likviriton and liquiritigenin

The inhibitory effects of liquiritigenin, liquiritin and glycyrrhizic acid against the hazards during the preparation of thermal reaction beef flavoring were investigated using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Liquiritigenin(1.5 mM) inhibited N

Topics: Animals; Cattle; Free Radicals; Glycation End Products, Advanced; Glycyrrhizic Acid; Lysine; Meat; Pyruvaldehyde; Tandem Mass Spectrometry

Licorice is an ancient food and medicinal plant. Liquiritigenin and liquiritin, two kinds of major flavonoes in licorice, are effective substances used as antioxidant, anti-inflammatory and tumor-suppressive food, cosmetics or medicines. However, their in vivo metabolites have not been fully explored.. To clarify the metabolism of liquiritigenin and liquiritin in mice.. In this study, we developed a liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry approach to determine the metabolites in mice plasma, bile, urine and feces after oral administration of liquiritigenin or liquiritin. The structures of those metabolites were tentatively identified according to their fragment pathways, accurate masses, characteristic product ions, metabolism laws or reference standard matching.. A total of 26 and 24 metabolites of liquiritigenin or liquiritin were respectively identified. The products related with apigenin, luteolin or quercetin were the major metabolites of liquiritigenin or liquiritin in mice. Seven main metabolic pathways including (de)hydrogenation, (de)hydroxylation, (de)glycosylation, (de)methoxylation, acetylation, glucuronidation and sulfation were summarized to tentatively explain their biotransformation.. This study not only can provide the evidence for in vivo metabolites and pharmacokinetic mechanism of liquiritigenin and liquiritin, but also may lay the foundation for further development and utilization of liquiritigenin, liquiritin and then licorice.

Topics: Administration, Oral; Animals; Bile; Biotransformation; Chromatography, High Pressure Liquid; Drug Elimination Routes; Feces; Flavanones; Glucosides; Glycyrrhiza; Male; Metabolomics; Mice, Inbred C57BL; Plant Extracts; Tandem Mass Spectrometry

Previously, we have investigated the therapeutic mechanism of Qingzao Jiufei Decoction (QZJFD), a Chinese classic prescription, on acute lung injury (ALI), however, which remained to be further clarified together with the underlying efficacy related compounds for quality markers (Q-markers).. To explore Q-markers of QZJFD on ALI by integrating a stepwise multi-system with 'network pharmacology-metabolomics- pharmacokinetic (PK)/ pharmacodynamic (PD) modeling'.. First, based on in vitro and in vivo component analysis, a network pharmacology strategy was developed to identify active components and potential action mechanism of QZJFD on ALI. Next, studies of poly-pharmacology and non-targeted metabolomics were used to elaborate efficacy and verify network pharmacology results. Then, a comparative PK study on active components in network pharmacology was developed to profile their dynamic laws in vivo under ALI, suggesting Q-marker candidates. Next, quantified analytes with marked PK variations after modeling were fitted with characteristic endogenous metabolites along drug concentration-efficacy-time curve in a PK-PD modeling to verify and select primary effective compounds. Finally, Q-markers were further chosen based on representativeness among analytes through validity analysis of PK quantitation of primary effective compounds.. In virtue of 121 and 33 compounds identified in vitro and in vivo, respectively, 33 absorbed prototype compounds were selected to construct a ternary network of '20 components-47 targets-113 pathways' related to anti-ALI of QZJFD. Predicted mechanism (leukocytes infiltration, cytokines, endogenous metabolism) were successively verified by poly-pharmacology and metabolomics. Next, 18 measurable components were retained from 20 analytes by PK comparison under ALI. Then, 15 primary effective compounds from 18 PK markers were further selected by PK-PD analysis. Finally, 9 representative Q-markers from 15 primary effective compounds attributed to principal (chlorogenic acid), ministerial (methylophiopogonanone A, methylophiopogonanone B), adjuvant (sesamin, ursolic acid, amygdalin), conductant drugs (liquiritin apioside, liquiritigenin and isoliquiritin) in QZJFD, were recognized by substitutability and relevance of plasmatic concentration at various time points.. 9 Q-markers for QZJFD on ALI were identified by a stepwise integration strategy, moreover, which was a powerful tool for screening Q-makers involved with the therapeutic action of traditional Chinese medicine (TCM) prescription and promoting the process of TCM modernization and scientification.

