Chemiluminescence assay for quinones based on generation of reactive oxygen species through the redox cycle of quinone
Chemiluminescence assay for quinones based on generation of reactive oxygen species through the redox cycle of quinone
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DOI：
10.1007/s00216-008-2541-7
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发表时间：
2009-02-01
影响因子：
4.3
通讯作者：
Kuroda, Naotaka
中科院分区：
文献类型：
--
作者：
Kishikawa, Naoya;Ohkubo, Nobuhiro;Kuroda, Naotaka
建立了一种灵敏、选择性的化学发光法测定醌类化合物。该方法以醌与二硫苏糖醇为还原剂，通过氧化还原反应生成活性氧为基础，再用发光胺化学发光法检测生成的活性氧。化学发光强度大，寿命长，且与醌浓度成正比。结果表明，超氧化物阴离子参与了化学发光反应，因为只有超氧化物歧化酶存在时，化学发光强度才会降低。在所测试的醌类中，9,10-菲蒽醌、1,2-萘醌和1,4-萘醌均有化学发光，而9,10-蒽醌和1,4-苯醌均无化学发光。醌类化合物的结构不同，其化学发光性质也有很大差异。化学发光也被观察到对生物学上重要的醌，如泛醌。因此，建立了一种基于化学发光反应的快速检测药物制剂中泛醌的方法。泛醌的检出限（空白+ 3SD）为0.05 μ M （9 ng/次），分析时间为30 s /个样品。该方法可直接测定药物制剂中的泛醌，无需任何纯化程序。
A sensitive and selective chemiluminescence assay for the determination of quinones was developed. The method was based on generation of reactive oxygen species through the redox reaction between quinone and dithiothreitol as reductant, and then the generated reactive oxygen was detected by luminol chemiluminescence. The chemiluminescence was intense, long-lived, and proportional to quinone concentration. It is concluded that superoxide anion was involved in the proposed chemiluminescence reaction because the chemiluminescence intensity was decreased only in the presence of superoxide dismutase. Among the tested quinones, the chemiluminescence was observed from 9,10-phenanthrenequinone, 1,2-naphthoquinone, and 1,4-naphthoquinone, whereas it was not observed from 9,10-anthraquinone and 1,4-benzoquinone. The chemiluminescence property was greatly different according to the structure of quinones. The chemiluminescence was also observed for biologically important quinones such as ubiquinone. Therefore, a simple and rapid assay for ubiquinone in pharmaceutical preparation was developed based on the proposed chemiluminescence reaction. The detection limit (blank + 3SD) of ubiquinone was 0.05 mu M (9 ng/assay) with an analysis time of 30 s per sample. The developed assay allowed the direct determination of ubiquinone in pharmaceutical preparation without any purification procedure.