Abstract 4622: Determination of pyranocoumarin compounds decursin, decursinol angelate and decursinol simultaneously in mouse plasma and tumor tissue by liquid-liquid extraction and HPLC

Abstract

Abstract Pyranocoumarin compound decursin and its isomer decursinol angelate (DA) are the major chemical components of the root of Angelica gigas Nakai (AGN, Korean Angelica), a major traditional medicinal herb. Recently, AGN has been shown to exhibit anti-tumor activities by our collaborative team. Decursinol has been identified as a major metabolite of decursin and DA in vivo by us and others, but another group has published very controversial results. To better define the mechanisms of in vivo action for AGN, pharmacokinetic studies are essential. In this study, we set up a practical method for the concurrent analysis of decursin, DA and decursinol in mouse plasma and tumor tissues by liquid-liquid extraction and HPLC-UV. In brief, plasma or tumor homogenate (0.1-0.2 ml) spiked with 1μg chrysin as internal standard (IS) was extracted directly with ethyl acetate. After extraction, decursin, DA and decursinol were quantitated by HPLC with UV detection at 330 nm. The method was validated in terms of recovery, lower limit of quantitation and linear range. The extraction efficiency for spiked decursin/DA and decursinol at 1 µg/ml was 90.1±3.2 % and 90.3±1.8% in mouse plasma, respectively. The lower limit of quantitation (LOQ) was approximately 0.25 μg/mL for decursin/DA, and 0.2 μg/mL for decursinol in mouse plasma. The linear range of the method was 0.25 -25 µg/ml and 0.2-50 µg/ml in mouse plasma for decursin/DA and decursinol, respectively. In a pilot pharmacokinetic study, male C57BL/6 mice were given a single dose of 4.8 mg decursin/DA (∼240mg/kg) per mouse either by gavage or intraperitoneal injection in 0.2 mL vehicle of ethanol:PEG400:Tween80:5% glucose (3:6:1:20). Maximum plasma concentrations for decursin/DA and decursinol were found to be 34.2 and 324 µM respectively when D/DA was administered via intraperitoneal injection and to be 1.7 and 60.7 µM via gavage. These findings further confirmed that decursinol was one of the major metabolites of decursin/DA in vivo. In athymic nude mouse xenograft study, the mice were given intraperitoneal injection of decursin/DA at a dose of 1mg (∼40mg/kg) daily, 5 days per week, for 7 weeks. The decursin/DA and decursinol concentration was 1.5 and 10μg/g, respectively, in the tumor tissue. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4622. doi:10.1158/1538-7445.AM2011-4622