Topics: Acute Lung Injury; Administration, Oral; Amygdalin; Animals; Biological Availability; Biomarkers, Pharmacological; Chalcone; Chlorogenic Acid; Dioxoles; Drugs, Chinese Herbal; Flavanones; Glucosides; Lignans; Male; Metabolomics; Rats, Wistar; Triterpenes; Ursolic Acid

Hepatic sinusoidal obstruction syndrome (HSOS) is a life-threatening liver disease caused by the damage to liver sinusoidal endothelial cells (LSECs). Liquiritigenin and liquiritin are two main compounds in Glycyrrhizae Radix et Rhizoma (Gan-cao). Our previous study has shown that both liquiritigenin and liquiritin alleviated monocrotaline (MCT)-induced HSOS in rats via inducing the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant signaling pathway. This study aims to further investigate whether inhibiting liver inflammatory injury also contributed to the liquiritigenin and liquiritin-provided alleviation on MCT-induced HSOS. The results of serum alanine/aspartate aminotransferases (ALT/AST) activities and total bilirubin (TBil) amount, liver histological evaluation, scanning electron microscope observation and hepatic metalloproteinase-9 (MMP9) expression showed that liquiritigenin and liquiritin both alleviated MCT-induced HSOS in rats. Liquiritigenin and liquiritin reduced the increased liver myeloperoxidase (MPO) activity, mRNA expression of pro-inflammatory factors, hepatic infiltration of immune cells, hepatic toll-like receptor 4 (TLR4) expression and nuclear factor κB (NFκB) nuclear accumulation induced by MCT in rats. Furthermore, liquiritigenin and liquiritin attenuated MCT-induced liver mitochondrial injury, increased the decreased Lon protein expression and reduced the release of heat shock protein 60 (HSP60). Moreover, liquiritigenin and liquiritin also reduced NFκB nuclear accumulation and decreased the elevated cellular mRNA expression of NFκB-downstream pro-inflammatory cytokines induced by HSP60 in macrophage RAW264.7 cells. In conclusion, our study revealed that both liquiritigenin and liquiritin alleviated MCT-induced HSOS by inhibiting hepatic inflammatory responses triggered by HSP60.

Topics: Adenosine Triphosphate; Animals; Anti-Inflammatory Agents; Chaperonin 60; Flavanones; Glucosides; Hepatic Veno-Occlusive Disease; Liver; Male; Mice; Mitochondria; Monocrotaline; Rats, Sprague-Dawley; RAW 264.7 Cells; Reactive Oxygen Species

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.

Topics: Administration, Oral; Animals; Chalcones; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Male; Plant Extracts; Quality Control; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Cyclophosphamide (CPA) is a chemotherapeutic drug widely used in the treatment of breast cancer or leukemia in clinic. However, CPA was reported to have hepatotoxicity. This study aims to observe the engaged mechanism of CPA-induced liver injury in mice and the protection of liquiritin (LQ) and liquiritigenin (LG). Liver sinusoidal endothelial injury induced by CPA (20, 40 mg/kg) in mice was evidenced by the elevated hepatic metalloproteinase-9 (MMP-9) expression, and the results from liver histological evaluation and scanning electron microscope observation. CPA increased hepatic infiltration of neutrophils, liver myeloperoxidase (MPO) activity, serum interleukin-6 (IL-6) content, hepatic IL-6 mRNA expression, toll-like receptor-4 (TLR4) expression and nuclear factor κB (NFκB) activation in mice. Elevated serum contents of damage associated molecular patterns (DAMPs) including high mobility group box 1 (HMGB1), heat shock protein 60 (HSP60) and glucose-regulated protein 94 (Grp94) were found in mice treated with CPA. Liver sinusoidal endothelial injury and inflammation induced by CPA were diminished in TLR4 knock-out mice. LG and LQ (40, 80 mg/kg) both ameliorated liver sinusoidal endothelial injury, and reduced the increased hepatic infiltration of neutrophils, MPO activity, hepatic IL-6 mRNA expression and NFκB activation induced by CPA. In summary, these results indicate that TLR4-NFκB-mediated inflammatory injury initiated by DAMPs was critically involved in CPA-induced hepatotoxicity. LG and LQ alleviated CPA-induced liver sinusoidal endothelial injury and inflammatory injury in mice.

Topics: Alarmins; Animals; Chemical and Drug Induced Liver Injury; Cyclophosphamide; Endothelial Cells; Flavanones; Glucosides; Inflammation; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Signal Transduction; Toll-Like Receptor 4

Topics: Administration, Oral; Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Female; Flavanones; Glucosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Monoterpenes; Polycystic Ovary Syndrome; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Semen Strychni is known for its treatment of rheumatic arthritis with a low therapeutic index. Liquorice contributes a lot in herb detoxification according to the traditional Chinese medicine theory. A simple, rapid, and sensitive liquid chromatography-mass spectrometric method (LC-MS) was developed and validated for simultaneous determination of main bioactive ingredients in liquorice and Semen Strychni in rat plasma. Using moclobemide and cyproterone acetate as the internal standards, the analytes were pretreated via protein precipitation with methanol. An Ultimate AQ-C18 column (3.0 μm, 3.0 × 100 mm) was employed for chromatographic separation, combining with gradient elution. The mobile phase consisted of 0.07% formic acid and 0.12% ammonium acetate in aqueous phase (A) and acetonitrile in organic phase (B). The elution program was as follows: 0-0.5 min, 20% B; 0.5-1 min, 20-60% B; 1-7 min, 60-85% B; and 7-7.5 min, returned to 20% B, then continued to 12 min. Selected reaction monitoring was performed in both positive and negative ESI. Positive mode was adopted for detection of strychnine, brucine, and moclobemide, while negative mode was used for glycyrrhizic acid, glycyrrhetinic acid, liquiritigenin, isoliquiritigenin, liquiritin, and cyproterone acetate. The method was validated for specificity, linearity, matrix effect, recovery, precision, accuracy, and stability. The results show that this method is sensitive, accurate and robust for biological matrix analysis. Moreover, the proposed method was applied to a pharmacokinetic study in Sprague-Dawley rats for investigating the mechanism of which liquorice detoxifies Semen Strychni.

Topics: Animals; Chromatography, Liquid; Flavanones; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Plasma; Rats; Reproducibility of Results; Semen; Strychnine

Hepatic sinusoidal obstruction syndrome (HSOS) is a serious and life-threatening liver disease. Liquiritigenin (LG) and liquiritin (LQ) are natural flavonoids distributed in Glycyrrhizae Radix et Rhizoma (Gan-cao). This study aims to investigate the protective effect and mechanism of LG and LQ against monocrotaline (MCT)-induced HSOS. Results of serum alanine/aspartate aminotransferases (ALT/AST) activities, liver histological evaluation and scanning electron microscope observation, and hepatic metalloproteinase-9 (MMP-9) expression demonstrated that LG and LQ both alleviated HSOS induced by MCT in rats. Results of hepatic reactive oxygen species (ROS), malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), oxidized glutathione (GSSG) and reduced glutathione (GSH) contents, glutathione reductase (GR) and superoxide dismutase (SOD) activities showed that LG and LQ attenuated MCT-induced liver oxidative stress injury. Furthermore, LG and LQ were found to promote Nrf2 nuclear translocation and lead to the increased expression of Nrf2 downstream antioxidative genes. Molecule docking analysis indicated the potential interaction of LG and LQ with Nrf2 binding site in the kelch-like ECH-associated protein-1 (Keap1) protein. Finally, Nrf2 knock-out mice were used. The results showed that LG and LQ both alleviated MCT-induced HSOS in wild-type mice, but such protection was totally diminished in Nrf2 knock-out mice. In conclusion, our study revealed that LG and LQ alleviated MCT-induced HSOS by inducing the activation of hepatic Nrf2 antioxidative defense system.

Topics: Animals; Antioxidants; Flavanones; Glucosides; Hepatic Veno-Occlusive Disease; Hypolipidemic Agents; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Docking Simulation; Molecular Structure; Monocrotaline; NF-E2-Related Factor 2; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species

Jiegeng Gancao decoction, which is composed of Jiegeng and Gancao at a weight ratio of 1:2, was widely used for treating pharyngalgia and cough for thousands of years. Our previous work indicated that Gancao could increase the systemic exposure of platycodin D and deapio-platycodin D, two main components in Jiegeng. However, whether Jiegeng could alter the pharmacokinetics of the main compounds in Gancao is still unknown. Thus, the purpose of this study was to compare the oral pharmacokinetics of flavonoids and saponins from Gancao alone vs. after co-administration with Jiegeng. Furthermore, Caco-2 cell transport and fecal hydrolysis were investigated to explain the altered pharmacokinetic properties. Pharmacokinetics results suggested that the bioavailability of liquiritin, isoliquiritin, glycyrrhizin and its metabolite, glycyrrhetinic acid, could be improved while bioavailability of liquiritigenin and isoliquiritigenin deteriorated when co-administered with Jiegeng. The Caco-2 transport study showed no significant difference of the P

