Am 18 Ana Abstracts

ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018
ANA 1a: Spectroscopic, Spectrometric and Chemometric Methods for Lipid Analysis
Chairs: Sanjeewa Karunathilaka, US Food and Drug Administration, USA; and
Bernd W.K. Diehl, Spectral Service AG, Germany
Portable Raman Spectroscopy and Chemometric least-squares regression (PLSR) to predict fatty
Methods for the Analysis of Marine Oil Dietary acid composition. The current study evaluates
Supplements Betsy J. Yakes*, Sanjeewa R. the capabilities of each Raman spectrometer for
Karunathilaka, Kyungeun Lee, Lea Brückner, and the simple, rapid analysis of marine oil products
Magdi Mossoba, US Food and Drug for potential use in verifying label declarations
Administration, USA and quality control during dietary supplement
Marine oil supplements containing long-chain production.
omega-3 polyunsaturated fatty acids (PUFAs),
including eicosapentaenoic acid (EPA) and Vibrational Spectroscopy and Chemometric
docosahexaenoic acid (DHA), are the most Procedures for the Rapid Assessment of Olive
commonly used nonvitamin, nonmineral dietary Oil Authenticity Magdi Mossoba*, Sanjeewa R.
supplements in United States due to their Karunathilaka, Cynthia Srigley, Kyungeun Lee,
potential health benefits. While established Lea Brückner, and Betsy J. Yakes, US Food and
methods such GC-FID excel at analysis of omega- Drug Administration, USA
3 PUFAs, there is a need for rapid and accurate FDA is mandated with protection of the US
screening of these omega-3 PUFA products to public against intentional adulteration of foods
ensure both quality and accuracy of label for economically motivated gain, including having
declaration. As such, Raman spectroscopy using jurisdiction over deceptive label declarations
portable instruments followed by chemometric found with adulterated extra virgin olive oil
analysis was performed to understand the (EVOO). In April 2016, a US Congressional
potential for rapid, on-site evaluation of these Committee expressed concerns related to reports
supplements for fatty acid content. For this of the high prevalence of imported olive oil sold
study, 109 marine oils were procured, and in the US that is adulterated or mislabeled. Such
spectra acquired for each neat (underivatized) oil fraudulence reportedly includes the mixing of
on three Raman devices: (1) a portable Raman EVOO with seed oils, which could adversely
spectrometer with a unique laser setup and data impact the health of consumers who are allergic
processing to decrease fluorescence to seed oil. This issue underscores the urgent
interference, (2) a handheld Raman analyzer with need for more rigorous analytical methodologies,
onboard chemometrics for immediate pass/fail including untargeted rapid screening tools, for
triaging of complex samples, and (3) a smart- detecting such fraudulence. We have applied
phone sized Raman spectrometer containing rapid vibrational spectroscopy and chemometric
orbital raster scanning that allows for larger procedures to the screening of 72 retail
sampling areas. For each instrument, a broad- commercial products labeled extra virgin olive oil
based calibration library comprised of a wide for the classification, and/or prediction of volatile
variety of marine oils was made using partial content, fatty acid composition, and the type and
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quantity of a refined edible oil potentially mixed phosphatidylethanolamine (LPE) in lecithin and of
with authentic EVOO. The current spectroscopic/ PC, PC-ether, LPC, PE, APE, LPE in krill oil matrix.
chemometric predictions will be compared to The routine includes NMR spectra import,
those based on the International Olive Council extraction of data points, peaking of minima and
(IOC) official method COI/T.20/Doc. No 20/ maxima in the data, integration, quantitation
Rev. 3 (2010) for the determination of olive oil against internal standard, reporting of results as
purity; this method is based on the targeted Word file and their importing in our internal
determination of the absolute difference database. Our extensive studies on a
between the experimental HPLC values for TAGs representative set of more than 1000 lecithin
with equivalent carbon number 42 (ECN42_HPLC) (soy, raps, sunflower) and krill samples showed
and the theoretical value for TAGs with an that the automated routine can automatically
equivalent carbon number of 42 (ECN and accurately calculate the concentrations of all
42_theoretical) calculated from GC-based fatty PLs. No systematic or proportional differences
acid composition. between automated and manual evaluation were
detected. The developed program produces
Automated Multicomponent Phospholipid accurate results with the advantage of being fully
Analysis Using 31p NMR Spectroscopy: Example automated and requires less than 5 seconds for
of Vegetable Lecithin and Krill Oil Bernd W.K. each analysis. This tool is already used in high-
Diehl*, and Yulia B. Monakhova, Spectral Service throughput PL analysis of krill and lecithin and
AG, Germany
will be adjusted to other matrices (egg, milk,
Nuclear magnetic resonance spectroscopy
chocolate, etc.) as well.
(NMR) is widely applied in the field of
metabolomics due to its quantitative nature and Analysis and Detection of Olive Oil Adulteration
the reproducibility of data generated. However; using Fourier Transform Near-Infrared
one of the main challenges in routine analysis by Spectroscopy Ariel Bohman*1, Kathryn J. Lawson-
NMR is to obtain valuable information from large Wood2, and Robert Packer1, 1PerkinElmer, USA;
datasets of raw data in a high throughput, 2PerkinElmer, United Kingdom
automatic, and reproducible way. In this study, a Olive oil has seen a large increase in market
method to automatically annotate and quantify demand due to its linkage with lowering one’s
12 phospholipids (PLs) in vegetable lecithin (soy, risk of heart disease associated with its high
sunflower, raps) and krill oil is introduced. monounsaturated fat content. As the demand
Automated routines were written in MATLAB and value of olive oil increases, less reputable
environment for quantification of producers look for ways to cut costs and increase
phosphatidylcholine (PC), phosphatidylinositol profit margins resulting in the substitution or
(PI), lyso-phosphatidylcholine (LPC), dilution of olive oil with less expensive edible oils.
phosphatidylserine (PS), Economically motivated adulteration is the
phosphatidylethanolamine (PE), intentional addition of lower value substances to
diphosphatidylglycerol (DPG), a product to increase company profit margins.
phosphatidylglycerol (PG) and lyso- The prevalence of adulterated olive oil has been
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increasing with a reported 80% of Italian olive oil successfully distinguishing between olive oil and
being considered fraudulent. Near-infrared edible oils commonly used in the adulteration of
spectroscopy, for detection of olive oil olive oil. The objective of this work is to
adulteration, offers many advantages over demonstrate the utility of near-infrared
traditional reference methods, such as GC/MS spectroscopy coupled with chemometric
and HPLC, as it is a rapid, non-destructive methods in the detection of adulterated olive
technique. Samples of olive oil and commonly oils. Analysis of transmission spectra of pure and
used adulterants have been analyzed in adulterated olive oil samples will be presented to
transmission mode over the near-infrared region highlight the benefits of near-infrared
spanning from 14,000 to 4,000 cm-1. Near- spectroscopy as a rapid, non-destructive
infrared spectroscopy coupled with chemometric alternative to traditional screening methods for
analysis methods, such as soft independent the detection of olive oil adulteration.
modelling of class analogy (SIMCA), is capable of
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ANA 1b: Lipidomic Analysis

Chairs: Francesca Giuffrida, Nestec SA, Switzerland; and J. David Pinkston,
Kellogg Co., USA
Lipidomic Profiling–An Integral Technology for distinctive sensory profile and its reputation as a
Research and Development Elizaveta healthy source of dietary fats makes olive oil a
Freinkman*, Metabolon, Inc., USA target for adulteration by illegal blending with
Lipids are a diverse class of metabolites that other vegetable oils and deliberate mislabelling.
serve many biological functions such as energy The lack of efficient and harmonised analytical
storage, structural components of cell methods for detecting olive oil fraud has led to
membranes, and signaling. Accurate significant weaknesses that are exploited by
measurements of lipids are essential for counterfeiters. As a result, olive oil adulteration
biomarker discovery and for clarifying biological for the purpose of financial gain has become one
of the biggest sources of agricultural fraud in the
questions. This presentation will cover
EU. OLEUM is a collaborative 4 years EU project
Metabolon’s different offerings pertaining to lipid
(September 2016-2020) with 20 partners from 15
research, and these include the complex lipid countries. The project has 3 strategic objectives i)
panel, sebum lipid panel, stratum corneum lipid develop new and/or improved analytical
panel, and the global metabolomic profiling. The methods, ii) create the OLEUM Databank and iii)
applications of lipidomic profiling in research will establish the OLEUM Network. In this context,
be exemplified by case studies. Surveyor, the Fera has developed analytical methods to
complex lipid data visualization and analysis evaluate the quality and authenticity for a large
software tool will be briefly discussed as well. data set of olive oils. The quality will be
determined through volatile analysis with a focus
Non-targeted Analysis for Quality and on the relevant sensory defects (fusty, muddy-
Authenticity Determination of Olive Oil. James sediment, etc.) by a thermal desorption unit gas
A. Donarski1, Victoria Bailey-Horne1, Enrico Valli2, chromatography time of flight mass
Diego L. García González3, and Tullia G.T. Gallina spectrometer. The authenticity analysis will be
Toschi4, 1Fera Science Ltd., UK; 2University of based on phenolic compounds determined by
Bologna; 3Instituto de la Grasa (CSIC), Spain; liquid chromatography time of flight mass
4Alma Mater Studiorum—University of spectrometry (LC-TOF-MS) as a fingerprint
Bologna, Italy profile. This presentation will give an overview of
Europe is currently the largest producer of these developed non-targeted analytical
olive oil accounting for more than 70% of the methods, the related preliminary results and
world’s production. Non-EU countries are statistical data analysis.
expanding their domestic production and
increasing the competitiveness of the global olive
oil market. The high price of olive oil, the
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Supercritical Chromatography in Lipidomics practices. The global market for edible oils is
Applications: “Finally Ready for Prime Time?” expected to exceed 200 million metric tons by
Paolo Lecchi1, Yao Lu1, Erwin Kaal2, Rob Van der 2020, thus simple analysis of oil quality is
Hoeven2, and Dominik Burger1, 1DSM Nutritional essential to maintain process efficiency. Various
Products, USA; 2DSM Food Specialties, parameters are used to assess edible oil quality,
The Netherlands including iodine value (IV), free fatty acid (FFA)
Supercritical Fluid Chromatography (SFC) has
content, and numerous other parameters, as
all the premises for being an ideal component of
demonstrated in this submission. Traditional
analytical methods aim at characterizing complex
methods of analysis are typically carried out
lipid matrices. For instance, SFC has an operating
using standardized methods approved by the
range that suits the polarities of most of the lipid
American Oil Chemists’ Society (AOCS) and/or
structures and it allows faster separations with
the Germany Society for Fat Science (DGF).
lower operating costs. Moreover, SFC has been
However, these methods usually determine only
successfully interfaced to a variety of detectors,
one particular parameter and analysis is often
such as mass spectrometers equipped with ion
time-consuming. Additionally, sample
sources operating at atmospheric pressure.
preparation can be extensive, with hazardous
Despite these positive aspects, SFC is still not
solvents and reagents being required. Near-
frequently used in lipidomics applications. We
Infrared (NIR) spectroscopy is an already widely
will present insights and considerations on the
used technique in the food industry for
benefits and pitfalls encountered when
quantitative analysis of nutritional and quality
implementing analytical methods that include
parameters. Chemometric techniques, such as
SFC for the analysis of lipid extracts. This
partial least squares (PLS) used in conjunction
presentation will also include a direct comparison
with NIR spectroscopy, allow multivariate
between SFC-MS and other workflows for
analysis of samples by building a library of
lipidomics, and will emphasize the pros and
standards of pre-determined property values. In
cons of using SFC separation in MS-based
this submission, NIR spectroscopy with PLS
analytical platforms for the comprehensive
calibrations for the determination of quality
analysis of lipids.
parameters in various edible oils is
demonstrated. Oil samples are simply pipetted
A Rapid Non-destructive Method for
Determining Quality Parameters of Edible Oils into a fixed pathlength glass vial and measured in
Kathryn J. Lawson-Wood*1, Ariel Bohman2, and the transmission sample compartment of the
Robert Packer2, 1PerkinElmer, United Kingdom; spectrometer. Analysis is rapid, non-destructive,
2PerkinElmer, USA simple to undertake and allows multiple variables
Edible oils are an integral part of human to be determined instantaneously.
diets; being using in virtually all types of culinary
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ANA 2a: Analysis of Fats and Oils Applying Advanced Lipid Analysis Techniques
Chairs: William C. Byrdwell, USDA, ARS, BHNRC, FCMDL, USA; and Walter Vetter,
University of Hohenheim, Germany
Use of Countercurrent Chromatography (CCC) Conclusion. CCC can be successfully used for the
for the Preparative Isolation of Lipid Compounds isolation of valuable lipid compounds in the
Walter Vetter*, Marco Müller, Katharina medium mg-range (per run). Isolated lipid
Wasmer, Andrea Goncalves Peca, and Medisa compounds can be used to investigate their
Muric, University of Hohenheim, Germany biological activity or as reference standards for
Objective. Countercurrent chromatography quantitative analyses.
(CCC) is a preparative, all-liquid based
chromatographic method predominantly used for Investigation of Olive Oil Substitution with
the isolation of natural compounds. Separation is Other Edible Oils by Ultra High-Performance
obtained by means of two immiscible liquids, i.e. Liquid Chromatography Separation of
the solvent system, inside a hollow tube. The Triglycerides Pierluigi Delmonte* and Andrea
solvent system is permanently mixed and de- Milani, US Food and Drug Administration, USA
mixed inside the so-called CCC centrifuge which In recent years several claims have been
substitutes the column of an otherwise similar made regarding the substitution in the
periphery of CCC and HPLC. Development of marketplace of olive oil with edible oils of lower
suitable solvent systems is the key-factor for commercial value. Olive oil, produced from the
successful CCC applications. Limitations in the pulp of the fruit, is characterized by a distinctive
use of CCC exist especially in the nonpolar range triglycerides (TAG) composition compared to
of lipid compounds. Methods Used. During the seed oils. The difference between the theoretical
last years we have developed solvent systems and experimentally determined content in TAG
(e.g. the BTF system) and used new or rare CCC with equivalent carbon number (ECN) 42 is the
modes (heart-cut-2D-CCC, recycling mode, co- most widely applied parameter to detect this
current mode, multiple injection mode) for the economic adulteration. The incomplete TAG
isolation of different lipid compounds. Results. separation provided by traditional high-
Typically, ~1 g lipid sample was injected and ca. performance liquid chromatography (HPLC)
5–100 mg of the target compound were isolated conditions applied to the determination of the
per run (run time ~1–3 h), in dependence of its delta-ECN 42 value limits the ability of detecting
contribution to the sample and the required sophisticated frauds. In this work the TAG
purity. Recent successful applications (isolation separation is achieved by utilizing multiple ultra-
of tocotrienols, tocomonoenols, amyrins, furan high-performance liquid chromatography (UPLC)
fatty acids, etc.) will be shown with focus on the separation columns in series, obtaining the
proper selection of starting materials, solvent separation of almost all major TAGs contained in
systems and special CCC modes. As a further olive oil. Identification of individual TAGs
benefit, CCC fractionation allows to detect minor separated by UPLC is achieved by LC-GC in offline
lipid compounds which are usually overlooked. mode. The detailed TAG compositions of
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available reference olive oils and potential seed tocopherol as the internal standard. In the
oils adulterants allow the determination of more second dimension, TAG isomers are separated
sophisticated parameters beyond the delta-ECN according to double bond-containing fatty acids
42 for detecting the partial or complete in different positions on the glycerol backbone
substitution of olive oil with other edible oils. (i.e., regioisomers).
Comprehensive Dual Liquid Chromatography Development of Lipidomics-based Reference

