Determination of L-Buthionin (SR)-Sulfoximine, g-Glutamylcysteine...

Published: Journal of Chromatographic Science, Volume 34, Number 7, July 1996, pp. 326-329.

Determination of L-Buthionin (SR)-Sulfoximine, g-Glutamylcysteine Synthetase Inhibitor in Rat Plasma with HPLC after Prelabeling with Dansyl Chloride
Hikaru Koyama, Nobuyuki Sugioka, Isao Hirata, Toshio Ohta, and Hideki Kishimoto

L-(SR)-Buthionin sulfoximine (L-(SR)-BSO) is a potent and specific inhibitor of g-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 0.01M phosphate 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.

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