Topics: Caco-2 Cells; Chalcone; Flavanones; Flavonoids; Glucosides; Glycyrrhiza uralensis; Humans; Saponins; Triterpenes

Licorice is one of the most common herbs in traditional Chinese medicine, and classified as top\ grade in Shen Nong Ben Cao Jing. There are three different original plants of licorice stipulated in Chinese\ Pharmacopeia, Glycyrrhiza uralensis Fisch., Glycyrrhiza glabra L., and Glycyrrhiza inflata Bat. However,\ previous investigation showed that the pharmacodynamic effects of the three licorices were quite different. It is\ very difficult to identify them by the classical identification methods. In order to establish a fast and effective\ identification method, we collected 240 licorice plants from 21 populations of 7 provinces, and amplified their\ ITS and psbA-trnH sequences. ITS sequences with a full length of 616 bp and psbA-trnH sequences with a full\ length of 389 bp were obtained separately. Using DNAMAN to analyze these sequences, 4 variable sites were\ found in ITS sequences and 2 ITS haplotypes were determined, and 3 variable sites were found in psbA-trnH\ sequences and 4 psbA-trnH haplotypes were determined. With the combination analysis of ITS and psbA-trnH\ sequences, the molecular identification method of original licorice was established. Using this method, 40\ samples of licorice slices collected from 4 main herbal material markets in China were identified successfully.\ Furthermore, the contents of 2 triterpenes, 18α-glycyrrhizic acid and 18β-glycyrrhizic acid, and 4 flavonoids,\ liquiritin, isoliquiritin, liquiritigenin, and isoliquiritigenin in these licorice pieces were examined by HPLC and\ the results were analyzed using SPSS 21.0. This study provides a new method in identification of licorice,\ which may serve as a guideline for quality control of licorice slices.

Topics: Chalcone; Chalcones; China; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhiza; Glycyrrhiza uralensis; Glycyrrhizic Acid; Triterpenes

Licorice (Glycyrrhizae radix), the root of Glycyrrhiza uralensis Fisch. (Leguminosae), is mainly used to moderate the characteristics of toxic herbs in Traditional Chinese Medicine, which could be partly interpreted as detoxification. However, the underlying mechanism is still not fully elucidated. Nuclear factor erythroid 2-related factor 2 (Nrf2) plays a key role in the protection against toxic xenobiotics. In our previous research, we have identified that extracts from Glycyrrhiza uralensis induced the expression of Nrf2 nuclear protein and its downstream genes. This research aims to screen the most potent Nrf2 inducer isolated from Glycyrrhiza uralensis and examine its effect on Nrf2 signaling pathway and detoxification system.. Four compounds derived from Glycyrrhiza uralensis (glycyrrhetinic acid, liquiritigenin, isoliquiritigenin and liquiritin) were screened by ARE-luciferase reporter. The most potent ARE-luciferase inducer was chosen to further examine its effect on Nrf2 and detoxification genes in HepG2 cells. The role of Nrf2-dependent mechanism was tested by using Nrf2 knockout mice (Nrf2 KO) and Nrf2 wild-type mice (Nrf2 WT).. ARE-luciferase reporter assay showed these four compounds were all potent Nrf2 inducers, and isoliquiritigenin was the most potent inducer. Isoliquiritigenin significantly up-regulated the expression of Nrf2 and its downstream detoxification genes UDP-glucuronosyltransferase 1A1 (UGT1A1), glutamate cysteine ligase (GCL), multidrug resistance protein 2 (MRP2) and bile salt export pump (BSEP) in vitro and in vivo. Additionally, isoliquiritigenin showed Nrf2-dependent transactivation of UGT1A1, GCLC and MRP2.. Isoliquiritigenin, isolated from Glycyrrhiza uralensis, stimulates detoxification system via Nrf2 activation, which could be a potential protective mechanism of licorice.

