A high‐performance liquid chromatography assay with a triazole‐bonded column for evaluation of d‐amino acid oxidase activity. (27th August 2015)

• Record Type: Journal Article
• Authors: Iwasaki, Megumi; Kashiwaguma, Yoshiyuki; Nagashima, Chihiro; Izumi, Mao; Uekusa, Ayano; Iwasa, Sumiko; Onozato, Mayu; Ichiba, Hideaki; Fukushima, Takeshi
• Is Part Of: Biomedical chromatography. Volume 30:Number 3(2016:Mar.)
• Page Start: 384
• Page End: 389
• Publication Date: 2015-08-27
• Subjects: d‐amino acid oxidase -- kynurenic acid -- d‐kynurenine -- triazole‐bonded stationary phase -- Cosmosil HILIC®; Chromatographic analysis -- Periodicals; Biology -- Periodicals; Medicine -- Periodicals; Biology -- Periodicals; Chromatography -- methods -- Periodicals; Medicine -- Periodicals; 543.089
• DOI: 10.1002/bmc.3559
• Languages: English
• ISSNs: 0269-3879

Abstract

Abstract: Elution profiles of kynurenic acid (KYNA) and 7‐chlorokynurenic acid (Cl‐KYNA) were examined by high‐performance liquid chromatography (HPLC) using a triazole‐bonded stationary phase column (Cosmosil® HILIC) under isocratic elution of a mobile phase consisting of CH3 CN–aqueous 10 mm ammonium formate between pH 3.0 and 6.0. The capacity factors of KYNA and Cl‐KYNA varied with both the CH3 CN content and the pH of the mobile phase. The elution order of KYNA and Cl‐KYNA was reversed between the CH3 CN‐ and H2 O‐rich mobile phases, suggesting that hydrophilic interactions and anion‐exchange interactions caused retention of KYNA and Cl‐KYNA in the CH3 CN‐ and H2 O‐rich mobile phases, respectively. The present HPLC method using a triazole‐bonded column and fluorescence detection (excitation 250 nm, emission 398 nm) was applied to monitor in vitro production of KYNA fromd ‐kynurenine (d ‐KYN) byd ‐amino acid oxidase (DAO) using Cl‐KYNA as an internal standard. A single KYNA peak was clearly observed after enzymatic reaction ofd ‐KYN with DAO. Production of KYNA fromd ‐KYN was suppressed by the addition of commercial DAO inhibitors. The present HPLC method can be used to evaluate DAO activity and DAO inhibitory effects in candidate drugs for the treatment of schizophrenia. Copyright © 2015 John Wiley & Sons, Ltd.