No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Simultaneous Determination of Nine Major Flavonoids in Sophora flavescens by RP-LC
Abstract
A liquid chromatographic method was applied to determine trifolirhizin, kushenol K, kushenol L, kushenol N, kushenol X, kurarinone,
norkurarinone, isokurarinone and kushenol A in the roots of Sophora flavescens, namely Kushen in China. The samples were separated on a YMC-C18 column (250×4.6mm, 5μm) with a gradient of methanol and 0.3% aqueous acetic acid (v/v) at a flow rate of 0.8mLmin−1 and detected at 295nm. The complete separation was achieved within 45min for the nine major flavonoids. All calibration
curves expressed good linearity (r
2>0.999) within the test range. The recovery of this method was 92.3–106.9%. The assay was successfully applied to the quantification
of nine flavonoids in 26 samples of Kushen. The results indicated that this developed LC assay could be readily utilized as
a quality control method for the Chinese herb medicine Kushen.
... Kurarinone is one of the major constituents of the traditional Chinese medicine Sophorae Flavescentis Radix, or Kushen in Chinese (the root of Sophora flavescens), which is often used to treat diarrhea, bacterial and fungal infections, eczema, and inflammation-related diseases (20). Kurarinone shares a flavanone core with a characteristic lavandulyl moiety at C-8 (21,22) and possesses several pharmacological effects, such as anti-inflammatory and ...
... As shown in SI Appendix, Fig. S5, six and four peaks were identified in the LC-MS/MS plot of plasma and brain, namely kushenols F, N, and S, flemiphilippinin D, sophoraflavanone G, and kurarinone, respectively. Kurarinone (Fig. 1A), the major constituent of SFE (22), was used to assay for its anti-PD effect in MPTP-treated mice. To assess the effect of kurarinone on MPTP-induced gait disorders, some behavioral parameters were also measured on the CatWalk Automated Gait Analysis System, including duration, maximum variation, average speed, and cadence. ...
Significance
To date, no cure or preventative treatment for Parkinson’s disease (PD) has yet been developed. Here, we show that kurarinone, a natural flavonoid, alleviated parkinsonism-like symptoms induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice. Using a proteomics approach, we identified the soluble epoxide hydrolase (sEH) as a possible target of kurarinone’s reduction of neuroinflammation. This was supported using complementary biochemical approaches, which demonstrated that kurarinone is a high nanomolar uncompetitive inhibitor of sEH. Our findings suggest that natural products could attenuate the development of PD through inhibition of sEH.
... The main bioactive component of Sophora flavescens is oxymatrine, which possesses important biological activities, such as anticancer and inhibition of hepatitis B virus replication567. Oxymatrine can be extracted from this plant by several methods, such as maceration extraction, the decocting, refluxing or leakage methods and supercritical fluid extraction89101112. Although methanol-water and chloroform-ammonia solutions were also used as extracting solvents for extraction of oxymatrine from Sophora flavescens in the literature [8,13], water and ethanol-water solutions were more widely used because of the toxicity of chloroform and methanol, which are harmful to human beings and environment. ...
... Although methanol-water and chloroform-ammonia solutions were also used as extracting solvents for extraction of oxymatrine from Sophora flavescens in the literature [8,13], water and ethanol-water solutions were more widely used because of the toxicity of chloroform and methanol, which are harmful to human beings and environment. The extraction temperature range used in the literature was from room temperature (about 25 °C) to 100 °C89101112. However, the extraction rates of oxymatrine were usually low using conventional methods, such as the decocting method (52.3%) and refluxing method (53.4%) [8]. ...
In this paper, microwave-assisted extraction (MAE) of oxymatrine from Sophora flavescens were studied by HPLC-photodiode array detection. Effects of several experimental parameters, such as concentration of extraction solvent, ratio of liquid to material, microwave power, extraction temperature, and extraction time on the extraction efficiencies of oxymatrine were evaluated. The optimal extraction conditions were 60% ethanol, a 20:1 (v/v) ratio of liquid to material and extraction for 10 min at 50 °C under 500 W microwave irradiation. Under the optimum conditions, the yield of oxymatrine was 14.37 mg/g. The crude extract obtained could be used as either a component of some complex traditional medicines or for further isolation and purification of bioactive compounds. The results, which indicated that MAE is a very useful tool for the extraction of important phytochemicals from plant materials, should prove helpful for the full utilization of Sophora flavescens.
