Applicability of MTBE-based lipid extraction methods assisted by microwave in food analysis. Statistical comparison and greenness evaluation with Soxhlet and Matyash. - 2026
Applicability of MTBE-based lipid extraction methods assisted by microwave in food analysis. Statistical comparison and greenness evaluation with Soxhlet and Matyash.
[en] This research investigates the applicability of a microwave-assisted extraction (MAE) methodology using methyl-tert-butyl ether (MTBE) as a one-step organic solvent extraction or in mixture with methanol and water compared to conventional extractions for lipid analysis in food samples. MAE methods were compared in terms of extraction yields and fatty acid (FA) composition to Soxhlet (using n-hexane, MTBE, and diethyl ether) and Matyash (MTBE/CH3OH/H2O, 10/3/2.5, v/v/v), considered gold-standard methodologies for lipid extraction. FAs, derivatized into methyl esters (MEs), were analyzed through a gas chromatography-flame ionization detector. One-way ANOVA and post-hoc Tukey tests were used to identify statistical differences in yields and FAMEs composition, resulting in <10 % with a p < 0.05. Methods using MTBE/CH3OH/H2O showed higher yields but also higher FAMEs variability compared to one-step solvent extraction methods. The extraction techniques showed comparable performance in terms of FAMEs composition. The greenness of the extractions were also evaluated using the AGREEprep metric.
Disciplines :
Chemistry
Author, co-author :
Bellinghieri, Carlo ; Université de Liège - ULiège > TERRA Research Centre > Chemistry for Sustainable Food and Environmental Systems (CSFES)
Giacoppo, Giulia; Department of Chemical, Pharmaceutical, and Agricultural Sciences, Via Luigi Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Purcaro, Giorgia ; Université de Liège - ULiège > Département GxABT > Chemistry for Sustainable Food and Environmental Systems (CSFES)
Cavazzini, Alberto; Department of Chemical, Pharmaceutical, and Agricultural Sciences, Via Luigi Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Pasti, Luisa; Department of Environmental and Prevention Sciences, Via L. Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Stevanin, Claudia; Department of Environmental and Prevention Sciences, Via L. Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Chenet, Tatiana; Department of Environmental and Prevention Sciences, Via L. Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Marchetti, Nicola; Department of Chemical, Pharmaceutical, and Agricultural Sciences, Via Luigi Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Maietti, Annalisa; Department of Chemical, Pharmaceutical, and Agricultural Sciences, Via Luigi Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Franchina, Flavio ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique, organique et biologique ; Department of Chemical, Pharmaceutical, and Agricultural Sciences, Via Luigi Borsari 46, 44121, University of Ferrara, Ferrara, Italy
Beccaria, Marco ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique, organique et biologique
Applicability of MTBE-based lipid extraction methods assisted by microwave in food analysis. Statistical comparison and greenness evaluation with Soxhlet and Matyash.
E.G. Bligh, W.J. Dyer, A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATION, Can J Biochem Physiol 37 (1959) 911-917. https://doi.org/10.1139/o59-099.
J. Folch, M. Lees, G.H.S. Stanley, A SIMPLE METHOD FOR THE ISOLATION AND PURIFICATION OF TOTAL LIPIDES FROM ANIMAL TISSUES, Journal of Biological Chemistry 226 (1957) 497-509. https://doi.org/10.1016/S0021-9258(18)64849-5.
V. Matyash, G. Liebisch, T. V. Kurzchalia, A. Shevchenko, D. Schwudke, Lipid extraction by methyl-terf-butyl ether for high-throughput lipidomics, J Lipid Res 49 (2008) 1137-1146. https://doi.org/10.1194/jlr.D700041-JLR200.
M. Pilecky, M.J. Kainz, L.I. Wassenaar, Evaluation of lipid extraction methods for fatty acid quantification and compound-specific δ13C and δ2Hn analyses, Anal Biochem 687 (2024) 115455. https://doi.org/10.1016/j.ab.2023.115455.
C.Z. Ulmer, C.M. Jones, R.A. Yost, T.J. Garrett, J.A. Bowden, Optimization of Folch, Bligh-Dyer, and Matyash sample-to-extraction solvent ratios for human plasma-based lipidomics studies, Anal Chim Acta 1037 (2018) 351-357. https://doi.org/10.1016/J.ACA.2018.08.004.
A. Arendowski, E. Sibinska, W. Mista, P. Fijalkowski, M. Zloch, D. Gabrys, P. Pomastowski, Study of sample preparation influence on bacterial lipids profile in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, Lipids 59 (2024) 13-26. https://doi.org/10.1002/LIPD.12383.
M. Gorgich, T.M. Mata, A.A. Martins, M. Branco-Vieira, N.S. Caetano, Comparison of different lipid extraction procedures applied to three microalgal species, Energy Reports 6 (2020) 477-482. https://doi.org/10.1016/J.EGYR.2019.09.011.
