ABSTRACT
Two new chromones named cnidimol G (1) and cnidimol H (2), one new coumarin, 7-methoxy-8-(3-methoxy-3-methyl-2-oxobutyl)coumarin (3), and twenty known compounds were isolated from MeOH extract of the fruit of Cnidium monnieri (L.) Cusson. The structures of compounds were elucidated by extensive spectroscopic analyses including 1 D and 2 D NMR, HRESIMS, IR and UV. Anti-inflammatory activity of the selected isolated compounds were evaluated. Compounds 1 and 8 exhibited inhibitory activities against nitric oxide production.
Subject(s)
Cnidium , Fruit , Cnidium/chemistry , Fruit/chemistry , Chromones/pharmacology , Chromones/analysis , Plant Extracts/chemistry , Coumarins/chemistryABSTRACT
Qu-feng-sheng-shi Granules (QFSSG), a common prescription for the treatment of chronic inflammation and allergic rhinitis, is widely used in the clinic as a traditional Chinese medicine. Chemical analysis and quality control studies of this formulation are relatively limited compared with pharmacological studies. In this study, a high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q/TOF-MSn ) was used to identify the components in this prescription. Next, to quantify six major compounds, an HPLC-UV method was developed and validated. The results showed that 53 compounds were identified based on the MSn data, retention time and previous reports, including 17 coumarins, 14 lignans, 10 chromones, nine phenylethanoid glycosides and three other compounds, were identified or tentatively assigned. Contents of six major bioactive compounds (4'-O-ß-glucopyranosyl-5-O-methylvisamminol, Prim-O-glucosylcimifugin, forsythin, magnolin, imperatorin, isoimperatorin) could be determined by HPLC simultaneously. In addition, the potential anti-inflammatory activity of six major compounds was determined too, and we found that four compounds (4'-O-ß-glucopyranosyl-5-O-methylvisamminol, Prim-O-glucosylcimifugin, forsythin, imperatorin) have a potent nitric oxide inhibitory effect. In conclusion, this work provided comprehensive information on the quality control of QFSSG and evaluated the potential biological activity of the main components in QFSSG, which can contribute to understanding and using it more scientifically.
Subject(s)
Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal , Spectrometry, Mass, Electrospray Ionization/methods , Animals , Cell Survival/drug effects , Chromones/analysis , Chromones/chemistry , Coumarins/analysis , Coumarins/chemistry , Drugs, Chinese Herbal/analysis , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Glycosides/analysis , Glycosides/chemistry , Lignans/analysis , Lignans/chemistry , Limit of Detection , Linear Models , Mice , RAW 264.7 Cells , Reproducibility of Results , Spectrophotometry, UltravioletABSTRACT
Agarwood, a species of resinous heartwood, is a precious medicinal plant and a type of rare natural spice, which is widely used in medicine, cosmetics, religious activities, and other fields. In this study, agarwood samples from eight different regions across four countries were analyzed by comprehensive two-dimensional gas chromatography-quadrupole time-of-flight mass spectrometry. A total of 232 species were identified (the match factors of these compounds were above 750). The main compounds of agarwood are oxygenated sesquiterpenes and chromones. The compositions of India1 and Malaysia2 were significantly different from those of other samples, which might be attributed to the different production processes of agarwood. For further investigation, factor analysis was conducted for six agarwood samples. The results showed that the data classification possessed a regional characteristic; according to the retention time and relative content, characteristic compositions were determined by factor scores. Finally, the differences of characteristic compositions were simply analyzed, and the reasons were speculated.
