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Fast Transmethylation of Total Lipids in Dried Blood by Microwave Irradiation and its Application to a Population Study
Author(s) -
Lin Yu Hong,
Hanson Jennifer A.,
Strandjord Sarah E.,
Salem Nicholas M.,
Dretsch Michael N.,
Haub Mark D.,
Hibbeln Joseph R.
Publication year - 2014
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-014-3918-3
Subject(s) - transmethylation , polyunsaturated fatty acid , butylated hydroxytoluene , docosahexaenoic acid , chemistry , linoleic acid , population , fatty acid , lipidology , chromatography , arachidonic acid , biochemistry , antioxidant , food science , medicine , environmental health , amino acid , methionine , enzyme
A methodology combining finger‐pricked blood sampling, microwave accelerated fatty acid assay, fast gas chromatography data acquisition, and automated data processing was developed, evaluated and applied to a population study. Finger‐pricked blood was collected on filter paper previously impregnated with 0.05 mg of the antioxidant butylated hydroxytoluene and air‐dried at room temperature. Transmethylation was accelerated by microwave irradiation in an explosion‐proof multimode microwave reaction system. The chemical procedure was based on a one‐step direct transmethylation procedure catalyzed by acetyl chloride. The short‐term stability of PUFA in blood dried on filter paper and storage at room temperature was examined using venous blood. The recoveries ranged from 97 to 101 % for the categorized fatty acids as well as the ratios of n‐6 to n‐3 PUFA and the n‐3 % highly unsaturated fatty acid. Specifically, recoveries were 99, 98, 97, and 97 % for linoleic acid (18:2n‐6), arachidonic acid (ARA), α‐linolenic acid (ALA), and docosahexaenoic acid (DHA), respectively. The mol% (mean ± SD, 95 % confidence interval) of fatty acid composition in subjects from the population study was determined as 36.2 ± 3.8 (35.8, 36.7), 23.2 ± 3.0 (22.8, 23.5), 36.8 ± 3.5 (36.4, 37.2) and 3.79 ± 1.0 (3.68, 3.91) for the saturated, monounsaturated, n‐6 and n‐3 PUFA, respectively. Individually, the mean mol% (95 % CI) was 22.6 (22.3, 22.9) for 18:2n‐6, 9.5 (9.3, 9.7) for ARA, 0.51 (0.49, 0.53) for ALA, 0.42 (0.38, 0.47) for eicosapentaenoic acid (EPA), and 1.67 (1.61, 1.73) for DHA. This methodology provides an accelerated yet high‐efficiency, chemically safe, and temperature‐controlled transmethylation, with diverse laboratory applications including population studies.