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目的:建立了高效液相色谱测定大鼠血浆中川续断皂苷VI浓度的方法。方法:血浆样品经甲醇沉淀蛋白后,采用HPLC法测定。色谱柱为Diamonsil C18柱(250mm×4.6mm,5μm);流动相为乙腈-甲醇-水(32:17:51,v/v);检测波长205nm;柱温为室温;流速1.0mL·min-1。动物实验使用Wistar大鼠6只,采用腹腔注射给药(55mg·kg-1),进行血药浓度测定和药动学参数计算。结果:大鼠血浆内源物质对药物测定无干扰,在血药浓度5.0～500.0μg·mL-1范围内,浓度对峰面积有良好的线性关系(r=0.9946),定量下限为5.0μg·mL-1;低、中、高3种浓度样品的回收率分别为85.0%、83.6%和86.6%,平均为85.0%;日内、日间精密度RSD均小于7.8%(n=9)。采用非房室模型分析方法进行数据处理,所得参数符合药物动力学变化趋势。结论:该法准确、简便,可用于川续断皂苷VI在大鼠血浆中的浓度测定和药物动力学研究。
OBJECTIVE: To establish a method for the determination of Dipsacus saponin VI in rat plasma by high performance liquid chromatography. Methods: Plasma samples were precipitated with methanol and determined by HPLC. The column was Diamonsil C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-methanol-water (32:17:51, v / v) 1. Animal experiments using 6 Wistar rats, intraperitoneal injection (55mg · kg-1), plasma concentration determination and pharmacokinetic parameters. Results: The rat plasma endogenous substances had no interference with the drug determination. There was a good linear relationship (r = 0.9946) between the concentration and peak area in the concentration range of 5.0-500.0μg · mL-1, the lower limit of quantification was 5.0μg · mL-1. The recoveries of low, middle and high concentration samples were 85.0%, 83.6% and 86.6% respectively, with an average of 85.0%. The intra-day and inter-day RSDs were less than 7.8% (n = 9). Data were processed by non-compartmental model analysis and the parameters were in accordance with the trend of pharmacokinetics. Conclusion: This method is accurate and simple and can be used to determine the concentration and pharmacokinetics of Dipsacus saponin VI in rat plasma.