RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: As a traditional Chinese medicine (TCM) formula, Wu-tou decoction has been used for treating rheumatoid arthritis (RA) for more than a thousand years. Identifying pharmacodynamic constituents (PCs) of WTD and exploring their in vivo process are very meaningful for promoting the modernization of TCM. However, the pathological state might change this process. AIM OF THE STUDY: Hence, it is necessary and significant to compare the process in vivo of drugs both in normal and disease state and clarify their action mechanism. MATERIALS AND METHODS: Taking Wu-tou decoction (WTD) as the research object, a comprehensive strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to identify PCs, clarify and compare their absorption and distribution in normal and model rats, and then explore the potential mechanism of TCM. Firstly, the PCs in WTD were identified. Then, the pharmacokinetics (PK) and tissue distribution of these ingredients were studied. Finally, the constituents with the difference between normal and model rats were selected for target network pharmacological analysis to clarify the mechanism. RESULTS: A total of 27 PCs of WTD were identified. The absorption and distribution of 20 PCs were successfully analyzed. In the disease state, the absorption and distribution of all these components were improved to have better treatment effects. The results of target network pharmacological analysis indicated that PTGS1, PTGS2, ABCB1, SLC6A4, CHRM2, ESR1, ESR2, CDK2, TNF and IL-6 are 10 key targets for WTD against RA. The regulatory effects of WTD on the expression of PTGS2 and TNF were further verified. Pathway enrichment analysis showed that the key mechanism of WTD against RA is to reduce inflammation and regulate the immune response. CONCLUSION: These results indicated that this strategy could better understand the in vivo process and mechanism of WTD under the pathological state. Furthermore, this strategy is also appropriate for other TCM.
Asunto(s)
Antirreumáticos/química , Antirreumáticos/farmacología , Artritis Experimental/tratamiento farmacológico , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Administración Oral , Animales , Antirreumáticos/administración & dosificación , Antirreumáticos/farmacocinética , Artritis Experimental/inducido químicamente , Cromatografía Líquida de Alta Presión , Ciclooxigenasa 2/metabolismo , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/química , Inflamación/metabolismo , Lipopolisacáridos/toxicidad , Masculino , Espectrometría de Masas , Medicina Tradicional China , Redes y Vías Metabólicas/efectos de los fármacos , Ratones , Células RAW 264.7 , Ratas Sprague-Dawley , Distribución Tisular , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The aim of this study was to investigate the pharmacokinetics and pharmacodynamics of seven main active components of Mahuang decoction (MHD) and its time-concentration-effect relationship. The asthmatic rat model was established by the method of ovalbumin (OVA) sensttization. The plasma concentrations of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamic acid, glycyrrhizic acid in asthmatic model rat were investigated by a selective and rapid HPLC/MS-MS method. Simultaneously, the asthma-involved cytokines including leukotrienes B4 (LTB4), thromboxane B2 (TXB2), 6-Keto-Prostaglandin F1α (6-K-PGF1α) and histamine (HIS) levels in rat plasma were determined by using ELISA. A mathematics method was applied to assess the trend of percentage rate of change among different time intervals of the seven components. The sigmoid E max function was used to establish the PK-PD modeling of MHD. The results indicated that MHD could control or ameliorate asthma. There was a hysteresis between the peaked drug concentration and maximum therapeutic effect of MHD. The PK-PD curves of MHD showed clockwise or counter-clockwise hysteresis loop. In addition, amygdalin might exert a more significant influence on regulating cytokines levels in asthmatic rats among the seven components of MHD.
Asunto(s)
Asma/tratamiento farmacológico , Preparaciones de Plantas/farmacología , Preparaciones de Plantas/farmacocinética , Amigdalina/sangre , Animales , Asma/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Cinamatos/sangre , Correlación de Datos , Medicamentos Herbarios Chinos/farmacocinética , Medicamentos Herbarios Chinos/farmacología , Ephedra sinica , Efedrina/análogos & derivados , Efedrina/sangre , Flavanonas/sangre , Glucósidos/sangre , Ácido Glicirrínico/sangre , Masculino , Ovalbúmina/sangre , Ratas , Ratas Sprague-DawleyRESUMEN
Liquorice [main ingredient, glycyrrhizin (GL)] is widely used as a food sweetener and herbal medicine. Occasionally, liquorice consumption causes pseudoaldosteronism as a side effect which causes oedema, hypokalaemia, and hypertension due to hyperactivity of mineral corticoid receptor. We aimed to detect GL metabolites in human blood and urine samples and to determine the pathological relationship between GL metabolites and pseudoaldosteronism. For this multi-centre, retrospective, cross-sectional study, we recruited patients who had visited Center for Kampo Medicine in Keio University Hospital, Department of Japanese Oriental (Kampo) Medicine in Chiba University Hospital, Clinic of Japanese Oriental (Kampo) Medicine in Kanazawa University Hospital, and Department of Oriental Medicine in Kameda Medical Center from November 2011 to July 2018. We collected laboratory data including concentration of serum potassium, plasma activity of renin and aldosterone, and residual blood and/or urine samples of participants who had experienced symptoms/signs of pseudoaldosteronism in the form of increase in blood pressure and occurrence or aggregation of oedema while taking liquorice-containing herbal preparations, and measured GL metabolites using a highly selective liquid chromatography tandem mass spectrometer system. We registered 97 participants (mean age 60 ± 15 years; male:female 14:83). 18ß-glycyrrhetinic acid (GA) was detected in 67 serum samples (median 122 nM, range 5 nM-1.8 µM) and 18ß-glycyrrhetyl-3-O-sulfate (compound 3) in 68 samples (median 239 nM, range 2 nM-4.2 µM). 3-Monoglucuronyl 18ß-glycyrrhetinic acid, 22α-hydroxy-18ß-glycyrrhetyl-3-O-sulfate-30-glucuronide, 22α-hydroxy-18ß-glycyrrhetyl-3-O-sulfate, and GL itself were not or rarely detected. We could not find any correlation between blood pressure or peripheral oedema and serum concentration of GL metabolites. Sulfotransferase 2A1 catalysed the metabolic reaction of GA to compound 3, a major GL metabolite in human blood. High serum concentration of compound 3 was related to lower renin, aldosterone, and potassium levels, suggesting a pathological relationship between compound 3 and liquorice-induced pseudoaldosteronism. This is the first study to identify the association between a novel metabolite, compound 3, and the incidence of pseudoaldosteronism, highlighting it as a promising biomarker.
