RP-HPLC内标法测定大鼠血清红景天苷浓度的研究

Determination of Salidroside in Rat Serum by RP-HPLC

目的RP-HPLC内标法建立大鼠血清中红景天苷浓度的测定方法。方法健康♂SD大鼠尾静脉注射红景天苷（60mg·kg^-1）后,分别于不同时间点眼眶静脉丛取血0.5mL,高氯酸沉淀血清蛋白,离心取上清液进样分析。色谱柱为SHIMADZU C18柱（4.6mm×250mm,5μm）,流动相为乙腈-水-三乙胺-冰醋酸（10∶90∶0.011∶0.016）,流速为1.5mL·min^-1,检测波长为277nm,柱温为50℃,进样量10μL,内标为咖啡因。结果红景天苷质量浓度在0.56-70mg·L^-1内线性关系良好（r=0.9975）,以信号/噪音=3∶1计算,红景天苷最低检测浓度为0.15mg·L^-1,日内、日间精密度分别≤4.5%,≤5.4%,血清样品的平均回收率为（99.27±4.38）%。结论建立的RP-HPLC内标法方法简便、快速、准确,适合于红景天苷的血药浓度测定及药动学研究。

OBJECTIVE To develop a HPLC assay for the determination of satidroside in rat serum. METHODS Each male rat received a dose of salidroside injection of 60 mg·kg^-1 by caudal vein injection. 0.5 mL venous blood samples from orbital vein were collected into polypropylene tubes after iv bolus administration.The serum was pretreated by precipitating with perchloric acid. A SHIMADZU C18 column was used. The mobile phase consisted of acetonitrile, water, triethylamine and glacial acetic acid （10 ： 90 ： 0.011 ： 0.016）.The flow rate was 1.5 mL·min^-1, and the UV detection was set at 277 nm.The column temperature was 50℃ and the sample size was 10 pL.Caffeine was used as the internal standard. RESULTS The linear calibration curve was obtained in the concentration range of 0.56-70 mg·L^-1 （r=0.997 5）. The Lower limit of quantitation （S/N=3） of the method was 0.15 mg·L^-1.The intra-day and inter-day RSD were within 4.5% and 5.4% respectively. And the range of the mean recovery in serum at concentrations of 5, 20 and 56 mg·L^-1 was （99.27±4.38） %. CONCLUSION The method is simple, rapid and suitable for the determination of the concentration of salidroside in rat serum and the study of pharmacokinetics.