| Rationales:Hippocampus is an precious marine animal medicinal material,it has a wide range of remarkable curative effects,like invigorating kidney yang warmly,dispersing swelling,anti-oxidation,anti-aging and so on,because of this,it also has the reputation of"South ginseng".In recent years,with the recognition of the pharmacodynamics about hippocampus,the catch of it has increased clearly,together with the change in marine ecological environment have caused the resource shortage of hippocampus in the sea area of our country,and the medicinal materials in the market are mostly imported from foreign country now.As a result,the market resources of hippocampus have become chaotic.The variety of hippocampus sold in market is not only the five breeds in the Pharmacopoeia of the people's Republic of China,but also some adulteration of others,so is there a quality difference between different breeds of hippocampus?The price in two weight grades has huge difference,the price of hippocampus in light weight about3.5 g is 5,500 yuan/kg,the price in heavy weight about 10.8 g is about 10,000 to 12,000 yuan/kg,so what are the differences in the physical basis of the hippocampus at different grades,and are the market price differences reasonable?And does the processing technology affect the chemical composition of hippocampus?These are all the quality control problems that hippocampus are facing.Metabolomics techniques collect big data through the detection of sample composition,and use the data to compare the differences and evaluate the quality among different specifications of medicinal materials after multivariate statistical analysis has its unique technical advantages.Therefore,the methods of metabolomics were used to compare and analyze the ingredients of different commercial medicinal materials in hippocampus has its great significance in evaluating the quality.Objective:Compare the chemical composition of hippocampus in different breeds,grades and processing technology to screen the different substances in different groups and evaluate the quality of different commercial medicinal materials in hippocampus,then take suggestion for the selection of hippocampus varieties to be used as medicine,investigate the rationality of the price variance of different grades hippocampus,and explore the scientific basis for the processing of medicinal materials.Methods:In this study,a suitable method for the detection of ~1H-NMR and GC-MS in hippocampus was obtained by the exploration in the early stage,and then determine the components of different commercial medicinal materials in hippocampus.By comparing the spectrum,we observed the visual difference of hippocampus in different groups.After processing the spectrum synchronously,the content of components was compared by using the technique of metabolomics analysis and SIMCA-P software,etc.In combination with the results of single factor variance analysis of relative peak area among groups,the compounds with large contribution rate to the difference between groups were selected as representative substances to cause separation between groups,and the functional effects of different substances were analyzed to evaluate the quality difference of hippocampus with different commodity specifications.Results:1.Through the exploration and improvement of the method of sample preparation and the condition of instrument detection,the ~1H-NMR fingerprinting and GC-MS spectrum of hippocampus were established,and33 and 42 kinds of chemical constituents were identified in spectra,respectively,both of them were used to compare the compounds of different commercial medicinal materials in hippocampus in subsequent experiments.2.The difference of chemical composition in hippocampus of different breeds was compared by using NMR metabolomics approach.It was found that there were great differences between individuals in Hippocampus histrix Kaup and Hippocampus japonicus Kaup,in which the content of isoleucine,leucine,valine,alanine,glycine,tyrosine,phenylalanine,lactic acid,acetic acid,choline,glycerol and fumaric acid were different.Combined with multivariate statistical analysis,it showed that the content of glutamic acid,aspartic acid and glycerol in Hippocampus kelloggi Jordan et Snyder were significantly higher than those in Hippocampus kuda Bleeker,while the other chemical components were similar;and that Hippocampus trimaculatus Leach compared to Hippocampus kuda Bleeker,the content of isoleucine,leucine,arginine,methionine,glutamate,hydroxyproline,proline and cysteine decreased markedly,while acetic acid,taurine,glycine and glycerol increased.Using GC-MS metabolomics approach to compare the chemical composition,there was no significant differences among Hippocampus kelloggi Jordan et Snyder,Hippocampus trimaculatus Leach,Hippocampus japonicus Kaup and Hippocampus kuda Bleeker;compared with Hippocampus histrix Kaup and Hippocampus kuda Bleeker,many fatty acids,like:tetradecanoic acid,hexadecanoic acid and the steroid composition of androst-4-ene-3,17-dione had decreased in Hippocampus histrix Kaup;while the content of xylitol,octadecanoic acid and arachidonic acid on a high level.3.Comparison of chemical constituents in different grades of Hippocampus kuda Bleeker by ~1H-NMR technique,the heavy hippocampus compared with the light one has increased in glycerol and acetic acid,while lactate,creatinine,tyrosine decreased significantly;by GC-MS technology,the result showed that the difference between the two grades is weak,but the cholesterol content of cholesterols is stable,up to 90%.4.After processing,the texture of Hippocampus kuda Bleeker become crispy with a unique flavor.Comparison of chemical constituents on Hippocampus kuda Bleeker before and after processing by ~1H-NMR technique,showed that it had significantly increased in the content of some chemical components after processing,including arginine,isoleucine,valine,aspartic acid and taurine,and the content of glutamic acid,ornithine,proline and some other components had decreased;by GC-MS technology,the content of some fatty acids and liposoluble components decreased after processing,suggesting that chemical transformation between fatty acids and amino acids may exist in the process.The whole elements analysis in Hippocampus kuda Bleeker before and after processing were detected by ICP-MS technique showed that the relative content of elements fluctuated slightly,but had no statistical significance.Conclusion:1.Compared the chemical constituents of hippocampus in different breeds with different detection techniques,it was found that there were great differences between individuals in Hippocampus histrix Kaup and Hippocampus japonicus Kaup,Hippocampus kelloggi Jordan et Snyder and Hippocampus kuda Bleeker had high level of various functional amino acids and steroids,and the content of individual components is stable,so they are more suitable to be choosed as Chinese medicine.And it is scientific and reasonable to use Hippocampus kuda Bleeker in Guilingji.If Hippocampus kuda Bleeker face to a shortage of resources,the Hippocampus kelloggi Jordan et Snyder can be used as an alternative.2.Compared the chemical constituents of hippocampus in different grades with different detection techniques,it was found that there were few different components between different grades hippocampus,it is necessary to carry on the quantitative analysis about the composition content to perfect the quality evaluation of the two grades,and draw a feasible conclusion.3.Compared the chemical constituents of hippocampus before and after procssing with different detection techniques,after processing,the texture of Hippocampus kuda Bleeker became fragile,and the difference of individual components had decreased,the content reached a more uniform and stable state,which showed that the processing technology of hippocampus had reasonable scientific basis.In addition,there may be the decomposition of components or chemical transformation between components had occured in the process,but the specific mechanism of change needs to be further explored.However,the processing technology of hippocampus had little effect on the content of inorganic elements. |
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