一测多评法测定小柴胡颗粒中7种成分

Study on determination of seven components in Xiaochaihu Granules by QAMS

目的建立一测多评法同时测定小柴胡颗粒中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、甘草酸、柴胡皂苷B2、柴胡皂苷B1的含量。方法建立高效液相色谱(HPLC)法检测小柴胡颗粒中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、甘草酸、柴胡皂苷B2、柴胡皂苷B1的含量,色谱柱为Waters XBridge C18柱(150 mm×4.6 mm,3.5μm),流动相为甲醇(A)-0.1%磷酸溶液(B),梯度洗脱,体积流量0.8 mL/min,检测波长254 nm,柱温35℃,进样量10μL,并进行专属性、线性关系、重复性、精密度、加样回收率等方法学验证;以黄芩苷为内参物,建立其与汉黄芩苷、黄芩素、汉黄芩素、甘草酸铵、柴胡皂苷B2和柴胡皂苷B1的相对校正因子,并进行含量测定。采用外标法测定15批小柴胡颗粒中的7个指标性成分对一测多评法的准确性进行验证。结果建立的HPLC法方法学验证结果符合要求;以黄芩苷为内参物,汉黄芩苷、黄芩素、汉黄芩素、甘草酸铵、柴胡皂苷B2、柴胡皂苷B1相对于黄芩苷的相对校正因子分别为0.78、0.52、0.57、1.80、0.82、1.08。一测多评法所测结果与外标法无显著性差异。结论该方法简单、可靠、耐用性良好,可用于小柴胡颗粒的质量控制。

Objective To establish a quantitative analysis of multi-components by single-marker(QAMS) method for the simultaneous determination of baicalin, wogonoside, baicalein, wogonin, glycyrrhizic acid, saikoside B2, saikoside B1 in Xiaochaihu Granules. Methods A HPLC method was established for the determination of baicalin, wogonin, baicalein, wogonin, glycyrrhizic acid, saikosaponin B2 and saikosaponin B1 in Xiaochaihu granules. The chromatographic column was waters XBridge C18 column(150 mm × 4.6 mm, 3.5 μm), The mobile phase was methanol(A)-0.1% phosphoric acid solution(B). The flow rate was 0.8 mL/min, the detection wavelength was 254 nm, the column temperature was 35 ℃, and the injection volume was 10 μL. The relative correction factors of wogonoside, baicalein, wogonin, ammonium glycyrrhetate, saikosaponin B2 and saikosaponin B1 were calculated by using baicalin as an internal reference substance in this study. In order to validate the accuracy, 15 batches of Xiaochaihu Granules were determined and compared by both QAMS and external standard method(ESM). Results The validation results met the requirements. Using baicalin as the internal reference substance, the relative correction factors of wogonoside,baicalein, wogonin, ammonium glycyrrhetate, saikosaponin B2 and saikosaponin B1 were 0.78, 0.52, 0.57, 1.80, 0.82, 1.08,respectively. There was no significant difference between QAMS and ESM. Conclusion The simple, reliable and robust QAMS can be used for the quality control of Xiaochaihu Granules.