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Objective: To study the relationship between the quantitative color value of the powder and the known component content of rhubarb charcoal, and lay the foundation for the establishment of the rhubarb charcoal processing process control and endpoint judgment based on the color quantitative value. Methods： Rhubarb charcoal samples were prepared at different temperatures and time. Based on the empirical judgment of the rhubarb charcoal processing, the visual analyzer and UV-Vis were used to quantify the color of the pieces and powder of rhubarb charcoal under different processing conditions. At the same time, the HPLC fingerprint method was used to evaluate the dynamic changes of chemical components during the processing of rhubarb charcoal, and the quantitative value of the color of the sample during the processing of rhubarb charcoal was correlated with the characteristic components of the HPLC fingerprint using the multivariate statistical method. Results: During the processing of rhubarb charcoal, as the degree of carbonization increased, the apparent color of the sample changed from light yellowish brown to burnt black. There was a high correlation between the lightness value (L*), red-green value (a*) of the sample pieces and powder and the yellow and blue values (b*). The area of the 26 characteristic peaks had varying degrees of correlation with the chromaticity value. Among the 14 known components, five bound anthraquinones (aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, chrysophanol-8-O-β-D- glucoside, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside), and two sennosides (sennoside A, sennoside B) had a linear positive correlation with the chromaticity value. The content of five free anthraquinones (aloe-emodin, rhein, chrysophanol, emodin, and physcion), gallic acid and 5-hydroxymethylfurfural (5-HMF), whose contents increased first and then decreased, showed a quadratic correlation with the chromaticity value. Conclusion: The subjective judgment of rhubarb charcoal in the processing process is consistent with the quantitative color value analysis. The quantitative color value has a clear correlation with the content of 14 active chemical components. It is preliminarily inferred that the color quantitative value can be used as the quality of the rhubarb charcoal processing process control and end-point determination indicators to achieve efficient and rapid identification of the processing quality of rhubarb charcoal, which can provide new ideas for the monitoring and quality control of the rhubarb charcoal processing process.
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Objective: To investigate the anti-hepatoma active components of Rhei Radix et Rhizoma and their molecular mechanisms through GEO database, integrative pharmacology platform and molecular docking technology. Method: The active ingredients of Rhei Radix et Rhizoma were screened by TCMIP and the corresponding targets of these components were predicted through TCMIP and Swisstarget databases. The hepatoma gene chip database was downloaded from GEO databases, and the differentially expressed genes between hepatocellular carcinoma (HCC) and normal liver tissue were analyzed by GEO2R. Based on the matching results of potential targets of Rhei Radix et Rhizoma and the targets of hepatoma, the key targets of Rhei Radix et Rhizoma against hepatoma were screened, and GO function enrichment and KEGG pathway enrichment analysis of the key targets were performed. Main components and core targets of Rhei Radix et Rhizoma against hepatoma were analyzed and screened by constructing PPI network, component-target network and traditional Chinese medicine-component-target-pathway network. Furthermore, the molecular docking between the core targets and the main active components was performed by Schrodinger-Maestro software to virtually verify their binding ability and analyze their binding mode. Result: A total of 20 anti-hepatoma active components of Rhei Radix et Rhizoma were collected and related 86 targets were obtained, including CDK1, AKR1C3, PTGS2, AR and CCNB1, etc. The results of GO functional enrichment mainly focused on the cell cycle, G2/M transition of mitotic cell cycle, oxidation-reduction process, drug reactions and steroid metabolism processes, etc. The results of KEGG pathway enrichment mainly involved cell cycle, cell senescence, complement system, arachidonic acid metabolism and bile metabolism, and these metabolic pathways were related to cell apoptosis, metastasis, inflammation and immunity. The results of molecular docking showed that 92.2% of the active components had good binding ability with the 10 core proteins, and the main combination forms mainly were hydrogen bonds, hydrophobic bonds, π-π bonds and cation-π. Conclusion: The active components of Rhei Radix et Rhizoma including rhein, emodin, chrysophanol-8-O-β-D-glucopyranoside, chrysophanol-1-O-β-D-glucoside and rhapontigenin can act on multiple targets such as CDK1, CCNB1, CYP2C9, MMP9 and PTGS2, by regulating signaling pathways related to cell apoptosis, metastasis, inflammation and immunity to play an anti-hepatoma effect.