... Mostly, single marker compound is analyzed to evaluate the quality of TCMs [3], which is simple but cannot totally demonstrate the quality of herbal prescriptions. Then, multiple components analysis (MCA) method has been developed, which can simultaneously evaluate many active compounds from different herbs and has been widely used for the quality control of TCMs [4][5][6]. ...
In this study, a simple, reliable and accurate method for the simultaneous separation and determination of naringin, hesperidin, neohesperidin, baicalin, wogonoside, baicalein, wogonin, emodin and chrysophanol in 'Da-Chai-Hu-Tang' was developed by reverse-phase high-performance liquid chromatography (RP-HPLC). The chromatographic separation was performed on an Agilent ZORBAX C18 column (250 mm × 4.6 mm i.d., 5.0 μm), and the mobile phase composed of methanol and water containing 1% (v/v) acetic acid was used to elute the targets in a gradient elution mode. The flow rate and detection wavelength were set at 0.8 ml/min and 280 nm, respectively. All calibration curves of the nine components expressed good linearities (r2 ≥ 0.9992) within the tested ranges. The RSD values demonstrated the intra-and inter-day precisions were less than 2.89%, and the recoveries of the investigated compounds were between 96.22% and 105.28%. The proposed method is simple, precise, specific, sensitive, and successfully applied to determine the nine marker compounds in 'Da-Chai-Hu-Tang' for quality control.
Monoamine oxidase (MAO) catalyzes the oxidation of monoamines and its two isoforms, MAO-A and MAO-B, break down neurotransmitter amines. Of the compounds isolated from the roots of Sophora flavescens, (-)-maackiain (4), a pterocarpan, was found to potently and selectively inhibit human MAO-B, with an IC50 of 0.68μM, and to have a selectivity index of 126.2 for MAO-B. As compared with other herbal natural products, the IC50 value of 4 for MAO-B is one of the lowest reported to date. Genistein (1) highly, effectively and non-selectively inhibited MAO-A and MAO-B with IC50 values of 3.9μM and 4.1μM, respectively. (-)-4-Hydroxy-3-methoxy-8,9-methylenedioxypterocarpan (2) effectively and non-selectively inhibited MAO-A and MAO-B with IC50 values of 20.3μM and 10.3μM, respectively. In addition, compound 4 reversibly and competitively inhibited MAO-B with a Ki value of 0.054μM. Molecular docking simulation revealed that the binding affinity of 4 for MAO-B (-26.6kcal/mol) was greater than its affinity for MAO-A (-8.3kcal/mol), which was in-line with our inhibitory activity findings. Furthermore, Cys172 of MAO-B was found to be a key residue for hydrogen bonding with compound 4. The findings of this study suggest compound 4 be viewed as a new potent, selective, and reversible MAO-B inhibitor, and that compounds 1 and 2 be considered useful lead compounds for the developments of nonselective and reversible MAO inhibitors for the treatment of disorders like Parkinson's disease, Alzheimer disease, and depression.
In this study, a simple, sensitive, and robust analytical method based on ultra-performance liquid chromatography (UPLC) has been developed for the determination of trifolirhizin in rat plasma using pirfenidone as internal standard (IS). After sample preparation by a simple liquid-liquid extraction, chromatography was performed on an Acquity UPLC BEH C18 column (2.1mm×50mm, 1.7μm particle size) and ultraviolet detection set at a wavelength of 366nm. The method was linear over the concentration range 25-1000ng/mL with a lower limit of quantification (LLOQ) of 25ng/mL. Inter- and intra-day precision (RSD%) were all within 10.2% and the accuracy (RE%) was equal or lower than 9.3%. The recovery was in the range of 78.5-86.4% for trifolirhizin and 87.4% for IS. Stability studies showed that trifolirhizin was stable under a variety of storage conditions. The method was successfully applied to a pharmacokinetic study involving oral administration of trifolirhizin to rats.
Copyright © 2015 Elsevier B.V. All rights reserved.
