wogonin and 5-7-dihydroxy-6-methoxy-2-phenylchromen-4-one

Huanglian Jiedu Decoction (HLJDD) is a Traditional Chinese Medicine (TCM) formula comprising four herbal medicines. This decoction has long been used in China for clinically treating T2DM. However, the molecular mechanism of HLJDD treat for T2DM is still not fully known. Hence, this study was designed to reveal the synergistic mechanism of HLJDD formula in the treatment of T2DM by using network pharmacology method and molecular docking.. Retrieving and screening of active components of different herbs in HLJDD and corresponding T2DM-related target genes across multiple databases. Subsequently, STRING and Cytoscape were applied to analysis and construct PPI network. In addition, cluster and topological analysis were employed for the analysis of PPI networks. Then, the GO and KEGG enrichment analysis were performed by using ClueGO tool. Finally, the differentially expressed analysis was used to verify whether the expression of key target genes in T2DM and non-T2DM samples was statistically significant, and the binding capacity between active components and key targets was validated by molecular docking using AutoDock.. There are 65 active components involved in 197 T2DM-related targets that are identified in HLJDD formula. What is more, 39 key targets (AKT1, IL-6, FOS, VEGFA, CASP3, etc.) and 3 clusters were obtained after topological and cluster analysis. Further, GO and KEGG analysis showed that HLJDD may play an important role in treating T2DM and its complications by synergistically regulating many biological processes and pathways which participated in signaling transduction, inflammatory response, apoptotic process, and vascular processes. Differentially expressed analysis showed that AKT1, IL-6, and FOS were upregulated in T2DM samples and a significant between sample differential expression. These results were validated by molecular docking, which identified 5 high-affinity active components in HLJDD, including quercetin, wogonin, baicalein, kaempferol, and oroxylin A.. Our research firstly revealed the basic pharmacological effects and relevant mechanisms of the HLJDD in the treatment of T2DM and its complications. The prediction results might facilitate the development of HLJDD or its active compounds as alternative therapy for T2DM. However, more pharmacological experiments should be performed for verification.

Topics: Apoptosis; China; Cluster Analysis; Diabetes Mellitus, Type 2; Drugs, Chinese Herbal; Flavanones; Flavonoids; Gene Expression Profiling; Humans; Inflammation; Interleukin-6; Kaempferols; Medicine, Chinese Traditional; Molecular Docking Simulation; Protein Interaction Mapping; Proto-Oncogene Proteins c-akt; Quercetin

Screening active components from Chinese traditional medicine is an effective approach to discover new drugs or active structures. Cell membrane chromatography (CMC), developed rapidly because of its high sensitivity and effectiveness, has achieved a wide application in screening active components on pathological cells or tissues. However, it is hard to clarify the selectivity between pathological and normal tissues through simply using pathological cells. In this study, a novel comparative two-dimensional (2D) cell membrane chromatography system was established. Briefly, hepatic carcinoma HepG2 CMC columns and normal hepatic L02 CMC columns were simultaneously loaded to screen potential selective antitumor components from Scutellariae Radix by comparing the retention behaviors on two kinds of cells. Totally 13 components in Scutellariae Radix retained on both HepG2/ CMC and L02/ CMC columns. Among them, three components, oroxylin A, wogonin and chrysin, were screened out to perform stronger affinity on HepG2 columns, and in further cell proliferation assay, IC

Topics: Antineoplastic Agents, Phytogenic; Carcinoma, Hepatocellular; Cell Membrane; Cell Proliferation; Chromatography, High Pressure Liquid; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; Flavanones; Flavonoids; Hep G2 Cells; Humans; Inhibitory Concentration 50; Liver Neoplasms; Mass Spectrometry; Scutellaria baicalensis

Studies on the efficacy evaluation of UDP-glucuronosyltransferases (UGTs) substrates often ignore the existence of active metabolites. However, the present study aims to establish an in-vitro Phase II metabolism system to predict their pharmacological effects after metabolism. Rat liver microsomes (RLMs) encapsulated in a F127'-Acr-Bis (FAB) hydrogel were placed in the incubation system. Baicalein (BA) was chosen as a model drug and the metabolic activity was investigated by quantitating the metabolite Baicalin (BG). The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to measure the cell viability in Traditional cell culture system (TCCS) and Microsome-hydrogel added to cell culture system for Phase II metabolism (MHCCS-II). Finally, MHCCS-II was applied to predict the metabolic effects of Oroxylin A (OA) and Wogonin (W). Compared to TCCS group, for HepG2 and MCF-7 cells, BA in MHCCS-II led to lower survival ratios of cells (P < 0.05), while for PC12 cells it led to higher survival ratios of cells (P < 0.01). For HepG2 cells, OA and W showed obviously enhanced tumor inhibition after metabolism with the IC

