105 kg oil/kg CO2 and considering a multicomponent system consisting of major
triglycerides, the solubility value is equal to 0.0197 kg oil/kg CO2. Sovová's model presented
a good fit to evaluate the extraction kinetic parameters (RSS<0.01). In a comparative analysis,
it was observed that the addition of ethanol from 5% to 15% caused an increase in the external
mass transfer coefficient (kf) from 4.84 x10-4 to 6.57 x 10-4, confirming the increase in the
power of solubilization. Therefore, the supercritical technology proved to be efficient to obtain
tucumã-do-Pará oil concentrated in carotenoids and the use of ethanol as a co-solvent allowed
obtaining an extract with high antioxidant capacity.
Keywords: Extraction, Supercritical Fluid, Tucumã-do-Pará Oil.

258
 Physical-chemical studies of araçauna flour (Psidium myrtoides)
and extraction yield obtained by sequential extraction in fixed bed
with SFE and PLE technology
Sara Fragaa, Yaneth M. Monroya, Patrícia Berillib, Mário R. Maróstica Jr.b,
Daniel Gonçalvesc, Klicia Araujo Sampaioa
a
Department of Food Engineering, University of Campinas - UNICAMP, 13083-862 Campinas, SP,
Brazil
b
Department of Food Science and Nutrition, University of Campinas - UNICAMP, 13083-862
Campinas, SP, Brazil
c
Laboratory of Food Technology, State University of Northen Rio de Janeiro – UENF, 28013-602
Campos dos Goytacazes, RJ, Brazil
*Corresponding author email address: klicias@unicamp.br, sarafraga11@gmail.com,
yanethmchac@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT
Native fruit species from the Atlantic Forest have a known potential for their application as
food. Some native fruits are commercially consumed on a large scale and have an important
nutritional composition, such as cashew (Anacardium occidentale), pineapple (Ananas
comosus), passion fruit (Passiflora spp.), and jabuticaba (Plinia cauliflora). However, there
are several other fruits that are only known in restricted geographic areas and still being little

259
studied and explored, as the most of red fruits found in the Atlantic Forest. This is the case of
araçauna or purple-araçá (Psidium myrtoides), a fruit with purple skin, dark pink pulp, sweet,
and rich in anthocyanins and other phenolic compounds. This study aimed to evaluate the
lyophilized araçauna pulp flour in terms of its physical-chemical composition and to obtain
extracts from sequential extractions in a fixed bed at 50 °C and 300 Bar, using as extractor
fluids the supercritical CO2 (ScCO2) and pressurized liquids (PLE) ethanol and water, in a
sequence. Fruits with skins and without seeds were lyophilized, crushed, standardized,
submitted to physical-chemical evaluations and to sequential extraction kinetic assays. The
material’s physical properties were: 1.52 g/cm³ real density, 0.39 g/cm³ apparent density, 0.74
porosity, and 177.34 µm mean particle diameter. The chemical evaluation showed 5.78% total
volatile fraction, 4.82%, moisture, 13.9% soluble solids, 3.88% ashes, 1.60% lipids, 4.11%
proteins, and pH 2.1. Results revealed that the extraction kinetics had an optimal time of 270
min, an average accumulated yield of 37.5%, distributed in 0.35% for ScCO2, 19.2% for
ethanol, and 17.9% for water fractions. The lyophilized araçauna flour has potential of
application in nutraceutical foods and the extracts exhibited good recovery yields, being the
ethanolic step best way to extract compounds with an incidence of color as flavonoids.

Keywords: Brazil's Atlantic Forest, purple-araçá, proteins, Extraction yield.

260
Determination of process parameters and antioxidant potential of
Calycolpus goetheanus (O. Berg) essential oil obtained by
supercritical CO2 extraction
Rafael V. Motaa, Nayara J. N. da Silvaa, Eduardo G. O. Menezesb, Maria Eduarda F. de
Carvalhoc, Jean M. L. Pinheiroa, Eloisa H. de A. Andraded, Raul N. de Carvalho Juniora,*
a
Program of Post-graduation in Food Science and Technology, PPGCTA, Federal University of Pará,
Belém, Brazil
b
Federal Institute of Education, Science and Technology of Rondonia, IFRO, Porto Velho, Rondonia,
Brazil
c
Socio Environmental and Water Resources Institute, ISARAH, Federal Rural University of the
Amazon, Belém, Pará, Brazil
d
Adolpho Ducke Laboratory, Botany Coordination, Museu Paraense Emílio Goeldi, Belém, Pará,
Brazil
*corresponding author email address: raulncj@ufpa.br.

GRAPHICAL ABSTRACT

ABSTRACT
The Amazon region stands out for harboring an expressive biodiversity of plant species with
potential for extracting essential oil (EO) with unique phytochemical profiles, among which
are species of Myrtaceae. Among its various pharmacological activities, the high antioxidant
potential has been highlighted. However, within the Myrtaceae family, there are species in the
Amazon region that reports on their antioxidant potential are scarce in the literature, such as
Calycolpus goetheanus (Mart. ex DC.) O. Berg, popularly known as “goiabinha” or plum of
the beach. It presents itself as a shrub with small edible dark-colored fruits with a sweet and

261
aromatic flavor, native to Brazil, concentrated mainly in the northern region. Within this
context, the objective was to extract the EO from the aerial parts of C. goetheanus with
supercritical CO2 (CO2-SFE) and to evaluate its antioxidant potential. The aerial parts of the
plant (leaves, branches and fruits) were collected in the municipality of Salvaterra, Marajó
Archipelago, Pará, Brazil. The samples were dried by lyophilization (final humidity 13.8%)
and crushed in a knife mill. The extractions were carried out under conditions of temperatures
of 35 and 45 °C and pressures of 150 to 350 bar, obtaining the global yield through the
extraction isotherms. The EOs obtained were evaluated for chemical composition and
antioxidant potential by the DPPH method. From the extraction isotherms, the highest
extraction yields are observed in the operational condition of 45 °C/350 bar (2.11%) and 45
°C/250 bar (1.86%), summing up to four times the yield percentage in the condition of 35
°C/150 bar (0.56%). The obtained EO presented as main components 1,8-cineole, linalool and
(E)-caryophyllene. The results of the antioxidant potential confirm that the EO of C.
goetheanus has good antioxidant capacity. This raw material has promising potential to be
commercially exploited due to its high EO yield and good antioxidant capacity, which has
attracted the attention of different industrial segments, especially the pharmaceutical and food
industries.

Keywords: Calycolpus goetheanus, isoterma de extração, óleo essencial, atividade

antioxidante.

262
 Supercritical extraction of butter from agroindustrial cocoa
residue from the amazon
Jean Maurício L. Pinheiroa,,, Ana Paula S. e Silvaa,, Kelly Roberta P. Pantoja, Rafael Vitti
Motaa,, Gabriela Izane de A. Camposa, Giselle Cristine M. Airesa, Verônica M. S. Bezerraa,
Maria Eduarda F. de Carvalho,b , Raul N. de Carvalho Junior,a,*
a
Federal university of Pará (UFPA), Belém, Brasil.
b
Federal Rural University of the Amazon (UFRA), Belém, Brasil.
*mauricio.edulis@gmail.com

ABSTRACT
Theobroma cacao is a tropical plant native to the Amazon rainforest. Research carried out on
plant species indicates that its fruit known as cocoa, presents incorporated theories of
bioactive compounds in comparison to cocoa tree monocultures. During industrial processes
to obtain finished products such as chocolate and cocoa powder, the husk that surrounds the
fruit grains is separated from the cotyledon (nibs) after the drying or roasting step. Still
during the chocolate production process, the cocoa butter is passed through the high pressure
pressing of the nibs, later the butter remaining from the pressing is removed through the use
of toxic organic solvents that make this butter unfit for human consumption, being then
destined for pharmaceuticals or cosmetics. Studies show that the husk of these grains, despite
being considered acceptable for the industry, has a high content of polyphenols and
methylxanthines such as theobromine and caffeine when compared to nibs, especially in
relation to lipid content. In this sense, supercritical technology is a green and very tolerant
method because it applies low temperatures, presents high selectivity and is free of residues
harmful to health, being able to meet the tolerated requirements and the market. Thus, this
work aimed to characterize the cocoa bean husk (CBH) and evaluate the adjusted butter of
this by-product from the recipe via supercritical technology. Cocoa seeds were collected and
fermented for seven days in wooden boxes, stored for drying by exposure to the sun, and
separating the cotyledon from the CBH. The centesimal characterization of CBH (proteins,

263
lipids, carbohydrates, moisture, ashes and fibers) was carried out, characterization of the
extraction bed in relation to the average diameter of the particles, porosity of the bed,
apparent density and real density. Evaluation of the content of phenolic compounds (TCF) by
the Folin-Ciocalteu method and antioxidant activity (AA) by the DPPH and ABTS methods
in fresh samples. The operational parameters of the process were CO2 flow equal to 3.5
L/min, static period of 20 minutes and dynamic period of 90 minutes, sample mass equal to
10 g, temperatures of 40 and 60ºC and pressures of 200, 300 and 400 bar, with density
ranging from (828.28 to 990.90 kg/m3). The result of the centesimal composition of CBH
showed 21.9% protein, 18.7% lipids, 11.1% carbohydrates, 6.4% moisture, 0.8% ash and
41.3% fiber. The mean particle diameter was 1.056 mm, actual density 1.293 g/cm3, apparent
density 0.299 g/cm3 and bed porosity 0.769. The fresh sample has a TCF of 13.97 mg GAE/g
b.s and AA by the DPPH method equal to 77.72 (μmol.TE/g b.s) and by the ABTS method
equal to 43.46 (μmol.Trolox/g b.s). The mass yields observed ranged from 16 to 37%. The
husk of cocoa beans is considered a residue, however, its butter extracted from the husk of
cocoa beans has great potential due to its high content of bioactive compounds and can be
applied in the production of chocolate or other products for food purposes, the degreased
powder can have numerous applications such as cereal bars.

Keywords: Theobroma cacao, Cocoa bean husk, supercritical extraction.

264
 Extraction of yerba mate (Ilex Paraguariensis) using
near-critical CO2 + hydrated ethanol
Luis I. Granonea, Francisco A. Sánchezb,c, Pablo E. Hegelb,c, Selva Peredab,c,d
a
Departamento de Química y Bioquímica, Facultad de Ciencias Exactas y Naturales, Instituto de
Investigaciones Físicas de Mar del Plata, IFIMAR (CONICET-UNMdP), Deán Funes 3350, 7600 Mar
del Plata, Argentina.
b
Planta Piloto de Ingeniería Química, PLAPIQUI (CONICET-UNS), Camino La Carrindanga km 7,
8000 Bahía Blanca, Argentina.
c
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avenida Alem 1253,
8000 Bahía Blanca, Argentina.
d
Thermodynamics Research Unit, School of Engineering, University of KwaZulu Natal, Howard
College Campus, King George V Avenue, 4041 Durban, South Africa.
*corresponding author email address: *******@*******.

