Pre-column derivatization high-performance liquid chromatographic method for determination of cysteine, cysteinyl-glycine, homocysteine and glutathione in plasma and cell extracts.
A sensitive high-performance liquid chromatographic method for quantification of sulphydryl and disulfide amino acids in human plasma using ultra violet spectrophotometric detection was developed. Precolumn derivatization with 5,5'-dithio-bis-nitrobenzoic acid (DTNB) and an optional pre-derivatization reaction with dithiothreitol allowed both quantitative reduction of disulfides for measurement of total amino acid levels and the measurement of the reduced forms. A dynamic range of 500 nmol/l-750 micromol/l allowed the major analytes of interest to be quantified in plasma without sample dilution. The assay is a sensitive and precise method for the determination of sulphydryl and disulfide amino acids in plasma and cell extracts.
Katrusiak
A E
AE
Department of Pathology, College of Medicine, University of Saskatchewan, Saskatoon, Canada.
Paterson
P G
PG
Kamencic
H
H
Shoker
A
A
Lyon
A W
AW
D002457:Cell Extracts / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000032:analysis* / Q000097:blood; D004151:Dipeptides / Q000032:analysis* / Q000097:blood; D005978:Glutathione / Q000032:analysis* / Q000097:blood; D006710:Homocysteine / Q000032:analysis* / Q000097:blood; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D002457:Cell Extracts; D004151:Dipeptides; D006710:Homocysteine; C028505:cysteinylglycine; D005978:Glutathione; D003545:Cysteine