| This study was carried out systematically to investigate the differences on chemical constituents, inorganic elements, anti-inflammatory efficacy and mechanism of different cultivars of Flos Chrysanthemi, providing some scientific evidences for comprehensively evaluating the qualities of different cultivars of Flos Chrysanthemi and facilitating further refinement and standardization of Flos Chrysanthemi.L9(34) orthogonal design was used to optimize the extraction technology of Flos Chrysanthemi. The best extraction technology was optimized with the the investigate target, peak area of the eight major component of fingerprint in Flos Chrysanthemi and the content of extractum. The extraction process was as follows:ultrasonic extraction method,30-fold ethanol of80%concentration,2times for extraction,40minutes each time. The method is reasonable, feasible and significant to the specification of extraction process and quality control of Flos Chrysanthemi.The results showed that the main chemical compositions had qualitative differences in different cultivars of Flos Chrysanthemi by qualitative analysis of their HPLC fingerprints, UFLC-TOF/MS and GC-MS. Through the establishment of the HPLC fingerprint of different cultivars of Flos Chrysanthemi and the similarity analysis, the clustering analysis and the principal component analysis, the results showed that high similarity between the sample of the same breed, fingerprint common model was stable, reliable and strong specificity. In different cultivars, Boju and Chuju had high similarity, Gongju and Hangju also had high similarity perhaps because they might come from the same species respectively. Through GC-MS qualitative analysis,156kinds of chemical components were identified, including70,58,58,61and76kinds of chemical components were identified in Chuju, Boju, Gongju, Huaiju, Hangju, respectively, all cultivars of the same component for15kinds, and more than3cultivars of the same component for30kinds. The results showed that five different varieties of Flos Chrysanthemi although similar in botany properties, but its small molecule chemical composition and content had significant difference.The results showed that the main chemical compositions had quantitative differences in different cultivars of Flos Chrysanthemi by HPLC quantitative analysis of their main active compounds and ICP-AES analytical method was successfully developed to determine the main trace elements in different cultivars of Flos Chrysanthemi. This validated HPLC method is reliable, reproducible, and valid and could be used to select excellent resources from the cultivars and applied to the intrinsic quality control of cultivars of C. Flos. Besides chlorogenic acid, the content of the other seven active compounds (luteolin-7-O-glucoside,3,5-dicaffeoylquinic acid, apigenin-7-O-glucosid, quercetin, luteolin, apigenin, acacetin) existed significant difference in different cultivars of Flos Chrysanthemi(P<0.05), while there was no significant difference between difference samples for the total content of luteolin-7-O-glucoside and luteolin, apigenin-7-O-glucosid and apigenin. According to the results of quantitative analysis by ICP-AES, the content of the most beneficial elements in Boju and Chuju was higher than other cultivars, but also higher for the potential toxicity elements.The results showed that the anti-inflammatory efficacies of different cultivars of Flos Chrysanthemi groups were different by the ear swelling experiment induced by Xylene and the toes swelling experiment induced by Carrageen of different cultivars of Flos Chrysanthemi. The efficacy of Boju group was best, Chuju group was second. The anti-inflammatory efficacies were of little differences between Gongju, Huaiju and Hangju groups, but poor compared to those of Boju and Chuju ones. In addition, the results indicated that the mechanism of anti-inflammatory was as follows:the extracting solution can inhibit cyclooxygenase pathways, reduce the release of inflammatory mediators such as PGE2and may reduce the production of free radicals during the process of inflammation. |