Topics: Animals; Antineoplastic Agents; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Flavanones; Flavonoids; Glucuronosyltransferase; Humans; Hydrogels; Metabolic Detoxication, Phase II; Microsomes, Liver; Molecular Structure; Rats; Structure-Activity Relationship; Tumor Cells, Cultured

Scutellaria root, the root of Scutellaria baicalensis Georgi, is a crude drug used for inflammatory diseases. In our previous report, the combination of flavonoids contained in Scutellaria root have been found to inhibit PGE

Topics: Drugs, Chinese Herbal; Flavanones; Flavonoids; Humans; NF-kappa B; Scutellaria; Scutellaria baicalensis; Transfection

Scutellariae Radix (SR) and its bioactive flavones elicit a variety of effects in the brain. However, the brain uptake of individual SR flavones and its relationship to the elicited effects after SR administration remain unknown. Moreover, previous studies seldom measured pharmacokinetic and pharmacodynamic outcomes simultaneously. In the current study, the brain uptake of six major SR flavones and the anxiolytic behavior following oral administration of a SR extract at two clinically relevant doses (600 and 1200 mg/kg twice daily) were simultaneously investigated in mice (n = 18 per group). Brain and plasma concentrations of the flavones were measured by LC-MS/MS, while the anxiolytic effect was evaluated using the elevated plus maze. To further investigate the mechanism behind the differential brain uptake of the six SR flavones, these flavones were separately administered to mice at an equivalent molar oral dose (n = 6). The brain tissue bindings of the SR flavones were also measured with the in vitro brain slice method. Our results indicated that all six SR flavones including three aglycons (baicalein, wogonin, and oroxylin A) and three glucuronides (baicalin, wogonoside, and oroxyloside) could pass through the blood-brain barrier, with brain concentrations ranging from 7.9 to 224.0 pmol/g. It provided novel evidence that oroxylin A had the highest brain uptake among the six SR flavones regardless of its limited content in SR extract, in which 3.6-3.9% of the administered oroxylin A dose was present in the brain 6 h postdosing and with a brain-to-plasma ratio of 0.42-0.46. Although SR extract contains flavones that are positive modulators of the benzodiazepine binding site of GABA

Topics: Animals; Anti-Anxiety Agents; Brain; Flavanones; Flavones; Flavonoids; Glucosides; Male; Maze Learning; Mice; Mice, Inbred ICR; Scutellaria baicalensis; Tandem Mass Spectrometry

Baicalein, wogonin, and oroxylin A are three major hydrophobic components in the extract of Radix scutellariae with wide spectrum of pharmacological applications. The purpose of this study was to enhance the solubility, dissolution rate and stability of baicalein, wogonin and oroxylin A by solid dispersion (SD) technique. SD of the extract with various polymers was prepared to select the best carrier. Solubility study, chemical stability study and dissolution study were performed to characterize the SD. The solubility of all three components, after forming solid dispersion with povidone K-30 (PVP K-30) was significantly increased in pH 6.8 medium at room temperature. Stability study conducted for 80days elucidated that the SD in powder state was fairly stable without the aid of Vitamin C (VC). VC was required as antioxidant to impart stability to baicalein in aqueous medium. The dissolution test of the SD of three components, admixed with VC at the weight ratio of 1:6 (Radix scutellariae extract: VC, w/w) exhibited faster dissolution rate with 100% release of all components. Pharmacokinetic study of baicalein solid dispersion revealed that AUC and C

Topics: Drug Stability; Flavanones; Flavonoids; Scutellaria; Solubility

Topics: Animals; Chromatography, High Pressure Liquid; Cyclooctanes; Drugs, Chinese Herbal; Emodin; Female; Flavanones; Flavonoids; Glucosides; Lignans; Limit of Detection; Linear Models; Male; Monoterpenes; Polycyclic Compounds; Rats; Rats, Wistar; Reproducibility of Results; Tandem Mass Spectrometry

