ANA 1A: Spectroscopic, Spectrometric and Chemometric Methods for Lipid Analysis Chairs: Sanjeewa Karunathilaka, US Food and Drug Administration, USA; and Bernd W.K

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

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

– 2 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 2 1 1 Hoeven , and Dominik Burger , DSM Nutritional essential to maintain process efficiency. Various 2 Products, USA; DSM 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

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

– 7 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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; Chromatography Coupled with High Resolution 2University of North Carolina at Charlotte, USA 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; 2 3 the fact that the quality of this product on the Eurofins Analytik GmbH, Germany; Institute 4 market has reached high standards. The current of Agriculture and Tourism, Croatia; Institut 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

– 9 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 2 Laura Del Coco , Federica Angilè , and Francesco Coco, L., Fanizzi F.P. (2016) Eur J Lipid Sci Tech 2 1 Paolo Fanizzi* , Department 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; and references therein 2Dipartimento di Scienze e Tecnologie Biologiche 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

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

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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 Chickpea Germination Improves the crackers, are a top saturated fat contributor 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018

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), 2 3 extract (OSGT) were tested. Comparing one-year Spain; Instituto de la Grasa (CSIC); University 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018

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 31 P 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 31 monitored by P 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 31 PG, PA) were found and determined by P 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 minor compounds. Tocopherols are typically Containing Vegetable Oils Sneh Bhandari , 1 2 considered to occur in their free form in foods. Ming Gao , and Pierluigi Delmonte , 1 2 Recently, we noted the presence of tocopheryl Merieux Nutrisciences, USA; US Food 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018

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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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, 1 1 2 Beaubier , Irina Ioannou , Xavier Framboisier , 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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%

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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, 5 1 Robert , US Food and Drug Administration, USA; SGS Germany, and Nestle, for the quantitative 2 U.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.

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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, glycidol”) as well as 3-Mono-chloro-propane- 1Covance Food Solutions, United States; diol esters (MCPD-E, “bound 3-MCPD”) are 2University of Chemistry and Technology, Czech 3 4 known to be undesired food borne contaminants Republic; Covance Labs, Inc., USA; Covance 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 4 Japan; Osaka Municipal Technical Research processing has been a part of the human 5 Institute, Japan; Osaka 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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

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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, 1 novel approach, low temperature cleanup (LTC) University of Massachusetts Amherst, USA; 2 combined with immuno magnetic solid phase Stockbridge School of Agriculture, University 3 extraction (IMSPE), was proposed to quantify of Massachusetts Amherst, USA; Dept. 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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.

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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 1 2 Hersleth3, ¹Møreforsking, Norway. ²Dept. of Database Anna A. De Boer , Adam Ismail , Keri 3 2 1 Agricultural Biotechnology, University of Marshall , Gerard Bannenberg , Kevin L. Yan , 1 1 Florence, Italy, 3Nofima, Norway and William J. Rowe , Nutrasource, Canada; 2 Even though the Omega-3 industry has strict Global Organization for EPA and DHA Omega-3s, 3 restrictions concerning the chemical quality of USA; DSM 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 2 1 traditional PEG phases, however, is the need to Jonathan Smuts* and Barbara A. Mitchell , VUV 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.

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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

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

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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

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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 2 3 3 Li Xu , Qingzhe Jin , and Xingguo Wang , have developed an Electron Paramagnetic 1 2 Jiangnan University, China; School 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 1 Lipid digestion is an interfacial process J. Yakes, Sung Hwan Choi, Lea Brückne , 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 1 1 12. Applying High Speed Gas Chromatography Unterlander , and Alan Doucette , Dept. of for the Speciation of Fats in Foods and Edible Chemistry, Dalhousie University, Canada; 2 Oils. Joseph D. Konschnik, Colton Myers, Kristi Al-Ghad International College for Applied 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 type of TAGs (unsaturated fatty acid at sn-1,3 and Complex Lipid Samples. Vicente L. Cebolla , 2 1 saturated fatty acid at sn-2 position) accounted María P. Lapieza , Luis Membrado , 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;

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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, 1 1 Ruben O. Morawicki1, Jerry W. King2, Rohana aldehydes, such as cis,cis,cis-7, 0, 3- 1 1 Liyanage2, Chris Mazzanti1, Marco E. Sanjuan hexadecatrienal, 9, 2, 5-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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 – ABSTRACTS 2018 AOCS ANNUAL MEETING AND EXPO May 6–9, 2018 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 1 2 3 Rigdon , Anne Jurek , Julie Kowalski , and detection. Solvent standard stability is acceptable 4 1 2 Frank Dorman , Emerald Scientific, USA; EST over several uses. Analytical, USA; 3Trace Analytics, USA; 4 Pennsylvania 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

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

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