| Purpose1 The determination of amino acids in different kinds of animal medicinal materials was carried out in order to provide the experimental basis for qualitative analysis of nine kinds of medicinal materials such as leech,Bombyx bassiana,hillbilly worm,centipede,scorpion,gecko,Agkistrodon acutus and snake from the point of view of amino acid species.2 The contents of 18 kinds of amino acids in nine kinds of medicinal materials of leech,Bombyx bassiana,hillbilly worm,centipede,scorpion,gecko,Agkistrodon acutus and snake were determined,and the similarities and differences in kinds and contents of amino acids were compared and analyzed.3 The quality of leeches and leeches were evaluated by grey correlation quality evaluation model.4 By clustering analysis,9 kinds of medicinal materials,such as leech and leech,were classified into several groups,and the quality differences among them were analyzed.5 Stepwise regression analysis was used to select the most significant factors affecting the total amino acids in leeches,leeches,etc.in order to optimize the detection index of amino acids in quality control.6 According to the content of amino acids,the species of leeches and leeches in annellids,silkworm,hillbilly,centipede,scorpion,gecko,Agkistrodon acutus and serpent were analyzed.Materials and methods1 Materials: 9 batch of leech medicinal herbs,13 batches of commercial medicinal materials of leech,9 batches of commercial medicinal materials of silkworm,10 batches of medicinal herbs of woodlouse insect,8 batches of medicinal herbs of centipede,10 batches of commercial medicinal materials,8 batches of commercial medicinal materials of gecko,10 of medicinal products of Agkistrodon acutus,10 of medicinal products of snake.2 Method2.1 Qualitative Analysis and content determination of Amino acids in Hirudo,Leech and other Commercial Medicinal Materials HPLC method was used to determine the amino acids in leeches and leeches by pre-column derivatization and gradient elution.2.2 Establishment of grey relational degree quality evaluation model for commercial medicinal materials such as leech,etc.The data set of grey relational degree model is composed of the total amino acid content data of leech and leech.According to the calculation requirement and formula of grey correlation degree,the correlation number,correlation degree and relative correlation degree RI of the relative optimal and the worst reference sequence of each sample were calculated respectively,and the grey correlation degree quality evaluation model of commercial medicinal materials of antelope antler and deer antler was established.(1)Raw data normalized:Yik=Xik(?)k(2)The best coefficient of incidence:(?)(3)The worst coefficient of incidence:(?)(4)The best incidence degree:(?)(5)The worst incidence degree:(?)(6)Relative incidence degree ri:(?)2.3 Systematic cluster analysis of commercial medicinal materials such as leech and leech SPSS19.0 statistical software was used to analyze the data systematically.2.4 Multivariate correlation analysis of commercial medicinal materials such as leech Leech was carried out by using SAS or SPSS statistical analysis system and statistical analysis software.The data collected from the sample data sets of various animal medicinal materials were analyzed by multivariate correlation analysis.Secondly,multiple regression analysis was used to establish the multivariate relationship equation among principal components.Y=x1+B+X++x3+Dx4+...N+k The coefficient X,A,B,C and D of each independent variable in the equation.Positive and negative..N showed positive and negative correlation with dependent variable Y respectively.The positive correlation between the dependent variable and the independent variable indicates that the dependent variable can indicate the change of the independent variable,that is,it can simplify the detection of the independent variable in theory and replace it with the dependent variable;on the contrary,the negative correlation between the dependent variable and the independent variable indicates that the dependent variable can not indicate the change of the independent variable,that is to say,it can not be replaced by the dependent variable.Detection of the independent variable,that is,the independent variable is the index that must be detected.2.5 Stepwise discriminant analysis of commercial medicinal materials,such as leech,etc.The original variables were selected for stepwise discriminant analysis.Wilks and Lambda data were used to test the statistical significance of the discriminant function.According to the results,defective amino acid variables were eliminated.Result1 Except for scorpion and snakes containing 17 common pattern peaks,leeches,leeches,Bombyx silkworms,woodlouse worms,centipede,Gekko and Agkistrodon have 18 common pattern peaks.2 The amino acid content of more than ten kinds of animal medicinal materials,such as leech,leech and silkworm,showed that amino acid is an essential component in animal body.Among the varieties and specifications,17 kinds of amino acids and 8 kinds of essential amino acids are the lowest.The contents of all kinds of amino acids are different,but the total amount of amino acids accounts for the body weight.About 50%,the essential amino acid content is about 20%.3 The grey correlation quality model was established by leeches,Bombyx bassiana and so on,but all the varieties were close to the ri value of 0.5,but the range of ri change was not significant.4 The cluster analysis of different species of animal medicine was carried out by spss19.0software.Using Hierarchical Cluster Analysis,Euclidean distance as similarity measure between samples and 18 kinds of amino acids as variables,the similarity degree of animal medicinal materials from the same species and different commercial places is different,which can be clustered into multiple groups.5 The relationship between 18 amino acids in different animal medicines revealed by multiple regression equation shows that the content of one or more of them can be used to indicate the amount of other components,and the content of several components can be calculated simultaneously by detecting one index,such as the total amino acid content of leech medicines by determining methionine.The content of total amino acids can be obtained by measuring aspartic acid and methionine.6 According to the origin of medicinal materials,nine kinds of medicinal materials are divided into three categories: annelids,arthropods and reptiles.The accuracy of discriminant analysis was 100%.Conclusion1 The amino acids in leeches,leeches and Bombyx bassiana were compared and analyzed by HPLC,and it was found that the species of amino acids could be used to qualitatively analyze animal medicinal materials.2 In this experiment,the content of 18 amino acids in leech,leech,horse and silkworm was determined by high performance liquid chromatography with gradient elution and pre-column derivatization.The method is simple,rapid and reproducible.The contents of the 18 amino acids can be used as indicators for the quality evaluation and control of different species of animal medicines.3 The results of grey correlation quality evaluation showed that there was no significant difference in the quality of animal medicinal materials between the same species and different commercial areas.4 Using amino acid as index,cluster analysis can accurately cluster animal medicinal materials from different producing areas.5 By using stepwise regression method and amino acid as index,18 kinds of amino acids in different species of animal medicinal materials can be simplified and tested,and the simplified indexes of amino acids of different varieties are also different.6 By discriminant analysis,the contents of amino acids can be accurately distinguished from leeches and leeches of annelids,Bombyx hillbilly,centipede and scorpion from arthropods,gecko,Agkistrodon acutus and snake from black shoot. |
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