Topics: Animals; Antioxidant Response Elements; ATP Binding Cassette Transporter, Subfamily B, Member 11; ATP-Binding Cassette Transporters; Chalcones; Female; Flavanones; Glucosides; Glucuronosyltransferase; Glutamate-Cysteine Ligase; Glycyrrhetinic Acid; Glycyrrhiza uralensis; Hep G2 Cells; Humans; Luciferases, Renilla; Mice, Inbred ICR; Mice, Knockout; Multidrug Resistance-Associated Protein 2; Multidrug Resistance-Associated Proteins; NF-E2-Related Factor 2; RNA, Messenger

Fighting against multidrug-resistant bacteria requires reliable methods to evaluate the effect of antibacterial agents. As a universal, non-destructive, and highly sensitive tool, microcalorimetry has been used in many biological investigations to provide continuous real-time monitoring of the metabolic activity. This method, based on heat-flow output, was used to evaluate the influence of two flavonoid compounds (liquiritigenin and liquiritin) on Escherichia coli. Some crucial information, such as the thermogenic power-time curve and thermokinetic parameters of E. coli growth affected by the two compounds, was obtained and further studied by chemometric techniques including similarity analysis, multivariate analysis of variance, and principal component analysis. By comparing the values of two main parameters, k 2 (growth rate constant of the second exponential phase) and Q 1 (heat output of the first exponential growth phase) of E. coli based on the box and whisker plot, liquiritigenin and liquiritin could be differentiated according to their antibacterial effects; liquiritin with IC50 (half-inhibitory concentration) of 198.6 μg mL(-1) expressed a stronger antibacterial effect than liquiritigenin with IC50 of 337.8 μg mL(-1). The glucoside group in liquiritin containing four additional free hydroxyls in the diphenylpropane skeleton was crucial for inducing the antibacterial effect. Liquiritin might be a promising candidate against E. coli. This study provides a valuable method for searching for novel antibacterial agents using microcalorimetry with chemometrics.

Topics: Anti-Bacterial Agents; Calorimetry; Escherichia coli; Flavanones; Glucosides; Inhibitory Concentration 50; Microbial Sensitivity Tests

This study provides the scientific basis for the anti-inflammatory effects of licorice extract in a t-BHP (tert-butyl hydrogen peroxide)-induced liver damage model and the effects of its ingredients, glycyrrhizic acid (GA), liquiritin (LQ) and liquiritigenin (LG), in a lipopolysaccharide (LPS)-stimulated microglial cell model. The GA, LQ and LG inhibited the LPS-stimulated elevation of pro-inflammatory mediators, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and interleukin (IL)-6 in BV2 (mouse brain microglia) cells. Furthermore, licorice extract inhibited the expression levels of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) in the livers of t-BHP-treated mice models. This result suggested that mechanistic-based evidence substantiating the traditional claims of licorice extract and its three bioactive components can be applied for the treatment of inflammation-related disorders, such as oxidative liver damage and inflammation diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Cell Line; Disease Models, Animal; Flavanones; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Inflammation; Inflammation Mediators; Liver; Male; Mice; Mice, Inbred ICR; Nitric Oxide; Oxidative Stress; Plant Extracts; Tumor Necrosis Factor-alpha

A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous determination of seven constituents including puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma using schisandrin as the internal standard (IS). The plasma samples were pretreated by a one-step direct protein precipitation with acetonitrile. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and water (containing 0.1% formic acid and 5mM ammonium acetate). All analytes and IS were quantitated through electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. The mass transitions were as follows: m/z 417.5→297.2 for puerarin, m/z 417.1→255.2 for daidzin, m/z 255.2→152.4 for daidzein, m/z 498.1→179.3 for paeoniflorin, m/z 481.1→197.3 for albiflorin, m/z 436.2→257.3 for liquiritin, m/z 257.2→137.3 for liquiritigenin and m/z 415.0→384.2 for IS, respectively. All calibration curves exhibited good linearity (r>0.9979) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at three different levels were both less than 14.3% and the accuracies (RE) ranged from -13.2% to 14.8%. The extraction recoveries of the seven compounds ranged from 72.9% to 117.4%. The validated method was successfully applied to pharmacokinetic study of the seven components in female rat plasma after oral administration of Ge-Gen Decoction aqueous extract.

Topics: Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Isoflavones; Monoterpenes; Rats; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Wild or cultivated Glycyrrhiza uralensis FISCHER (G. uralensis) are the main source of licorice, and they contain the similar compounds, such as the triterpenoid saponins and flavonoids, but above two kinds of the components contents are low level in the cultivated licorice. To produce the high quality cultivated licorices, researchers studied the affecting factors about the compounds producing in the plant of licorice, and then found that the growth years, genetic differences and water deficit are all the important factors. In this paper, we found that there were different distribution patterns of the main five active components (FAC) including glycyrrhizin, liquiritin, isoliquiritin, liquiritigenin and isoliquiritigenin in the taproot and stolon of G. uralensis and maybe they are also important influence factors to the FAC contents of the licorices. In wild G. uralensis, the contents of FAC tended to be lower in the younger parts of the stolon, and in the cultivated G. uralensis taproot, the contents of glycyrrhizin, liquiritin and isoliquiritin tended to increase from top to end, contrary to the contents of liquiritigenin and isoliquiritigenin, which increased first and then decreased. Our results will contribute to the analyses of factors which influence the quality of licorice, and provide some reference for cultivating high quality licorices for herbal medicine.

