一测多评法测定清热止咳颗粒及中间体中7种黄酮类成分的含量

Determination of seven flavonoids in Qingre Zhike Granules and intermediate product with quantitative analysis of multi-components by single marker

建立同时测定清热止咳颗粒及清热止咳颗粒中间体中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷和汉黄芩苷7种黄酮类成分的一测多评方法(QAMS)。采用高效液相色谱法,Thermo BETASIL C18色谱柱(4. 6 mm×250 mm,5μm),流动相乙腈-0. 1%甲酸水溶液(梯度洗脱),流速0. 8 m L·min-1,检测波长280 nm,柱温30℃。以新橙皮苷为内参物,建立该成分与芸香柚皮苷、柚皮苷、橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷和汉黄芩苷的相对校正因子,采用该相对校正因子计算其他6种成分的质量分数,同时与采用外标法测定的质量分数进行比较,验证QAMS的可行性和准确性。新橙皮苷与芸香柚皮苷、柚皮苷、橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷和汉黄芩苷的相对校正因子分别为1. 150,1. 109,1. 110,0. 658,0. 723和0. 572,在不同实验条件下相对校正因子重现性良好,7种成分的计算值与实测值无显著性差异。以新橙皮苷为内参物,同时测定芸香柚皮苷、柚皮苷、橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷和汉黄芩苷的QAMS准确且可行,可用于清热止咳颗粒及中间体的质量控制。

This study is to determine seven flavonoids(narirutin,naringin,hesperidin,neohesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside)in Qingre Zhike Granules and intermediate products by quantitative analysis of multi-components with a single marker(QAMS).High performance liquid chromatography(HPLC)was performed on Thermo BETASIL C18 chromatographic column(4.6 mm×250 mm,5μm)with acetonitrile-0.1%formic acid solution as mobile phase for gradient elution at a flow rate of 0.8 m L·min-1.The detection wavelength was set at 280 nm and the column temperature was 30℃.Six relative correction factors(RCFs)of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside were established in the HPLC method with neohesperidin as the internal standard,and then these RCFs were used to calculate the mass fractions of the six components.Such mass fractions were compared with the mass fraction determined by the external standard method(ESM)at the same time to verify the feasibility and accuracy of QAMS method.Within the linear range,the RCFs of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside were 1.150,1.109,1.110,0.658,0.723 and 0.572,respectively,with good repeatability in different experimental conditions.There was no significant difference between the QAMS method and ESM method.The QAMS method with neohesperidin as the internal standard is feasible and accurate for the determination of narirutin,naringin,hesperidin,baicalin,oroxylin A-7-glucoronide and wogonoside,and it can be used for quality control of Qingre Zhike Granules and intermediate products.