Literature Survey

Combined Extraction-Esterification of Fatty Acids

Everyone likes a shortcut, so successful extraction of lipids with simultaneous transesterification is an attractive idea. Many methods have been described in the literature. However, I would like to add one caveat. Most extraction methods have a washing step that gets rid of many potential contaminants; eliminating this step can give rise to artefact peaks on gas chromatography traces. You may find my review article on methods of transesterification of value.

  • Christie,W.W. Preparation of ester derivatives of fatty acids for chromatographic analysis. In Advances in Lipid Methodology - Two, pp. 69-111 (1993) (edited by W.W. Christie, Oily Press, Dundee).

This is now available on our website here.

In addition, there is a useful review of the methodology by Carrapiso et al. cited below. The following references, covering the years 1990 to the present, were collected as part of our regular literature reviews for our own research purposes. We cannot claim to have covered the subject exhaustively, but we have done our best. References are listed alphabetically by the first author of a paper, but not necessarily chronologically by that author.

  • Abdulkadir,S. and Tsuchiya,M. One-step method for quantitative and qualitative analysis of fatty acids in marine animal samples. J. Exp. Marine Biol. Ecol., 354, 1-8 (2008).
  • Akoto,L., Pel,R., Irth,H., Brinkman,U.A.T. and Vreuls,R.J.J. Automated GC-MS analysis of raw biological samples - Application to fatty acid profiling of aquatic micro-organisms. J. Anal. Appl. Pyrol., 73, 69-75 (2005).
  • Akoto,L., Stellaard,F., Irth,H., Vreuls,R.J. and Pel,R. Improved fatty acid detection in micro-algae and aquatic meiofauna species using a direct thermal desorption interface combined with comprehensive gas chromatography-time-of-flight mass spectrometry. J. Chromatography A, 1186, 254-261 (2008).
  • Akoto,L., Vreuls,RJJ., Irth,H., Pel,R. and Stellaard,F. Fatty acid profiling of raw human plasma and whole blood using direct thermal desorption combined with gas chromatography-mass spectrometry. J. Chromatography A, 1186, 365-371 (2008).
  • Ali,L.H., Angyal,G., Weaver,C.M. and Rader,J.I. Comparison of capillary column gas chromatographic and AOAC gravimetric procedures for total fat and distribution of fatty acids in foods. Food Chem., 58, 149-160 (1997).
  • Alves,S.P., Cabrita,A.R.J., Fonseca,A.J.M. and Bessa,R.J.B. Improved method for fatty acid analysis in herbage based on direct transesterification followed by solid-phase extraction. J. Chromatogr. A, 1209, 212-219 (2008).
  • Armstrong,J.M., Metherel,A.H. and Stark,K.D. Direct microwave transesterification of fingertip prick blood samples for fatty acid determinations. Lipids, 43, 187-196 (2008).
  • Ashraf-Khorassani,M., Ude,M., Doane-Weideman,T., Tomczak,J. and Taylor,L.T. Comparison of gravimetry and hydrolysis/derivatization/gas chromatography-mass spectrometry for quantitative analysis of fat from standard reference infant formula powder using supercritical fluid extraction. J. Agric. Food Chem., 50, 1822-1826 (2002).
  • Bailey-Hall,E., Nelson,E.B. and Ryan,A.S. Validation of a rapid measure of blood PUFA levels in humans. Lipids, 43, 181-186 (2008).
  • Basile,F.., Beverly,M.B., Abbas-Hawks,C., Mowry,C.D., Voorhees,K.J. and Hadfield,T.L. Direct mass spectrometric analysis of in situ thermally hydrolyzed and methylated lipids from whole bacterial cells. Anal. Chem., 70, 1555-1562 (1998).
  • Bell,J.G., Mackinlay,E.E., Dick,J.R., Younger,I., Lands,B. and Gilhooly,T. Using a fingertip whole blood sample for rapid fatty acid measurement: method validation and correlation with erythrocyte polar lipid compositions in UK subjects. British J. Nutr., 106, 1408-1415 (2011)
  • Bigelow,N.W., Hardin,W.R., Barker,J.P., Ryken,S.A., MacRae,A.C. and Cattolico,R.A. A comprehensive GC-MS sub-microscale assay for fatty acids and its applications. J. Am. Oil Chem. Soc., 88, 1329-1338 (2011).
