Study On The Establishment Of Quality Standard And Pattern Recognition Of Wannianhao

Objective:Revise the standard of local medicinal materials of Wannianhao in Jilin Province,establish the fingerprint of Wannianhao,and combine the pattern recognition to evaluate the quality of medicinal materials of Wannianhao and the identification of Wannianhao and Aiye and Qinghao and Yinchen.Methods:The DIVA-GIS mapping software was used to the 20 batches of Wannianhao.According to the information of Wannianhao,the geographical distribution map of Wannianhao in Jilin Province was drawn;Qualitative identification of Wannianhao by microscopic method and thin-layer chromatography;According to the"Chinese Pharmacopoeia"2015 edition of the four general rules for the inspection of Wannianhao and the determination of extracts;Using HPLC method,taking the scopoletin as the measurement index,establishing the content determination method and conducting the methodological investigation,and setting the limit;The 20 batches of Wannianhao were tested by HPLC,and the HPLC fingerprint of Wannianhao was established.The chromatographic fingerprint similarity evaluation system of Chinese medicine was used for analysis;Based on the peak area of the common chromatographic peaks of the fingerprints,cluster analysis（HCA）and principal component analysis（PCA）of 20 batches of Wannianhao were carried out using SPSS 22.0 software to establish 20 clusters of Wannianhao cluster tree diagram,and based on the eigenvalues and contribution rate of the principal components,calculate the main component comprehensive score,and comprehensively evaluate the quality of each batch of Wannianhao medicinal materials;Using the chromatographic fingerprint similarity evaluation system of traditional Chinese medicine,the similarity of fingerprints of 18 batches of the Ai Ye,Qinghao,Yinchen with Wannianhao control fingerprint were calculated;Using SPSS 22.0 software,cluster analysis of 20 batches of Wannianhao and 18 batches of Artemisia medicinal materials;Using SIMCA14.0 software,the principal component analysis and orthogonal partial least squares-discriminant analysis of 20 batches of Wannianhao and 18 batches of Artemisia were used to classify and identify Wannianhao and Aiye,Qinghao and Yinchen.Look for the main contribution peaks of the genus Artemisia.Results:Based on the geographical distribution map of Wannianhao,it is confirmed that the geographical distribution of Wannianhao in Jilin Province belongs to a wide span in longitude and latitude,and it belongs to a widely distributed variety in Jilin Province.The plant characteristics and powder microscopic characteristics of Wannianhao were determined.The thin-layer chromatographic method for the identification of coumarins and volatile oils in Wannianhao was established.The method is fast,simple,specific and durable.Comprehensively compare and analyze the test data of each batch of Wannianhao samples,and formulate the detection limit.The moisture should not exceed 13.0%;the total ash should not exceed 8.0%;the acid-insoluble total ash should not exceed 2.0%;the extract should not be less than 20.0%.Taking the scopoletin as an indicator component,there was a large difference in the content of scopoletin in Wannianhao in different producing areas of Jilin Province,and the content ranged from 0.069%to 0.302%.It is tentatively determined that the content of the scopoletin(C₁₀H₈O₄)should not be less than 0.060%.HPLC fingerprints of several batches of Wannianhao from different origins were established,and10 common peaks were identified by similarity software.One of them was identified by the reference of the scopoletin.The similarity of 20 batches of Wannianhao ranged from 0.830to 0.998,and the quality of each batch of medicinal materials varied.The results obtained by cluster analysis are consistent with the similarity results.There is a certain correlation between the cluster analysis results and the sample similarity,and the similarity samples are clustered into one class.Combined with the PCA principal component factor score,the comprehensive evaluation function of Wannianhao was established as follows:F=0.47852F₁+0.19530F₂+0.11841F₃,and the comprehensive scores of 20 batches of Wannianhao were sorted.The S10 had the highest comprehensive score of the main component factor.S19 and S8 have the lowest overall score.The similarity analysis showed that the similarity analysis could distinguish the Qinghao and Wannianhao,while the similarity of the fingerprints of Aiye and Yinchen and Wannianhao existed overlap,and it is difficult to distinguish them from Wannianhao only from the similarity.When cluster analysis was used,four samples of Artemisia were collected into five categories.Qinghao,Yinchen and Aiye are clustered into three group.20 batches of Wannianhao are clustered into two types.Wannianhao with low similarity value is grouped into one category alone.When PCA analysis was performed in two groups,the groups were well separated,indicating that there were differences between groups.The results of OPLS-DA analysis showed that the Wannianhao,Hinghao,Aiye,and Yinchen were all different in the No.7peak,and the peak of No.7 was the main differential marker.Conclusion:Through the systematic research on the quality standard of Wannianhao,the description of the traits,the taste of the medicinal herbs and the indications of the functions of the Wannianhao was improved,and the items of microscopic identification,thin layer chromatography,content determination,moisture,total ash,acid-insoluble ash,and leachate of Wannianhao were added,and the 1977 edition of the quality standard of Wannianhao was revised.Based on fingerprinting technology combined with principal component analysis and cluster analysis,the quality evaluation model of Wannianhao was established.Through the multi-indicator pattern recognition and analysis method of fingerprints,the pattern recognition research of Wannianhao,Qinghao,Aiye and Yinchen was carried out,and their different components were identified.Identification provides a reference basis.