甲硝唑与牛血清白蛋白相互作用的电化学行为研究.doc

甲硝唑与牛血清白蛋白相互作用的电化学行为研究 刘杰，刘艳梅，张勇 (1.太原中心医院 太原030009 2.山西大学化学化工学院，山西 太原 030006) 摘要 目的。方法循环伏安法和线性扫描伏安法。结果结论。 关键词：甲硝唑；；电化学行为Study on the interaction between metronidazole and bovine serum albumin using electrochemical method LIU Jie , ZHANG bo, (Department of pharmacentics,The Central Hospital of Taiyuan.Taiyuan 030009) LIU Yan-mei , ZHANG Yong* (School of Chemistry and Chemical Engineer, Shanxi University, Taiyuan 030006) Abstract Objective: The electrochemical methods has been used for study on the interaction between metronidazole and bovine serum albumin. Methods:Cyclic voltammetry and linear sweep voltammetry. Results: The results showed that a sensitive reduction peak of metronidazole has been obtained in the base solution of Britton-Robinson buffer solution (pH8.5) and 0.6 mol?L-1 KCl solution, but bovine serum albumin has not reduction peak in the rang of -0.3~-0.8V. When bovine serum albumin was added in metronidazole solution, the reduction peak of metronidazole will fall down. The peak current declining value was directly proportional to the bovine serum albumin concentration in the rang of 5.0×10-7~1.0×10-4 mol?L-1 (r=0.995). The detection limit was 8.0×10-8 mol?L-1. In addition, the interaction mechanism between metronidazole and BSA has been studied, the results demonstrate that combination of them mainly by hydrophobicity. Conclusion: The binding ratio and binding constant of metronidazole with BSA calculated. Furthermore , the binding mechanism was also preliminarily discussed. Key Words: metronidazole; bovine serum albumin; electrochemical behavior; interaction 蛋白质的定量测定是生物化学、药学、食品检验及临床分析中常涉及的重要内容, 目前蛋白质的定量测定主要是光谱探针法[1-8] 。近年来,电化学法由于其简便、快速、价廉, 在生命科学领域内日益受到重视[9-11] 。本文用电化学分析方法对甲硝唑和BSA之间的相互作用进行了研究。建立了一种测定BSA的新方法，并对结合反应机理进行了初步探讨[12,13]。 1 仪器与试剂 1.0×10-3 mol?L-1）： 准确称取0.0171g甲硝唑（太原制药厂，质量分数﹥99.5%），用二次蒸馏水溶解并定容于l00mL容量瓶中，置于棕色试剂瓶中于冷暗处保存；牛血清白蛋白(BSA) （Sigma公司，分子量：68000）；pH8.5的Britton-Robinson缓冲溶液；1.2 mol?L-1氯化钾溶液。(其它试剂均为分析纯，实际用水为二次蒸馏水。) 2 实验方法 10 mL 的比色管中，依次