使用含18-冠-6流动相的高效液相色谱光照射系统同时测定犬尿氨酸和犬尿喹啉酸

Simultaneous Determination of Kynurenine and Kynurenic Acid by High-Performance Liquid Chromatography Photoirradiation System Using a Mobile Phase Containing 18-crown-6.

作者信息

Atsumi Motomasa, Mawatari Ken-Ichi, Morooka Akari, Yasuda Makoto, Fukuuchi Tomoko, Yamaoka Noriko, Kaneko Kiyoko, Nakagomi Kazuya, Oku Naoto

机构信息

Faculty of Pharma-Science, Teikyo University, Tokyo, Japan.

出版信息

Int J Tryptophan Res. 2019 Mar 14;12:1178646919834551. doi: 10.1177/1178646919834551. eCollection 2019.

A high-performance liquid chromatography (HPLC) system has been developed for the fluorometric determination of kynurenine (KYN) and kynurenic acid (KYNA) in human serum using a mobile phase containing 18-crown-6. A retention time of KYNA was adjusted with pH of phosphate buffer in 18-crown-6. KYN and KYNA were separated on a CAPCELLPAK C18 (250 × 4.6 mm i.d.). The mobile phase consisted of 35 mmol/L phosphate buffer (pH 8.0)/methanol (85/15, v/v) containing 35 mmol/L hydrogen peroxide and 10 mmol/L 18-crown-6. The retention times of KYN and KYNA were 18and 24 minutes, respectively. The calibration graphs of KYN and KYNA were linear over the range 180 to 2900 and 1 to 84 nmol/L by injecting a 50-μL volume of KYN and KYNA, respectively. Pretreatment of serum was achieved by deproteinization only. The mean recoveries of KYN and KYNA from serum were more than 97%.

已开发出一种高效液相色谱（HPLC）系统，用于使用含有18-冠-6的流动相荧光法测定人血清中的犬尿氨酸（KYN）和犬尿酸（KYNA）。通过18-冠-6中磷酸盐缓冲液的pH值调节KYNA的保留时间。KYN和KYNA在CAPCELLPAK C18（250×4.6 mm内径）上分离。流动相由含有35 mmol/L过氧化氢和10 mmol/L 18-冠-6的35 mmol/L磷酸盐缓冲液（pH 8.0）/甲醇（85/15，v/v）组成。KYN和KYNA的保留时间分别为18分钟和24分钟。通过分别注入50 μL体积的KYN和KYNA，KYN和KYNA的校准曲线在180至2900和1至84 nmol/L范围内呈线性。血清预处理仅通过脱蛋白实现。血清中KYN和KYNA的平均回收率超过97%。