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High Throughput Fatty Acid Analysis
Author(s) -
Masood Athar M,
Salem Norman
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a265-c
Subject(s) - transesterification , reagent , chromatography , fatty acid , fatty acid methyl ester , sample preparation , chemistry , throughput , extraction (chemistry) , hexane , centrifugation , neutralization , biodiesel , organic chemistry , computer science , catalysis , telecommunications , antibody , immunology , wireless , biology
Essential fatty acids are the subject of many clinical trials as they are used in disease treatment and also in cross‐sectional analysis since as biomarkers. This work describes both the development of fast GC method where sample chromatographic run time was reduced to 15 min/sample, and simpler fatty acid methyl ester transesterification methods as applicable both to the wet laboratory and to a new high throughput robotic‐based automated method. Accurate quantitative results were obtained by GC analysis of plasma peak concentrations using an internal standard in comparison to traditional (Lepage & Roy) methodology. The robotic method simplification involves the elimination of the multi‐level addition of different reagents, neutralization, vortexing, centrifugation and hexane phase concentration steps. Samples are heated in open tubes for 2.5 hrs at 80°C with addition of toluene and multiple additions of the reagent mixture. The fatty acids were then extracted by two phase extraction via repeated aspiration‐dispensing. These developments, together with data analysis software, make possible for the first time a high throughput system for fatty acid analyses.