HPLC法同时测定人尿液中霉酚酸及其代谢物葡糖苷酸结合物的浓度

Simultaneous determination of mycophenolic acid and its glucuronide metabolite in human urine by HPLC

目的用HPLC测定人尿液中霉酚酸(MPA)及其代谢物葡糖苷酸结合物(MPAG)的浓度。方法分析柱为Zorbax Eclipse XDB-C8(150 mm×4．6mm,5μm);流动相为甲醇-0．1％三氟乙酸溶液(55:45,V／V);尿稀释5倍后直接进样。MPAG采用紫外检测,检测波长为250nm;MPA采用柱后添加0．2 mol·L-1 NaOH溶液进行荧光检测,荧光激发波长325 nm,发射波长435 nm。结果MPA和MPAG的线性范围分别为0．1-50 mg·L-1和10-500 mg·L-1,MPA、MPAG日内和日间RSD均<10％。结论此法能简便、灵敏、准确地测定人体尿液中MPA与MPAG的浓度,可用于临床药动学研究和治疗药物浓度监测。

AIM To establish a method for determination of mycophenolic acid （MPA）and MPA-glucuronide （MPAG） in human urine by HPLC. METHODS The separation was carried out using a Zorbax Eclipse XDB-Cs analytical column with the isocratic elution system of methanol and water containing 0.1% trifluoroacetic acid（55：45, V/V）. The sample preparation procedures involved 5-fold dilution for urine. MPAG was determined by UV detection set at 250 nm while MPA was quantified by fluorescence detection after post-column addition of 0.2 mol·L^- 1 sodium hydroxide solution.The excitation and emission wavelengths were set at 325 nm and 435 nm,respectively. RESULTS Linearity was observed from 0.1 to 50 mg·L^-1 of MPA and 10 to 500 mg·L^-1 MPAG in urine. The intra-day variance and inter-day variance were 〈 10%. CONCLUSION The method reported is found to be rapid and simple for quantitation of MPA and MPAG,thus can be applied for clinical pharmacokinetic studies and therapeutic drug monitoring.