GRAPHICAL ABSTRACT

ABSTRACT
Yerba mate (Ilex paraguariensis) leaves, recognized for their numerous health benefits, are
extensively used in Argentina, Brazil, Paraguay, and Uruguay for the preparation of traditional
infusions. Nevertheless, in line with coffee or tea, there are increasing concerns regarding the
elevated content of caffeine found in yerba mate beverages and its potential adverse effects on
human health [1]. Various well-established industrial decaffeination methods are available,
including organic solvent extraction, water extraction, and supercritical fluid extraction using
carbon dioxide (scCO2, T > 304.1 K, p > 73.7 MPa) [2]. Solvent extraction typically employs
dichloromethane and ethyl acetate, which are solvents subjected to strict emission and health
protection laws. Water extraction is more environmentally friendly, but it exhibits a limited

265
selectivity towards other components, resulting in the co-extraction of aroma precursor
compounds alongside caffeine. In contrast, CO2 is a readily available, non-toxic, and non-
flammable solvent and has a superior selectivity towards caffeine [3,4]. Therefore, extraction
using scCO2 overcomes the limitations associated with both solvent and water extraction
methods. However, it does require the installation and maintenance of costly high-pressure
equipment [2].
A more affordable alternative to scCO2 is liquid CO2 under near-critical conditions (ncCO2).
This extraction method offers most of the benefits and advantages of scCO2 extraction but
requires lower operating temperatures (below 304.1 K) and, more importantly, lower operating
pressures (below 73.7 MPa). As a result, the equipment, maintenance, and operational costs
are significantly reduced [2].
In this work, we studied the decaffeination of yerba mate using a ncCO2 Soxhlet extraction
method at 283 K and 4.5 MPa. Chopped yerba mate leaves are processed in a dried form or
impregnated with hydrated ethanol as a cosolvent. A maximum overall extraction yield of 2.68
wt. % is achieved when the water content in the cosolvent mixture is set at 15.0 wt. %. This
resulted in a 32.8 wt. % reduction in the caffeine content of yerba mate leaves impregnated
with a cosolvent-to-feed ratio of 1.0 g g-1. Interestingly, regardless of the percentage of water
in the cosolvent mixture, less than 1.6 % of the total content of two of the main antioxidant
compounds present in yerba mate leaves, caffeic acid and chlorogenic acid, are coextracted in
the process. The overall extraction yield remains close to the maximum when cosolvents with
a water percentage between 4.4 and 22.5 wt. % are used. In contrast, overall extraction yields
below 1.0 wt. % are obtained when cosolvents with water percentages exceeding 30 wt. % are
used. In order to explain this observation, the thermodynamic phase equilibrium at 283 K and
4.5 MPa for a ternary CO2-ethanol-water mixture is modeled using the group contribution plus
association equation of state (GCA-EoS). The predictions of the model reveal that when the
water percentage in the cosolvent is near that of maximal extraction yield, the CO2-ethanol-
water mixture forms a single liquid phase even at high CO2 contents, which is expected to favor
the extraction process. Under low extraction yield conditions, where the water percentage
exceeds 30 wt. %, the GCA-EoS predicts a phase split into a water-rich and a CO2-rich liquid
phase at low CO2 contents. In this context, it is expected for the CO2-rich liquid phase to exhibit
a rather low solvent power, and for the water-rich phase to act as a diffusion barrier, thus
hindering the extraction.
The use of the ncCO2 Soxhlet extraction method presented in this work serves as proof of
principle for the ncCO2 decaffeination of cosolvent-impregnated yerba mate leaves with
negligible coextraction of antioxidant compounds. Although the high-pressure Soxhlet
extraction is not suitable as a large-scale process, the information it provides holds substantial
potential for the conceptual design of scalable semi-continuous systems.

[1] Ramalakshmi, K. and Raghavan B., Crit. Rev. Food Sci. Nutr. 1999, 39, 441-456.
[2] Pietsch, A., in The craft and science of coffee 2017, Academic Press, 225-243.
[3] Saldaña, M. D. A. et al., J. Agric. Food Chem. 2002, 50, 4820-4826.
[4] Hegel, P. E. et al., J. Supercrit. Fluids 2021, 172, 105200.

Keywords: near-critical CO2 extraction, high-pressure Soxhlet, yerba mate, decaffeination.

266
 Application of green technologies for the valorization of banana
peel from the chips industry: a biorefinery approach
Andrea del Pilar Sánchez-Camargoa,b*, Pedro Henrique Santosb, Laís Benvenuttib, Elkin
Rojas-Orduñaa, Sandra Regina Salvador Ferreirab
a
Grupo de Diseño de Productos y Procesos (GDPP), Department of Food and Chemical Engineering,
Universidad de los Andes, Carrera 1E No. 19 A 40, Bogotá, Colombia.
b
Department of Chemical Engineering and Food Engineering, Federal University of Santa Catarina,
Florianópolis, SC, Brazil
*corresponding author email address: ad.sanchez@uniandes.edu.co

GRAPHICAL ABSTRACT

ABSTRACT

Recently, agro-industrial waste has received special attention as a biomass source for
biorefinery processes to obtain new products, as these waste materials can be good sources of
carbohydrates, proteins, lipids, and nutraceutical compounds [1]. Among fruit waste, banana
peel has been mainly used as an adsorbent for pollutant removal, cattle feed, fertilizer, and
traditional medicinal applications [2-4]. However, this waste can be further utilized as it is a
rich source of polysaccharides such as pectin, lignin, cellulose, and hemicellulose, along with
bioactive compounds that provide potential activities such as antioxidant, antimicrobial, anti-
allergic, antifungal, among others [5, 6]. Considering agro-industrial byproducts as a source of
bioactive compounds, the use of non-conventional techniques sparks significant technological
interest. In this study, we aimed to evaluate and apply non-conventional extraction processes
for valorizing an agro-industrial waste of economic importance to Brazil and Colombia, using
a biorefinery approach. Two samples of banana peel byproduct from the banana chips industry,
one from the Florianópolis region in Santa Catarina (Brazil) and the other from the Granada
region in Meta (Colombia), were utilized. The biomass samples underwent pretreatment and
were subjected to proximate analysis, total phenolic content determination, and pectin analysis.

267
Subsequently, as part of the biorefinery approach, phenolic compounds, oligosaccharides, and
other ethanol-soluble compounds were extracted using pressurized liquid extraction (PLE) as
an alternative to conventional methods. The PLE extraction process (80°C, 100 bar) involved
two stages: firstly, a dynamic pumping of a 70% ethanol solution for 10 minutes, followed by
pumping absolute ethanol for an additional 10 minutes, resulting in a final sample-to-solvent
ratio of 1:6 g/mL. This was done with the intention of subsequently obtaining highly
methoxylated purified pectin through hydrolysis of the PLE-pretreated biomass (using a citric
acid solution at pH 2 and a solvent-biomass ratio of 1:20 g/mL) facilitated by microwave-
assisted extraction (MAE). In this regard, a 32 factorial experimental design with 4 central
points was employed for evaluating the effect of temperature (80-100°C) and hydrolysis time
(5-15 min) on i) the extraction yield of pectin, and ii) galacturonic acid content. The results
showed that for both locations, banana peel had a high content of proteins (8.13-8.67% w.b.),
ash (10.37-15.47% w.b.), carbohydrates (61.43-67.00% w.b.), and pectin (23.91-27.34%). The
PLE extract from the biomass from Brazil exhibited the best chemical profile in terms of total
phenolic content (TPC, 34.35 mg GAE/g extract) and inhibition capacity of DPPH radicals
(178.2 μmol/g extract), ABTS radicals (213.75 μmol/g extract), and FRAP radicals (232.54
μmol/g extract). Regarding microwave-assisted pectin extraction from PLE-pretreated
biomass, similar maximum yields (near to 17.0%) were found for both locations under the
conditions of 100 °C and 15 min of extraction. In contrast to these results, the galacturonic acid
content was higher in the sample from Brazil (37.33%), where lower extraction temperatures
(80°C), regardless of the extraction time, were preferred to obtain better pectin qualities.
Finally, the development of this project provided an alternative application for the reuse of an
agro-industrial waste with economic and environmental importance.

Keywords: Waste valorization, agro-industrial byproducts, non-conventional technologies,

pectin, circular economy

References
[1] Sagar, N. A., Pareek, S., Sharma, S., Yahia, E. M., & Lobo, M. G. (2018). Fruit and
Vegetable Waste: Bioactive Compounds, Their Extraction, and Possible Utilization.
Comprehensive Reviews in Food Science and Food Safety, 17(3), 512–531.
https://doi.org/10.1111/1541-4337.12330
[2] Arunakumara, K., Walpola, B. C., & Yoon, M.-H. (2013). Banana Peel: A Green Solution
for Metal Removal from Contaminated Waters. Korean Journal of Environmental Agriculture,
32(2), 108–116. https://doi.org/10.5338/KJEA.2013.32.2.108
[3] Martínez-Ruano, J. A., Caballero-Galván, A. S., Restrepo-Serna, D. L., & Cardona, C. A.
(2018). Techno-economic and environmental assessment of biogas production from banana
peel (Musa paradisiaca) in a biorefinery concept. Environmental Science and Pollution
Research, 25(36), 35971–35980. https://doi.org/10.1007/s11356-018-1848-y
[4] Pathak, P. D., Mandavgane, S. A., & Kulkarni, B. D. (2016). Valorization of banana peel:
a biorefinery approach. Reviews in Chemical Engineering, 32(6).
https://doi.org/10.1515/revce-2015-0063
[5] Maneerat, N., Tangsuphoom, N., & Nitithamyong, A. (2017). Effect of extraction condition
on properties of pectin from banana peels and its function as fat replacer in salad cream. Journal
of Food Science and Technology, 54(2), 386–397. https://doi.org/10.1007/s13197-016-2475-6
[6] Vu, H. T., Scarlett, C. J., & Vuong, Q. V. (2018). Phenolic compounds within banana peel
and their potential uses: A review. Journal of Functional Foods, 40, 238–248.

