Simplified Extraction of Ginsenosides from American Ginseng (<i>Panax quinquefolius </i>L.) for High-Performance Liquid Chromatography−Ultraviolet Analysis

Corbit, Rebecca; Ferreira, Jorge; Ebbs, Stephen D.; Murphy, Laura

doi:10.1021/jf051504p

Cited by 74 publications

(38 citation statements)

References 48 publications

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“…Rg1 extraction by sonication in methanol has been proved to be an effective method reported by Chen et al (2000) and Corbit et al (2005). The modification in our report was that condensation of the condensation of Rg1 extract was achieved by nitrogen flowing at room temperature after sonication.…”

Section: Recoverymentioning

confidence: 77%

Transport behavior and efflux of Rg1 in rat pulmonary epithelial cells

Meng

Zhang

Zhao

et al. 2007

Biomedical Chromatography

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The aim of this study was to investigate whether ginsenoside Rg1 could be transported into rat pulmonary epithelial cells and its transport behavior and efflux through the cells. A high-performance liquid chromatography coupled with 2487 UV-vis detector at 203 nm was applied. The mobile phase was 0.05% phosphate-acetonitrile (75:25, v/v). Cells were incubated with Rg1 (100 microg/mL) for a specific time, then lysed and sonicated in methanol to extract intracellular Rg1. Cells incubated with Rg1 and verapamil or KCN were processed by the same method. A 20 microL aliquot of sample was injected into the HPLC system to determine Rg1 concentration. The results showed that Rg1 could be transported into the epithelial cells with peak concentration of 1.28 microg/10(5) cells at 0.5 h. Metabolic inhibitor KCN and P-glycoprotein inhibitor verapamil could increase Rg1 concentration within the cells, indicating that efflux of Rg1 was energy-dependent and P-gp was likely to be involved. This is the first time that the transport behavior and efflux of Rg1 through rat pulmonary epithelial cells has been demonstrated. The phenomenon that Rg1 concentration in the cells decreased whereas that in the medium remained high indicated that a more effective means of drug administration should be found.

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Section: Recoverymentioning

confidence: 77%

Transport behavior and efflux of Rg1 in rat pulmonary epithelial cells

Meng

Zhang

Zhao

et al. 2007

Biomedical Chromatography

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“…The estrogenic ginsenoside Rb1 is one of the most abundant ginsenosides found in methanol-extracted P quinquefolium root. 26,37 In the interpretation of results, it is also important to consider the influence of working with whole root extracts containing a mixture of approximately 20 ginsenosides that may have variable effects on cellular events depending on their individual concentrations, as well as cell culture conditions. At low concentrations of alc-GE, the net effect of the individually acting ginsenosides is an estrogenic response, possibly due to some estrogenic ginsenosides directly binding to ERs and others stimulating estrogenic responses independent of ER binding.…”

Section: Discussionmentioning

confidence: 99%

“…24,25 Most ginsenosides are soluble in aqueous alcohols, and alcohol-extracted ginseng contains greater ginsenoside concentrations than water onlyextracted ginseng does. 26 Thus, the processing method of the ginseng used in scientific study, as well as ginseng supplements, may account for reported differences in laboratory results. The goal of the present study was to determine the effects of alcohol-and water-extracted P quinquefolium root on human breast cancer cell proliferation, as well as to assess the involvement of estrogen receptors.…”

mentioning

confidence: 99%

Extraction-Dependent Effects of American Ginseng (Panax quinquefolium) on Human Breast Cancer Cell Proliferation and Estrogen Receptor Activation

2006

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Hypothesis:Ginseng root extracts and the biologically active ginsenosides have been shown to inhibit proliferation of human cancer cell lines, including breast cancer. However, there are conflicting data that suggest that ginseng extracts (GEs) may or may not have estrogenic action, which might be contraindicated in individuals with estrogen-dependent cancers. The current study was designed to address the hypothesis that the extraction method of American ginseng (Panax quinquefolium) root will dictate its ability to produce an estrogenic response using the estrogen receptor (ER)-positive MCF-7 human breast cancer cell model. Methods: MCF-7 cells were treated with a wide concentration range of either methanol-(alc-GE) or water-extracted (w-GE) ginseng root for 6 days. Cells were grown in media containing either normal or charcoal-stripped fetal calf serum to limit exposure to exogenous estrogen. Thus, an increase in MCF-7 cell proliferation by GE indicated potential estrogenicity. This was confirmed by blocking GE-induced MCF-7 cell proliferation with ER antagonists ICI 182,780 (1 nM) and 4-hydroxytamoxifen (0.1 µ µM). Furthermore, the ability of GE to bind ERα α or ERβ β and stimulate estrogen-responsive genes was examined. Results: Alc-GE, but not w-GE, was able to increase MCF-7 cell proliferation at low concentrations (5-100 µ µg/mL) when cells were maintained under low-estrogen conditions. The stimulatory effect of alc-GE on MCF-7 cell proliferation was blocked by the ER antagonists ICI 182,780 or 4-hydroxytamoxifen. At higher concentrations of GE, both extracts inhibited MCF-7 and ER-negative MDA-MB-231 cell proliferation regardless of media conditions. Binding assays demonstrated that alc-GE, but not w-GE, was able to bind ERα α and ERβ β. Alc-GE (50 µ µg/mL) also induced an approximate 2.5-fold increase in expression of the estrogenresponsive pS2 gene, as well as progesterone receptor (PgR) gene expression, whereas w-GE was without effect. Conclusion: These data indicate that low concentrations of alc-GE, but not w-GE, elicit estrogenic effects, as evidenced by increased MCF-7 cell proliferation, in a manner antagonized by ER antagonists, interactions of alc-GE with estrogen receptors, and increased expression of estrogenresponsive genes by alc-GE. Thus, discrepant results between different laboratories may be due to the type of GE being analyzed for estrogenic activity.

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“…The HWEG was tested using the E-TOXATE detection kit (Sigma-Aldrich; St. Louis, MO) to verify that bacterial endotoxin was not present. The final endotoxin-free HWEG was analyzed for presence and concentration of ginsenosides using high-performance liquid chromatography as reported elsewhere (Corbit et al, 2005). Lyophilized products were analyzed for ginsenoside content by HPLC (Corbit et al, 2005), total carbohydrate content by phenol-sulfuric method (Masuko et al, 2005), glycosyl composition by gas chromatography-mass spectrometry (King and Murphy, 2009), and protein content by BCA assay (Thermo Pierce; Rockford, IL).…”

Section: Ginseng Preparations Analysis and Dosingmentioning

confidence: 99%

“…The final endotoxin-free HWEG was analyzed for presence and concentration of ginsenosides using high-performance liquid chromatography as reported elsewhere (Corbit et al, 2005). Lyophilized products were analyzed for ginsenoside content by HPLC (Corbit et al, 2005), total carbohydrate content by phenol-sulfuric method (Masuko et al, 2005), glycosyl composition by gas chromatography-mass spectrometry (King and Murphy, 2009), and protein content by BCA assay (Thermo Pierce; Rockford, IL). The extract contained 6.89% (w/w) ginsenosides, a total carbohydrate content of 92% (w/w) (consisting of 99.3% glucose, 0.3% arabinose, 0.2% galactose, and 0.1% rhamnose), and a total protein content of less than 3.0%.…”

Section: Ginseng Preparations Analysis and Dosingmentioning

confidence: 99%