Topics: Chromatography, High Pressure Liquid; Flavanones; Glucosides; Glycyrrhiza uralensis; Glycyrrhizic Acid; Isomerism; Limit of Detection; Molecular Structure; Plant Roots; Plant Shoots; Species Specificity

To study the permeation of liquiritigenin (LQG) and liquiritin (LQ) as licorice flavonoids into the skin, we prepared ceramide liposome-in-cellulose hydrogel complex system.. Liposome-in-hydrogel complex systems were developed by incorporating ceramide liposomes into cellulose hydrogels by the swelling method. We evaluated their physical and chemical properties, encapsulation efficiency and skin permeability using Franz Diffusion Cell. It was visually seen by CLSM images analysis.. The ceramide liposome, consisting of biocompatible lipid membranes, remained stable for over 3 weeks. Encapsulation efficiencies for liquiritigenin and liquiritin-loaded liposome-in-hydrogel were 69.39% and 64.71%, respectively. Liposome-in-hydrogel complex systems (LQG: 56.55%, LQ: 66.99%) had greater skin permeability than control (LQG: 4.92%, LQ: 5.30%) or a single liposome systems (LQG: 43.34%, LQ: 48.97%) and hydrogel systems (LQG: 38.21%, LQ: 55.07%).. Liposome-in-hydrogel system can be a potential drug delivery system for topical delivery of antioxidants such as licorice flavonoids to construct antioxidative skin barrier.

Topics: Administration, Cutaneous; Drug Delivery Systems; Epidermis; Flavanones; Glucosides; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate; Liposomes; Microscopy, Confocal; Permeability

A reversed phase chromatographic system, composed of a stationary phase of C18 silica gel (ODS, 20μm) and a mobile phase of ethanol/water, was used to separate liquiritin and liquiritigenin in the raw material of flavonoids. The linear adsorption isotherm and the equilibrium-dispersive model were adopted to approximatively describe the chromatographic separation behaviors of liquiritin and liquiritigenin in the raw material under different column temperatures, ethanol contents and flow rates of the mobile phase, sample concentrations and feeding times. Combined with orthogonal design, the ED model was used to optimize the chromatographic separating conditions, the corresponding experimental result with a good agreement was obtained and the overload separation was realized.

Topics: Chromatography, High Pressure Liquid; Computer Simulation; Ethanol; Flavanones; Glucosides; Glycyrrhiza; Temperature; Water

The detailed mechanisms on licorice-drug interaction remain to be unclear. The aim of the present study is to investigate the inhibition of important UGT isoforms by two important ingredients of licorice, liquiritin, and liquiritigenin. The results showed that liquiritigenin exhibited stronger inhibition towards all the tested UGT isoforms than liquiritin. Data fitting using Dixon and Lineweaver-Burk plots demonstrated the competitive inhibition of liquiritigenin towards UGT1A1 and UGT1A9-mediated 4-MU glucuronidation reaction. The inhibition kinetic parameters (Ki ) were calculated to be 9.1 and 3.2 μM for UGT1A1 and UGT1A9, respectively. Substrate-dependent inhibition behaviour was also observed for UGT1A1 in the present study. All these results will be helpful for understanding the deep mechanism of licorice-drug interaction. However, when translating these in vitro parameters into in vivo situations, more complex factors should be considered, such as substrate-dependent inhibition of UGT isoforms, the contribution of UGT1A1 and UGT1A9 towards the metabolism of drugs, and many factors affecting the abundance of ingredients in the licorice.

