分离鉴定黄连花薹的差异性成分并测定其中4个成分的含量

Separation and analysis of characteristic components in Coptis chinensis flower moss and content determination of 4 components

目的采用薄层鉴别法与高效液相色谱法对比寻找出黄连药材与黄连花薹中含量最大的差异性成分,并对其进行鉴定;同时建立一测多评法测定黄连花薹中4个化学成分的含量。方法采用聚酰胺柱层析等分离纯化手段对黄连花薹与黄连药材差异性成分进行提取、分离、纯化,并采用液质联用以及核磁鉴定的方法对差异性成分进行确证;采用资生堂C18色谱柱(250 mm×4.6 mm,5μm),检测波长为340 nm,柱温为40℃,流速为1.0 mL·min^(-1),进样量为10μL,流动相为乙腈(A)-0.1%磷酸(每1000 mL加入20μL三乙胺)(B),梯度洗脱。以盐酸小檗碱为内标物,计算其他3个成分的相对校正因子,建立用一测多评法和常规外标法测定黄连花薹中4个化学成分的含量,并对两种方法的含量测定结果进行比较。结果分离鉴定出黄连花薹与黄连药材的差异性成分蒙花苷,同时在各自的线性范围(r>0.9999)内,芦丁、绿原酸、蒙花苷的相对校正因子分别为2.5704、2.4532、1.7700。6批黄连花薹一测多评法测定结果与外标法测定结果无显著差异,但大大提高了检测的效益,节约了检测成本。结论分离鉴定出黄连花薹与黄连药材的差异性成分蒙花苷,同时建立的一测多评法测定结果准确,可用于黄连花薹的定量分析及质量控制。

Objective To determine the largest content difference in components in Coptis chinensis and Coptis chinensis flower moss by thin layer identification method and high performance liquid chromatography,and to simultaneously determine 4 chemical components in Coptis chinensis by quantitative analysis of multi-components by single-marker(QAMS)method.Methods Different components were extracted and purified by polyamide column chromatography,and further confirmed by liquid mass spectrometry and nuclear magnetic identification.Shiseido C18 column(250 mm×4.6 mm,5μm)was used.The detection wavelength was 340 nm and the column temperature was 40℃.The flow rate was 1.0 mL·min^(-1),the sample size was 10μL,and the mobile phase was acetonitrile(A)-0.1%phosphoric acid(20μL triethylamine per 1000 mL)(B)gradient elution.Berberine hydrochloride was used as the internal standard,the relative correction factors of the other 3 components were calculated,and the content of 4 chemical components in Coptis chinensis was determined by QAMS and conventional external standard method.The results of the two methods were compared.Results The differential components of Coptis chinensis flower moss and Coptis chinensis were isolated and identified.At the same time,within the linear range(r>0.9999),ƒof rutin,chlorogenic acid,and linarin was 2.5704,2.4532,and 1.7700,respectively.Under different experimental conditions,the relative correction factor was relatively stable,without significant difference in the results of 6 batches of Coptis chinensis flower moss by QAMS and external standard method,but it greatly improved the detection efficiency and saved the detection cost.Conclusion The differential components of montanin are isolated and identified,and the QAMS method is accurate,feasible for the quantitative analysis and quality control of Coptis chinensis flower moss.