with Quadruple Mass Spectrometry, LC2MS4, Materials and Reference Data for Oils
for Jacaranda Mimosifolia Triacylglycerols John Bowden*, National Institute of Standards
William C. Byrdwell*, USDA, ARS, BHNRC, and Technology, USA
FCMDL, USA With a growing number of new lipidomic
Online comprehensive two-dimensional users, instrumentation, and applications, efforts
liquid chromatography has been combined aimed to dissect the finer details of lipid
with quadruple parallel mass spectrometry for measurement and improve community-wide
LC1MS2 x LC1MS2 = LC2MS4 experiments. harmonization are imperative. This presentation
Two mass spectrometers are used for qualitative will highlight recent efforts by the National
and quantitative analysis during non-aqueous Institute of Standards and Technology (NIST) to
reversed-phase liquid chromatography (NARP-LC) improve lipidomic measurement by promoting
as the first dimension, in addition to a UV the creation and implementation of oil-based
detector, fluorescence detector (FLD), reference materials and reference data. Initial
evaporative light-scattering detector (ELSD), and efforts have focused on human health and
corona charged aerosol detector (CAD). High- consumer-based reference materials. Using
resolution accurate-mass (HRAM) electrospray Standard Reference Material (SRM) 2378 (serum
ionization mass spectrometry (ESI-MS) on an from donors who took fish or flaxseed oil
Orbitrap instrument was used in parallel with supplements), an interlaboratory comparison
atmospheric pressure chemical ionization (APCI) exercise for lipidomics was executed, resulting in
MS on a tandem sector quadrupole (TSQ) determining the extent of agreement present in
instrument in the first dimension. Two additional current lipidomic measurement within the
mass spectrometers plus UV detection are used community, the formation of consensus means
for qualitative analysis of the second dimension, with associated uncertainties for lipids present,
which employs a lab-made silver-ion column for and the identification of challenges associated
UHPLC. Atmospheric pressure photoionization with current lipid measurement. The
(APPI) TSQ MS and ESI-MS were used in parallel presentation will also highlight new efforts to
for the second dimension. In the first dimension, provide reference materials and reference data
diacylglycerols (DAGs) and triacylglycerols (TAGs) for the analysis of olive, argan, and krill oil. The
in Jacaranda mimosifolia (blue jacaranda) are utility of high resolution mass spectrometry-
reported as relative percentage composition, based lipidomics will be addressed for the
while alpha-tocopherols and gamma-tocopherol characterization of oils, in concert with the
are quantified by calibration lines, using d6-alpha-
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formation and community-wide application of The Hybrid Search: A New Mass Spectral Library
oil-based “sample-method-data” products. Search Approach for Compound Classification.
Arun S. Moorthy1, Brian T. Cooper2, William E.
A Rapid and Efficient Method for Lipid Wallace1, and Stephen E. Stein1, 1National
Separation with Supercritical Fluid Institute of Standards and Technology, USA;
2University of North Carolina at Charlotte, USA
Chromatography Coupled with High Resolution
Mass Spectrometry. Sheher Moshin, Agilent Compound identification is a key task across
Technologies, USA numerous chemical application areas, including
A method was developed for rapid separation but not limited to food, pharmaceutical,
of a broad range of lipids based on Supercritical petroleum and lipidomics. Mass spectral library
Fluid Chromatography (SFC) and Electrospray searching has become one of the primary
Ionization (ESI) Quadrupole Time of Flight Mass analytical techniques employed in compound
Spectrometry. The method allows separation of identification. The limitation of most library
a broad range of lipid classes in less than 15
search algorithms is the requirement that the
minutes. This approach offers distinct
mass spectral database contain a representative
advantages of comparative UHPLC methods in
terms of speed and resolution. spectrum of the query compound (analyte). This
Results will be shown for optimization of limitation is particularly restrictive when novel
separation, acquisition and detection molecules are synthesized (or discovered in the
parameters. Several columns including the new case of lipidomics) more rapidly than they can be
Hilic Zwetterion column, mobile phases, mobile added to mass spectral libraries. In this
phase additives, flow rates and column presentation, we describe the new “Hybrid
temperatures have been evaluated for best Search” method that combines fragment-ion and
inter/intra class lipid separation. Data acquisition neutral-loss matching when computing similarity
parameters with data dependent acquisition of match factors. In doing so, the search algorithm
lipid samples have been evaluated with the goal generates high scores for compounds that differ
of maximizing the number of detectable lipids. by a single chemical modification yet share
Results will be presented for biological
similar fragmentation behavior (chemical
samples. The entire workflow including data
cognates). Accordingly, if an analyte is NOT
analysis with feature extraction, spectral
identification with Simlipid and statistical analysis contained within the library, it can still be
with Agilent Mass Profiler Professional will be correctly classified given that its chemical
shown. cognates are contained in the library. Following a
detailed description of the algorithm, numerous
example applications are presented, including
the identification of glycerophospholipids.
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ANA 2b: Olive Oil, including Sensory Analysis

Chairs: Selina C. Wang, University of California-Davis, Olive Center, USA; and Susan Seegers,
Bunge North America, USA
Contribution of Flavor Compounds to Explain “strawberry-like” perceptions, and the other type
New Sensory Defects in Virgin Olive Oil: The characterized with “wood” and “humidity”
Example of “Frostbitten Olives” Diego L. García descriptors. Different volatile compounds
González1, Inmaculada Romero1, Ramón explained these differences.
Aparicio-Ruiz1, Noelia Tena1, Ana Lobo1,
María Teresa Morales2, and Aparicio Ramón1, The Profitable Relation between Sensory and
1Instituto de la Grasa (CSIC), Spain; 2University
Analytics in Virgin Olive Oil Quality Detection.
of Seville, Spain Tullia Gallina Toschi1, Sara Barbieri1, Chiara
The aroma of virgin olive oil determines, Cevoli1, Ole Winkelmann2, Karolina Brkić Bubola3,
together with other parameters, the quality Florence Lacoste4, Milena Bučar-Miklavčič5,
designation and therefore its price in the market. Ummuhan Tibet6, Ramón Aparicio-Ruiz7,
The strict control of virgin olive oil by means of Diego L. García González7, and Alessandra
the organoleptic evaluation has contributed to Bendini1, 1DISTAL University of Bologna, Italy;
2Eurofins Analytik GmbH, Germany; 3Institute
the fact that the quality of this product on the
market has reached high standards. The current of Agriculture and Tourism, Croatia; 4Institut
des Corps Gras, France; 5Science and Research
knowledge on volatile compounds and sensory
Centre Koper, Slovenia; 6Ulusal Zeytin ve
analysis leads to new opportunities to improve
Zeytinyağı Konseyi, Turkey; 7Instituto de la
the performance of panel tests and to
Grasa (CSIC), Spain
understand why some sensory defects are To meet the expectations of the consumers
sometimes present and the technological reasons and preserve the image of virgin olive oils
for them. An extensive research has been carried (VOOs), it is necessary to guarantee its quality
out on the characterization of those volatiles and authenticity. Despite many methods to
responsible for the main sensory defects (rancid, detect VOO fraud and assess its quality are
fusty, mustiness-humidity, winey-vinegary). available at international level, the need of
Other sensory defects, which were less usual only supporting the sensory analysis was reported in
a few decades ago, are becoming much more the call H2020-SFS-14a-2014 and is one of the
common today due to a change in the weather main objective of the OLEUM project.
pattern in the last few years. One of them is the Specifically, the setting up of rapid instrumental
off-flavor called “frostbitten olives” produced as methods could represent a solution for achieving
a consequence of freeze injuries in olives. In this a preliminary screening of a high number of
study we address the analysis of the volatile samples and/or for supporting the sensory panels
composition of virgin olive oils characterized with in the discrimination of boundaries samples. In
this defect. The volatile information allowed this research, a set of 180 VOOs, sampled during
characterizing grouping these oils into two types, the first year of the OLEUM project, was sensory
one of them characterized with “soapy” and assessed (EU Reg. 1227/2016). The sensory
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evaluation was performed by 6 sensory panels, determinations. Thus, in this study, LC-MS and
having different nationality and experience. The GC-MS platforms were used to develop two new
headspace volatile profiles of this set, analyzed methodologies capable of carrying out the
by many chromatographic techniques, were also simultaneous determination of more than 40
analyzed by flash gas chromatography electronic compounds belonging to different VOO minor
nose (FGC E-nose). The chromatograms were chemical classes within a single run. A non-
elaborated by multivariate statistical analysis to selective and highly efficient liquid-liquid
develop a classification model able to quickly extraction protocol was optimized for VOO minor
discriminate between “extra virgin” and “no components isolation. The separation and
extra virgin” olive oils. This approach could detection conditions were adjusted to determine
reduce the number of samples to be assessed by phenolic and triterpenic compounds, free fatty
a sensory panel and decrease the number of acids and tocopherols by LC-MS, plus sterols and
cases giving rise to an uncertain or debated hydrocarbons by GC-MS. A comparative
classification. This work was developed in the assessment of both methods in terms of
context of the project OLEUM “Advanced analytical performance, easiness, cost, and
solutions for assuring authenticity and quality of adequacy to the analysis of each class was
olive oil at global scale” funded by the European performed. The new methods were then applied
Commission within the Horizon 2020 Programme to study 126 oil samples from six different
(2014–2020, G.A no. 635690). Mediterranean PDOs (Meknès and Ouazzane
(Morocco), Priego de Córdoba and Baena (Spain),
Deep Insight Into the Minor Fraction of Virgin Kalamata (Greece) and Toscana (Italy)). Data
Olive Oil by Using New LC-MS and GC-MS treatment implied the selection of molecular
Multi-class Methodologies: Application to features, bucketing, filtering, scaling and
Discriminate Samples from Different Protected normalization; these steps as well as the
Designations of Origin Alegria Carrasco-
application of principal components analysis and
Pancorbo1, Lucía Olmo-García1, Juan J. Polari2,
partial least square-discriminant analysis to the
Xueqi Li3, Aadil Bajoub1, Karin Wendt4, Nikolas
Kessler4, Carsten Baessmann4, Alberto LC-MS and GC-MS data were done by using
Fernández-Gutiérrez1, Selina C. Wang2, 3, and MetaboScape® software. Noticeable
Alegría Carrasco-Pancorbo1, 1Dept. of Analytical discrimination among the six evaluated PDOs was
Chemistry, Faculty of Sciences, University of achieved taking into account the data coming
Granada, Spain; 2Dept. of Food Science and from both platforms. The contribution of a few
Technology, University of California Davis, USA; thousand molecular features to the statistical
3Olive Center, University of California Davis, USA;
models was evaluated in depth and several
4Bruker Daltonik GmbH, Germany
compounds such as elenolic acid,
The emergence of multi-class analytical acetoxypinoresinol, oleuropein and ligstroside
methodologies might greatly increase throughput aglycones, and some other tentatively identified
and reduce cost, while avoiding the complexity substances were pointed out as possible PDOs
and redundancy of single-chemical class distinctive markers.
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1H NMR–metabolic Profiles of Monocultivar Coco L., Mondelli D., Mezzapesa G.N., Miano T.,
EVOOs for PDO, PGI and 100% Italian Blend De Pascali S.A., Girelli C.R., Fanizzi F.P. (2016) J
Production Assessment Chiara Roberta Girelli, Am Oil Chem Soc 93: 373-381. Girelli, C.R., Del
Laura Del Coco1, Federica Angilè2, and Francesco Coco, L., Fanizzi F.P. (2016) Eur J Lipid Sci Tech
Paolo Fanizzi*2, 1Department of Biological and 118: 1380–1388. Girelli C.R., Del Coco L.,
Environmental Sciences and Technologies Fanizzi F.P. (2017) Sustainability 9: 1471
(Di.S.Te.B.A.), University of Salento, , Italy;
2Dipartimento di Scienze e Tecnologie Biologiche and references therein
ed Ambientali Università del Salento, Italy
“Musty”, “Fusty, Muddy Sediment”, and
Mediterranean food tradition is based on
“Rancid” Off-flavors in olive Oils are Well-
three basic essentials: wheat, olives and grapes.
known: but what is Behind on a Molecular
Nevertheless, olive oil is the central element for Level? Michael Granvogl*, Anja Neugebauer,
its increasing consumption around the world. The and Peter Schieberle, Technical University of
need of a scientific tool to assess EVOOs Munich, Germany
geographical origin represents a hot topic issue, Extra virgin olive oils (EVOO) are very popular
since the EU Regulation 182 of 6 March 2009 (on edible oils because of the valuable nutrients and
the compulsory labeling of EVOOs with the flavors. The aroma of a food is one of the most
geographical origin of the olives) still lacks an important criteria for consumers’ buying
official validation methodology. The same is for behavior. Thus, a detailed knowledge of the
the EU Regulation 1151/2012, which seeks to respective key odorants is of high interest and,
enhance Europe’s agricultural quality policy by even more important, of the molecules causing
increasing the coherence of various high-quality distinct off-flavors, known as “musty”, “fusty,
products, such as EVOOs, marketed under a PDO muddy sediment”, and “rancid”. The molecular
and PGI labels. We are currently involved in sensory science concept was used to clarify the
several NMR-based metabolomics and formation of these off-flavors in EVOOs using a
chemometric studies to assess cultivar positive control showing the desired sensory
composition and geographical origin of extra attributes and oils eliciting the abovementioned
virgin olive oils, in particular for PDO, PGI and sensory defects. This methodology is based on
100% Italian blend EVOOs production (mainly comparative aroma extract dilution analysis via
from Apulia and Tuscany regions) [1]. A large- gas chromatography-olfactometry, followed by
scale analysis of 100% Italian blend and identification experiments via gas
monocultivar oils from specific geographical chromatography-mass spectrometry. The most
areas was performed by 1H NMR spectroscopy potent aroma compounds are quantitated by
together with multivariate statistical analyses. stable isotope dilution assays and odor activity
Our results confirmed the usefulness of suitable values (ratio of concentration to respective odor
monocultivar EVOO samples, possibly genetically threshold) are calculated. Finally, aroma
certified, for the construction of 1H NMR simulation experiments are performed to
metabolic profiles databases aimed to cultivar validate the analytical data. In addition, the
and/or geographical origin assessment. [1] Del identified key odorants were quantitated in
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further EVOOs and in olive oils with one of the pathway, will be discussed in the lecture. In
analyzed off-flavors to put the obtained results summary, the “musty”, “fusty, muddy sediment”,
on a broader basis, e.g., by principal component and the “rancid” off-flavors in EVOOs were
analysis. Main differences were obtained for characterized for the first time on a molecular
esters, acids, and phenolic compounds. Their level using the combination of an analytical-
formation, caused for example by instrumental, sensorial, and statistical approach.
microorganisms via the so-called Ehrlich
– 12 –
ANA 2c / LOQ 2a: Evaluation and Prediction of Oxidative Stability and Shelf-life
Chairs: Min Hu DuPont Nutrition & Health USA; and Rick Della Porta Pepsico / Frito-Lay USA
The Combination of High Oleic Oils and Natural (TPC), 2', 2-diphenyl-1-picrylhydrazyl (DPPH)
Antioxidants as a Powerful Tool for Shelf Life radical scavenging ability and oxygen radical
Extension Susan Knowlton*, DuPont Company, absorbance capacity (ORAC), and striped soybean
Pioneer, USA oil-in-water emulsion system were employed to
High oleic oils from soybean, canola, and evaluate the antioxidative activity of each
sunflower have made significant nutritional and fraction. LC-QTOF-MS was employed for
stability improvements in food manufacturing as identifying the structure change of soluble bound
they replace both partially hydrogenated and moieties which can explain the variation of the
conventional (high polyunsaturated), commodity antioxidative activity. Molecular weight of SBPCs
oils. Despite their natural stability resulting from tended to increase during chickpea germination.
a low polyunsaturated fatty acid content, some Higher molecular weight soluble phenolic
manufacturers require further improvements in compounds had greater antioxidative activity in
shelf life stability as they replace synthetic emulsion system. Some of the structures of
antioxidants in their formulations. Traditional SBPCs were identified by LC-QTOF-MS, which
antioxidants like TBHQ, BHA, BHT, and others may be related to the difference of antioxidative
have fallen into disfavor as consumers are activity in each fraction. The findings can be
demanding shorter, simpler, and more ‘natural’ applied to exploit antioxidants with strong
ingredients on food packaging labels. The antioxidative efficacy in food systems.
combination of high oleic oils with natural
antioxidants is a valuable tool to meet these Antioxidant Activities of Sugars and Protein in
demands. Data showing the extension of shelf Low Moisture Cracker System Thanh P. Vu*, Lili
life achieved with these combinations will be He, D. Julian McClements, and Eric A. Decker,
presented. University of Massachusetts Amherst, USA
In American diet, low moisture foods, such as
crackers, are a top saturated fat contributor
Chickpea Germination Improves the
Antioxidative Activity of its Soluble Phenolic exposing a potential risk of coronary heart
Compounds. Minwei Xu*, and Bingcan Chen, disease for American consumers. Replacing
North Dakota State University, USA saturated fat by unsaturated fat could improve
Soluble bound phenolic compounds (SBPCs) the nutritional properties of crackers, yet this
extracted from germinated chickpea (Cicer would require antioxidant strategies to prevent
aretinium L.) were fractionated using SEC-HPLC lipid oxidation and maintain the shelf life of the
coupled with auto fraction collector regarding to product. In this study, reducing sugars were
the molecular weight variation. Three fractions found to increase oxidative stability in crackers.
categorized by molecular weight were collected Glucose, maltose and maltodextrin at the same
at the optimum germination time (6 days). In dextrose equivalence resulted in increased
vitro assay, including total phenolic content hydroperoxide (30, 48, and 48 days, respectively)
– 13 –
and hexanal lag phases (60, 81, and 78 days, like a 40% fat spreads contains transition metal
respectively), compared to control cracker (9 day ions? An antioxidant working well in a O/W food
hydroperoxide lag phase and 33 day hexanal lag emulsion would work efficiently in an W/O
phase). The antioxidant activity of maltose was emulsion? The presentation will highlight
effected by water activity (aw) and different evaluation methods being used to
concentrations. At aw values of 0.22 and 0.68, assess the oxidative stability of butter fat,
maltose increased hydroperoxide and hexanal lag spreads and margarines, investigate the impacts
phases, compared to its activity at aw 0.05. For of different antioxidant blends on the oxidative
example, crackers with 5.4% maltose at aw = 0.22 stability of spreads with varying fat contents, and
and 0.68 had 63 day and 186 day hexanal lag study how chelators could impact the oxidative
phases, respectively, compared to 48 days at aw stability of different spreads containing varying
= 0.05. Gluten addition from 2.5 to 10%, at aw = levels of fat.
0.05, did not affect oxidative stability, but at aw =
0.68 it increased hexanal lag phases. Casein only Shelf-life Extension of Meat and Meat Products
slightly increased hexanal lag phase at 7.5 and by Using Natural Antioxidants Henna FS Lu*,
10%, aw = 0.22. The ability of reducing sugars Kalsec Europe Ltd, UK
with low sweetness (e.g. maltose) to inhibit lipid Recently, the use of natural antioxidants has
oxidation in model crackers suggests that they gained attention globally as the result of rising
could be an effective antioxidant strategy. consumer demand for clean label solutions. Due
to the presence of unsaturated fat in membrane
Oxidative Stability of Margarines, Shortenings phospholipids, lipid oxidation occurs in meat
and Spreads Min Hu*, DuPont Nutrition & during processing and storage. Oxidative stability
Health, USA of various meat and meat products including
Oxidative stability and shelf life of food ground beef, British sausage and chicken fillets
emulsions like mayonnaise and salad dressing were conducted. Meat and meat products were
have been widely studied. However, the prepared with and without inclusion of
oxidative stability of W/O food emulsions such as antioxidants, followed by storage at different
margarines, shortenings and spreads have not temperature and packaging condition. The
been thoroughly studied yet, and the researches stability was monitored through measurements
on this area have started drawing a great deal of such as visual observation, color measurement,
attention in both industry and academia. Usually, sensory evaluation and secondary volatile
a bulk oil or fat may be more oxidatively stable oxidation profile by GC-MS (SPME). The effect of
than an O/W emulsion containing the bulk oil or synthetic antioxidants such as BHA, BHT, Sodium
fat. How about the case when comparing an O/W Metabisulphite and ascorbic acids versus natural
food emulsion with a W/O food emulsion? antioxidants such as rosemary, green tea, vinegar
Prooxidant transition metal ions like ferrous iron and acerola on the oxidative stability of meat and
have a big impact on oxidative stability of O/W meat products were investigated. Results showed
food emulsions such as mayonnaise and salad that inclusion of natural antioxidants significantly
dressing. What is the case when a W/O emulsion improved the shelf life of meat products. A better
– 14 –
color protection, better sensory attributes and a Antioxidant Testing – An Application Review.
lower level of oxidation compounds were Rick Della Porta*, Pepsico/Frito-Lay, USA
observed in products with antioxidants added. In
addition, different secondary volatile oxidation A review of the practices and applications of
profiles were obtained for different meat testing antioxidants on edible oils. The challenge
between academic research and practical
products. In short, natural antioxidant showed a
application is critical for getting a true
similar performance as compared to that of
understanding of how and antioxidant may be
synthetic antioxidant and therefore provide an used. The test protocols and analysis methods
alternative to replace synthetic antioxidant. The are reviewed in general terms in hope of making
finding of these studies provide valuable a better connection between researchers and
information and new insights to meat suppliers end-users.
to improve the shelf life of their meat and meat
products.
– 15 –
ANA 2d/LOQ 2b: Sensory Analytics and Analytical Methods