Subject(s)
Chromones/analysis , Sesquiterpenes/analysis , Thymelaeaceae/chemistry , Gas Chromatography-Mass Spectrometry/instrumentationABSTRACT
Saposhnikoviae Radix (SR) is a commonly used crude drug that is obtained from the root and rhizome of Saposhnikovia divaricata which is distributed throughout China, Korea, Mongolia, and Russia. To evaluate the quality of Mongolian S. divaricata, metabolomic profiling of 43 plant specimens from different regions of Mongolia, as well as 8 SR samples and 2 plant specimens from China, were conducted by liquid chromatography-ion-trap-time-of-flight-mass spectrometer (LC-IT-TOF-MS). LC-MS profiles of the specimens showed uniformity and 30 compounds were tentatively identified, including 13 chromones and 17 coumarins. Among them, 16 compounds were isolated and unambiguously verified by comparing them with the spectroscopic data of standard compounds. Orthogonal partial least squares-discriminant analysis (OPLS-DA) based on LC-MS data from 7 Mongolian specimens and 8 Chinese SR samples as well as 2 plant specimens revealed that these 2 groups were clearly distinguishable and that Mongolian specimens were characterized by an abundance of prim-O-glucosylcimifugin (1). Moreover, the OPLS-DA of the Mongolian specimens showed that they can be discriminated by their growing regions based on the content of 8 chromones. The total content of dihydrofurochromones 1-3 was relatively higher in the specimens from Khalkhgol in the far eastern part of Mongolia, while contents of 10, 11, 15, and 16 were higher in those from Holonbuir in the eastern part. Based on this research, the roots of S. divaricata from Mongolia have potential as a new resource of SR in Kampo medicine.
Subject(s)
Apiaceae/chemistry , Chromones/analysis , Chromones/chemistry , Coumarins/chemistry , Monosaccharides/chemistry , Xanthenes/chemistry , China , Chromatography, High Pressure Liquid/methods , Chromatography, Liquid , Drugs, Chinese Herbal/chemistry , Medicine, Kampo , Mongolia , Plant Roots/chemistry , Rhizome/chemistry , Tandem Mass Spectrometry/methodsABSTRACT
Agarwood, which is used as medicine and incense, contains sesquiterpenes and chromones. Agarotetrol is a chromone derivative found in high concentrations in the water-extract fraction of agarwood and thus may be present in pharmaceutical products made from decoctions of agarwood. Agarotetrol has been reported to be present at the early stages of cell death in calli. We therefore examined the presence of agarotetrol in medical- and incense-grade agarwood, in agarwood-source plants lacking resin deposits, and in artificially made agarwood. Agarotetrol appeared as a large peak in the HPLC chromatograms of all samples of medical-grade and artificially made agarwood, and in most incense-grade agarwood samples. In contrast, agarwood samples lacking resin deposits did not contain agarotetrol. These results show that agarotetrol is characteristic of resin formation. Agarotetrol was also detected in decoctions of agarwood. A newly developed TLC method for the detection of agarotetrol in agarwood is described.
Subject(s)
Chromones/analysis , Chromones/chemistry , Resins, Plant/analysis , Thymelaeaceae/chemistry , Chromatography, High Pressure Liquid , Plant Extracts , Resins, Plant/chemistry , Sesquiterpenes/metabolismABSTRACT
Limonoids, quinolone alkaloids and chromones have been reported as constituents of Dictyoloma vandellianum Adr. Juss. (Rutaceae). Although those compounds are known for their biological activities, only the anti-inflammatory activity of chromones isolated from the underground parts has been evaluated. There are no studies of the pharmacological properties of the aerial parts of D. vandellianum. The present study was carried out to determine the phytochemical profile and antinociceptive activity of the methanol extract, fractions and isolated compounds of leaves of D. vandellianum. The phytochemical profile was performed by HLPC-DAD-ESIMSn and pure substances obtained were characterized by MS and NMR spectroscopy. The antinociceptive activity was assessed using the formalin assay in mice, and the motor function in the rotarod test. ME and all the fractions obtained from ME produced antinociceptive effects. Among them, the ethyl ether fraction was the most active. Data from HPLC-DAD-ESIMSn showed that the ethyl ether fraction presented 42 compounds. The major compounds isolated from this fraction-gallic acid, methyl gallate and 1,2,6-tri-O-galloyl-ß-d-glucopyranose-were tested and produced antinociceptive effects. Gallic acid, methyl gallate and 1,2,6-tri-O-galloyl-ß-d-glucopyranose at antinociceptive doses did not affect the motor performance in mice in the rotarod test. This work is the first report of the occurrence of gallotanins in D. vandellianum. In addition, the pharmacological study showed that D. vandellianum leaves present antinociceptive activity, probably induced by gallic acid, methyl gallate and 1,2,6-tri-O-galloyl-ß-d-glucopyranose.
