HPLC法同时测定连翘中连翘苷和连翘酯苷A的含量

Simultaneous Determination of Phillyrin and Forsythoside A in Forsythiae Fructus by HPLC

目的建立HPLC法同时测定连翘中连翘苷和连翘酯苷A的含量。方法在色谱柱AgilentZobaxSBC-18（4．6mm×250mm，5μm）上进行分离，柱温为30℃，流动相：A相为乙腈；B相为0．2％甲酸水溶液，梯度洗脱：0～28min，15％A：85％B，28～33min，15％→20％A：85％→80％B．33～38min，20％A：80％B，38～50min，20％→25％A：80％→75％B，50～60min，25％A：75％B，流速：1．0ml·min，检测波长：278nm。结果样品中2种成分得到良好分离，连翘苷和连翘酯苷A分别在0．01975～0．05925mg·ml-1（r=0．9997），和0．0998～0．2994mg·ml-1（r=0．9996）范围内进样量与峰面积呈良好的线性关系，回收率分别为99．49％和99．18％，t／SD分别1．19％和1．04％，样品溶液12h内稳定。结论该方法操作简单，结果准确．专属性强．可用于连翘药材的质量控制。

Objective To establish an HPLC method for the simultaneous determination of phillyrin and for- sythoside A in forsythiae fructus. Methods The separation was performed on an Agilent reversed-phase Zorbax SB- C i8 （5 μm,4.6 mm × 250 mm）column. The temperature of the column was 30 ℃. The mobile phase was composed of acetonitrile and 0.2% formic acid water-solution in a gradient elution mode：0 -28 rain, 15% A：85% B ;28-33 rain, 15% →20% A ： 85% →80% B ; 33-38 min, 20% A ： 80% B ; 38-50 min, 20% →25 % A ： 80% ~75% B, 50-60 min, 25% A ：75% B. The flow rate was 1.0 ml ·min-1. The detection wavelength was set at 278 nm. Results Phill- yrin and forsythoside A could be completely separated. In the system of HPLC, the calibration curves of phillyrin and forsythoside A were linear in the range of 0. 01975 -0.05925 mg .ml-l（ r = 0. 9997, n = 5） and 0. 0998 - 0. 2994 mg ·ml-1 （ r = 0. 9996, n = 5 ）, respectively. The average recovery （ n = 6） was 99.49% and 99. 18% with RSD values at 1.19% and 1.04%. Sample solution was stable within 12 h. Conclusion This method is sim- ple, accurate and highly specific. It can be used for quality control of forsythiae fructus.