酶解-HPLC法测定茴三硫代谢物及人体药代动力学研究

目的: 用酶解法解离茴三硫体内代谢物对羟基苯基三硫酮,建立HPLC方法检测其含量,研究茴三硫体内代谢物对羟基苯基三硫酮在人体内的药代动力学,并评价茴三硫片剂的生物等效性。 方法: 用β-葡糖苷酸酶酶解脱去葡萄糖,使血浆中茴三硫代谢物对羟基苯基三硫酮游离后用HPLC-UV法测定其血浆中浓度;固定相:C₁₈色谱柱,流动相:甲醇-水(80∶ 20,v/v),流速:1.0 mL·min^-1,柱温:30 ℃,检测波长:346 nm,进样量:20 μL。运用DAS 2.0软件处理血药浓度数据和计算参数,并进行统计学分析。 结果: 建立的HPLC法测定血浆中对羟基苯基三硫酮浓度,专属性强,线性范围为20~1500 ng·mL^-1, r=0.9992,最低检测限为10 ng·mL^-1;提取回收率为32.2%~44.0%,方法回收率为102.5%~107.0%,日内、日间RSD均小于9%。 结论: 本项研究所用方法对羟基苯基三硫酮血药浓度检测灵敏度高、特异性强、重复性好,测定结果可靠,统计学分析表明2种制剂的主要药代动力学参数无显著性差异,具生物等效性。

Objective: To establish HPLC method to detect the metabolite of anethol trithione (4-hydroxy-anethol trithione,ATX) in vivo after zymolysis and research the pharmacokinetics,evaluate the bioequivalence of two different anethol trithione tablets. Methods: The method entailed enzymatic hydrolysis of human plasma samples via β-glucuronidase,ATX concentration was determined by HPLC with a reverse phase C₁₈ column,mobile phase consisted of a mixture of methanol-water (80∶ 20) was delivered at a flow rate of 1.0 mL·min^-1.The injection volume was 20 μL and the detection was performed at a wavelength of 346 nm.The pharmacokinetics parameters and relative bioavailability were calculated on the basis of compartment models by using DAS 2.0 program to evaluate the bioequivalence of two preparations. Results: The method of HPLC was effectively in determining 4-hydroxy-anethole trithione ATX in human plasma.It was validated and proved to be linear in the range of 20-1500 ng·mL^-1;and the limit of quantification concentration (LOQ) in plasma was 10 ng·mL^-1.The estration recovery was 32.2%-44.0% and the method recovery was 102.5%-107.0%;.The intra-day and inter-day RSDs were less than 9%. Conclusion: The method of HPLC in determining ATX was sensitive,specific and simple,the result shows that the two tablets are bioequivalent.