Zhixue capsule is a prescription for hemorrhoid commonly used in traditional Chinese medicine. This drug was recalled by the State Food and Drug Administration in 2008 because of severe adverse hepatic reactions. Zhixue capsule is composed of ethanol extracts of Cortex Dictamni (ECD) and Sophora flavescens (ESF). In our preliminary study, we observed the hepatotoxic effects of ESF on rat primary hepatocytes. However, ECD did not exhibit hepatotoxicity at the same concentration range. In this study, ESF was evaluated for its potential hepatotoxic effects on rats. Bioassay-guided isolation was used to identify the material basis for hepatotoxicity. Treatment with 1.25 g/kg and 2.5 g/kg ESF significantly elevated the alanine aminotransferase and aspartate aminotransferase levels in the serum. The changes in the levels of transaminases were supported by the remarkable fatty degeneration of liver histopathology. Further investigations using bioassay-guided isolation and analysis indicated that prenylated flavanones accounted for the positive hepatotoxic results. Two isolated compounds were identified, kurarinone and sophoraflavanone G, using nuclear magnetic resonance and mass spectrometry techniques. These compounds have potent toxic effects on primary rat hepatocytes (with IC50 values of 29.9 μM and 16.5 μM) and human HL-7702 liver cells (with IC50 values of 48.2 μM and 40.3 μM), respectively. Consequently, the hepatotoxic constituents of S. flavescens were determined to be prenylated flavanones, kurarinone, and sophoraflavanone G.
A preparative high-speed counter-current chromatography (HSCCC) method for isolation and purification of four prenylflavanones from the crud extract after microbial biotransformation of kurarinone was developed successfully using a stepwise elution with two-phase solvent system composed n-hexane–ethyl acetate–methanol–water at the volume ratios of 1:1:0.7:1 (v/v) and 1:1:1.2:1 (v/v). A total of 7mg of 4a,5a-dihydroxy norkurarinone (1), 11mg of 7-methoxyl-4a,5a-dihydroxy norkurarinone (2), 14mg of 6a-hydroxykurarinone (3) and 10mg of norkurarinone (4) were obtained in one-step separation from 520mg of the crude biotransformation sample with purities of 94.0%, 97.8%, 99.5% and 98.7%, respectively. Among them, compounds 1–3 are novel compounds, and their chemical structures were identified on the basis of the extensive spectral methods such as UV, IR, HR-MS and NMR.
A simple, reliable and accurate high performance liquid chromatography(HPLC) coupled with a UV detector at varied wavelength was developed for the quantitation of two types of anti-carcinogenic compounds, namely, Kushen alkaloids(KS-As) and Kushen flavonoids(KS-Fs), in Kushen. Their remarkable difference in physicochemical properties was circumvented by integrated optimization of column and mobile phase. The separation was achieved on an alkaline-resisting C-18 column via gradient elution with acetonitrile(ACN) and 0.025%(volume ratio) diethylamine(DEA) in water as mobile phase. The flow rate was 0.7 mL/min, and the column temperature was maintained at 35 C. UV detection wavelength was set at 225 nm to monitor KS-As, and then changed to 335 nm for KS-Fs at 26th min, which was kept for 30 min before returning to the original wavelength. All the calibration curves show good linearity(r(2)> 0.999) within test ranges. The intra- and inter-day precision values of these components were less than 3.82%, and the average recovery rates obtained were in a range of 97.24%-102.64% for all the samples with RSD below 3.94%. This assay was successfully applied to determining the major active components in commercial herbal samples of Kushen. Moreover, the possible relationship between sample and the geographical region was visualized based on the contents of the 6 compounds in the light of principal component analysis(PCA). The results suggest that the proposed method facilitates the quality control of Kushen.
In this paper, microbial transformation of kurarinone (1) by Cunninghamella echinulata AS 3.3400 was investigated and four transformed products were isolated and identified as 6″-hydroxykurarinone (2), 4″,5″,8″-trihydroxynorkurarinone (3), norkurarinone (4), and kurarinone-7-O-β-glucoside (5), respectively. Among them, 3 and 5 are new compounds, and the rare glycosylation in microbial transformation was observed. In addition, the cytotoxicities of transformed products (2-5) were also investigated.
In this paper, microbial transformation of norkurarinone (1) by Cunninghamella blakesleana AS 3.970 was investigated and seven transformed products were isolated and characterized as kurarinone (2), 4″,5″-dihydroxykurarinone (3), 6″-hydroxyl-2'-methoxyl-norkurarinone 7-O-β-d-glucoside (4), 6″-hydroxyl-norkurarinone 4'-O-β-d-glucoside (5), 4″,5″-dihydroxynorkurarinone (6), 7-methoxyl-norkurarinone (7), and 7-methoxyl-4″,5″-dihydroxynorkurarinone (8), respectively. Among them, 3-5 are new compounds, and the glycosylation reaction in microbial transformation process was reported rarely. In addition, the cytotoxicities of transformed products (1-8) were also investigated.