Subject(s)
Analgesics/chemistry , Plant Leaves/chemistry , Rutaceae/chemistry , Alkaloids/chemistry , Animals , Chromatography, High Pressure Liquid/methods , Chromatography, Liquid/methods , Chromones/analysis , Gas Chromatography-Mass Spectrometry/methods , Limonins/analysis , Male , Methanol/analysis , Mice , Phytochemicals/analysis , Plant Extracts/pharmacology , Plant Leaves/metabolism , Rutaceae/geneticsABSTRACT
BACKGROUND: Long-term exposure to aeroallergens such as house dust mite (HDM) could result in airway inflammation and airway remodeling, characteristic features of allergic asthma. Huangqi-Fangfeng (HF), an important "couplet medicines" of Yu-Ping-Feng-San (YPFS), mediates allergen-induced airway inflammation in mice, but its role in the airway remodeling is not known. PURPOSE: To evaluate the effects of HF on airway remodeling of allergic asthma in a murine model and to investigate the underlying mechanisms in vivo and in vitro. METHODS: The main components of HF were analyzed by HPLC. The HDM-induced asthma mice model was established to study the effects of HF on airway inflammation and airway remodeling in vivo. Enhanced pause (Penh) index value was used as an indicator of airway hyper-reactivity. Bronchoalveolar lavage fluid (BALF) was processed for differential cell counting and determination of cytokines production. The lungs were fixed in 4% paraformaldehyde for histological examination after staining with H&E, trichrome and IHC. Production of interleukin (IL)-4, IL-5, IL-13, and transforming growth factor beta-1 (TGF-ß1) in BALF and lung tissues, IgE in serum were measured by ELISAs. Expression of epithelial markers and mesenchymal markers were detected by immunohistochemistry and western blots. The effects of HF and its components on epithelial-mesenchymal transition (EMT) were detected in human bronchial epithelial cells (16HBE) treated with TGF-ß1 and HDM. RESULTS: The main components of Huangqi-Fangfeng detected by HPLC were Calycosin, Formononetin and Cimifugin. In HDM-induced allergic asthma mice model, respiratory exposure to HDM lead to airway hyperresponsiveness and thickening of the smooth muscle layer in the airway. TGF-ß1 levels increased in mice airways while epithelial cells lost expression of E-cadherin and gained expression of the mesenchymal proteins N-cadherin, α-SMA and collagen Ð. These changes were relieved by treatment with HF. Furthermore, restored epithelial markers expression treated with individual components were also detectable in 16HBE cells. CONCLUSION: These results demonstrated that Huangqi-Fangfeng protected against allergic airway remodeling through inhibiting epithelial-mesenchymal transition process in mice via regulating epithelial derived TGF-ß1.
Subject(s)
Airway Remodeling/drug effects , Anti-Asthmatic Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Epithelial-Mesenchymal Transition/drug effects , Animals , Anti-Asthmatic Agents/chemistry , Apiaceae/chemistry , Asthma/etiology , Asthma/pathology , Astragalus propinquus , Bronchi/cytology , Bronchoalveolar Lavage Fluid , Chromones/analysis , Disease Models, Animal , Drugs, Chinese Herbal/analysis , Drugs, Chinese Herbal/chemistry , Epithelial Cells/drug effects , Humans , Isoflavones/analysis , Lung/drug effects , Lung/pathology , Male , Mice, Inbred BALB C , Transforming Growth Factor beta1/metabolismABSTRACT
ãSaposhnikoviae Radix ("Boufu") is an important crude drug used in Kampo formulation. It is extracted from wild-type plants. However, recently, extraction has become difficult because of a decrease in wild-type plants. Therefore, cultivated plants account for the majority of the market, from which the crude drug is extracted. However, the cultivation techniques used are not sufficient to obtain the desirable extracts. In this study, we compared the contents of the extract and the quantitative values of characteristic constituents obtained from wild-type and cultivated plants, and found a remarkable difference. Therefore, it is considered that these indicators play an important role in the establishment of better cultivation technology.