Asunto(s)
Glycyrrhiza/toxicidad , Ácido Glicirrínico/sangre , Síndrome de Liddle/inducido químicamente , Edulcorantes/toxicidad , Aldosterona/sangre , Biomarcadores/sangre , Estudios Transversales , Relación Dosis-Respuesta a Droga , Femenino , Glycyrrhiza/metabolismo , Ácido Glicirrínico/metabolismo , Humanos , Síndrome de Liddle/sangre , Síndrome de Liddle/metabolismo , Masculino , Persona de Mediana Edad , Potasio/sangre , Renina/sangre , Estudios Retrospectivos , Edulcorantes/metabolismoRESUMEN
A simple and sensitive liquid chromatography with tandem mass spectrometry method was developed for simultaneous quantification of paeoniflorin, albiflorin, oxypaeoniflorin, liquiritin, liquiritigenin, glycyrrhetinic acid, and glycyrrhizin in rat plasma after oral administration of Shaoyao-Gancao decoction, which is traditionally used in the treatment of polycystic ovary syndrome. The plasma samples were pretreated with methanol as precipitant. The method exhibited good linearity (correlation coefficient (R2 ) > 0.99) with lower quantification limits of 0.595-4.69 ng/mL for all analytes. Intra- and interbatch precision, accuracy, recovery, and stability of the method were all within accepted criteria. The results showed that the pharmacokinetic behaviors of the seven compounds were altered in the pathological status of polycystic ovary syndrome. Furthermore, a total of 36 metabolites were structurally identified based on their accurate masses and fragment ions. The major metabolic pathway involves phase I metabolic reactions (such as hydroxylation), phase II metabolic reactions (such as sulfation and glucuronidation conjugation) as well as the combined multiple-step metabolism. This study is the first report on the pharmacokinetic and metabolic information of Shaoyao-Gancao decoction in both normal and model rats, which would provide scientific evidences for the bioactive chemical basis of herbal medicines and also promote the clinical application of Shaoyao-Gancao decoction for treating polycystic ovary syndrome.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Administración Oral , Animales , Hidrocarburos Aromáticos con Puentes/sangre , Hidrocarburos Aromáticos con Puentes/metabolismo , Hidrocarburos Aromáticos con Puentes/farmacocinética , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/metabolismo , Medicamentos Herbarios Chinos/farmacocinética , Femenino , Flavanonas/sangre , Flavanonas/metabolismo , Flavanonas/farmacocinética , Glucósidos/sangre , Glucósidos/metabolismo , Glucósidos/farmacocinética , Ácido Glicirretínico/sangre , Ácido Glicirretínico/metabolismo , Ácido Glicirretínico/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/metabolismo , Ácido Glicirrínico/farmacocinética , Monoterpenos/sangre , Monoterpenos/metabolismo , Monoterpenos/farmacocinética , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en TándemRESUMEN
Glycyrrhizae Radix is widely used as herbal medicine and is effective against inflammation, various cancers, and digestive disorders. We aimed to develop a sensitive and simultaneous analytical method for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin, the four marker components of Glycyrrhizae Radix extract (GRE), in rat plasma using liquid chromatography-tandem mass spectrometry and to apply this analytical method to pharmacokinetic studies. Retention times for glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were 7.8 min, 4.1 min, 3.1 min, and 2.0 min, respectively, suggesting that the four analytes were well separated without any interfering peaks around the peak elution time. The lower limit of quantitation was 2 ng/mL for glycyrrhizin and 0.2 ng/mL for isoliquiritigenin, liquiritigenin, and liquiritin; the inter- and intra-day accuracy, precision, and stability were less than 15%. Plasma concentrations of glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were quantified for 24 h after a single oral administration of 1 g/kg GRE to four rats. Among the four components, plasma concentration of glycyrrhizin was the highest and exhibited a long half-life (23.1 ± 15.5 h). Interestingly, plasma concentrations of isoliquiritigenin and liquiritigenin were restored to the initial concentration at 4-10 h after the GRE administration, as evidenced by liquiritin biotransformation into isoliquiritigenin and liquiritigenin, catalyzed by fecal lysate and gut wall enzymes. In conclusion, our analytical method developed for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin could be successfully applied to investigate their pharmacokinetic properties in rats and would be useful for conducting further studies on the efficacy, toxicity, and biopharmaceutics of GREs and their marker components.
