HPLC含量测定和DNA分子标记技术对不同产地金银花的鉴定

目的:通过指标成分含量测定及DNA分子标记技术对不同产地金银花样品进行鉴定,探讨DNA分子鉴定技术在中草药鉴别领域的应用前景。方法:以绿原酸、木犀草苷、灰毡毛忍冬皂苷乙为指标成分,利用HPLC测定指标成分含量。提取金银花、山银花对照药材及河北和山东产地金银花样品DNA,检测DNA浓度和纯度,采用基于聚合酶链式反应(PCR)方法的DNA分子标记技术鉴别样品。结果:河北和山东产地金银花中绿原酸质量分数3.1%~3.3%,河北、山东产地金银花中木犀草苷质量分数分别为0.049%~0.050%,0.069%~0.078%,2个产地样品中均未检出灰毡毛忍冬皂苷乙成分。采用基于PCR方法鉴定出河北和山东产地各3批样品均为金银花。结论:DNA分子鉴定技术作为含量测定常规方法的补充,具有准确可靠、重复性高等优点,可用于金银花药材的鉴定。

Identification of Lonicerae Japonicae Flos from Different Areas by HPLC Assay and DNA Molecular Markers

Objective: To identify Lonicerae Japonicae Flos from different areas by determination of index ingredients and DNA molecular markers. Method: High performance liquid chromatography method was used for determination of chlorogenic acid,galuteolin and macranthoidin B.Genomic DNA was extracted from control medicinal material of Lonicerae Japonicae Flos and Lonicerae Flos,as well as samples of Lonicerae Japonicae Flos from Hebei and Shandong,samples were identified by allele-specific diagnostic polymerase chain reaction (PCR) based on sequences of internal transcribed spacer. Result: The content of chlorogenic acid in samples from Heibei and Shandong was 3.1%-3.3%.The content of galuteolin in samples from Hebei and Shandong were 0.049%-0.050% and 0.069%-0.078%,respectively.Macranthoidin B was not detected in samples from both areas.Samples from both areas were identified as Lonicerae Japonicae Flos by diagnostic PCR. Conclusion: As a supplement to conventional methods,for identification of Lonicerae Japonicae Flos,DNA molecular technology has a great advantage in accuracy and good reliability.