268
Theoretical Study of the Fractionation of Glycerol Laurates Using
SC-CO2
Francisco A. Sáncheza,b,*, Natalia S. Cotabarrena,b, Pablo E. Hegela,b, Selva Peredaa,b,c
a
Planta Piloto de Ingeniería Química – PLAPIQUI (UNS-CONICET), Camino “La Carrindanga” Km
7, Bahía Blanca (8000), Argentina
b
Departamento de Ingeniería Química, Universidad Nacional del Sur (UNS), Avda. Alem 1253, Bahía
Blanca (8000), Argentina
c
Thermodynamics Research Unit, School of Engineering, University of KwaZulu Natal, Howard
College Campus, King George V Avenue, Durban 4041, South Africa
*corresponding author email address: fsanchez@plapiqui.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Glycerol monolaurate, known as monolaurin, is a monoglyceride of significant importance
due to its antimicrobial properties and its application as non-ionic surfactants, in cosmetics,
and dietary supplements. An attractive route for its production is the direct esterification of
glycerol with lauric acid. However, this reaction is reversible, resulting in the presence of
reactants along with the three glycerol laurates as reaction products. Among the possible sep-
aration methods, supercritical carbon dioxide (SC-CO 2) extraction stands out for its selectiv-
ity and ease of system removal. Previous studies have demonstrated the selectivity of SC-
CO2 in the separation of mixtures of mono- and diglycerides of carboxylic acids with differ-
ent molecular weights.
The objective of this work is to evaluate the selectivity of SC-CO 2 in separating the key com-
ponents of the reactive mixture, including water, glycerin, lauric acid, and the three glycerol
laurates. Due to limited experimental information on these systems, the use of a predictive
model is required. In this study, we employed the Group Contribution with Association Equa-

269
tion of State (GCA-EOS) to thermodynamically model the system. Previous studies have
shown the effectiveness of this model in predicting phase equilibria of related compounds,
such as triglycerides and biodiesel in the presence of CO 2. Moreover, it has been utilized to
assess the selectivity of CO2 in the separation of the three glycerol acetates.
In this work, we evaluate the effect of temperature, pressure, and solvent ratio for designing a
feasible phase scenario for fractionation of glycerol laurates. The phase equilibrium engineer-
ing allows for the decoupling of the variable effects, which is crucial in designing future ex-
periments regarding the fractionation of reaction products.
Keywords: monolaurin, phase equilibrium engineering, supercritical fractionation, GCA-EOS

270
Kinetics of carotenoids extraction from peels of peach palm fruits
(Bactris gasipaes) by CO2 supercritical
Saulo Edgar G. Souzaa, Eduardo G. O. Menezesb, Renan C. Chistéa, Raul N. C. Juniora,*
a
Federal University of Pará (UFPA), Graduate Program of Food Science and Technology (PPGCTA),
Belém, PA, Brazil
b
Federal Institute of Education, Science and Technology of Rondônia (IFRO), Porto Velho, RO,
Brazil.
*corresponding author: raulncj@ufpa.br

GRAPHICAL ABSTRACT

ABSTRACT
Carotenoids are natural lipophilic pigments widespread in nature that can be used as food
colorant, which is useful for food industries, and play important roles for human health due to
their biological properties, such as provitamin A (few selected structures), antioxidant and
antiinflammatory activities, among others. The development of appropriate green methods to
extract carotenoids from plants is one of the most important steps to obtain high-quality
compounds aiming to use them for both scientific and industrial purposes. Thus, in this study,
we investigated the extraction kinetics of high contents of carotenoids from freeze-dried peels
of peach palm fruits (Bactris gasipaes) by supercritical CO2 extraction. For this purpose,
extraction curves of carotenoids were obtained at 40 °C and 400 bar during 35 min, and these
conditions were chosen due to the high extract yield and total carotenoid contents in
preliminary studies. In addition, other operational parameters for the supercritical CO2
extraction were determined, such as the true density of the extraction bed (1242.9 kg/m3), bed
porosity (0.72), density (990.90 kg/m3) and flow rate of the supercritical fluid (3 L/min). Two
mathematical models, Sovová and Spline, were fitted to the experimental data aiming to predict
the minimal extraction time to obtain high contents of carotenoids from avoiding unnecessary
use of energy, CO2 and other supplies. Both the mathematical models were able to predict the
extraction process with high determination coefficient (R² >0.99), and according to the fitted

271
kinetic curves, two main steps were observed : one characterized by the time for carotenoid
solubilization in the supercritical CO2 (480 s) and the second by the compound diffusivity
during extraction (1380 s). The extraction rate during the first step (kf = 39.31 x 10-5 min-1) was
higher than the second one (ki = 5.32 x 10-5 min-1), suggesting a faster extraction during 480 s,
followed by a gradative increase until 1380 s. By the supercritical CO2 extraction, about 50%
of total carotenoid contents were extracted from the freeze-dried peels of peach palm fruits
after 8 minutes (3.4 mg/g), while 90% was obatined after 23 minutes (12.2 mg/g). Therefore,
in our study, both the mathematical models were important tools to predict efficiently the time
needed for extraction of high yields of carotenoids from peels of peach palm fruits by the
supercritical CO2 extraction. Furthermore, peels of peach palm fruits can be sen as promising
sources of bioactive compounds, which are agroindustrial unexploited wastes in the Amazonia,
and further investigations should be estimulated focusing on the development of more
economical and sustainable production processes.

Keywords: Amazonian fruits, bioactive compounds, natural pigments, mathematical

modelling, functional ingredients.

Financial support: Conselho Nacional de Pesquisa e Desenvolvimento (CNPq, Brazil -

(Projects 408181/2021-4).

272
 First Contact Minimum Miscibility Pressure for Enhanced Oil Recovery:
Tracing Pressure-Composition (P-x) Phase Envelopes and Analyzing
Injection Fluid Effects
Federico E. Benellia,b, Gerardo O. Pisonia,b, Martín Cismondia,b*

a Universidad Nacional de Córdoba (UNC) - Facultad de Ciencias Exactas, Físicas y Naturales. Av.
Vélez Sarsfield 1611, X5016GCA, Ciudad Universitaria, Córdoba, Argentina.
b Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA-
CONICET), Córdoba, Argentina.

*corresponding author email address: martin.cismondi@unc.edu.ar.

GRAPHICAL ABSTRACT

ABSTRACT
Enhanced oil recovery (EOR) encopasses a set of techniques used during reservoir fluid
extraction process, once the reservoir pressure has decreased and is not enough for moving the
fluid to the surface. One important technology corresponds to the injection of a gas stream like
the produced natural gas, nitrogen or CO2 into the reservoir. This injection of gas increases the
pressure of the reservoir, displacing the oil from one or more injector wells to production wells.
During the injection process, a phase equilibria phenomenon occurs, where part of the oil
vaporizes into the injected gas, and part of the injected gas is condensed and diluted into the
oil. This two-phase equilibrium exchange occurs until, eventually, complete miscibility is
reached. Achieving full miscibility between both phases is of high importance since this assures
that the displacement gas is fully used in the process and also no gas can break through. The
minimum pressure at which the injected gas and the oil are fully miscible is known as minimum
miscibility pressure (MMP). EOR by gas displacement is also of high interest from an
environmental point of view when CO2 is used as a displacement gas, since it can be a way to
store the gas generated during different processes in the wells, reducing the amount released
into the atmosphere.

273
This process is usually simulated as a series of discrete steps where on each step, one phase is
in equilibrium with the next step phase, and the other phase is in equilibrium with the previous
step phase. A simpler and more conservative way of estimating the required pressure for a gas
injection EOR operation is considering the first contact minimum miscibility pressure (FC-
MMP), which is the minimum pressure at which the original injection gas and the reservoir
fluid are completely miscible. This value will be higher than the real MMP which develops
through the counter-current multicontact process alluded above. Therefore, the determination
of the FC-MMP already provides a good (and safe) initial guess for the pressure at which the
EOR process should be done.
In this work, we develop and implement an efficient algorithm to determine the FC-MPP by
tracing the phase envelope at fixed temperature and with global composition as a function of a
percentage of injected gas. The implementation is based on the continuation method, an
efficient mathematical method to trace lines for a system of equations where one variable can
be specified.
The algorithm is tested on multiple characterized reservoir fluids from the literature, and can
be used as examples of fluids containing asphaltenes, water and/or CO2. We also study the
effect of different kinds of injection fluids (mostly CO2 and different mixtures of light alkanes),
studying how these different fluids impact the FC-MMP.

Keywords: EOR ; Gas injection ; Reservoir Fluids, Equations of State, Continuation Method.

274
Assembly of supercritical extractor to operate in semi-continuous
mode without CO2 liquefaction
Ana Paula S. Silvaa*, Maria Caroline R. Ferreiraa, Letícia M. M. Siqueiraa, Luã B. Santosa,
Leandro L. T. Ribeiroa, Raul N. Carvalho Jra

a
LABTECS (Supercritical Technology Laboratory), ITEC (Institute of Technology), UFPA
(Federal University of Pará), Rua Augusto Corrêa S/N, 66075-900, Guamá, Belém, Pará,
Brazil

* Corresponding authors: MSc. Ana Paula de Souza e Silva. Phone: +55-91-98805-0945. E-

mail: anapaula-eng@hotmail.com; Dr. Raul Nunes de Carvalho Junior. Phone: +55-91
98742- 0195. E-mail: raulncj@ufpa.br, raulncj@gmail.com.

ABSTRACT
Sustainability and environmental preservation are topics that are increasingly discussed today.
In view of this, society has sought products of natural origin, obtained through processes that
do not cause damage to the environment and do not pose health risks. In view of this, the
supercritical fluid extraction process (SFE – Supercritical Fluid Extraction) has been gaining
ground in the industrial sector as it is considered a clean technology that provides products that
are different from those obtained by conventional processes. The advantages of supercritical
technology are reported in the literature, mainly because of the use of a solvent generally
recognized as safe (GRAS). Many works have been developed with supercritical technology,
however, most are carried out in commercial equipment, which prevents the breakdown of
equipment costs, and restricts the possibilities of customized layouts.

275
Thus, the objective of this work was to assemble a semi-continuous SFE 2x1L supercritical
extraction pilot unit without the CO2 cooling step. The flowsheet of the extraction unit was
developed with the following equipment and instruments: CO2 reservoir, air compressor,
gaseous CO2 pneumatic pump, extraction vessels, separator vessels, blocking, micrometric,
safety, and flow control valves air, backpressures, pressure gauges, flowmeters, flow totalizers
and an automatic temperature control and measurement system. After defining the flowsheet,
the acquisition of equipment and accessories was carried out and the assembly started, after
purchasing all instruments. During the entire unit assembly process, the MASP tool was used
for analysis and problem solving. To validate the extraction unit, a commercial Spe-edTM SFE
equipment (Applied Separations, model 7071) was used, with a 0.1L extraction cell.
Temperature, pressure, bed porosity and raw material batch conditions were kept constant in
both units. Experiments were carried out with Acmella oleracea (jambu), Mauritia flexuosa
(buriti) and Euterpe oleracea (açaí). And the overall yields were compared between the two
extraction units.
The extraction yield of jambu was 13.41% (temperature: 70°C, pressure: 320bar, bed porosity:
0.86), buriti was 42.6% (temperature: 40°C, pressure: 400bar , bed porosity: 0.81) and açaí was
50.8% (temperature: 50°C, pressure: 350bar, bed porosity: 0.88). Considering the yields
obtained in the extraction units, it is initially concluded that the mounted unit performed
similarly to the commercial unit, proving the cost reduction in the assembly of the unit, from
the elimination of the CO2 cooling step.

