likviriton and baicalin

Liang-Ge-San (LGS), a traditional Chinese medicine (TCM) formula, is usually used in acute inflammatory diseases in China.. This study aims to detect the optimal combination of anti-inflammatory components from LGS.. Four mainly representative components (phillyrin, emodin, baicalin, and liquiritin) from LGS were chosen. The optimal combination was investigated by orthogonal design study. Zebrafish inflammation model was established by lipopolysaccharide (LPS)-yolk microinjection, and then the anti-inflammatory activities of different combinations were determined by survival analysis, changes on inflammatory cells infiltration, the MyD88/NF-κB and MAPK pathways and inflammatory cytokines production.. The different combinations of bioactive ingredients from LGS significantly protected zebrafish from LPS-induced inflammation, as evidenced by decreased recruitment of macrophages and neutrophils, inhibition of the MyD88/NF-κB and MAPK pathways and down-regulation of TNF-α and IL-6. Among them, the combination group 8 most significantly protected against LPS. The combination of group 8 is: 0.1 μM of emodin, 2 μM of baicalin, 20 μM of phillyrin and 12.5 μM of liquiritin.. The optimized combination group 8 exerts the most significant anti-inflammatory activity by inhibiting the recruitment of inflammatory cells, activation of the MyD88/NF-κB and MAPK pathways and the secretion of pro-inflammatory cytokines. This present study provides pharmacological evidences for the further development of new modern Chinese drug from LGS to treat acute inflammatory diseases, but indicated the use of zebrafish in the screening of components from formulas.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Drugs, Chinese Herbal; Emodin; Flavanones; Flavonoids; Glucosides; Inflammation; Interleukin-6; Larva; Lipopolysaccharides; Macrophages; MAP Kinase Signaling System; Medicine, Chinese Traditional; Myeloid Differentiation Factor 88; Neutrophil Infiltration; Neutrophils; NF-kappa B; Tumor Necrosis Factor-alpha; Yolk Sac; Zebrafish; Zebrafish Proteins

A simple and accurate high-performance liquid chromatography-photodiode array (HPLC-PDA) detection method has been developed and validated for simultaneous determination of nine components-liquiritin, coptisine, baicalin, palmatine, berberine, wogonoside, baicalein, glycyrrhizin and wogonin-in the traditional Korean formula, Banhasasim-tang decoction. A Gemini C18 analytical column was used to separate the nine constituents and kept at 40°C by gradient elution with 0.1% (v/v) trifluoroacetic acid in distilled water (A) and acetonitrile (B) as mobile phases. The flow rate was 1.0 mL/min and the injection volume was 10 µL. The PDA detection wavelengths were set at 254, 275 and 350 nm. Calibration curves of all compounds showed good linearity with coefficients of determination ≥0.9998 within the test ranges. The limits of detection and quantification of all compounds were in the range 0.01-0.09 and 0.03-0.30 µg/mL, respectively. All recoveries of the nine marker compounds ranged from 98.65 to 103.22% with relative standard deviation (RSD) values <1.25%. The RSDs of intraday and interday precision were <1.13 and 1.83%, respectively. The concentrations of the nine marker constituents were 0.19-41.09 mg/g.

Topics: Acetonitriles; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Glucosides; Glycyrrhizic Acid; Humans; Limit of Detection; Medicine, Korean Traditional; Observer Variation; Pinellia; Plants, Medicinal; Reproducibility of Results; Solvents; Trifluoroacetic Acid; Water

To analyse and compare the characteristics of the intestinal absorption of puerarin, baicalin, berberine and liquiritin in different combinations of Gegenqinlian decoction based on pharmacokinetic parameters, a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was applied for the quantification of four components in rat's plasma. And pharmacokinetic parameters were determined from the plasma concentration-time data with the DAS software package. The influence of different combinations on pharmacokinetics of four components was studied to analyse and compare the absorption difference of four components, together with the results of the in vitro everted gut model and the rat single pass intestinal perfusion model. The results showed that compared with other combinations, the AUC values of puerarin, baicalin and berberine were increased significantly in Gegenqinlian decoction group, while the AUC value of liquiritin was reduced. Moreover, the absorption of four components was increased significantly supported by the results from the in vitro everted gut model and the rat single pass intestinal perfusion model, which indicated that the Gegenqinlian decoction may promote the absorption of four components and accelerate the metabolism of liquiritin by the cytochrome P450.