Topics: Flavanones; Food-Drug Interactions; Glucosides; Glucuronosyltransferase; Glycyrrhiza; Humans; Hymecromone; Isoenzymes; Kinetics; UDP-Glucuronosyltransferase 1A9

Shakuyakukanzoto (SKT) composed of Glycyrrhizae radix (G. radix) and Paeoniae radix (P. radix) has been traditionally used in Japan, Korea and China as an antispasmodic drug for the treatment of skeletal muscle cramps and intestinal cramps.. To evaluate the antispasmodic activity of SKT and its two components, as well as to identify the key constituents of the components which mediate this effect in skeletal muscles in vivo.. An experimental cramp model was constructed to evaluate the effects of peripherally-acting muscle relaxants on electrically-induced cramps under physiological conditions. This was accomplished by surgically isolating the motor supply to the gastrocnemius muscle in an anesthetized rat and delivering electrical stimuli to an isolated tibial nerve to induce tetanic contractions. We first tested dantrolene, a well-known peripherally-acting relaxant, to determine the sensitivity and reliability of our experimental model. We then evaluated the effects of SKT, P. radix, G. radix, and the eight active constituents of G. radix against tetanic contractions.. We found that dantrolene (10 and 30 mg/kg, i.d.) rapidly and significantly inhibited tetanic contractions (P<0.01) irrespective of dose. SKT (0.5, 1.0, and 2.0 g/kg, i.d.) and G. radix (0.5 and 1.0 g/kg, i.d.) also significantly inhibited tetanic contractions (P<0.01) but in a dose-dependent manner owing to the actions of six of the eight active constituents in G. radix (liquiritin apioside, liquiritigenin, isoliquiritin apioside, isoliquiritigenin, glycycoumarin, and glycyrrhetinic acid, 20 μmol/kg, i.v.). These constituents, which include flavonoids, a triterpenoid, and a courmarin derivative, demonstrated temporal variations in their inhibitory activity. In contrast, P. radix (0.5 and 1.0 g/kg, i.d.) did not show a statistically significant antispasmodic effect in our study; however, we previously found that it had a significant antinociceptive effect.. Our findings show that SKT inhibits tetanic contractions in vivo and that G. radix is the main antispasmodic component due to the actions of its active constituents, thus supporting the traditional use of SKT. We further propose that SKT containing the antispasmodic G. radix and antinociceptive P. radix is a pharmaceutically elegant option for muscle cramps as treatment requires a two-pronged approach, i.e., inhibition of hyperexcitable skeletal tissues and modulation of the pain accompanying cramps.

Topics: Animals; Animals, Outbred Strains; Chalcone; Chalcones; Coumarins; Dantrolene; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Combinations; Drugs, Chinese Herbal; Electric Stimulation; Flavanones; Glucosides; Glycyrrhetinic Acid; Glycyrrhiza; Male; Mice; Muscle Contraction; Muscle Cramp; Muscle, Skeletal; Paeonia; Parasympatholytics; Phytotherapy; Plant Roots; Rats; Rats, Wistar; Rotarod Performance Test; Tibial Nerve

A new, simple, accurate and reliable full-time five-wavelength fusion method for the simultaneous separation and determination of nine active chemical compositions (liquiritin apioside, liquiritin, isoliquiritin apioside, ononin, isoliquiritin, liquiritigenin, calycosin, isoliquiritigenin, Glycyrrhizic acid monoammonium salt) in traditional Chinese medicine Glycyrrhiza was developed using reverse phase high-performance liquid chromatography (RP-HPLC) coupled with a diode-array detector (DAD). The chromatographic separation was performed on an Agilent TC-C18 column with gradient elution using 0.04% methanoic acid (A) and acetonitrile (B) at a flow rate of 1.0 mL min(-1) and UV detection at 248 nm, 250 nm, 276 nm, 362 nm, 370 nm. The standard curves were linear over the range of 2.1379-12.8272 μg for liquiritin apioside, 3.9299-23.5794 μg for liquiritin, 1.0432-6.2592 μg for isoliquiritin apioside, 0.8764-5.8584 μg for ononin, 1.0701-6.4205 μg for isoliquiritin, 1.3685-8.2111 μg for liquiritigenin, 0.3927-2.3563 μg for calycosin, 0.2498- 1.4986 μg for isoliquiritigenin, 2.0094-12.0564 μg for Glycyrrhizic acid monoammonium salt, respectively (r(2) > 0.9997). The recoveries and relative standard deviation (RSD) varied from 95.09% to 103.54% and 1.09% to 2.36%, respectively. The precision for all the analytes was less than 2.52%. The method indicated good performance in terms of precision, accuracy and linearity. The method enabled the simultaneous determination of nine active chemical compositions for quality control of Glycyrrhiza.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Isoflavones; Quality Control