As an important herbaceous plant, Scutellaria baicalensis Georgi (Chinese skullcap) is geographically widespread and commonly used throughout the world. In the Chinese medicine market, S. baicalensis has been divided into two primary types, "Ku Qin" (WXR) and "Tiao Qin" (TST). Moreover, TST is also divided into different grades according to the diameter of roots. To explore the distribution patterns of the contents of five biologically activate ingredients (FBAI), we used six-year-old cultivated S. baicalensis and analyzed its growth characteristics as well as the quality difference among different types and diameters in roots. Throughout the entire root, we discovered that contents of the FBAI all initially increased and subsequently decreased from the top to the bottom of the roots. The baicalin content of WXR was less than that of TST. On the contrary, the contents of baicalein, wogonin, and oroxylin A in WXR were up to about two times higher than that in TST. We also found that the 0 to 40 cm part of the S. baicalensis root possessed about 87% of the root biomass and about 92% of the contents of the active ingredients.

Topics: Drugs, Chinese Herbal; Flavanones; Flavonoids; Plant Roots; Scutellaria baicalensis

A new method consisting of orthogonal projection to latent structure (OPLS) and modified principal component analysis (PCA) was applied to a screening of bioactive compounds from natural products. In this report, extracts of 52 Scutellaria Root (the root of Scutellaria baicalensis Georgi) samples were analyzed by high-performance liquid chromatography (HPLC), and their inhibitory activities towards prostaglandin E2 (PGE2) production in a murine macrophage-like cell line J774.1 were examined. Wogonin and oroxylin A were predicted to be strong inhibitors of PGE2 production by OPLS analysis of the data. However, 6-methoxywogonin, which has been reported to have inhibitory activity, was omitted. Modified PCA was then applied to these data as a filter to exclude compounds less relevant to the activity, and OPLS analysis was applied to the modified data. As a result, this method predicted wogonin, oroxylin A and 6-methoxywogonin to be strong inhibitors of PGE2 production without any prior knowledge. The predictions by the OPLS combined with PCA method of PGE2 production inhibitory activities of 52 samples showed good agreement with the actual data. This method is simple and effective and can be used in screening of bioactive natural compounds without any prior knowledge.

Topics: Animals; Cell Line; Chromatography, High Pressure Liquid; Dinoprostone; Drug Evaluation, Preclinical; Flavanones; Flavonoids; Mice; Plant Extracts; Plant Roots; Principal Component Analysis; Scutellaria baicalensis

Mefenamic acid (MEF) and the dried root of Scutellaria baicalensis Georgi (Radix Scutellariae, RS) share a high possibility of combined medication to treat inflammation.. The present study investigates the impact of MEF on absorption/disposition of three major components in RS (baicalein, B; wogonin, W; oroxylin A, OA) and further pharmacological changes.. The apparent permeability (P(app)) and percentage of metabolism of B, W and OA at 10 μΜ were measured at the absence/presence of MEF (100 μΜ) in the Caco-2 cell monolayer model. A modified whole blood assay was employed to quantify prostaglandin E₂ (PGE₂) 4, 6 and 8 h post-oral administration with water suspension of MEF at 40 mg/kg and RS at 200 mg/kg.. In the presence of MEF, Papp of B, W and OA were increased from 1.69 ± 0.89 × 10⁻⁶, 1.57 ± 0.10 × 10⁻⁶ and 3.09 ± 0.70 × 10⁻⁶ cm/sec to 5.24 ± 0.27 × 10⁻⁶, 6.08 ± 0.19 × 10⁻⁶ and 4.13 ± 0.38 × 10⁻⁶, whereas their percentage of metabolism was decreased from 72.75 ± 2.44%, 73.27 ± 3.25% and 89.84 ± 2.99% to 21.11 ± 0.69%, 17.90 ± 5.55% and 45.44 ± 3.38%. PGE2 level was much lower in the co-administration group (49.04 ± 2.03 pg/ml) than in the MEF group (73.13 ± 3.03 pg/ml) or RS group (494.37 ± 11.75 pg/ml) 4 h post MEF dosing, suggesting a synergic effect.. Co-administration of MEF and RS could induce potential alterations in their pharmacokinetic profiles and anti-inflammatory effects.

