金银花提取物的质量评价研究

Study on Quality Evaluation of Lonicerae japonicae Thunb. Extracts

【目的】采用特征图谱、绿原酸与木犀草苷含量测定、山银花检查等综合方法,评价市场金银花提取物的质量。【方法】高效液相色谱(HPLC)特征图谱及绿原酸含量测定以Kromasil 100-C18为色谱柱,流动相为乙腈—0.4%磷酸溶液(梯度洗脱),检测波长为327 nm,柱温为35℃,流速为1.0 m L·min^(-1)。木犀草苷HPLC含量测定以ZOBAX SB-Phenyl为色谱柱,流动相为乙腈—0.5%(体积分数)冰乙酸(梯度洗脱),检测波长为350 nm,柱温为25℃,流速为0.9 m L·min^(-1)。山银花HPLCELSD检查方法,以Kromasil 100-C18为色谱柱,流动相为乙腈—0.4%(体积分数)冰乙酸(梯度洗脱),流速为1.0 m L·min^(-1)。【结果】13批市售金银花提取物特征图谱均检出6个酚酸类特征峰,分别为新绿原酸、绿原酸、隐绿原酸、3,4-O-二咖啡酰基奎宁酸、3,5-O-二咖啡酰基奎宁酸、4,5-O-二咖啡酰基奎宁酸。金银花与山银花药材均检出6个酚酸类特征峰,但药材与提取物相对峰面积差异较大,提示提取物制备过程中发生酚酸类成分异构体的转化。13批样品中有6批样品检出山银花的灰毡毛忍冬皂苷乙特征峰,提示这些样品为山银花提取物伪品或掺假山银花提取物。样品中绿原酸与木犀草苷含量分别为1.56~45.61 mg·g^(-1)、0.038~0.734 mg·g^(-1),山银花提取物伪品或掺伪品的绿原酸含量相对较高,而木犀草苷含量明显较低,不同厂家的正品金银花提取物绿原酸含量差异也较大,提示可能与金银花原料质量、制备工艺方法及辅料加入量的不统一有关。【结论】市场上金银花提取物质量差异很大,山银花假冒金银花或掺伪金银花现象严重,应加强原料质量控制、确定稳定的制备工艺,以保障提取物质量的稳定性。

Objective To evaluate the quality of Lonicerae japonicae Thunb. extracts in China market by combinedmethods of characteristic spectrum analysis, content determination of cholorogenic acid and cynaroside, andidentification of Lonicera confusa DC.. Methods High performance liquid chromatography（HPLC）for characteristicspectrum and content determination of cholorogenic acid were performed on Kromasil 100-C18 column,the mobilephase was acetonitrile-0.4% phosphoric acid aqueous solution with gradient elution,the detection wavelength was327 nm,the column temperature was 35 ℃,and the flow rate was 1.0 m L·min^-1. HPLC for content determinationof cynaroside was performed on Zorbax SB-phenyl column, the mobile phase was acetonitrile-0.5%（volumefraction）glacial acetic acid aqueous solution with gradient elution, the detection wavelength was 350 nm, thecolumn temperature was 25 ℃,and the flow rate was 0.9 m L·min^-1. HPLC-evaporative light scattering detection（ELSD）for examination of Lonicera confusa DC. was performed on Kromasil 100-C18 column,the mobile phasewas acetonitrile-0.4%（volume fraction）glacial acetic acid aqueous solution with gradient elution,and the flowrate was 1.0 m L · min-1. Results Six characteristic peaks of phenolic acids were presented in the characteristicspectrum of 13 batches of Lonicerae japonicae Thunb. extracts,and they were neochlorogenic acid,chlorogenicacid, cryptochlorogenic acid, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, and 4,5-O-dicaffeoylquinic acid, respectively. The 6 characteristic peaks of phenolic acids were identified from both Lonicerae japonicae Thunb. and Lonicera confusa DC., but there were great differences in relative peak areasbetween original medical herbs and their extracts,showing that the transformation of isomers of phenolic acidsoccurred during the preparation for the extracts. In 13 batches of samples,characteristic peaks of macranthoidin Bof Lonicera confusa DC. had been identified from 6 batches,showing that Lonicera confusa DC. extracts were usedas the counterfeits or adulterants in these samples. The contents of cholorogenic acid and cynaroside in the sampleswere in the range of 1.56-45.61 mg·g^-1,0.038-0.734 mg·g^-1,respectively. But the contents of chlorogenic acidfrom the counterfeits or adulterants of Lonicera confusa DC. extracts were relatively higher, and the cynarosidecontent was much lower. The differences in chlorogenic acid content from original Lonicerae japonicae Thunb.extracts prepared by different manufacturers were obvious, which probably resulted from the improper qualitycontrol of Lonicerae japonicae Thunb. medicinal material, preparation process, and adjuvant contents.Conclusion The difference of quality of Lonicerae japonicae Thunb. extracts in China market varies greatly,andthe phenomenon of Lonicera confusa DC. being used as the counterfeit or adulterant for Lonicerae japonicae Thunb.is serious. Therefore,its quality control should be strengthened,and stable preparation process should be appliedto guarantee the stability of the quality of extracts.