Sinisan is a widely used traditional Chinese medicine (TCM) in treating various diseases; however, the in vivo metabolic profile of its multiple components remains unknown. In this paper, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to identify the metabolites of Sinisan extract in rat plasma, urine, feces and bile after intragastric administration. Using MS(E) and mass defect filter techniques, 41 metabolites of 10 parent compounds (naringin, naringenin, hesperidin, neohesperidin, liquiritin, liquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, saikosaponin a and saikosaponin d) were detected and tentatively identified. It was shown by our results that these compounds was metabolized to the forms of hydroxylation, glucuronidation, sulfation, glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Hesperidin; Hydroxylation; Male; Plants, Medicinal; Random Allocation; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Licorice (the root of Glycyrrhizae plant) has been used as an oriental herbal medicine for thousands of years. The licorice flavonoid components are reported to possess immunomodulatory activities. In this present study, we investigated the immunomodulatory effects of liquiritigenin (LG) and liquiritin (LQ), licorice flavonoid components, against disseminated candidiasis due to Candida albicans, a dimorphic fungus, that causes severe disease via hematogenous dissemination and local diseases such as vaginitis and thrush. Results showed that direct interaction of LG or LQ with C. albicans yeast cells resulted in no growth-inhibition, in vitro. When tested in a murine model of disseminated candidiasis, mice given LQ intraperitoneally before intravenous challenge with live C. albicans yeast cells had similar mean survival times (MST) as untreated mice groups. On the contrary, mice given LG in the same manner as LQ above had longer MST than the untreated mice groups (P < 0.05). In one experiment, 3 out of 5 LG-treated mice survived during the entire period of the 55-day observation. Furthermore, the 3 survivors were cured -- shown by a lack of CFU (colony forming unit) in the kidneys. This protection was nulled when mice were pretreated with anti-CD4+ antibody before LG-treatment and challenge with the yeast. However, the protection was transferable by the CD4+ T cells isolated from LG-treated mice not infected with the yeast. In addition, mice given CD4+ T cells that were pre-treated with LG, in vitro were also protected against disseminated candidiasis. ELISA analysis revealed that in LG-treated mice IFNgamma and IL-2 were dominantly produced compared to IL-4 and IL-10. When LG-given mice were treated with anti-mouse IFNgamma, the protection was again nulled. Combined together, these results indicate that LG protects mice against disseminated candidiasis by the CD4+ Th1 immune response.

Topics: Adoptive Transfer; Animals; Anti-Infective Agents; Antibodies, Blocking; Candida albicans; Candidiasis; Colony Count, Microbial; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Flavanones; Glucosides; Glycyrrhiza; Immunity, Cellular; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Phytotherapy; Plant Roots; Th1 Cells

Glycyrrhizae radix has been used as one of the oldest and most frequently employed botanicals in both western and oriental countries. Previously, we showed that liquiritigenin (LQ), an aglycone of liquiritin in G. radix, exerts cytoprotective effects against heavy metal-induced toxicity in vitro. This study investigated in vivo protective effects of LQ against acute liver injuries induced by acetaminophen (APAP) or APAP plus buthionine sulfoximine (BSO). Liver injuries were assessed by blood biochemistry and histopathology in rats administered with LQ purified from the acid hydrolyates of liquiritin singly (p.o. or i.v., 2-4 days) or in combination with dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylenedioxybiphenyl-2,2'-dicarboxylate (DDB), a synthetic derivative of Schisandrin C in Fructus shizandrae, and exposed to APAP or APAP + BSO. LQ treatments (oral) effectively decreased liver injuries induced by a single dose of APAP, as evidenced by decreases in hepatic necrosis and inflammation as well as plasma alanine aminotransferase and lactate dehydrogenase activities. LQ, when intravenously applied, enhanced hepatoprotective effect with a greater potency. APAP + BSO led to severe liver injuries, resulting in lethality. LQ pretreatments significantly reduced the potentiated liver necrosis, decreasing mortality. In spite of the improvement in blood biochemistry, DDB failed to protect the liver from injuries induced by APAP or APAP + BSO. Combined treatments of rats with LQ and DDB showed some additive protective effect. The present study demonstrates that LQ efficaciously protects the liver from acute injuries induced by APAP or from APAP-induced severe injuries during GSH deficiency, indicating that LQ is one of the principal cytoprotective components comprised in G. radix.

Topics: Acetaminophen; Acute Disease; Alanine Transaminase; Animals; Buthionine Sulfoximine; Chromatography, High Pressure Liquid; Fabaceae; Flavanones; Glucosides; L-Lactate Dehydrogenase; Liver; Molecular Structure; Rats; Rats, Sprague-Dawley