High-performance liquid chromatography method for the determination of mycophenolic acid and its acyl and phenol glucuronide metabolites in human plasma.
Measuring the concentration of the pharmacologically active metabolite of mycophenolic acid (MPA), acyl-MPAG (AcMPAG), in addition to the pharmacologically inactive phenol glucuronide metabolite (MPAG) may prove useful in the therapeutic drug monitoring of MPA. A simple high-performance liquid chromatography method with ultraviolet detection (HPLC-UV) was established for simultaneous determination of MPA, AcMPAG, and MPAG in human plasma. The method utilizes 2 internal standards (IS), phenolphthalein glucuronic acid (PGA) for MPAG and a carboxy butoxy derivative of MPA (MPAC) for AcMPAG and MPA. The method consists of solid-phase extraction of the analytes followed by analysis over a Zorbax Rx C8 column (150 x 4.6 mm, 5 mum) at 254 nm. The analytes were separated with a gradient mixture of methanol and 0.1% phosphoric acid over a run time of 14 minutes at a flow rate of 1 mL/min. The assay was linear in the concentration range from 0.2 to 50 mg/L for MPA, 0.5 to 25 mg/L for AcMPAG, and 2 to 500 mg/L for MPAG. The mean +/- SD interday accuracy and %CV for MPA were 100.3 +/- 5.7 and 5.7%, for AcMPAG, 102.6 +/- 5.7 and 5.6%, and for MPAG 100.5 +/- 5.3 and 5.3%, respectively. The average +/- SD of MPA, MPAG, and AcMPAG maximum concentrations (Cmax) in 23 kidney transplant recipients on 500 or 1000 mg twice daily mycophenolate mofetil were 11.77 +/- 9.43, 88.15 +/- 46.4, and 3.01 +/- 1.73 mg/L, respectively, and the predose trough (Cmin morning) concentrations were 2.24 +/- 3.11, 55.44 +/- 29.55, and 1.42 +/- 0.74 mg/L, respectively. The method described is robust, sensitive, reproducible, and will be useful in therapeutic drug monitoring or pharmacokinetic studies of MPA.
Patel
Chirag G
CG
Clinical Pharmacokinetics Research Laboratory, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, USA.
Akhlaghi
Fatemeh
F
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005965:Glucuronates / Q000097:blood*; D006801:Humans; D016030:Kidney Transplantation; D009173:Mycophenolic Acid / Q000097:blood* / Q000378:metabolism
D005965:Glucuronates; D009173:Mycophenolic Acid