  • Bohnert,B., Braun,M., Winter,H. and Fluck,B. Direct esterification method for analysis of long-chain polyunsaturated fatty acids (LC-PUFAs) in infant formulae. Res. Techn., 204, 27-31 (1997).
  • Brumback,T.B., Hazebroek,J., Lamb,D., Danielson,L. and Orman,B. Automated fatty acid analysis from seeds - from field samples to data-bases. Chemom. Intell. Lab. Systems, 21, 215-222 (1993).
  • Buhlmann,R., Carmona,J., Donzel,A., Donzel,N., Gil,J., Haab,J.M. and Hamilton,S.D. A robotic system for the extraction and esterification of bacterial fatty acids. Am. Lab., 25 (No. 9), 18-19 (1993).
  • Buyer,J.S. Rapid and sensitive FAME analysis of bacteria by cold trap injection gas chromatography. J. Microbiol. Methods, 67, 187-190 (2006).
  • Cantellops,D., Reid,A.P., Eitenmiller,R.R. and Long,A.R. Determination of lipids in infant formula powder by direct extraction methylation of lipids and fatty acid methyl esters (FAME) analysis by gas chromatography. J. Ass. Off. Anal. Chem. Int., 82, 1128-1139 (1999).
  • Carrapiso, A..I, Timon,M.L, Petron,M.J., Tejeda,J.F. and Garcia,C. In situ transesterification of fatty acids from Iberian pig subcutaneous adipose tissue. Meat Sci., 56, 159-164 (2000).
  • Carrapiso, A.I. and Garcia,C. Development in lipid analysis: Some new extraction techniques and in situ transesterification. Lipids, 35, 1167-1177 (2000).
  • Christie, W.W. and Han, X. Lipid Analysis - Isolation, Separation, Identification and Lipidomic Analysis (4th edition), 446 pages (Oily Press, Bridgwater, U.K. and Woodhead Publishing Ltd, Cambridge, U.K.) (2010) - Woodhead Publishing Ltd.
  • Christophe, A. Fatty acid profiling in lipid disorders. In: Advances in Lipid Methodology - Five, pp. 255-272 (ed. R.O. Adlof, Oily Press, Bridgwater) (2003).
  • de La Cruz Garcia, C., Hernandez,J.L. and Lozano,J.S. Gas chromatographic determination of the fatty-acid content of heat-treated green beans. J. Chromatogr. A, 891, 367-370 (2000).
  • Dickey, L.A., Teter,B.B., Sampugna,J. and Woods,L.C. Comparison of a direct transesterification method and the Bligh and Dyer method to determine fatty acid content in striped bass tissues and diet. North Am. J. Aquaculture, 64, 158-163 (2002).
  • Dionisi,F., Golay,P.A., Elli,M. and Fay,L.B. Stability of cyclopropane and conjugated linoleic acids during fatty acid quantification in lactic acid bacteria. Lipids, 34, 1107-1115 (1999).
  • Dworzanski,J.P., Berwlad,L. and Meuzelaar,H.L.C. Pyrolytic methylation-gas chromatography of whole bacterial cells for rapid profiling of cellular fatty acids. Appl. Environm. Microbiol., 56, 1717-1724 (1990).
  • Garces,R. and Mancha,M. One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal. Biochem., 211, 139-143 (1993).
  • Garcia-Barcelo,M., Luquin,M., Belda,F. and Ausina,V. GC whole-cell fatty acid analysis as an aid for the identification of mixed Mycobacterial cultures. J. Chromatogr. B, 617, 299-303 (1993).
  • Gharaibeh,A.A. and Voorhees,K.J. Characterization of lipid fatty acids in whole-cell microorganisms using in situ supercritical-fluid derivatization/extraction and gas chromatography-mass spectrometry. Anal. Chem., 68, 2805-2810 (1996).
  • Glaser,C., Demmelmair,H. and Koletzko,B. High-throughput analysis of fatty acid composition of plasma glycerophospholipids. J. Lipid Res., 51, 216-221 (2010).
  • Glaser,C., Demmelmair,H. and Koletzko,B. High-throughput analysis of total plasma fatty acid composition with direct in situ transesterification. PLOS One, 5, e12045 (2010).
  • Golay,P.A., Dionisi,F., Hug,B., Giuffrida,F. and Destaillats,F. Direct quantification of fatty acids in dairy powders with special emphasis on trans fatty acid content. Food Chem., 101, 1115-1120 (2007).