Asunto(s)
Chalconas/sangre , Flavanonas/sangre , Glucósidos/sangre , Ácido Glicirrínico/sangre , Administración Oral , Animales , Chalconas/farmacocinética , Cromatografía Liquida , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/farmacocinética , Flavanonas/farmacocinética , Glucósidos/farmacocinética , Ácido Glicirrínico/farmacocinética , Masculino , Extractos Vegetales/sangre , Extractos Vegetales/farmacocinética , Control de Calidad , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en TándemRESUMEN
Lizhong decoction (LZD), a classic formula, has been used to treat ulcerative colitis (UC) for thousands of years in clinical practice. However, the pharmacokinetic characteristics of its major bioactive components in rats under different physiological and pathological states are not clear. Thus, in this study, a rapid and sensitive analytical method, ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) method, was developed and applied to simultaneously determine glycyrrhizic acid, liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin, 6-gingerol, ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Re in normal and UC rats after oral administration of LZD extract. A Waters BEH C18 UPLC column was used for chromatographic separation, while acetonitrile and 0.1% formic acid were selected as mobile phase. The linearity of nine analytes was >0.9920. Inter- and intra-day accuracy was ≤ 11.4% and precision was from 1.1 to 12.7%. Additionally, stable and suitable extraction recoveries were also obtained. The established method was validated and found to be specific, accurate and precise for nine analytes. Furthermore, it was successfully applied to the pharmacokinetic investigation of nine major components after oral administration of LZD extracts to normal and model rats, respectively. The results showed that the pharmacokinetic parameters (Cmax , Tmax , AUC0-t , AUC0-∞ ) in the plasma of UC rats were significantly different from those of normal rats, which could provide a reference for the clinical application of LZD.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Colitis Ulcerosa/metabolismo , Medicamentos Herbarios Chinos/farmacocinética , Espectrometría de Masas en Tándem/métodos , Administración Oral , Animales , Chalcona/análogos & derivados , Chalcona/sangre , Chalcona/química , Chalcona/farmacocinética , Medicamentos Herbarios Chinos/administración & dosificación , Ginsenósidos/sangre , Ginsenósidos/química , Ginsenósidos/farmacocinética , Glucósidos/sangre , Glucósidos/química , Glucósidos/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/química , Ácido Glicirrínico/farmacocinética , Límite de Detección , Modelos Lineales , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los ResultadosRESUMEN
A highly sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of glycyrrhizin (GL) and its active metabolite, glycyrrhetinic acid (GA), from human plasma was validated and applied to a human pharmacokinetic study. The analytes were extracted from human plasma using an Oasis MAX cartridge and chromatographic separation was performed on an Inertsil ODS-3 column. The detection was performed using an API 4000 mass spectrometer operating in the positive electrospray ionization mode. Selected ion monitoring transitions of m/z 823 â 453 for GL and m/z 471 â 149 for GA were obtained. The response was a linear function of concentration over the ranges of 0.5-200 ng/mL for GL and 2-800 ng/mL for GA (both R2 > 0.998). Using this method, the pharmacokinetics of GL after single oral administration of a clinical dose (75 mg) to six healthy male Japanese volunteers were evaluated. GL was detected in the plasma of all subjects and the average peak concentration was 24.8 ± 12.0 ng/mL. In contrast, peak concentration of GA was 200.3 ± 60.3 ng/mL, i.e. ~8-fold higher than that of GL. This is the first report clarifying pharmacokinetic profiles of GL and GA simultaneously at a therapeutic oral dose of a GL preparation.