Subject(s)
Apiaceae/chemistry , Culture Techniques/methods , Plant Extracts/chemistry , Plant Roots/chemistry , Chromatography, High Pressure Liquid , Chromones/analysis , Chromones/chemistry , Chromones/isolation & purification , Glucosides/analysis , Glucosides/chemistry , Glucosides/isolation & purification , Molecular Conformation , Monosaccharides/analysis , Monosaccharides/chemistry , Monosaccharides/isolation & purification , Xanthenes/analysis , Xanthenes/chemistry , Xanthenes/isolation & purificationABSTRACT
Saposhnikoviae Radix (SR), the dried root of Saposhnikovia divaricata (Turcz.) Schischk. (Umbelliferae), is commonly used as a traditional Chinese medicine. In this study, a rapid and accurate method was firstly, developed for the qualitative analysis of SR by high-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS/MS). A total of 45 compounds were identified or tentatively characterised, including 13 chromones, 28 coumarins and four others. Among them, 16 compounds were identified from SR for the first time. In addition, six chromones reference standards, including two isolated compounds of 3'-O-angeloylhamaudol and norcimifugin from the extraction of SR, were used to study the fragmentation pathways of chromones. The developed method was effective for characterising the compounds of SR, and the results of the study enriched the understanding of the chemical connotation.
Subject(s)
Apiaceae/chemistry , Chromatography, High Pressure Liquid/methods , Plant Roots/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Chromones/analysis , Chromones/chemistry , Coumarins/analysis , Coumarins/chemistry , Drugs, Chinese Herbal/chemistry , Medicine, Chinese Traditional , Tandem Mass Spectrometry/methodsABSTRACT
2,3-Dihydro-5-hydroxy-2-methylchromen-4-one (TL1-1) has already been reported to exhibit significant activities such as cytotoxicity, antifungal activity and growth inhibitory activity. In order to simply and efficiently separate TL1-1 from crude extracts of Daldinia eschscholzii on a large-preparative scale, XAD-16 resin was selected from ten types of resin based on its superior adsorption and desorption performance. Adsorption equilibrium data for this resin fitted well with pseudo-first order kinetics and the Freundlich model, which were elucidated from kinetic experiments and adsorption isotherms. Under optimized conditions, the purity of TL1-1 increased from 19.21% (w/w) in the crude extract, to 84.64% (w/w) in the final product, with a recovery yield of 75.06% (w/w) by a one-step treatment. Moreover, in a large-scale separation, the purity and recovery of TL1-1 was 80.33% and 72.02% (w/w), respectively. These results demonstrated that a simple adsorption-desorption strategy, using XAD-16 resin, was efficient, which also highlighted its potential for the future large-scale purification and preparation of TL1-1. In addition, studies showed that the purified TL1-1 exhibited moderate antibacterial activity against Ralstonia solanacearum.
Subject(s)
Anti-Infective Agents/isolation & purification , Chromatography, High Pressure Liquid/methods , Chromones/isolation & purification , Plant Extracts/chemistry , Xylariales/chemistry , Adsorption , Anti-Infective Agents/analysis , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Chromones/analysis , Chromones/metabolism , Chromones/pharmacology , Microbial Sensitivity Tests , Mitosporic Fungi/drug effects , Ralstonia/drug effectsABSTRACT
Yupingfeng (YPF), a famous traditional Chinese medicine, which contains a large array of compounds, has been effectually used in health protection. A two-dimensional liquid chromatography (²D-LC) combined with quadrupole time-of-flight mass spectrometry (QTOF-MS) method was firstly established to separate and identify chemical components in YPF. A total of 33 compounds were identified, including 15 constituents (flavonoids and saponins) in Astragali radix; seven constituents (sesquiterpenoids and polysaccharide) in Atractylodis rhizoma; and 11 constituents (chromone and coumarins) in Saposhnikoviae radix. The corresponding fragmentation pathway of typical substances was investigated. Then, seven active constituents (astragaloside, calycosin, formononetin, cimicifugoside, 4-O-beta-d-glucosyl-5-O-methylvisamminol, sec-O-glucosylhamaudol, and atractylenolide II) derived from three medicinal plants were chosen to further investigate the pharmacokinetic behavior of YPF formula using ultrahigh-performance liquid chromatography with triple quadrupole mass spectrometry system. The method was sensitive, accurate and reliable. We also used the area under the plasma concentration-time curve from zero to infinity (AUC0-∞) as weighting factor to make an integrated pharmacokinetic curve. Results show that the constituents of Saposhnikoviae radix have the best absorption and pharmacokinetic behavior and may play important role in leading to the changes of overall therapeutic effects of YPF. Further study is needed to confirm the association between them.
Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Administration, Oral , Animals , Chromatography, High Pressure Liquid , Chromones/analysis , Coumarins/analysis , Drugs, Chinese Herbal/administration & dosage , Flavonoids/analysis , Male , Medicine, Chinese Traditional , Plant Extracts/chemistry , Polysaccharides/analysis , Rats , Rats, Sprague-Dawley , Saponins/analysis , Tandem Mass SpectrometryABSTRACT
During the development of natural herbal medicines in Japan, Glehniae Radix cum Rhizoma (Hamabofu in Japanese) has been used as a substitute for Saposhnikoviae Radix (Bofu). Bofu and Hamabofu are blended differently in several Kampo formulae. For example, Bofu is included in Jumihaidokuto by a manufacturer, whereas Hamabofu is included instead of Bofu in the same formula by other manufacturers. Although both Bofu and Hamabofu are used for their expected anti-inflammatory effects, differences in their medicinal properties are not well characterized. In addition, there have been very few reports comparing the pharmacological activities of the constituents in Bofu and Hamabofu. In the present study, we investigated the anti-inflammatory effects of the extracts of Bofu and Hamabofu by monitoring levels of the inflammatory mediator nitric oxide (NO) produced in rat hepatocytes. Moreover, the chemical constituents responsible for the activity were investigated. Our results showed that ethyl acetate fractions of Bofu and Hamabofu extracts contain different compounds, although both fractions suppressed NO production in rat hepatocytes. The linear dihydropyranochromones from the Bofu extract (i.e., 3'-O-angeloylhamaudol, ledebouriellol and hamaudol) suppressed NO production, whereas the coumarins from the Hamabofu extract (i.e., umbelliferone and scopoletin) also suppressed NO production. These results suggest that linear dihydropyranochromones and coumarins are responsible for the anti-inflammatory effects of Bofu and Hamabofu. It is plausible that Bofu and Hamabofu are blended differently in several Kampo formulae due to many constituents with as yet unidentified pharmacological activity.
Subject(s)
Anti-Inflammatory Agents/analysis , Apiaceae/chemistry , Chromones/analysis , Coumarins/analysis , Drugs, Chinese Herbal/chemistry , Nitric Oxide/biosynthesis , Animals , Anti-Inflammatory Agents/pharmacology , Cells, Cultured , Chromones/pharmacology , Coumarins/pharmacology , Drugs, Chinese Herbal/pharmacology , Japan , Male , Medicine, Kampo , Plant Roots/chemistry , Rats , Rats, Wistar , Rhizome/chemistryABSTRACT
A new magnetic molecularly imprinted polymers (MMIPs) for quercetagetin was prepared by surface molecular imprinting method using super paramagnetic core-shell nanoparticle as the supporter. Acrylamide as the functional monomer, ethyleneglycol dimethacrylate as the crosslinker and acetonitrile as the porogen were applied in the preparation process. Fourier transform infrared spectrometer (FT-IR), X-ray diffraction (XRD) and Vibrating sample magnetometer (VSM) were applied to characterize the MMIPs, and High performance liquid chromatography (HPLC) was utilized to analyze the target analytes. The selectivity of quercetagetin MMIPs was evaluated according to their recognition to template and its analogues. Excellent binding for quercetagetin was observed in MMIPs adsorption experiment, and the adsorption isotherm models analysis showed that the homogeneous binding sites were distributed on the surface of the MMIPs. The MMIPs were employed as adsorbents in solid phase extraction for the determination of quercetagetin in Calendula officinalis extracts. Furthermore, this method is fast, simple and could fulfill the determination and extraction of quercetagetin from herbal extract.
