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目的建立反相高效液相色谱法,同时测定人血浆中霉酚酸(mycophenolic acid,MPA)、酚化葡萄糖醛麦考酚酸(phenol glucuronide metabolite,MPAG)、酰基化MPAG(acyl-MPAG,AcMPAG)的浓度。方法用蛋白沉淀法对样品进行处理。固定相为Zorbax Eclipse XDB C18柱(4.6 mm×250 mm,5μm),流动相:甲醇:20 mmol-L-1 NaH2PO4(用20%磷酸调至pH 3.0)为55∶45,流速:1.2 mL-min-1,检测波长:304 nm,柱温:45℃。结果 MPA、MPAG、AcMPAG浓度在0.2~50μg-mL-1(r=0.999 7)、2.8~531μg-mL-1(r=0.999 9)、0.3~24μg-mL-1(r=0.999 4)内呈良好的线性关系。MPA及其代谢物的绝对回收率均大于80%,MPA、MPAG、AcMPAG的相对回收率分别为94.0%~101.4%,98.4%~101.9%和96.1%~104.2%。日内及日间RSD远低于15%。结论采用反相高效液相色谱法测定人血浆中MPA、MPAG、AcMPAG的浓度和进行药物代谢动力学研究,方法灵敏度高、重复性强。
Objective To establish a reversed-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous determination of mycophenolic acid (MPA), phenol glucuronide metabolite (MPAG), acyl-MPAG )concentration. Methods Samples were processed by protein precipitation. The stationary phase was a Zorbax Eclipse XDB C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of methanol: 20 mmol L -1 NaH 2 PO 4 (adjusted to pH 3.0 with 20% phosphoric acid) at 55:45 and a flow rate of 1.2 mL- min-1, detection wavelength: 304 nm, column temperature: 45 ℃. Results The concentrations of MPA, MPAG and AcMPAG were in the range of 0.2-50 μg-mL-1 (r = 0.999 7), 2.8-531 μg-mL-1 (r = 0.999 9) and 0.3-24 μg-mL-1 A good linear relationship. The absolute recoveries of MPA and its metabolites were all above 80%. The relative recoveries of MPA, MPAG and AcMPAG were 94.0% ~ 101.4%, 98.4% ~ 101.9% and 96.1% ~ 104.2%, respectively. RSD far below 15% during and after the day. Conclusion The concentration of MPA, MPAG and AcMPAG in human plasma were determined by RP-HPLC and the pharmacokinetics were studied. The method was highly sensitive and reproducible.