Asunto(s)
Cromatografía Liquida/métodos , Ácido Glicirretínico/sangre , Ácido Glicirretínico/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/farmacocinética , Administración Oral , Adulto , Ácido Glicirretínico/química , Ácido Glicirrínico/administración & dosificación , Ácido Glicirrínico/química , Humanos , Modelos Lineales , Masculino , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Masas en Tándem/métodosRESUMEN
Entecavir (ETV) is a superior nucleoside analogue used to treat hepatitis B virus (HBV) infection. Although its advantages over other agents include low viral resistance and the elicitation of a sharp decrease in HBV DNA, adverse effects such as hepatic steatosis, hepatic damage and lactic acidosis have also been reported. Glycyrrhizin has long been used as hepato-protective medicine. The clinical combination of ETV plus glycyrrhizin in China displays better therapeutic effects and lower rates of liver damage. However, there is little evidence explaining the probable synergistic mechanism that exists between these two drugs from a pharmacokinetics view. Here, alterations in the plasma pharmacokinetics, tissue distribution, subcellular distribution, and in vitro and in vivo antiviral activity of ETV after combination with glycyrrhizic acid (GL) were analysed to determine the synergistic mechanisms of these two drugs. Specific efflux transporter membrane vesicles were also used to elucidate their interactions. The primary active GL metabolite, glycyrrhetic acid (GA), did not affect the plasma pharmacokinetics of ETV but promoted its accumulation in hepatocytes, increasing its distribution in the cytoplasm and nucleus and augmenting the antiviral efficiency of ETV. These synergistic actions were primarily due to the inhibitory effect of GA on MRP4 and BCRP, which transport ETV out of hepatocytes. In conclusion, GA interacted with ETV at cellular and subcellular levels in the liver through MRP4 and BCRP inhibition, which enhanced the antiviral activity of ETV. Our results partially explain the synergistic mechanism of ETV and GL from a pharmacokinetics view, providing more data to support the use of these compounds together in clinical HBV treatment.
Asunto(s)
Antivirales/uso terapéutico , Ácido Glicirretínico/farmacología , Ácido Glicirrínico/farmacología , Guanina/análogos & derivados , Hepatitis B/tratamiento farmacológico , Animales , Antivirales/sangre , Antivirales/farmacocinética , Células Cultivadas , ADN Viral/sangre , Interacciones Farmacológicas , Ácido Glicirrínico/sangre , Ácido Glicirrínico/farmacocinética , Guanina/sangre , Guanina/farmacocinética , Guanina/uso terapéutico , Células Hep G2 , Hepatitis B/sangre , Virus de la Hepatitis B/genética , Hepatocitos/metabolismo , Humanos , Hígado/metabolismo , Masculino , Proteínas de Transporte de Membrana/genética , Ratones Transgénicos , Ratas Sprague-DawleyRESUMEN
Rapid, sensitive, selective and accurate UPLC-MS/MS method was developed and fully validated for simultaneous determination of cinnamaldehyde, cinnamic acid, 2-methoxy cinnamic acid, glycyrrhizic acid, glycyrrhetinic acid, liquiritigenin and isoliquiritin in rat plasma after oral administration of Guizhi-gancao decoction. Plasma samples were processed with a simple protein precipitation technique using acetonitrile, followed by chromatographic separation using a Thermo Hypersil GOLD C18 column. A 11.0min linear gradient elution was used at a flow rate of 0.2mL/min with a mobile phase of 0.1% acetic acid containing 0.2mM ammonium acetate in water and acetonitrile. The analytes and internal standard, schisandrin, were detected using both positive and negative ion electrospray ionization in multiple reaction monitoring mode. The developed method was validated for intra-day and inter-day accuracy and precision whose values fell in the acceptable limits. Matrix effect was found to be minimal. Recovery efficiency of all the analytes was found to be >60%. Stability results showed that the analytes were stable at all the conditions. This validated method was successfully used to study the pharmacokinetics of multiple compounds in rat plasma after oral administration of Guizhi-gancao decoction.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/farmacocinética , Plasma/química , Espectrometría de Masas en Tándem/métodos , Administración Oral , Animales , Chalcona/análogos & derivados , Chalcona/sangre , Chalcona/química , Cinamatos/sangre , Cinamatos/química , Medicamentos Herbarios Chinos/química , Flavanonas/sangre , Flavanonas/química , Glucósidos/sangre , Glucósidos/química , Ácido Glicirretínico/sangre , Ácido Glicirretínico/química , Ácido Glicirrínico/sangre , Ácido Glicirrínico/química , Masculino , Ratas , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: To investigate the effects of glycyrrhizic acid supplementation on glucose and lipid metabolism in rodents consuming a high-fat, high-sucrose diet. METHODS: Twenty-four male, 8-week old Sprague Dawley rats with an initial weight of 160 to 200 g were randomised into three groups (n = 6 for each group): groups A (standard rat chow), B (high-fat, high-sucrose diet), and C (high-fat, high-sucrose diet + 100 mg/kg/d of glycyrrhizic acid via oral administration). The rats were treated accordingly for 4 wk. Glycaemic parameters, lipid profile, stress hormones, and adiponectin levels were measured after the treatment. Relative gene expressions of peroxisome proliferator-activated receptor α and γ, lipoprotein lipase as well as gluconeogenic enzymatic activities in different tissues were also determined. RESULTS: Consumption of high-fat, high-sucrose diet triggered hyperglycaemia, insulin resistance, and dyslipidemia, which were effectively attenuated by supplementation with glycyrrhizic acid. Glycyrrhizic acid supplementation also effectively reduced circulating adrenaline, alleviated gluconeogenic enzymes overactivity, and promoted the upregulation of lipoprotein lipase expression in the cardiomyocytes and skeletal muscles. A high calorie diet also triggered hypoadiponectinaemia and suppression of peroxisome proliferator-activated receptor γ expression, which did not improve with glycyrrhizic acid treatment. CONCLUSION: Supplementation with glycyrrhizic acid could alleviate high calorie diet-induced glucose and lipid metabolic dysregulations by reducing circulatory stress hormones, normalizing gluconeogenic enzyme activities, and elevating muscular lipid uptake. The beneficial effects of these bioactivities outweighed the adverse effects caused by diet-induced repression of peroxisome proliferator-activated receptor γ expression, resulting in the maintenance of lipid and glucose homeostasis.