Two novel lavandulyl flavonoids, (2S)-7-methoxyl-4",5"-dihydroxynorkurarinone (1) and (2S)-6"-hydroxynorkurarinone-7-O-beta-D-galactoside (2), were isolated from the rhizome of Sophora flavescens. Their structures were elucidated by spectral methods, including 2D NMR spectroscopy. Both compounds showed cytotoxic activity against Hela cells, with 2 being more active than 1.
The biological activity of ten prenylflavanones purified from Sophora tomentosa L., and Sophora moorcroftiana Benth. ex Baker (Leguminosae) was investigated. The flavanones with prenyl-, lavandulyl- or geranyl groups on A ring, and two bioactive flavonostilbenes on ring B and stilbene (resveratrol) showed tumor-specific cytotoxic activity, antimicrobial activity, and anti-HIV activity, radical generation, and O2- scavenging activity. There was a positive relationship between radical generation and O2- scavenging activity in these prenylflavanones. These data suggest the medicinal significance of prenylflavanones.
The methanol extract of Sophora flavescens showed a potent glycosidase inhibitory activity. Active components were identified as well-known flavonoid antioxidants: kushenol A (1), (-)-kurarinone (2), sophoraflavanone G (3), 2'-methoxykurarinone (4), kurarinol (5), 8-prenylkaempferol (6), isoxanthohumol (7), kuraridin (8) and maackian (9). All flavonoids were effective inhibitors of alpha-glucosidase and beta-amylase. Interestingly, lavandulylated flavanones 1-5 had strong alpha-glucosidase inhibitory activities, with IC(50) values of 45 microM, 68 microM, 37 microM, 155 microM and 179 microM, respectively. Kushenol A (1) which does not bear a 4'-hydroxy group showed selective alpha-glucosidase inhibitory activity. Lavandulylated chalcone, kuraridine (8), exhibited IC(50) value of 57 microM against beta-glucosidase, which is the first report of a chalcone displaying glycosidase inhibition. Results showed that 8-lavandulyl group in B-ring was a key factor of the glycosidase inhibitory activities. The inhibition pattern was noncompetitive for alpha-glucosidase, whereas mixed inhibition was observed for beta-amylase.
Four new flavanonols, kushenol J (I), kushenol K (II), kushenol L (III) and kushenol M (IV), were isolated from the dry roots of Sophora flavescens AIT. (Leguminosae). Their structures were elucidated on the basis of spectral data especially ¹H nuclear magnetic resonance and ¹³C nuclear magnetic resonance. It is the first case that the isolation of a flavonoid glycoside (I) and a cis flavanonol (II) derivatives from Sophora flavescens AIT.
A new benzoquinone, kushequinone A (I), a new flavanonol, kushenol N (II), and a new isoflavone glycoside, kushenol O (III), were isolated from the dry roots of Sophora flavescens AIT. (Leguminosae). Their structures were elucidated on the basis of spectral data especially ¹H nuclear magnetic resonance and ¹³C nuclear magnetic resonance. A benzoquinone derivative was isolated for the first time from Sophora flavescens AIT.
Two new flavanones, I and VII, and daidzein were isolated from Guang-Dou-Gen (the root of Sophora subprostrata CHUN et T.CHEN). Fom the spectral data and comparison with sophoranochromene and its derivative, the structure of I was established as 2-[{3'-hydroxy-2', 2'-dimethyl-8'-(3-methyl-2-butenyl)} chroman-6'-yl]-7-hydroxy-8-(3-methyl-2-butenyl) chroman-4-one. The structure of VII was formulated from the spectral data and biogenetic speculation as 2-[{2'-(1-hydroxy-1-methylethyl)-7'-(3-methyl-2-butenyl)-2'3'-dihydrobenzofuran}-5'-yl]-7-hydroxy-8-(3-mehtyl-2-butenyl) chroman-4-one.
Two flavanones, kushenol A (1) and kushenol B (2), one flavonol, kushenol C (3), one chalcone, kushenol D (4), and one pterocarpan, kushenin (5), were isolated from the dry roots of Sophora flavescens AITON (Leguminosae). Their structures were elucidated on the basis of elemental analyses and spectral data.
