Glyoxylic acid (GA) is the intermediate metabolite in various mammalian metabolic pathways. GA showed high reactivity towards formation of advanced glycation end-products (AGEs); the main cause of pathogenesis and complications of many diseases. The presented study aimed to detect GA in healthy and cardiovascular patients' (CV) sera; however analysis of GA in biological fluid is a challenge and requires chemical derivatization. Hence, a new, highly sensitive, time saving and reproducible pre-column fluorescence derivatization procedure coupled with high performance liquid chromatography (HPLC) method was developed. The derivatization method was based on reaction of 2-aminobenzenthiol (2-ABT), a fluorogenic reagent, with GA in acidic medium to form highly fluorescent thiazine derivative (290 and 390 nm for excitation and emission wavelengths respectively). The fluorescent derivative was separated within 6 min on a reversed-phase ODS column using an isocratic elution with a mixture of methanol-water (70:30, v/v%). The proposed method parameters were optimized and the method was validated. A good linearity in the concentration range (0.05-5.0 µM) was obtained with detection limit (LOD) of 10 nM (200 fmol/injection), which is more sensitive than several previous methods. Moreover, the recovery results were within the range of 85.0-95.5 % and the intra- and inter-day precision results were ≤3.5%. It should be emphasized that this method is the first one for monitoring of GA in CV patients; to investigate its role for diagnosis and monitoring the severity and complications of this disease in clinical laboratory.

中文翻译：

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基于噻嗪衍生物形成的柱前荧光衍生法估算乙醛酸的HPLC方法的开发：在健康和心血管患者血清中的新应用。

乙醛酸（GA）是各种哺乳动物代谢途径中的中间代谢产物。GA对高级糖基化终产物（AGEs）的形成具有高反应性；多种疾病的发病机理和并发症的主要原因。本研究旨在检测健康和心血管患者（CV）血清中的GA；然而，分析生物流体中的GA是一项挑战，需要化学衍生化。因此，开发了一种新的，高度灵敏，省时且可重现的柱前荧光衍生化方法，并结合了高效液相色谱（HPLC）方法。衍生方法基于发荧光试剂2-氨基苯硫醇（2-ABT）的反应，用GA在酸性介质中形成高度荧光的噻嗪衍生物（激发和发射波长分别为290和390 nm）。在反相ODS色谱柱上使用甲醇-水（70：30，v / v％）的混合物进行等度洗脱，在6分钟内分离出荧光衍生物。对提出的方法参数进行了优化，并对方法进行了验证。在检测限（LOD）为10 nM（200 fmol /进样）的条件下，浓度范围（0.05-5.0 µM）处具有良好的线性，这比以前的几种方法更为灵敏。此外，回收率结果在85.0-95.5％的范围内，日内和日间精度结果≤3.5％。应该强调的是，这种方法是监测CV患者GA的第一种方法。