Yao T, Matsumoto Y, Wasa T
Department of Applied Chemistry, College of Engineering, University of Osaka Prefecture, Sakai, Japan.
J Biotechnol. 1990 Apr;14(1):89-97. doi: 10.1016/0168-1656(90)90021-3.
A sensitive and highly selective method for the simultaneous determination of purine bases and their nucleosides is proposed. An amperometric flow-injection system with the two immobilized enzyme reactors (guanase immobilized reactor and purine nucleoside phosphorylase/xanthine oxidase co-immobilized reactor) is used as the specific post-column detection system of HPLC, to convert compounds separated by a reversed-phase. HPLC column to electroactive species (hydrogen peroxide and uric acid) which can be detected at a flow-through platinum electrode. The proposed detection system is specific for a group of purine bases and purine nucleosides and does not respond for purine nucleotides and pyrimidine bases. The linear determination ranges are from 10 pmol to 5 nmol for four purine bases (hypoxanthine, xanthine, guanine, and adenine) and four purine nucleosides (inosine, xanthosine, guanosine, and adenosine). The detection limits are 1.2-5.5 pmol.
提出了一种同时测定嘌呤碱及其核苷的灵敏且高选择性方法。采用带有两个固定化酶反应器（鸟嘌呤酶固定化反应器和嘌呤核苷磷酸化酶/黄嘌呤氧化酶共固定化反应器）的安培流动注射系统作为高效液相色谱（HPLC）的特定柱后检测系统，将通过反相HPLC柱分离的化合物转化为可在流通式铂电极上检测的电活性物质（过氧化氢和尿酸）。所提出的检测系统对一组嘌呤碱和嘌呤核苷具有特异性，对嘌呤核苷酸和嘧啶碱无响应。四种嘌呤碱（次黄嘌呤、黄嘌呤、鸟嘌呤和腺嘌呤）和四种嘌呤核苷（肌苷、黄苷、鸟苷和腺苷）的线性测定范围为10皮摩尔至5纳摩尔。检测限为1.2 - 5.5皮摩尔。
