Development of a column-switching high-performance liquid chromatography for kynurenine enantiomers and its application to a pharmacokinetic study in rat plasma.
Kynurenine (KYN), a tryptophan metabolite, is a precursor of kynurenic acid, which is an antagonist of N-methyl-d-aspartate receptor. In this study, an enantiomeric separation of d,l-KYN derivatized with the benzofurazan fluorescence reagent 4-N,N-dimethylaminosulfonyl-7-fluoro-2,1,3-benzoxadiazole (DBD-F) (DBD-d,l-KYN) was first investigated by using a high-performance liquid chromatography (HPLC) with several chiral columns. As a consequence, DBD-d,l-KYN was enantiomerically separated on a cellulose-type chiral column (CHIRALCEL OJ-RH) with a mobile phase of H(2)O/CH(3)CN/MeOH (40/50/10) containing 0.1% acetic acid. Under this condition, the separation factor and resolution were 1.48 and 1.28, respectively. Next, a column-switching HPLC consisting of both octadecylsilica and chiral columns was developed and used to determine both d- and l-KYN enantiomers in 10 microL of rat plasma following the intraperitoneal administration of d,l-KYN to rats (10 mg kg(-1)). The result revealed that the concentration of l-KYN was higher than that of d-KYN, suggesting that d-KYN was eliminated faster than l-KYN.
Mitsuhashi
Shogo
S
Division of Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, Shizuoka 422-8526, Japan.
Fukushima
Takeshi
T
Arai
Kotaro
K
Tomiya
Masayuki
M
Santa
Tomofumi
T
Imai
Kazuhiro
K
Toyo'oka
Toshimasa
T
D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods; D002482:Cellulose / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007202:Indicators and Reagents / Q000494:pharmacology; D007263:Infusions, Parenteral / Q000379:methods; D007737:Kynurenine / Q000008:administration & dosage / Q000737:chemistry* / Q000493:pharmacokinetics*; D008297:Male; D010080:Oxazoles / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012822:Silicon Dioxide / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods; D013237:Stereoisomerism; D013449:Sulfonamides / Q000737:chemistry
D007202:Indicators and Reagents; D010080:Oxazoles; D013449:Sulfonamides; C026679:octadecylsilica; D007737:Kynurenine; D012822:Silicon Dioxide; D002482:Cellulose; C060548:4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole