Pharmacokinetic study of free mangiferin in rats by microdialysis coupled with microbore high-performance liquid chromatography and tandem mass spectrometry

Abstract

Mangiferin (2-β- d-glucopyranosyl-1,3,6,7-tetrahydroxyxanthen-9-one) has been isolated from the herbal root of Anemarrhena asphodeloides Bung showing antioxidative, antiviral, and anticancer effect. An in vivo microdialysis sampling method coupled to microbore high-performance liquid chromatography (HPLC) was employed for continuous monitoring of free mangiferin in rat blood. Microdialysis probes were inserted into the jugular vein/right atrium and brain striatum of Sprague–Dawley rats, and mangiferin at doses of 10, 30 or 100 mg/kg were then administered via the femoral vein. Dialysates were collected every 10 min and injected directly into a microbore HPLC system. Mangiferin was separated by a reversed-phase C 18 microbore column (150×1 mm) from dialysate within 10 min. The mobile phase consisted of acetonitrile–0.05% phosphoric acid–tetrahydrofuran (10:75:15, v/v/v) with a flow-rate of 0.05 ml/min. The wavelength of the UV detector was set at 257 nm. The limit of quantification for mangiferin was 0.05 μg/ml and in vivo recovery of mangiferin at concentrations of 1, 5 and 10 μg/ml was in range of 37.7–39.8%. The results indicate that the pharmacokinetics of mangiferin at doses of 10–30 mg/kg reveals a linear relation, while doses of 30–100 mg/kg show a nonlinear pharmacokinetic phenomenon. Mangiferin was undetectable in brain dialysate. The proposed method provides a technique for rapid and sensitive analysis of free mangiferin in rat blood and further application in pharmacokinetic study. Furthermore, the metabolites of mangiferin in the rat bile were confirmed by LC electrospray ionization (ESI) tandem mass spectrometry (MS–MS).