L. Merquiol, G. Romano, A. Cutignano, I. D’Ambra, Chemical markers in marine food web: A simple workflow based on methyl tert-butyl ether extraction for fatty acids and stable isotopes assessment in plankton samples, Mar Environ Res 190 (2023) 106087. https://doi.org/10.1016/J.MARENVRES.2023.106087.
F. Gutmann, R. Fritsche-Guenther, D.B. Dias, J.A. Kirwan, Comparing the Extraction Performance in Mouse Plasma of Different Biphasic Methods for Polar and Nonpolar Compounds, J Proteome Res 23 (2024) 2961-2969. https://doi.org/10.1021/ACS.JPROTEOME.3C00596/ASSET/IMAGES/LARGE/PR3C00596_0003.JPEG.
C. Claassen, J. Kuballa, S. Rohn, Polar Lipids in Starch-Rich Commodities to be Analyzed with LC-MS-Based Metabolomics-Optimization of Ionization Parameters and High-Throughput Extraction Protocols, Metabolites 2019, Vol. 9, Page 167 9 (2019) 167. https://doi.org/10.3390/METABO9080167.
A.C.R. Silva, C.C. da Silva, R. Garrett, C.M. Rezende, Comprehensive lipid analysis of green Arabica coffee beans by LC-HRMS/MS, Food Research International 137 (2020) 109727. https://doi.org/10.1016/J.FOODRES.2020.109727.
L. Lan, H. Zhou, R. Jiang, Y. Cao, J. Yuan, X. Mao, K. Wang, S. Ji, Q. Hu, Exploring lipid profiles and unveiling potential lipid markers in raw and vinegar-processed Schisandra chinensis fruit, Food Chem X 29 (2025) 102639. https://doi.org/10.1016/J.FOCHX.2025.102639.
D. Ferrara, M. Beccaria, C.E. Cordero, G. Purcaro, Microwave-assisted extraction in closed vessel in food analysis, J Sep Sci 46 (2023) 2300390. https://doi.org/10.1002/JSSC.202300390.
Y.M. Dai, K.T. Chen, C.C. Chen, Study of the microwave lipid extraction from microalgae for biodiesel production, Chemical Engineering Journal 250 (2014) 267-273. https://doi.org/10.1016/J.CEJ.2014.04.031.
A. Gonzalez-Arrojo, A. Soldado, F. Vicente, B. de la Roza-Delgado, Microwave-Assisted Methodology Feasibility for One-Step Extraction and Transmethylation of Fatty Acids in Milk for GC-Mass Spectrometry, Food Anal Methods 8 (2015) 2250-2260. https://doi.org/10.1007/S12161-015-0108-8/TABLES/8.
N.P. Brunton, C. Mason, M.J. Collins, Rapid microwave assisted preparation of fatty acid methyl esters for the analysis of fatty acid profiles in foods, Journal of Analytical Chemistry 70 (2015) 1218-1224. https://doi.org/10.1134/S1061934815100044/METRICS.
A. Fina, S. Mascrez, M. Beccaria, C. De Luca, J. Aspromonte, C. Cordero, G. Purcaro, A high throughput method for fatty acid profiling using simultaneous microwave-assisted extraction and derivatization followed by reversed fill/flush flow modulation comprehensive multidimensional gas chromatography, Advances in Sample Preparation 4 (2022) 100039. https://doi.org/10.1016/J.SAMPRE.2022.100039.
D. Ferrara, M. Beccaria, C.E. Cordero, G. Purcaro, Comprehensive comparison of fatty acid methyl ester profile in different food matrices using microwave-assisted extraction and derivatization methods and comprehensive two-dimensional gas chromatography coupled with flame ionization detection, Advances in Sample Preparation 11 (2024) 100124. https://doi.org/10.1016/J.SAMPRE.2024.100124.
M. Campaniello, V. Nardelli, R. Zianni, M. Tomaiuolo, O. Miedico, M. Iammarino, A. Mentana, Microwave-Assisted Extraction/UHPLC-Q-Orbitrap-MS-Based Lipidomic Workflow for Comprehensive Study of Lipids in Soft Cheese, Foods 13 (2024) 1033. https://doi.org/10.3390/FOODS13071033/S1.
J.S. Wilson, G. Petino, D.C. Knudsen, Geographic context of the Green Pistachio of Bronte, a protected designation of origin product, J Maps 14 (2018) 144-150. https://doi.org/10.1080/17445647.2018.1438318.
A. Schincaglia, L. Pasti, A. Cavazzini, G. Purcaro, M. Beccaria, Optimization of headspace high-capacity tool coupled to two-dimensional gas chromatography-mass spectrometry for mapping the volatile organic compounds of raw pistachios. A proof-of-concept on the classification ability by geographic origin, Food Chem 460 (2024) 140702. https://doi.org/10.1016/J.FOODCHEM.2024.140702.
