Coudray C, Mangournet C, Bouhadjeb S, Faure H, Favier A
Laboratoire de Biochimie C, Centre hospitalo-universitaire de Grenoble, France.
J Chromatogr Sci. 1996 Apr;34(4):166-73. doi: 10.1093/chromsci/34.4.166.
The in vivo measurement of highly reactive free radicals, such as the hydroxyl radical (oOH), in humans is very difficult, if not impossible. Specific markers, based on the ability of oOH to attack aromatic molecules and produce hydroxylated compounds, are under investigation. In vivo radical metabolism of salicylic acid produces two main hydroxylated derivatives: 2,3- and 2,5-dihydroxybenzoic acid (DHBA). The measurement of 2,3-DHBA, following oral administration of salicylic acid or its acetylated form (aspirin), is proposed for the assessment of in vivo oxidative stress. The intensity of oxidative stress is a function of the ratio of dihydroxylated derivatives to salicylic acid rather than the absolute dihydroxylated derivatives levels. Consequently, a simple, accurate, and sensitive assay of the salicylic acid level in plasma is needed to investigate the in vivo free radical production. In this work, a rapid and sensitive method is presented that is useful for the quantitation of salicylic acid in biological fluids. The methodology uses high-performance liquid chromatography with spectrophotometric detection for the identification and quantitation of salicylic acid without organic extraction. A detection limit of less than 5 mumol is achieved with spectrophotometric detector responses that are linear over at least 6 orders of magnitude. Plasma concentrations of salicylic acid determined by the present technique are reported following the administration of 1000 mg aspirin in 20 healthy subjects.
在人体中对高活性自由基，如羟基自由基（·OH）进行体内测量即便并非不可能，也是非常困难的。基于·OH攻击芳香族分子并产生羟基化化合物的能力的特定标志物正在研究中。水杨酸的体内自由基代谢产生两种主要的羟基化衍生物：2,3 - 和2,5 - 二羟基苯甲酸（DHBA）。有人提出，在口服水杨酸或其乙酰化形式（阿司匹林）后测量2,3 - DHBA，以评估体内氧化应激。氧化应激的强度是二羟基化衍生物与水杨酸比值的函数，而非二羟基化衍生物的绝对水平。因此，需要一种简单、准确且灵敏的血浆水杨酸水平测定方法来研究体内自由基的产生。在这项工作中，提出了一种快速且灵敏的方法，该方法可用于定量生物流体中的水杨酸。该方法采用高效液相色谱结合分光光度检测，无需有机萃取即可鉴定和定量水杨酸。使用分光光度检测器时，检测限低于5 μmol，其响应在至少6个数量级内呈线性。在20名健康受试者服用1000 mg阿司匹林后，报告了用本技术测定的血浆水杨酸浓度。
