高效液相色谱法测定血清中cyp3a4探针药物氨苯砜浓度.doc

高效液相色谱法测定血清中CYP3A4探针药物氨苯砜浓度 谢建芬1, 2*，陈荣2，夏宗玲2，丁选胜1# （1.中国药科大学临床药学教研室，南京210009；2.江苏常州市第一人民医院药剂科，常州市 213003） [摘要] 目的：建立高效液相色谱法测定细胞色素P450 3A4（CYP3A4）探针药物氨苯砜的血药浓度。方法：采用迪马Diamonsil C18色谱柱（250×4.6mm,5μm）；流动相：乙腈-水（25:75 V/V）；流速：1.0mL?min-1；紫外检测波长：258nm；柱温：30℃；进样：30μL。结果：氨苯砜在1~6mg?L-1 范围内线性关系良好，r=0.9991。日内RSD≤7.05%,日间RSD≤5.54%。结论：本法操作简便，灵敏度高，快速可靠，适用于患者体内氨苯砜含量的测定。 [关键词]：HPLC；CYP3A4；探针药物；氨苯砜 Determination of Cytochrome P450 3A4 Probe Drug Dapsone in serum using High Performance Liquid Chromatography XIE Jian-fen, CHEN Rong, XIA Zong-ling, DING Xuan-sheng(1.Department of Clinical Pharmacy, China Pharmaceutical University, Nanjing 210009, Chian; 2.Department of Pharmacy, First People’s Hospital of Changzhou, Changzhou 213003, China) ABSTRACT：OBJECTIVE To establish the method for determination of cytochrome P450 3A4 probe drug dapsone in plasma by HPLC. METHODS The chromatographic separation dapsone from serum were achieved using C18(250×4.6mm,5μm ) column and the mobile phase was acetonitrile-water (25:75 v/v ), and flow rate was 1.0 mL?min-1. They were detected at 258nm wave lenth, and the column temperature was 30℃, the sampling volume was 30μL. RESULTS Excellent linearity was observed during the range of 1mg?L-1 to 6mg?L-1 of dapsone in plasma (r=0.9991). The RSD within a day and between days were less than 7.05% and 5.54%. CONCLUSION The high-performance liquid chromatographic method is simple, accurate, stable and suitable for measuring dapsone in patient. Key words: HPLC; CYP3A4; Probe drug; Dapsone 细胞色素P450（CYP450）为肝脏微粒体混合功能氧化酶系的主要成分，其实际上为同一家庭的多种异构型，在药理学研究和药物代谢中有重要意义。基本上分成至少4个基因族，分别为CYP1，CYP2，CYP3和CYP4[1]。其中CYP1，2，3，主要代谢外源性化合物，如药物、毒物等，有交叉的底物特异性，常可被外源性物质诱导。CYP450的活性可以通过一些特异性敏感底物来测定，相关的研究也较多[2-3]，其中CYP3A常用的探针药物有氨苯砜、红霉素、咪达唑仑等[4]。Sharma Ashish等[5]用HPLC内标法检测了健康者血浆中美托洛尔、氯唑沙宗、甲苯磺丁脲、氨苯砜和咖啡因5种探针药物的含量。本实验在文献的基础上，对CYP3A4探针药物氨苯砜的测定方法进行改进，以达到快速、准确、灵敏度高、专属性强。 Waters高效液相色谱系统包括2996紫外检测器、1525双泵、717自动进样器、Empower色谱工作站；电子分析天平（德国塞多利斯仪器有限公司，型号：BT25S，精密度1×10-5g）；高速台式离心机（科大创新股份有限公司中佳分公司，型号：KD