Measurement of daphnoretin in plasma of freely moving rat by liquid chromatography

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Title:	Measurement of daphnoretin in plasma of freely moving rat by liquid chromatography
Authors:	Lin, LC;Yang, KY;Chen, YF;Wang, SC;Tsai, TH
Contributors:	中醫學院中醫所 Natl Res Inst Chinese Med, Taipei 112, Taiwan;Natl Yang Ming Univ, Inst Tradit Med, Taipei 112, Taiwan;Natl Chung Cheng Univ, Dept Chem & Biochem, Chiayi 621, Taiwan;China Med Univ, Graid Inst Chinese Med Sci, Taichung, Taiwan
Date:	2005
Issue Date:	2010-09-20 13:44:11 (UTC+8)
Publisher:	ELSEVIER SCIENCE BV
Abstract:	Daphnoretin (7-hydroxyl-6-methoxy-3,7'-dicoumaryl ether), isolated from Wikstronemia indica C.A. Mey. (Thymelaceae), has been reported to induce rabbit platelet aggregation through protein kinase C activation and anticancer activity. In this study, we developed an automated blood sampling system coupled to a simple and sensitive HPLC system to determine plasma concentration of daphnoretin in rats. This method was applied to investigate the pharmacokinetics of daphnoretin in a freely moving rat. Separation of daphnoretin in the rat plasma was achieved using a reversed-phase C-18 column (250 mm x 4.6 mm, 5 μ m) with a mobile phase of methanol-10 mM NaH2PO4 (adjusted to pH 3.0 with H3PO4) (55:45, v/v), and the flow rate of 1.0 ml/min. The UV detector was set at 345 nm. The automated blood sampling system (DR-II has been applied for blood sampling in a conscious and freely moving rat. The blood samples were centrifuged at 3000 x g for 10 min and the plasma samples were then deproteinized by acetonitrile containing an internal standard (khellin 1 μ g/ml). After centrifugation (8000 x g for 10 min), the aliquot of supernatant was injected into the HPLC system for analysis. The concentration-response relationship from the present method indicated linearity over a concentration range of 0.05-1.00 and 1.00-100 μ g/ml. Intra- and inter-assay precision and accuracy of daphnoretin fell well within the predefined limits of acceptability (&LE; 15%). After daphnoretin (500 mg/kg) was given orally, the maximum concentration was 0.17 μ g/ml at the time of 5 min. The oral bioavailability was about 0.15 &COPY; 2004 Elsevier B.V. All rights reserved.
Relation:	JOURNAL OF CHROMATOGRAPHY A 1073(1月2日):285-289
Appears in Collections:	[Graduate Institute of Chinese Medical Science] Proceedings

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