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The anti-inflammatory activity and compatibility ratio of flavonoids in Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle(GR) and Angelicae Sinensis Radix(AS) were evaluated by the superoxide anion scavenging test. The matrix formula of gel was optimized by orthogonal test design and the model of deep partial-thickness scald in mice was induced. The gel was applied to the wound. The tissue water content, wound healing rate, serum TNF-α and IL-1, and EGF and VEGF in tissues were measured at diffe-rent periods. The results revealed that when the compatibility ratio of GR and AS was 1∶2, the maximal scavenging efficacy on supe-roxide anion was observed. The gel displayed the optimal properties when carbomer(1%), glycerol(5%), propylene glycol(10%) were added into the matrix. Gel external application can significantly improve the wound healing rate, relieve tissue edema, diminish tissue water content, alleviate inflammatory reaction, and increase the content of EGF and VEGF in tissues(P<0.05). The gel prepared in the present study is effective in promoting granulation, relieving pain, resisting inflammation, and alleviating edema, and is potent in healing scalds.
Subject(s)
Animals , Mice , Anti-Inflammatory Agents , Drugs, Chinese Herbal , Flavonoids , Glycyrrhiza , RhizomeABSTRACT
Objective: Based on response surface methodology, HPLC was applied to quantitatively determine the optimal processing technology of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (GRRPM) from the perspective of multi-index and comprehensive evaluation. Methods: HPLC was used for quantitative analysis, and the content of liquiritin, liquiritigenin, licochalcone A and glycyrrhetinic acid was used as inspection indexes. Response surface methodology was used to investigate the effects of the adding amount of honey, steaming and soaking time, frying temperature and frying time on the processing technology of GRRPM, and to optimize the optimal processing technology of GRRPM. Results: The chromatographic column was Diamonsil C18 (2) (4.6 mm × 200 mm, 5 μm); mobile phase was acetonitrile-0.1% phosphate aqueous solution, gradient eluting: 0-20 min, 12%-32% acetonitrile; 20-45 min, 32%-70% acetonitrile; 45-75 min, 70%-97% acetonitrile, with detection wavelength of 260 nm, column temperature of 20 ℃, and flow rate of 1 mL/min; Using liquiritin as internal standard, the relative correction factors of glycyrrhizin, licochalcone A, glycyrrhizinic acid and their relative correction factors were determined and calculated to be 0.56, 0.64 and 1.42, respectively. The optimum processing process of GRRPM was as follows: the amount of honey was 1/4, the soaking time was 15 min, frying pan bottom temperature was 160 ℃, and frying time was 13 min. Conclusion: The results of systematic adaptability investigation of the experimental content determination method meet the requirements. The best processing scheme of GRRPM optimized by response surface methodology is feasible and provides scientific basis for formulating quality standards and modern research of GRRPM.
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Objective: To screen the extraction process of Wuzi Yanzong plus and minus Recipe (WYR) by pharmacodynamics and long-term toxicity test, and optimize it by orthogonal experiment to determine the best extraction process of WYR. Methods Long-term toxicity experiments were carried out on different processed WYR samples. The penile erection latency and sex hormone levels in rats with kidney-yang deficiency were used as pharmacodynamic indicators. The results of long-term toxicity and efficacy experiments were combined to screen out the optimal process plan. Using the content of ingredients and the rate of ointment as indicators, the orthogonal test was used to screen the optimal level of different extraction processes and verified. Results Long-term toxicity test results showed that all mice survived healthily and no obvious toxicity was observed. The results of pharmacodynamic experiments showed that the extracts of Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were refluxed with 70% ethanol, and the remaining medicinal materials were extracted with 70% ethanol as the solvent to obtain the best extraction method of WYR. The suitable extraction process was as follows: Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were extracted three times with 10 times 70% ethanol for 2.0 h each time; The remaining medicinal materials were percolated with 10 times 70% ethanol at 1.5 mL/min. The medicinal herbs were firstly soaked for 16 h before percolation. Conclusion WYR can significantly improve the penile erection latency and sex hormone levels in rats with kidney yang deficiency. The optimal process conditions are reasonable and feasible, which provides a basis for the follow-up development of WYR.
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Objective: To establish the quality control method for high-quality Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle (GRRPM) in Xinshenghua Granules (XG). Methods: The HPLC fingerprint analysis method for high-quality GRRPM was developed. The method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining the six active constituents (liquiritin, isoliquiritin, glycyrrhizic acid, liquiritigenin, liquiritin apioside, and isoliquiritin apioside) was developed and evaluated by comparison of the quantitative results with external standard method. Results: The fingerprints of GRRPM were established by HPLC from 30 batches. Fourteen peaks were acquired as common fingerprint peaks and seven peaks among them were identified with chemical reference. The relative retention time of common peaks and the peak area ratio of some common peaks were used to differentiate high-quality products from general products as indicators for fingerprint evaluation. With liquiritin and glycyrrhizic acid as internal standards, QAMS was developed and the mean relative correlation factors (RCFs) of isoliquiritin, liquiritigenin, liquiritin apioside, and isoliquiritin apioside were 0.502, 0.578, 0.252, and 0.257, respectively. The specifications of high-quality GRRPM were established for XG. Conclusion: These methods could be used for quality control of high-quality GRRPM of XG.
