Optimization of One-step Extraction/Methylation Method for Analysis of Fatty Acid Composition in Brown Rice

Traditionally fatty acid composition used to be analysed by a GC and the sample preparation process includes lipid extraction from sample and subsequent methyl esters preparation, which are time-consuming and cumbersome. As an alternative, simultaneous extraction/methylation methods are being developed for rapid and simplified sample preparation. To optimize one-step extraction/methylation method for analysis of fatty acid composition in brown rice, various reaction factors such as sample to reaction solution ratio, reaction time and temperature, shaking intensity were changed and resultant fatty acid composition data were evaluated in comparison with previous reports. The ratio of sample weight to reaction solution volume was the most critical factor in that higher sample to reaction solution ratio caused overestimation of palmitic acid and linoleic acid composition, resulting in underestimation of oleic acid. Lower reaction temperature also induced overestimation of linoleic acid and underestimation of oleic acid. Reaction duration and the intensity of shaking prior to and during the reaction, however, caused no significant changes in analysis results. In conclusion, the optimum condition was mixing 5 grains (about 0.2 g) of brown rice with $680{\mu}L$ of extraction/methylation mixture and $400{\mu}L$ of heptane, followed by reaction at $80^{\circ}C$ for 2 hours.