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OBJECTIVE To provide reference for quality control of Gentiana rhodantha. METHODS Taking 52 batches of G. rhodantha as subject， ultra-high performance liquid chromatography （UPLC） fingerprint was adopted. The similarity of 52 batches of medicinal materials samples was evaluated by the Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine （2004A edition）； the content of mangiferin was determined； chemometric analyses [cluster analysis， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）] were performed. RESULTS UPLC fingerprints of 52 batches of G. rhodantha were established， 17 common peaks were identified， and 6 of them were identified， which were loganic acid （peak 1）， neomangiferin （peak 3）， swertiamarin （peak 5）， dangyin （peak 6）， mangiferin （peak 7） and isoorientin （peak 9）. The similarities of 52 batches of medicinal materials samples were all greater than 0.9； cluster analysis showed that S1-S46， S48-S52 clustered into one class， and S47 alone； PCA results showed that the cumulative variance contribution rate of the first six principal components was 82.928%； OPLS-DA results showed that the corresponding components of swertiamarin， mangiferin and chemical composition represented by peak 4， 14， 15， 16 were the main iconic components affecting the quality differences of G. rhodantha medicinal materials. The contents of mangiferin in 52 batches of medicinal material samples ranged from 18.2 to 101.0 mg/g， mostly in accordance with 2020 edition of Chinese Pharmacopoeia. CONCLUSIONS The established UPLC fingerprint and chemometric analysis methods combined with content determination method of mangiferin can comprehensively evaluate the quality of G. rhodantha.
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OBJECTIVE: To apply DNA barcoding coupled with high resolution melting analysis to distinguish Gentiana rhodantha from its adulterants. METHODS: The internal transcribed spacer 2 (ITS2) barcode was selected for HRM analysis to produce standard melting profile of the selected species. RESULTS: The ITS2 molecular regions coupled with HRM analysis can effectively differentiate five herbal species, including two Gentiana rhodantha and their four common adulterants. CONCLUSION: DNA barcoding coupled with HRM analysis is a accurate, reliable, rapid, cost-effective and robust tool, which may contribute to the quality control of traditional Chinese medicine in the natural health product industry.
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Objective To explore the bioactive constituents of Gentiana rhodantha for its serum pharmacochemistry research.Methods The blood migrating constituents of Gentiana rhodantha were determined by comparing the HPLC fingerprints of the aqueous extracts,drug contained serum and blank serum.Results Twenty compounds were detected in drug contained serum,four among which were original constituents of Gentiana rhodantha,the rest might be metabolites of the original ingredients.Conclusion These twenty constituents absorbed in blood could be the bioactive components of Gentiana rhodantha.Further studies will be useful to clarify the bioactive constituents and action mechanisms of Gentiana rhodantha.
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OBJECTIVE: To predict the potential geographical distribution of Gentiana rhodantha and study the relationship between species distribution, chemical compounds content and ecological factors in Yunnan-Guizhou Plateau. METHODS: MaxEnt modeling combined with geographic information system (GIS) was used to predict the potential geographical distribution of G. rhodantha. Ultra performance liquid chromatography (UPLC)was used to establish the UPLC fingerprints of different parts of medicinal materials. Pearson correlation analysis and stepwise regression analysis were used to analyze the relationship between various chemical compounds and ecological factors. RESULTS: The AUC of the ROC curves of the training data and test data were 0.919 and 0.915, respectively, which indicated that the predictive results with the maximum model were highly precise and exact. The results of species distribution modeling showed that the main environmental factors determining the potential distribution were annual temperature range (the most suitable range: 16.0-27.0℃), mean diurnal range (the most suitable range: 8.5-11.6℃), average monthly precipitation of June (the most suitable range: 200-400 mm), average monthly precipitation of September (the most suitable range: 90-125 mm), average monthly maximum temperature of June (the most suitable range: 21.0-27.0℃), temperature seasonality (the most suitable range: 4 000-5 200), average monthly precipitation of October (the most suitable range: 65-110℃), average monthly minimum temperature of July (the most suitable range: 14.5-20.5℃), average monthly maximum temperature of January (the most suitable range: 14.5-25.5℃), subsoil pH (the most suitable range: pH<5), and average monthly precipitation of April (the most suitable range: 14.5-25.5 mm). Correlation analysis showed that the contents of mangiferin, total compounds and their ratios of aerial part and underground part had significant relationship with temperature, precipitation and hysico-chemical properties of top soil (P<0.05 or P<0.01) and had significant negative (P<0.05) or most significant negative relationship with habitat suitability (P<0.01). The variation of temperature and precipitation of June to August and subsoil pH subsoil CEC were the key ecological factors of accumulation of the chemical compounds in G.rhodantha. CONCLUSION: The best growing areas for G. rhodantha are mainly located in Middle and West Yunnan, Southeast and South Guizhou.
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Objective: To develope a method for UV-Vis and UPLC fingerprint on various medicinal parts of Gentiana rhodantha. The chemical compounds variation in roots, steams, leaves, and flowers were studied by using fingerprint data combined with multivariate analysis. Methods: Using Shim-pack XR-ODS III liquid chromatographic column (150 mm × 2.0 mm, 2.2 μm) for gradient elution, mobile phase was water with 0.1% formic acid and acetonitrile, temperature was 40℃, detection wavelength was 242 nm, injection value was 0.3 μL, and flow rate was 1.00 mL/min. Electrospray ionization (ESI) source and MRM model, the interface voltage was set to 3.5 kV. Desolation line (DL) temperature was 250℃. Nebulizing gas and drying gas were nitrogen at a flow rate of 3.0 and 15.0 L/min, respectively. Collision gas was 0.15 mL/min. UV-Vis detection wavelength range at 200 - 500 nm, slit was 1.0 nm, step was 0.5 nm. Multivariate analysis methods including partial least squares discriminant analysis (PLS-DA), variable importance (VIP), and hierarchical cluster were used for this projection. Results: The investigation of UPLC and UV-Vis showed RSD was lower than 2.00% for precision, repeatability, and stability. The recoveries of loganin acid, mangiferin, and sweroside were 97.89% - 102.71% and RSD was 1.09% - 2.88%. Pretreatment by 11 smooth + first order was the best data processing method for PLS-DA model (R2cal = 0.9315, RMSEE = 0.302, R2val = 0.901, and RMSEP = 0.341). UV-Vis spectra of roots, steams, leaves, and flowers had the significant fingerprint characteristic. Similarity analysis showed the chemical compounds in the leaves and flowers were similar and those in the stems and roots were similar too. Similarity index of root medicinal material has a widely range. The quality of the roots was not stable. The contents of loganic acid and mangiferin were the highest in the leaves and flowers [(1.46 ± 0.42) and (51.59 ± 15.45) mg/g]. The content of sweroside was the hightest in the roots [(4.41 ± 3.24) mg/g]. The total content of compounds of the leaves was higher than other medicinal part, the three compounds, loganic acid, mangiferin, and sweroside were very used for the discrimination of different parts of G. rhodantha. Cluster anslysis showed there were geography characteristics of the constituents. Sample collected from high altitude was higher than samples collected from lower altitude. And they were classified in different groups. Conclusion: UPLC and UV-Vis fingerprint could describe the variation of chemical compounds in the roots, steams, leaves, and flowers. All the results could provide the evaluation method and basic theory of G. rhodantha resource.