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Fangji Huangqi Tang is a traditional Chinese medicine formula composed of Stephaniae Tetrandrae Radix, Astragali membranaceus Radix, Atractylodis Macrocephale Rhizoma, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle. At present, the compounds extracted from the Chinese materia medica in Fangji Huangqi Decoction are reported to be alkaloids, flavonoids, saponins, saccharides and terpenoids. Fangji Huangqi Decoction was identified as one of the main formula used to treat chronic glomerular nephritis, nephrotic syndrome, renal edema, and cardiac edema in clinical application of traditional Chinese medicine. In this paper, the research progress in the chemical constituents and their pharmacologic actions of Fangji Huangqi Decoction are reviewed, providing the scientific basis for its modernization research and comprehensive utilization.
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Objective: To analyze the changes of chemical ingredients before and after compatibility of Aconiti Lateralis Radix Praeparata (ALRP) and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (GRRPM), and to explore the possible mechanism of toxicity attenuation of their compatibility. Methods: The chemical ingredients in the decoction of ALRP and the decoction of compatibility of ALRP and GRRPM were comparatively researched by HPLC-MS. Single decoction of ALRP and compound decoction of compatibility of ALRP and GRRPM were prepared, and their HPLC-MS fingerprints were respectively established and determined by Q-TOF/MS under the same condition. Results: Twenty ingredients and their structures were identified from single decoction of ALRP; 32 ingredients and their structures were indentified from the decoction of ALRP and GRRPM, among them 4 from GRRPM, 28 from ALRP. And the alkaloid categories and contents of ALRP were significantly different before and after the compatibility with GRRPM. Conclusion: The compatibility with GRRPM could change the alkaloid composition in ALRP, which provides the experimental evidences for the toxicity attenuation mechanism of compatibility of ALRP and GRRPM.
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Objective: To study the changing rules of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (GRRPM) with the active components alignment (ACA, liquiritin-liquiritigenin-glycyrrhizin acid-isoliquiritinin) of tonifying spleen and replenishing qi. Methods: Taking single herbs, their medicine pairs, or their medicine compounds as test sample, the sample solution was prepared by traditional water decoction and determined by HPLC. Results: The contents of liquiritin-liquiritigenin-glycyrrhizin acid-isoliquiritinin in GRRPM were (12.14±0.31), (3.25±0.19), (38.75±1.77), and (1.40±0.25) mg/mL; The ACA contents in GRRPM-Ginseng Radix et Rhizoma (GRR) were (2.87±0.04), (0.74±0.02), (6.42±0.18), and (0.37±0.03) mg/mL; The ACA contents in GRRPM-Atractylodis Macrocephalae Rhizoma (AMR) were (5.99±0.01), (2.16±0.09), (22.40±1.80), and (0.77±0.02) mg/mL; The ACA contents in Sijunzi Decotion (SD) were (2.99±0.03), (0.81±0.04), (10.05±0.31), and (0.32±0.03) mg/mL. Conclusion: Four main active components in the efficacy of the test components have almost the same change rule: GRRPM≤GRRPM-AMR≤SD≤GRRPM-GRR; The percentage change intervals of the ACA (liquiritin-liquiritigenin-glycyrrhizin acid-isoliquiritinin) are (7.76-9.34):(2.00-2.81):(17.35-31.41):1. The total contents in single herbs are three times as much as in compatibility components, so the Chinese materia medica (CMM) are often used in the type of formula, which indicates that the utilized space of ACA resources is larger, ACA has the obvious effect of correspondence, and the quality standard of CMM should be marked ACA and medicinal components of biological potency as indicators.
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Objective: To establish an LC-MS/MS method for determination of glycyrrhetinic acid (GA) in plasma after ig administration of Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle (Zhigancao) Decoction in rats in vivo. Methods: Treating plasma with ethyl acetate protein precipitation and taking reseveratrol was used as the internal standard (IS), the residues were analyzed with LC-MS/MS system (Agilent ORBAX SB-C18 RP column, 50 mm x 2.1 mm, 1.8 μm) with the mobile phase consisted of acetonitrile-water (80 : 20, 0.2% formic acid), flow-rate of 0.2 mL/min, and column temperature of 30 °C. Agilent 6410 triple quad mass spectrometer system with an electrospray ionization (ESI) source and negative ion mode was used as the detector in this study. Multi-reaction monitoring (MRM) was used to detect GA by using ion combinations of m/z 469.4→425.4 for GA and 227.1→143.0 for IS. Results: The calibration curve showed good linear regression (R2 = 0.997 1) within measurement ranges (33.4-8 560.0 ng/mL). Both the intra- and inter-day precision and variation RSD were less than 10%. The average recovery rates at low, medium, and high concentrations of GA were 75.3%, 78.2%, and 78.5%, respectively. Under these conditions, the pharmacokinetic parameters of GA in plasma of rats after ig administration of Zhigancao Decoction were t max (8.00 ± 1.13) h, Cmax (811.02 ± 300.25) ng/mL, and AUC0.24 (11 439.21 ± 3 367.36) ng/mL·h. Conclusion: The method provides a sensitive, accurate, precise, and reliable analytical procedure for detecting GA in rat plasma and studying its pharmacokinetics.