HPLC法测定大鼠血浆中川续断皂苷VI的浓度

Determination of Akebia Saponin D in Rat Plasma by HPLC

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 HPLC method for determination of akebia saponin D in rat plasma.METHODS：Plasma samples were determined by HPLC after precipitation with methanol.The separation was performed on a Diamonsil ODS column（250 mm × 4.6 mm,5 μm） with column temperature kept at room temperature with a mobile phase consisting of acetonitrile-methanol-water（32：17：51,v /v） at a flow rate of 1.0 mL·min-1.The ultraviolet detector was operated at 205 nm.6 Wistar rats were included in our study and which were givenakebia saponin D（55 mg·kg-1） intraperitoneally followed by determination of plasma concentration of akebia saponin D and calculation of pharmacokinetic parameters.RESULTS：The determination of Akebia saponin D was free of interference from endogenous substances in rat plasma.The linear range was 5.0 ～ 500.0 μg·mL-1（correlation coefficient r = 0.994 6） with a low quantization limit of 5.0 μg·mL-1.A mean extraction recovery of 85.0% was achieved（85.0%,83.6% and 86.6%,respectively at low,medium,and high concentrations）.Both intra-day and inter-day RSD values were below 7.8 %（n = 9）.Data processing carried out using non-compartmental model showed that the pharmacokinetic parameters were in line with pharmacokinetic tendency.CONCLUSION：This method is accurate and simple,and it can be used forconcentration measurement and pharmacokinetic studyof akebia saponin D in rat plasma.