Topics: Animals; Anti-Inflammatory Agents; Caco-2 Cells; Drug Interactions; Drug Synergism; Flavanones; Flavonoids; Humans; Intestinal Absorption; Male; Mefenamic Acid; Plant Extracts; Plant Roots; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Time Factors

1. This study elucidated the species differences between rats and humans in the inhibitory potential of drugs against sulfation and glucuronidation, and whether such differences depend on the inhibition parameter adopted. 2. With 14 non-steroidal anti-inflammatory drugs (NSAIDs) as model inhibitors and three flavanoids baicalein, wogonin and oroxylin A as model substrates, three common inhibition parameters percentage of control, IC50 and Ki were determined in rat liver cytosols (RLCs), human liver cytosols (HLCs), rat liver microsomes (RLMs) and human liver microsomes (HLMs). The closeness of the inhibition parameters from rat liver preparations to that from human liver preparations was analyzed by geometric mean fold error (GMFE) and statistical comparisons. 3. The percentage of control in RLC/RLM was not significantly different from that in HLC/HLM, with a GMFE of 0.85 (RLC-HLC) and 1.03 (RLM-HLM); whereas the IC50 and Ki in RLC/RLM were significantly different from that in HLC/HLM. The trend of difference was consistent between IC50 and Ki, where these parameters in RLC and RLM underestimated (GMFE <0.5) and overestimated (GMFE >2) that in HLC and HLM, respectively. 4. In conclusion, the inhibitory potentials of NSAIDs against sulfation and glucuronidation in rats and humans were different and depended on the adopted inhibition parameters.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cytosol; Dose-Response Relationship, Drug; Flavanones; Flavonoids; Humans; Inhibitory Concentration 50; Liver; Microsomes, Liver; Rats; Species Specificity

This study aims to identify and quantify the six major bioactive flavones of the traditional Chinese medicine Scutellariae Radix (RS), including baicalein, baicalin, wogonin, wogonoside, oroxylin A and oroxyloside in rat after oral administration of a standardized RS extract. A novel, sensitive and selective method for simultaneous determination of these six analytes in rat brain and plasma using solid phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC/MS/MS) was developed and fully validated. The lower limits of quantification (LLOQs) for the six RS flavones in brain tissue were 0.02nmol/g. The LLOQs in plasma were 0.005nmol/ml for B, W and OA, 0.025nmol/ml for WG and OAG, and 0.1875nmol/ml for BG. The current study provides novel evidence of the presence of all the tested RS flavones and an isoform of BG (BG', probably baicalein-6-O-glucuronide) in the rat brain after oral administration of RS extract, suggesting their ability to permeate through the blood-brain barrier. The method was also successfully applied to the pharmacokinetic study of all these analytes in plasma after oral administration of RS extract (300mg/kg) to Sprague-Dawley rats. The developed assay method provides a useful tool for both preclinical and clinical investigations on the disposition of RS flavones in brain and plasma.

Topics: Animals; Brain; Chromatography, Liquid; Flavanones; Flavones; Flavonoids; Glucosides; Glucuronides; Male; Rats; Solid Phase Extraction; Tandem Mass Spectrometry

The pharmacodynamic (PD) and pharmacokinetic (PK) properties of Huangqin Tang (HQT) were investigated in yeast-induced febrile rats. Blood sample and rectal temperature data of the rats were collected at different times after single oral administration of HQT at 20 g x kg(-1). The plasma concentrations of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid were quantified by a sensitive liquid chromatography-tandem mass spectrometric (LC-MS) method. The blood concentrations of PGE2, 1L-1β and TNF-α were detected by radioimmunoassay (RIA). All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The potential relationship between the mean concentration of eight constituents and the antifebrile efficacy was investigated by calculating Pearson correlation coefficients. It was found that HQT had significant antifebrile efficacy in yeast-induced febrile rats, but had no effect to normal rats. The antifebrile effect of HQT can be attributed to the inhibition of PGE2, 1L-1β and TNF-α. The constituents (baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid) in febrile rats had delayed absorption and elimination, a longer residence time in the body, and higher C(max) and AUC than those in normal rats. Febrile condition could affect the pharmacokinetic behaviour of HQT in vivo; the flavonoids with the same backbone showed the similar fate in the body; baicalein and wogonin had a strong positive correlation (R > 0.66, P ≤ 0.02) with the antifebrile efficacy determined. Together, these constituents demonstrated different pharmacokinetic properties in the febrile body.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, Liquid; Dinoprostone; Drugs, Chinese Herbal; Fever; Flavanones; Flavonoids; Glucosides; Interleukin-1beta; Mass Spectrometry; Monoterpenes; Rats; Tumor Necrosis Factor-alpha

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.

