Luo Rongning, Li Zhangwan, Qian Guangsheng, Lu Jia, Fu Chunmei
Key Laboratory of Drug Targeting and Delivery System, West China School of Pharmacy, Sichuan University, Chengdu, PR China.
Yakugaku Zasshi. 2009 Dec;129(12):1545-9. doi: 10.1248/yakushi.129.1545.
A simple, sensitive, selective and reproducible reversed-phase HPLC method was developed for the determination of sophoricoside in rat plasma after intravenous administration. Naringin was successfully used as internal standard (IS) for calibration. The chromatographic separation was accomplished on a reversed-phase C(18) column using acetonitrile-methanol-0.08% phosphoric acid (8:29:63, v/v/v) as mobile phase with a flow rate of 1.0 ml/min, with UV detection at 260 nm. Plasma samples were injected into the HPLC system after precipitating protein directly by methanol. Good linearity was achieved in the range of 0.0240 approximately 48.0 microg/ml (R(2)=0.9989). The limit of detection (LOD) and limit of quantification (LOQ) of this method were 0.0075 microg/ml and 0.0240 microg/ml, respectively. The absolute recoveries of sophoricoside from plasma were 95.8%, 93.2%, 98.0% at concentrations of 0.0240, 1.92, 15.0 microg/ml. The intra-day and inter-day variabilities were 3.39%5.78% and 2.17%4.72%, respectively. The developed method was successfully applied to the pharmacokinetic study of sophoricoside after intravenous administration of 2.5, 10 and 20 mg/kg in rats.
建立了一种简单、灵敏、选择性好且可重现的反相高效液相色谱法，用于测定大鼠静脉给药后血浆中的槐角苷。柚皮苷成功用作内标物进行校准。色谱分离在反相C(18)柱上进行，以乙腈-甲醇-0.08%磷酸(8:29:63, v/v/v)为流动相，流速为1.0 ml/min，在260 nm处进行紫外检测。血浆样品经甲醇直接沉淀蛋白后注入高效液相色谱系统。在0.0240～48.0 μg/ml范围内线性良好(R(2)=0.9989)。该方法的检测限(LOD)和定量限(LOQ)分别为0.0075 μg/ml和0.0240 μg/ml。槐角苷在血浆中浓度为0.0240、1.92、15.0 μg/ml时的绝对回收率分别为95.8%、93.2%、98.0%。日内和日间变异系数分别为3.39%～5.78%和2.17%～4.72%。所建立的方法成功应用于大鼠静脉注射2.5、10和20 mg/kg槐角苷后的药代动力学研究。
