Optimization and validation of direct gas chromatography-mass spectrometry method for simultaneous quantification of ten short-chain fatty acids in rat feces

•A direct GC-MS method was developed to quantify ten SCFAs in rat feces.•The detected SCFAs included toxicity-associated formic acid that rarely determined.•Chromatographic/spectrometric conditions and sample preparation approaches were optimized.•The method was validated and applied to quantify SCFAs in fecal samples of animal models.