Topics: Administration, Oral; Animals; Chromatography, Liquid; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Male; Monoterpenes; Pentacyclic Triterpenes; Rats; Rats, Wistar; Spectrometry, Mass, Electrospray Ionization

A number of oroxylin A analogs were prepared and evaluated for their inhibitory activities against iNOS-mediated nitric oxide (NO) production from LPS-stimulated BV2 cells. The analogs were synthesized from purchased 2'-hydroxy-4,5,6-trimethoxyacetophenone and aldehydes in 3 steps. Among the tested compounds, several analogs (3b, 3c, 3d, 3f) exhibited strong inhibitory activities. Especially, the analog with 4-nitrophenyl group (3b) showed stronger inhibitory activity (IC(50)=4.73 μM) than that of wogonin (IC(50)=7.80 μM).

Topics: Acetophenones; Aldehydes; Animals; Anti-Anxiety Agents; Cell Line, Transformed; Flavanones; Flavonoids; Lipopolysaccharides; Macrophages; Mice; Microglia; Nitric Oxide; Nitric Oxide Synthase Type II; Nitrophenols

Radix Scutellariae is a commonly used herbal medicine. Baicalein, wogonin, and oroxylin A are three major bioactive flavones in Radix Scutellariae and share similar chemical structures. The intestinal absorption and disposition of baicalein have been systematically investigated by our group before. In this study, the intestinal absorption and disposition of wogonin and oroxylin A were further explored and compared with the profiles of baicalein to find potential structure-activity relationship. Absorptive models including Caco-2 cell monolayer model and rat in situ single-pass intestinal perfusion model as well as in vitro enzymatic kinetic study were employed in the current study. The absorption of baicalein, wogonin, and oroxylin A were favorable with wogonin showing the highest permeability based on two absorptive models. However, three flavones underwent a fast and extensive phase II metabolism. The intestinal metabolism of three flavones exhibited species difference between human and rat. Oroxylin A demonstrated the highest intrinsic clearance of glucuronidation among three flavones. The multidrug resistance proteins might be involved in the efflux of their intracellularly formed conjugated metabolites. The pathway of intestinal absorption and disposition of B, W, and OA was similar. However, the extent of permeability and metabolism was different among three flavones which might be due to the number and position of the hydroxyl group.

Topics: Animals; Caco-2 Cells; Flavanones; Flavonoids; Glucuronides; Humans; Intestinal Absorption; Male; Permeability; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Species Specificity; Structure-Activity Relationship

A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of baicalin, wogonoside, baicalein, wogonin, oroxylin A and chrysin in rat plasma, using naringin as an internal standard. After acidifying with HCl, plasma samples were pretreated by liquid-liquid extraction with acetone. Chromatographic separation was accomplished on a Hypersil Gold-C(18) analytical column (2.1×150 mm, 5 μm) utilizing a gradient elution profile and a mobile phase consisting of (A) 0.1% formic acid in water and (B) acetonitrile. Detection was performed by multiple reaction monitoring (MRM) mode using electrospray ionization in the positive ion mode. All analytes showed good linearity over the investigated concentration range (r>0.9900). The lower limit of quantification was 0.5 ng/ml for baicalin, wogonoside, wogonin and oroxylin A, and 1.0 ng/ml for baicalein and chrysin. Intra-day and inter-day precisions (RSD%) were less than 15% and accuracy (RE%) ranged from -6.7% to 5.8%. The validated method was successfully applied to investigate the pharmacokinetics of the major flavonoids of Radix scutellariae extract after oral administration to rats.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Half-Life; Limit of Detection; Male; Metabolic Clearance Rate; Plant Extracts; Rats; Rats, Wistar; Reference Standards; Reproducibility of Results; Scutellaria baicalensis; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Flavonoid reference standards were targeted-prepared from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis. With HPLC-MS analysis of Scutellariae Radix, 19 flavonoid components were identified by analyzing and comparing their retention times, ultraviolet spectra, and mass spectrometry data with literature. The separation and purification protocols of all targeted flavonoid reference standards were optimally designed according to the results of HPLC-MS analysis and related literature. The ethanol extract of Scutellariae Radix was suspended in water and extracted with petroleum ether, ethyl acetate, and n-butanol successively. The ethyl acetate extract and n-butanol extract were separately subjected to primary separation by low pressure reverse phase preparative chromatography. Then the fractions containing targeted compounds were further purified by low pressure reverse and normal phases preparative chromatography. Finally, baicalin and wogonoside reference standards were obtained from n-butanol extract; baicaelin, wogonin, and oroxylin A reference standards were obtained from ethyl acetate extract. The structures of the 5 reference standards were identified by mass spectrometry (MS) and 1H nuclear magnetic resonance (1H NMR) spectroscopy. The HPLC analytical results showed that the purities of the 5 reference standards were all above 98%. It is demonstrated that the rapid targeted-preparation method under the guidance of the HPLC-MS analysis is applicable for the isolation and preparation of chemical components in traditional Chinese medicines.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Mass Spectrometry; Reference Standards; Scutellaria baicalensis