Subject(s)
Calendula , Chromones/analysis , Molecular Imprinting , Plant Extracts/chemistry , Polymers/chemistry , Acetonitriles/chemistry , Acrylamide/chemistry , Adsorption , Chromones/chemistry , Flavones , Magnetite Nanoparticles/chemistry , Methacrylates/chemistry , Silicon Dioxide/chemistry , Spectroscopy, Fourier Transform Infrared , X-Ray DiffractionABSTRACT
OBJECTIVE: To compare Cirsium japonicum characteristics with C. leo and C. leducei, along with the content of buddleoside and pectolinarin, and lay the foundation for the quality control of C. japonicum. METHOD: Samples were collected and the relevant drugs were bought. The samples were divided into root, stem, leaf and flower, and the content of buddleoside and pectolinarin was determine by the HPLC. Chromatographic column: Waters XBridge C18 (4.6 mm x 250 mm), mobile phase: methanol-water (45: 55), measurement wavelength: 326 nm, flow rate: 0.8 mL x min(-1), column temperature: 30 degrees C. RESULT AND CONDUSION: Standard curve equation of buddleoside: Y = 74 064X-47 748, R2 = 0.991. Standard curve equation of pectolinarin: Y = 1 711 64X - 180 707, R2 = 0.999. The content of buddleoside: C. japonicum leaf was 1.987 3%, C. leo leaf 1.412 2%, C. leducei leaf 0.149 2%. The content of buddleoside was lower in root and stem. Pectolinarin was not detected in the C. japonicum and C. leo. The pectolinarin content was 0.069 0% in C. leducei leaf.
Subject(s)
Chromones/analysis , Cirsium/chemistry , Drugs, Chinese Herbal/analysis , Chromatography, High Pressure Liquid , Chromones/chemistry , Drugs, Chinese Herbal/chemistry , Reproducibility of Results , Solubility , Species SpecificityABSTRACT
In this study, metabolite profiling of Radix Saposhnikoviae from different geographical locations was performed using high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry (HPLC-ESI-TOFMS) and multivariate statistical analysis technique. Principle component analysis (PCA) of the data shows that these samples could be roughly separated into three groups: Guan Fangfeng, Kou Fangfeng and Chuan Fangfeng. The potential chemical markers were discovered through the loading plot of PCA. Based on accurate mass measurements and subsequent fragment ions of TOFMS after in-source collision induced dissociation, as well as matching of empirical molecular formulae with those of published components in the in-house chemical library, 10 potential markers, such as 4'-O-glucosyl-5-O-methylvisamminol, cimifugin, prim-O-glucosylcimifugin and 3'-O-angeloylhammaudol, were tentatively identified and partially verified by the available reference standards. The results of this study indicate that it is an effective and novel approach to identify traditional Chinese medicine (TCM) from different sources, and that performing quantity determination of corresponding marker compounds could optimize the quality control of TCM.
Subject(s)
Apiaceae/chemistry , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Analysis of Variance , Apiaceae/metabolism , Biomarkers/analysis , Biomarkers/chemistry , Chemical Fractionation , Chromones/analysis , Chromones/chemistry , Drugs, Chinese Herbal/standards , Metabolomics , Methanol/chemistry , Monosaccharides/analysis , Monosaccharides/chemistry , Plant Roots/chemistry , Plant Roots/metabolism , Principal Component Analysis , Reproducibility of Results , Xanthenes/analysis , Xanthenes/chemistryABSTRACT
1. Frutinone is an active ingredient extracted from the lipophilic fraction of the Polygala Fruticosa demonstrating various antibacterial and fungal properties. The aim of this study was to characterize its metabolism in an effort to understand metabolism based drug-herb interactions. 2. In vitro metabolic clearance and metabolite identification studies were done using cryopreserved hepatocytes. Reaction phenotyping and inhibition studies were done using human liver microsomes and recombinant cytochrome P450s (CYPs). Frutinone A-CYP1A2 interactions were rationalized using docking simulations. 3. Hepatic clearance was predicted to be low (7.17 mL/min/kg), with reaction phenotyping studies indicating no clearance by the enzymes tested. Frutinone was identified as a potent inhibitor of CYP1A2 with moderate effects on CYP2C19, 2C9, 2D6 and 3A4. CYP1A2 inhibition was reversible and characterised by an IC(50) of 0.56 µM. Inhibition was differential showing mixed (K(i) = 0.48 µM) and competitive (K(i) = 0.31 µM) inhibition with 3-cyano-7-ethoxycoumarin and ethoxyresorufin, respectively. Two binding sites, one for inhibitors and the other for substrates were identified in silico. 4. The potent CYP1A2 inhibition by Frutinone A could be predictive of the potential drug-herb interaction risk in the use of herbal extracts from P. fruticosa. The data suggest future pharmacological research on this chromocoumarin should take metabolic properties into account.
