Rapid and simple determination of mycophenolic acid in human plasma by ion-pair RP-LC with fluorescence detection.
Mycophenolic acid (MPA) is an immunosuppressive drug given as the prodrug of mycophenolate mofetil (MMF). In order to investigate the pharmacokinetics of MPA, a simple, specific, sensitive and reliable method has been established for the quantitative determination of MPA in plasma from renal transplant recipients. The method involves a single-step protein precipitation procedure and a specific determination by ion-pair reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Separation was achieved on a C18 column (150 x 4.6 mm, 5 microm) with a mobile phase composed of borate buffer (pH 10.0; 50 mM)--acetonitrile--tetrabutylammonium bromide (200 mM) (75:25:1, v/v/v). The fluorescence detector was set at 310 (excitation) and 430 nm (emission). Following protein precipitation with ice-cold acetonitrile, clear supernatants (50 microl) were injected into the HPLC system. The retention time of MPA was approximately 4.5 min. The HPLC run time was 8 min. The assay was linear in concentration range 0.2-20.0 microg/ml for MPA in human plasma. Precision of the assay in the concentration range examined was from 0.89 to 3.21% for the intra-assay run and from 3.01 to 4.35% for the inter-assay run. A limit of detection was 0.05 microg/ml at a signal-to-noise ratio of 3. This validated method was then applied to the determination of MPA concentrations in renal transplant recipients after oral administration of 0.75 g of MMF.
Laboratory for Clinical Investigation, Osaka University Hospital, Suita, Osakafu, Japan.
H
H
Hosotsubo
S
S
Takahara
Y
Y
Kokado
S
S
Permpongkosol
J D
JD
Wang
T
T
Tanaka
K
K
Matsumiya
M
M
Kitamura
A
A
Okuyama
H
H
Sugimoto
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D007166:Immunosuppressive Agents / Q000097:blood*; D016030:Kidney Transplantation; D009173:Mycophenolic Acid / Q000097:blood*; D012680:Sensitivity and Specificity
D007166:Immunosuppressive Agents; D009173:Mycophenolic Acid