  • Gomez-Brandon,M., Lores,M. and Dominguez,J. Comparison of extraction and derivatization methods for fatty acid analysis in solid environmental matrixes. Anal. Bioanal. Chem., 392, 505-514 (2008).
  • Griffiths,M.J., van Hille,R.P. and Harrison,S.T.L. Selection of direct transesterification as the preferred method for assay of fatty acid content of microalgae. Lipids, 45, 1053-1060 (2010).
  • Guillou,A., Soucy,P. and Khalil,M. Preparation of fatty acid methyl esters from brook charr tissues - comparison of a classical and a direct method. Fett-Lipid, 98, 18-21 (1996).
  • Husek,P., Simek,P. and Tvrzická,E. Simple and rapid procedure for the determination of individual free fatty acids in serum. Anal. Chim. Acta, 465, 433-439 (2002).
  • Juarez,M., Polvillo,O., Conto,M., Ficco,A., Ballico,S. and Failla,S. Comparison of four extraction/methylation analytical methods to measure fatty acid composition by gas chromatography in meat. J. Chromatogr. A, 1190, 327-332 (2008).
  • Kaushik,N. and Agnihotri,A. Evaluation of improved method for determination of rapeseed-mustard FAMEs by GC. Chromatographia, 44, 97-99 (1997).
  • Khoomrung,S., Chumnanpuen, P., Jansa-ard, S., Nookaew, I. and Nielsen, J. Fast and accurate preparation fatty acid methyl esters by microwave-assisted derivatization in the yeast Saccharomyces cerevisiae. Appl. Microbiol. Biotechnol., 94, 1637-1646 (2012).
  • Klingler,M. and Koletzko, B. Novel methodologies for assessing omega-3 fatty acid status - a systematic review. Brit. J. Nutr., 107, S53-S63 (2012).
  • Koirala,B. and Rosentreter,J. Examination of prehistoric artifacts via fatty acid methyl ester (FAME) techniques using modern environmental stewardship. J. Archaeol. Sci., 36, 1229-1242 (2009).
  • Li,J.J., Yue,Y.X., Hu,X.J. and Zhong,H.Y. Rapid transmethylation and stable isotope labeling for comparative analysis of fatty acids by mass spectrometry. Anal. Chem., 81, 5080-5087 (2009).
  • Li,J.J., Yue,Y.X., Li,T.T., Hu,X.J. and Zhong,H.Y. Gas chromatography-mass spectrometric analysis of bonded long chain fatty acids in a single zebrafish egg by ultrasound-assisted one-step transmethylation and extraction. Anal. Chim. Acta, 650, 221-226 (2009).
  • Li,Y., Beisson,F., Pollard,M. and Ohlrogge,J. Oil content of Arabidopsis seeds: The influence of seed anatomy, light and plant-to-plant variation. Phytochemistry, 67, 904-915 (2006).
  • Liu,Y.Q., Cho,S.R. and Danielson,N.D. Solid-phase microextraction and on-line methylation gas chromatography for aliphatic carboxylic acids. Anal. Bioanal. Chem., 373, 64-69 (2002).
  • Lu,Y. and Harrington,P.B. Classification of bacteria by simultaneous methylation-solid phase microextraction and gas chromatography/mass spectrometry analysis of fatty acid methyl esters. Anal. Bioanal. Chem., 397, 2959-2966 (2010).
  • Marangoni,F., Colombo,C. and Galli,C. A method for the direct evaluation of the fatty acid status in a drop of blood from a fingertip in humans: applicability to nutritional and epidemiological studies. Anal. Biochem., 326, 267-272 (2004).
  • Marx,F. and Stender,A. Rapid micro methods for the GLC determination of fat content and fatty acid composition in seeds rich in lipids. Fett-Lipid, 99, 25-28 (1997).
  • Mayberry,W.R. and Lane,J.R. Sequential alkaline saponification/acid hydrolysis/esterification: a one-tube method with enhanced recovery of both cyclopropane and hydroxylated fatty acids. J. Microb. Methods, 18, 21-32 (1993).
  • Mazalli,M.R. and Bragagnolo,N. Validation of two methods for fatty acids analysis in eggs. Lipids, 42, 483-490 (2007).
  • McNichol,J., MacDougall, K.M., Melanson, J.E. and McGinn, P.J. Suitability of soxhlet extraction to quantify microalgal fatty acids as determined by comparison with in situ transesterification. Lipids, 47, 195-207 (2012).
