| Objective: In this paper,by constructing a Liquid Chromatography fingerprint of Stephania tetrandra and analyzing the differences in the content of fangchinoline and tetrandrine in the main effective ingredients of Stephania tetrandra from different places,establish a Stephania tetrandra identification and quality evaluation method to evaluate the quality of Stephania tetrandra from different places;By exploring the correlation between environmental factors such as geography,climate,soil,phenotype,traits of medicinal materials and the content of active ingredients,and reveal the relationship between external factors and the quality of medicinal materials,and provide a basis for identification of Stephania tetrandra varieties,germplasm classification,selection of excellent germplasm,and selection of the best place of origin.Methods:(1)HPLC method was used to determine the contents of main effective components of fangchinoline and tetrandrine and to construct UPLC fingerprint;(2)The use of the network search information to obtained the climatic factors of various areas,intelligent handheld GPS to obtain altitude and longitude and latitude;(3)Determine the different nutrient elements of soil with reference to the "Forestry Industry Standard of the People’s Republic of China";(4)Phenotypic traits and medicinal material traits were observed by visual inspection and tape measure.SPSS was used to analyze the variance of the contents of Stephania tetrandra active components from different sources to evaluate the quality difference;The R language software was used for cluster analysis of medicinal materials from different areas to obtain dominant population and select excellent strains.SPSS was used to analyze the correlation between external factors such as environment,soil,phenotype,medicinal material traits and the content of active ingredients,and establish multiple linear regression equations to obtain the factors with significant correlation with the content.Results:(1)Established a common pattern of UPLC fingerprint of Stephania tetrandra,and 8 common fingerprint peaks were demarcated,and The similarity of UPLC fingerprints from different producing areas was compared,the results showed that the similarity was greater than or equal to 0.90,the similarity was good,and the quality of the tested medicinal materials was stable.(2)The sum of the contents of fangchinoline and tetrandrine in the chemical constituents of Stephania tetrandra medicinal materials from different origins(hereinafter referred to as the total content)is higher than the pharmacopoeia standard(not less than 1.6%),and the medicinal materials of all origins are qualified.According to the results of variance analysis among difference regions,the content of Guangdong is the highest among the six provinces,followed by Fujian,Jiangxi,Anhui and Hunan,and Zhejiang is the lowest.According to the results of cluster analysis,38 counties can be grouped into 4 groups.The first group contains 17 counties,with a total content averaged of 2.20%,which is the lowest among the four groups.The second group is Shixing County,Guangdong Province,with only one county,with a total content averaged of 4.66% and the highest content;The third group contained 8 counties with a total content averaged of 2.81%,ranking the third;The fourth group contained 12 counties with a total content averaged of 3.35%,ranking the second.According to the comprehensive comparison,Shixing county of Guangdong and Shitai county of Anhui,Guixi city of jiangxi,Poyang county of Jiangxi,Dongzhi county of Anhui,Duchang county of Jiangxi,Shunchang county of Fujian,Xinjian district of Jiangxi,Xiangxiang city of Hunan,Quannan county of Jiangxi,Shexian county of Anhui,Songhou county of Fujian and Nanxiong city of Guangdong are the excellent populations in the second group of Stephania tetrandra.(3)The clustering analysis results of 417 strains of Stephania tetrandra showed that 417 strains of Stephania tetrandra could cluster into 4 groups,and the first group contained 173 strains,with a total content averaged of 1.84%,which was the lowest among the four groups.The second group contained 71 strains,with a total content averaged of 3.78%,ranking the second;The third group contained 34 strains,with a total content averaged of 5.40%,ranking the first;There are 139 strains in the fourth group,with a total content averaged of 2.69%,ranking third.Based on the comprehensive cluster analysis results,it can be seen that excellent strains are mainly clustered in the third group.(4)The results of the correlation analysis between the content of main effective components and the geographical and climatic factors showed that the latitude,annual average temperature and annual rainfall were significantly correlated with the content of main effective components.Among them,there was a significant negative correlation between fangchinoline and latitude,and the content of fangchinoline was higher at low latitude(about 24 degrees north latitude);There was a significant positive correlation with the average annual temperature,and the content of the high average annual temperature(about 18℃)was higher.Tetrandrine was negatively correlated with the annual rainfall,and the annual rainfall was about 1400 mm.(5)The fangchinoline was positively correlated with exchangeable calcium and available zinc,and positively correlated with available boron;Tetrandrine was positively correlated with exchangeable magnesium;Total content was positively correlated with exchangeable calcium,exchangeable magnesium,available zinc and available boron.The results of regression analysis showed that exchangeable calcium and available boron were the main soil factors affecting the content of Stephania tetrandra.(6)Through the study on the correlation between the main effective components of Stephania tetrandra and phenotypic traits,it was found that the main effective components of fangchinoline were significantly correlated with leaf shape,leaf veins,stem color and stem thickness,but there was no significant correlation between the content of tetrandrine and phenotypic traits.(7)Through the research on the correlation between the content of the main active ingredient of Stephania tetrandra and the properties of the medicinal materials,it was found that the content of the main effective components was significantly correlated with the texture of the medicinal materials,and the content of those with a solid texture was lower.It was significantly correlated with the surface color,and the content of the surface color was higher.It was significantly correlated with the color of the section,and the content of light white was lower.The higher the powdery,the lower the content.The results are inconsistent with the results of the traditional quality evaluation standards of Stephania tetrandra medicinal materials,and the reasons need to be further studied.Conclusion: The established UPLC fingerprint of Stephania tetrandra identified eight common peaks,and the similarity evaluation results were consistent with the requirements of the establishment of liquid phase fingerprint.The results of the evaluation on the quality of Fangfang from different origins effectively screened out the dominant population and excellent strains of Stephania tetrandra;Based on the correlation analysis of the content of active ingredients and the environmental factors of the origin producing area of Stephania tetrandra,the geographical and environmental factors affecting the content of active ingredients were obtained.Based on the correlation analysis of the content of the active ingredient and the Stephania tetrandra phenotype and the properties of the medicinal materials,the phenotypic and medicinal properties related to the content of the active ingredients were clarified.The results of this paper can provide experimental basis for the selection of superior Stephania tetrandra population and superior germplasm,the selection of suitable production environment,and the prediction of high content germplasm through phenotype and medicinal material characteristics. |
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