A. Schincaglia, L. Pasti, A. Cavazzini, G. Purcaro, M. Beccaria, Optimization and Validation of a Cheaper, Safer, and More Sustainable Methodology for Aflatoxins Determination in Rich-Lipidic Matrices (Pistachio Nuts) Using Deep Eutectic Solvent Extraction and UHPLC-FLD Analysis, J Agric Food Chem 72 (2024) 20670-20678. https://doi.org/10.1021/acs.jafc.4c05094.
P. Hernandez-Alonso, M. Bullo, J. Salas-Salvado, Pistachios for health: What do we know about this multifaceted nut?, Nutr Today 51 (2016) 133-138. https://doi.org/10.1097/NT.0000000000000160.
M.A. Lopez-Bascon, M.D. Luque de Castro, Soxhlet Extraction, in: Liquid-Phase Extraction, Elsevier, 2020: pp. 327-354. https://doi.org/10.1016/B978-0-12-816911-7.00011-6.
M. Beccaria, Y. Zou, P.H. Stefanuto, A.L.M. Siqueira, A. Maniquet, M. Piparo, P. Giusti, G. Purcaro, J.F. Focant, Deeper investigation of oxygen-containing compounds in oleaginous feedstock (animal fat) by preparative column chromatography and comprehensive two-dimensional gas chromatography coupled with high-resolution time-of-flight mass spectrometry, Talanta 238 (2022) 123019. https://doi.org/10.1016/J.TALANTA.2021.123019.
D. Ferrara, M. Cescon, G. Giacoppo, V. Costa, G. Purcaro, N.D. Spadafora, C. Cordero, L. Pasti, A. Cavazzini, M. Beccaria, Composition and nutritional values of fatty acids in marine organisms by one-step microwave-assisted extraction/derivatization and comprehensive two-dimensional gas chromatography -flame ionization detector, Journal of Chromatography B 1236 (2024) 124074. https://doi.org/10.1016/J.JCHROMB.2024.124074.
F. Pena-Pereira, M. Tobiszewski, W. Wojnowski, E. Psillakis, A Tutorial on AGREEprep an Analytical Greenness Metric for Sample Preparation, Advances in Sample Preparation 3 (2022) 100025. https://doi.org/10.1016/J.SAMPRE.2022.100025.
A. Schincaglia, J. Aspromonte, F.A. Franchina, T. Chenet, L. Pasti, A. Cavazzini, G. Purcaro, M. Beccaria, Current Developments of Analytical Methodologies for Aflatoxins’ Determination in Food during the Last Decade (2013-2022), with a Particular Focus on Nuts and Nut Products, Foods 2023, Vol. 12, Page 527 12 (2023) 527. https://doi.org/10.3390/FOODS12030527.
American Oil Chemists’ Society, AOCS Official Method Am 2-93 - Determination of oil content of oilseeds, (2011).
C.L. Teo, A. Idris, Enhancing the various solvent extraction method via microwave irradiation for extraction of lipids from marine microalgae in biodiesel production, Bioresour Technol 171 (2014) 477-481. https://doi.org/10.1016/J.BIORTECH.2014.08.024.
The Commission of the European Communities, Commission Regulation (EEC) No 2568/91 of 11 July 1991 on the characteristics of olive oil and olive-residue oil and on the relevant methods of analysis, Official Journal of the European Communities (1991) L248/1-L248/83.
M. Beccaria, E. Moret, G. Purcaro, L. Pizzale, A. Cotroneo, P. Dugo, L. Mondello, L.S. Conte, Reliability of the ΔECN42 limit and global method for extra virgin olive oil purity assessment using different analytical approaches, Food Chem 190 (2016) 216-225. https://doi.org/10.1016/J.FOODCHEM.2015.05.075.
B. Hu, K. Zhou, Y. Liu, A. Liu, Q. Zhang, G. Han, S. Liu, Y. Yang, Y. Zhu, D. Zhu, Optimization of microwave-assisted extraction of oil from tiger nut (Cyperus esculentus L.) and its quality evaluation, Ind Crops Prod 115 (2018) 290-297. https://doi.org/10.1016/J.INDCROP.2018.02.034.
J.B. Guckert, K.E. Cooksey, L.L. Jackson, Lipid sovent systems are not equivalent for analysis of lipid classes in the microeukaryotic green alga, Chlorella, J Microbiol Methods 8 (1988) 139-149. https://doi.org/10.1016/0167-7012(88)90015-2.
W. Wojnowski, M. Tobiszewski, F. Pena-Pereira, E. Psillakis, AGREEprep - Analytical greenness metric for sample preparation, TrAC Trends in Analytical Chemistry 149 (2022) 116553. https://doi.org/10.1016/J.TRAC.2022.116553.
