title: Determination of rat blood S-D-lactoylglutathione by a column-switching high-performance liquid chromatography with precolumn fluorescence derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole.
abstract: A method for the determination of S-d-lactoylglutathione (SLG), an intermediate metabolite of the glyoxalase system, in rat blood is described. After hemolysis and deproteinization of 30 microl of rat blood, SLG in the blood was determined by a column-switching HPLC system with precolumn fluorescence derivatization with a fluorogenic reagent, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). Calibration curves for the determination of SLG showed a good linearity (r2 > 0.999) over the range of 20-100 pmol SLG spiked in rat blood samples. The accuracy was in the range of 97-104% (20-100 pmol SLG spiked in rat blood sample, N = 4). The detection limit was 8.12 fmol, and the quantitation limit was 27.07 fmol, respectively. The intra- and interday coefficients of variance were 4.63% (N = 5) and 9.98% (N = 5), respectively. The concentrations of SLG in whole blood from male Wistar-Kyoto rats (12 weeks old) were 3.48+/-0.78 microM (N = 4, mean+/-SE). In streptozotocin-induced diabetic rats, the concentration of SLG was significantly increased, approx 5-fold, compared with normal rats, suggesting that the metabolic flux of the glyoxalase system increases in red blood cells during hyperglycemia.
authors.affiliation: Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Erika
authors.initials: E
authors.lastname: Uchino
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fukushima
authors.forename: Makoto
authors.initials: M
authors.lastname: Tsunoda
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Santa
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Imai
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D003921:Diabetes Mellitus, Experimental / Q000175:diagnosis; D005456:Fluorescent Dyes; D005978:Glutathione / Q000031:analogs & derivatives* / Q000097:blood*; D006943:Hyperglycemia / Q000175:diagnosis; D008297:Male; D051381:Rats; D011921:Rats, Inbred WKY; D015203:Reproducibility of Results; D013311:Streptozocin
chemical_list: D005456:Fluorescent Dyes; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; D013311:Streptozocin; D009327:4-Chloro-7-nitrobenzofurazan; D005978:Glutathione; C013585:S-lactoylglutathione