柱前衍生化高效液相色谱法测定生物样本中短链脂肪酸及乳酸含量

建立一种适用于同时测定肠内容物、血清等生物样品中短链脂肪酸（short-chain fatty acids，SCFAs）及乳酸含量的方法。使用气相色谱法、高效液相色谱法、柱前衍生化高效液相色谱法检测小鼠粪便中SCFAs及乳酸含量并进行对比分析；对建立的柱前衍生化高效液相色谱法进行方法学评价，并将其应用于肠内容物、血清等生物样品的分析。结果表明：气相色谱法可快速检测乙酸、丙酸、丁酸、戊酸，但不能同时检测乳酸；高效液相色谱法杂峰多，检测乙酸含量误差大，准确度不高；柱前衍生化高效液相色谱法检测范围广，可同时检测肠内容物与血清中乳酸及5 种SCFAs的含量，样品色谱图杂峰少，各组分分离度高，方法准确度高，各组分测定的变异系数均不高于8.87%。在同时检测肠内容物与血清等生物样品中SCFAs及乳酸含量方面，相比于气相色谱法和高效液相色谱法，柱前衍生化高效液相色谱法更具技术优势，更能有效满足当前迅猛发展的微生物组学、代谢组学的复杂样品中对SCFAs及乳酸含量准确分析的应用需求。

Determination of Short-Chain Fatty Acids and Lactic Acid in Biosamples by High Performance Liquid Chromatography with Pre-Column Derivatization

A pre-column derivatization high performance liquid chromatography (PCD-HPLC) method was developed for simultaneous analysis of short-chain fatty acids (SCFAs) and lactic acid in complex biological samples including intestinal contents and serum. The contents of SCFAs and lactate in mouse feces were determined by PCD-HPLC and compared with those determined by gas chromatography (GC) and high performance liquid chromatography (HPLC). Then, methodological evaluation was performed on PCD-HPLC. PCD-HPLC was applied to analyze intestinal contents and blood. Results showed that GC could rapidly detect acetate, propionate, butyrate and valerate, but not lactic acid. HPLC yielded many impurity peaks and inaccurate results for lactic acid. The PCD-HPLC method was applicable to a wide range of samples and allowed for simultaneous analysis of lactate and five SCFAs in intestinal contents and serum. Only a few impurity peaks appeared in the chromatogram, and a good separation of all analytes was achieved. PCD-HPLC was accurate, giving a coefficient of variation (CV) less than or equal to 8.87% for each analyte. In conclusion, this method is superior to GC and HPLC in determining SCFAs and lactic acid in complicated biological samples such as intestinal contents and serum, and can meet the demand for accurate analysis of SCFAs and lactic acid in complicated biological samples in response to the rapid development of microbial metabolomics.