  • Meier,S., Mjos,S.A., Joensen,H. and Grahl-Nielsen,O. Validation of a one-step extraction/methylation method for determination of fatty acids and cholesterol in marine tissues. J. Chromatogr. A, 1104, 291-298 (2006).
  • Mjos,S.A., Meier,S. and Grahl-Nielsen,O. Geometrical isomerisation of double bonds in acid-catalysed preparation of fatty acid methyl esters. Eur. J. Lipid Sci. Technol., 108, 315-322 (2006).
  • Muller,K.-D., Husmann,H., Nalik,H.P. and Schomburg,G. Transesterification of fatty acids from microorganisms and human blood serum by trimethylsulfonium hydroxide (TMSH) for GC analysis. Chromatographia, 30, 245-248 (1990).
  • Muller,K.D., Nalik,H.P., Schmid,E.N., Husmann,H. and Schomburg,G. Fast identification of Mycobacterium species by GC analysis with trimethylsulfonium hydroxide (TMSH) for transesterification. J. High Resolut. Chromatogr., 16, 161-165 (1993).
  • O'Fallon,J.V., Busboom,J.R., Nelson,M.L. and Gaskins,C.T. A direct method for fatty acid methyl ester synthesis: Application to wet meat tissues, oils, and feedstuffs. J. Animal Sci., 85, 1511-1521 (2007).
  • Palmquist,D.L. and Jenkins,T.C. Challenges with fats and fatty acid methods. J. Anim. Sci., 81, 3250-3254 (2003).
  • Pan,L. and Pawliszyn,J. Derivatization/solid-phase microextraction: New approach to polar analytes. Anal. Chem., 69, 196-205 (1997).
  • Park,P.W. and Goins,R.E. In situ preparation of fatty acid methyl esters for analysis of fatty acid composition in foods. J. Food Sci., 59, 1262-1266 (1994).
  • Paterson,E. and Amado,R. Simplified method for the simultaneous gas chromatographic determination of fatty acid composition and cholesterol in food. Food Sci. Technol., 30, 202-209 (1997).
  • Poerschmann,J. and Carlson,R. New fractionation scheme for lipid classes based on "in-cell fractionation" using sequential pressurized liquid extraction. J. Chromatogr. A, 1127, 18-25 (2006).
  • Poerschmann,J., Trommler,U., Biedermann,W., Truyen,U. and Lucker,E. Sequential pressurized liquid extraction to determine brain-originating fatty acids in meat products as encephalopathy risk markers in bovine spongiform assessment studies. J. Chromatogr. A, 1127, 26-33 (2006).
  • Purcaro,G., Tranchida,P.Q., Dugo,P., La Camera,E., Bisignano,G., Conte,L. and Mondello,L. Characterization of bacterial lipid profiles by using rapid sample preparation and fast comprehensive two-dimensional gas chromatography in combination with mass spectrometry. J. Sep. Sci., 33, 2334-2340 (2010).
  • Rader,J.I., Angyal,G., O'Dell,R.G., Weaver,C.M., Sheppard,A.J. and Bueno,M.P. Determination of total fat and saturated fat in foods by packed-column gas-liquid chromatography after acid hydrolysis. Food Chem., 54, 419-427 (1995).
  • Risé,P., Salvetti,F. and Galli,C. Application of a direct transmethylation method to the analysis of fatty acid profile in circulating and cultured cells. Anal. Biochem., 346, 182-184 (2005).
  • Rizzo,A.M., Montorfano,G., Negroni,M., Adorni,L., Berselli,P., Corsetto,P., Wahle,K. and Berra,B. A rapid method for determining arachidonic: eicosapentaenoic acid ratios in whole blood lipids: correlation with erythrocyte membrane ratios and validation in a large Italian population of various ages and pathologies. Lipids in Health and Disease, 9, 7 (2010).
  • Robinson,J.E., Singh,R. and Kays,S.E. Evaluation of an automated hydrolysis and extraction method for quantification of total fat, lipid classes and trans fat in cereal products. Food Chem., 107, 1144-1150 (2008).
  • Rodriguez-Palmero,M., Lopez-Sabater,M.C., Castellote-Bargallo,A.I., Dela Torre-Boronat,M.C. and Rivero-Urgell,M. Comparison of two methods for the determination of fatty acid profiles in plasma and erythrocytes. J. Chromatogr. A, 778, 435-439 (1997).
