Simultaneous quantification of straight-chain and branched-chain short chain fatty acids by gas chromatography mass spectrometry

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 br...

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Bibliographic Details
Published in:	Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-08, Vol.1092, p.359-367
Main Authors:	He, Liqing, Prodhan, Md Aminul Islam, Yuan, Fang, Yin, Xinmin, Lorkiewicz, Pawel K., Wei, Xiaoli, Feng, Wenke, McClain, Craig, Zhang, Xiang
Format:	Article
Language:	eng
Subjects:	Animals Fatty Acids, Volatile - analysis Fatty Acids, Volatile - chemistry Fatty Acids, Volatile - metabolism Feces - chemistry Gas Chromatography-Mass Spectrometry - methods GC–MS Hyphenated GC column Limit of Detection Linear Models Male Metabolomics Metabolomics - methods Mice Mice, Inbred C57BL PFBBr Reproducibility of Results Short chain fatty acids
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Summary:	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%. •A method capable of simultaneous quantification of all SCFAs by GC–MS•A DB-225ms and a DB-5ms columns were connected in tandem for separation.•None of the solvent peaks overlaps with the SCFA peaks.•The best LOD and LOQ were achieved for detecting all SCFAs from biosamples.
ISSN:	1570-0232 1873-376X