Asunto(s)
Antiinflamatorios/farmacología , Dieta Alta en Grasa/efectos adversos , Ácido Glicirrínico/farmacología , Síndrome Metabólico/prevención & control , PPAR gamma/antagonistas & inhibidores , Sacarosa/administración & dosificación , Adiponectina/sangre , Animales , Antiinflamatorios/sangre , Grasas de la Dieta/administración & dosificación , Epinefrina/sangre , Gluconeogénesis/efectos de los fármacos , Ácido Glicirrínico/sangre , Lipoproteína Lipasa/sangre , Lipoproteína Lipasa/efectos de los fármacos , Masculino , Síndrome Metabólico/sangre , PPAR gamma/sangre , Ratas , Ratas Sprague-DawleyRESUMEN
The present study was designed to investigate the effects of Laminaria japonica (Laminaria) on pharmacokinetics of glycyrrhetinic acid (GA) following oral administration of Liquorice extract in rats. Following oral administrations of single-dose and multi-dose Liquorice extract and Liquorice-Laminaria extract, respectively, plasma samples were obtained at various times and the concentrations of GA, liquiritigenin, and isoliquiritigenin were measured by LC-MS. The effects of Laminaria extract on pharmacokinetics of GA were also investigated, following single-dose and multidose of glycyrrhizic acid (GL). The effects of Laminaria extract on intestinal absorption of GA and GL were studied using the in situ single-pass intestinal perfusion model. The metabolism of GL to GA in the contents of small and large intestines was also studied. The results showed Liquorice-Laminaria extract markedly increased the plasma concentration of GA, accompanied by a shorter Tmax. Similar alteration was observed following multidose administration. However, pharmacokinetics of neither liquiritigenin nor isoliquiritigenin was affected by Laminaria. Similarly, Laminaria markedly increased concentration and decreased Tmax of GA following oral GL were observed. The data from the intestinal perfusion model showed that Laminaria markedly increased GL absorption in duodenum and jejunum, but did not affect the intestinal absorption of GA. It was found that Laminaria enhanced the metabolism of GL to GA in large intestine. In conclusion, Laminaria increased plasma exposures of GA following oral administration of liquorice or GL, which partly resulted from increased intestinal absorption of GL and metabolism of GL to GA in large intestine.
Asunto(s)
Interacciones Farmacológicas , Ácido Glicirretínico/sangre , Glycyrrhiza/química , Ácido Glicirrínico/farmacocinética , Absorción Intestinal , Laminaria , Extractos Vegetales/farmacología , Administración Oral , Animales , Ácido Glicirrínico/sangre , Mucosa Intestinal/metabolismo , Masculino , Extractos Vegetales/sangre , Extractos Vegetales/farmacocinética , Ratas Sprague-DawleyRESUMEN
Gan-Sui-Ban-Xia Decoction (GSBXD) was first presented by Zhang Zhongjing in the book Synopsis of Golden Chamber during the Han Dynasty period. The formula was then used for the treatment of persistent fluid retention with floating pulse in Traditional Chinese Medicine (TCM), which in modern medicine is known as malignant ascites. Here, a rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the determination of glycyrrhizinic acid, liquiritin, paeoniflorin, albiflorin after oral administration of GSBXD plus-minus Gansui and Gancao anti-drug combination to investigate the possible pharmacokinetic profile differences of different prescriptions with GSBXD in normal rats. The differences of pharmacokinetic parameters among groups were tested by the Student's t-test with p < 0.05 as the level of significance. Significant differences were found between the Gansui and Gancao anti-drug combination and other herbs in GSBXD on pharmacokinetic profile of glycyrrhizinic acid, liquiritin, paeoniflorin and albiflorin. The obtained knowledge might contribute to the rationality of the clinic use of GSBXD and also reveal the compatibility conditions of the Gansui and Gancao anti-drug combination.
