在线固相萃取-HPLC法测定比格尔犬血浆中丹参素的含量

Determination of danshensu concentration in beagle dog plasma by on-line solid phase extraction-HPLC method

目的：建立测定比格尔犬血浆中丹参素含量的在线固相萃取-HPLC法。方法：血浆样品经蛋白质沉淀,采用Lichrospher C18为富集柱,以乙腈-10 mmol/L NaH2 PO4（5∶95）为富集流动相,流速2 ml/min;选择Ultimate XB-C18（50 mm×4.6 mm,5μm）为分析柱,以乙腈-10 mmol/L NaH2 PO4（11∶89）为分析流动相,流速为0.8 ml/min,检测波长285 nm。结果：丹参素在20-2 000 ng/ml浓度范围内线性关系良好,最低定量限为20.0 ng/ml,日内和日间RSD均〈10%,平均绝对回收率〉80%。结论：本方法简便、快速、灵敏、可靠,适用于比格尔犬血浆中丹参素含量的测定。

Objective：To establish an on-line solid phase extraction（SPE）-HPLC method for the determination of plasma concentration of danshensu in beagle dog.Methods： The plasma samples were treated by protein precipitation.A Lichrospher C18（37 mm×4.6 mm,25 μm） column was used as trap column,with the loading solvents of acetonitrile-10 mmol/L NaH2PO4（5∶95） at a flow rate of 2 ml/min.An Ultimate XB-C18（50 mm×4.6 mm,5 μm） column was used as analytical column,with the mobile phase of acetonitrile-10 mmol/L NaH2PO4（11∶89） at a flow rate of 0.8 ml/min.The detection wavelength was 285 nm.Results： The calibration curve was linear in the concentration range of 20-2 000 ng/ml.The low limit of quantification was 20 ng/ml.The inter-and intra-day RSDs were less than 10%.The average recovery rate was more than 80%.Conclusion： This method is simple,rapid,sensitive,reliable and suitable for the determination of plasma concentration of danshensu in beagle dog.