[关键词]
[摘要]
目的 建立高效液相色谱测定人和小鼠粪便中短链脂肪酸的方法。方法 样品进行衍生化处理,采用高效液相色谱法测定乙酸、丙酸、丁酸以及戊酸。YMC-Pack FA型色谱柱(250 mm×6.0 mm,5 μm),流动相甲醇-水(60:40,0.1%三氟乙酸调节pH值至4.5),检测波长400 nm,体积流量1.0 mL/min,柱温50℃,进样量20 μL。结果 乙酸、丙酸、丁酸、戊酸分别在0.300~6.005 mg/L(r=0.999 2)、0.370~7.408 mg/L(r=0.998 5)、0.441~8.811 mg/L(r=0.997 3)、0.511~10.210 mg/L(r=0.997 9)线性关系良好;平均加样回收率分别为93.27%、96.40%、95.67%、95.43%,RSD值分别为4.62%、5.42%、3.64%、3.92%。结论 方法操作简单,精密度、稳定性、重复性良好,可用于测定人、小鼠粪便中短链脂肪酸。
[Key word]
[Abstract]
Objective To establish an HPLC method for determination of short chain fatty acids in human and mice feces. Methods All samples were derived, and the contents of acetic acid, propionic acid, butyric acid, and pentanoic acid were determined by HPLC method. YMC-Pack FA chromatographic column (250 mm×6.0 mm, 5 μm) was used. The mobile phase consisted of methanol-water (0.2% trifluoroacetic acid solution regulated pH value to 4.5). The detection wavelengths were set at 400 nm. The flow rate was 1.0 mL/min, temperature of column was set at 50℃, and volume of injection was 20 μL. Results The linear range of acetic acid, propionic acid, butyric acid, and pentanoic acid were 0.300-6.005 mg/L (r=0.999 2), 0.370-7.408 mg/L (r=0.998 5), 0.441-8.811 mg/L (r=0.997 3), and 0.511-10.210 mg/L (r=0.997 9), respectively. The average recoveries were 93.27%, 96.40%, 95.67%, and 95.43%, respectively. The corresponding RSD values were 4.62%, 5.42%, 3.64%, and 3.92%, respectively. Conclusion The method is simple, accurate, stable, and reproducible, which can be used for the determination of short chain fatty acids in human and mice feces.
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[基金项目]
国家自然科学基金资助项目(81470796)