  • Ruiz-Jimenez,J., Priego-Capote,F. and de Castro,M.D.L. Identification and quantification of trans fatty acids in bakery products by gas chromatography-mass spectrometry after dynamic ultrasound-assisted extraction. J. Chromatogr. A, 1045, 203-210 (2004).
  • Ruiz-Lopez,N., Martinez-Force,E. and Garces,R. Sequential one-step extraction and analysis of triacylglycerols and fatty acids in plant tissues. Anal. Biochem., 317, 247-254 (2003).
  • Rule,D.C. Direct transesterification of total fatty acids of adipose tissue, and of freeze-dried muscle and liver with boron-trifluoride in methanol. Meat Sci., 46, 23-32 (1997).
  • Sanchez-Avila,N., Mata-Granados,J.M., Ruiz-Jimenez,J. and Luque de Castro,M.D. Fast, sensitive and highly discriminant gas chromatography-mass spectrometry method for profiling analysis of fatty acids in serum. J. Chromatogr. A, 1216, 6864-6872 (2009).
  • Sattler,W., Puhl,W., Hayn,M., Kostner,G.M. and Esterbauer,H., Determination of fatty acids in the main lipoprotein classes by capillary GC:BF3/methanol transesterification of lyophilized samples instead of Folch extraction gives higher yields. Anal. Biochem., 198, 184-190 (1991).
  • Schreiner,M. Optimization of solvent extraction and direct transmethylation methods for the analysis of egg yolk lipids. Int. J. Food Prop., 9, 573-581 (2006).
  • Shirai,N., Suzuki,H. and Wada,S. Direct methylation from mouse plasma and from liver and brain homogenates. Anal. Biochem., 343, 48-53 (2005).
  • Snyder,J.M., King,J.W. and Jackson,M.A. Analytical supercritical fluid extraction with lipase catalysis: Conversion of different lipids to methyl esters and effect of moisture. J. Am. Oil Chem. Soc., 74, 585-588 (1997).
  • Sonnichsen,M. and Muller,B.W. A rapid and quantitative method for total fatty acid analysis of fungi and other biological samples. Lipids, 34, 1347-1349 (1999).
  • Suter,B., Grob,K. and Pacciarelli,B. Determination of fat content and fatty acid composition through 1-min. transesterification in the food sample: principles. Z. Lebensm.-Unters. Forsch. A., 204, 252-258 (1997).
  • Takeuchi,T., Ackman,R.G. and Lall,S.P. Differences in fatty acid composition of fish faeces as determined by two extraction methods. J. Sci. Food Agric., 56, 259-264 (1991).
  • Thiemann,G.W., Budge,S.M. and Iverson,S.J. Determining blubber fatty acid composition: A comparison of in situ direct and traditional methods. Marine Mammal Sci., 20, 284-295 (2004).
  • Tomas,A., Tor,M., Villorbina,G., Canela,R., Balcells,M. and Eras,J. A rapid and reliable direct method for quantifying meat acylglycerides with monomode microwave irradiation. J. Chromatogr. A, 1216, 3290-3295 (2009).
  • Ulberth,F. and Henninger,M. Determination of the fatty acid profile of fish by a one-step extraction methylation method. Fat Sci. Technol., 97, 77-80 (1995).
  • Ulberth,F. and Henninger,M. One-step extraction/methylation method for determining the fatty acid composition of processed foods. J. Am. Oil Chem. Soc., 69, 174-177 (1992).
  • Weston,T.R., Derner,J.D., Murrieta,C.M., Rule,D.C. and Hess,B.W. Comparison of catalysts for direct transesterification of fatty acids in freeze-dried forage samples. Crop Sci., 48, 1636-1641 (2008).
  • Xiao, L.P., Mjos, S.A. and Haugsgjerd, B.O. Efficiencies of three common lipid extraction methods evaluated by calculating mass balances of the fatty acids. J. Food Comp. Anal., 25, 198-207 (2012).
  • Xu,M., Basile,F., and Voorhees K.J. Differentiation and classification of user-specified bacterial groups by in situ thermal hydrolysis and methylation of whole bacterial cells with tert-butyl bromide chemical ionization ion trap mass spectrometry. Anal. Chim. Acta, 418, 119-128 (2000).

 

Updated: February 27, 2013