Asunto(s)
Hidrocarburos Aromáticos con Puentes/farmacocinética , Medicamentos Herbarios Chinos/farmacocinética , Flavanonas/farmacocinética , Glucósidos/farmacocinética , Ácido Glicirrínico/farmacocinética , Monoterpenos/farmacocinética , Administración Oral , Animales , Ascitis/tratamiento farmacológico , Hidrocarburos Aromáticos con Puentes/sangre , Cromatografía Liquida , Combinación de Medicamentos , Euphorbia/metabolismo , Flavanonas/sangre , Glucósidos/sangre , Glycyrrhiza uralensis/metabolismo , Ácido Glicirrínico/sangre , Masculino , Medicina Tradicional China , Monoterpenos/sangre , Paeonia/metabolismo , Pinellia/metabolismo , Ratas , Ratas Wistar , Espectrometría de Masas en TándemRESUMEN
An LC-MS/MS method was developed for simultaneous analysis of puerarin, daidzin, baicalin, glycyrrhizic acid, liquiritin, berberine, palmatine and jateorhizine of Gegenqinlian Decoction (GQD) and active components alignment (ACA) in rat plasma using hesperidin as the internal standard (I.S.). Chromatography was performed using a C18 column, with gradient elution with 1% acetic acid-0.001 mol/L ammonium acetate and acetonitrile at 0.2 ml/min. All analytes including I.S. were monitored under positive ionization conditions by selected reaction monitoring with an electrospray ionization source. The optimized mass transition ion-pairs (m/z) for quantitation were 471/297 for puerarin, 471/255 for daidzin, 447/271 for baicalin, 823/453 for glycyrrhizic acid, 419/257 for liquiritin, 336/320 for berberine, 352/336 for palmatine, 338/322 for jateorhizine and 611/303 for hesperidin. The calibration curves were linear over the concentration ranges from 0.15-63.0 to 6.3-6340.0 ng/mL. Intra-day and inter-day precisions (RSD%) were within 15.0%, and accuracy (RE%) ranged from -7.4 to 13.2%. The extraction recoveries were ranged from 60.4 to 93.3%. The proposed method was further applied to compare the pharmacokinetics of all analytes following a single oral administration of GQD and ACA. In conclusion, the eight analytes of GQD and ACA had partly similar pharmacokinetics, which were different from single composition (such as puerarin).
Asunto(s)
Alcaloides/farmacocinética , Cromatografía Liquida/métodos , Medicamentos Herbarios Chinos/farmacocinética , Flavonoides/farmacocinética , Ácido Glicirrínico/farmacocinética , Espectrometría de Masas en Tándem/métodos , Administración Oral , Alcaloides/sangre , Alcaloides/química , Animales , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos/administración & dosificación , Flavonoides/sangre , Flavonoides/química , Ácido Glicirrínico/sangre , Ácido Glicirrínico/química , Límite de Detección , Modelos Lineales , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los ResultadosRESUMEN
An ultra performance liquid chromatography-mass spectrometric (UPLC-MS) method was developed to investigate the pharmacokinetic properties of ephedrine, methylephedrine, amygdalin, and glycyrrhizic acid after oral gavage of Ma Huang Tang (MHT) in Beagles. Beagle plasma samples were pretreated using liquid-liquid extraction, and chromatographic separation was performed on a C18 column using a linear gradient of water-formic acid mixture (0.1%). The pharmacokinetic parameters of ephedrine, methylephedrine, amygdalin, and glycyrrhizic acid from MHT in Beagles were quantitatively determined by UPLC with tandem mass detector. The qualitative detection of the four compounds was accomplished by selected ion monitoring in negative/positive ion modes electrospray ionization-mass spectrometry (ESI-MS). Detection was based on multiple reaction monitoring with the precursor-to-product ion transitions m/z 166.096-116.983 (ephedrine), m/z 179.034-146.087 (methylephedrine), m/z 456.351-323.074 (amygdalin), and m/z 821.606-351.062 (glycyrrhizic acid). The selectivity, sensitivity, linearity, accuracy, precision, extraction recovery, ion suppression, and stability were within the acceptable ranges. The method described was successfully applied to reveal the pharmacokinetic properties of ephedrine, methylephedrine, amygdalin, and glycyrrhizic acid after oral gavage of MHT in Beagles.
Asunto(s)
Amigdalina/sangre , Medicamentos Herbarios Chinos/administración & dosificación , Efedrina/análogos & derivados , Ácido Glicirrínico/sangre , Espectrometría de Masas en Tándem/métodos , Administración Oral , Animales , Cromatografía Líquida de Alta Presión/métodos , Perros , Efedrina/sangre , Límite de DetecciónRESUMEN
Kushen-Gancao Decoction (KGD) is a classic traditional Chinese herb combination in treating viral hepatitis and chronic liver diseases. This study aims to investigate the pharmacokinetic (PK) study of matrine (MT), oxymatrine (OMT), glycyrrhizic acid (GL) and glycyrrhetinic acid (GA) following oral administration of KGD in rats. A rapid, sensitive and reliable HPLC-MS/MS method was successfully developed for the simultaneous determination of MT, OMT, GL and GA in rat plasma. A Inertsil C18 analytical column was used with a gradient mobile phase system of methanol-ammonium acetate (5mM) with a flow rate of 0.5 mL/min. The analysis was performed on a positive and negative ionization electrospray mass spectrometer via multi reaction monitoring (MRM). Linear calibration curves were obtained for the following concentration range: 10-5000 ng/mL for MT, OMT and GL, 50-15,000 ng/mL for GA in rat plasma (R(2)>0.99). The lower limit of quantification (LLOQ) was 5 ng/mL (MT, OMT and GL) and 20 ng/mL (GA). The intra- and inter-day accuracies ranged from -7.91 to 9.10% and precisions (RSD) were within 15%. The analytes were found to be stable under short-term temperature conditions, post-preparative temperature conditions, and after three freeze-thaw cycles conditions. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of KGD.