for Assessing Lipid Oxidation and Shelf-life
Chairs: Jian Kong, Abbott Nutrition, USA; and Richard Della Porta, Frito-Lay, USA
Antioxidant Efficacy and Impact of Storage condition. Based on this study, we confirmed that
Conditions Marie Shen1, Lan Ban1, and Chandra accelerated conditions can serve as quick
Ankolekar*2, 1Kemin Food Technologies, USA; reference tools but it is needed to establish
2Kemin Industries Inc., USA
ambient models that both the activity and
Oxidative stability of oils can be measured by stability of an active compound would serve as
accelerated studies such as OSI, Schaal oven, or independent parameters.
by monitoring real time chemical and sensory
changes at ambient condition. Most of the Sensory Directed Chemical Analysis of Oxidized
literature have depended on accelerated Marine Oils Roy D. Desrochers*, Tufts University
conditions for evaluating antioxidant efficacy for Sensory and Science Center, USA
the relative ease and quick results. However, Abstract not available.
debates over the reliability and accuracy still
exist. And it is not clear whether complex mixture Developing a Sensory Oxidation Quality Scale
as plant extracts behave the same way as Monica L. Godbout*, Abbott Nutrition, USA
synthetic antioxidants. We hope to answer these Abstract not available.
questions with full analyses of free radicals,
peroxides, secondary oxidative byproducts and Assessing Virgin Olive Oil Stability and Shelf Life
active compound degradation in the treated oils at Moderate Conditions by FTIR Spectroscopy
that are stored in both accelerated and ambient Endowed with a Mesh Cell Accessory Noelia
conditions. In this study, common ingredients Tena1, Ramón Aparicio-Ruiz1, Ana Lobo2, María
including TBHQ, rosemary extract (RE), mixed Teresa Morales3, Aparicio Ramón2, and Diego L.
tocopherols (MT) and oil soluble green tea García González*1, 1Instituto de la Grasa (CSIC),
extract (OSGT) were tested. Comparing one-year Spain; 2Instituto de la Grasa (CSIC); 3University of
ambient storage and 30-day accelerated storage, Seville, Spain
it is showed that although the absolute values for Virgin olive oil (VOO) stability is one of the
each test parameter were not the same, the major topics today because producers are
trends in antioxidant efficacy were every similar. demanding more effective methods to guarantee
For example, OSGT showed strong dose response that oils from extra virgin category remains in
in both conditions and performed better than RE this high-quality category during all the shelf life.
and MT. However, the relative degrees of However, moderate conditions of light and
improvement varied for RE and OSGT, that under temperature are sometimes enough to produce a
ambient storage, their relative performances quality loss in a few months. The current
were better. This improvement positively methods (e.g., Rancimat) apply high
correlated with the improved stability of the temperatures and, therefore, their results are
active compounds under ambient storage hardly correlated with the real conditions of
– 16 –
storage. In this context, mesh cell has been stability determined with Rancimat. The FTIR
proposed as a rapid tool designed to monitor spectra revealed a remarkable increase of
chemical changes that occur as a consequence of hydroperoxides and the subsequent formation of
oxidation at moderate conditions by Fourier secondary oxidation products (e.g., alcohols). The
transform infrared (FTIR) spectroscopy. In order fact that this method measures the stability of
to evaluate this approach, monocultivar VOOs the oil from a multi-factor perspective
have been stored in mesh cells under different (temperature and light) and includes several
temperatures (at 23, 35, 65 °C) and different light chemical species (primary and secondary
intensities (400, 1000, 7000 lx) simulating the oxidation products) makes the results more
real conditions during storage and transport. The reliable to optimize VOO handling (e.g.,
oil stability of the samples determined by using packaging and storage temperature) according
this accessory has been compared with the oil to the real conditions of storage.
– 17 –
ANA 3: General Analytical

Chairs: Torben Küchler, Eurofins Analytik GmbH, Germany; and Pierluigi Delmonte,
US Food and Drug Administration, USA
Rapid Identification and Relative Quantification classes by polarity with column chromatography
of the Phospholipid Composition in Commercial or liquid-liquid extraction. To avoid this time
Lecithins by 31P-NMR Ying Yang*, Richard consuming manual operation step, an alternative
Hiserodt, and Jing Li, International Flavors & is the selective separation of target analytes via
Fragrances Inc., USA HPLC. Combined with an on-line coupled GC
31P NMR analysis of samples prepared in a
system, rapid and robust methods can be
sodium cholate detergent system was used as a achieved. The presentation shows an overview
method for the identification and quantification about different applications for the evaluation of
of enzymatic hydrolysis products of lecithin. quality and authenticity of fats and oils with the
To precisely characterize all of the hydrolysis LC-GC technique: Distribution and content of
products from commercial lecithin, a series of sterols for the judgement of identity and
enzymatic reactions of each phospholipid with authenticity, detection of steradienes for the
phospholipase PLA1 were conducted and proof of a heat treatment of virgin oils, content
monitored by 31P NMR at different times. Twenty of fatty acid ethyl esters in olive oil as a marker of
six phosphorus-containing hydrolysis products poor raw material quality and determination of
from six classes of phospholipids (PC, PI, PS, PE, PAH as an important contaminant with a special
PG, PA) were found and determined by 31P NMR LC-LC-GC application. An outlook is given for
measurement. The impact of pH on the chemical possible future applications.
shift values for these hydrolysis products was
observed and reported. To our best knowledge, Overcoming Issues and Challenges in the
this is the first report of 31P NMR chemical shift Analyses of Tocols in Oils Mei Han Ng*
values for entire lyso-phospholipids hydrolyzed and Ahmad Kushairi Din, Malaysian Palm
from 6 classes of phospholipids. Rapid and Oil Board, Malaysia
routine analysis of phospholipid composition in Accuracy and reliability of analyses for tocols
commercial lecithins by 31P NMR was achieved (tocopherols, tocomonoenol and tocotrienols)
without the need of phospholipid standards. have always been a cause for concern as the
tocols are highly potent antioxidative compounds
Applications for the LC-GC Technique in Routine and thus, are easily susceptible to oxidation or
Fat and Oil Analysis Torben Küchler*, Eurofins degradation. The scarcity of tocotrienols and
Analytik GmbH, Germany tocomononeol standards also contributed to the
A standardized and robust sample difficulty in analyses and calibrations. Official
preparation is the basic requirement for a AOCS method recommended the use of α-
reliable analytical method. Many sample tocopherol as reference in the absence of
preparation methods in the fat and oil analysis tocotrienols standards, which is a common
depend on the selective separation of lipid practice. However, this is often not accepted by
– 18 –
industry players due to the question on the Methods Used: Extracted lipids were
different response of the individual tocopherols fractionated by SPE into (i) hydrocarbons, (ii)
and tocotrienols in UV spectroscopy, resulting in tocopheryl esters and (iii) free tocopherols. The
discrepancy in analyses. In addition, the separated tocopheryl ester fraction was analyzed
concentration of tocomonoenol is often not by GC/MS in intact form and after saponification
reported although its presence is significant in and silylation. Reference standards of alpha- and
gamma-tocopheryl esters were synthesized for
palm oil. A chromatographic method that is able
quantitation. In vitro incubation experiments
to address all the concerns faced in the analyses
were performed with artificial digestion juices.
of tocols need to be developed. This paper
reports on the development of a comprehensive Results: Tocopheryl esters were detected and
and reliable method for the HPLC analyses of quantified in four matrices (bell pepper, chili
pepper, cucumber, walnuts). Both the tocopherol
tocopherols, tocomonoenol and tocotrienols in
pattern and the absolute amounts were varied.
oil. Limit of detection, limit of quantitation, linear
In all four matrices, the concentration of the
range of analyses as well as the calibration of all tocopheryl esters was higher than the
the individual tocols, including tocomonoenol, in corresponding concentration of the free
the absence of authentic standards are taken into tocopherols. Subsequent incubation with
consideration. Crosscheck of the method artificial digestion juices showed that tocopheryl
developed with various laboratories showed that esters were not cleaved into free tocopherols
the method is accurate with good reproducibility. and fatty acids. Hence, tocopheryl esters do not
This method may be adopted as the official directly contribute to the tocopherol activity of
method for the analyses of tocols in the future. the food samples.
Conclusions: The bioactivity and possible
Tocopheryl Esters—Analysis of Novel Vitamin E contribution of tocopheryl esters to the vitamin E
Conjugates in Vegetable Foods: Occurrence, activity needs to be assessed given the
Concentrations and Digestibility Walter Vetter*, comparably high abundance in the tested food
Stephanie Krauß, and Vanessa Darwisch, matrices. More data is required on the role of
University of Hohenheim, Germany tocopheryl esters in foods.
Objective: Vitamin E comprises a group of mainly
four tocopherols (alpha-, beta-, gamma- and A Method for Detection of Partially
delta-T) along with four tocotrienols and a few Hydrogenated Oils (PHO) in Food Matrices
minor compounds. Tocopherols are typically Containing Vegetable Oils Sneh Bhandari1,
considered to occur in their free form in foods. Ming Gao1, and Pierluigi Delmonte2,
1Merieux Nutrisciences, USA; 2US Food
Recently, we noted the presence of tocopheryl
esters with different fatty acids in red and yellow and Drug Administration, USA
bell pepper. In this study, we screened further Foods on sale in the U.S. are being
vegetable foods and present a method for the reformulated to minimize the content of partially
quantification of tocopheryl esters in foods. hydrogenated oils (PHO) among their ingredients.
Recent determinations and regulations regarding
PHOs in foods indicate the necessity of
distinguishing between the trans fatty acids (TFA)
– 19 –
produced during oil refining and partial Supplementation Studies Involving Natural trans
hydrogenation. While most TFAs occurring in Fatty Acids: Real Technical Challenges, Actual
PHOs are also present in refined oils, their Solutions Etienne Guillocheau*, Daniel Catheline,
distribution pattern and concentrations are Philippe Legrand, and Vincent Rioux,
different. In this study we propose a Agrocampus-Ouest, France
methodology for detecting the occurrence, in Objectives and hypothesis: Growing evidence
foods, of TFAs originating from partial suggests that natural trans fatty acids should be
hydrogenation, including in the presence of TFAs distinguished from industrial ones, regarding
from other sources. The determination of the their impact on human health. One trans isomer
presence of PHOs is based on the evaluation of at a time should therefore be considered in
the content of mono-unsaturated TFAs, and nutritional studies. However, a high amount
other selected fatty acids. Food samples were combined with an excellent purity of the
prepared for gas chromatographic analysis corresponding isomer remains a challenge. This
according to AOCS Official Method Ce1k-09. The study aimed at providing means to overcome
gas chromatographic quantification of fatty acids, such issues.
including individual trans-18:1 positional isomers,
Methods Used: The purification of several grams
was achieved with an SLB-IL111 (200 m x 0.25 of trans-palmitoleic acid (C16:1 n-7 trans, TPA)
mm x 0.20 µm) capillary column applying the was considered in this study, starting from the
method developed by Delmonte et al (2011). The cis-palmitoleic acid (C16:1 n-7 cis, CPA). Fatty
accuracy of the method was tested by analyzing acids were used as ethyl esters. Several sources
various vegetable oils spiked with different of CPA were evaluated. The following means
quantities of PHOs. The method could accurately were critically assessed: Flash HPLC, urea
detect PHO in 96–97% of spiked samples complexation, and crystallization in solvent.
containing at least 0.5% TFAs (% of total fat). The Within each step, the purity was checked by GC-
method was applied to the analysis of about 100 MS, and the best means was chosen.
commercial food samples, part of which reported Results: Macadamia nut was deemed as a
PHOs among their ingredients. reliable source of CPA, the other cis-C16:1
References: Pierluigi Delmonte, Ali-Reza isomers being present in low amounts.
Fardin Kia, John K.G. Kramer, Magdi M. Mossoba, Purification of CPA was best carried out using
Len Sidisky, Jeanne I. Rader (2011). Journal of Flash HPLC. The isomerization of CPA to TPA was
Chromatography A, 1218: 545–554 easily done. Removing as much CPA as possible
from the CPA-TPA mix was best performed by
crystallization in solvent, ensuring a 99%-purity of
the final TPA product.
Conclusions: Complementary means were used
to get high amounts of highly purified-TPA. Not
only ethyl esters were suitable for purification
purposes, but they made the purity control by
GC-MS easy. Provided the availability of the
– 20 –
corresponding cis-isomer, our purification Normal Phase UV Compatible HPLC Separation