Two new lavandulylated flavanones, (2S)-2'-methoxykurarinone (1) and (-)-kurarinone (2), were isolated from the root of Sophora flavescens, together with two known lavandulyl flavanones, sophoraflavanone G (3) and leachianone A (4), and two known isoflavonoids, formononetin and l-maakiain. The structures of 1 and 2 were determined on the basis of optical rotation and spectral evidence and by comparison with known compounds. Compounds 1-4 exhibited cytotoxic activity against human myeloid leukemia HL-60 cells.
Three new flavonoids, which are isoprenylated by fused 2,2-dimethyl-3,4-dihydro-2H-pyran moieties, were isolated from the roots of Sophora flavescens and named flavenochromanes A–C (1–3). Their structures were elucidated by spectroscopic methods, including 2D-NMR techniques. Flavenochromane C (3) showed strong cytotoxic activity against A549 (lung carcinoma), 1A9 (ovarian carcinoma), KB (epidermoid carcinoma of the nasopharynx), and KB-Vin (drug-resistant variant KB) cell lines with IC50 values ≤1.7 μM, and significant activity against the MCF-7 (breast adenocarcinoma) cell line with an IC50 value of 3.6 μM. Flavenochromane B (2) displayed slightly lower inhibitory effects (IC50 3.2–6.9 μM) as compared with 3.
A rapid and accurate HPLC method has been developed for the simultaneous determination of nine major flavonoids, namely 3-hydroxymelanettin, melanettin, stevenin, butein, isoliquiritigenin, dalbergin, 2,4-dihydroxy-5-methoxybenzophenone, pinocembrin, 4-methoxydalbergione in Dalbergia odorifera. The samples were separated on an Aglient Zorbax SB-C18 column (250 4.6 mm, 5 m) with a gradient of acetonitrile and 1% aqueous acetic acid (/) at a flow rate of 1.0 mL min–1 and detected at 350 nm. The complete separation was obtained within 30 min for the nine target flavonoids. All calibration curves showed good linearity (r2 > 0.999) within test ranges. The repeatability was evaluated by intra- and inter-day assays and RSD values were less than 4.0%. The recoveries were between 92.0% and 104.5%. The developed method was successfully applied to the analysis of 32 commercial samples of D. odorifera.
All fifteen flavonoids (1 approximately 15) have been isolated from the roots ofSophora flavescens (Leguminosae) as active principles of the cytotoxic property toward human tumor cell lines such as A549, SK-OV-3, SK-Mel-2, XF498 and HCT15,in vitro. By means of spectral analyses, particularly by the aid of various two dimensional NMR experiments, all(1)H-NMR and(13)C-NMR signals of1 approximately 15 were completely assigned, and thus the structures of1 approximately 15 were established unambiguously.
A reversed phase high performance liquid chromatography method was established for the simultaneous determination of eight
major constituents, namely gallic acid, paeoniflorin sulfonate, catechin, albiflorin, paeoniflorin, benzoic acid, pentagalloylglucose
and benzoylpaeoniflorin in red and white peony roots, the two commonly used traditional Chinese medicinal herbs. The optimal
conditions of separation and detection were achieved on a C18 analytical column with a gradient mobile phase consisting of acetonitrile and 0.015% phosphoric acid at the flow rate of
1.0 mL min−1 and detection wavelength set at 230 nm. All calibration curves showed good linear regression (r>0.9995) within test ranges.
This method provided good reproducibility with overall intra-and inter-day precision of less than 5% and 4% and good accuracy
with recovery of more than 93%, respectively. The method was successfully applied to determine 71 samples of red and white
peony roots collected from different areas. The results indicated that the contents of eight compounds varied significantly
among the samples determined, which mainly resulted from processing procedure and habitat variation. The roots of Paeonia veitchii Lynch. contained a much higher amount of gallic acid and pentagalloylglucose than that of Paeonia lactiflora Pall., which could be used to distinguish the two similar species.
The stem of Kadsura heteroclita is a traditional Chinese medicine with a variety of biological activities. For efficient monitoring of the quality of the
herb, a simple, rapid, and accurate HPLC method has been developed for simultaneous determination of five cyclolanostane triterpenoid
compounds (schisanlactone E, nigranoic acid, schisandronic acid, and heteroclitalactones A and D) and one tetrahydrofuran
lignan (d-epigalbacin) in the stems of K. heteroclita. These six compounds were separated on a C18 column by elution with a mobile-phase gradient prepared from acetonitrile and 0.05% (v/v) aqueous phosphoric acid. The flow rate was 1.0mLmin−1 and the detection wavelength was 210nm. The recovery of the method was in the range 95.03–102.19% and a good linear relationship
(r
2>0.9999) was obtained between response and compound concentration over a relatively wide range. The method was successfully
applied to analysis of the herb collected at different stages of growth.
