Simultaneous determination of mycophenolic acid and its glucuronide in human plasma by HPLC / 第二军医大学学报

ABSTRACT

Objective: To establish a method for determining mycophenolic acid(MPA) and its glucuronide(MPAG) inhuman plasma. Methods: A high performance liquid chromatographic assay with diode array detection was developed, and thedetector wavelength was set at 254nm. The plasma sample purification was limited to protein precipitation with acetonitrile.The sample was separated on Hypersil ODS2 column (200 mm ?4. 6mm, 5?m), the mobile phase consisted of solution A(25% acetonitrile and 75% 0.02 mol/L KH2PO4,pH 3.0)and solution B(70% acetonitrile and 30% 0.02 mol/L K2HPO4,pH6.5)with gradient elution. The flow rate was 1. 0 ml/min. Results: Calibration curves of MPA and MPAG were linear be-tween 1. 0-5. 0 ?g/ml (r=0.994 4,n=6) and (2.5-100) ?g/ml (r= 0.999 5,n=7),respectively. The detection limit of MPAand MPAG were 0. 5?g/ml, and 1.0 ?g/ml, respectively. The mean recoveries of high, medium and low concentrations ofMPA were (95. 75?2.3l)%, (104.10?1.91) % and (98.11?4.24)%, and MPAG were (97.37?1. 43)%, (101.10?5. 41)%and (105. 44?7.59)%. Conclusion: The present study provides a reliable quantitative method for pharmacokineticstudy and monitor of MPA and MPAG.