Rodríguez-Palmero M, Lopez-Sabater M C, Castellote-Bargallo A I, De la Torre-Boronat M C, Rivero-Urgell M
University of Barcelona, Department of Human Physiological Sciences and Nutrition, Spain.
J Chromatogr A. 1997 Aug 22;778(1-2):435-9. doi: 10.1016/s0021-9673(97)00554-2.
We have validated and compared two methods for the determination of fatty acid profiles in biological samples by capillary gas chromatography. Method I consisted of a previous lipid extraction and esterification of fatty acids using boron trifluoride-methanol. Method II was a direct method that combined extraction and esterification of freeze-dried samples in a single step, using acetyl chloride as the reagent. The two methods were applied to the analysis of plasma and erythrocyte samples. Both methods gave similar results in plasma, whereas in erythrocytes, the direct method gave significantly higher contents of total fatty acids. Precision and recovery rates were determined and the results were satisfactory. Detection and quantification limits showed that both methods had excellent sensitivity. It was concluded that the direct method has substantial advantages over the conventional method, such as higher values in erythrocytes, rapidity and less possibility of contamination or fatty acid losses. Therefore, it is preferable for the analysis of biological samples such as plasma and erythrocytes.
我们已经验证并比较了两种通过毛细管气相色谱法测定生物样品中脂肪酸谱的方法。方法I包括先前的脂质提取以及使用三氟化硼 - 甲醇对脂肪酸进行酯化。方法II是一种直接方法,它使用乙酰氯作为试剂,在一步中对冻干样品进行提取和酯化。这两种方法被应用于血浆和红细胞样品的分析。两种方法在血浆中给出了相似的结果,而在红细胞中,直接法给出的总脂肪酸含量显著更高。测定了精密度和回收率,结果令人满意。检测限和定量限表明两种方法都具有出色的灵敏度。得出的结论是,直接法相对于传统方法具有实质性优势,例如在红细胞中的值更高、速度更快以及污染或脂肪酸损失的可能性更小。因此,它更适合用于血浆和红细胞等生物样品的分析。