A new capillary electrophoresis (CE) method for the determination of quinolizidine alkaloids in Sophora medicinal plants was developed. A total of seven alkaloid components (cytisine, sophocarpine, matrine, lehmannine, sophoranol, oxymatrine and oxysophocarpine) were separated within 15 min. The running buffer was a 50 mM phosphate buffer containing 1%HP-β-CD and 3.3% isopropanol. The linear calibration ranges were 5.50–88.0 μg ml−1 for cytisine and lehmannine, 5.00–88.0 μg ml−1 for sophocarpine and sophoranol, 5.60–89.6 μg ml−1 for matrine and oxysophocarpine, and 24.0–384 μg ml−1 for oxymatrine. The recoveries of the seven alkaloids were 96.0–102.9% with relative standard deviations from 1.50 to 3.00% (n = 5). The method was successfully applied to different Sophora medicinal plants including Sophora flavescens, Sophora tonkinensis and Sophora alopecuroides.
From the Chinese crude drug, "Kushen"(Japanese name "Kujin") (the root of Sophora flavescens AIT.) five new flavonoids, kuraridinol (I), kurarinol (II), neokurarinol (III), nor-kurarinol (IV), isokurarinone (V) (each named by us) and formononetin were isolated, whose structures have been established by spectral and chemical data.
A methanol extract of hops of Humulus lupulus (L.) showed inhibitory activity against rat liver diacylglycerol acyltransferase (DGAT). From DGAT inhibitory activity-guided fractionation, two chalcones were isolated. One was identified as xanthohumol and the other was found to be a new product designated xanthohumol B. The structure of xanthohumol B was shown to be 6-[3,4-dihydro-3,5-dihydroxy-7-methoxy-2,2-dimethyl-2H-benzo[b]pyrano ]- 3-(4-hydroxyphynyl)-2-propen-l-one by spectroscopic studies including various NMR measurements. Xanthohumol and xanthohumol B inhibited DGAT activity with IC50 values of 50.3 and 194 microM in rat liver microsomes, respectively. They showed preferential inhibition of triacylglycerol formation in intact Raji cells, indicating that they inhibit DGAT activity preferentially in living cells.
A new prenylated flavonol, sophoflavescenol (1), together with five known flavonoids, kurarinol, kushenol K, kushenol H, trifolirhizin, and kuraidin, were isolated from the roots of Sophora flavescens. The structure of 1 was determined by spectroscopic analysis. Among the five known flavonoids, kurarinol, kushenol K, and kushenol H showed weak antiviral activity against Herpes simplex virus types I and II.
Sixteen flavanones, three flavanonols, and four pterocarpans were isolated from the MeOH extract of the roots of Sophora flavescens. Twelve of these were new compounds, including eight prenylflavanones (1-8), one prenylflavanonol (9) and three novel pterocarpane derivatives (10-12). Their structures were elucidated using NMR and mass spectral methods. Some of these compounds have irregular C10 prenyl moieties at C-8 of the flavanone skeleton. These compounds exhibited significant antibacterial activities against the Gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, S. epidermidis, and Propionibacterium acnes. They also exhibited antiandrogen activities.
During the search for naturally occurring cyclic guanosine monophosphate (cGMP)-specific phosphodiesterase type 5 (PDE5) inhibitors, it was found that the extracts from Sophora flavescens exhibit potent inhibitory activity against cGMP PDE5 prepared from rat diaphragm. Therefore, the inhibitory activities of five flavonoids, kushenol H (1), kushenol K (2), kurarinol (3), sophoflavescenol (4) and kuraridine (5), isolated from S. flavescens were measured against cGMP PDE5 to identify potent cGMP PDE5 inhibitory constituents. Among tested compounds, sophoflavescenol (4), a C-8 prenylated flavonol, showed the most potent inhibitory activity (IC(50)=0.013 microM) against cGMP PDE5 with 31.5- and 196.2-fold selectivity over PDE3 and PDE4, respectively. Kinetic analysis revealed that sophoflavescenol was a mixed inhibitor of PDE5 with a K(i) value of 0.005 microM.
