Abstract |
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.
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Authors | A C McLellan, S A Phillips, P J Thornalley |
Journal | Analytical biochemistry
(Anal Biochem)
Vol. 211
Issue 1
Pg. 37-43
(May 15 1993)
ISSN: 0003-2697 [Print] United States |
PMID | 8323036
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Glutathione
- S-lactoylglutathione
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Topics |
- Animals
- Artifacts
- Calibration
- Chromatography, High Pressure Liquid
(methods)
- Diabetes Mellitus
(metabolism)
- Glutathione
(analogs & derivatives, analysis)
- Humans
- Male
- Rats
- Rats, Wistar
- Reproducibility of Results
- Sensitivity and Specificity
- Spectrophotometry, Ultraviolet
- Tumor Cells, Cultured
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