Keywords: equipment assembly, equipment layout, supercritical extraction, jambu, buriti, açaí.

276
 Supercritical CO2 extracts of ginger (Zingiber officinale R.):
kinetic global yield, polyphenol content, antioxidant activity and
its application as antioxidant in olive oil
Jennifer Meza-Coaquiraa, Fiorella P. Cárdenas-Toroa,*
a
Pontifical Catholic University of Perú, Department of Engineering, Section of Industrial Engineering,
Chemical Engineering Program, Av. Universitaria 1801, Lima, Perú
*corresponding author email address: fcardenas@pucp.pe

GRAPHICAL ABSTRACT

ABSTRACT
Ginger (Zingiber officinale R.) is widely recognized for its therapeutic effects, mainly due to
the presence of gingerol and shogaol, to which antioxidant, anti-inflammatory, and antitumor
properties are attributed, among others [1]. Due to its importance in health, it is currently used
as an additive in the food and cosmetic industry. The extraction of ginger essential oil and
oleoresin using supercritical fluid technology has the advantage over conventional techniques
in relation to the use of low temperatures when carbon dioxide is used as a solvent (from 31°C
and 74 bar). [2]. The separation of carbon dioxide by decompression makes it possible to obtain
an extract without residual solvent. On the other hand, carbon dioxide is considered a Generally
Recognized as Safe (GRAS) solvent, so its use is allowed in the food industry. The selection
and appropriate manipulation of pressure and temperature in the extraction process allows to
selectively separate the essential oil or oleoresin. In the present work, the obtaining of ginger
extracts using supercritical carbon dioxide from powdered lyophilized ginger root was studied.
The experiments were carried out in a commercial 100 mL capacity supercritical extraction
equipment (Supercritical Fluid Inc, USA). The operating conditions used were temperatures of
40, 45 and 50ºC; pressure of 80, 100, 150, 200 and 250 bar; carbon dioxide flow rate of 2 ft3
/h and 3 h of dynamic extraction. The experiments were performed in triplicate. Kinetic
extraction global yield was evaluated in terms of mg extract/g raw material on a dry basis every
10 min. The extracts were analyzed in terms of total polyphenols and antioxidant activity using
the FRAP and DPPH assays. For the studied conditions, essential oil extracts were obtained at
pressure of 80 and 100 bar, while oleoresin extracts were obtained at 150, 200 and 250 bar.

277
The global extraction yields varied in the range of 1.84 ± 0.07 – 16.35 ± 1.10 mg/g d.b. The
polyphenol content varied between 1.48 ± 0.44 and 215.07 ± 0.95 mg GAE/g extract. The
antioxidant capacity was of 82.64 ± 33.23 to 601.91 ± 28.39 mg Trolox/g extract, and IC50
varied between 0.85 – 0.95 mg/mL. The highest extraction recovery was obtained at 50°C and
250 bar, which presented high values of polyphenol content and antioxidant activity. Finally,
the addition of oleoresin extract as an antioxidant in a sample of commercial olive oil showed
an increasing of its polyphenol content and antioxidant capacity. This study suggests the
potential use of ginger oleoresin obtained by supercritical technology as an antioxidant.

Keywords: supercritical extraction, ginger, oleoresin

References:
1. Ozkur, M., Benlier, N., Takan, I., Vasileiou, C., Georgakilas, A. G., Pavlopoulou, A., ... & Saygili, E.
I. (2022). Ginger for Healthy Ageing: A Systematic Review on Current Evidence of Its Antioxidant,
Anti-Inflammatory, and Anticancer Properties. Oxidative Medicine and Cellular Longevity, 2022.
2. Shukla, A., Naik, S. N., Goud, V. V., & Das, C. (2019). Supercritical CO2 extraction and online
fractionation of dry ginger for production of high-quality volatile oil and gingerols enriched
oleoresin. Industrial Crops and Products, 130, 352-362.

278
 Unlocking the potential applications of Ganoderma applanatum
through ultrasound and supercritical fluid extraction: A
presentation of the state of the art
Paulo C. Torres-Mayangaa,*, Juliane Viganob, Larry O. Chañi-Paucarc, Susan R. Tovar-
Barrientosd, Cristian F. Epiquien-Zorrillad, José L. Pasquel-Reáteguie, Luis Condezo-Hoyosa,
Mauricio Rostagnof
a
Innovative Technology, Food and Health Research Group, Departamento de Ingeniería de Alimentos
y Productos Agropecuarios, Facultad de Industrias Alimentarias, Universidad Nacional Agraria La
Molina, Av. La Molina S/N—Lima 12, Lima, Peru
b
Natural science center, Universidade federal de são Carlos, Rodovia Lauri Simões de Barros, km 12 -
SP-189 - Aracaçu, Buri - SP, 18290-000
c
Departamento Académico de Ingeniería Agroindustrial, Universidad Nacional Autónoma Altoandina
de Tarma (UNAAT), Florida-Cochayoc Highway, Huancucro 2092, Zip code: 12651, Junin, Peru
d
Facultad de Industrias Alimentarias, Universidad Nacional Agraria La Molina, Av. La Molina
S/N—Lima 12, Lima, Peru
e
Faculty of Agroindustrial Engineering, Agroindustrial Engineering Department, University National
of San Martín (UNSM), Tarapoto, SM, Peru
f
Multidisciplinary Laboratory of Food and Health (LabMAS), School of Applied Sciences (FCA),
Universidade Estadual de Campinas, São Paulo, Brazil

*Corresponding author email address: paulotorres@lamolina.edu.pe

GRAPHICAL ABSTRACT

ABSTRACT
The fruiting body of Ganoderma applanatum has been used since ancient times due to its
medicinal properties in Asian countries. This species of fungus has great bioactive potential
because it contains secondary metabolites which have pharmacological properties and
immunomodulatory activity of great interest to the pharmaceutical and functional food
industries. Such properties have contributed to it becoming the focus of several research that

279
seek to develop innovative and beneficial products for health. In Peru, Ganoderma spp. grows
wild in the Amazon and has not been the subject of study in relation to the identification of the
species, extraction of bioactive compounds and technological development. Unlike its
counterpart, Ganoderma Lucidum, which has shown potential for the prevention and/or
treatment of diseases, due to its high content of triterpenes, polysaccharides, and
immunomodulatory proteins, generating anti-inflammatory, antioxidant, anti-tumor properties,
among others. For this reason, the valorization of Ganoderma applanatum native to the
Peruvian Amazon must be widely explored, in addition to applying appropriate technologies
that guarantee the quality and safety of the extract with a high bioactive fraction. The objective
of this study is to present the state of the art on the use of the fruiting body of Ganoderma, in
obtaining extracts with antioxidant, antiallergic, anti-inflammatory, antiviral, antitumor
activity, using eco-friendly technologies such as ultrasound and SFE. Pretreatment process will
be performed applying ultrasound as a technique to enhance the supercritical fluid extraction
(SFE) on the fruiting body of Ganoderma applanatum. Ultrasound as a pretreatment will
promote the rupture of the solid matrix through the variation of power and using different
ethanol concentration. The pretreatment will facilitate the access of solvents to the solute
during the SFE that is considered a safe and ecological technique. Supercritical carbon dioxide
(sc-CO2) has proven to be very efficient in the extraction of low polarity compounds, in
addition to being a fast and ideal process for the recovery of biomolecules, minimizing their
degradation due to the low temperatures applied in the process. sc-CO2 is an appropriate
technique for the recovery of triterpenes, steroids, alkaloids, and polysaccharides, which will
be correlated with the applied process conditions. As the main result of this research, we seek
to position the potential of the fungus Ganoderma applanatum, a species native to the Peruvian
Amazon that is currently not exploited. Likewise, it seeks to generate a solid base of knowledge
for future research and applications in the pharmacological and food industries, since it
provides a high benefit in the treatment of different diseases, due to its therapeutic potential.

Keywords: Ganoderma applanatum, extraction bioactive component, potential

pharmacological, Ultrasound pretreatment, supercritical carbon dioxide.

280
 YTEC - Privada

Comparison of Modified HPAM Using Hyperbranched Brush

Macromonomer in Supercritical Carbon Dioxide and Controlled
Polymerization

Ramses S. Meleán Britoa,b, Agustín Iborrac, Joana E. Tasquec, Miriam C. Strumiaa,b, Facundo
Mattea*a,b, Isabel N. Vegac, Juan M. Giussi*c and Juan M. Milanesiob,d
a
Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química
Orgánica. Av. Haya de la Torre y Av. Medina Allende, Córdoba, X5000HUA, Argentina.
b
CONICET, Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada
(IPQA – UNC – CONICET). Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
c
YPF TECNOLOGÍA S.A., Av. Del Petróleo s/n (entre 129 y 143), Berisso, Argentina
d
Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales. Departamento
de Química Industrial y Aplicada. Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
* E-mail: fmattea@unc.edu.ar, juan.m.giussi@ypftecnologia.com

GRAPHICAL ABSTRACT

ABSTRACT
Partially hydrolyzed polyacrylamide (HPAM) has found widespread use in the Oil & Gas
industry in recent years, particularly for Enhanced Oil Recovery (EOR) applications. However,
the viscosity of its aqueous solution is adversely affected by reservoir conditions, such as high
salinity and temperature. Thus, a significant technological challenge is to synthesize HPAM
copolymers capable of maintaining and enhancing their viscosifying properties under harsh
conditions [1,2]. An effective approach for enhancing the viscosity of HPAM involves the

281
incorporation of associative comonomers within the polymer chain. Hyperbranched molecules
like Boltorn H30 with their multiple terminal hydroxyl groups, offer an opportunity for
generating multiple associative interactions and for the customization of hyperbranched
structures to suit specific applications [3]. These modified hyperbranched molecules can be
added to the reactive mixture as co-monomers to obtain modified HPAMs, potentially leading
to copolymers with enhanced resistance to high temperatures and salinity[4,5].
In this study, we partially modified the terminal hydroxyl groups (8 out of 32) of a
hyperbranched molecule (Boltorn H30) using poly(ethylene glycol) methyl ether methacrylate
(PEGMA 500) via atom transfer radical polymerization (ATRP) and then, in order to obtain
polymerizable methacrylates in the macrostructure for the next step, free hydroxyl groups were
derivatized in a considerably smaller proportion with methacrylic anhydride. The aim was to
produce a water-soluble hyperbranched comonomer that could be incorporated into the
modified HPAM polymer chain through radical polymerization.
To obtain the modified HPAMs, two different synthesis pathways were explored: a
polymerization in aqueous solution controlled by iodine, and precipitation polymerization
controlled by iodine in supercritical carbon dioxide. The latter approach facilitates post-
reaction purification and separation steps, commonly associated with polymerizations using
organic solvents or water solutions.
The synthesized polymers were characterized using spectroscopic methods like FTIR and 1H-
NMR to confirm the incorporation of the hyperbranched monomer, and to evaluate the
feasibility and yield of the proposed reactions. Subsequently, the effects of the modification on
the rheological properties of aqueous polymer solutions were also studied, and the polymer
obtained by both synthetic approaches were compared.