Subject(s)
Anti-Infective Agents/pharmacology , Chromones/pharmacology , Coumarins/pharmacology , Cytochrome P-450 CYP1A2 Inhibitors , Enzyme Inhibitors/pharmacology , Plant Extracts/chemistry , Polygala/chemistry , Anti-Infective Agents/analysis , Anti-Infective Agents/chemistry , Catalytic Domain , Chromones/analysis , Chromones/chemistry , Coumarins/analysis , Coumarins/chemistry , Coumarins/metabolism , Cryopreservation , Cytochrome P-450 CYP1A2/metabolism , Drug Evaluation, Preclinical , Enzyme Inhibitors/analysis , Enzyme Inhibitors/chemistry , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/metabolism , Kinetics , Mass Spectrometry , Metabolic Networks and Pathways/drug effects , Molecular Docking Simulation , Nitriles/metabolism , Oxazines/metabolism , Reproducibility of Results , Substrate Specificity/drug effects , Time FactorsABSTRACT
Arabidopsis thaliana pER8:GUS, a low-cost, highly efficient, and convenient transgenic plant system, was used to assay the estrogen-like activity of 30 traditional Chinese medicines. The MeOH extract of Caesalpinia sappan exhibited significant bioactivity in this assay, and subsequent bioactivity-guided fractionation of the extract led to the isolation of one new compound, (S)-3,7-dihydroxychroman-4-one (1), and 10 known compounds. Both the plant pER8:GUS and in vitro estrogen response element reporter assays were used to evaluate the estrogenic activity of the isolated compounds, and these two systems produced comparable results. Compounds 6, 8, and 11 showed significant estrogenic activity comparable to genistein. These active compounds were determined to be nontoxic new sources of phytoestrogens. In addition, compounds 2 and 3 inhibited ERE transcription induced by 17ß-estradiol. A docking model revealed that compounds 6, 8, and 11 showed high affinity to the estrogen receptor. The pER8:GUS reporter system was demonstrated to be a useful and effective technique in phytoestrogen discovery.
Subject(s)
Caesalpinia/metabolism , Chromones/analysis , Drugs, Chinese Herbal/pharmacology , Estrogens/analysis , Phytoestrogens/analysis , Arabidopsis/genetics , Arabidopsis/metabolism , Chromones/chemistry , Chromones/pharmacology , Drug Screening Assays, Antitumor , Drugs, Chinese Herbal/chemistry , Estradiol/pharmacology , Estrogens/pharmacology , Female , Genistein/pharmacology , Hep G2 Cells , Humans , Models, Molecular , Molecular Structure , Phytoestrogens/chemistry , Phytoestrogens/pharmacology , Plants, Genetically Modified/genetics , Selective Estrogen Receptor Modulators/pharmacology , Taiwan , Wood/chemistryABSTRACT
Methods using high performance liquid chromatography with diode array detection (HPLC-DAD) and tandem mass spectrometry (HPLC-MS/MS) were developed and validated for the simultaneous determination of 5 chromones and 6 coumarins: prim-O-glucosylcimifugin (1), cimifugin (2), nodakenin (3), 4'-O-ß-d-glucosyl-5-O-methylvisamminol (4), sec-O-glucosylhamaudol (5), psoralen (6), bergapten (7), imperatorin (8), phellopterin (9), 3'-O-angeloylhamaudol (10) and anomalin (11), in Radix Saposhnikoviae. The separation conditions for HPLC-DAD were optimized using an Ascentis Express C18 (4.6 mm×100 mm, 2.7 µm particle size) fused-core column. The mobile phase was composed of 10% aqueous acetonitrile (A) and 90% acetonitrile (B) and the elution was performed under a gradient mode at a flow rate of 1.0 mL/min. The detection wavelength was set at 300 nm. The HPLC-DAD method yielded a base line separation of the 11 components in 50% methanol extract of Radix Saposhnikoviae with no interfering peaks detected. The HPLC-DAD method was validated in terms of linearity, accuracy and precision (intra- and inter-day), limit of quantification (LOQ), recovery, and robustness. Specific determination of the 11 components was also accomplished by a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) source. This HPLC-MS/MS method was also validated by determining the linearity, limit of quantification, accuracy, and precision. Quantification of the 11 components in 51 commercial Radix Saposhnikoviae samples was successfully performed using the developed HPLC-DAD method. The identity, batch-to-batch consistency, and authenticity of Radix Saposhnikoviae were successfully monitored by the proposed HPLC-DAD and HPLC-MS/MS methods.
