title: Simultaneous determination of glutathione, cysteine, homocysteine, and cysteinylglycine in biological fluids by ion-pairing high-performance liquid chromatography coupled with precolumn derivatization.
abstract: Biologically active low-molecular-mass thiols, mainly including glutathione (GSH), cysteine (Cys), homocysteine (Hcy), and cysteinylglycine (Cys-Gly), are important physiological components in biological fluids, and their analytical methods have gained continuous attention over recent years. We developed and validated a novel HPLC method for the quantification of GSH, Cys, Hcy, and Cys-Gly in human plasma, urine, and saliva using 4-chloro-3,5-dinitrobenzotrifluoride as the derivatization reagent. Analyses were linear from 0.15 to 500 μM with the coefficient regression range of 0.9987-0.9994. Detection limits ranged from 0.04 to 0.08 μM (S/N=3). The developed method was applied to quantification of four thiols in human biological fluids collected from five donors with the concentration range of 2.50-124.25 μM, 0-72.81 μM, and 0-4.25 μM for plasma, urine, and saliva, respectively. The present method seemed to be an attractive choice for the determination of thiols in plasma, urine, and saliva.
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University , Beijing, China , 100193.
authors.forename: Wenbing
authors.initials: W
authors.lastname: Zhang
authors.forename: Pingliang
authors.initials: P
authors.lastname: Li
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Geng
authors.forename: Yongheng
authors.initials: Y
authors.lastname: Duan
authors.forename: Mingcheng
authors.initials: M
authors.lastname: Guo
authors.forename: Yongsong
authors.initials: Y
authors.lastname: Cao
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003545:Cysteine / Q000032:analysis* / Q000097:blood / Q000652:urine; D004151:Dipeptides / Q000032:analysis* / Q000097:blood / Q000652:urine; D005978:Glutathione / Q000032:analysis* / Q000097:blood / Q000652:urine; D006710:Homocysteine / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D008297:Male; D012463:Saliva / Q000737:chemistry*; D055815:Young Adult
chemical_list: D004151:Dipeptides; D006710:Homocysteine; C028505:cysteinylglycine; D005978:Glutathione; D003545:Cysteine