The current study aims to identify and quantify three flavones (baicalein, wogonin and oroxylin A) and their major metabolites (baicalin, wogonoside and oroxylin A-7-O-glucuronide) in rat plasma after oral administration of Radix scutellariae product. A simple HPLC/UV method has been developed to simultaneously determine the three flavones and their major metabolites in rat plasma. The chromatographic separation of the six analytes was achieved by a Thermo C(18) column with linear gradient elution of a mobile phase containing acetonitrile and 20mM sodium dihydrogen phosphate buffer (pH 4.6). All the tested analytes were detected by PDA detector at a wavelength of 320nm. The intra-day and inter-day precision for the current assay of the six analytes was within the range of -2.23% to 15.13% and -10.83% to 6.42%, respectively. All the studied analytes could be efficiently extracted from the rat plasma using HLB cartridge with extraction recoveries above 70% and were stable under different storage conditions. The developed assay method was successfully applied to the pharmacokinetic study of baicalin, wogonoside after oral administration of a commercially available Radix scutellariae containing capsule at a dose of 3.2g/kg to Sprague-Dawley rats. In addition to wogonoside, a new metabolite of wogonin has been identified using LC/MS/MS for the first time.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Drug Stability; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucuronides; Limit of Detection; Male; Molecular Structure; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Scutellaria baicalensis; Solid Phase Extraction; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Glucuronidation via UDP-glucuronosyltransferases (or UGTs) is a major metabolic pathway. The purposes of this study are to determine the UGT-isoform-specific metabolic fingerprint (or GSMF) of wogonin and oroxylin A, and to use isoform-specific metabolism rates and kinetics to determine and describe their glucuronidation behaviors in tissue microsomes.. In vitro glucuronidation rates and profiles were measured using expressed UGTs and human intestinal and liver microsomes.. GSMF experiments indicated that both flavonoids were metabolized mainly by UGT1As, with major contributions from UGT1A3 and UGT1A7-1A10. Isoform-specific metabolism showed that kinetic profiles obtained using expressed UGT1A3 and UGT1A7-1A10 could fit to known kinetic models. Glucuronidation of both flavonoids in human intestinal and liver microsomes followed simple Michaelis-Menten kinetics. A comparison of the kinetic parameters and profiles suggests that UGT1A9 is likely the main isoform responsible for liver metabolism. In contrast, a combination of UGT1As with a major contribution from UGT1A10 contributed to their intestinal metabolism. Correlation studies clearly showed that UGT isoform-specific metabolism could describe their metabolism rates and profiles in human liver and intestinal microsomes.. GSMF and isoform-specific metabolism profiles can determine and describe glucuronidation rates and profiles in human tissue microsomes.

Topics: Flavanones; Flavonoids; Gas Chromatography-Mass Spectrometry; Glucuronides; Glucuronosyltransferase; Humans; Intestinal Mucosa; Kinetics; Microsomes; Microsomes, Liver; Monosaccharide Transport Proteins; Protein Isoforms

Two new dihydropyranocoumarins, scuteflorins A (1) and B (2), together with the known compounds decursin (3), chrysin (4), oroxylin A (5), wogonin (6), 5,7-dihydroxy-8,2'-dimethoxyflavone, dihydrochrysin, dihydrooroxylin A, lupenol, scutellaric acid, pomolic acid, ursolic acid, beta-sitosterol, daucosterol, and palmitic acid, were isolated from the aerial parts of Scutellaria lateriflora, commonly used as a dietary supplement. The structures of 1 and 2 were established by means of 1D and 2D NMR spectra as well as HRMS data. The absolute configuration of coumarins 1 and 2 was determined by comparison of experimental and theoretical calculated CD spectra. The cytotoxicity and antioxidant effects of the methanol extract of this plant and some of the constituent flavonoids were evaluated in vitro.