Keywords: Bolton H3O, PEGMA, hyperbranched, salinity resistance, ATRP, Iodine controlled
polymerization.

References

[1] D.A.Z. Wever, F. Picchioni, A.A. Broekhuis, Polymers for enhanced oil recovery: A paradigm for
structure–property relationship in aqueous solution, Prog Polym Sci. 36 (2011) 1558–1628.
https://doi.org/10.1016/j.progpolymsci.2011.05.006.
[2] A.J. Scott, L. Romero-Zerón, A. Penlidis, Evaluation of Polymeric Materials for Chemical Enhanced
Oil Recovery, Processes. 8 (2020) 361. https://doi.org/10.3390/pr8030361.
[3] C. Gao, D. Yan, Hyperbranched polymers: from synthesis to applications, Prog Polym Sci. 29 (2004)
183–275. https://doi.org/10.1016/j.progpolymsci.2003.12.002.
[4] L. Shi, C. Liu, M. Chen, Z. Hua, Z. Ye, J. Zhang, Synthesis and evaluation of a hyperbranched
copolymer as viscosity reducer for offshore heavy oil, J Pet Sci Eng. 196 (2021) 108011.
https://doi.org/10.1016/j.petrol.2020.108011.
[5] N. Lai, X. Qin, Z. Ye, Q. Peng, Y. Zhang, Z. Ming, Synthesis and Evaluation of a Water-Soluble
Hyperbranched Polymer as Enhanced Oil Recovery Chemical, J Chem. 2013 (2013) 1–11.
https://doi.org/10.1155/2013/824785.

282
 Supercritical CO2 + co-solvent extraction of essential oil from Mentha
piperita and scale up

Marleny D.A. Saldañaa,*, Yuan Menga

a
Department of Agricultural, Food and Nutritional Science, University of Alberta
116 St & 85 Ave, Edmonton, Alberta, Canada
*marleny.saldana@ualberta.ca

GRAPHICAL ABSTRACT

ABSTRACT

Peppermint essential oil of Mentha piperita is widely used in food and pharmaceutical
products. The objective of this study was to evaluate the use of supercritical carbon dioxide
and co-solvents to extract essential oil from peppermint leaves. To achieve maximum
extraction yield, a set of experiments were designed to evaluate the impact of main processing
parameters such as pressure (100-400 bar), temperature (45-55 C), CO2 flow rate (0.5, 3
mL/min), time (15-180 min) and co-solvent (ethanol, acetone and isopropyl acetate). The
extracts were analyzed by gas chromatography to determine the contents of menthol and
menthone. The highest yield (1.970±0.107%) was achieved at 400 bar and 50C for 150 min

283
with a flow rate of 3 mL/min, however, the highest quality of essential oil was obtained at 110
bar and 50 C for 150 min with a flow rate of 3 mL/min with the highest menthol content
(47.5%) and menthone content (2.3%). All co-solvents increased the yield, but the menthol
content was lower while the menthone content was higher than the pure SC-CO2. The 3-fold,
5-fold and 10-fold scale-up extraction achieved yields of 0.94%, 0.79%, and 0.77%,
respectively, at 110 bar and 50°C. In addition, the scale-up process led to slightly higher, but
not significant, menthol contents of 49.8%, 48.2%, and 47.4%, and slightly lower, but not
significant, menthone contents of 1.9%, 2.0%, and 2.1% in the extracts. Peppermint essential
oil has demonstrated significant potential as an effective antioxidant, with the highest
antioxidant activity observed at a concentration of 500 mg/mL with 77.9% of DPPH
scavenging activity and 82.3 µM ascorbic acid equivalent.

Keywords: Supercritical carbon dioxide extraction; co-solvent; scale up; peppermint essential
oil; menthol; menthone.

284
 Exploring the Physicochemical and Rheological Properties of
Acrylic Acid - co - 2-Acrylamido-2-methylpropane Sulfonic Acid
Copolymers Synthesized in Supercritical Carbon Dioxide
Ramses S. Meleán Britoa,b, Facundo Matteaa,b, Juan Padróc, Miriam C. Strumiaa,b, Séverine
Camyd, Mathias Destarace and Juan M. Milanesio*b,f
a
Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química
Orgánica. Av. Haya de la Torre y Av. Medina Allende, Córdoba, X5000HUA, Argentina.
b
CONICET, Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada
(IPQA – UNC – CONICET). Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
c
YPF TECNOLOGÍA S.A., Av. del Petróleo s/n (entre 129 y 143), Berisso, Argentina
d
Laboratoire de Génie Chimique, CNRS/UT3/INP, Université de Toulouse, 31030 Toulouse, France
e
Laboratoire des IMRCP, Université Toulouse 3 Paul Sabatier, CNRS UMR 5623, 118 route de
Narbonne, 31062 Toulouse, France
f
Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales. Departamento
de Química Industrial y Aplicada. Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
*
E-mail: juan.milanesio@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Acrylic acid (AAc) and 2-acrylamide-2-methylpropane sulfonic acid (AMPS) are vinylic
monomers that can be polymerized using solution or precipitation techniques, utilizing either
water or organic solvents. Depending on whether water or an organic solvent is used, suitable
thermal initiators like azo compounds, peroxides, and persulfates, can be employed. After the
reaction, precipitation and drying processes must be carried out to obtain a solvent-free

285
polymer [1]. In precipitation polymerization, azo compounds are the preferred thermal
initiators. At the beginning of the reaction, a homogeneous mixture of solvent, monomers, and
initiators is prepared. As the reaction proceeds, the polymer precipitates, resulting in a polymer-
rich phase, that can be liquid or solid. An energy-intensive drying process is required at the end
of the reaction.
As an alternative approach, compressed solvents like carbon dioxide (CO2) can be used as
solvents for precipitation polymerization [1]. Due to its adjustable density and solvent power,
supercritical CO2 can effectively solubilize AAc and other vinylic monomers. An advantage of
using CO2 lies in its capacity to yield high-purity end products via a straightforward
depressurization of the system, avoiding precipitation or drying processes. Additionally, CO2
can be recycled by recompression after the reaction, thereby minimizing the presence of toxic
organic solvents that could have an impact in human health [2]. CO2 is an inert solvent, showing
no radical transfer reactions, making it a suitable choice for radical precipitation
polymerization. Previous studies have proved that CO2 can be used in the synthesis of
poly(acrylic acid) yielding higher average molecular weights compared to polymers
synthesized in water or organic solvents [3].
This study focuses on the precipitation copolymerization of AAc with AMPS in supercritical
CO2, with a reactive mixture ranging from 10 to 50 wt.% AMPS. The incorporation of AMPS
into the polymer chain was investigated by means of Fourier Transform Infrared (FTIR)
Spectroscopy and Proton Nuclear Magnetic Resonance (1H-NMR), the average molecular
weight distributions and viscosity of the resulting polymeric solutions in water were examined
by using Size Exclusion Chromatography (SEC) and rheology measurements, respectively.
Additionally, the effect of variations of pH and salinity over the viscosity of the polymer
solution was analyzed. The results showed the incorporation of AMPS and significant viscosity
increases in water solution, particularly with AMPS contents up to 30 wt.%. Furthermore, the
polymers displayed resistance to solution salinity as evidenced by the slight variation in the
viscosity with increasing sodium chloride (NaCl), and calcium chloride (CaCl2) salt
concentration.
Keywords: Carbon dioxide, supercritical, rheology, 2-acrylamide-2-methylpropane sulfonic
acid, salinity resistance, precipitation polymerization.

References
[1] S. Nazaripour, M. Rafizadeh, H. Bouhendi, Percipitation copolymerization of acrylamide and acrylic
acid: Determination of reactivity ratio by various methods, E-Polymers. 12 (2012) 1–16.
https://doi.org/10.1515/epoly.2012.12.1.181.
[2] T. Liu, P. Garner, J.M. DeSimone, G.W. Roberts, G.D. Bothun, Particle Formation in Precipitation
Polymerization: Continuous Precipitation Polymerization of Acrylic Acid in Supercritical Carbon
Dioxide, Macromolecules. 39 (2006) 6489–6494. https://doi.org/10.1021/ma061260p.
[3] J.N. Ollagnier, T. Tassaing, S. Harrisson, M. Destarac, Application of online infrared spectroscopy to
study the kinetics of precipitation polymerization of acrylic acid in supercritical carbon dioxide, React
Chem Eng. 1 (2016) 372–378. https://doi.org/10.1039/c6re00022c.

286
 Exploring the potential of Supercritical Carbon Dioxide for
Eugenol impregnation in 3D printed Polylactic Acid Structures
Janet Chinellatoa,b, Marcelo R. Romeroa,b, Facundo Matteaa,b*
a
Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química
Orgánica, Av. Haya de la Torre y Av. Medina Allende, Córdoba, Argentina
b
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA-
CONICET), Edificio de Ciencias II. Haya de la Torre y Medina Allende, Córdoba, Argentina.
*fmattea@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
The development of antimicrobial polymeric systems holds significant promise in the battle
against the spread of diseases, thereby enhancing health conditions in the field of medical
science and the food industry. In addition, the creation of degradable materials plays a crucial
role in addressing the global plastic pollution.
One of the synthetic polymers showing great potential in replacing conventional plastics is
polylactic acid (PLA). PLA, a versatile polyester, is recognized for its biocompatibility,
biodegradability, and bioresorbability, finding applications in various fields such as medical
science, engineering devices, and food packaging (Singhvi et al., 2019; Malek et al., 2021).
Notably, PLA possesses favorable mechanical properties for 3D printing, making it a viable
alternative for constructing prostheses (Wu et al., 2019). Furthermore, the ability of PLA to be
reabsorbed by the body's biochemical systems enhances its value as a biopolymer for tissue
engineering applications. This characteristic eliminates the need for additional surgeries in

287
patients with implants or cell regeneration devices, presenting promising prospects in the field
(Singhvi et al., 2019; Malek et al., 2021).
The functionalization of PLA by incorporating antimicrobial active compounds is an area that
is experiencing continuous growth and exploration, especially with substances from natural
sources. Among the promising antimicrobial compounds, eugenol stands out as a noteworthy
candidate. Eugenol is a key volatile component found in clove essential oil, renowned for its
antimicrobial properties. However, its benefits extend beyond just antimicrobial activity, as it
also exhibits anti-inflammatory, analgesic, antioxidant, and even anticancer properties
(Kamatou et al., 2012).
Currently, there is limited research available on systems that utilize both PLA and eugenol (Li
et al., 2021; Huang et al., 2022). To the best of our knowledge, the use of supercritical carbon
dioxide (ScCO2) assisted impregnation for incorporating eugenol into PLA has not been
reported in the literature. Supercritical carbon dioxide (ScCO2) has been recognized as a clean
alternative to conventional impregnation techniques. Its inert nature ensures that any residues
left on the material are harmless, making it an environmentally friendly option Additionally,
ScCO2 is cost-effective, non-flammable, and easy to recycle or dispose of (Mosquera et al.,
2019).
Taking all these into account, the aim of the present work was to study the ScCO2 impregnation
of eugenol into 3D structures of commercial PLA. The 3D structures were printed using a
Fused deposition modeling (FDM). Impregnation runs were carried out in a lab-scale high
pressure system, evaluating the effect of different operation variables on impregnation loading,
morphology of the printed structures, and active compound distributions within printed
structures. The analysis of the 3D structure, before and after the ScCO2 impregnation, was
carried out by reflectance confocal optical microscopy technique and the distribution of
eugenol was compared by means FTIR measurements.

