| Objective: In this article,using the HPLC and UV method,berberine hydrochloride and total alkaloids in different parts of cultivated and wild Coptis chinensis were quantitatively determined,the difference of content in the different parts were analyzed.by Coomassie brilliant blue technique and protein SDS-PAGE vertical electrophoresis technology,the protein content of cultivated and wild Coptis in different parts were comparatively analyzed and compared,so as to investigate the biogenetic enzyme and biogenetic pathway in Coptis,and to provide the basis for ensuring the quality and quality control of medicinal plant resources.Methods: After full wavelength scanning of 200-600 nm,the wavelength of 345 nm was measured by ultraviolet spectrophotometry.HPLC was C18(4.6 mm×250 mm,5 μm)chromatographic column,acetonitrile and 0.4% phosphoric acid aqueous solution equal elution,the flow rate was 1.0 mL/min,the injection volume was 10μL,the detection wavelength was 345 nm,and the column temperature was 30 ℃.Through the preparation of berberine hydrochloride affinity medium fishing related metabolic enzymes,using SDS-PAGE gel electrophoresis and Coomassie brilliant blue method,the protein bands of protein abundance,relative molecular weight,A595 and nm per gram of protein content in Radix as the index,the difference of water extraction,Tris-HCl method,ammonium sulfate precipitation method from Coptis chinensis protein and the wild and planting Coptis chinensis,analysis and evaluation of different parts of protein.Results: The amount of total alkaloid in rhizome,stem and root of cultivated Coptis plants are respectively(9.875±0.029)%,(6.406±0.033)% and(1.924±0.037)%,and the amount of berberine hydrochloride in rhizome,stems and roots of cultivated Coptis plant are(0.4505 ±0.0373)%,(0.1673±0.0331)% and(0.0879±0.0293)%.Comparatively,those numbers in wild Coptis chinensis,when it came to the total alkaloid,is(7.953±0.028)% in stems and(0.644±0.031)% in leaves,and when it comes to berberine hydrochloride,the percentages are(0.3802±0.0323)% in stems and(0.1597±0.0315)% in leaves.Preferably,the Tris-HCl method was a better method to extract the optimum Coptis protein as the abundance of the extracted Coptis chinensis protein was the highest,and the protein content per-gram in medicinal materials was also the highest.There were differences of protein discovered in different parts of cultivated and wild Coptis chinensis.Protein abundance and protein content per gram of herbs could be put in order as follows: cultivated Coptis rhizome>wild Coptis rhizome>cultivated Coptis stem> wild Coptis stem and leaf>cultivated Coptis roots.Besides,through the clustering analysis of protein in cultivated and wild Coptis,there was a significant relationship between them in roots while there was no obvious relationship in other parts.Conclusion: There were obvious differences between the alkaloid content in different parts of wild and cultivated plants of Coptis chinensis.The berberine hydrochloride and total alkaloid content in the rhizome of cultivated Coptis were significantly higher than those in the roots and stems.And the content of those in wild Coptis was also higher in the stem than in the leaf.Comparatively,the content in cultivated Coptis was higher than that in wild coptis.As far as the total alkaloid content was concerned,the content of rhizome of Rhizoma Coptidis was the higher and the quality was the better.The optimized Tris-HCl method could be used to extract the protein from Rhizoma Coptidis,and the protein content of rhizome of Rhizoma Coptidis was significantly higher than that of other parts.And the rhizome protein content of Rhizoma Coptidis 6 years old is higher than that of wild rhizome.The difference of protein in different parts of Coptis chinensis laid a foundation for the study of its primary secondary metabolites and metabolic pathways,and provides a basis for the research and quality control of Coptis chinensis germplasm resources. |
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