strategy can be applied to any trans of Hydroxylated and Non-hydroxylated Lipids
monounsaturated fatty acid. for Metabolic Flux Analysis Hari Kiran Kotapati*
and Philip D. Bates, The University of Southern
Determination of sn2-position Fatty Acid in Mississippi, USA
Long-chain Triglycerides(LCTs) and Medium- The major objective is to develop a normal
and Long-chain Triglycerides(MLCTs) with phase HPLC method to completely separate
Enzymatic Alcoholysis by GC-FID Wei Ting Ting, hydroxylated and non-hydroxylated
Wen Ming Cao, and Yuan Rong Jiang, Wilmar triacylglycerols (TAGs) and diacylglycerols (DAGs)
(Shanghai) Biotechnology Research & classes for online liquid scintillation counting
Development Center Co., Ltd., China
within 14C based metabolic flux experiments.
A new sn2-position fatty acid composition
The secondary objective was to have a solvent
analysis method was established in this research.
system compatible with detection of lipids at 210
The method was not only used in plant oils with
nm to track lipid mass with a non-destructive and
long-chain triglycerides (LCTs), such as sunflower
commonly available detector for use with
seed oil, rapeseed oil, but also in medium- and
fraction collecting. Different stationary phases,
long- chain triglycerides (MLCTs). 2-
for normal phase HPLC, including silica and
monoaclyglycerols (2-MAGs) were achieved from
modified silica supports were examined for our
enzymatic alcoholysis of triglycerides and
studies during the course of method
analyzed by GC-FID after derivation. 2-MAGs as
development. PVA coated silica support provided
detected targets were proved to be appeared
excellent separations between the hydroxylated
and retention time was confirmed, and 1(3)-MAG
TAG classes from non-hydroxylated DAGs that
was absent by GC-QTOF. The condition test was
were very difficult to separate via previously
developed in this research and the experiment
reported chromatographic techniques for
condition, such as enzyme amount, reaction
hydroxylated neutral lipids. This normal phase
temperature, reaction time and derivative
HPLC analysis technique was combined with β-
reagent volume were optimized by single factor
ram detector to perform continuous flow liquid
test. 2-C8-MAG, 2-C10-MAG,2-C12-MAG, 2-C14-
scintillation counting, which is important for
MAG, 2-C16-MAG and 2-C18-MAG were
rapid and efficient quantification of labeled lipids
determinated in MLCT by GC-FID. The results of
from radioactive based enzyme assays or in vivo
long chain triglycerides compared with porcine
metabolic flux experiments. We have successfully
pancreatic lipase method were in satisfactory.
performed chromatographic separations of non-
This method was competitive to analyze
labeled individual lipid classes by UV detection,
triglycerides contained medium chain fatty acids
and radioactive lipids extracted from
which could not be analyzed by porcine
[14C]acetate in vivo metabolic labeling
pancreatic lipase method, and it provided shorter
experiments in developing Arabidopsis thaliana
pretreat time and better repeatability due to
seeds. In this study, the samples collected from
detected targets changed and SPE or TLC
separation removed.
– 21 –
different time points during the course of a 1,3,5-trimethyl-2,4,6-tris(3,5-di-tert-butyl-4-

continuous 14C metabolic labeling experiment hydroxybenzyl)benzene in the ratio of 1: 2. An
were analyzed and the biosynthetic precursor- amount of 10g of the combined adsorbent was
product relationships of major lipid classes were used per 1L of oil. The Magnesium aluminum
identified. hydroxycarbonate compound was placed on top
of 1,3,5-trimethyl-2,4,6-tris(3,5-di-tert-butyl-4-
Mitigating the Deteriorating Effect of Biofuel
hydroxybenzyl)benzene in the column. 100 ml of
in Engine Oil Jerome D.A. Kpan*1, and Juergen
Krahl2, 1Technology Transfer Automotive Centre the oil sample run through the adsorbent bed at
of Coburg University of Applied Sciences and Arts, a temperature of 130°C at a time. Also the
Germany, Germany; 2Coburg University of combined adsorbents were also added to the
Applied Sciences and Arts; Ostwestfalen-Lippe sample and aged. The acid number and peroxide
University of Applied Sciences, Germany value were determined. Size exclusion
This study presents the feasibility of using the chromatography was used to analyse the
process of adsorption in mitigating the negative oligomers. The FTIR analysis showed that
impact of the use of biodiesel on the crankcase adsorption suppressed about 80% formation of
oil. This is done by suppressing the buildup acidic products. This is collaborated with the size
and/or selectively removing oligomers formed in exclusion chromatography and the total acid
the oil as a result of the degradation of biodiesel. number. This study has shown that adsorption is
Biodiesel blended with neat base oil (20% a potential useful tool for the suppression of
biogenic fuel and 80% neat base oil) were thermo oligomers formation. Therefore, if biodiesel will
oxidatively aged. The separation of the oligomers be a significant fuel of the future then preventing
was carried out through fixed bed columns made its role in short oil change interval by the process
of Magnesium aluminum hydroxycarbonate and of adsorption is a great achievement.
– 22 –
ANA 3.1/PCP 3a: Bioprocessing for New/Value-added Protein Utilization:

Digestibiity Issues/Analytical Measurements
Chairs: Sneh Bhandari, Merieux Nutrisciences, USA; Buddhi Lamsal, Iowa State University,
USA; and Bishnu Karki, South Dakota State University, USA
Matrix Effect on the in vitro Immunodetection PARV19. Overall, it is never enough to emphasize
of Food Allergens. Qinchun Rao, Xingyi Jiang, and that matrix effect on target analyte quantification
Behnam Keshavarz, Florida State University, USA is unignorable during food allergen detection
To protect the public health, the U.S. food because any false negative assay outcomes may
manufacturers have been required to label food induce potential or severe life-threatening
allergens or ingredients derived from eight major allergic reactions in consumers.
allergenic foods since 2006. Currently, the
presence of misbranding and/or undeclared food Protein Quality Evaluation in Protein Enhanced
allergenic residues is the No. 1 cause of food Formulations Including Those Based on Oilseed
recalls in the US. In order to (1) fight food fraud, Based Proteins Sneh Bhandari*, Merieux
(2) better comply with the food regulations, (3) Nutrisciences, USA
decrease the food recalls economic loss to the There’s growing evidence that high-protein
food industry, and (4) reduce the risk of food food choices do play a role in health and more
allergy, it is necessary to develop reliable and consumers are looking for high quality proteins
robust in vitro detection methods to prevent the from varied sources including those from plants.
occurrence of undeclared allergenic residues in Oilseeds protein are becoming a newly
foods. As the major fish allergen, parvalbumins recognized source of dietary proteins particularly
(PV) from mullet and salmon in two sample to meet growing needs of large segments of
models were used to elaborate the relationship word population. More and more new protein
between matrix effect, extractability of PVs, and products are becoming available from different
their thermostability during in vitro oilseeds for opportunity to incorporate in a
immunodetection. Matrix-induced thermal broader variety of foods to make nutritionally
instability of PV was mainly due to physical enhanced products. The accurate assessments of
(hydrophobic effect) and chemical (thiol-disulfide protein quantity and quality in newly available
interchange) interactions. Our results illustrate sources of the dietary proteins and the
that the addition of sodium dodecyl sulfate (SDS, formulations and product based on those has is
surfactant), β-mercaptoethanol (reducing agent) important. This evaluation has acquired
or ethylenediaminetetraacetic acid (EDTA, metal additional importance now because of current
chelator) during sample preparation could not new trends of the development of protein
only increase the extractability of PV but also enhanced products. Protein quality needs to be
enhance its immunodetection using two PV- evaluated to determine its percent daily value for
specific monoclonal antibodies. Our findings nutritional labeling in US and can be done by the
demonstrate an overdose on EDTA made PV use of PDCAAS in food meant for ages >1 year.
monomer Ca2+-free and led it undetectable by Formulation of a nutritionally incomplete protein
– 23 –
with an ingredient containing complementary the DH value obtained with the methodology and
protein can result in a product with complete TNBS method, used as reference method. Then
protein with improved PDCAAS value. FAO has experimental validation tests realized with others
proposed a new protein quality measure enzymes were analyzed by SE-HPLC and TNBS
digestible indispensable amino acid score; DIAAS and pH-Stat methods. Good quantification of DH
which is yet to receive a wider acceptance by value was observed (90% of validation tests)
regulatory agencies. compared to TNBS method. The developed
methodology is a powerful tool for monitoring
Simultaneous Quantification of Hydrolysis enzymatic proteolysis both for animal and
Degree, Protein and Mean Weight of Peptides vegetable proteins while minimizing time.
Released during Enzymatic Proteolysis. Sophie Moreover, it could be used for functionalities,
Beaubier1, Irina Ioannou1, Xavier Framboisier2, digestibility or bioactivities analysis of produced
Olivier Galet3, and Romain Kapel2, 1LRGP—UMR
hydrolysates.
CNRS 7274, France; 2Reaction and Process
Engineering Laboratory UMR-7274, France; 3Avril
Nutritional Evaluation of Modified Carinata
Group, France
Meals in Finfish. Tom Kasiga and Michael Brown,
Enzymatic proteolysis is an industrial process
Dept. of Natural Resource Management, South
used in a wide range of applications Dakota State University, USA
(improvement of functionalities, nutrition, The recommended inclusion of carinata
bioactive peptides production…). Study of this Brassica carinata meal (CM) in animal feeds is
process consists in kinetic follow-up of the currently ≤10%. However, CM use in fish feeds
protein conversion rate, the hydrolysates size has not been tested but will likely be limited by
and the hydrolysis degree. To determine these 3 high concentrations of fiber, glucosinolates (GLS)
parameters, three different analysis are required and sinapine. GLS and sinapine tolerance in
which can have drawbacks particularly for Hybrid Striped Bass Morone chrysops ♀ X M.
vegetable protein hydrolysis. The communication saxatilis ♂ (HSB) was tested using incremental
presents an original methodology to quantify amounts of cold-pressed carinata meal (CPCM).
simultaneously these three criteria by size- Inclusion of >2.71 µmoles of GLS and >0.31 mg of
exclusion chromatography (SE-HPLC). The sinapine/g (>10% CPCM) of diet reduced feed
approach is based on absorbance profiles and the intake, resulting in reduced fish growth. To
estimation of molar extinction coefficient of each reduce antinutrients, we processed CM by
point of this one from the mixture aminogram of aerobic conversion (AC) using fungi ssp. followed
hydrolysates. Peak area of protein eluted into by a single wash to produce aerobically
column dead volume informs on protein converted carinata meal (ACCM). In a Rainbow
conversion rate and the peptide signal permits to Trout Oncorhynchus mykiss (RBT) trial, we
determine size and DH of hydrolysates. As a first replaced up to 75% of FM in a low animal protein
step, the approach was tested on the hydrolysis (20%) diet containing FM as the only animal
of animal and vegetable proteins with Alcalase protein source. Replacements ≥ 50% FM (≥10%
2.4L. A corrective factor was determined for each ACCM) reduced fish growth. Due to low
substrate from the linear correlation between
– 24 –
utilization of ACCM by RBT, we used low (20%) ACCM or 30% WCM. Thus, ACCM can replace
but similar animal protein contents (10% FM and more FM in diets but in combination with animal
10% poultry by-product meal) and included up to meals (≥20%). Because ACCM was low in GLS and
30% ACCM or 30% double-washed carinata meal sinapine but high in fiber, current research is
(DWCM) in HSB diets. Growth of HSB fed the FM focused on reducing fiber in ACCM.
reference diet was similar to that of HSB fed 30%
– 25 –
ANA 4: Trace Contaminants, including Processing Contaminants

Chairs: Jessica K. Beekman, US Food and Drug Administration, USA; and Mark W. Collison,
Archer Daniels Midland Co., USA
Comparison of Analytical Methodologies for the analytical methodologies for the analysis of
Analysis of Bound MCPD and Glycidol in Edible bound 3-MCPD and glycidol in an attempt to
Oils and Infant Formula Jessica K. Leigh*1, Kaitlin confirm method performance. Analytical
Grassi2, Shaun MacMahon1, Jan Kuhlmann3, methodologies, including those used by the U.S.
Adam Becalski4, Greg Jaudzems5, and Fabien Food and Drug Administration, Health Canada,
Robert5, 1US Food and Drug Administration, USA; SGS Germany, and Nestle, for the quantitative
2U.S. Food and Drug Administration, United
analysis of bound 3-MPCD and glycidol in various
States; 3SGS Germany GmbH, Germany; 4Health
samples of oils and infant formulas will be
Canada, Canada; 5Nestle Quality Assurance
compared. In addition, data will be presented
Center, United States
comparing the results of the use of direct and
Process-induced chemical contaminants
indirect detection methods with the infant
3-monochloro-1,2-propanediol (3-MCPD) esters,
formula extraction procedure developed by the
2-monochloro-1,3-propanediol (2-MCPD) esters,
FDA. Finally, a summary of the quantitative
and glycidyl esters are formed in edible vegetable
results obtained using each analytical method
oils during the deodorization step of the refining
will be presented.
process. As they are considered potentially
carcinogenic and/or genotoxic, their presence in
Detection Limits and Challenges in Low Level
refined oils and foods may pose a potential
Analysis of MCPD and Glycidol using AOCS
health risk. For this reason, research efforts over Method Cd 29c-13. Mark W. Collison and
the last several years have focused on developing Kevin Adlaf, Archer Daniels Midland Co., USA
methodology for the extraction and quantitation AOCS Method Cd 29c-13, “2- and 3-MCPD
of these contaminants in oils, infant formula, and Fatty Acid Esters and Glycidol Fatty Acid Esters in
other complex food matrices in an effort to Edible Oils and Fats by GC/MS (Difference
estimate levels of exposure and monitor possible Method)”, is widely used in industry because of
mitigation approaches from adjusting processing its rapid analysis time and ability to be
parameters. In addition, proposed EU regulations automated. Proposed MCPD and glycidol limits in
for free and bound 3-MPCD and glycidol infant formula are far lower than those proposed
concentrations in infant formula and edible oils for oil consumed by adults. The current
highlight the need for accurate analytical presentation centers on questions of whether
methodologies. Currently, there is no standard indirect GCMS methods are sufficiently sensitive
infant formula reference material containing for analysis of oils intended for or extracted from
known MCPD and glycidyl ester concentrations; infant formula, especially AOCS Cd 29c-13, and
therefore, verifying the accuracy of analytical discusses some of the challenges to achieving
methods has been difficult. The work discussed in very low limits of detection with indirect GCMS
this presentation compares a number of methods.
– 26 –
Recent Status of EU-regulation on 3-MCPD and Modern Analytical Tools in MCPD and Glycidol
Glycidol in Oils/Fats, Infant Formulae and Analysis: Research and Routine Analysis
Analytical Solutions Available Jan Kuhlmann*, Perspectives Katerina Mastovska*1, Vojtech
SGS Germany GmbH, Germany Hrbek2, Beverly Belkova2, Barbara A. Mitchell3,
Glycidyl fatty acid esters (GE; “bound Urairat Koesukwiwat4, and Jana Hajslova2,
1Covance Food Solutions, United States;
glycidol”) as well as 3-Mono-chloro-propane-
2University of Chemistry and Technology, Czech
diol esters (MCPD-E, “bound 3-MCPD”) are
known to be undesired food borne contaminants Republic; 3Covance Labs, Inc., USA; 4Covance
that frequently occur together in refined edible Food Solutions, Singapore
oils and fats and foods made from these. Free Traditionally, the analysis of
MCPD might be generated more frequently in monochloropropanediols (MCPD) in foods has
compound foods in course of heat treatment. It been focused on the determination of free 3-
might also migrate from diverse contact MCPD in a limited number of target, mostly
materials into foods. 3-MCPD and glycidol are hydrolyzed products using derivatization
classified as carcinogens (category 2B and 2A followed by GC-MS analysis. The discovery of
respectively) Recently the European Committee MCPD esters (re)ignited the interest in the
(EC) is working on regulations for the maximum toxicity, formation, occurrence, potential
levels of free and bound 3-MCPD as well as mitigation, and naturally also analysis of free and
glycidyl fatty acid esters in powdered and liquid bound forms of 3-MCPD and related compounds
infant formulae and related products as well as in (2-MCPD and glycidol). From the analytical
oils and fats intended for direct human perspective, this brought significant challenges in
consumption or use as an ingredient in food. The terms of the myriad of target analytes (various
draft regulation focuses as well on vegetable oils esters), broader scope of matrices and
and fats destined for the production of baby food concentration levels of interest. There are two
and processed cereal-based food. Accordingly, basic analytical approaches to the MCPD/glycidol
there is an obvious need for analytical methods analysis: direct determination of all free and
that meet recent and future regulation. In this bound forms or indirect analysis after conversion
regard, a guideline seems to be helpful as during (hydrolysis) to MCPD/glycidol free forms. The
the last 10 years, when the occurrence of indirect (total) analysis is suitable for high-
significant amounts of MCPD and glycidyl fatty throughput routine analysis and compliance
acid esters in various food matrices was testing. Similarly to the traditional analysis, the
reported, an extraordinary broad variety of free forms undergo derivatization followed by
analytical methods for the determination of GC-MS or GC-MS/MS analysis, the latter
these compounds have been published and some providing more selectivity for low-level analysis
also validated. This presentation intends to focus of highly complex matrices, such as infant
on the status quo of the European regulations on formula. The direct analysis is more suitable for
3-MCPD and glycidol derivatives and the research applications because it can provide
availability of analytical methods therefore. information about the composition and relative
abundances of the various ester forms in the
– 27 –
investigated samples. LC-MS/MS has been mostly toxicity of 2-MCPD was then evaluated using
employed for this purpose but there are other human hepatic cell line, Hep-G2. LD50 of 2-MCPD
techniques that can provide more information was several times higher than 3-MCPD, whereas
without the need for analyte-specific method 102 times higher than glycidol, when evaluated
conditions, such as non-targeted, high-resolution the cytotoxicity by measuring rates of the living
MS. Also, the LC separation could be replaced by and dead cells by Cell counting kit-8 and
supercritical fluid chromatography (SFC), which Cytotoxicity LDH Assay Kit-WST (Dojindo
offers different separation selectivity and some Molecular Technologies, Inc., Kumamoto, Japan).
unique possibilities for the analysis of Cytotoxicity of 2-MCPD was thus estimated to be
MCPD/glycidol esters. the lowest among the three substances, which
was consistent to our previous observation using
Toxicity Evaluation of 2-MCPD and Estimation Chinese hamster V76 cells.
of Intestinal Absorption of the Monoesters.
Yomi Watanabe1, Naoki Kaze2, Kaeko Murota3, The Importance of Aligning Analytical Limits
Hirofumi Sato4, Yuri Osafune5, and Araki with Health-based Guidance Values: Process-
Masuyama5, 1Osaka Research Institute of formed Compounds Case Study Paul R. Hanlon*,
Industrial Science and Technology, Japan; Abbott Nutrition, USA
2Ueda Oils & Fats MFG., Japan; 3Kindai University,
Exposure to substances created by food
Japan; 4Osaka Municipal Technical Research processing has been a part of the human
Institute, Japan; 5Osaka Institute of Technology, experience since the advent of cooking with fire.
Japan While human exposure to these substances has
Monochloropropane diol (MCPD), first
always been a part of our history, it is only
reported in vegetable protein acid-hydrolysate,
through the development of analytical methods
has 2 isomers. The major one, 3-MCPD, is
that we have become aware of the extent of our
categorized as group 2B; substances that are
exposure to these substances. The continued
possibly carcinogenic to humans by International
development of sophisticated methodology has
Agency for Research on Cancer. More
created a critical need for an aligned approach
toxicological evaluation and estimation on
between the risk assessment and analytical
intestinal behavior are required for the other
method development for these substances. Both
isomer, 2-MCPD. We have previously reported
of these processes feed into the development of
that 2-MCPD is estimated to be absorbed in
regulatory limits for these substances, however,
intestine in a similar manner to 3-MCPD when
the direct interaction between risk assessment
evaluated in vitro using intestinal epithelial cell
and analytical method development is often not
membrane model. The two detected similarly in
considered. Misalignment of these processes can
blood serum after their administration to rat
lead to confusion about the relevance of
duodenum, and was estimated to reach to hepar
detection of these substances at concentrations
in vivo. When 2-MCPD monoester was
below the health-based guidance values (HBGVs).
administrated to rat duodenum, free MCPD was
For example, detection of substances at trace
detected in blood serum in a similar manner to
levels has led to hazard-based evaluations where
that from 3-MCPD monoester, as well. The
– 28 –
any level of exposure is assumed to be pathways were proven by labeling experiments.

