as an IS and CH3CN (10L). Then, 30L perchloric acid (60%) was added for deproteinization, and 800L chloroform, 100L nheptane, and 100L H2O were added for the extraction of fatty acids. After the solution was centrifuged at 3000×g for 5 min, the organic layer (400L) was transferred to a tube and evaporated under reduced pressure without heating for 22 min. Following the addition of 10mMp-toluenesulfonic acid (PTSA) in MeOH (200L) to the dried residue and incubation at 62 ◦C for 2 h for methyl esterification, H2O (200L) and n-hexane (200L) were sequentially added, mixed for 2min, and centrifuged at 4000×g for 5min. The hexane layer (150L), which included fatty acid methyl esters (FAMEs), was obtained. Then, 10L sample solution was subjected
to GC–MS analysis.
as an IS and CH3CN (10L). Then, 30L perchloric acid (60%) was added for deproteinization, and 800L chloroform, 100L nheptane, and 100L H2O were added for the extraction of fatty acids. After the solution was centrifuged at 3000×g for 5 min, the organic layer (400L) was transferred to a tube and evaporated under reduced pressure without heating for 22 min. Following the addition of 10mMp-toluenesulfonic acid (PTSA) in MeOH (200L) to the dried residue and incubation at 62 ◦C for 2 h for methyl esterification, H2O (200L) and n-hexane (200L) were sequentially added, mixed for 2min, and centrifuged at 4000×g for 5min. The hexane layer (150L), which included fatty acid methyl esters (FAMEs), was obtained. Then, 10L sample solution was subjectedto GC–MS analysis.
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