Research On Genuineness And Quality Evaluation Of Forsythia Suspensa Produced From Shanxi

Objective:The forsythia fruit from Shanxi used as the research object compared with forsythia from the Shaanxi, Hebei, Henan origin. And the study of genetic information, trace elements,active component and biological effects in forsythia was carried out, Firstly,RAPD technique has been used to construct DNA fingerprints and analyze genetic diversity; Secondly, methods of determination of trace elements in Forsythia suspensa were established, using for the analysis of trace elements in different origin of Forsythia suspensa.The contents of volatile oil,lignans,phenethyl alcohol glycosides,flavonoids and other components from Forsythia suspensa in different origins were analyzed by FHLC and GC.In addition,we investigated the anti-biotic of volatile oil and water decoction from different areas Forsythia suspensa and anti-influenza A virus (H1N1) activity in vitro of different areas Forsythia suspensa. Finally,based on integrated use of DNA, GC,HPLC technology and the QAMS,we established quality evaluation system of Shanxi forsythia herbs.Methods:1.DNA is extracted from fresh fruit of Forsythia suspensa,The primers with good polymorphism were screened from 45 RAPD primers by RAPD-PCR technology,using for DAN Fingerprinting Construction and Cluster analysis,NTSYS-pc2.1 software is applied to estimate GS between materials by UPGMA analysis.2.Volatile oil was extracted by Steam distillation method, major components(a-Pinene,Camphene,β-Pinene,Myrcene,P-cymene,Cinene,α-Terpineol) of Forsythia suspensa in different origins were analyzed by GC,Chemical components of Forsythia were extracted by reflux extraction method and major components(Pinoresinol-β-D-glucoside,Forsythiaside A,Forsythiaside B,Rutin,Forsythin,Phillygenin) of Forsythia suspensa in different origins were analyzed by HPLC.3.K,Mg,Ca, Na, Mn, Fe, Cu, Zn in Forsythia suspensa were directly determined by AAS with digesting agent HN03-HC104(4:1).4.Using the antibacterial circle diameter as the evaluation index,antibacterial activity of volatile oil and water decoction was study by the K-B drug sensitive slips method;Drugs were used MDCK cell toxicity and anti influenza A virus activity experiment,SI is used as the evaluation index,Forsythia suspensa of different areas study in vitro anti influenza A virus (H1N1) activity.5.Cinene was used as the internal reference,the calibration factor of other five constituents(α-Pinene,Camphene,β-Pinene,Myrcene,a-Terpineol)was calculated by GC,comparing difference between the caculated values of QAMS and the measured values and of ESM, The relative error proves reliability of determination of six components content by QAMS;Forsythinwas used as the internal reference,we calculated respectively the calibration factor of other five constituents((Pinoresinol-β-D-glucoside,Forsythiaside A,ForsythiasideB,Rutin,Phillygenin)was calculated by HPLC, comparing difference between the caculated values of QAMS and the measured values of ESM, The relative error proves reliability of determination of six components content by QAMS.Results:1.11 RAPD primers with good polymorphism were screened from 45 RAPD primers,PCR amplified 80 bands with the average7.27 bands for each primer,Of which 67 PPB (ratio of 83.8%),which was an effective molecular marker.The result of cluster analysis showed that Forsythia herbs can be divided into a single class of Shanxi and Hebei,Henan, Shaanxi clustered in another,when GS is 0.672.2.the result showed there was significant differences in the contents of the major components of volatile oil from Forsythia suspensa in different origins,and main ingredients of forsythia volatile oil were higher than Henan, Hebei, Shaanxi;The contents of major components of Forsythia suspensa in different origins were also significant difference,and(Pinoresinol-β-D-glucoside,ForsythiasideB,ForsythiasideA,Forsythin,Phillyg enin) main components of forsythia volatile oil were higher than Henan, Hebei, Shaanxi.3. AAS method determinated the trace elements in Forsythia suspensa, the method of AAS has well accuracy and reproducibility,and these trace elements from different origins are difference.4.The anti-biotic of volatile oil and water decoction and anti-influenza A virus (H1N1) activity in vitro from Shanxi areas Forsythia suspensa were better than Henan, Hebei, Shaanxi.5. The result of contents of the major components of Forsythia suspensa showed establishment of Relative correction factor was of good reproducibility by GC and HPLC. No significant differences were observed between the quantitative analysis results of QAMS and ESM in Forsythia suspensa,and the relative error of<5%.Conclusion:The experimental results suggested Shanxi Forsythia suspensa genuineness,which reflected in DNA genetic information,active components and biological effects of antibacterial,antiviral.Due to establishing QAMS method is simple,rapid, sensitive and reproducible characteristics,it can be used for quantitative determination of multi-components in Forsythia suspensa. These study results provide some theory basis for the evaluation of Shanxi genuine and quality of Forsythia suspensa,provide a reference for the pharmacological evaluation and clinical rational drug use of Forsythia suspensa, and provide the scientific basis for Shanxi genuine Forsythia suspensa wildlife tending and standardized planting...