Faculty of Pharmacy, Iwaki Meisei University, Iwaki, Fukushima 970-8551, Japan.
J Fluoresc. 2011 Nov;21(6):2093-100. doi: 10.1007/s10895-011-0908-1. Epub 2011 Jun 7.
It was shown that ubiquinone (CoQ(10)) and ubiquinol (CoQ(10)H(2)) produce fluorescence products under alkaline conditions when reacted with 2-cyanoacetamide. The reaction mixture from CoQ(10) gave fluorescence with excitation and emission maximum wavelengths at 442 nm and 549 nm, respectively. This reaction was considered to proceed via Craven's reaction. Moreover, 2-cyanoacetamide was shown to be a useful reagent for high-performance liquid chromatography (HPLC) with post-column fluorescence derivatization of CoQ(10) and CoQ(10)H(2) in blood. CoQ(10) showed a linear response in the range of 0.32-1276 ng, and the detection limit (S/N = 3) was 0.16 ng. Moreover, the sample pretreatment by deproteinization and extraction of CoQ(10) and CoQ(10)H(2) from plasma using 1-propanol with potassium formate was effective for excellent separation of CoQ(10) and CoQ(10)H(2) from other fluorescent substances in the blood. This simple and rapid pretreatment was considered to minimize the oxidation of CoQ(10)H(2). On the other hand, CoQ(10) and CoQ(10)H(2) in plasma samples obtained by finger prick were detected, as in venous blood obtained by venipuncture. Our method involving the simple and rapid collection of plasma by finger prick and sample pretreatment is thought to be applicable for the determination of CoQ(10)H(2)/total CoQ(10) ratio as a biomarker of oxidative stress.
研究表明，当泛醌（CoQ（10））和泛醇（CoQ（10）H（2））在碱性条件下与 2-氰基乙酰胺反应时，会产生荧光产物。CoQ（10）的反应混合物在激发和发射最大波长分别为 442nm 和 549nm 时具有荧光。该反应被认为是通过 Craven 反应进行的。此外，2-氰基乙酰胺被证明是一种有用的试剂，可用于高效液相色谱（HPLC），可对血液中的 CoQ（10）和 CoQ（10）H（2）进行柱后荧光衍生化。CoQ（10）在 0.32-1276ng 的范围内呈现线性响应，检测限（S/N=3）为 0.16ng。此外，通过用甲酸钾的 1-丙醇对 CoQ（10）和 CoQ（10）H（2）进行蛋白沉淀和提取，对血浆进行样品预处理，对于从血液中的其他荧光物质中极好地分离 CoQ（10）和 CoQ（10）H（2）是有效的。这种简单快速的预处理被认为可以最大程度地减少 CoQ（10）H（2）的氧化。另一方面，通过手指穿刺获得的血浆中的 CoQ（10）和 CoQ（10）H（2）与通过静脉穿刺获得的静脉血中的 CoQ（10）和 CoQ（10）H（2）一样被检测到。我们的方法涉及通过手指穿刺简单快速地采集血浆并进行样品预处理，被认为适用于测定 CoQ（10）H（2）/总 CoQ（10）比作为氧化应激的生物标志物。
