The assay of S-D-lactoylglutathione in biological systems.
A procedure for the assay of S-D-lactoylglutathione, the physiological intermediate of the glyoxalase system, in biological systems is described, together with sample storage, sample processing, and statistical evaluation. Specimen data are presented. S-D-Lactoylglutathione was assayed by reverse-phase high-performance liquid chromatography (HPLC) with spectrophotometric detection of the thiolester chromophore at 233 nm. The biological sample was deproteinized with perchloric acid and partially purified by strong anion-exchange solid-phase extraction prior to HPLC. The limit of detection was 3.7 nmol, the recovery 49 +/- 4%, and the intra- and interbatch coefficients of variance 0.7 and 12%, respectively. The concentration of S-D-lactoylglutathione in whole blood from normal control human subjects was (mean +/- SD, nmol/ml whole blood) 16.5 +/- 4.4 (n = 8), and from diabetic patients 21.2 +/- 9.2 (n = 25), which is a significant increase (P < 0.05) from normal controls. The assay of S-D-lactoylglutathione is of increasing interest in studies of the elevation of glyoxalase metabolites in diabetes mellitus and in investigations of the antiproliferative activity of S-D-lactoylglutathione.
McLellan
A C
AC
Department of Chemistry and Biological Chemistry, University of Essex, Colchester, United Kingdom.
Phillips
S A
SA
Thornalley
P J
PJ
D000818:Animals; D016477:Artifacts; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003920:Diabetes Mellitus / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives* / Q000032:analysis; D006801:Humans; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D014407:Tumor Cells, Cultured
D005978:Glutathione; C013585:S-lactoylglutathione