Article

Simultaneous Determination of Nine Major Flavonoids in Sophora flavescens by RP-LC

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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.

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... 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. ...
Article
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]. ...
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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]. ...
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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.
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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.
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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.
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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.
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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.
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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