Determination of L-Buthionin (SR)-Sulfoximine, -Glutamylcysteine Synthetase Inhibitor in Rat Plasma with HPLC after Prelabeling with Dansyl Chloride

H. Koyama, N. Sugioka, I. Hirata, T. Ohta, H. Kishimoto
1996 Journal of Chromatographic Science  
L-(SR)-Buthionin sulfoximine (L-(SR)-BSO) is a potent and specific inhibitor of γ-glutamylcysteine synthetase, which catalyzes the first reaction of glutathione biosynthesis. A selective, sensitive, and simple high-performance liquid chromatographic method was developed for the determination of L-(SR)-BSO in rat plasma. After the compound was labeled with dansyl chloride (Dns-Cl) under optimal conditions, it was separated in a Zolbax-ODS column with a mobile phase that consisted of 0.01 Μ
more » ... ate buffer, methanol, and acetonitrile (8:1:3, v/v). The compound was detected with a fluorescence detector at an excitation wavelength of 335 nm and an emission wavelength of 525 nm using a xenon lamp. The coefficients of variation (CV) from the interassay in the low and high concentrations (10 and 500 μg/mL of L-(SR)-BSO in rat plasma) were 2.5 and 4.8%, respectively. The CVs from the intra-assay in the low and high concentrations were 3.2 and 5.6%, respectively. The minimum concentration of L-(SR)-BSO that could be determined was 10 μg/mL when 100-μL serum samples were used. The detection limit was 50 ng per injection volume. This method enables pharmacokinetic and pharmacodynamic studies in rats.
doi:10.1093/chromsci/34.7.326 pmid:8675550 fatcat:lakncdzeuzfgphh6fedxuux6yq