| Yi medicinal materials Boenninghauseniae albiflora Herba and Gaultheriae leucocarpa Herba were the main medicinal materials in the “Yi Medicine cough cranule” and “Yi Medicine Tongfengling”,And Boenninghauseniae sessilicarpa Herba was the same pharmacy method as Boenninghauseniae albiflora Herba Yi Medical clinic in Liangshan area.The Boenninghauseniae sessilicarpa Herba was included in the Yunnan Provincial Standards for Chinese Medicinal Materials(2005 Edition Volume 2,Yi Medicine),Boenninghauseniae albiflora Herba has not been found in other relevant standards.Gaultheriae leucocarpa Herba was one of the main medicinal flavors of Yi medicine preparations such as Green and Kechuan Granule(Z20025849),Yixinkang Capsule(Z20025345),Xiangteng Capsule(Z20025211),Dieda Zhitong Medicine Liquor(Yunnan ZGJF/2005-902),etc.The Gaultheriae leucocarpa Herba was included in the “Yunnan Province Traditional Chinese Medicine Standards”(2005 Edition,Volume 2,Yi Medicine),“Guizhou Province Traditional Chinese Medicinal Materials Quality Standards”(2003 Edition)and “Guangdong Province Chinese Medicinal Materials Standards Third Book”(2019 Edition).There was no recordation about the content determination of Gaultheriae leucocarpa Herba.The quality standard of Gaultheriae leucocarpa Herba was needed to be further improved,which was the main medicinal material in Yi medicine preparation.Based on the above,the following research was proposed.The identification studies of Boenninghauseniae albiflora Herba for instance,identification of original plants,identification of herbs,microscopic identification and TLC etc;extract determination and the routine inspections,which include moisture,total ash,acid-insoluble ash were determined;determination of umbelliferolactone and bergamot etc index components based on HPLC.Formulate the quality standard of Yi medicinal Boenninghauseniae albiflora Herba.Box-Behnken response surface methodology was used to optimize the extraction method of total coumarin from Boenninghauseniae albiflora Herba.The comprehensive quality evaluation of Boenninghauseniae albiflora Herba was studied based on HPLC fingerprint,multi index,one test,multi evaluation and multivariate statistics.Study on the antioxidant activity of the total coumarin extract from Yi medicinal Boenninghauseniae albiflora Herba.Study on the inhibitory effect and mechanism of Boenninghauseniae albiflora Herba and Boenninghauseniae sessilicarpa Herba on the proliferation of human gastric cancer BGC-823 cells in vitro.The identification studies of Yi medicinal material Gaultheriae leucocarpa Herba,were the same as “Boenninghauseniae albiflora Herba”.Study on the content determination of quercitrin based on HPLC.To formulate quality standards for Gaultheriae leucocarpa Herba.Study on the Chemical Constituents and quality evaluation of Gaultheriae leucocarpa Herba.Study on quality of Gaultheriae leucocarpa Herba based on multi-index and multivariate statistical analysis,to provide a research basis for the clinical application and development of Yi medicinal Boenninghauseniae albiflora Herba and Boenninghauseniae sessilicarpa Herba,the results are as follows.1.Study on the quality of Yi medicinal Boenninghauseniae albiflora Herb based on multi-index and multivariate statistical analysis.(1)The cultivars and basic sources of Boenninghauseniae albiflora Herba were determined,and the identification methods of it were established by this study.(2)It was recommended that its moisture content should not exceed 2.57%,total ash content should not exceed 7.65%,acid-insoluble ash content should not exceed1.65%,and alcohol-soluble extract content should not be less than 28.16%.(3)The best extraction method for its total coumarins was 80% ethanol,1:67(g/m L)ratio of material to liquid,ultrasonic-assisted extraction time of 60 min,and ultrasonic assisted extraction temperature 240 W.(4)The UPLC-based fingerprint of Boenninghauseniae albiflora Herba was established.And a multi-index,one-test,multi-evaluation method,which simultaneously use chlorogenic acid,rutin,umbelliferone and isoimperatorin as internal standards to determine eight active ingredients such as neochlorogenic acid,was established in this Study.