| Objective: Through the identification and research of Euphorbia maculata L.and its related materials Euphorbia thymifolia L.,Euphorbia prostrata Ait.,Euphorbia taihsiensis(Chaw et Koutnik)Oudejians to provide rovide a certain experimental basis for the accuracy of clinical medication.Methods: 1.Using methods such as source identification,character identification,microscopic identification,TLC identification,etc.,the pharmacognosy identification study of Euphorbia maculata L.and its adulterants were carried out.2.A total of 30 samples of Euphorbia maculata L.and its related materials from different places in Guangxi province were collected,and their fingerprints were analyzed and compared by HPLC,and SPSS21.0 software was used to conduct 30 batches of samples.Cluster analysis(HCA),principal component analysis(PCA),establish its cluster analysis dendrogram and principal component characteristic map.3.Collect40 samples of Euphorbia maculata L.and its adulterants from all over Guangxi,extract their genomic DNA,use commonly used DNA barcode sequences rbc L,psb A-trn H for PCR amplification,and use Bio Edit 7.1,MEGA 7.0 for the sequencing results and other software into the analysis.Results: 1.Euphorbia maculata L.and its adulterants are plants of the same family and genus.The original plants have similar morphology.Nonglandular hair can be seen on the surface of leaves and capsules of Euphorbia maculata L.and Euphorbia thymifolia L.Euphorbia prostrata Ait.only has hairs on the edges of the capsules.The whole plant of Euphorbia taihsiensis(Chaw et Koutnik)Oudejians is smooth and hairless;the traits of the four medicinal materials are very similar;the root cross section,stem cross section,leaf vein cross section,leaf upper surface and other characteristics are basically the same,the lower surface characteristics of the leaf: Euphorbia maculata L.and Euphorbia thymifolia L.can be seen nonglandular hair,Euphorbia prostrata Ait.and Euphorbia taihsiensis(Chaw et Koutnik)Oudejians have no nonglandular hairs.Nonglandular hairs can be seen in the powders of Euphorbia maculata L.,Euphorbia thymifolia L.and Euphorbia prostrata Ait.,but not Euphorbia taihsiensis(Chaw et Koutnik)Oudejians.Different medicinal materials are separated by thin layer chromatography different fluorescent spots,and the same fluorescent spots appear on the position corresponding to the control substance.2.Comparing HPLC fingerprints of Euphorbia maculata L.and its adulterants,Samples of the same samples is greater than 0.87.there are obvious differences in the patterns of different samples.The cluster analysis and principal component analysis results are consistent,and the samples are divided.There are four categories,S1～S10 are the first category,which is Euphorbia maculata L.,S11～S18 are the second category,which is Euphorbia prostrata Ait.S19～S24 are the third category,which is Euphorbia thymifolia L.,and S25～S30 are the fourth category.,is Euphorbia taihsiensis(Chaw et Koutnik)Oudejians.3.The sequence length of rbc L and psb A-trn H are 548 bp and 631 bp,GC base content are38.1～52.5%,22.0～23.0%,and the number of conservative sites is 513～516,510～558,respectively.The maximum intraspecific genetic distance of Euphorbia maculata L.was significantly smaller than the minimum interspecific genetic distance between it and its related species.The phylogenetic tree constructed jointly by rbc L and psb A-trn H,the analysis results show that the samples of Euphorbia maculata L.are clustered into a single line(the support rate is 100),and can be clearly distinguished from its related species.Conclusion:It is more difficult to distinguish Euphorbia maculata L.and its related materials by using traits and microscopic methods,while TLC,HPLC,and DNA barcode techniques can effectively distinguish them,but TLC is greatly affected by the factors of the laboratory environment.The results of this experiment can provides rovide a certain experimental basis for the accuracy of clinical medication. |
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