dangerous. Creating a model where fit for In addition, also examples for adduct formation
purpose methods are developed aligning limits of of these toxicants with other food ingredients
detection with HBGVs has the opportunity to will be presented. In summary, it will be shown
improve risk management activities by focusing that lowering the amounts of undesirable
activities on the areas that have the highest compounds in combination with the
priority for consumer safety. This is especially maintenance of an overall aroma well accepted
important for substances like process-formed by the consumers is a challenging task, but
compounds where the diversity of substances mitigation strategies of the “bad guys” can be
results in an increased opportunity for sensitive advised after getting the knowledge of their
analytical methods to detect “new” compounds. formation pathways.
Healthy but also Flavorful Food: Mitigation MOSH/MOAH and Plasticizers: Status quo
Strategies for Food-borne Toxicants Combined of Analysis and Activities of the Authorities
with Sensory Properties Accepted by Consumers in the EU Jan Kuhlmann*, SGS Germany GmbH,
Michael Granvogl*, Technical University of Germany
Munich, Germany Mineral Oil Saturated Hydrocarbons (MOSH)
In the past, many studies and efforts have together with Mineral Oil Aromatic Hydrocarbons
been undertaken to elucidate the key odorants of (MOAH) and plasticisers are complex groups of
food and to identify formation pathways of the undesired minor components in foods. Due to
so-called “food-borne toxicants” with a their lipophilic nature and ubiquitous distribution
subsequent mitigation strategy. But, up to now, they do occur in a broad variety of foods and also
analytical approaches including the quantitation frequently in edible oils and fats. NGO’s and
of desirable aroma-active compounds in Authorities as for instance the EFSA CONTAM
combination with undesirable toxicologically panel started to focus on these new
relevant substances by sensitive methods are contaminants since toxicological studies raised
scarcely available. The lecture will present recent indications for the possibility of undesired health
studies, which were combining the analysis of impacts on consumers. MOSH/MOAH as well as
important aroma compounds and of selected plasticisers represent groups of related single
food-borne toxicants (e.g., acrylamide, acrolein, components. As these groups might contain
crotonaldehyde, styrene, etc.) formed during more than hundred or in case of MOSH and
food-processing, e.g., brewing of beer or deep- MOAH even thousands of individual components
frying of potato chips and donuts in different risk assessment and analysis are challenging and
edible oils. Odorants were identified by gas compromises have to be taken. This presentation
chromatography-olfactometry as well as GC-MS tries to identify the recent opinion of the
and quantitated by stable isotope dilution authorities in the EU, which action might be
analysis (SIDA). For the toxicants, new taken in future and which analytical solutions are
quantitation methods using stable isotopically available so far.
labeled standards were developed and formation
– 29 –
Immuno Magnetic Solid Phase Extraction Effect of the Composition and Structure of
Combined with Cleanup to Determine Aflatoxin Excipient Emulsion on the Bioaccessibility of
B1 in Vegetable Oils Hongshun Yang* and Xi Yu, Pesticide Residue in Agricultural Products Ruojie
National University of Singapore, Singapore Zhang1 (Honored Student Award Winner; Lipid
Aflatoxin B1 (AFB1) contamination is a serious Chemistry and Nutrition Award Winner;) D. Julian
problem in edible oil industry. It causes severe McClements1, Lili He1, Zipei Zhang1, Wenhao
health threat and economic loss. In this study, a Wu2, Yeonhwa Park3, and Baoshan Xing2,
1University of Massachusetts Amherst, USA;
novel approach, low temperature cleanup (LTC)
2Stockbridge School of Agriculture, University
combined with immuno magnetic solid phase
extraction (IMSPE), was proposed to quantify of Massachusetts Amherst, USA; 3Dept. of
Food Science, University of Massachusetts
AFB1 in vegetable oils. LTC assisted in freezing
Amherst, USA
out the interference from oil matrix while IMSPE
Oil-in-water emulsions, such as dressings,
helped preconcentrate the targeted analyte. In
dips, sauces, and creams, are commonly
order to carry out the IMSPE, anti-AFB1
co-ingested with fruits and vegetables. Previous
monocolonal antibody functionalized magnetic
studies have shown that co-ingestion of
nanoparticles were first synthesized. Extraction
emulsions with fruits and vegetables may
of oil samples was then carried out followed by
substantially increase the oral bioavailability of
fluorometric quantitative analysis. The proposed
lipophilic nutraceuticals present within these
method showed satisfactory efficiency and
natural products. However, many natural
reproducibility with recovery rates in the range of
products also contain potentially detrimental
79.6%-117.9% and relative standard deviation
bioactive agents that may be introduced during
below 11.48%. Correlation of the method was
crop production and storage, such as pesticides.
satisfactory as well with linear correlation
The purpose of the current study was therefore
coefficient being 0.9941. The method was
to examine the potential impact of co-ingestion
applied to analyze five different kinds of
of emulsions with natural produce on the
vegetable oils from Singapore local supermarket
bioaccessibility of a hydrophobic pesticide.
including canola, peanut, soybean, corn, and
In current study, the influence of co-ingestion
olive oil. No AFB1 was detected in any of the
of food emulsions with tomatoes on the
mentioned oil samples. Overall, the proposed
bioaccessibility of a model pesticide
method is accurate, sensitive, and reliable. It is
(chlorpyrifos) was studied. The results indicated
convenient and promising in detecting trace
that the bioaccessibility of chlorpyrifos (a highly
chemical contaminants in oils.
lipophilic pesticide) was shown to depend on the
– 30 –
type and amount of co-ingested lipids in a model micelles formed from lipid digestion was more
food emulsion. Pesticide bioaccessibility important than the rate of their formation.
increased as the lipid content of the emulsions Overall, these results suggest that emulsions
increased and was higher when the lipid phase might increase the bioavailability of undesirable
consisted of long chain rather than medium chain pesticides, which may increase their risks to
triacylglycerols. However, the bioaccessibility did human health. Moreover, the results suggest that
not depend strongly on lipid droplet size, food effects should be taken into account when
suggesting that the total quantity of mixed assessing the potential toxicity of pesticides.
– 31 –
ANA 5: Marine Oils and Other Products