A simple and rapid capillary zone electrophoresis method was developed for the separation of the main alkaloids from Sophora flavescens Ait. with the optimum buffer solution containing 110 mM NaH(2)PO(4) and 15% 2-propanol (pH 3.0). The field-amplified sample stacking (FASS) technique was applied to the on-line concentration of the alkaloids. The data presented in this work demonstrate that the use of a short water plug at the column inlet is essential for improving the reproducibility of FASS with electro-injection, and that the water plug injection time affected the sensitivity significantly. The sample concentration was further increased by about 2-3-fold by the introduction of a relatively longer water plug. With this stacking measure, the concentration sensitivity was about 3-4 orders of magnitude higher than in hydrodynamic injection.
To find new antipruritic herbal medicines for pruritus, we screened the methanol extracts of seven herbal medicines which have been used to treat dermatologic diseases, testing them on mouse models of acute and chronic itch. When administrated perorally (p.o.) at a dose of 200 mg/kg, methanol extracts of Sophora flavescens and Cnidium monnieri, but not the others, significantly inhibited a serotonin (5-HT)-induced itch-related response (scratching) and the spontaneous scratching of NC mice, a mouse model of atopic dermatitis. The inhibitory effect of Sophora flavescens was stronger than that of Cnidium monnieri. The methanol extract from Sophora flavescens (50-200 mg/kg) inhibited 5-HT-induced scratching in a dose-dependent manner, without any effects on the locomotor activity. These results suggest that Sophora flavescens and its constituents widely affect acute and chronic pruritus, and are possible as new antipruritic agents.
Prenylated flavonoids containing the resorcinol moiety were isolated as tyrosinase inhibitors from the roots of Sophora flavescens by activity-guided fractionation. Among the 12 compounds isolated, kuraridin, kurarinone and norkurarinol showed stronger inhibitory potencies (IC 50 = 1.1, 1.3 and 2.1 microM, respectively) than that of kojic acid (IC 50 = 11.3 microM), a well known tyrosinase inhibitor. Substitution of a lavandulyl or hydroxylavandulyl group at the C-8 position and a methoxy or hydroxy group at the C-5 position are essential for the inhibitory effect.
Antimicrobial activity of the 18 prenylated flavonoids, which were purified from five different medicinal plants, was evaluated by determination of MIC using the broth microdilution methods against four bacterial and two fungal microorganisms (Candida albicans, Saccaromyces cerevisiae, Escherichia coli, Salmonella typhimurium, Staphylococcus epidermis and S. aureus). Papyriflavonol A, kuraridin, sophoraflavanone D and sophoraisoflavanone A exhibited a good antifungal activity with strong antibacterial activity. Kuwanon C, mulberrofuran G, albanol B, kenusanone A and sophoraflavanone G showed strong antibacterial activity with 5-30 microg/ml of MICs. Morusin, sanggenon B and D, kazinol B, kurarinone, kenusanone C and isosophoranone were effective to only gram positive bacteria, and broussochalcone A was effective to C. albicans. IC50 values of papyriflavonol A, kuraridin, sophoraflavanone D, sophoraisoflavanone A and broussochalcone A in HepG2 cells were 20.9, 37.8, 39.1, 22.1, and 22.0 microg/ml, respectively. These results support the use of prenylated flavonoids in Asian traditional medicine to treat microbial infection and indicate a high potential for prenylated flavonoids as antimicrobial agents as well as anti-inflammatory agents.
A reversed-phase liquid chromatographic method was developed for the quantitative determination of six triterpenoids, namely ganoderic acids C2, B, AM1, K, H and D in Ganoderma lucidum and its related species. Samples were extracted with chloroform in ultrasonic bath. The optimal conditions of separation and detection were achieved on an Agilent Zorbax SB-C18 column (250 mmx4.6 mm, 5 microm), with a linear gradient of acetonitrile and 0.03% aqueous phosphoric acid (v/v), at a flow rate of 1.0 ml/min, detected at 252 nm. All calibration curves showed good linearity (r2>0.999) within test ranges. The relative deviation of this method was less than 2% for intra- and inter-day assays, and the percentage recovery of the method was 93-103%, with relative standard deviation (R.S.D.) less than 5%. The current assay method was applied to quantitative determination of constituents of triterpenoids in 36 different samples of G. lucidum and its related species. The results indicated that the developed method could be readily utilized as a quality control method for G. lucidum and related species.
