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This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.
Subject(s)
Animals , Mice , Biological Products , Chromatography, High Pressure Liquid , Coleoptera , Cyclooxygenase 2 , Diarrhea/drug therapy , Antineoplastic AgentsABSTRACT
OBJECTIVE： To study the difference of the contents of copper， lead， cadmium， mercury and arsenic in the single decoction and combined decoction of Huangqi jianzhong decoction （containing Astragali radix， Cinnamomi ramulus， Paeoniae radix alba， prepared Glycyrrhiza uralensis， Zingiberis rhizoma recens， Jujubae fructus）， and to provide reference for safe use of TCM in clinic. METHODS： Samples of medicinal materials， single decoction and combined decoction were prepared from different medicinal materials； the contents of copper， lead， cadmium， mercury and arsenic in various samples were determined by atomic absorption spectrophotometry， which were compared with the limits of copper （＜20 mg/kg）， lead （＜5 mg/kg）， cadmium （＜0.3 mg/kg）， mercury （＜0.2 mg/kg） and arsenic （＜2 mg/kg） content stated in 2015 edition of Chinese Pharmacopeia and Green Trade Standards of Importing Exporting Medicinal Plants Preparations. The difference of 5 elements contents were compared between single decoction and combined decoction. RESULTS： Among the 6 medicinal materials， the cadmium content of Cinnamomi ramulus and Zingiberis rhizoma recens exceeded the standard， 0.32， 0.74 mg/kg， respectively （copper， lead， mercury and arsenic all met the requirements）， and the 5 elements content of other medicinal materials all met the requirements； the contents of copper， lead， cadmium， mercury and arsenic in the single and combined decoction meet the requirements； compared with the medicinal materials， the contents of copper， lead， cadmium， mercury and arsenic in the single decoction were lower （P＜0.01）； compared with mixture of single decoction， the contents of copper and mercury in combined decoction were lower （P＜0.01）， while the contents of lead， cadmium and arsenic in the decoction were higher （P＜0.01）. CONCLUSIONS： For Huangqi jianzhong decoction， combined decoction can reduce the dissolution of copper and mercury， and increase the dissolution of lead， cadmium and arsenic. The possible reason is that the interaction of various components in the co-decoction process changes the solubility of copper， lead， cadmium， mercury and arsenic.
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To study the differences and similarities in pharmaceutical characterization and pharmacodynamic characterization between the single decoction and merger decoction of Baihu and Guizhi. The same technology parameters were used to prepare Baihu and Guizhi single decoticon and merger decoction extracts, and then the differences and similarities in pharmaceutical characterization were analyzed based on their HPLC fingerprint, content of index components, and the extraction content. The pharmacodynamic differences and similarities were analyzed by inflammatory model and pain model. There was no significant difference in HPLC chromatographic peak, but the peak area value reflected the difference of quantity to some extent. It was found that the peak value of single Rhizoma anemarrhenae and Cassia twig decoction was less than the peak of their merger decoction, but the peak value of single honey-fried Licorice root decoction was greater than the peak of merger decoction． The contents of neomangiferin, mangiferin and timosaponin B Ⅱ among index components as well as extraction content in merger decoction were higher than those in single decoction． The contents of liquiritin and glycyrrhizic acid as well as extraction content in merger decoction were lower than those in single decoction． There was no significant difference in the content of cinnamicacid and its extraction content between merger decoction and single decoction. According to the efficacy experiment, both of them showed significant anti-inflammatory and analgesic effects. However, the merger decoction showed faster anti-inflammation effect, and longer analgesic effect. It can be concluded that the merger decoction and single decoction of Baihu and Guizhi have the same material basis, and the merger decoction is better for the dissolution of the active ingredients in this recipe, and is more beneficial to the therapeutic effect.
