Open AccessSimultaneous determination of total plasma glutathione, homocysteine, cysteinylglycine, and methionine by high‐performance liquid chromatography with electrochemical detection
Author(s) -
Houze Pascal,
Gamra Stéphanie,
Madelaine Isabelle,
Bousquet Bernard,
Gourmel Bernard
Publication year - 2001
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1018
Subject(s) - homocysteine , methionine , hyperhomocysteinemia , glutathione , chemistry , chromatography , high performance liquid chromatography , thiol , medicine , plasma homocysteine , methionine synthase , endocrinology , biochemistry , amino acid , enzyme
We here describe an ion‐exchange high‐performance liquid chromatography technique with electrochemical detection for rapid quantification of glutathione, homocysteine, cysteinylglycine, and methionine. The analytical validation of the technique showed within‐assay and between‐assay coefficients of variation between 3.1 and 4.3%, and 3.7 and 8.6%, respectively. Percentages of recovery for overload and dilution tests were between 87 and 120%. Detection limits were 1 μmol/L for methionine and 0.5 μmol/L for other compounds. There was no interference with any physiological and pharmacological substances possessing a thiol function. Aminothiol concentrations determined in 100 control subjects (50 women and 50 men) showed no age‐ or sex‐rated differences for except for homocysteine which was increased (+ 28%) in oldest subjects of both sexes. In 60 patients at risk (30 with chronic renal failure, 30 with diabetes), homocysteine concentration was significantly increased. No variation in other aminothiols was observed in diabetic subjects. Methionine was decreased and cysteinylglycine was increased in patients with chronic renal failure. The present technique—rapid, easy to use, and reliable—appears suitable for routine application in the exploration of aminothiol metabolic pathways including mechanisms of hyperhomocysteinemia. J. Clin. Lab. Anal. 15:144–153, 2001. © 2001 Wiley‐Liss, Inc.