超高效液相色谱法测定青娥丸中松脂醇二葡萄糖苷的含量

Determination of pinoresinol diglucoside in Qing’e Pills by ultra performance liquid chromatography

建立了超高效液相色谱(UPLC)测定青娥丸中主要活性成分松脂醇二葡萄糖苷(pinoresinol diglucoside,PDG)含量的方法。样品经索氏提取后,提取物再用Waters Oasis HLB SPE固相萃取小柱进行前处理以消除杂质对PDG色谱峰的干扰。色谱条件:采用Waters Acquity C18BEH UPLC柱(100mm×1.0mm,1.7μm)分离,以乙腈-水(使用磷酸调pH值至4.0)(9∶91,v/v)为流动相,流速为0.1mL/min,检测波长为227nm,柱温为25℃,进样量为0.5μL。在上述条件下,松脂醇二葡萄糖苷在1.40～506.00mg/L范围内呈现良好的线性关系,相关系数r=1;低、中、高3个添加水平下的平均回收率分别为100.51%、102.37%、100.10%(n=9)。该方法准确、灵敏、重现性好,可用于青娥丸的质量控制。

A new ultra performance liquid chromatographic （ UPLC） method was established for the de-termination of pinoresinol diglucoside （ PDG） in Qing’e Pills. After extracted by the Soxhlet’s method, the methanol extracts of the samples were passed through a Waters Oasis HLB SPE column to achieve good chromatographic performance. The separation was performed on a Waters Acquity C18 BEH column （ 100 mm ×1. 0 mm,1. 7 μm） with acetonitrile-water （ the pH adjusted to 4. 0 with phosphoric acid） （ 9 ∶ 91,v/v） as the mobile phase at a flow rate of 0. 1 mL/min. The detection wavelength was set at 227 nm,the column temperature was 25 ℃ and the injection volume was 0. 5 μL. Under the optimized condi-tions,there was good linear relationship between the mass concentration and the peak area of PDG in the range of 1. 40-506. 00 mg/L with the correlation coefficient of 1. The average recoveries of PDG at three levels ranged from 100. 10% to 102. 37%. The method is accurate,sensitive,highly reproducible and suitable for the quality control of Qing’e Pills.