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OBJECTIVE:To investigate the decoction amount changes of verbascoside,gallic acid in single decoction and com-bined decoction of rehmanniae radix preparata,corni fructus and poria,and provide scientific basis for further study of effective substance basis. METHODS:The single decoction,combined decoction with each 2 medicine and combined decoction with the 3 medicines of rehmanniae radix preparata,corni fructus and poria,were respectively prepared. HPLC was adopted to detect and com-pare the decoction amount of active component verbascoside in rehmanniae radix preparata and active component gallic acid in cor-ni fructus in each group. RESULTS:Compared with single decoction of rehmanniae radix preparata,the decoction amount of ver-bascoside was decreased in combined decoction of poria+rehmanniae radix preparata(P<0.01),decoction amount of verbascoside was increased in combined decoction of corni fructus+rehmanniae radix preparata or combined decoction with the 3 medicines(P<0.01). Compared with single decoction of corni fructus,the decoction amount of gallic acid in each combined decoction was de-creased (P<0.01). CONCLUSIONS:The combined decoction of rehmanniae radix preparata,corni fructus and poria has certain promotion and inhibition effects on the decoction of verbascoside in rehmanniae radix preparata and inhibition effect on the decoc-tion of gallic acid in corni fructus. It is speculated verbascoside may be one of the main components in combined decoction playing the role of effectiveness.
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Objective To compare the content variation of catalpol between single and mixed decocting Rehmanniae Radix and Paeoniae Radix alba; To preliminarily explore the variation rules of compatibility of Rehmanniae Radix and Paeoniae Radix alba. Methods HPLC method was used with Waters Symmetry C18 column (4.6 mm×150 mm, 5 μm), flow rate of 1.0 mL/min, detection wavelength of 210 nm, column temperature at 25 ℃. The mobile phase was acetonitrile-0.01% phosphoric acid in water isocratic elution system. Results Catalpol regression equation was Y=270 054X-57 724, r=0.999 6, showing good linear range among 0.225–7.2 μg. The average recovery rate was 101.15%, RSD=0.39%. The average content of catalpol in the mixed decoction of Rehmanniae Radix and Paeoniae Radix alba was 18.66%more than the single decoction of Rehmanniae Radix. Conclusion The content of catalpol in mixed decoction is higher than the single decoction of Rehmanniae Radix and Paeoniae Radix alba. The mixed decoction is favorable for the dissolving of catalpol from the medicine. The compatibility of Rehmanniae Radix and Paeoniae Radix alba is rational.
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Objective: To study the correlation of combinations in different proportions of Angelicae Sinensis Radix and Carthami Flos and explore the regularity in proportion of the two drugs compatibility. Methods: Data mining technology was applied to screening the proportions of Angelicae Sinensis Radix and Carthami Flos. The changes of the contents of ferulic acid, hydroxysafflor yellow A, chlorogenic acid, and coumaric acid in the single and mixed decoction of Angelicae Sinensis Radix and Carthami Flos were determined by HPLC. The experiments of thrombin time and scavenging DPPH were carried out to evaluate the pharmacological activities of compatibility between Angelicae Sinensis Radix and Carthami Flos. Results: The top 10 proportions of Angelicae Sinensis Radix and Carthami Flos were screening. The contents of ferulic acid, hydroxysafflor yellow A, chlorogenic acid, and coumaric acid were higher than the single decoction of Angelicae Sinensis Radix and Carthami Flos. Thrombin time was prolonged and the activity of scavenging DPPH was enhanced compared to the same drug dosage of the single decoction of Angelicae Sinensis Radix and Carthami Flos. Conclusion: Mixed decoction of Angelicae Sinensis Radix and Carthami Flos is rational and good for the stripping of aromatic acids from the drugs, which has some correlation with the change of activities of the compatibility between Angelicae Sinensis Radix and Carthami Flos.
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OBJECTIVE:To evaluate the effect of single decoction vs.mingled decoction on the content of Baicalin in Radix Scutellariae.METHOD:The separation of samples was performed on Agilent C18 column(250 mm?4.6 mm,5 ?m)with a column at room temperature.The mobile phase consisted of 0.5% formic acid-methano and 0.5% formic acid solution(55∶45)with flow rate of 1.0 mL?min-1 and detection wavelength of 274 nm.RESULTS:The linear range of Baicalin was 0.258 5～2.326 5 ?g(r=0.999 9)with an average recovery of 97.8%(RSD=1.1%,n=9).CONCLUSION:The content of Baicalin in Radix Scutellariae decocted alone is higher than in the mixture of all the single decoctions,and significantly higher than in Radix Scutellariae decocted by mingled decoction,all showing significant differences.
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OBJECTIVE: To explore the inner quality of two kinds of single decoction solution. METHODS: The methods of HPLC, UV, TLC etc. were used for the qualitative and quantitative analysis. RESULTS: The contents of main components in single decoction solution of herbs in granule form were higher than those in slice form- CONCLUSION: The decocting rate of herbs in granule form is higher. It is worth popularizing.