A simple HPLC method for the quantification of mizoribine in human serum: pharmacokinetic applications

Abstract

A simple high-performance liquid chromatographic (HPLC) method was developed and validated for the quantification of mizoribine in human serum. After the addition of 70% perchloric acid and 3-methylxanthine (50 microg/mL, internal standard) to human serum, the samples were mixed and centrifuged at 12,000 rpm (1432 g) for 10 min. The supernatant was injected onto a C(18) column eluted with a mobile phase of 20 mm Na2HPO4 and methanol (93:7, v/v, pH 3) containing 0.04% octanesulfonic acid and detected utilizing an ultraviolet detector at 275 nm. The linear calibration curve was obtained in the concentration range of 0.1-4.0 microg/mL and the lower limit of quantification was 0.1 microg/mL. This method was validated with selectivity, linearity, precision and accuracy. In addition, the method was successfully applied to estimate the pharmacokinetic parameters of mizoribine in Korean subjects following an oral administration of 100 mg mizoribine (two Bredinine 50 mg tablets). The maximum serum concentration (C(max)) of 2.30 +/- 0.83 microg/mL was reached 2.27 +/- 0.66 h after an oral dose. The mean AUC(0-12 h) and the elimination half-life (t(1/2)) were 13.2 +/- 4.79 microg h/mL and 3.10 +/- 0.74 h, respectively.

Determination of Mizoribine in Human Plasma Using High-performance Liquid Chromatography: Application to a Pharmacokinetic Study in Chinese Renal Transplant Recipients

“…No organic reagent was used in the mobile phase, because organic phase, such as methanol and acetonitrile, would shorten the retention time of mizoribien and IS signifi cantly. In the method reported by Choi et al [ 19 ] , a mobile phase of 20 mM Na 2 HPO 4 and methanol (93:7, v/v, pH 3) containing 0.04 % octanesulfonic acid was used. Mizoribine was eluted early with the retention time of 2.38 min and slightly overlapped with a large chromatographic peak from endogenous compounds.…”

“…The ion-pair HPLC method developed by Erdmann et al [ 17 ] was validated with an IS, but the pretreatment for blood samples was complicated and time-consuming. Choi et al [ 19 ] Fig. 1a , also known as Bredinin ® , is an orally available immunosuppressive agent isolated from the soil fungus Eupenicillium brefeldianum [ 1 ] .…”