高效液相色谱法测定血浆6-巯基嘌呤质量浓度及其药动学

Determination of 6-mercaptopurine in human plasma by HPLC and study on its pharmacokinetics

目的：建立血浆中硫唑嘌呤的活性代谢物6-巯基嘌呤质量浓度测定的高效液相色谱分析方法，并研究了6-巯基嘌呤人体药动学。方法：6-巯基嘌呤血浆样品经乙腈沉淀蛋白并行萃取，进一步将有机层吹干富集药品。Hypersil C18柱（4．6mm×150mm，5μm），流动相为乙腈一水（含0．25％醋酸）（3：97），流速1．0mL·min^-1，柱温30℃，检测波长323nm。结果：6-巯基嘌呤在2～200μg·L^-1范围内线性关系良好（r=0．9997），血浆最低检测限度为2μg·L^-1。高、中、低质量浓度（200，50，5μg·L^-1，）的日内及日间RSD均小于8．52％，平均方法回收率分别为98．1％，101．6％和100．8％，平均提取回收率分别为90．2％，94．9％和91）．90％。人体药动学研究表明，口服硫唑嘌呤后6-巯基嘌呤的药一时曲线符合一级吸收的一房室模型。结论：此法灵敏、专一、准确、精密、简便，适用于药动学研究及治疗药物质量浓度监测。

OBJECTIVE To establish an HPLC method for the determination of 6-mercaptopurine in human plasma,which is the active metabolite of azathioprine, and to study its pharmacokinetics in healthy volunteers. METHODS 6-Mercaptopurine was extracted from human plasma with acetonitrile after deproteinized,and then detected by Hypersil C18 column（4. 6 mm× 150mam, 5μm） with UV detector at 323 nm. The mobile phase consisted of acetonitrile-water（0. 25% acetic acid）（3：97） with a flow rate of 1.0 mLo min^-1. RESULTS The calibration curve was linear over the range of 2-200μg·L^-1 （ r = 0. 999 7） and the limit of quantitation was 2μg·L analytical recoveries were 98.1% The intra- and inter-assay coefficients of variation were less than 8. 52% at all level. The mean 1%, 101.6 % and 100. 8 %, respectively. The mean extraction recoveries were 90.2 %, 94. 9 % and 90.9%,respectively. The results showed the concentration-time curves of 6-MP were fitted to one-compartment model with first order absorption. CONCLUSION This method is sensitive, specific, accurate, precise and rapid, and is suitable for the pharmacokinetic study of 6-MP and therapeutic drug monitoring.