| Objective With the deepening modernization of traditional Chinese medicine(TCM)and the continuous improvement of the quality testing methods of TCM,the cultivation of Bletillae rhizoma across our country is showing a trend of scale and expansion.There is a significant difference in the quality of B.rhizoma from different regions,and there is no comprehensive evaluation standard for B.rhizoma currently.Therefore,this study aims to establish a rapid and accurate UPLC detection method for B.rhizoma from Zhejiang province,and evaluate and analyze the quality of B.rhizoma from different regions.At the same time,an efficient separation and purification process of militarine will be optimized.Finally,a preliminary exploration will be conducted on the anti-photoaging function of B.rhizoma,laying a foundation for the subsequent quality evaluation of B.rhizoma and the application of key natural active ingredients.Methods 1.A rapid,efficient,and accurate UPLC detection method for the content determination of natural active ingredients of B.rhizome were established,and methodological studies were conducted.2.UPLC fingerprints of B.rhizoma from five places of origin including Zhejiang,Guizhou,Gansu,Yunnan and Anhui were constructed,and similarity evaluation and chemometric analysis were carried out.UPLC-Q-TOF/MS was used to detect the metabolic components of B.rhizoma and explore the differences in the composition of B.rhizoma from different origins.3.By using macroporous adsorption resin column chromatography and normal silica gel column chromatography,the solvent and separation gradient were optimized to establish a set of rapid separation and purification technology of militarine.4.Based on network pharmacology,the target network and PPI network for the components of B.rhizoma and skin photoaging was explored,and GO and KEGG pathway analysis and molecular docking were conducted,to explore the action mechanism of B.rhizoma against skin photoaging.Results 1.A UPLC method for the determination of the components of B.rhizoma was established,achieving better detection results for the complex component system of B.rhizoma,with faster analysis speed,higher sensitivity,and better separation,and the methodological validation shows good results..2.The UPLC fingerprints of B.rhizoma from Anhui,Gansu,Guizhou,Yunnan,and Zhejiang was established,33 common peaks were found,and a total of 66 components were identified through UPLC-Q-TOF/MS.The content of components in B.rhizoma from different origins varies greatly,among which the contents of phenanthrene,bibenzyl and ester were higher.The high content of militarine in B.rhizoma from Zhejiang proves that it can be used as a high-quality raw material in the production and application of militarine.3.Extraction segmentation of B.rhizoma with n-butanol were separated,from which militarine were easily and quickly separated,resulting in a final yield of 0.067%.4.Network pharmacology predicted that the militarine in B.rhizoma form 13 stable hydrogen bonds with ESR1,forming a hairpin tight structure that inhibits ESR1.Through the estrogen signaling pathway,it inhibits melanin synthesis and tissue fibrosis,exerting an anti-skin photoaging effect.Conclusion The contents of B.rhizoma components from different producing origins were significantly different,among which phenanthrene,bibenzyl and ester components were the main different secondary metabolites.The content of militarine is relatively high in B.rhizoma from Zhejiang.A rapid and efficient separation and purification of militarine was achieved using macroporous adsorption resin and normal phase silica gel column chromatography.A purity of 96.48%militarine was obtained from the 30%ethanol elution fraction of the n-butanol layer of B.rhizoma from Zhejiang,with a final yield of 0.067%.Network pharmacology analysis predicts that the anti-skin photoaging effect of B.rhizoma may be mainly due to its inhibitory effect of militarine on ESR1,which inhibits melanin synthesis and tissue fibrosis through the estrogen signaling pathway. |
PDF Full Text Request