Rapid and simultaneous determination of mycophenolic acid and its glucuronide conjugate in human plasma by ion-pair reversed-phase high-performance liquid chromatography using isocratic elution.
A high-performance liquid chromatographic method has been developed for the simultaneous determination of mycophenolic acid (MPA) and its glucuronide conjugate (MPAG) in human plasma. The method involves protein precipitation with acetonitrile, followed by ion-pair reversed-phase chromatography on C18 column, with a 40 mM tetrabutyl ammonium bromide (TBA)-acetonitrile (65:35, v/v) mobile phase. A 20-microl volume of clear supernatant was injected after centrifugation, and the eluent was monitored at 304 nm. No interference was found either with endogenous substances or with many concurrently used drugs, indicating a good selectivity for the procedure. Calibration curves were linear over a concentration range of 0.5-20.0 microg/ml for MPA and 5-200 microg/ml for MPAG. The accuracy of the method is good, that is, the relative error is below 5%. The intra- and inter-day reproducibility of the analytical method is adequate with relative statistical deviations of 6% or below. The limits of quantification for MPA and MPAG were lower than 0.5 and 5.0 microg/ml, respectively, using 50 microl of plasma. The method was used to determine the pharmacokinetic parameters of MPA and MPAG following oral administration in a patient with renal transplantation.
Hosotsubo
H
H
Laboratory for Clinical Investigation, Osaka University Hospital, Suita, Japan.
Takahara
S
S
Kokado
Y
Y
Permpongkosol
S
S
Wang
J D
JD
Tanaka
T
T
Matsumiya
K
K
Kitamura
M
M
Okuyama
A
A
Sugimoto
H
H
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005965:Glucuronates / Q000097:blood* / Q000493:pharmacokinetics; D020719:Glucuronides; D006801:Humans; D009173:Mycophenolic Acid / Q000031:analogs & derivatives / Q000097:blood* / Q000493:pharmacokinetics; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D005965:Glucuronates; D020719:Glucuronides; C113145:mycophenolic acid glucuronide; D009173:Mycophenolic Acid