Subject(s)
Apiaceae/chemistry , Chromatography, High Pressure Liquid/methods , Chromones/analysis , Coumarins/analysis , Plant Extracts/chemistry , Linear Models , Reproducibility of Results , Sensitivity and Specificity , Tandem Mass SpectrometryABSTRACT
Four chromones, prim-O-glucosylcimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, cimifugin and sec-O-glucosylhamaudol, were isolated and purified from Saposhnikovia divaricata for the first time by high-performance counter-current chromatography (HPCCC) using a system consisting of ethyl acetate/n-butanol/ethanol/water (1:1:0.1:2, v/v/v/v). The separation parameters were first performed on the analytical HPCCC and the optimized conditions were then scaled up to preparative HPCCC. A total of 72.1 mg of prim-O-glucosylcimifugin, 27 mg of 4'-O-ß-D-glucosyl-5-O-methylvisamminol, 14.1 mg of cimifugin and 1.1 mg of sec-O-glucosylhamaudol were purified from 960 mg of the n-butanol extract of S. divaricata, each at over 90% purity as determined by high-performance liquid chromatography (HPLC). The structures of four compounds were identified by their retention time, the liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) in the positive ion mode, and confirmed by NMR. The characteristic LC-ESI-MS fragmentation patterns of the four compounds were discussed, and found to be a very specific and useful tool for the structural identification of chromones from S. divaricata.
Subject(s)
Apiaceae/chemistry , Chromatography, High Pressure Liquid/methods , Chromones/isolation & purification , Plant Extracts/isolation & purification , Chromones/analysis , Plant Extracts/analysisABSTRACT
INTRODUCTION: The popular use of black cohosh products (Actaea racemosa L., syn. Cimicifuga racemosa L.) is growing as the demand for alternatives to estrogen therapy has increased. Critical to safe use is the assurance of unadulterated, high-quality products. Questions have been raised about the safety of black cohosh due to cases of liver toxicity in patients who reported taking it; subsequent evaluation found some products to be adulterated with other related herbal species. Correct plant species identification is a key first step for good manufacturing practices of safe black cohosh products. OBJECTIVES: To develop analytical methods which distinguish black cohosh from other species (American and Asian) of Actaea increasingly found as adulterants in commercially available black cohosh products. MATERIAL AND METHODS: Fifteen species of Actaea were collected from North America and Asia, and the phytochemical fingerprints of these samples were established using HPLC-PDA and LC-MS techniques. RESULTS: The HPLC and LC-MS fingerprints for polyphenols and triterpene glycosides revealed distinct patterns that make black cohosh clearly distinguishable from most other species of Actaea. Two marker compounds, cimifugin and cimiracemoside F, were found to be important to distinguish black cohosh from most Asian species of Actaea. Formononetin was not found from either Asian or American species of Actaea. CONCLUSIONS: Phytochemical fingerprinting is a practical, reliable method for authenticating black cohosh and distinguishing it from other species of Actaea increasingly found as adulterants in commercially available black cohosh products. This should facilitate the continued development of high-quality, unadulterated black cohosh products.