Study On Identification And Growth Year Of Mountain-cultivated Ginseng Based On LC-MS Technology

Objective:Mountain-cultivated ginseng is famous as a precious traditional Chinese medicine.In recent years,with its common application and development in food and medical areas,its quality control has become the focused issue.Because its scarce resource,high price and large demand,fake products flood in the market.It is particularly important to formulate scientific and accurate quality control methods for mountain-cultivated ginseng and to improve the quality of ginseng.As a perennial plant,harvest time is an important index for the quality of the ginseng.Thus correct judgment of ginseng'sage is of great significance toits quality control.In this paper,we focus on investigating mountain-cultivated ginseng grew in???????.The HPLC fingerprint of ginseng was combined with multi-component content determination method.LC-MS was used to analyze the chemical constituents of mountain-cultivated ginseng.Based on metabonomics,the metabolites of mountain-cultivated ginseng were studied in depth,combined with multivariate statistical methods.The results were analyzed,which has certain significance for the quality evaluation,the age judgement and strain identification of mountain-cultivated ginseng.Method:?1?The optimal extraction conditions of mountain-cultivated ginseng were determined by single factor investigation on the extraction method of mountain-cultivated ginseng and the final extraction conditions were determined by uniform design optimization.The chromatographic conditions of the samples were investigated by HPLC-PDA,and the HPLC fingerprint of mountain-cultivated ginseng was established.?2?HPLC-PAD was used to analyze the fingerprint of mountain-cultivated ginseng,and the cdf file was imported into the Chinese Pharmacopoeia Committee published the"Chinese medicine chromatographic fingerprint similarity evaluation system A version"?2012 version?software for analysis,generating three Fingerprints of mountain-cultivated ginseng under different ages and the standard map of undergrowth ginseng produced by all ginseng.UPLC-Q-TOF/MS was used to identify the common peaks of fingerprints.Cluster analysis and orthogonal partial least squares analysis were performed using SIMCA-P software with peak area as the variable.?3?UPLC-Q-TOF/MS-based plant metabolomics analysis of the forest ginseng and garden ginseng,research on 30 batches of mountain-cultivated ginseng and 10batches of cultivated ginseng differential metabolites.Result:?1?The optimal extraction process of mountain-cultivated ginseng was determined:the extraction method was ultrasonic extraction,and then re-dissolved by rotary evaporation to dryness;the extraction solvent was 70%methanol;the extraction time was 100 min;and the extraction temperature was 50°C.The fingerprint method of mountain-cultivated ginseng combined with multi-component content determination method was established:the column was Waters CORTECS C₁₈?150 mm×4.6 mm,2.7?m?,the detection wavelength was 203 nm,the column temperature was 30°C,the injection volume was 5?L,and the flow rate was 1.0m L/min.The mobile phase was eluted with 0.1%formic acid-acetonitrile as phase A and 0.1%formic acid-water solution as phase B.The contents of ginsenosides Rg₁,Re,Rb₁,Rc and Rd were determined.?2?The standard fingerprint of mountain-cultivated ginseng was generated.The ginsenoside Re was used as the reference peak,and 33 common peaks were calibrated.Sixteen peaks were analyzed by UPLC-MS.The OPLS-DA discriminant model established by the common peak area of the fingerprint map can distinguish mountain-cultivated ginseng from participating in the different ages?10,15,and 20years?.?3?UPLC-Q-TOF/MS-based plant metabolomics analysis mountain-cultivated ginseng participated in cultivated ginseng,using PCA,PLS-DA,OPLS-DA and other analytical methods to accurately distinguish mountain-cultivated ginseng from different ages,and to identify mountain-cultivated ginseng participated in cultivated ginseng,and analyze the abundance of different metabolites with different contribution rates.Conclusion:In this paper,the fingerprint method of mountain-cultivated ginseng combined with multi-component content determination method was established,and the quality of mountain-cultivated ginseng was evaluated.Using the fingerprint map combined with the mathematical statistics method,the OPLS-DA discriminant model was established,which can be used to distinguish the ginseng under different ages,mountain-cultivated ginseng and cultivated ginseng.Plant metabolomics based on UPLC-Q-TOF/MS distinguishes mountain-cultivated ginseng under different ages,mountain-cultivated ginseng and cultivated ginseng from the metabolite level technology.The detection method established in this experiment is reasonable,stable,reliable and reproducible.It can be used for the quality evaluation analysis of ginseng under different ages and the identification of mountain-cultivated ginseng.