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Fast and accurate preparation fatty acid methyl esters by microwave-assisted derivatization in the yeast Saccharomyces cerevisiae

Sakda Khoomrung (Institutionen för kemi- och bioteknik, Systembiologi) ; Pramote Chumnanpuen (Institutionen för kemi- och bioteknik, Systembiologi) ; Suwanee Jansa-Ard (Institutionen för kemi- och bioteknik, Systembiologi) ; Intawat Nookaew (Institutionen för kemi- och bioteknik, Systembiologi) ; Jens B. Nielsen (Institutionen för kemi- och bioteknik, Systembiologi)
Applied Microbiology and Biotechnology (0175-7598). Vol. 94 (2012), 6, p. 1637-1646.
[Artikel, refereegranskad vetenskaplig]

We present a fast and accurate method for preparation of fatty acid methyl esters (FAMEs) using microwave-assisted derivatization of fatty acids present in yeast samples. The esterification of free/bound fatty acids to FAMEs was completed within 5 min, which is 24 times faster than with conventional heating methods. The developed method was validated in two ways: (1) through comparison with a conventional method (hot plate) and (2) through validation with the standard reference material (SRM) 3275-2 omega-3 and omega-6 fatty acids in fish oil (from the Nation Institute of Standards and Technology, USA). There were no significant differences (P > 0.05) in yields of FAMEs with both validations. By performing a simple modification of closed-vessel microwave heating, it was possible to carry out the esterification in Pyrex glass tubes kept inside the closed vessel. Hereby, we are able to increase the number of sample preparations to several hundred samples per day as the time for preparation of reused vessels was eliminated. Pretreated cell disruption steps are not required, since the direct FAME preparation provides equally quantitative results. The new microwave-assisted derivatization method facilitates the preparation of FAMEs directly from yeast cells, but the method is likely to also be applicable for other biological samples.

Nyckelord: Fatty acid methyl esters, Fatty acid analysis, Microwave-assisted derivatization, Saccharomyces, chromatography/mass spectrometry analysis, direct transesterification, samples, extraction, lipids, gene, cell

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Denna post skapades 2012-07-02. Senast ändrad 2014-10-27.
CPL Pubid: 159872


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Institutionen för kemi- och bioteknik, Systembiologi (2008-2014)


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