从5'-单磷酸胞苷生物合成的5'-三磷酸胞苷在离子交换树脂上的分离及胞苷化合物的高效液相色谱分析

Separation of cytidine 5'-triphosphate biosynthesized from cytidine 5'-monophosphate on ion-exchange resin and HPLC analysis of cytidine compounds.

作者信息

Shi Lu-E, Ying Guo-Qing, Tang Zhen-Xing, Yi Yu, Shan Jian-Feng, Liu Hua-Zhang

机构信息

College of Chemical Engineering and Materials Science, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.

出版信息

Appl Biochem Biotechnol. 2008 Jan;144(1):1-14. doi: 10.1007/s12010-007-8015-0.

DOI:10.1007/s12010-007-8015-0

PMID:18415982

Abstract

Conditions were studied in the biosynthesis of cytidine 5'-triphosphate (CTP) from cytidine 5'-monophosphate (CMP). A 201 x 7 anion ion-exchange resin was applied for the separation of CTP from CMP. Adsorption isotherm and elution conditions (eluant, eluant concentration, flow rate, sample volume loaded) were investigated. At the same time, a new high-performance liquid chromatography on an anion ion-exchange column WAX-1 with UV detector at 260 nm was developed to measure CMP, cytidine 5'-diphosphate (CDP), and CTP. The retention time for CMP, CDP, and CTP are 0.723, 1.448, and 4.432 min, respectively. This new rapid high-performance liquid chromatography (HPLC) method for the analysis of cytidine compounds in biological sample has a wide linear range with high precision and repeatability.

摘要

对从胞苷 5'-单磷酸（CMP）生物合成胞苷 5'-三磷酸（CTP）的条件进行了研究。使用 201 x 7 阴离子离子交换树脂从 CMP 中分离 CTP。研究了吸附等温线和洗脱条件（洗脱剂、洗脱剂浓度、流速、上样样品体积）。同时，开发了一种在配备 260 nm 紫外检测器的阴离子离子交换柱 WAX-1 上的新型高效液相色谱法，用于测定 CMP、胞苷 5'-二磷酸（CDP）和 CTP。CMP、CDP 和 CTP 的保留时间分别为 0.723、1.448 和 4.432 分钟。这种用于分析生物样品中胞苷化合物的新型快速高效液相色谱（HPLC）方法具有宽线性范围，且具有高精度和可重复性。