The objectives of this study were to investigate the radical-scavenging activity and protective potential of Sophora flavescens from oxidative damage by the radical generator 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) in renal epithelial LLC-PK(1) cells and to identify the active components using the bioassay-linked fractionation method. The MeOH extract and fractions of CH(2)Cl(2), BuOH, and H(2)O from S. flavescens showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging effects in a dose-dependent manner (p<0.01),whereas only the BuOH and CH(2)Cl(2) fractions showed protective effects against LLC-PK(1) cellular damage induced by AAPH in a dose-dependent manner (p<0.01). In particular, the BuOH fraction had the most effective (p<0.05) antioxidative capacity. Employing a bioassay-linked HPLC/MS method, the active constituents from the BuOH fraction of S. flavescens were isolated and characterized as sophoraflavanone G and kurarinone with potent antioxidant effects against the DPPH radical, with IC(50) values of 5.26 and 7.73 microg/ml, respectively. Moreover, the compounds dose dependently recovered cell viability decreased by AAPH treatment (p<0.01), suggesting their protective roles against cellular oxidative damage. The results of this study suggest that S. flavescens has excellent antioxidative and kidney-protective potential and that flavonoids from S. flavescens, i.e., sophoraflavanone G and kurarinone, are the active constituents.
High-performance liquid chromatographic (HPLC) fingerprints were developed for identification of both lipophilic and hydrophilic components of the roots of Salvia miltiorrhiza and four related preparations. These samples were separated with an Agilent Zorbax Extend C(18) reserved-phase column (5 microm, 250 mm x 4.6 mm) by linear gradient elution using water-phosphoric acid (100:0.026, v/v) and acetonitrile as mobile phase. The flow rate was 0.8 ml/min and the detector wavelength was set at 280 nm. Mean chromatograms and correlation coefficients of samples were calculated by the software "Similarity Evaluation System for Chromatographic Fingerprint of TCM". The correlation coefficients of Danshen and Fufang Danshen tablets (FDT) samples were in the range of 0.352-0.993 and 0.768-0.987, respectively. The correlation coefficients of Compound Danshen dripping pills (CDDP), Danshen injection (DSI) and Xiangdan injection (XDI) samples were higher than 0.928, 0.850 and 0.960, respectively. It was the first time to identify 34 peaks by comparing with standard compounds and using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS(n)) technique. All results indicated that the developed fingerprint assay could be readily utilized as a quality control method for S. miltiorrhiza and its related preparations.
A high performance liquid chromatographic method was developed to simultaneously determine the 12 major constituents of Forsythia suspensa, namely R-suspensaside, S-suspensaside, S-suspensaside methyl ether, (+)-pinoresinol-beta-d-glucoside, forsythiaside, (+)-epipinoresinol-4'-O-glucoside, suspensaside A, rutin, phillyrin, (+)-pinoresinol, (+)-epipinoresinol and phillygenin. The HPLC assay was performed on a Zorbax XDB C(18) column with gradient elution of methanol and 0.3% aqueous acetic acid within 55 min. The detection wavelength was 280 nm. All the compounds showed good linearity (r(2)>0.9998). The method was reproducible with intra- and inter-day variation less than 3%. The recovery of the assay was in the range of 91.2-104.9%. The method was successfully applied to the quantification of 12 constituents in 33 F. suspensa samples. The results indicated that the developed assay could be considered as a suitable quality control method for F. suspensa.
A new dihydroflavonol named kosamol A (1) has been isolated from the roots of Sophora flavescens (Leguminosae) along with twelve related flavonoids. The structure of 1 was determined to be (2R,3R)-5,7,2'4'-tetrahydroxy-6-(3-hydroxy-3-methylbutyl)-8-lavandulylflavanonol on the basis of spectral analyses.
The methanol extract of Sophora flavescens, which is used in traditional Chinese medicine (sophorae radix), showed potent Na(+)-glucose cotransporter (SGLT) inhibitory activity. Our search for active components identified many well-known flavonoid antioxidants: kurarinone, sophoraflavanone G, kushenol K, and kushenol N.
doi:10.1016/j.jchromb
- Ah Liu
- Yh Lin
- M Yang
- Hz Guo
- Sh Guan
- Sun
doi:10.1016/j.phymed
- Hy Sohn
- Kh Son
- Cs Kwon
- Gs Kwon
- Kang