Simultaneous determination of three endogenous chiral thiol compounds in serum from humans at normal and stress states using ultrahigh-performance liquid chromatography coupled to quadrupole-Orbitrap high resolution mass spectrometry

•We have developed a novel chiral mass spectrometry derivatization reagent, NCS-OTPP, for the enantiomeric separation of chiral thiol compounds by UHPLC-Q-Orbitrap HRMS.•A high sensitive, high selective and high accurate method was established for the detection and separation chiral thiol compounds.•Three kinds of free and total chiral thiol compounds in serum of human were detected and D-Hcy was determined in human serum for the first time.•The differences of three kinds of free and total chiral thiol compounds in serum from human at normal and stress states were compared.