Systematic Characterization And Comparison Of Garden-cultivated Ginseng And Mountain-cultivated Ginseng Based On LC/MS Technique

Objective: Ginseng can be divided into three categories according to its growing environment and cultivation methods,namely garden-cultivated ginseng,mountain-cultivated ginseng and wild ginseng.In 2008,the mountain-cultivated ginseng was included in the category of wild ginseng in“Wild Ginseng Identification and Grading Quality”(GB/T18765-2008)promulgated by the state.Due to the different growth environments and years,the quality and efficacy of ginseng are different,and the degree of market recognition also varies.Mountain-cultivated ginseng is much more expensive and also in great demand,while the current research on the chemical composition and quality control of the mountain-cultivated ginseng is still scarce.The Chinese Pharmacopoeia divides ginseng into garden-cultivated ginseng and mountain-cultivated ginseng.The lack of specific quality standards for different sources of ginseng makes it difficult to identify and distinguish these ginseng samples.Therefore,it is of great significance to explore the chemical composition of mountain ginseng(including both wild and cultivated ones)and effectively distinguish garden-cultivated ginseng and mountain-cultivated ginseng to provide scientific evidence for elaborating the scientific quality standard of mountain-cultivated ginseng.Methods: 1.An off-line two-dimensional liquid phase separation system based on HILIC × RP was constructed,and mass spectrometry data were acquired in both positive and negative ion modes to targeted characterize saponins in mountain-cultivated ginseng.2.A quantitative/semi-quantitative method based on the UHPLC-QtrapMS dynamic multi-reaction monitoring method was established,which realized the quantitative analysis of 14 saponins and the semi-quantitative analysis of 199 components.Multivariate statistical analysis was used to analyze the differential metabolites of garden-cultivated ginseng and mountain-cultivated ginseng.3.Using UPLC-Q-TOF/MS technology,positive and negative ion modes were used for mass spectrometry scanning.The chemical constituents in the standard decoction of ginseng dispensing granules were analyzed by database searching,reference substance comparison and MS DIAL software-assisted analysis.The fingerprint of ginseng dispensing granules was established by ultra performance liquid chromatography,and the methodology was verified.Results: 1.According to mass spectrometry data of mountain-cultivated ginseng in both positive and negative modes,559 ginsenosides were finally tentatively identified from mountain-cultivated ginseng,including 148 PPT type,191 PPD type,40 OA type,17 OT type,124 C17-side chain variants and39 others.2.A quantitative / semi-quantitative method based on the UHPLC-QtrapMS dynamic multi-reaction monitoring method was established.The results were acceptable and the sensitivity of the method was good.This method was applied to the analysis of 15-year-old,20-year-old and 25-year-old mountaincultivated ginseng and garden-cultivated ginseng.PCA,OPLS-DA and other methods were used to distinguish mountain-cultivated ginseng and gardencultivated ginseng and mountain-cultivated ginseng of different years.3.A total of 57 compounds,including 25 saponins,12 amino acids and their derivatives,5 bases and nucleosides,together with 15 other compounds,were tentatively identified in the standard decoction of ginseng dispensing granules.The HPLC fingerprint of the ginseng dispensing granules was established with 8 common peaks.The precision,stability and reproducibility of the HPLC fingerprint were acceptable.The components in different batches of ginseng dispensing granules from different manufacturers were similar,and only slight differences existed in contents.Conclusion: an offline two-dimensional method was established to systematically analyze the chemical components in mountain-cultivated ginseng.A multi-component quantitative analysis method was developed and applied to garden ginseng and mountain-cultivated ginseng of different ages.The comparison and analysis of mountain-cultivated ginseng of different ages were carried out to lay the foundation for the quality control research of mountain-cultivated ginseng.Research on the chemical constituents and fingerprints of ginseng dispensing granules provided the scientific basis for the improvement of the quality of ginseng dispensing granules.