高效液相色谱法测定黑种草子中黑种草苷含量

摘要

目的 建立黑种草子中黑种草苷的含量测定方法,为其开发利用提供依据.方法 采用高效液相色谱(HPLC)法,色谱柱为YMC-Pack ODS-A(250 mm×4.6 mm,5 μm);流动相为乙腈(A)-0.0175 mol·L-1乙酸水溶液(B),梯度洗脱(0~20 min,13%→18% A,>20~25 min,18%→13% A);流速1.0 mL·min-1;柱温30℃;检测波长290 nm.结果 黑种草苷在0.01~0.30 mg·mL-1范围内,峰面积与进样浓度呈良好的线性关系(R2＝ 0.9991);精密度、稳定性、重复性实验RSD均小于2%;平均回收率为96.66%,RSD为1.25%(n＝6).结论 该方法重复性好,准确性高,可用于黑种草子药材的质量控制.%Objective To establish a method for the determination of nigeglanoside in seeds of Nigella glandulifera. Methods The content of nigeglanoside was determined by HPLC.The separation was performed on a C18column ( YMC-Pack ODS-A,250 mm×4.6 mm,5 μm) with a gradient elution system of acetonitrile and 0.017 5 mol·L-1acetic acid solution at the flow rate of 1.0 mL·min-1.The detection wavelength was set at 290 nm,and column temperature was 30 ℃. Results The linear range of nigeglanoside was 0.01-0.30 mg·mL-1(R2＝0.9991).The RSDs of precision,stability and repeatability were all less than 2%.The average recovery was 96.66% (RSD＝1.25%,n＝6). Conclusion The method is accurate and reproducible. It is effective in controlling the quality of seeds of Nigella glandulifera .