Biomedical research in areas such as metabolic disorders, neuromodulatory, and immunomodulatory conditions involves lipid metabolism and demands a reliable and inexpensive method for quantification of short chain fatty acids (SCFAs). We report a GC–MS method for analysis of all straight-chain and branched-chain SCFAs using pentafluorobenzyl bromide (PFBBr) as derivatization reagent. We optimized the derivatization and GC–MS conditions using a mixture containing all eight SCFA standards, i.e., five straight-chain and three branched-chain SCFAs. The optimal derivatization conditions were derivatization time 90 min, temperature 60 °C, pH 7, and (CH₃)₂CO:H₂O ratio 2:1 (v:v). Comparing the performance of different GC column configurations, a 30 m DB-225ms hyphenated with a 30 m DB-5ms column in tandem showed the best separation of SCFAs. Using the optimized experiment conditions, we simultaneously detected all SCFAs with much improved detection limit, 0.244–0.977 μM. We further applied the developed method to measure the SCFAs in mouse feces and all SCFAs were successfully quantified. The recovery rates of the eight SCFAs ranged from 55.7% to 97.9%.

在诸如代谢紊乱，神经调节和免疫调节条件等领域的生物医学研究涉及脂质代谢，因此需要一种可靠且廉价的方法来定量短链脂肪酸（SCFA）。我们报告了使用五氟苄基溴（PFBBr）作为衍生试剂分析所有直链和支链SCFA的GC-MS方法。我们使用包含所有八种SCFA标准品（即五种直链和三支支链SCFA）的混合物优化了衍生化和GC-MS条件。最佳衍生条件为衍生时间90分钟，温度60°C，pH 7和（CH ₃）₂ CO：H ₂ O比2：1（v：v）。比较不同GC色谱柱配置的性能，将30 m DB-225ms与30 m DB-5ms色谱柱串联起来表现出SCFA的最佳分离。使用优化的实验条件，我们同时检测了所有SCFA，检测限大大提高，为0.244–0.977μM。我们进一步应用了开发的方法来测量小鼠粪便中的SCFA，所有SCFA均已成功定量。八个SCFA的回收率在55.7％至97.9％之间。