一种用于测定健康志愿者和潜在急性心脏缺血患者人血浆中肌苷和次黄嘌呤的高效液相色谱法。
An HPLC method for determination of inosine and hypoxanthine in human plasma from healthy volunteers and patients presenting with potential acute cardiac ischemia.
作者信息
Farthing Don, Sica Domenic, Gehr Todd, Wilson Bill, Fakhry Itaf, Larus Terri, Farthing Christine, Karnes H Thomas
机构信息
Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, VCU Medical Center, Virginia Commonwealth University, Richmond, VA 23298, USA.
出版信息
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jul 1;854(1-2):158-64. doi: 10.1016/j.jchromb.2007.04.013. Epub 2007 Apr 18.
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 (0.1% 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 (approximately 98%) from plasma, which eliminated the need of an internal standard. The method demonstrated excellent linearity (0.25-5 microg/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.
建立了一种利用紫外（UV）检测的简单灵敏的高效液相色谱（HPLC）方法，用于测定人血浆中的肌苷和次黄嘌呤。为了进行成分分离，使用了整体式C（18）柱，流速为1.0 mL/min，流动相为含三氟乙酸（0.1% TFA于去离子水，pH 2.2，v/v）和甲醇的水相梯度。该方法采用一步样品制备，通过离心过滤从血浆中获得较高的成分回收率（约98%），无需内标。该方法对肌苷和次黄嘌呤均显示出优异的线性（0.25 - 5 μg/mL，R>0.9990），检测限为100 ng/mL。这种简单且经济高效的方法用于评估潜在的内源性血浆生物标志物，这可能有助于医院急救人员早期检测非创伤性胸痛患者的急性心肌缺血。
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