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~muL of extraction/methylation mixture and 400~muL of heptane, followed by reaction at 80~circC for 2 hours.
시료의 지방산 분석은 먼저 지방의 추출이 선행되어야 하는데 이때에는 시료량 및 시약이 많이 들고 또한 분석시간이 많이 소요되므로 지방 추출 및 지방산의 에스테르화를 한 단계에서 실시하는 One step extraction/methylation법(방법 1)에 따라 주요 유료작물의 지방산 조성을 살펴보았다 이 방법은 methanol-heptane-benzene-DMP-H2 SO4 혼합용매를 사용하여 지방 추출 및 지방산 에스테르화를 단일층에서 일어나게 한것으로 기존의 sodium methoxide를 촉매로 한 methanolysis 법에 의한 지방산 분석 결과와 유의적인 차이를 보이지 않았다. 또한 이 혼합시약으로 이미 추출된 지방을 대상으로 실험하였을 때에도(방법 2) 기존방법과 유의적인 차이가 없었다. 따라서 이 One step extraction/methylation법은 시료가 생체이든 추출된 지방이든 그 과정이 단순하고 시간과 노력이 훨씬 적게 소요되어 지방산 분석에 널리 이용될 수 있을 것이며, 특히 1회에 많은 시료를 처리하고자 할 때 더욱 분석효율이 높을 것으로 기대되었다