An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia
An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia
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DOI：
10.1016/j.jchromb.2007.04.013
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发表时间：
2007-07-01
影响因子：
3
通讯作者：
Karnes, H. Thomas
中科院分区：
文献类型：
--
作者：
Farthing, Don;Sica, Domenic;Karnes, H. Thomas
开发了一种简单、灵敏的高效液相色谱法（利用紫外 (UV) 检测的高效液相色谱法，用于测定人血浆中的肌苷和次黄嘌呤。为了进行成分分离，使用整体式 C-18 柱，流速为 1.0 mL/min，流动相为三氟乙酸（0.11% TFA 去离子水 pH 2.2，v/v）和甲醇梯度。该方法采用离心过滤的一步样品制备方法。血浆中的成分回收率很高（接近 98%），无需使用内标，该方法对肌苷和次黄嘌呤具有出色的线性（0.25-5 μg/mL，R > 0.9990），检测限为 100 ng/mL。这种简单且经济有效的方法可用于评估潜在的内源性血浆生物标志物，可帮助医院急救人员早期发现患者的急性心脏缺血。表现为非创伤性胸痛 (C) 2007 Elsevier B.V. 保留所有权利。
A simple and sensitive high-performance liquid chromatography (HPLC method utilizing ultraviolet (UV) detection was developed for the determination of inosine and hypoxanthine in human plasma. For component separation, a monolithic C-18 column at a flow rate of 1.0 mL/min with an aqueous mobile phase of trifluoroacetic acid (0.11% TFA in deionized water pH 2.2, v/v) and methanol gradient was used. The method employed a one-step sample preparation utilizing centrifugal filtration with high component recoveries (similar to 98%) from plasma, which eliminated the need of an internal standard. The method demonstrated excellent linearity (0.25-5 mu g/mL, R > 0.9990) for both inosine and hypoxanthine with detection limits of 100 ng/mL. This simple and cost effective method was utilized to evaluate potential endogenous plasma biomarker(s), which may aid hospital emergency personnel in the early detection of acute cardiac ischemia in patients presenting with non-traumatic chest pain. (C) 2007 Elsevier B.V. All rights reserved.