用反相高效液相色谱法分析腺苷及其在腺苷激酶动力学研究中的应用。

Analysis of adenosine by RP-HPLC method and its application to the study of adenosine kinase kinetics.

作者信息

Marin Rodrigo Miguel, Franchini Kleber Gomes, Rocco Silvana Aparecida

机构信息

Department of Internal Medicine, School of Medicine, UNICAMP, Campinas, SP, Brazil.

出版信息

J Sep Sci. 2007 Oct;30(15):2473-9. doi: 10.1002/jssc.200700194.

DOI:10.1002/jssc.200700194

PMID:17763527

Abstract

An RP-HPLC method for the analysis of adenosine (ADO) has been developed and validated. In the present study, we report an RP-HPLC-based method with modifications of mobile phase and shorter retention time that substantially improved the efficiency of ADO analysis. The HPLC separation of the ADO was achieved on a C18 column, using a mobile phase consisting of water, containing 7% v/v ACN, at a flow rate of 0.8 mL/min. The column effluent was monitored by UV detection at 260 nm. A linear response was achieved over the concentration range of 0.25-100.00 micromol/L. The analytical method inter- and intra-run accuracy and precision were better than +/- 15%. The LOQ was 0.25 micromol/L, with ADO detection in the range of 6.25 pmol per sample. The method has been applied to the study of adenosine kinase (AK) kinetics.

摘要

已开发并验证了一种用于分析腺苷（ADO）的反相高效液相色谱（RP-HPLC）方法。在本研究中，我们报告了一种基于RP-HPLC的方法，该方法对流动相进行了改进，保留时间更短，大大提高了ADO分析的效率。在C18柱上实现了ADO的HPLC分离，使用由含有7%（v/v）乙腈的水组成的流动相，流速为0.8 mL/min。通过在260 nm处的紫外检测监测柱流出物。在0.25-100.00 μmol/L的浓度范围内实现了线性响应。分析方法的批间和批内准确度和精密度均优于±15%。定量限为0.25 μmol/L，每个样品中ADO的检测范围为6.25 pmol。该方法已应用于腺苷激酶（AK）动力学的研究。