Topics: Algorithms; Animals; Chlorocebus aethiops; Drug Screening Assays, Antitumor; Flavanones; Flavonoids; HL-60 Cells; Humans; Nuclear Magnetic Resonance, Biomolecular; Pyranocoumarins; Vero Cells

To study the ethyl acetate soluble constituents from the water extractive of Huanglian Jiedutang decoction, which are composed of Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri and Fructus Gardeniae, and provide substances foundation for its pharmacokinetic and pharmacodynamic investigation.. The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by NMR and MS techniques.. Thirty-five compounds were isolated, among which twenty compounds have been identified as beta-sitosterol (1), oroxylin A (2), wogonin (3), ursolic acid (4), skullcapflavone I (5), tenaxin I (6), skullcapflavone II (7), limonin (8), 5, 2'-dihydroxy-6, 7, 8, 3'-tetramethoxyflavone (9), chrysin (12), baicalein (17), tenaxin II (19), 5, 7, 2'-trihydroxy-6, 8-dimethoxyflavone (21), shihulimonin A (22), 6, 2'-dihydroxy-5, 7, 8, 6'-tetramethoxyflavone (26), viscidulin II (28), 5, 7, 4'-trihydroxy-8-methoxyflavone (29), 5, 7, 2', 6'-tetrahydroxyflavone (30), wogonin-7-O-beta-D-glucuronide methyl ester (31) and daucosterol (34).. On the basis of reported results of the chemical constituents of Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri and Fructus Gardeniae, it was estimated that all flavonoid compounds rised from the Radix Scutellariae, and compounds 8 and 22 rised from Cortex Phellodendri. Compound 22 was identified in the Cortex Phellodendri for the first time.

Topics: Acetates; Drugs, Chinese Herbal; Flavanones; Flavones; Flavonoids; Limonins; Magnetic Resonance Spectroscopy; Sitosterols; Spectrometry, Mass, Electrospray Ionization; Triterpenes; Ursolic Acid

The pharmacokinetics of active components such as baicalein, wogonin and oroxylin A were evaluated after oral administration of a purified extract of Scutellaria baicalensis GEORGI (PF-2405) containing the high contents of baicalein, wogonin and oroxylin A to rats. Following oral administration of PF-2405 at 10, 20 and 40 mg/kg dose (equivalent to 4.5, 9.0 and 18 mg/kg baicalein), a major constituent baicalein and its active metabolite baicalin showed dose-linear pharmacokinetics as evidenced by unaltered dose-normalized AUC, dose-normalized Cmax, Ae(0-30h) and GI(30h) values. Following oral administration of PF-2405 at three doses (equivalent to 0.4, 0.8 and 1.6 mg/kg wogonin), dose-normalized Cmax and dose-normalized AUC were comparable between the 20 and 40 mg/kg PF2405 doses, but plasma concentrations of wogonin at 10 mg/kg of PF-2405 were not measurable as they were below limit of quantitation (LOQ; 18 pmol/mL). Following oral administration of PF-2405 at the three doses (equivalent to 1.5, 3.0 and 6.0 mg/kg oroxylin A), the concentrations of oroxylin A in plasma, urine and gastrointestine samples were below the assay LOQ (18 pmol/mL). Significant differences in AUCs, Ae(0-30h) and GI(30h) values for baicalein and baicalin were observed after oral administration of pure baicalein (18 mg/kg) and PF-2405 (40 mg/kg). The increases in AUCs of baicalein and baicalin after oral administration of PF-2405 may have been due to the significant decrease in GO(30h) values for baicalein.

Topics: Administration, Oral; Animals; Chromatography, Liquid; Flavanones; Flavonoids; Gastrointestinal Tract; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Tandem Mass Spectrometry