Chairs: Cynthia Srigley USA; Adam Ismail USA
Oxidative Status and Nutrient Label Claim process flavour and a negative correlation
Accuracy of the Top 50 Selling Omega-3 between primary oxidation products and acidic.
Products in the US Adam Ismail*, Global This research is a first step towards the broader
Organization for EPA and DHA Omega-3s, USA objective of standardizing the sensory
Abstract not available. terminology related to marine oils.
Sensory Vocabulary for Marine Omega-3 Oils An Examination of Marine and Vegetable Oil
Wenche Emblem Larssen*¹, E. Monteleone², M. Oxidation Data from a Multi-Year, Third-Party
Hersleth3, ¹Møreforsking, Norway. ²Dept. of Database Anna A. De Boer1, Adam Ismail2, Keri
Agricultural Biotechnology, University of Marshall3, Gerard Bannenberg2, Kevin L. Yan1,
Florence, Italy, 3Nofima, Norway and William J. Rowe1, 1Nutrasource, Canada;
2Global Organization for EPA and DHA Omega-3s,
Even though the Omega-3 industry has strict
restrictions concerning the chemical quality of USA; 3DSM Nutritional Products, USA
marine oil, they lack a defined methodology and Fish oil dietary supplements remain popular
a vocabulary for evaluating the sensory quality. sources of eicosapentaenoic (EPA) and
This study was conducted to identify the sensory docosahexaenoic (DHA) omega-3 fatty acids.
descriptors of marine oils and organize them in a However, some studies have suggested that
sensory wheel and vocabulary for use as a tool in commercially available fish oil supplements are
quality assessment. Oil-samples from six of the excessively oxidized, impacting oil quality and
largest producers of omega-3 products in Norway safety. Thus, we assessed oxidation data from a
were collected and analyzed both sensory, by six third-party database of 1900+ globally-sourced
industry expert panels and one trained sensory fish oil samples, including peroxide values (PV), p-
panel, and chemically. Through a series of anisidine values (p-AV), TOTOX, and acid values
language sessions, a total of 184 aroma (odor by (AcV). Additionally, we compared fish oils to krill,
nose), flavor, taste and mouthfeel descriptors algal, sunflower, and extra-virgin olive oil. Fish oil
were generated. A sensory wheel based on 60 products were predominantly compliant with
selected descriptors grouped together in 21 voluntary industry limits: for PV, 13.9% exceeded
defined categories was created to form a 5 mEq O2/kg (2.2% exceeded 10); for AcV 2.1%
graphical presentation of the sensory vocabulary. exceeded 3 mg KOH/g, while for p-AV in
In addition, a selection of the oil samples was unflavored oils, 6.1% exceeded 20, (3.8%
also evaluated by a trained sensory panel using exceeded 30), and 8.8% exceeded TOTOX limits
descriptive analysis and correlations between (26). Further, the fish oil median PV was similar
sensory and chemical analysis was studied. This to algal and sunflower oil, 4.8-fold greater than
showed a positive correlation between primary krill oil, and 5.2-fold less than extra-virgin olive
and secondary oxidation products and sensory oil. The fish oil median p-AV differed non-
properties such as rancidity, chemical flavour and significantly between oils. Finally, the fish oil
– 32 –
median AcV was similar to algal and extra-virgin values compared to bubbling oxygen after 30
olive oil, was 3.4-fold greater than sunflower oil, days. However, for hoki oil, the thermal
and 11.9-fold less than krill oil. This research has conditions produce p-anisidine values similar to
provided new insight that retail fish oil products the bubbling oxygen method. This suggests that
predominantly meet regulatory guidelines, and the two oils do not follow the same trend for
are comparable in oxidative status to other oxidizing rate with the three oxidation methods.
commercially available dietary oils.
Compositional Analysis of Algal Biomass, an
Chemical Changes During the Acute Oxidations Emphasis of Unique Contribution of Algal Lipids
of Fish Oils Austin S. Phung1, Selina C. Wang1, Lieve Laurens*, National Renewable Energy
Adam Ismail2, Gerard Bannenberg2, and Ameer Laboratory, USA
Taha3, 1University of California-Davis, Olive Algal lipid yields are unmatched by any
Center, USA; 2Global Organization for EPA and terrestrial feedstock, which makes their use ideal
DHA Omega-3s, USA; 2University of California, for renewable fuels, chemicals and food and feed
Davis, USA ingredients. Using algae as fuel and bioproduct
Fish oil is one of the most popular dietary precursor feedstocks has proven challenging due
supplements sold on the market. However, it to the lack of complete characterization of the
oxidizes more rapidly due to the presence of biomass and lipids. There remain many lipid
polyunsaturated fatty acids (PUFAs). We components unidentified or difficult to study.
measured the oxidative status of two fish oils, This presentation will cover the state of
hoki and anchovy, at varying time points over a technology of algal biomass characterization to
30-day period. To do this, we replicated the date and the current challenges moving forward.
oxidative conditions used in a previously As a case study, the utilization of ultra-high-
published article that oxidized hoki oil with resolution, fourier transform ion cyclotron
bubbling oxygen for 30 days under standard resonance mass spectrometry (FT-ICR-MS) for
fluorescent lights at room temperature. In the identification and quantification of novel
addition, two other oxidation methods were components will be discussed. Using a
evaluated – heating the oils in an oven at 50°C combination of Kendrick mass sorting and
with or without bubbling oxygen. Chemical elemental characterization of molecular ions,
measurements such as peroxide and p-anisidine series of compounds can be identified and help
values were used to assess the oxidation and with the elucidation of unknown components in
develop oxidation curves of both oils over the algal lipids. The superior mass accuracy and
30-day period. There are two main objectives of resolving power of the FT-ICR-MS in combination
this study: (1) to establish a more standardized with existing, more traditional, chromatography
fish oil oxidation protocol; (2) to estimate the and mass spectrometry tools has proven to be
oxidation status of fish oil products on the necessary for creating a reference library of novel
market by fitting on our oxidation curves. compounds found in algae. The data collected for
Preliminary results for anchovy oil suggest that a number of different species illustrates a
thermal conditions produce higher p-anisidine dynamic lipid profile and class distribution as the
– 33 –
cells are transitioning from early to late stage of Analysis of Omega-3 Polyunsaturated Fatty
cultivation. Acids (PUFA) in Phospholipid Oils: A Design of
Experiment Approach for Method Optimization
Sterol Fingerprinting in Algae, a New Method for Cynthia Srigley and Isa C. Orr-Tokle, US Food and
a New Feedstock Stefanie Van Wychen, and Drug Administration, USA
Lieve Laurens*, National Renewable Energy Phospholipid oil dietary supplements, such as
Laboratory, USA those containing krill or herring roe oils, have
Improving biomass production and utilization become increasingly popular alternatives to fish
is imperative for commercializing a future algae- oil supplements due to the enhanced
based biorefinery. Sterols are high-value bioavailability of omega-3 polyunsaturated fatty
products that can be isolated from algal biomass acids (PUFA) that the phospholipid structure
and have not been studied before in the context reportedly provides. Conventional methods for
of a conversion pathway to biofuels and analyzing fish oils, which typically contain fatty
bioproducts and provide value as either acids in triglyceride or ethyl ester forms, have
nutraceuticals or as feedstocks for novel proven ineffective when used on the
surfactant production. Mass spectral analysis phospholipid oil matrix. Therefore, the objective
indicates that there are 11 sterol compounds in of this study was to optimize a method for
Chlorella and 16 in Scenedesmus. The accurately quantifying fatty acids in phospholipid
compounds we identified fall within two major oils. Six different analytical methods were initially
classes: 28-carbon 4-desmethylsterols and 29- evaluated for their effectiveness in converting
carbon 4-desmethylsterols. Within the 28-carbon the fatty acids of a krill oil test material into fatty
group, the spectra either more closely resemble acids methyl esters for subsequent analysis by
campesterol or ergosterol. Within the 29-carbon gas chromatography with flame ionization
group, the spectra could be grouped into detection. The three methods which yielded the
compounds with a MW of 484 (such as greatest amounts of total fatty acids were taken
stigmasterol, fucosterol, Δ5-avenasterol) and for further testing, upon which Official Method
those with a molecular weight of 486 (such as Ce 2c-11 of the American Oil Chemists’ Society
sitosterol or Δ7-stigmastenol). We present data (AOCS) was found to produce the highest yield. A
on the development and thorough validation of a design of experiment approach was then
new, and much simplified, method based on performed to evaluate the effects of strength of
direct acid transesterification for sterol reagents, presence and type of antioxidant,
characterization, derived from either steryl esters inclusion of acid hydrolysis procedure, and
or glycosides in algae and highlight the growth duration and temperature of heating reactions.
phase specific accumulation of this class of Results indicated that AOCS Official Method Ce
compounds. 2c-11 is the optimal procedure for accurately
– 34 –
quantifying fatty acids in phospholipid oil. This other traditional WAX column. The excellent
official method was then applied to the analysis peak shapes obtained for acidic compounds,
of commercial phospholipid oil dietary eliminate the need to use two separate GC
supplements to evaluate their fatty acid columns for the analysis of free fatty acids (FFAs)
compositions and to measure their and their esters in the same sample. In this work,
eicosapentaenoic acid (EPA; C20:5n-3) and we present a variety of applications on the
docosahexaenoic acid (DHA, C22:6n-3) content.
analysis of free fatty acids, fatty acid methyl
Evaluation of an Ultra Inert WAX-phase Column esters (FAMEs) and fatty acid ethyl esters (FAEEs)
for the Analysis of Fatty Acids and FAMEs using DB-FATWAX Ultra Inert; including
Gustavo Serrano Izaguire, Allen Vickers, Yun Zou, determination of volatile FFAs from dairy
and Daron Decker, Agilent, USA products, analysis of Omega 3 and Omega 6 per
GC Columns with Polyethylene Glycol (PEG) AOCS Ce 1b-89, and FFAs and FAMEs in complex
stationary phases are commonly used for mixtures with other organic acids.
analyzing compounds with polar functional
groups, and are well suited for food, flavor, and Trans-fat Determination by Gas
fragrances applications. A current challenge with Chromatography Vacuum Ultraviolet Detection
traditional PEG phases, however, is the need to Jonathan Smuts*1 and Barbara A. Mitchell2, 1VUV
Analytics, USA; 2Covance Labs, Inc., USA
incorporate functional groups, such as
Vacuum ultraviolet detection for gas
nitroterephthalic acid, to separate challenging
chromatography (GC-VUV) has been applied to
analytes like acidic organic compounds. These
the analysis of fatty acid methyl esters (FAME).
modifications, unfortunately, reduces column
Here we specifically apply the technology for the
lifetime, maximum operating temperatures and
quantitative determination of trans-fat in various
are prone to react with some active analytes.
food samples. Two GC columns are explored,
Continuing with our recent advances in Ultra
100% cyanopropyl and a PEG-based column.
Inert (UI) technology, we are introducing a new
Linearity is demonstrated and then used to
PEG UI phase, specifically designed for the
calculate relative response factors. These are
analysis of fatty acids in free and ester forms.
then applied to a variety of food samples: fish oil,
This new GC Column (DB-FATWAX Ultra Inert),
biscuit, baby formula, gummy vitamin and butter.
delivers superior inertness, better long-term
Finally, quantitative GC/VUV results are
thermal stability and greater sensitivity than any
compared to GC/FID.
– 35 –
ANA-P: Analytical Poster Session
1. Purification of Native Cyanogenic Glycosides method Ch 6-91. The powder X-ray diffraction
from Flaxseed Veronique J. Barthet* and Tao (XRD) measurements were carried out for SS,
Fan, Canadian Grain Commission, Canada FPM and FPM:HOSO blends at 0, 20, 50 and 100%
The four cyanogenic glycosides (linustatin, FPM concentrations. The XRD analysis was
neolinustatin, linamarin, and lotaustralin) were performed at room temperature in a Philips PW
purified from intact mature and germinated 1710 diffractometer (PANalytical, Almelo,
flaxseeds after extraction followed by a Netherlands), using Bragg–Brentano geometry
preparative HPLC step. The confirmation of the (θ:2θ). The FPM profile presented 44.05% β-
identification of the purified compound was done sitosterol, 26.77% stigmasterol, 23.56%
by GC-MS. A single preparation experiment campesterol, and 5.62% corresponded to
generated from 3 to 14 mg of these cyanogenic minority compounds. The SS, FPM and
glycosides. Individual cyanogenic compound was FPM:HOSO blends showed similar XRD profiles,
obtained in a pure and native form allowing the with very small differences in peaks
increase of analytical method accuracy, displacement. The degree of crystallinity of the
reliability, and sensitivity. blends decreased with the increase of HOSO. In
this way, the crystalline behaviour of
2. Crystalline Pattern of Phytosterols in High phytosterols was not changed when it was
Oleic Sunflower Oil for Food Applications applied as a food grade mixture in a lipid system
Mayanny G. Silva, Valéria S. Santos, Lisandro P. (FPM:HOSO), even if it is compared to a pure
Cardoso, Maria Helena A. Santana, and Ana Paula
phytosterol (SS).
B. Ribeiro, University of Campinas, Brazil
The incorporation of phytosterols in foods
3. Thermal Properties and Solid Profiles of
has been receiving an increasing attention,
Hardfats-Soybean Oil Blends for Formulation of
because they are considered functional Lipid Carriers Mayanny G. Silva and Ana Paula B.
compounds. The bioactivity of phytosterols Ribeiro, University of Campinas, Brazil
resides in the fact that they have molecular Soybean oil (SO) is a widely available low cost
structures very similar to cholesterol. Thus, a raw material with high nutritional value. To
competitive absorption mechanism in the increase its functionality for the formulation of
intestine is established between them, reducing different types of lipid carriers, its technological
blood cholesterol levels. The objective of this properties can be adjusted with the addition of
study was to evaluate the crystalline pattern of hardfats, which are low-cost industrial products
the analytical grade stigmasterol (SS) (99% purity) resulting from the total catalytic hydrogenation
comparing to a commercial food grade free of liquid oils. The present work evaluated binary
phytosterol mixture (FPM), as well as to blends (50:50 w/w) of hardfats from palm kernel
investigate the crystalline behavior of FPM into (PKO), palm (PO), soybean (SO), microalgae (MO)
high oleic sunflower oil (HOSO). The phytosterol and crambe oils (CR) with SO, in relation to
profile of the FPM was characterized by AOCS thermal behavior and solid fat content profiles
– 36 –
(SFC). Thermal analysis of the samples was quantitative analysis of the methyl ester of each
performed by differential scanning calorimetry CLA (FAME-CLA) was variably underestimated.
according to the AOCS method Cj 1–94. The SFC To address this recovery concern of the major
was determined by nuclear magnetic resonance, CLAs found in our samples, three different
according to the AOCS official method Cd 16b-93; methylation procedures were evaluated with
direct method, measurements in series, of 10 to each of four CLAs (c9/t11, c9/c11, t9/t11, and
70ºC with interval of 5ºC. Melting point (TM) was t10/c12) as their methyl ester standards (>96%,
obtained from the SFC data, considered as the Matreya). Treatments in methanol were: (A)
temperature corresponding to a solids content of sodium hydroxide/boron trifluoride (NaOH-BF3,
4 g/100 g. At 25ºC, all blends showed a SFC 100⁰C, AOCS, Ce 2-66), (B) 4% sulfuric acid
between 17.8 and 51.4 g/100 g. These values (H2SO4, 80⁰C, AOAC 965.49), and (C) 4%
decreased with the increase in temperature in a methanesulfonic acid (MSA, 60⁰C, 2h). An
non-linear fashion until the complete melting Agilent GC equipped with a Supelco SP2560
between 33.7 and 67ºC. All blends showed large column (100M x 0.25mm x 0.20μm film) was
crystallization peaks, indicating diversity of the used for FAME analysis. Results show that the
triacylglycerol composition. Multiple melting recovery of each FAME-CLA was significantly
peaks were also observed, indicating higher (p<0.001) when treated with MSA than
recrystallization in more stable polymorphic with either H2SO4 or BF3. For (A) and (B),
forms. Blends of hardfats and liquid oils, therefor, recovery of c9/c11 FAME treated with H2SO4
stand out as a potential and low-cost option was significantly greater than with NaOH-BF3
for the formulation of lipid carriers due to (p<0.001), whereas, recoveries of c9/t11 and
the possibility of wide variation in their t10/c12 FAMEs were significantly greater with
physical properties. NaOH-BF3 than with H2SO4 (p<0.001). No
significant difference was observed for t9/t11
4. Comparative Recovery Analysis of FAME with either (A) or (B) treatment. The
Conjugated Linoleic Acids (CLA) Following recovery data suggest that more accurate
Different Methylation Protocols Yiyi Li, Raad S. determination, via greater recoveries, of FAME-
Gitan, Deborah L. Chance, James K. Waters, CLAs can be achieved with the use of 4% MSA
and Thomas P. Mawhinney, University of
in methanol.
Missouri, USA
Conjugated linoleic acids (CLAs) include a
5. 1H-NMR Measurement of Polar Phenolic
group of positional isomers of octadecadienoic Compounds: Reliable Determination of the
acids (18:2) that are naturally occurring Geographical Origin of Olive Oils Torben
polyunsaturated fatty acids, synthesized in the Küchler* and Ole Winkelmann, Eurofins Analytik
rumen of animals, found in related meat and GmbH, Germany
dairy products, and enhanced in grass-fed The mislabelling of olive oil with respect to its
animals. Early in our investigations of these geographical origin is a frequently encountered
products, it became apparent that, depending on fraud. According to European Regulation (EU) No
the methylation method employed, the 29/2012 it is mandatory to declare the
– 37 –
geographical origin of an olive oil on the bottle, trans-C18:1 fatty acids usually rely on gas
but so far no generally accepted analytical chromatography (GC), with flame-ionization
method exists to verify this labelling. Among the detector (FID) and hydrogen as gas vector.
three main producing countries in the Because hydrogen results in good peak
Mediterranean area, namely Italy, Greece and resolution, it is unclear whether the use of mass
Spain, customers particularly value Italian olive spectroscopy (MS) and helium as gas vector
oil. Presumably, olive oil from Greece and Spain would yield worse outcomes. Therefore, this
is mislabelled and sold as Italian olive oil on a study aimed at showing the abilities of GC-MS in
regular basis, as Italian olive oil is traded at a combination with helium when dealing with
higher price. The 1H-NMR analysis of olive oil positional isomers of trans-C16:1 and trans-C18:1
after dilution in CDCl3 has been exhaustively fatty acids.
described in many scientific studies related to Methods used: GC-MS in combination with
geographical origin already. Due to the inherently helium was used. Both partially hydrogenated
low sensitivity of NMR, this approach provides fish oil (PHFO) and dairy fat, derivatized as fatty
mainly information about the fatty acid profile acid methyl esters (FAME), were considered
and little information about minor components because of differences of trans-C16:1 and trans-
present in the sample like polyphenols. In our C18:1 isomer distribution. To rule out the effect
experience, this data can only be used to of stationary phase, two different GC columns
discriminate olive oil on a regional, but not on a were used. Optimized conditions determined in
country level. By employing a simple extraction each case.
with acetonitrile, a drastic enrichment of Results: Using optimized conditions, GC-MS
polyphenols and other polar compounds can be and helium led to good resolution of trans-C16:1
achieved. 1H-NMR analysis of these extracts can and trans-C18:1 positional isomers, in respect
be used to classify olive oils according to with what can be obtained with hydrogen and
geographical origin on the country level with FID. This held true for both dairy products and
striking success. Employing a database of about PHFO for which differences in profiles were
1000 reference samples that were collected clearly highlighted. Both GC columns provided
during the last five years, we are able to verify similar resolutions.
the geographical origin of olive oils from Italy, Conclusion: GC-MS in combination with
Greece and Spain with a high rate of success helium is suitable for analysis of positional
(> 95%). isomers of trans-C16:1 and trans-C18:1 fatty
acids.
6. Using GC-MS and Helium to Resolve
Positional Isomers of trans-C16:1 and trans- 7. A Microscopy Study of the Structure of
C18:1 Fatty Acids Etienne Guillocheau*, Daniel Njangsa and Other Selected Seeds: Method
Catheline, Philippe Legrand, and Vincent Rioux, Development A Microscopy Study of the
Agrocampus-Ouest, France Structure of Njangsa and Other Selected Seeds:
Objective and hypothesis: Separation and Method Development. Benjamain M.
analysis of positional isomers of trans-C16:1 and Bougouneau1, Michael Moore2, Samuel A.
– 38 –
Besong1, and Alberta N A Aryee1, 1College of lipids in seed tissues. DiOC is a viable stain for
Ag. & Related Sciences, Department of Human identifying total lipids in hydrated fixed plant
Ecology, Delaware State University, USA; 2Optical seeds, while confocal microscopy is a viable
Center for Applied Research, Department of modality for localizing macro-molecules in
Physics and Engineering, Delaware State hydrated seed sections. Conventional paraffin
University, USA embedding techniques for processing seeds
Njangsa seed contain about twice as much oil
would strip away lipids, our method preserve
as soybean (45—67%), and high amount of the
lipids for microscopy analysis. This new method
conjugated linoleic acid: alpha eleostearic acid.
require minimal sample preparation protocols
Black-eyed peas, and bush and Kent mango
and analysis can be performed on a time scale of
kernels are valued for their nutritional quality
seconds.
and potential antioxidant properties,
respectively. Very little is known of their 8. The LC-UV Analysis of 16 Cannabinoids of
microstructure. Microscopy studies may reveal Interest in Commercially Available CBD Oils.
structure, packing, and composition of the Joseph D. Konschnik, Justin A. Steimling, and
cotyledon, which may provide information on the Ashlee M. Reese, Restek Corporation, USA
seed’s susceptibility to enzymatic attack, stability More than 100 cannabinoids have been
under various processing conditions and pre- isolated from cannabis in addition to the five
extraction techniques for efficient component most commonly tested: THC, THCA, CBD, CBDA,
recovery. After using a series of unsuccessful and CBN. While methods have been published
sectioning techniques, black-eyed pea, mango that show the separation of these major
kernels, and Njangsa seeds were soaked in cannabinoids, many do not take into account the
springwater to soften the embryonic tissue, fixed possibility of interference from other
in 4% paraformaldehyde and vacuumed for 24 h. cannabinoids that may be present. This is most
The fixed seeds were cut into 30-40 micron problematic in concentrates where minor
sections using a tissue chopper and placed on a cannabinoids can be enriched to detectable
small shallow dish with Calcofluor white (a stain levels that were not observed in the flower.
that labels cellulose). The fluorescence created Additionally, some terpenes have been shown to
by the Calcofour white stain was animated under absorb UV light at 228 nm, the wavelength
a 405-laser line on the confocal microscope, and cannabinoids are typically detected, which can
starch granules were strongly polarized. DiOC- result in an additional source of interference. In
18(3) stain marked proteins and lipoproteins this study, the LC-UV separation of 16
under a 488-laser line on the confocal cannabinoids of interest was performed while
microscope. Various features and cellular monitoring for the potential impact from minor
constituents, mostly small spherical protein, and cannabinoids and terpenes on reported potency
oil bodies, and lipoprotein interspersed within values. The method is applied to commercially
the cytoplasmic network were visualized. available CBD oils that have recently become
Swelling and fixing seed tissue prior to thin suspect due to inaccurate label claims.
sectioning is a feasible method for preserving the
– 39 –
9. Rapid Measuring and Modelling Total Polar source for mammalian infants, however, there is
Compounds in Frying Oils using a Flash Gas a limited understanding of the dynamics of fat
Chromatography Electronic Nose Lirong Xu1, digestion in these structures. In this study, we
Li Xu2, Qingzhe Jin3, and Xingguo Wang3, have developed an Electron Paramagnetic
1Jiangnan University, China; 2School of Food
Resonance (EPR) spectroscopy approach to
Science and Technology, Jiangnan University, evaluate real time dynamics of MFGs interfacial
China; 3Jiangnan University, China
structure during simulated intestinal digestion.
A flash gas chromatography electronic nose
To measure these dynamics, MFGs were labeled
(FGC E-nose) was used for rapid measuring total
with spin probes, partitioning of EPR probes into
polar compounds (TPC) in frying oil samples. TPC
MFG membrane was validated using saturation-
of frying oil samples was measured by the AOCS
recovery measurements to estimate the depth
official method for reference. Qualitative
parameter (Φ). After validation, the selected spin
discrimination between frying oil samples with
probe was used to evaluate MFG membrane
different frying time was conducted using the
fluidity as a measure of the interface’s integrity in
FGC E-nose in combination with discriminant
the presence of bile salts and pancreatic lipase.
function analysis (DFA). The different TPC values
Independently, bile salts were found to have a
in the samples were calculated by partial least
rigidifying effect on the spin probed monolayer of
squares regression (PLS) model from FGC E-nose.
the membrane, while pancreatic lipase resulted
The DFA result indicated that the FGC E-nose
in an increase in membrane fluidity. Collectively,
could be used for differentiation of frying oil
the effect of lipase appears to be diminished in
samples with different frying time. The results
the presence of bile salts. These results indicate
showed a good linear relationship (R2=0.98;
the efficacy of EPR in providing an insight
SD=1.433) between TPC in oil samples measured
into small time scale molecular dynamics of
by AOCS method and that estimated by PLS
phospholipid interfaces in MFGs. Understanding
model, indicating that the proposed approach
interfacial dynamics of naturally occurring
can be used as an alternative to AOCS method as
complex structures can significantly aid in
an innovative tool for rapid measurement of TPC
understanding the role of interfacial composition
in frying oils.
and structural complexity in delivery of nutrients
10. Electron Paramagnetic Resonance during digestion.
Spectroscopy Study of Milk Fat Globule
Membrane Dynamics during Simulated 11. Infrared Spectroscopy and PLS Procedures
Digestion Maha Alshehab, Madhu S. for the Rapid Prediction of EPA and DHA
Budamagunta, John C. Voss, and Nitin Nitin, Contents in Marine Oil Dietary Supplements
University of California, Davis, USA Sanjeewa R. Karunathilaka, Cynthia Srigley, Betsy
Lipid digestion is an interfacial process J. Yakes, Sung Hwan Choi, Lea Brückne1, and
Magdi Mossoba1, 1US Food and Drug
involving interactions of enzymes and bile salts
Administration, USA
with the lipid body interface. Milk Fat Globules
Long chain omega-3 (n-3) polyunsaturated
(MFGs) with their unique interfacial structure and
fatty acids are predominantly found in marine-
membrane composition are a key nutritional
– 40 –
derived foods and dietary supplements, including The United States Food and Drug
fish oil (FO) supplements. Due to the potential Administration (FDA) requires disclosure of trans
health benefits, FO dietary supplements are now fat content of conventional foods and dietary
the third most commonly used supplements next supplements. Sample preparation for the most
to multivitamin-mineral and calcium-containing common methods involves derivatization of the
supplements. The high demand for these FO hydrolyzed free fatty acids, converting them over
dietary supplements requires strict process as methyl esters (FAMEs) followed by high
control and quality assurance measures to resolution GC-FID analysis. Overall fatty acid
monitor FA concentrations, which in turn need composition can be determined with
simple, fast, and accurate analytical techniques Polyethylene Glycol (PEG) capillary columns
to verify product compositions. Infrared where double bonds of unsaturated fatty acids
spectroscopy (IR) is a highly desirable analytical are mainly of cis configuration. Separation and
technique for process control and monitoring differentiation of cis / trans structures require
applications, due to its simplicity, limited sample long length, highly polar columns containing
pre-treatment requirement, and rapid biscyanopropyl stationary phases. GC methods
measurement time. Two IR spectroscopic are quite slow; AOAC 996.06 requires over an
methods including Mid-Infrared spectroscopy hour of GC run time per sample not including
(MIR) and Near Infrared spectroscopy (NIR) oven cool down. This work examines FAMES
combined with chemometrics were evaluated to using a high-cyano containing capillary column by
rapidly predict the FA contents, especially AOCS Method Ce-1j-07, AOAC Method 996.06, as
eicosapentaenoic acid (EPA; C20:5n-3) and well as other applications applying the use of
docosahexaenoic acid (DHA; C22:6n-3) of FO shortened narrow bore columns, high carrier gas
supplements purchased from US-based online flows, and fast oven temperature programming
retailers. A broad-based single calibration set as routes to reduce GC run times. The trade-off
that consisted of representative samples from a between separation performance and analysis
variety of marine oil sources was used to develop time is explored with the conclusion that
partial least squares regression (PLSR) calibration relatively complex mixtures of FAMES can be
models. This simple, fast, and nondestructive separated with greatly reduced analysis time.
quantitative method has the potential to be used
for the rapid screening of marine oil products for 13. Buffer Optimization for Accelerated SDS
verifying label declarations and also in quality Depletion by Transmembrane Electrophoresis in
control and monitoring during production. Top-down Proteomic Workflows. Subin R.C.K.
Rajendran1, Khaldun Al Azzam2, Nicole
12. Applying High Speed Gas Chromatography Unterlander1, and Alan Doucette1, 1Dept. of
for the Speciation of Fats in Foods and Edible Chemistry, Dalhousie University, Canada;
2Al-Ghad International College for Applied
Oils. Joseph D. Konschnik, Colton Myers, Kristi
Sellers, and Scott Adams, RESTEK Corporation, Medical Sciences, Saudi Arabia
USA Top-down proteomics entails the analysis of
intact proteins by mass spectrometry (MS).
Detergents such as sodium dodecyl sulfate (SDS)
– 41 –
play essential roles in sample processing (eg to produce trans fatty acids (tFA). To date, there
protein isolation/extraction, cell lysis, is no study describing the identification and gas
maintaining solubility) ahead of MS. chromatographic separation of the 16 geometric
Unfortunately, SDS is detrimental to MS. While isomers of 18:4n-3. In this study we have isolated
several strategies are available for SDS depletion, all the 18:4n-3 geometric isomers by silver ion
they suffer multiple concerns including HPLC (Ag+-HPLC), using up to 6 chromatographic
incomplete SDS removal, inadvertent protein columns in series. Each geometric isomer was
loss, or involving multiple complex and/or time- partially reduced with hydrazine to determine
consuming steps. Recently, our group introduced the geometric configuration of its double bonds.
an automated device for SDS depletion termed We have determined the gas chromatographic
transmembrane electrophoresis (TME) (Kachuk, elution order of all 18:4n-3 geometric isomers
J. Proteome Res., 2016, 2634). With TME, SDS utilizing a 100 m x 0.25 mm 100%
migrates under influence of an electric field as poly(biscyanopropyl siloxane) capillary column
the protein remains confined in a sample cell maintained at the elution temperature of 180°C,
behind semipermeable membranes. We achieve and have investigated their occurrence in
high protein purity (>99%) and high yield (>95%) Ahiflower oil. The 4 mono-trans 18:4n-3 isomers
in a relatively short (60 min) time frame. The produced during the refining of Ahiflower oil
present study aims to improve SDS depletion were chromatographically resolved from each
through examination of the influence of sample other, but c6,t9,c12,c15-18:4 co-eluted with the
composition, ultrafiltration membrane, TME tetra-cis isomer. These two fatty acids were
buffer properties (pKa, conductivity) and applied resolved by reducing the separation temperature
electric field on TME. Tris-tricine buffer system to 150°C, but this change produced tetra-cis
allows application of higher currents with 18:4n-3 to co-elute with t6,c9,c12,c15-18:4.
reduced Joule heating, resulting in faster SDS Combining the results from two isothermal
depletion. TME therefore presents a rapid, single- separations (180°C, 150°C) was necessary to
step and robust tool for preprocessing samples quantify the 4 mono-trans 18:4n-3 fatty acids in
ahead of proteomic analysis. Ahiflower oil.
14. Isolation and Identification of Stearidonic 15. A Method for Analyzing TAGs Composition
Acid Geometric Isomers. Pierluigi Delmonte, of Human Milk Fat using UPC2-Q-TOF-MS.
Andrea Milani, U.S. Food and Drug Xinghe Zhang1 and Guanjun Tao2, 1School of Food
Administration, Center for Food Safety and Science and Technology, Jiangnan University,
Applied Nutrition, USA China; 2State Key Laboratory of Food Science and
Seed oils rich in 18:4n-3 (Stearidonic acid, Technology, School of Food Science and
SDA) are marketed as a more sustainable, cost Technology, Jiangnan University, China
effective alternative source of omega-3 fatty Triacylglycerols (TAGs) of human milk fat
acids compared to marine oils. The refining of (HMF) is one of the most complex lipids in
these oils is expected to cause the geometric nature. It’s difficult to identify their categories
isomerization of the highly unsaturated 18:4n-3 and determine their content. We analyzed TAGs
– 42 –
in HMF using Ultra Performance Convergence A simple configuration for this analysis connects
Chromatography (UPC2) coupled with quadruple headspace sampling techniques with GC or GC-
time-of-flight mass spectrometry (Q-TOF-MS). MS systems. This presentation will discuss this
UPC2 separates TAGs with highest efficiency. system and methodologies for the rapid and
While SURVEY method was used to collect robust analysis of terpenes in cannabis and
MS/MS spectrum signal automatically by cannabinoid samples. By optimizing flows and
selective bombardment of specific parent ions, temperatures in both the GC and Headspace
when MS spectrometry was carried on. Through system and changing to better columns the
this manner, different TAGs sharing the same methods are faster and more selective.
retention time and mass charge ratio (m/z) was
identified. The result indicated that 93 TAGs 17. HPTLC with Tandem MS and HR-MS for
types of human milk were identified. The USU Structural Identification in Lipidomic and Other
type of TAGs (unsaturated fatty acid at sn-1,3 and Complex Lipid Samples. Vicente L. Cebolla1,
saturated fatty acid at sn-2 position) accounted María P. Lapieza2, Luis Membrado1, Maria
Savirón3, Jesus Orduna4, and Judith Nichols*5,
for about 48.62%. There were 24 types of TAGs 1Instituto de Carboquimica/CSIC; 2Instituto de
show content more than 1%. OPL (15.58±0.04%)
Carboquimica/CSIC, Spain; 3CEQMA/CSIC, Spain;
was the predominant TAG in China human milk, 4ICMA / CSIC; 5CAMAG Scientific, Inc., USA
followed by OPO (13.43±0.05%), OOL We report that high-performance thin-layer
(8.30±0.01%), OLL (6.14±0.02%). And the results chromatography (HPTLC), a popular technique
show significant difference between individuals. for providing lipid-class separations, can also be
Some oil and fats, such as refined Bashar fish oil, useful for direct, rapid, and relevant structural
lard, walnut oil, olive oil, peanut oil and soybean identification of individual molecular species
oil were analyzed by this method, showed that within each lipid class. After HPTLC separation of
this method is suit for separating and analyzing lipid mixtures using gradient-based AMD2, the
TAGs from different sources. zones of interest on the plate can be selectively
on-line transferred to an ESI-MS instrument via
16. The Rapid Analysis of Terpenes in Cannabis. an elution-based interface. MS/MS (MSn, using
Ron R. Honnold, Agilent, USA an ion-trap instrument) and HR-MS spectra
There is a growing need for the analysis of (QToF) can be obtained because the respective
terpenes in cannabis and cannabinoid sodium adducts were fragmented in the positive
concentrates and we are collaborating with ion mode, and sodium remained the charge of
major laboratories to rapidly develop high-end their fragment ions. Likewise, semiquantitative /
instrument configurations and methodologies comparative profiles of individual molecular
that are robust, accurate and precise while still species in a lipid class can be obtained by
allowing for easy to implement workflows in high ESI-MS, as:
productivity laboratories. These methods must
assure safety and quality and the list of terpenes i. ionization efficiencies are similar for
that are to be analyzed is continuously being molecular species belonging to a given lipid
increased by state and local governing bodies. class;
– 43 –
ii. the development solvent is evaporated using FTIR-ATR and GC-MS analytical techniques.
before detection; The experimental set up consisted of two syringe
iii. the interface elution solvent has a constant pumps that injected controlled amounts of
composition and, in this way, similar degassed water and hemp seed oil through a
ionization efficiencies are obtained for the stainless-steel coil reactor pressurized by a
individual lipids in a class. This work focuses manual backpressure regulator and maintained
on profiling and identification of neutral lipids
at specific temperatures inside a temperature-
(NLs), sphingolipids (SLs) and phospholipids
controlled oven. After cooling in an ice bath,
(PLs), in three analytical cases:
samples were collected from the reactor exit
1) mono (MGs) and diacylglycerides (DGs) stream, centrifuged, and the oil layers used for
(in positive ion mode) and fatty acids analyses. FTIR-ATR analysis was done using 100
(FAs) (in negative ion mode) as impurities
repetitive scans to determine the absorbance
in a fatty acid methyl ester (FAME)-based
readings over 14 specific wavenumbers looking
biodiesel sample;
2) molecular species of neutral sphingolipids for probable different chemical functionalities.
(SL), such as sphingomyelins (SMs) and GC-MS analysis was performed to identify
globotriaosylceramides (Gb3), in human remaining fatty acids and their decomposition
plasma; products. FTIR-ATR analysis showed that
3) phosphatidylcholines (PCs), elevation of reaction temperatures favored the
phosphatidylethanolamines (PEs), and formation of aldehydes and fatty acid dimers as
phosphatidyglicerols (PGs) associated to judged by the intensity of absorbances at 1700
membrane proteins of photosynthetic and 900 cm (-1) wavenumbers respectively,
purple bacteria. which significantly increased above 200°C. This
was confirmed by the GC-MS analysis that based
18. Identification of Degradation Products after
on the retention time correlations and spectra
Subcritical Water Hydrolysis of Hemp Oil using
identified the probable presence of long-chain
GC-MS and FTIR-ATR. Andres F. Aldana Rico1,
Ruben O. Morawicki1, Jerry W. King2, Rohana aldehydes, such as cis,cis,cis-7, 10, 13-
Liyanage2, Chris Mazzanti1, Marco E. Sanjuan hexadecatrienal, 9, 12, 15-octadecatrienal, and
Mejia1, and Antonio J. Bula Silvera1, 1Universidad polymerization products like dimers of palmitic
del Norte, Colombia; 2Critical Fluid Symposia, USA (C16:0) and linoleic (C18:2) acids.
When water reaches the subcritical region,
changes in the dielectric constant and ionic 19. Analysis of Heavy Metal Concentrations
product improve its hydrolysis capabilities of and Human Exposure from Hemp Oils and Hemp
non-polar compounds such as lipids. However, if Products. Patricia Atkins and Sean Curran,
SPEX CertiPrep, USA
the applied temperatures or exposure times are
The cannabis industry has flooded the market
excessive, molecule decomposition due to
with new products in need of analysis with few
thermal degradation can occur. This research
regulations to govern the industry. Recently,
aims to identify byproduct of thermal
concerns have arisen about safety of this,
decomposition species of hemp seed oil
unregulated market. Sadly, a potentially
hydrolyzed under subcritical water conditions
– 44 –
significant group of contaminants has been chromatography can be complex. Due to the
largely ignored: toxic metals. Federally legal or uniqueness of each new column and their use for
quasi-legal hemp products (hempseed oil, hemp extended periods of time, minor differences in
extracts, CBD oil & extracts), those without the chromatography can occur. Specifically, there
THC, are widely available on the market. Such can be fluctuations in elution order and
products are also used as base oil for the addition resolution. These potential issues can affect
of cannabis & cannabinoid extracts. However, efficiency in obtaining results. To improve
due to a ban on cultivation in the US, virtually productivity and quality, we designed a practice
all the hemp used is imported from other to maintain similar retention times across GC
countries. Studies of other consumable systems using a feature called retention time
commodities (spices, teas, grains, etc.) locking. Because fatty acids profiles contain
exported from around the world have reported multiple peaks and often have temperature
widespread heavy metal contamination. ramps, perfect alignment of retention times is
Cannabis plants (hemp & recreational varieties) difficult. Analysis of the retention time profiles
are bio-accumulators of heavy metals. In the for 15 columns of varying time of use showed
processing of agricultural products, a large that each column had a unique pattern. However
amount of plant material is consumed to extract the flow rates required to match target retention
concentrates and oils, thereby increasing the risk times leveled off after the elution of 9c, 12c, 18:1
of heavy metal contamination. The scope of this linoleic methyl ester. Taking advantage of this
study was twofold; firstly, to analyze various legal tendency, a simple formula to update column
hemp products currently on the market and, flow rates was created using only two marker
secondly, to use these as a model for methods peaks. Whether a lab contains one GC used for
development for testing of restricted products. many different assays or many GC’s dedicated to
Samples were digested using microwave fatty acid analysis, this best practice allows the
digestion and analyzed by ICP-OES and ICP-MS to analyst to quickly adjust column flow rates to
determine the elemental composition of these achieve uniform retention times after
products and the concentration of potentially maintenance or installation of a new column.
toxic heavy metals. A large number of the tested
hemp products detected heavy metal 21. Analysis of Vitamin D and Previtamin D
contamination. in Food Products. Jinchuan Yang, Waters, USA
Vitamin Ds are essential human nutrients that
20. Fatty Acid Analysis with Applied Retention are responsible for increasing intestinal
Time Locking. Barbara A. Mitchell, Scott absorption of calcium, iron, magnesium,
Wejrowski*, Youa Herr, and Thomas Vennard, phosphate, and zinc. The two most common
Covance Labs, Inc., USA forms of vitamin D in the diet are vitamin D2
For labeling and health concerns, the analysis (ergocalciferol) and vitamin D3 (cholecalciferol).
of food products for saturated fat, trans fat, and Vitamin Ds can thermally interchanged to
total fatty acids is now routine. However, when previtamin D forms. The previtamin Ds are also
dealing with a wide variety of matrices, the bioactive, and should be counted as part of the
– 45 –
vitamin D content. Recently, the FDA required to convert the fatty acids to FAMEs. These
that vitamin D content must be declared on all derivatized oils were then analyzed on several on
food labels (1). The analyses of vitamin D in foods commercially available cyano phases, and
often measure the main forms (D2 and D3) only. evaluated for time- and cost-efficiency. The
The previtamin D forms are often not measured. unique selectivity of the ZB-FAME column
This could cause some error in the vitamin D presented in this work provided shorter run
content. Recently, we developed a LC-MS based times using a cost-effective 30-meter length,
method that can measure the vitamin Ds and the while also providing complete resolution of a
previtamin Ds. (2) This method has been applied 37-component FAMEs mixture. In addition to
to a number of foods. The performance of the these optimizations, we discuss the possibilities
analysis method will be discussed. Reference: of post-run bake out and column lifetime
1) Food labeling: Revision of the nutrition and improvement offered by the column’s high
supplement facts labels, US Federal Register Vol. upper temperature limit.
81, No. 103, May 27, 2016 2) J. Yang, G. Cleland,
K. Organtini, Analysis of vitamin D and previtamin 23. New Method for Fast and Straightforward
D in food products, Waters app note Determination of Oxidation Stability of Fats
720006064en, July 2017. and Oils. Carolin Edinger, Anton Paar ProveTec
GmbH, Germany
22. Unique GC Column Selectivity for Time The quality of fats and oils strongly depends
and Cost-efficient Separation of Complex on their oxidation stability. In this contribution a
cis/trans Fatty Acid Methyl Esters in Food. new method for evaluating the oxidation stability
Ramkumar Dhandapani, Phenomenex, USA of fats and oils by determining the induction
Testing of Fatty Acid Methyl Esters (FAMEs) period is introduced. Under accelerated
not only allows for authentication of oil conditions (elevated temperature and pure
products, but also serves as an indicator of any oxygen pressure) a sample of 5 mL/4 g is
adulteration. Complex FAMEs including cis/trans examined in a sealed stainless steel test
fatty acids, however, have historically been chamber. Typical conditions of the method are
challenging to separate in a reasonable period of temperatures between 80 °C–140 °C and an
time. Commercially available cyano phase GC initial oxygen pressure of 700 kPa. These
columns traditionally have had analysis times of conditions initiate a rapid oxidation process,
nearly an hour and utilized column lengths up to which is monitored by recording the pressure
100 meters, which presents a costly, time- until a predefined pressure drop. It was found
consuming analysis process. In this study, we that the elapsed time until the pressure drop is
present a unique cyano-based GC stationary directly related to the oxidation stability of the
phase, Zebron ZB-FAME, which is optimized to sample. Correlation and precision studies
improve performance for complex FAMEs demonstrate the method’s effectiveness. Due to
analysis. Samples of commercially available the defined oxygen volume in the closed test
canola oil, coconut oil, olive oil, and walnut oil chamber, the oxygen consumption can be
were derivatized using a BF3-methanol reagent calculated. Furthermore, we observed Arrhenius
– 46 –
behaviour with regard to the applied method was developed for compatibility of
temperature, enabling the user to determine the existing regulations at the time, which had most
activation energy of a specific oxidation process. solvent limits set at < 100 ppm. Current
An exemplary shelf-life investigation of pure and regulations now require a quantification range of
encapsulated flavor oils is presented showing the 1–5000 ppm. Multiple Headspace Extraction
impact of the optimum encapsulating system. (MHE) experiments were conducted on hemp oil,
The significantly reduced measurement time and and matrix interferences were evaluated over a
a high repeatability of the method represent range of concentrations. Linearity was evaluated
the major advantages of our method, allowing using multiple split ratios in order to cover the
for quick and direct measurement of the large concentration range required for analysis.
oxidation stability for research, process and Storage and handling stability of high-level
test bench control. standards was performed. Matrix interferences
were determined to be minimal. Linearity was
24. FET Analysis of Solvents in Cannabis Oil: acceptable over a range of 10–5000ppm, using
Adapting to Changing Regulations. Amanda FID detection and 1–5000ppm using MS
Rigdon1, Anne Jurek2, Julie Kowalski3, and detection. Solvent standard stability is acceptable
Frank Dorman4, 1Emerald Scientific, USA; 2EST over several uses.
Analytical, USA; 3Trace Analytics, USA;
4Pennsylvania State University, USA
25. Fast Simultaneous Determination of
Cannabis is a large and growing market, legal Capsaicin, Dihydrocapsaicin and Nonivamide for
in 28 states with $6B in revenue in 2017. A Adulteration in Edible and Crude Vegetable Oils
growing product in the industry is cannabis Coupled with UPLC-MS/MS. Chuan Zhou,
concentrates. In addition to simple vaporization Dianping Ma, Wen Ming Cao, Hai Ming Shi, and
by the end user, cannabis concentrates may be Yuan Rong Jiang, Wilmar Biotechnology Research
used to prepare alternate dosage forms such as & Development Center (Shanghai) Co., Ltd.,
tinctures, topical salves, and transdermal China, China
patches. Like all pharmaceutical products Capsaicinoids are pungent components in hot
potentially harmful residual solvents from the peppers, which have been detected in waste
manufacturing process must be minimized and cooking oil. However, trace analysis of
monitored. While dissolution methods for capsaicinoids in edible and crude vegetable oils is
residual solvents analyses exist (e.g., USP), a challenging task due to the complex matrix. In
complex matrices such as cannabis concentrates this study, a simple liquid-liquid extraction and
usually require matrix-matched standards to solid phase extraction (SPE) coupled with RP-
ensure accurate quantification. Through the use UPLC-ESI-MS/MS method was developed for the
of full evaporation technique-headspace gas quantification of capsaicinoids in edible and
chromatography, accurate quantification for crude vegetable oils to screen the adulteration
residual solvents can be achieved in concentrates with waste cooking oil. This method was used to
without the need for matrix-matched standards. simultaneously determine 3 capsaicinoids
Established in 2013, the industry-standard FET (capsaicin, dihydrocapsaicin, and
nordihydrocapsaicin) with capsaicin-d3, and
– 47 –
dihydrocapsaicin-d3 as internal standards. This limit of quantitation was decreased to 1.09-3.31

