建立了气相色谱法(GC)同时检测人唾液中7种短链脂肪酸(Short-chain fatty acids, SCFAs)含量的方法。唾液样品与乙醇溶液按体积比1:1混合(含0.5%(V/V)浓HCl)涡旋混合,离心后取上清液进行GC分析。采用DB-FFAP毛细管柱(30 m×0.25 mm×0.25μm)进行分离;氢火焰离子化检测器(FID)进行检测,内标法定量。实验结果表明,唾液中共检测到7种SCFAs(乙酸、丙酸、异丁酸、丁酸、异戊酸、戊酸、己酸),7种SCFAs与内标物2-乙基丁酸的色谱峰峰面积比值与其浓度均呈现良好的线性关系(R2≥0.999),检出限(LOD,S/N=3)和定量限(LOQ,S/N=10)分别为0.060~0.198μg/mL和0.180~0.594μg/mL,平均加样回收率为94.8%~109.7%(RSD≤4.3%)。本方法简单、快速、准确,可用于测定人的唾液中短链脂肪酸的含量。
A gas chromatographic (GC) method was established for the simultaneous analysis of seven short-chain fatty acids (SCFAs) including acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid and hexanoic acid in human saliva. Salivary sample was diluted by ethanol solution by ratio of 1:1 (V/V), with 0.5% (V/V) hydrochloric acid in sample solution, then vortexed and centrifuged. The supernatent was subjected to GC analysis. The chromatographic separation was performed on a DB-FFAP capillary column (30 m×0.25 mm×0.25 μm) using flame ionization detector (FID). The quantification was achieved by internal standard method. The experimental results showed that an excellent correlation coefficient (R2≥0.999) was obtained for all the calibration curves of seven SCFAs. The limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) were 0.060-0.198 μg/mL and 0.180-0.594 μg/mL, respectively. The average recoveries were 94.8%-109.7% with relative standard deviation (RSD)≤4.3% (n=6). The developed method is simple, rapid and accurate, and suitable for the simultaneous determination of seven SCFAs in human saliva.
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