RP-HPLC测定黄芩素及其代谢产物黄芩苷大鼠血浆浓度

 目的建立用HPLC测定黄芩素/HP-β-CD包合物经大鼠尾静脉快速注射给药后,黄芩素及其代谢产物黄芩苷血浆浓度的方法。方法采用Agilent 1100 HPLC系统,Diamonsil^TMODS色谱柱,以甲醇和磷酸溶液组成的流动相梯度洗脱分离,以对羟基苯甲酸丙酯为内标,流速1.0 mL·min^-1,检测波长270 nm,柱温40℃,进样20μL检测。结果在本色谱条件下黄芩素、黄芩苷及其他代谢产物与杂峰分离良好;黄芩素和黄芩苷在0.05～15 mg·L^-1内呈线性关系;两者日间和日内精密度均低于11.6%;黄芩素和黄芩苷的最低检测浓度分别为0.03和0.05 mg·L^-1。结论该方法可在同一条件下同时检测黄芩素和黄芩苷,且简便、准确、可靠,可用于黄芩素/HP-β-CD包合物静注给药后,其主药黄芩素及其主要代谢产物黄芩苷的血药浓度测定。

Simultaneous Determination of Baicalein and Baicalin in Rat Plasma by RP-HPLC

OBJECTIVE To develop a RP-HPLC method with UV detection for the simultaneous determination of baicalein and its major metabolite(baicalin) after intravenous administration of baicalein/HP-β-CD complex. METHODS The Angilent 1100 HPLC system was used in the drug analysis.The samples were separated on a reversed-phase HPLC column(Diamonsil^TM ODS) with the mobile phase consisting of methanol and 0.2% phosphoric acid solution,using a linear gradient elution program.Propyl paraben was used as internal standard.The flow rate was set at 1 mL·min^-1 and the eluent was detected at 270 nm.The column temperature was 40 ℃ and 20 μL sample was injected into the HPLC column.RESULTS Both baicalein and baicalin had good linear relationship over the range of 0.05～15 mg·L^-1.The intra-day and inter-day RSD were less than 11.6%.The limits of quantification in plasma of baicalein and baicalin were 0.03 and 0.05 mg·L^-1,respectively.CONCLUSION The developed method is simple,accurate and reliable for determining baicalein and baicalin simultaneously in a single chromatographic run.