allows the complete analysis of a sample with μg/kg and recovery range from 73%-123%.
only an hour of time, even with sample Compared with previous methods, it has simple
pretreatment and chromatography. The linear pretreatment process and stable performance
range of 3 capsaicinoids ranged between 0.5 and that relative standard deviation was controlled
40 µg/kg. The limit of detection (LOD) and limit of under 3%. The methods on the four PAHs can be
quantification (LOQ) for capsaicinoids were satisfied almost of tocopherol products quality
calculated as 0.15 and 0.5 µg/kg, respectively. control requirements.
Quantitative recoveries ranging from 92.9% to
105% were obtained by the analysis of spiked oil. 27. A Primary Animal Fat Adulteration
The relative standard deviations were less than Application: Determination Branched Chain
5% (n=6). The established method could Fatty Acid in Beef and Mutton Tallow with GC-
potentially overcome the interference of Q-TOF & GC-FID and Evaluation. Tong Li, Peijin
Tong, Hong Yang, Wen Ming Cao, and Yuan Rong
triacylglycerols and fatty acids in edible and
Jiang, Wilmar Biotechnology R&D Center
crude vegetable oils and have been successfully
(Shanghai) Co., Ltd., China
applied to analyze real oil samples. This method Branched chain fatty acid (BCFA) is
provides a rapid and reliable method for the characterized fatty acid in ruminant animal oil,
detection of adulteration with waste cooking oils special for beef and mutton tallow. Methyl-12-
in vegetable oils. methyltetradecanoate, methyl-13-
methyltetradecanoate, methyl-14-
26. Determination Polycyclic Aromatic
methylhexadecanoate, and methyl-15-
Hydrocarbons in Tocopherol and Ether
Compound by Gas Chromatography Tandem methylhexadecanoate are typical branched chain
Mass Spectral. Tong Li, Ruifeng Zhang, Chuan fatty acid and widely distributes in beef mutton
Zhou, Hong Yang, Wen Ming Cao, and Yuan Rong tallow. it ultilize Methyl undecanoate as internal
Jiang, Wilmar Biotechnology Research & standard and was methyl-esterified with KOH-
Development Center (Shanghai) Co., Ltd., China menthol solution, and qualitied by GC-Q-TOF,
The novel methods have been developed and and then, determined by GC-FID. Meanwhile, the
confirm for determination of four polycyclic method has been compared three common GC
aromatic hydrocarbon (Chrysene, benzi(a)pyrene, column and evaluated separation of branched-
benz(a)anthracene, benzo(b)fluoranthene) in chain fatty acid, including SP-2560, from
tocopherol products, including mixed tocopherol, SUPELCO, HP-88 and CP-Sil 88, from Agilent. As
tocopheryl acetate, and tocopheryl succinate. For the result, SP-2560 has the best performance in
mixed tocopherol and tocopheryl acetate, they separation BCFA. This method preform well
were extracted by dimethylsulfoxide-methanol linearity and range from 5 mg/kg to 100 mg/kg,
solution. About tocopheryl succinate, it was and detected limitation was from 1–1.5 mg/kg;
purified by Molecularly Imprinted Polymers after quantity limitation range from 1.5–4 mg/kg. In
saponification. The methods have been applied beef tallow-palm oil binary blending system,
and validated in GC-QQQ with MRM mode. The using the signal BCFA as index to calculate
– 48 –
adulteration proportion, when ratio of beef 29. Extending GC Column and Detector Lifetime
tallow is more than 70%, the relative error can Using Cartridge Style Gas Management Filters
be lower than 10%. Ramkumar Dhandapani*, Phenomenex, USA
Gas chromatographic separation relays on
28. A Novel Method for Quantitative Analysis purity of carrier and detector gases. Impure gases
of Blend Oil Based on GC-FID and NPDA. can detector stabilization issues, baseline
Peijin Tong, Hong Yang, Wei Ting Ting, Tong Li, disturbance, peak tailing due to exposed active
Wen Ming Cao, and Yuan Rong Jiang, surface and column bleed. To prevent these
Wilmar Biotechnology R&D Center (Shanghai) troubleshooting issues from happening, cartridge
Co., Ltd., China style gas filters and gas traps were designed
Edible blend oil, which is mixed with a variety which were easy to install and easy to replace.
of oils according to the physiological metabolic Unlike traditional style gas traps, these filters
requirements, makes up for the deficiency of provided a sensitive color indicator that changes
nutritional function structure of single species color sharply indicating the saturation of
edible fat, as well as improving the stability and cartridge. To prove the significance of carrier gas
functional performance. Qualitative analysis of purity, experiments were performed in gas
edible blend oil is still a big challenge, not to chromatography with and without gas filters and
mention quantitative analysis. A novel method the results showed raised base line and excessive
for simultaneously identification and column bleed. Hence, the use of cartridge-based
quantification of edible blend oil based on novel gas management is proven to be essential part of
none preconditions database algorithm(NPDA) gas chromatography analysis to extend GC
was developed. The database of triacylglycerols column and GC detector's life time.
(TAGs) collected by GC-FID contains five common
oil species (614 samples in total). 20 groups of 30. Research on Nutrient Components of
binary and ternary blend oils were analyzed and Camellia Oil and Olive Oil in China Dong Zhang*,
processed separately, showing a robust result Lin Zhu, and Zhangqun Duan, Academy of State
with the accuracy of 88.14%. 15 binary and 10 Administration of Grain, China
ternary blind samples prepared were analyzed by China is the earliest country in the world
the developed method, and the accuracy was cultivating Camellia oleifera with a cultivating
92% with the relative errors less than 20%. This history of 2000-3000 years. In 1960s, Olea
method is based on a simple analysis to europea was imported and cultivated firstly in
determine the composition of TAGs with the China. The oil nutrient components of Camellia
most prominent advantage of quantifying the oleifera and Olea europea cultivated in China
contents of different type of oils in edible blend including fatty acid, triglyceride, tocopherol,
oil without knowing the composition of the oil squalene and sterol were investigated. The main
species in advance, showing a bright future in fatty acids in Camellia oleifera seed oil (camellia
quick segregation and quality control of edible oil) were oleic (74.31%–83.65%), palmitic
blend oil in routine analysis. (7.12%–9.57%), and linoleic acid (5.43%–9.83%).
The percentage of mono-unsaturated fatty acid
– 49 –
in camellia oil was 77.50%–84.44%, while