Asunto(s)
Alcaloides/farmacocinética , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/farmacocinética , Ácido Glicirrínico/farmacocinética , Quinolizinas/farmacocinética , Espectrometría de Masas en Tándem/métodos , Administración Oral , Alcaloides/sangre , Alcaloides/química , Animales , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Ácido Glicirretínico/sangre , Ácido Glicirretínico/química , Ácido Glicirretínico/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/química , Modelos Lineales , Quinolizinas/sangre , Quinolizinas/química , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , MatrinasRESUMEN
The aim of the present study was to evaluate the feasibility of transdermal delivery of glycyrrhizin, an agent used in the treatment of chronic hepatitis C, by cathodal iontophoresis using Ag/AgCl electrodes in vitro. The effects of donor pH (pH 4-7), concentration of drug (0.025-0.2% (w/v)), concentration of external chloride ions (Cl(-)) (0-133 mM), current strength (0-0.5 mA/cm(2)), and permeation enhancers (urea and Tween 80) on the skin permeability of glycyrrhizin were examined in in vitro skin permeation studies using porcine ear skin as the membrane. The cumulative amount of permeated glycyrrhizin and the steady-state skin permeation flux of glycyrrhizin across porcine skin increased in a pH-dependent manner. The skin permeability of glycyrrhizin was independent of the concentration of drug and competed only with a high external Cl(-) concentration. The skin permeation flux of glycyrrhizin increased with the current (R(2)=0.8955). The combination of iontophoresis and enhancers provided an additive or synergistic effect, and a skin permeation flux of about 60 µg/h/cm(2) was achieved. The plasma concentration of glycyrrhizin in humans, extrapolated from the in vitro steady-state permeation flux across porcine skin, was within the therapeutic level. These results suggest that cathodal iontophoresis can be used as a transdermal drug delivery system for glycyrrhizin using reasonable patch sizes and acceptable levels of current intensity.
Asunto(s)
Antiinflamatorios/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Ácido Glicirrínico/administración & dosificación , Iontoforesis/métodos , Piel/metabolismo , Administración Cutánea , Animales , Antiinflamatorios/sangre , Antiinflamatorios/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/farmacocinética , Humanos , Masculino , Polisorbatos/farmacología , Piel/efectos de los fármacos , Absorción Cutánea/efectos de los fármacos , Porcinos , Urea/farmacologíaRESUMEN
The current study aims to investigate the pharmacokinetic properties of Huangqin Tang on different oral doses. An LC-MS method for simultaneous determination of flavonoids and terpenoids in rat plasma was developed and validated. Plasma samples were treated with hydrochloric acid (containing 1% ascorbic acid), precipitated with acetonitrile, separated on a Zorbax SB-C18 column, detected by single quadruple mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The results of specificity, linearity, intra-day and inter-day precisions, accuracy, and stability for LC-MS assay were suitable for the quantification of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid in rat plasma. The concentration-time profiles of baicalin, wogonoside, baicalein, wogonin, oroxylin A and glycyrrhizic acid showed double-peak phenomenon after Huangqin Tang was orally administered at 40 g x kg(-1) dose; all eight constituents in rat plasma showed good dose-exposure relationship within the dosage of 10-40 g x kg(-1); although plasma concentrations were different, the flavonoids with the same backbone showed the similar fate in the body with the corresponding dosage. In conclusion, the LC-MS assay was successfully applied for the pharmacokinetic study of multi-constituents of Huangqin Tang with different doses. Additionally, these constituents demonstrated good pharmacokinetic properties in the body.
