Quality Evaluation Study And Variety And QualityRelated Research Of FORSYTHIAE FRUCTUS Based On Chemical And Biological Fingerprint Technology

Objective:To establish the chemical fingerprints and biological fingerprints on FORSYTHIAE FRUCTUS based on the current research method of scientific quality evaluation of Chinese medical materials, evaluate the quality of FORSYTHIAE FRUCTUS from Fructus Forsythiae species identification and chemical qualitative and quantitative identification and other aspects, and establish a new system of identification and quality evaluation in Chinese medicine.Method:Established the biological fingerprints using protein polyacrylamide gel electrophoresis (PAGE), random amplified polymorphic DNA (RAPD) and DNA barcoding technology (to determine the authenticity of species). Established the Chemical constituents fingerprints using ultraviolet spectroscopy (UV), High performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS)(to determine the quality).Appraised the practicability of the chemical and biological fingerprints through comparison of corresponding fingerprints of commercial FORSYTHIAE FRUCTUS and common adulterants. Analyzed the correlation of forsythia biological genetic material and chemical active ingredient, through multivariate statistical analysis method. Analyzed the relationship between the fingerprint data, and established the quality evaluation system through the related research of variety and quality.Results:1. The UV spectra of four kinds of extraction solvent of FORSYTHIAE FRUCTUS were established. The results showed that UV spectra of FORSYTHIAE FRUCTUS from different regions were very close, had the common characteristic peaks, showed that the main component was the same or similar. 2. The HPLC fingerprint with Phillyrin as internal standard reference peak of FORSYTHIAE FRUCTUS was established.20common characteristic peaks were determined, and the common peaks area accounted for more than90%of the total area.3. Chemical compositions of the volatile oil from FORSYTHIAE FRUCTUS were determined by GC-MS analysis.19common peaks were determined, and the common peaks area accounted for more than90%of the total area.4. The FORSYTHIAE FRUCTUS seed PAGE fingerprint was set up.8common bands were determined, and the color depth, width, mobility was similar. The common peaks area accounted for80.9%to96.2%of the total area.5. The results of RAPD analysis showed that polymorphic bands percentage was61.11%, showed genetic diversity.6. A fast identification of official FORSYTHIAE FRUCTUS and its adulterants were achieved, using ITS2sequence, its secondary structure and trnH-psbA sequence.7. Commercial FORSYTHIAE FRUCTUS and fruit of Forsythia viridissima Lindl were analyzed and identified by genuine FORSYTHIAE FRUCTUS chemical and biological fingerprint established in the study. The research method was scientific and practical verified from two aspects of positive and negative.8. Study on relationship between morphological characteristics and the intrinsic quality showed that there had certain relationship between two main effective components and fruit color or the distribution of surface patches. The medicinal material with green and brown and dense patches had better quality.Conclusion:The thesis has preliminary established the fingerprint of multi-dimensional and multi-data on FORSYTHIAE FRUCTUS,created a new system of identification and quality evaluation in Chinese medicine to offer a more accurate and effective way for the identification and quality evaluation on FORSYTHIAE FRUCTUS.