Determination of short-chain fatty acids in human serum by gas chromatography-mass spectrometry with pre-column benzyl esterification
OBJECTIVE To establish a method for determining short-chain fatty acids(SCFAs)in human serum samples by gas chromatography-mass spectrometry(GC-MS).METHODS BnOH-Py solvent mixture 200 μL(3:2,V/V)and dimethyl sulfoxide(DMSO)100 μL were subsequently added int0 200 μL sample(20 μL IS)and vortexed for 30 s.Then 100 μL benzyl chloroformate(BCF)was slowly added into reaction system and vortexed for 3 min at room temperature,followed by extracted with 200 μL cyclohexane.Then 100 μL derivative extract(upper cyclohexane layer)was transferred into a glass insert and injected into a GC-MS instrument for analysis.RESULTS Twelve SCFAs were separated distinctly and five SCFAs were detected in human sera including formic acid,acetic acid,propionic acid,iso-butyric acid and butyric acid.Derivation with BCF at room temperature showed an excellent linearity(R2＞0.992 5)and excellent recoveries(89.00％-114.83％)under low/moder ate/high concentration.Intra/inter-day precision was achieved within a range of 0.90％to 13.66％(n=5).Low limit of detec-tions(LODs)of different SCFAs ranged from 0.02 to 0.30 μg·mL-1.Successful application of this method on clinical serum samples was demonstrated in 30 volunteers.CONCLUSION A simple,fast and sensitive method has been developed for measur-ing SCFAs in human sera.
short-chain fatty acidspre-column derivationgas chromatography coupled with mass spectrometry detector(GC-MS)serum
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