Keywords: Polylactic acid, eugenol, supercritical CO2 impregnation

References

Singhvi, M.S.; Zinjarde, S.S.; Gokhale, D.V. (2019). Polylactic acid: synthesis and biomedical applications,
Journal of Applied Microbiology 6 (127), 1612-1626.
Malek, N.S.A.; Faizuman, M.; Khusaimi, Z.; Bonnia, N. N.; Rusop, M.; Asli, N.A. (2021). Preparation and
Characterization of Biodegradable Polylactic Acid (PLA) Film for Food Packaging Application: A
Review, Journal of Physics: Conference Series, (1892), 1-7.
Wu, D.; Spanou, A.; Diez, A.; Persson, C. (2020). 3D-printed PLA/HA composite structures as synthetic
trabecular bone: A feasibility study using fused deposition modeling, Journal of the Mechanical Behavior
of Biomedical Materials 103, 1-10.
Li, M.; Yu, H.; Xie, Y.; Guo, Y.; Cheng, Y.; Quian, H.; Yao, W. (2021). Fabrication of eugenol loaded gelatin
nanofibers by electrospinning technique as active packaging material, LWT- Foof Sciencie and
Technology 139, 110800.
Kamatou, G.; Vermaak, I.; Viljoen, A. (2012). Eugenol—From the Remote Maluku Islands to the International
Market Place: A Review of a Remarkable and Versatile Molecule, Molecules 17 (6), 6953-6981.
Huang, X.; Ge, X.; Zhou, L.; Wang, Y. (2022). Eugenol embedded zein and poly(lactic acid) film as active food
packaging: Formation, characterization, and antimicrobial effects, Food Chemistry 384, 132482.
Mosquera, J.E.; Goñi, M.L.; Martini, R.E.; Gañán, N.A. (2019). Supercritical carbon dioxide assisted
impregnation of eugenol into polyamide fibers for application as a dental floss, Journal of CO2 Utilization
32, 259-268.

288
 Delipidation of Porcine Femur Bone and Porcine Pericardial
Membrane Using Supercritical CO2 Extraction

Julian Reynosoa, Nicolas Gañan b,c, Alexis Velez b,c, Mariano Muraturea and Juan M.
Milanesio*b,c
a
InBiomed S.A Ingeniería Biomédica
b
CONICET, Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada
(IPQA – UNC – CONICET). Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
c
Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales. Departamento
de Química Industrial y Aplicada. Av. Velez Sarsfield 1611, Córdoba, X5016GCA, Argentina.
*
E-mail: juan.milanesio@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
It is well-established that to accelerate biological mechanisms for periodontal bone healing,
guided tissue regeneration (GTR) and guided bone regeneration (GBR) techniques are
employed. The common dental practice today involves the use of porcine pericardial
membranes, composed of native collagen and combined with bone graft materials, in both GTR
and GBR procedures. Porcine pericardial membranes are collagen-rich and exhibit rapid
resorption. They are easily handled due to their high elasticity, facilitating their adherence to
the defect.
A critical step in the utilization of porcine pericardial membrane and femur bone for
assimilation in dental applications is delipidation, as it ensures their absorption by the
periodontal tissue. The residual lipids in bone act as a barrier to cell attachment, adversely
altering its biocompatibility and osseointegration. Furthermore, it induces adverse reactions

289
which can increase bone resorption and encapsulating fibrosis [1]. Nevertheless, these
disadvantages can be resolved by decellularization of bone materials using supercritical carbon
dioxide (SCCO2) extraction. CO2 is the extracting solvent to remove the fats, cells, and non‐
collagenous proteins from the animal tissues. CO2 is a natural occurring solvent, safe, non‐
toxic, noncorrosive, non‐flammable, easily accessible, and cost effective. It presents mild
critical coordinates, such as 7.38 MPa and 31°C, which can be relatively easily accessible [2].
The supercritical CO2 extraction tests were carried out in a high-pressure batch system, which
comprised a 100 cm3 stainless steel cell with internal stirring and temperature control. The
extraction was conducted in two stages. The ethanol-wet bone or membrane sample was placed
inside the extractor and CO2 was subsequently injected using a syringe pump until the desired
pressure was reached. After stabilizing the pressure and temperature conditions (40°C and 200
bar), the system was left in static mode for 1 hour with continuous stirring. After static time,
the cell was depressurized, the sample was removed, and its weight was measured to determine
mass loss. The extractor cell was cleaned and dryed and the same sample was reloaded. The
cycle was repeated for an additional hour. The results were compared against Soxhlet
extraction to evaluate delipidation efficiency. The goal of this work was to assess the
supercritical CO2 lipid extraction from porcine femoral head bone and from porcine pericardial
membrane and characterize the fatty acid profile of the extracted lipids.
The results show that supercritical CO2 extraction facilitated the delipidation of bone and
membrane samples through a two-stage batch extraction scheme, achieving a 25% extraction
yield. This suggests that a semi-continuous operation with a constant flow of supercritical CO2
for delipidation could be more efficient. The extraction technique did not exhibit diffusion
issues, as the lipid extraction percentages remained consistent across various sizes of porcine
bone and membrane.
Keywords: Supercritical, rheology, associative polymer, ternary system, polyacrylic acid.

References

[1] Chen, Y.-W., Hsieh, D.-J., Periasamy, S., Yen, K.-C., Wang, H.-C. and Chien, H.-H. (2021),
Development of a decellularized porcine bone graft by supercritical carbon dioxide extraction
technology for bone regeneration. J Tissue Eng Regen Med, 15: 401-414.
https://doi.org/10.1002/term.3181
[2] J. Fages, A. Marty, C. Delga, J-S Condoret, D. Combes, P. Frayssinet. (1994), Use of
supercritical CO2 for bone delipidation. Biomaterials 15: 650-656. https://doi.org/10.1016/0142-
9612(94)90162-7

290
 Supercritical CO2 pretreatment and subcritical water extraction
of sulfated flavonoids from Flaveria bidentis leaves
Paulina Fallettia,b,*, María F. Barrera Vázquezb, Pedro H. Santosc,
Luiz G. Gonçalves Rodriguesc, Marcelo Lanzac, Raquel E. Martinia,b, Laura R. Cominid,e.
a
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA –
CONICET), Av. Vélez Sársfield 1611, Córdoba, Argentina.
b
Facultad de Ciencias Exactas, Físicas y Naturales (FCEFyN), Universidad Nacional de Córdoba
(UNC), Av. Vélez Sársfield 1611, Córdoba, Argentina.
c
Laboratório de Termodinâmica e Tecnologia Supercrítica (LATESC), Universidade Federal de Santa
Catarina (UFSC), Florianópolis, Brasil.
d
Centro de Excelencia en Productos y Procesos Córdoba (CEPROCOR), Santa María de Punilla,
Argentina.
e
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
*corresponding author email address: pfalletti@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Flaveria bidentis is an annual weed native to South America and has spread rapidly, posing
serious threats to crop and native biodiversity in different countries. This species is capable of
synthesizing two quercetin derivatives with the highest degree of sulfation known: quercetin
3,7,3',4'-tetrasulfate (QTS) and quercetin 3-acetyl-7,3',4'-trisulfate (ATS), which present
important antithrombotic pharmacological activities and inhibitors of the aldose reductase
enzyme. These sulfated flavonoids (SF) are not commercially available and they are obtained
exclusively from the plant kingdom. Thus, large volume of biomass generated by F. bidentis
during its weeding can be exploited in order to obtain these bioactive compounds. However,
they are generally extracted through conventional methods that present low yields and
extraction efficiencies and for this reason, alternative extraction methods should be explored.

291
In this sense, the extraction of natural compounds with subcritical water (SWE) implies the
use of water as extraction solvent, at temperature between 100-374°C and pressure high
enough to maintain it in a liquid state. SWE is considered a green technology because it is
ecological, economical and safe. On the other hand, supercritical CO2 (SC-CO2) has been
used to carry out sample treatments prior to the extraction of the active principles, to weaken
the cell walls of the plant matrix and increase its recovery.
Thus, in this work we propose the design, development and optimization of the pretreatment
of F. bidentis leaves with SC-CO2 (P-SC-CO2), and its subsequent SWE extraction, with the
aim to increases the extraction yield and efficiency of total SF (QTS+ATS), in comparison
with the extractive method in the absence of pretreatment (SWE). For this purpose, the SWE
was performed under optimal conditions previously determined (Falletti et al., 2023b).
During each P-SC-CO2 assay, 3 g of leaves (420 - < 1190 µm) were pretreated at 50°C. The
CO2 was removed by depressurization and the pretreated plant material was reserved for later
extraction. To optimize and evaluate the effects of time (15 and 30 min), pressure (15 and 30
MPa) and depressurization speed (0.2 and 2 Kg CO2/h), on the SWE performance of total SF
(mg of total SF/ 100 g of leaves), a Response Surface Methodology (RSM) was applied based
on a 23 factorial experimental design of two replicates. Data analysis was performed using
ANOVA, significance with p value < 0.05, suitability of the model through the coefficient of
determination R2, and maximization of the desirability function to determine the optimal
conditions. All pretreated samples were subjected to SWE under optimal conditions (2.86
ml/min, 101.2 °C, 420 -< 1190 µm, 100 bar, 35 min, continuous flow) (Falletti et al., 2023b).
The extracts were purified in total SF and finally quantified by high performance liquid
chromatography (HPLC). Additionally, the effect of P-SC-CO2 on plant material morphology
was studied by scanning electron microscopy (SEM) (10 kV, 500X and 1000X, gold coating).
In the SEM images it can be seen that the leaves subjected to P-SC-CO2 show significant
fragmentation due to the high speed of depressurization used. Regarding the RSM, the
ANOVA showed a first-order polynomial equation, with R2=0.99 and a statistically
significant (p<0.05) and positive relationship between the total SF extraction yield and the
conditions of: depressurization speed (most influential factor), pretreatment time, and
interaction between both factors, and the interaction between time and pressure (less
influential). Finally, the optimal conditions of P-SC-CO2 were the highest levels tested for
each of the three variables under study (30 min, 30 MPa and 2 kg CO2/h). A good
concordance was obtained between the theoretical and the experimental value (n=3). The
yield obtained under optimal conditions of the combined method (P-SC-CO2+SWE) was
72.45 mg total SF/100 g leaves. The yield obtained under optimal conditions of the combined
method was 1.8 times higher than the optimized extractive method in the absence of
pretreatment. However, the efficiency of the process (yield/h) was practically the same, being
1.3 times higher than the optimized conventional method (Falletti et al., 2023a).