Topics: Animals; Area Under Curve; Berberine; Chromatography, High Pressure Liquid; Coptis; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhiza uralensis; Intestinal Absorption; Isoflavones; Male; Plant Roots; Plants, Medicinal; Pueraria; Random Allocation; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Tandem Mass Spectrometry

To study the intestinal dynamics characteristics of puerarin and other active ingredients in Gegenqinlian decoction.. Caco-2 cell monolayer model was used to research the transport of puerarin and baicalin, berberine hydrochloride, liquiritin when they were used in different compatibilities, meanwhile a ultra performance liquid chromato-graphic method was also developed.. The concentration of puerarin, haicalin, berberine hydrochloride, liqpiritin in cell culture medium was measured and the apparent permeahility coefficients (Papp) as well as percent absorption (%) from apical side to basolateral side were calculated. The absorption characteristics of the four components were different when they were used in different compatibilities. Furthermore the Papp sequence of puerarin in specific groups were puerarin-baicalin-berberine-liquiritin group > puerarin-baicalin group > puerarin-berberine > puerarin-liquiritin group > single puerarin (P < 0. 05).. The intestinal absorption dynamics process of the active ingredients in Gegenqinlian decoction changes as the change of prescription compatibilities. And the intestinal absorption characteristics of puerarin is enhanced by the use of other components.

Topics: Berberine; Biological Transport; Caco-2 Cells; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Humans; Intestinal Absorption; Intestinal Mucosa; Isoflavones; Medicine, Chinese Traditional; Plants, Medicinal

The aim is to study the intestinal absorption of different combinations of active compounds out of Gegenqinlian decoction. Rat single pass intestinal perfusion model with jugular vein cannulated was used. Samples were obtained continuously from the outlet perfusate and the mesenteric vein. The levels of puerarin, daidzin, liquilitin, baicalin, wogonoside, jatrorrhizine, berberine and palmatine were determined by LC-MS/MS and their permeability coefficients were calculated. The results showed that Glycyrrhiza could promote the absorption of the active ingredients in Pueraria which is the monarch herb; meanwhile, Pueraria also played a role in promoting the absorption of liquilitin. Based on the Gegenqinlian decoction and the different combinations experiments, the results concerning the absorption of baicalin and wogonoside were as follows. For baicalin, Pueraria and Glycyrrhiza could promote its absorption and the effect of Pueraria was more obvious. For wogonoside, Pueraria could also promote its absorption, while Glycyrrhiza played a opposite role. Pueraria and Glycyrrhiza both played a part in promoting the absorption of jateorhizine, berberine and palmatine, the effective compounds in Coptis.

Topics: Animals; Berberine; Berberine Alkaloids; Coptis; Drug Combinations; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhiza; Intestinal Absorption; Intestinal Mucosa; Isoflavones; Male; Plant Roots; Plants, Medicinal; Pueraria; Rats; Rats, Sprague-Dawley; Rhizome; Scutellaria baicalensis

A specific ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method was developed for the simultaneous determination of puerarin, daidzein, baicalin, wogonoside and liquiritin in rat plasma. Chromatographic separation was performed on a C(18) column packed with 1.7 microm particles by a linear gradient elution. The analytes and carbamazepine (internal standard, I.S.) were monitored in a selected-ion reaction (SIR) mode with a positive electrospray ionization (ESI) interface by the following ions: m/z 417.2 for puerarin, m/z 255.2 for daidzein, m/z 271.0 for baicalin, m/z 461.0 for wogonoside, m/z 441.0 for liquiritin and m/z 237.2 for carbamazepine (I.S.), respectively. The calibration curves of these analytes were linear over the concentration ranges from 0.00254-1.02 microg mL(-1) to 0.0102-10.2 microg mL(-1). Within-batch and between-batch precisions (RSD%) were all within 15% and accuracy (RE%) ranged from -10% to 10%. The extraction recoveries were on average 79.8% for puerarin, 90.8% for daidzein, 74.4% for baicalin, 70.2% for wogonoside and 84.7% for liquiritin. The validated method was successfully applied to investigate the pharmacokinetics of five bioactive compounds of GegenQinlian decoction (GQD) in rats.

Topics: Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Isoflavones; Mass Spectrometry; Rats; Sensitivity and Specificity