In the present study,we developed a rapid and specific reversed-phase high-performance liquid chromatographic(RP-HPLC)method for the quantification of p-hydroxyphenethyl anisate(HPA),one of the main bioactive constituents of the roots and rhizomes of Notopterygium incisum and N.franchetii,in rat plasma after an intravenous(20 mg/kg,i.v.)and an intragastrical(200 mg/kg,i.g.)administration to rats,respectively.The method involved a plasma clear-up step using liquid-liquid extraction by EtOAc,followed by RP-HPLC separation and detection.Separation of HPA was performed on an analytical DiamonsilTM ODS C18 column with the mobile phase of MeOH-H2 O at ratios of 75:25(v/v)for i.v.and 70:30(v/v)for i.g.administration.The flow-rate was 1.0 mL/min,and UV detection was performed at 256 nm.The calibration curves were linear over the ranges of 0.05-5.0μg/mL(r2=0.9984)for i.v.and 0.5-10.0μg/mL(r2=0.9995)for i.g.administration in rat plasma.The extraction recoveries were in the range of82.01%-87.97%.The intra-and inter-day precisions were between 1.71%and 3.99%,with accuracies ranging from 91.22%to110.5%.The absolute bioavailability of an orally administered HPA in rats was about 48.17%.The developed method was suitable for the determination and pharmacokinetic study of HPA in rat plasma.
Angelicae Pubescentis Radix (APR, Duhuo) is a commonly used traditional Chinese medicine and usually used for the treatments of rheumatic diseases. To assess the chemical composition of APR extract, a sensitive and reliable UPLC-Q?TOF-MS method was used for qualitative analysis. The separation was achieved on an Agilent SB-Ci8 column (1.8 pm, 2.1 mm><50 mm) with a gradient elution system consisting of acetonitrile and water containing 0.1% formic acid. An electrospray ionization (ESI) was used for mass spectrometer, and the data were collected in the positive ion mode, which was operated in a full-scan mode at m/z 100-800. A total of 49 compounds including 46 coumarins were identified according to the MS and MS/MS data. Among them, two were new compounds, and isoangenomalin, scoparone, 4-methyl-umbelliferyl acetate, suberenol,/raw5-dehydroosthol, and oroselone were first reported in APR.
Meiqi WanYoubo ZhanglYanfang YangXiaoyan LiuLingyun JiaXiuwei Yang
Angelicae Pubescentis Radix(APR), a widely used traditional Chinese medicine(TCM), is mainly used to treat rheumatism and headache diseases. Angelol B is one of the bioactive constituents of APR with significant anti-inflammatory activity. This paper is aimed to illustrate the metabolites of angelol B in vivo. To achieve this objective, a metabolomics approach based on a rapid and accurate UPLC-Q-TOF-MS method was used to detect the metabolites of Angelol B in rat. A gradient elution system(ACN and 0.1% formic acid water) equipped with an Agilent SB-C18 column(1.8 μm, 2.1 mm × 50 mm) to complete the separation. Scanning area at m/z 100-800 operated on an electrospray ionization(ESI). The data were collected in both positive and negative ion mode and analyzed by the Masslynx 4.1 and SIMCA 13.0 software. A total of 31 metabolites including 20 phase Ⅰ and 11 phase Ⅱ metabolites were identified. Their structure and fragmentation process were deduced based on the MS and MS/MS data. All of thirty-one metabolites are new compounds based on the search of SCI-Finder database.
WAN Mei-QiZHANG You-BoLIU Xiao-YanLI Ke-MingJIA Ling-YunYANG Xiu-Wei