Keywords: Supercritical CO2 pretreatment, Subcritical water extraction, Sulfated flavonoids,

Optimization, Response surface methodology.

a
Falletti P., Barrera Vázquez M., Cabrera J., Martini R., Comini L. DOI: 10.1007/s12649-
023-02086-8. JCR IF: 3,449 - 2022.
b
Falletti P., Barrera Vázquez M., Gonçalves Rodrigues L., Santos P., Lanza M., Cabrera J.,
Martini R., Comini L. DOI: 10.1016/j.supflu.2023.105958. JCR IF: 4,514 - 2023.

292
 Esterification of oleic acid using subcritical isoamyl alcohol
Maricel Santosa,b, Carolina Rodriguez Ruiza,b, Juan Milanesioa,b, Ivana Magarioa,b, Alexis
Veleza,b*

a Universidad Nacional de Córdoba (UNC) - Facultad de Ciencias Exactas, Físicas y Naturales. Av.
Vélez Sarsfield 1611, X5016GCA, Ciudad Universitaria, Córdoba, Argentina.
b Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA-
CONICET), Córdoba, Argentina.
*corresponding author: avelez@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
In the current environmental context, two objectives are of particular importance. Firstly, the
quest for bioproducts to replace non-biodegradable and/or non-renewable source-derived
products. Secondly, the valorization of by-products to maximize resource yields, results in
improved profits, making such processes more attractive for investment and the replacement
of unsustainable methods.
Fusel oil, a by-product of the bioethanol fermentation process, could be utilized to produce
emulsifier substances. Fusel oil is a mixture of water and various alcohols, including ethanol,
butanol, and isoamyl alcohol, being the last in the major proportion of the blend. Currently,
this by-product is primarily used for energy production in bioethanol plants. However, the
components of fusel oil can also be transformed into more valuable compounds. The
esterification with carboxylic acids is one of the possibilities. Long-chain carboxylic acids such
as oleic acid can be obtained from vegetable oils on a large scale. The resulting products can
be used as bio-emulsifiers or bio-lubricants in the cosmetics, pharmaceutical, and food

293
industries, among others. Furthermore, oleochemical esters have very low or negligible toxicity
and are generally biodegradable.
The production of esters using sub or supercritical alcohols has gained relevance in recent
years. Firstly, it does not require a catalyst, which eliminates the need for neutralization and
separation steps typically found in conventional esterification processes catalyzed by strong
acids or bases. Secondly, the high temperatures involved in the process often significantly
enhance the conversion of reactants and the reaction rate, resulting in the products being
obtained in a shorter time. In the case of reactions involving biphasic mixtures at moderate
temperatures, the use of elevated temperatures promotes the mutual solubility of the reagents
and reduces limitations due to mass transfer phenomena.
In the present study, the esterification of technical-grade oleic acid (77% purity) with isoamyl
alcohol was carried out under subcritical conditions. The experiments were conducted in a
continuous tubular reactor of 16,4 mL at a fixed pressure of 100 bar. The effect of temperature
on the reaction rate and the final conversion of the acid was investigated over the range of
220°C to 300°C, using a fixed molar ratio of isoamyl alcohol to oleic acid of 3:1. Similarly,
the influence of the molar ratio of isoamyl alcohol to oleic acid on the same responses was
analyzed within the range of 1:3 to 3:1. Mass flow rates used in the experiments ranged from
0.4 to 5.6 g/min. Acid conversion over the mass flow rates was monitored through the acidity
index of the collected product samples. The presence of esters was confirmed through gas
chromatography with a Flame Ionization Detector (FID).
The results showed that temperature had a positive effect on the conversion of oleic acid, with
higher temperatures resulting in higher conversion rates. The conversion values of oleic acid
achieved ranged from 70% to 85% for the lowest mass flow rates. In the experiments conducted
at temperatures of 300°C and 260°C, the final conversion rates were nearly identical.
With regard to the initial molar ratio of isoamyl alcohol to oleic acid, an excess of either of the
two reagents resulted in an increase in the conversion rate. At the lowest tested mass flow rate,
a 1:1 molar ratio achieved a 64% conversion. Meanwhile, for ratios of 3:1 and 1:3, the
conversion values of the limiting reagent were similar, at 85% and 82%, respectively.
The obtained results were promising and serve as the starting point for future studies that will
delve into aspects such as reaction kinetics and the utilization of fusel oil as a reactant. The
presence of water in fusel oil will potentially modify the expected conversion percentages, as
water is a product of the reversible esterification reaction studied in this work.

Keywords: fusel oil, esterification, bio-emulsifiers, bio-lubricants

294
 Validation of the RK-PR equation of state for the calculation of
volumetric properties for hydroethanolic mixtures at subcritical
conditions
Andrea C. Rodríguez-Ruiza,b, Gerardo O. Pisonia,b,*, Alexis R. Veleza,b
a
Universidad Nacional de Córdoba (UNC) - Facultad de Ciencias Exactas, Físicas y Naturales. Av.
Vélez Sarsfield 1611, X5016GCA, Ciudad Universitaria, Córdoba, Argentina.
b
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA-
CONICET), Córdoba, Argentina.
*pisoni.gerardo@gmail.com

GRAPHICAL ABSTRACT

ABSTRACT

The family of equations of state (EoS) stemming from the Van der Waals EoS has been
employed for decades to model phase equilibria in systems of varying natures. This family is
characterized by being cubic with respect to volume and featuring attractive and repulsive
parameters, both of which depend on the critical constants (Tc and Pc) of the pure component.
When applying the equation of state to mixtures, a mixing rule for calculating these parameters
should be used. The Redlich-Kwong, Soave-Redlich-Kwong, or Peng-Robinson EoS can be
mentioned as the most widely used within this family, and although these equations yield
satisfactory results in reproducing equilibrium data, they encounter difficulties in accurately
representing volumetric properties of both pure components and mixtures. This problem
becomes more pronounced when modeling highly asymmetric mixtures. The RK-PR EoS
belongs to the Van der Waals family of equations of state and incorporates a third parameter
to adjust the volumetric properties of pure compounds, thus enhancing the accuracy in
reproducing these properties. It's important to note that asymmetric mixtures composed of
compounds capable of forming hydrogen bonds (associations) present an additional challenge.
Typically, these types of mixtures are modeled using equations of state designed to account for

295
such associations, but these equations tend to be more complex than the ones described earlier,
making their implementation a more laborious process. In this context, the objective of this
study is to employ the RK-PR EoS coupled with cubic mixing rules to model experimental data
for an associative system. The primary focus lies in accurately calculating volumetric
properties of the mixture, including density and molar volume. For model validation,
experimental P-T-x data of bubble points of the water + ethanol system under isoplethic and
subcritical conditions were used. These bubble points were obtained using an isochoric method,
where the phase transition points from liquid + vapor to a single-phase system are detected by
the change in slope of the isochore on a pressure vs. temperature graph. Two different values
of ethanol molar fractions were used for the bubble point measurements: 0.4 and 0.6. The
experimental setup used for the isochoric experiments consisted of a stainless-steel constant-
volume cell with a capacity of 5.48 ml, sealed at both ends. The experimental transition points
obtained provide the volumetric data, which are subsequently reproduced using the RK-PR
EoS for modeling equilibrium data. The isochoric method has proven to be a simple and
effective technique for detecting phase transition points in fluid systems. Furthermore, the RK-
PR EoS has demonstrated a strong capability for thermodynamic modeling of hydroethanolic
mixtures. It can be utilized to predict phase envelopes of the water + ethanol system under
subcritical conditions and to predict the volumetric properties of the system in equilibrium with
acceptable accuracy.

Keywords: RK-PR equation of state, hydroethanolic mixtures, volumetric properties,

subcritical conditions

296
 Supercritical CO2 sorption and diffusion in polylactic acid films:
experimental study and modeling
María L. Goñi, José A. Scilipoti, Nicolás A. Gañán*

IPQA (UNC – CONICET), Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina.

ICTA, FCEFyN, Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina.
*corresponding author email address: nicolas.ganan@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Polylactic acid (PLA) appears as a biodegradable and biocompatible alternative to petroleum-
based polymeric materials with several applications in food, pharmaceutical, and biomedical
industries. In this sense, the incorporation of active ingredients such as drugs or natural
compounds using supercritical CO2 (scCO2)-assisted impregnation technology results in an
interesting strategy for developing sustainable active materials. Nevertheless, the design and
modeling of impregnation processing require a deep investigation of the interaction of the
different components (solute-polymer-scCO2).
In the framework of a project concerning the development of PLA-based active materials, this
contribution presents an experimental study of scCO2 sorption into PLA and the subsequent
desorption in ambient conditions, aiming to obtain relevant equilibrium and kinetics data. For
this purpose, commercial PLA films (Converflex, Argentina) and extra dry CO2 (Linde,
Argentina) were used. The sorption experiments were performed in a 50 ml high-pressure
vessel in batch mode [1] at different pressure (5 – 15 MPa) and temperature (45 – 60 °C)
conditions. Immediately after depressurization, the desorption kinetics of CO2 from the films
at ambient conditions was assessed by registering the weight loss of the CO2-swollen films
using a precision digital balance (± 0.0001 g). Desorption curves were extrapolated in order to
obtain equilibrium sorption data.
PLA showed a high scCO2 sorption capacity, obtaining values up to 17 wt% when low
temperature and high-pressure conditions were used (45°C and 15 MPa), which indicates that

297
sorption is extremely related to scCO2 density. Experimental sorption equilibrium data were
modeled with the Sanchez-Lacombe equation of state (SL-EOS) [2], using characteristic
properties for PLA and CO2 taken from the literature after a comparative analysis of reported
values, and fitting the binary interaction parameter kij to minimize the CO2 chemical potential
difference in both phases.
A Fickian mass transfer model for non-steady diffusion in thin slabs was fitted to the
experimental desorption kinetics data, in order to estimate scCO2 apparent diffusivity on PLA
(Da) at ambient conditions. A good correlation with the initial amount of CO2 in the polymer
was obtained, observing an exponential dependency of Da with the sorption values.
The high scCO2 sorption capacity of PLA suggests an important swelling degree of this
polymer, which was also observed and reported by other authors [3]. In a ternary system,
swelling allows a faster solute diffusion through the polymer matrix which favors the
impregnation process. In this sense, PLA can be considered an attractive material for this kind
of process. Finally, the SL-EOS model was successfully fitted for the [scCO2-PLA] binary
system, as a previous step for the predictive modeling of different [solute-PLA-scCO2] ternary
systems.