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of baicalein, baicalin, oroxylin A and wogonin, Scutellaria baicalensis active components in rat plasma was developed. After liquid-liquid extraction with 2-(3,4-dimethoxy-phenyl)-5,7-dihydroxy-chromen-4-one as internal standard, baicalein, baicalin, oroxylin A and wogonin were eluted from an Atlantis C(18) column within 7 min with isocratic mobile phase consisting of methanol and 0.1% formic acid (60:40, v/v). The analytes were detected using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode. The standard curves were linear (r=1.000) over the concentration ranges of 5-500 ng/ml for baicalein, wogonin and oroxylin A and 5-5000 ng/ml for baicalin. The coefficients of variation and relative errors of baicalein, wogonin, oroxylin A and baicalin for intra- and inter-assay at three or four quality control (QC) levels were 0.8-6.1% and -4.0 to 5.8%, respectively. The lower limits of quantification for baicalein, wogonin, oroxylin A and baicalin were 5ng/ml using 50 microl of plasma sample. This method was successfully applied to the pharmacokinetic study of baicalein, baicalin, wogonin and oroxylin A after an intravenous administration of Scutellariae radix extract to male Sprague-Dawley rats.

Topics: Animals; Chromatography, Liquid; Flavanones; Flavonoids; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Scutellaria; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

The active ingredients of 'golden root' of Scutellaria baicalensis GEORGI (Huang-Qin), a valuable traditional Chinese medicine, are polyhydroxyflavonoids, namely baicalein, oroxylin A and wogonin. With the objective of overcoming their poor solubility and to investigate their structure and activity relationships, baicaleinyl 7-O-sulfate was prepared, and extensive comparative antioxidative and anti-inflammatory tests were conducted. All the polyhydroxyflavonoids exhibited significant antioxidative and free-radical scavenging activities. In respect of their nitric oxide (NO) inhibition, wogonin was superior to all the other flavonoids, while oroxylin A was most potent in the inhibition of lipid peroxidation. Wogonin proved to be the most potent (82.9% inhibition, p<0.05) in its anti-inflammatory activity against carrageenan-induced rat hind paw edema. There was a correlation between the in-vivo anti-inflammatory activity and the in-vitro antioxidative activities.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Drugs, Chinese Herbal; Edema; Flavanones; Flavonoids; Free Radical Scavengers; Lipid Peroxidation; Nitric Oxide; Phytotherapy; Rats; Scutellaria baicalensis; Solubility; Structure-Activity Relationship

The medicinal plant Scutellaria baicalensis Georgi has been used widely in traditional Chinese medicine for anti-inflammation, anticancer, antiviral and antibacterial infections, reducing the total cholesterol level and decreasing blood pressures. A high-speed counter-current chromatography (HSCCC) method was developed for the preparative separation and purification of three bioactive flavonoids, namely, baicalein, wogonin and oroxylin A, from S. baicalensis Georgi. Preparative HSCCC with a two-phase solvent system composed of n-hexane-ethyl acetate-n-butanol-water (1:1:8:10, v/v/v/v) was successfully performed by increasing the flow-rate of the mobile phase stepwise from 1.0 to 2.0 ml min(-1) after 4 h. The components purified and collected were analyzed by high-performance liquid chromatography. The method yielded 144.8 mg of baicalein at 95.7% purity, 50.2 mg of wogonin at 98.5% purity, and 12.4 mg of oroxylin A at 93.2% purity from 500 mg of the crude extract in a one-step separation. The recoveries of baicalein, wogonin and oroxylin A were 92.7%, 91.6% and 92.5%, respectively.

Topics: Chromatography, High Pressure Liquid; Countercurrent Distribution; Flavanones; Flavonoids; Scutellaria baicalensis

Inhibitory mechanism of the water extract of Scutellariae Radix on prostaglandin E2 (PGE2) release was examined in C6 rat glioma cells. Scutellariae Radix reduced a Ca2+ ionophore A23187-induced PGE2 release by inhibition of arachidonic acid (AA) liberation. Sho-saiko-to and San'o-shashin-to, which contain Scutellariae Radix, also inhibited PGE2 release. A23187 caused phosphorylation of mitrogen-activated protein kinase (MAPK), resulting in activation of cytosolic phospholipase A2 (cPLA2). Scutellariae Radix and baicalein inhibited the phosphorylation of MAPK. Baicalein, but not baicalin, inhibited A23187-induced PGE2 release. These results suggest that baicalein in Scutellariae Radix reduces AA liberation through the inhibition of the MAPK-cPLA2 pathway.

Topics: Amino Acid Sequence; Animals; Antineoplastic Agents; Arachidonic Acid; Calcimycin; Calcium-Calmodulin-Dependent Protein Kinases; Dinoprostone; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Glioma; Ionophores; Molecular Sequence Data; Phosphorylation; Prostaglandin Antagonists; Rats; Tumor Cells, Cultured