柱前衍生化高效液相色谱法测定三七及其制剂中三七素含量

Analysis of dencichine by HPLC with pre-column derivatization

目的：建立三七药材及其制剂中三七素含量测定的方法。方法：以硼砂缓冲液（pH9．18）为提取溶剂，回流提取药材及制剂中的三七素；以氯甲酸芴甲酯（FMOC）作为衍生化试剂，对三七素提取液进行柱前衍生化，采用RP-HPLC，ODS柱分离，并通过紫外检测的方法进行测定。结果：在本实验条件下，三七素的线性范围为1．76-352mg·L^-1，最低检测限为60μg·L^-1。衍生化产物稳定，过量的衍生化试剂无干扰。平均加样回收率为95．3％，RSD为1．7％。结论：该方法操作简单，快速，灵敏度高，可用于三七药材及制剂中三七素的分析测定和三七素的药物代谢研究。

Objective：To establish a reversed-phase high performance liquid chromatorgraphy （RP-HPLC） method for detecting the dencichine in Panax notoginseng extracts and drug preparations. Method： Dencichine was extracted with the borate buffer （pH 9.18） and the clear supernatant was used for the derivatization. Pre-column derivatization was performed using 9-fluorenylmethyl chloroformate （FMOC） to form derivatives. The mobile phase consisted of methanol and 0.05 mol · L^-1 NaH2PO4 （48： 52） （pH adjusted to 7.4 with NaOH solution） in a flow rate of 1.0 mL · min^-1. The ultraviolet （UV） detection wavelength was set at 262 nm. Result：The linearity was demonstrated over a wide range of concentration from 1.76 mg · L^-1 to 352 mg · L^-1 for dencichine. The detection limit was determined to be 60 μg ·L^-1. The derivative was stable and the derivatization agent did not influence the measurement of dencichine. The average recovery rate was 95.3% and the relative standard derivation （RSD） was 1.7%. The method was used to determine dencichine in different P. notoginseng extracts and drug preparations. Conclusion： This method is simple, fast and sensitive, suitable for determining the dencichine in P. notoginseng extracts and drug preparations as well as for the study of the dencichine metabolism in vivo.