Keywords: PLA, CO2 sorption, SL-EOS, Fickian diffusion

References:
[1] P. Miranda et al., J CO2 Utiliz, 2022, 61: 102029.
[2] I. Sanchez and R. Lacombe, Macromolecules, 1978, 11: 1145-1156.
[3] R. Pini et al., J Polym Sci, 2008, 46: 483-496.

298
 Preparation of cellulose aerogels using supercritical drying:
Influence of process variables on aerogel porosity
Noelia D. Machadoa,b*, Valentina Piginoa,b, Josefina Chiavassaa,b, María L. Goñi,a,b, Nicolás
A. Gañána,b*
a
Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada (IPQA-UNC-
CONICET), Av. Vélez Sarsfield 1611, X5016GCA, Córdoba, Argentina.
b
Universidad Nacional de Córdoba, Facultad de Ciencias Exactas, Físicas y Naturales, Instituto de
Ciencia y Tecnología de los Alimentos (ICTA-FCEFyN-UNC), Av. Vélez Sarsfield 1611,
X5016GCA, Córdoba, Argentina.
* nmachado@unc.edu.ar, nicolas.ganan@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Aerogels (AE) are lightweight solid materials with an open interconnected structure, high
porosity, and specific surface areas, which are drawing increasing attention in many scientific
fields such as biomedical, energy storage, and others. They can be prepared using different
technologies and a wide range of polymers, therefore, their chemistry, pore size, and surface
area can be tailored. Cellulose is a biopolymer of great interest due to its renewable character,
commercial availability, biocompatibility, and biodegradability, emerging as a promising
source to obtain biobased and biocompatible AE. Cellulose AE can be prepared in 3 steps:
dissolving/dispersing cellulose, forming a hydrogel by the sol-gel process, and gel drying. The
latter step can be performed at ambient evaporation, solvent sublimation by lyophilization, or
under supercritical conditions. The main goal is to avoid pores collapse due to capillary
pressure, retaining the 3D porous structure. In this sense, drying by supercritical CO2 (scCO2)
is considered an appropriate alternative, being a safe and “green” method suitable for most

299
kinds of gels. Before scCO2 drying, the liquid phase of the gel is replaced by organic solvents
miscible with CO2 such as ethanol to form alcogels. This process avoids the liquid-gas surface
tension and liquid-solid adhesive forces preventing the gel collapse. Furthermore, during
scCO2 drying, single-phase fluid conditions can be achieved by setting the process variables,
avoiding pore collapse due to surface tension forces.1
In this work, AE based on cellulose microcrystalline (MCC) were prepared using static scCO2
drying. The effect of different process variables on the final AE structure was studied aiming
to contribute to the knowledge of the basis of scCO2 drying and its optimization.
The MCC hydrogels were obtained after thermal gelation of alkaline solutions of 7 wt.% using
two aqueous solvent mixtures, NaOH:urea 12/7 wt.% and NaOH:urea:thiourea 8:8:6.5 wt.%
according to the literature.2 MCC alcogels were obtained after a gradual solvent exchange using
10-100 % v/v ethanol solutions. Finally, they were dried for 2 h using scCO2 under different
conditions in batch mode. Four process variables were studied, CO2 density (689 kg m-3 at 12
MPa/40°C, and 604 kg m-3 at 15 MPa/60°C), depressurization rate (0.4 and 1.5 MPa min-1),
drying cycles (1 cycle of 2 h and 2 cycles of 1 h each), and composition (AU= MCC-
NaOH/urea and AUT= MCC-NaOH/urea/thiourea). The textural properties (SBET= specific
surface area, and DP= pore diameter), morphology, crystallinity, and typical functional groups
were determined by N2 adsorption/desorption isotherms, SEM, XRD, and FT-IR, respectively.
Among all studied variables, porosity (%P) was affected significantly (p <0.05) by the drying
cycles, composition, depressurization rate, and its interactions. The upper levels of these
variables showed a negative effect on the %P. The most porous materials (94.3-94.8%) were
obtained for AU samples dried using 1 cycle and depressurized at the lowest rate. The SBET
values for these AE were 127.6-192.6 m2 g-1. On the contrary, AUT samples dried using 2
cycles at the highest depressurization rate were less porous (79.9-81.6%) and had lower SBET
values (13.1-63.5 m2 g-1). Based on DP, both AE were mesoporous with values between 11.96-
17.37 nm, desirable for biomedical applications.
It is known that structure formation mainly takes place during the depressurization period. On
one hand, the drop of CO2 pressure (i.e., its expansion inside the AE and thus the stress in the
polymeric network) produced after 2 drying cycles in batch mode strongly affected the AE
structure in comparison with that produced after only 1 cycle. On the other hand, high
decompression rates lead to a rapid exit of CO2 from the AE producing many fissures and,
hence, low porosity.
The presence of urea and thiourea in the final AE was confirmed using FT-IR. These
compounds are capable of disrupting the native H-bonding of MCC destroying its crystalline
structure and establishing new bonds with -OH groups generating new networks. Both AE
showed a low degree of crystallinity, and high aqueous solution uptake (%A). AU presented
values of 400 %A while for AUT were considerably lower 250 %A, related to the higher
%P and SBET values obtained for AU. In this way, the AE structure obtained after scCO2 drying
process depended principally on the depressurization step and its chemical nature. AU showed
more desirable properties for biomedical applications such as bioactive loading and release.

Keywords: cellulose microcrystalline, aerogel, supercritical drying, porosity

References :
1 M. P. Dirauf, P. C. Wagner and A. S. Braeuer, J. Supercrit. Fluids, 2022, 191, 105762.
2 S. Zhang, F. X. Li, J. yong Yu and Y. Lo Hsieh, Carbohydr. Polym., 2010, 81, 668–674.

300
 Supercritical CO2-assisted impregnation of PLA films with
R-carvone. II: Effect of process variables on mechanical behavior

Florencia Muratore, Patricia P. Miranda, Lautaro Luján, Nicolás A. Gañan, María L. Goñi,
Raquel E. Martini*

IPQA (UNC – CONICET), Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina.

ICTA, FCEFyN, Universidad Nacional de Córdoba, Córdoba, X5016GCA, Argentina.
*corresponding author email address: raquel.martini@unc.edu.ar

GRAPHICAL ABSTRACT

ABSTRACT
Supercritical CO2-assisted impregnation has been investigated for several years as an emerging
technology for the incorporation of active compounds into polymers for different applications.
In particular, the incorporation of natural-occurring compounds with antimicrobial,
antioxidant, and/or insecticidal activities into flexible polymer films appears as an interesting
strategy to obtain active materials for food preservation. In addition, the food packaging
industry has shown an ever-increasing interest in the use of biodegradable materials, such as
polylactic acid (PLA), aiming to gradually decrease the use of petroleum-based polymers,
which nowadays represents an environmental problem. In this sense, the supercritical
impregnation of PLA films with R-(–)-carvone, a compound with known antimicrobial and
insecticidal activity, was studied as a route to develop an active material for food packaging
applications. The influence of process variables on the impregnation yield, release kinetics, and
thermal and crystallinity properties was previously studied [1,2]. In this work, a systematic
study of the effect of the supercritical processing and the incorporation of R-(–)-carvone on the
mechanical behavior of PLA films was assessed.
Commercial films of PLA were kindly provided by Converflex (ARCOR Group, Argentina),
while R-(–)-carvone (98% purity) was purchased from Sigma-Aldrich (Germany) and
industrial extra-dry carbon dioxide (water content ≤ 10 ppm v/v) was from Linde (Argentina).

301
The supercritical processing of PLA films with and without the addition of carvone was
performed in batch mode, using a lab-scale high-pressure system described before [1] Three
process variables were evaluated: scCO2 density (278-683 kg.m-3), temperature (40-60 °C),
and depressurization rate (0.6-6.0 MPa.min-1). All runs were performed in duplicate, using a
contact time of 2 h and a carvone/PLA mass ratio of 0 or 0.5.
Tensile properties were analyzed in a universal testing machine (Instron, USA), according to
ASTM D 882–12. The statistical effect of the process variables on elastic modulus (EM),
tensile strength (TS), and elongation at break (E%) was assessed by ANOVA testing (p < 0.05).
Neat PLA films showed a brittle behavior, presenting high EM (1606.7 ± 168.9 MPa), and low
E% and TS (E% = 11.6 ± 8.3 %, TS = 25.4 ± 2.0 MPa). The samples treated with scCO2, in
absence of carvone, presented similar tensile behavior (EM = 1285 – 1749 MPa), but with
higher TS (25.5 – 38.9 MPa) and E% (45.4 – 123.4 %) values. The increase in tensile strength
could be attributed to the higher crystallinity degrees of processed samples (~3% for neat
samples and ~25–30% for treated samples, as reported in the first part of this study [2]). On the
other hand, higher elongation values, opposite to that expected for higher crystallinities, can be
explained by the relaxation of tensions imposed by stretching during the PLA film
manufacturing, due to the plasticization induced by scCO2. ANOVA testing suggested that
depressurization rate has no statistical effect on the mechanical behavior of pressurized
samples. On the other hand, scCO2 density showed a significant effect on EM and TS,
decreasing their values for higher densities. Furthermore, temperature presented a significant
effect on TS and E%, decreasing both properties as temperature increases.
The incorporation of carvone was also studied, obtaining films with ~6–30 carvone wt% [1].
These samples showed higher E% and lower TS, compared to the pressurized samples without
the addition of carvone. The higher changes were observed at lower CO2 density, conditions
that favor the carvone impregnation. According to ANOVA testing, for these samples, scCO2
density showed a significant positive effect on all responses, increasing the value of EM, TS,
and E% for higher densities. On the other hand, the temperature presented a significant effect
only on EM and TS, decreasing both properties as temperature increases. Finally,
depressurization rate showed an effect only on EM and E% obtaining higher EM and lower E%
when fast depressurization is used. Furthermore, a decrease in EM was observed as the carvone
amount increased in the polymer matrix, probably due to an increase of free volume in the
amorphous phase introduced by the higher plasticizer effect due to the presence of carvone.
Particularly, EM suddenly decays at a carvone amount of 10 wt%, obtaining values around 200
MPa and lower than 50 MPa for higher additive content, suggesting that phase separation of
the additive starts at this concentration, as observed in the thermal behavior study [2]. Finally,
the results obtained here may provide useful information for broadening the knowledge of the
individual effect of the process variables and the amount of additive incorporated into the
polymer matrix on the mechanical behavior of PLA films.

Keywords: Supercritical solvent processing, mechanical properties, PLA, active packaging

References:
[1] P. Miranda et al., Journal of CO2 Utilization, 2022, 61:102029.
[2] P. Miranda et al., “Supercritical CO2-assisted impregnation of R-carvone into PLA films.
I: Effect of processing on thermal and crystallinity properties”, VI Prosciba, 2023.

302