Comparison of different methylation methods for the analysis of conjugated linoleic acid isomers by silver ion HPLC in beef lipids.

Four different methods for the methylation of conjugated linoleic acid isomers (CLA) in ruminant lipids were compared by silver ion (Ag+) HPLC. The combination of base-catalyzed methods followed by an acid-catalyzed method with BF3/MeOH was tested under different temperatures (room temperature and at 60 degrees C), along with based-catalyzed methylation with NaOCH3 and methylation with BF3/MeOH after saponification with NaOH. The comparison among these four methods was done on muscle and adipose tissue samples from bulls. The repeatability theta of the combined base- and acid-catalyzed methylation (NaOCH3/BF3) at ambient temperature for 20 min and at 60 degrees C for 10 min was most suitable for the quantitative Ag+-HPLC analysis of CLA isomers. At 60 degrees C the combined methods supplied the highest concentrations of most CLA isomers. The base-catalyzed methylation and the saponification followed by BF3/MeOH methylation for 5 min generated significantly lower concentrations for most CLA isomers compared to the combined methods.