Comparative studies on esterification methods for the quantitative determination of fatty acids (FAs) were accomplished using a rotatable central composite design (RCCD). The effect of the three methylation methods on the isomerization of unsaturated fatty acids in gas chromatography analysis was also evaluated. The experimental design showed that the effect of both temperature and duration time of reaction on the recovery of methylation was significant. After optimization, the method was validated and the results for linearity, precision, and limit of detection (LOD) were presented. Methods were similar on account of the fatty acid methyl esters recovery (%) determined (91.30–96.44%). The acid-catalyzed procedures gave no isomerization of conjugated dienes and no artifacts, but artifacts were observed by using a methyl chloroformate reagent. Therefore, the optimized acid-catalyzed methylation process would be more convenient for polyunsaturated fatty acid analysis, whereas methyl chloroformate reagent would be an alternative for fast analysis.

使用可旋转的中心复合设计（RCCD）完成了用于定量测定脂肪酸（FAs）的酯化方法的比较研究。还评估了三种甲基化方法在气相色谱分析中对不饱和脂肪酸异构化的影响。实验设计表明，温度和反应持续时间对甲基化回收率均具有显着影响。优化后，对该方法进行了验证，并给出了线性，精度和检测限（LOD）的结果。由于测定的脂肪酸甲酯的回收率（％）（91.30–96.44％），方法相似。酸催化的方法没有共轭二烯的异构化，也没有伪影，但是通过使用氯甲酸甲酯试剂观察到伪影。