(5)The content of chlorogenic acid in 11 batches of Boenninghauseniae albiflora Herba samples from different sources is 0.194~3.237mg/g,the content of neochlorogenic acid is 0.273~0.855mg/g,the content of cryptochlorogenic acid is0.359~0.851mg/g,and the content of rutin is 0.880~3.748mg/g,isoquercitrin content is 0.210~2.640mg/g,astragaloside content is 0.145~2.049mg/g,umbelliferone content is 0.372~1.04mg/g,in scopolamine The content of ester is 0.384~1.190mg/g,the content of cervix is 0.151~0.903mg/g,the content of bergamot lactone is0.148~0.274mg/g,and the content of imperatorin is 0.085~0.818mg/g.The content of prehusu is 0.122~0.681mg/g.in content The content of each component in different batches of samples varies greatly.(6)The total content of umbelliferone and bergapten in Boenninghauseniae albiflora Herba was tentatively determined to be no less than 0.072%.(7)According to multi-index and multi-statistical analysis,CHJC-02 is the best sample quality,and the CI indexes of CHJC-08,CHJC-09 and CHJC-11 were very similar.(8)The extracting solution of Boenninghauseniae albiflora Herba showed that they had the significant inhibitory effects on the proliferation of BGC-823 cells(P<0.01),and the relationship between dose-effect and time-effect was obvious.The medium dose of Boenninghauseniae albiflora Herba and high dose of Boenninghauseniae sessilicarpa Herba inhibit the expression of p-AKT and Bcl-2protein more significant than the positive control drug compound Cyclophosphamide.At the same concentration of blood medicine,the inhibition of Boenninghauseniae albiflora Herba was more significant than Boenninghauseniae sessilicarpa Herba.Mechanism of action: Boenninghauseniae albiflora Herba and Boenninghauseniae sessilicarpa Herba inhibit the activation of the Akt signal transduction pathway,thereby down-regulating the expression of p-AKT and Bcl-2 protein to inhibit the proliferation of BGC-823 cells.(9)The quality standards of Yi medicinal,Boenninghauseniae albiflora Herb were formulated,based on the above experimental results.2 Study on the quality of Yi medicinal Gaultheriae leucocarpa Herba based on multi-index and multivariate statistical analysis(1)The origin of Yi medicinal Gaultheriae leucocarpa Herba was determined,and the identification method of Gaultheriae leucocarpa Herba medicinal material was established by this study.(2)In this study,5 compounds were isolated from the 50% ethanol extract of Gaultheriae leucocarpa Herba in the n-butanol extraction site.They are methylsalicylate2-O-β-D-xylosyl(1→6)-β-D-glucopyranoside(Ⅰ),scopoloside(Ⅱ),fraxin(Ⅲ),isoscopoletin(Ⅳ),quercetin(Ⅴ)respectively,among which four compounds,Ⅰ,Ⅲ,Ⅳ,and Ⅴ were isolated from Gaultheriae leucocarpa Herba for the first time.(3)It was recommended that the moisture content of Gaultheriae leucocarpa Herba not exceed 1.71%,the total ash content should not exceed 5.13%,the acid-insoluble ash content should not exceed 0.56%,and the alcohol-soluble extract content should not be less than 11.92%.(4)A method for the determination of 6 active components,such as chlorogenic acid,was established in this study.The HPLC fingerprint of Gaultheriae leucocarpa Herba was established.(5)The content of chlorogenic acid was 0.79~4.15 mg/g,in 10 samples from different sources The content of chlorogenic acid ~0.27 mg/g,1.49 Methyl salicylate2-O-β-D-xylanase(1→6)-β-D-glucoside content 1.65~4.25 mg/g,The content of isoquercetin was 0.11~ mg/g,1.15 The content of amethrin was 0.27~1.16 mg/g,Quercetin was 2.64~11.86 mg/g.in content The content of each component in different batches of samples varies greatly.(6)The results of this study suggest that the content of quercetin in Gaultheriae leucocarpa Herbashould not be less than 0.59%.The results showed that the content of quercetin is not less than 0.59% in Gaultheriae leucocarpa Herba.(7)It showed that the quality of DBZ-02,DBZ-03 and DBZ-01 samples were better than others,There was no significant difference in quality of the three samples,and the quality of DBZ-01 samples was the best,According to the result of multiple indicators and multiple statistical analysis.(8)The quality standard of Gaultheriae leucocarpa Herba was formulated,based on the above test results. |
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