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[摘要]
目的 建立一测多评法测定丹酚酸A原料药中特定杂质迷迭香酸、紫草酸、丹酚酸B和丹酚酸C。方法 采用Agilent Zorbax Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5 μm),流动相0.2%磷酸–乙腈,梯度洗脱,检测波长286 nm,体积流量1.0 mL/min,柱温25℃,样品室温度4℃,进样量20 μL。以丹酚酸A为内标,建立丹酚酸A原料药中特定杂质(紫草酸、迷迭香酸、丹酚酸B、丹酚酸C)的相对校正因子;分别采用一测多评法和外标法测定丹酚酸A原料药中特定杂质,并比较一测多评法计算值与外标法实测值的差异性。结果 丹酚酸A、丹酚酸B、丹酚酸C、迷迭香酸、紫草酸均在0.2~20 μg/mL与峰面积呈良好线性关系。在线性浓度范围内,迷迭香酸、紫草酸、丹酚酸B、丹酚酸C与丹酚酸A间的相对校正因子分别为1.52、2.09、2.33、1.06。一测多评法计算值与外标法实测值无显著性差异。结论 一测多评法可用于丹酚酸A原料中迷迭香酸、紫草酸、丹酚酸B、丹酚酸C的测定。
[Key word]
[Abstract]
Objective To establish quantitative analysis of multi-components with single marker (QAMS) for determination of specified impurities, such as rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C in salvianolic acid A crude drug. Methods HPLC method was carried out on Agilent Zorbax Eclipse XDB-C18 column(250 mm×4.6 mm, 5 μm). The mobile phase consisted of 0.2% phosphoric acid and acetonitrile solution with gradient elution. The detection wavelengths were 286 nm. The flow rate was 1.0 mL/min with injection volume of 20 μL. The column temperature and sample room temperature was set at 25℃ and 4℃, respectively. Salvianolic acid A was used as the internal reference substance to establish determination of specified impurities (rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C) in salvianolic acid A crude drug. The relative correlation factors (RCFs) of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were calculated and established. The contents of specified related substance were determined by the external standard method (ESM) and QAMS method, respectively. The QAMS method was validated through comparison of the results obtained by the two different methods. Results The linear ranges of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were all 0.2-20 μg/mL. The values of RCFs of rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid C were 1.52, 2.09, 2.33, and 1.06, respectively. The two methods had no significant difference in content results. Conclusion QAMS method can be used for the determination of specified impurities (lithospermic acid, rosmarinic acid, salvianolic acid B and salvianolic acid C) in salvianolic acid A crude drug.
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[基金项目]
北京市国家重大研发计划匹配项目(Z161100002616024)