Asunto(s)
Medicamentos Herbarios Chinos/química , Flavanonas/sangre , Triterpenos Pentacíclicos/sangre , Administración Oral , Animales , Cromatografía Liquida , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/administración & dosificación , Flavanonas/farmacocinética , Flavonoides/sangre , Flavonoides/farmacocinética , Glucósidos/sangre , Glucósidos/farmacocinética , Ácido Glicirretínico/sangre , Ácido Glicirretínico/farmacocinética , Ácido Glicirrínico/sangre , Ácido Glicirrínico/farmacocinética , Masculino , Monoterpenos/sangre , Monoterpenos/farmacocinética , Triterpenos Pentacíclicos/farmacocinética , Ratas , Ratas Wistar , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
The aim of this study is to investigate the rat intestinal absorption behavior of two main active components, liquiritin, glycyrrhizin and the extract of Glycyrrhiza uralensis. The rat intestinal perfusion model was employed. Concentrations of the compounds of the interest in the intestinal perfusate, bile and plasma samples were determined by HPLC and UPLC. At the same time, the intestinal enzymes incubation test and the partition coefficient determination, the absorption of liquiritin and glycyrrhizin alone and the extract were multiple analyzed. The results showed that the P(eff) (effective permeability) of liquiritin or glycyrrhizin alone or the extract was less than 0.3, which suggested their poor absorption in the intestine. The P(eff) of the two main active components or the extract was not significantly different in duodenum, jejunum, colon and ileum segment. The P(eff) of the glycyrrhizin in the extract had no significant difference in the four intestinal segments compared with the glycyrrhizin alone. The absorption of the liquiritin displayed significant difference (P < 0.05) at ileum segment compared with the liquiritin alone, while it had no markedly change in the other three segments. This phenomenon indicated that some ingredients in the extract might improve the absorption of liquiritin. Moreover, no parent compounds and their metabolites were found in the intestinal perfusate, bile and the plasma samples. The results demonstrated that the influence of the other ingredients in the extract on the two components might not increase the amount of liquiritin and glycyrrhizin in the bile and plasma within the duration of the test.
Asunto(s)
Medicamentos Herbarios Chinos/farmacocinética , Flavanonas/farmacocinética , Glucósidos/farmacocinética , Glycyrrhiza uralensis/química , Ácido Glicirrínico/farmacocinética , Absorción Intestinal , Extractos Vegetales/farmacocinética , Animales , Bilis/metabolismo , Colon/metabolismo , Duodeno/metabolismo , Flavanonas/sangre , Glucósidos/sangre , Ácido Glicirrínico/sangre , Íleon/metabolismo , Yeyuno/metabolismo , Masculino , Raíces de Plantas/química , Plantas Medicinales/química , Ratas , Ratas Sprague-DawleyRESUMEN
The purpose of the current study was to determine whether glycyrrhizin (GL) maintains hepatic glutathione (GSH) levels by inhibiting GSH biliary secretion in normal rats. The effects of glycyrrhizin on hepatic glutathione content, bile flow and biliary secretion of glutathione were examined. Because glutathione is a substrate for multidrug resistance associated protein-2 (Mrp2/ABCC2), the inhibitory effects of GL on Mrp2 in isolated perfused rat liver and in Mrp2-expressing Sf9 membrane vesicles were also examined using the Mrp2 substrate methotrexate (MTX) and estradiol-17-ß-glucuronide (E2 17G). The hepatic content of glutathione in rats following GL perfusion (43.7 µmol/l) in isolated liver perfusion and GL intravenous treatment (25 mg/kg) was significantly higher than that for the control. A marked and dose-dependent decrease in the excretion of glutathione was observed. In addition, the secretion rate of MTX was decreased by 57% in isolated liver perfusion in GL-treated rats. Moreover the ATP-dependent uptake of E2 17G by Mrp2 membrane vesicles was decreased by 75.9% in the 20 µm GL group and by 60.5% in the 2 µm GL group. In conclusion, glycyrrhizin increases hepatic glutathione content possibly through inhibition of Mrp2 which then reduces the biliary excretion of glutathione.
Asunto(s)
Antiinflamatorios/farmacología , Sistema Biliar/metabolismo , Glutatión/metabolismo , Ácido Glicirrínico/farmacología , Hígado/efectos de los fármacos , Animales , Antiinflamatorios/sangre , Bilis/química , Transporte Biológico Activo , Relación Dosis-Respuesta a Droga , Estradiol/análogos & derivados , Estradiol/farmacocinética , Ácido Glicirrínico/sangre , Técnicas In Vitro , Hígado/metabolismo , Masculino , Metotrexato/farmacocinética , Metotrexato/farmacología , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/antagonistas & inhibidores , Perfusión , Ratas , Ratas Sprague-Dawley , Especificidad por SustratoRESUMEN
Si-Ni-San (SNS) is a widely used traditional Chinese medicine formula (TCMF) in treating various diseases. However, the in vivo integrated metabolism of its multiple components remains unknown. In this paper, a liquid chromatography coupled with diode array detection and triple-quadrupole spectrometry (LC-DAD-MS/MS) method was developed for detection and identification of SNS metabolites in rat plasma and urine at a normal clinical dosage. Accurate structural elucidation was performed using MS/MS, UV data and n-octanol/water partition coefficient. Based on the proposed strategy, 36 absorbed compounds and 29 metabolites in plasma and 33 metabolites in urine were detected by a highly sensitive MRM method. Our results indicated that phase II reactions (e.g., methylation, glucuronidation and sulfation) were the main metabolic pathways of gallic acid and flavanones, while phase I reactions (e.g., hydroxylation) were the major metabolic reaction for triterpenoid saponins. The metabolite profile analysis of SNS provided a comprehensive understanding of the in vivo metabolic fates of constituents in SNS. Moreover, the results in this work demonstrated the present strategy based on the combination of chromatographic, spectrophotometric, mass-spectrometric, and software prediction to detect and identify metabolites was effective and reliable. And such a strategy may also be extended to investigate the metabolism of other TCMF.