Development Of Fingerprint Of Flos Chrysanthemi And Determination Of Main Flavone Glycosides

Fingerprint comes from forensic sciences, including plant raw materials, drug raw materials and Traditional Chinese Medicine(TCM) , it is used to evaluate the quality of plant raw materials and control quality of TCM and their raw materials, as the diversity of components and their content vary with not only the species but also with the growing conditions, the season when plants are harvested, the process methods and storage duration. Moreover, in the medicine plant production, it is not always necessary to produce pure preparations. Herbal drugs are then standardized to selected component to assure comparability of different preparations of plant origin (herbal teas, extracts, extract evaporates, preparations, formulations, decoctions). So Fingerprint is also used to control quality of TCM and their raw materials as stated by the Chinese Pharmacopoeia, and it has been accepted by WHO as a methodology for the assessment of herbal medicines. Conventional research focuses mainly on the determination of the most active or hazardous components, while fingerprint can offer integral characterization of a complex system with a quantitative degree of reliability and gained more and more attention.Flos Chrysanthemi, anthotaxy of Chrysanthemum morifolium Ramat. is used as a heat-clearing and detoxication herb. And it is a a rich source of secondary compounds (including luteolin -7-O-β-D-glucosid;apigenin -7-O-β-D-glucoside and acacetin -7-O-b-D-galactopyranoside )with a variety of biological activities. For example. Itcan also inhibit the agglutination of blood platelet and promote the myocardial blood circulation and white cell phagocytosis;thus it has been used to treat many diseases such as furuncle and skin nodules. Recently, it has been found to show inhibitory activity against nitric oxide (NO) production in lipopolysaccharide-activated macrophages, and found that acacetin- 7-O-b-D-galactopyranoside which is a compound in it was active in an anti-HIV assay. In order to ensure authenticity, quality, safety and efficacy of the Flos Chrysanthemi, CME, tablet of CME and Flos Chrysanthemi of different sources, grades, processes and storage time.Fingerprint were analyzed by using HPLC and IR which can be used identifying methods for Flos Chrysanthemi, and also be used to control quality of CME and tablet of CME1. Development of HPLC-fingerprint for the quality assessment of Flos ChrysanthemiObjective: To develop the HPLC-fingerprint of Flos Chrysanthemi,CME and tablets of CME.Methods: HPLC-DAD was used to analyze the flavonoids, the main composition of Flos Chrysanthemi,CME and tablets of CME. Agilent Zorbax SB Cih column (250mm x4.6mm, 5um) was used as analysis column, the mobile phase was acetonitrile / pH2.0 phosphate buffer solution with gradient elution. CME,tablets of CME and Flos Chrysanthemi of different sources, grades, processes and store time were analyzed by the described method.Results: By cluster analysis and similarity calculation, twenty-three batches of Fols Chrysanthemi were differentiate from others, and the similarity of them were 0.95-1.00, but the similarity of the other thirteen species was less than 0.90. Among the ten batches of CME, the HPLC-fingerprint and their chemical components were similar to each other;and the similarity of four batches were not obviously different (>0. 96);moreover, the similarity of Flos Chrysanthemi, CME and tablets of CME were more than 0.95, so their chemical components were similar to each other. Conclusion: HPLC fingerprint can be an identifying method for Flos Chrysanthemi and can be used to control quality of CME and tablet of CME .2. The study of fingerprint infrared spectra and clustering analysis on FlosChrysanthemi samplesObjective: To develop IR fingerprint of Flos Chrysanthemi.Methods: On the basis of infrared spectrum fingerprint, principal component analysis and clustering analysis were used to classify Flos Chrysanthemi.Results: ecause of the greater difference between the spectra of Fols Chrysanthemi and the others, the outcome of the clustering was ideal. Among the thirty-three batches of Fols Chrysanthemi, the spectra and chemical components of them were similar to each other, which could be considered as the criterion of evaluating the quality of Fols Chrysanthemi. In order to increase the clustering accuracy, the number of the samples should be increased and also these samples should be more characteristic.Conclusion: On the whole, with the combination of the fingerprine infrared spectrum and the clustering analysis, the genuineness of herbal medicines can be discrinated rapidly.3. Determination of flavonoids and main flavone glycosides in Flos Chrysanthemi and observation of factors influenced the contentsObjective: To compare the content of flavonoids, luteolin-7-O-P-D-glucoside and apigenin -7-O-J3-D-glucoside in Flos Chrysanthemi from different collection time, sources, grades and processes.Methods: The contents of flavonoids was determined by colorimetry, while the contents of luteolin -7-O-P-D-glucosid and apigenin -7-O-p-D-glucoside were determined by RP-HPLC. Agilent Zorbax SB C18 column (250mm x4.6mm, 5pm) was used as analysis column, the mobile phase was acetonitrile / pH2.0 phosphate buffer solution with gradient elution. The flow rate was l.OmLmin-1, the detector was set at 338nm.Results: The contents of flavonoids, luteolin-7-O-p-D-glucoside and apigenin -7-O-P-D-glucoside changed at some degree in Flos Chrysanthemi from different collection time, different plant sites or with different grades, while the contents varied obviously among Flos Chrysanthemi from different source and different sorts. No obvious difference was found in Flos Chrysanthemi from different year.Conclusion: The contents of flavonoids, luteolin-7-O-P-D-glucoside and apigenin -7-O-p-D-glucoside were influenced by process, plane site, source and sorts, especially by source and sorts.