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Objective: To establish a method for the determination of six components from Rumex chalepensis Mill.. Methods: The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion were simultaneously determined by HPLC. The mobile phase was methanol-0.1% formic acid, gradient elution, flow rate of 1 mL/min, column temperature of 25 ℃, injection volume of 5 μL, detected by Agilent Extend-C18 (250 mm × 4.6 mm, 5 μm) and diode array detector at 254 nm wavelength. Results: The content of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcionhad good linear relationship in the ranges of 208—3 120, 22.40—336.35, 178.9—2 908.8, 16.7—250.8, 104.4—1 566.0, 45.2—677.7 ng, respectively. The average recovery rates were 97.66%, 97.10%, 98.78%, 97.38%, 102.48%, and 95.51% (n = 6). The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion in 16 batches of R. chalepensis. were determined in the range of 0.6—7.1, 0—2.7, 1.0—6.5, 0.1—0.6, 0.7—4.3, and 0.1—0.4 mg/g, respectively. Sample contents of different growing years, harvesting dates, and plots were compared and analyzed. Two-year-old R. chalepensis. was collected in early spring or late summer and early autumn. The total content of six components was 12.2 mg/g, which was relatively high. Conclusion: The established method can be used for simultaneous determination of six components from R. chalepensis, and determine the harvesting time and season of R. chalepensis, which provides a scientific basis for the formulation of quality evaluation criteria of R. chalepensis.
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Objective To evaluate the ability of the antiplatelet aggregation of ten anthraquinone derivatives in Rhei Radix et Rhizoma by using bioassay method, and to screen for the indicators that can be used to control the quality of rhubarb wine processing product. Methods Platelet aggregation instrument was used to determine the platelet aggregation rate induced by ADP in vitro and calculate the antiplatelet aggregation rates of 10 anthraquinone derivatives (aloe-emodin, rhein, emodin, chrysophanol, physcion, aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside, chrysophanol-8-O-β-D-glucoside, and physcion-8-O-β-D-glucoside) at different concentrations. The biopotency was calculated by bioavailability software. In order to verify the accuracy of the bioassay results, the estimated inhibition constant of rhein and chrysophanol-8-O-β-D-glucoside in P2Y12 protein receptor were determined by molecular docking software. Results The bioassay results showed that the antiplatelet potencies of rhein and emodin were significantly higher than those of aloe-emodin, chrysophanol and physcion. Compared with the antiplatelet biopotency of aspirin, the antiplatelet potencies of rhein and emodin were 5.02 and 5.15 times higher than that of aspirin, which indicated that rhein and emodin have strong inhibitory effect on ADP-induced platelet aggregation. However, the antiplatelet potencies of aloe-emodin, chrysophanol and physcion were equivalent of that of aspirin. The antiplatelet potencies of aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside, chrysophanol-8-O-β-D-glucoside, and physcion-8-O-β-D-glucoside were higher 4.13, 4.46, 9.31, 5.46, and 7.80 times than that of aspirin, respectively, which indicated that the five anthraquinone glucosides had a strong ability to antagonize ADP-induced platelet aggregation. The results of molecular docking showed that P2Y12 protein had different selectivity to 10 anthraquinone derivatives, especially for rhein, chrysophanol-8-O-β-D-glucoside, and the estimated Ki value were 5.73 and 2.51 μmol/L, respectively, indicating that rhein, chrysophanol-8-O-β-D-glucoside produced a strong inhibitory effect on P2Y12 protein at a lower concentration level. Moreover, the activity of chrysophanol-8-O-β-D-glucoside was stronger than rhein, which was consistent with their measured intensity of antiplatelet aggregation. Conclusion All results showed that there were some differences among antiplatelet potencies of 10 anthraquinone derivatives, and finally the screening of rhein, emodin can be used as evaluation indicators to control the quality of rhubarb wine processing product.