柱前衍生化HPLC用于顺铂纳米粒大鼠药动学的研究  

A modified precolumn derivatization high-performance liquid chromatography for pharmacok inetics study of cisplatin nanoparticles in rats

作　　者：朱珊珊[1] 宋娟[1] 任伟芳 王姝[1] 张易[1] 郭宏宇[1] 杨丽[1] 

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2017年第2期158-163,共6页Journal of Shenyang Pharmaceutical University

摘　　要：目的建立一种改良的柱前衍生化-高效液相色谱法(HPLC)测定顺铂纳米粒大鼠血浆药物浓度,并将其应用于大鼠体内药动学研究。方法采用衍生化试剂二乙基二硫代氨基甲酸钠(DDTC)与顺铂络合生成Pt(DDTC)2衍生化产物,加入内标物地西泮,氯仿提取,挥干,乙酸乙酯复溶,采用HPLC法测定。色谱条件:色谱柱为C18,流动相为水-甲醇(体积比为1∶3),检测波长为252 nm。大鼠以7.5 mg·kg-1剂量分别尾静脉注射顺铂纳米粒和注射剂,不同时间取血,采用所建立的分析方法测定血浆顺铂质量浓度,计算药动学参数。结果本法可精密准确地测定顺铂纳米粒静脉注射给药后大鼠血浆中的顺铂质量浓度,线性范围为0.488~97.6 mg·L^(-1),最低定量限为100μg·L^(-1),日内、日间变异系数均<5%。顺铂纳米粒的主要药动学参数:ρmax为(62.85±13.83)mg·L^(-1),t1/2为(3.53±1.57)min,AUC0-∞为(176.81±46.19)mg·L^(-1)·h,CL为(44.89±11.91)m L·kg·h-1。结论该方法适用于顺铂纳米粒体内药物质量浓度测定及其药动学的研究。与顺铂注射液相比,顺铂纳米粒药时曲线下面积显著提高,清除率显著降低,其药动学的改变有利于降低药物的毒副作用。Objective To develop and validate a modified precolumn derivatization high-performance liquid chromatography（ HPLC） method for the determination of cisplatin（ CDDP） in rats plasma and for pharmacokinetics study. Methods CDDP was measured in plasma following derivatization with sodium diethyldithioearbamate（ DDTC） by quantization against diazepam as internal standard. After addition of diazepam,the mixture was extracted with chloroform by vortexing. Then the chloroform layer was evaporated to dryness with nitrogen gas and the residue was redissolved with ethyl acetate. Subsequently,an ethyl acetate containing Pt（ DDTC）2and diazepam was separated by HPLC on a C18 column using watermethanol（ V / V = 1 ∶ 3） as the mobile phase. Detection was carried out by absorbance at 252 nm. Rats were injected intravenous with CDDP-S or CDDP-N（7. 5 mg·kg^- 1）. For drug concentration assays,samples were collected at different times after the injection. Ultimately,the CDDP concentration was determined with validated precolumn derivatization HPLC method,and the pharmacokinetic parameters were estimated.Results The method proved to be accurate and effective for measuring the concentrations of CDDP. The method was linear in the range of 0. 488- 97. 6 mg·L^-1,and the limit quantification was 100 μg·L^-1.Moreover,the coefficients variation of inter-day and intra-day were both less than 5%. Mainly pharmacokinetic parameters of CDDP-N were calculated as follow：ρmax=（62. 85 ± 13. 83） mg·L^-1,t1 /2=（3. 53 ± 1. 57） min,AUC0-∞=（ 176. 81 ± 46. 19） mg·L^-1·h,CL =（44. 89 ± 11. 91） m L·kg·h^- 1.Conclusions The method described in the present study appears to possess a high degree of specificity,precision and economy for determination of CDDP in rat plasma and pharmacokinetics study. CDDP-N exhibited a higher AUC and a lower CL compared to CDDP-S,which is beneficial to decrease toxicity of CDDP in vivo.

关 键 词：顺铂 柱前衍生化 高效液相色谱法 二乙基二硫代氨基甲酸钠 纳米粒 药动学 

分 类 号：R94[医药卫生—药剂学]