8.67%–12.04%, 5.58%–10.24% of saturated
and poly-unsaturated fatty acid. The major
triglycerides were OOO, SLO and LOO, of which
OOO content was the highest (81.23%). α-
Tocopherol was the main composition of total
tocopherol and presented a content of 90%
(116–749mg/kg). Squalene in camellia oil was
21–336 mg/kg. Sterol was 1099-2298mg/kg, of
which β-sitosterol and Δ7-stigmastenol were the
main composition. Oleic (65.85%-80.12%) and
linoleic acid (2.55%–17.22%) were the main fatty
acids in Olea europea oil (olive oil). Triglycerides
in olive oil mainly included OOL, POL+SLL, OOO,
SOL, POO, SOO, and POS+SSL. OOO and SOL were
52.77%–72.19%, 24.47%–40.18%, respectively.
The α-tocopherol content was 109.38–310.50
mg/kg and 2.43–33.98 mg/kg of γ-tocopherol.
Squalene in olive oil was 1913.04–7042.63
mg/kg. Sterol was above 1000 mg/kg in olive oil,
of which β-sitosterol with 84.52%–87.08% and
Δ5-avenasterol with 6.09%–8.23% were the main
component.
– 50 –

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ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018

ANA 1b: Lipidomic Analysis

Comprehensive Dual Liquid Chromatography Development of Lipidomics-based Reference

ANA 2b: Olive Oil, including Sensory Analysis

ANA 2d/LOQ 2b: Sensory Analytics and Analytical Methods

ANA 3: General Analytical

corresponding cis-isomer, our purification Normal Phase UV Compatible HPLC Separation

different time points during the course of a 1,3,5-trimethyl-2,4,6-tris(3,5-di-tert-butyl-4-

ANA 3.1/PCP 3a: Bioprocessing for New/Value-added Protein Utilization:

ANA 4: Trace Contaminants, including Processing Contaminants

any level of exposure is assumed to be pathways were proven by labeling experiments.

ANA 5: Marine Oils and Other Products

ANA-P: Analytical Poster Session

dihydrocapsaicin-d3 as internal standards. This limit of quantitation was decreased to 1.09-3.31

in camellia oil was 77.50%–84.44%, while

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