HPLC-荧光检测法测定枸杞子和五子衍宗丸中东莨菪内酯的含量

Determination of Scopoletin in Lycii Fructus and Wuzi Yanzong Pill by HPLC-Fluorescence Detection

关键词:  高效液相色谱-荧光检测法  枸杞子  五子衍宗丸  东莨菪内酯

Key words:  HPLC-FLD  Lycii Fructus  Wuzi Yanzong pill  scopoletin

摘要:

目的 采用高效液相色谱-荧光检测法(HPLC-FLD)测定枸杞子和五子衍宗丸中东莨菪内酯的含量。方法 色谱条件：色谱柱为Agilent Extend-C₁₈柱(4.6 mm×250 mm，5 μm)；流动相为乙腈-0.1%磷酸(18∶82)；流速为1.0 mL?min^?1；柱温30 ℃；荧光检测器激发波长343 nm，发射波长458 nm。结果 东莨菪内酯在41.63~2 082 pg内线性关系良好(r=1)，回归方程Y=44 095X–44 660，在枸杞子药材中平均回收率为101.1%，RSD为2.2%(n=9)，在五子衍宗丸中平均回收率为102.6%，RSD为1.1%(n=9)。宁夏产的枸杞子东莨菪内酯含量高于其他产地。结论 该方法快速、简便、灵敏，专属性、重复性良好，有助于评价枸杞子的道地性以及初步区分样品的来源，适用于枸杞子药材和五子衍宗丸中君药枸杞子的质量控制与评价。

Abstract:

OBJECTIVE To develop an HPLC-fluorescence detection(HPLC-FLD) method for the determination of scopoletin in Lycii Fructus and Wuzi Yanzong pill. METHODS The chromatographic separation was performed on an Agilent Extend-C₁₈ column (4.6 mm×250 mm, 5 μm) with the mobile phase of acetonitrile-0.1% phosphoric acid (18∶82). The flow rate was 1.0 mL?min^-1at 30 ℃, and the detection was performed at wavelengths of 343 nm and 458 nm for excitation and emission, respectively. RESULTS The calibration curve was linear in the range of 41.63-2 082 pg (r=1), and the regression equation was Y=44 095X-44 660. The average recovery was 101.1% in Lycii Fructus, and 102.6% in Wuzi Yanzong Pill with the RSD of 2.2% and 1.1%, respectively. According to the assay result, the contents of scopoletin in Lycii Fructus collected from Ningxia province were higher than those from other origin. CONCLUSION The method is rapid, simple, sensitive and shows good specificity and reproducibility. It could be used to evaluate the genuineness of Lycii Fructus and control the quality of Lycii Fructus and Wuzi Yanzong pill.