Fermented Ginseng Extract Research Articles

Fermented black ginseng extract prevents UVB-induced inflammation by regulating the nc886-PKR pathway in human keratinocytes.

Continuous exposure of the skin to ultraviolet B (UVB) rays can cause inflammation and photodamage. In previous studies, we observed that the upregulation of nc886, a noncoding RNA (ncRNA), can alleviate UVB-induced inflammation through suppression of the protein kinase RNA (PKR) pathway. We aim to investigate the effect of fermented black ginseng extract (FBGE), which has been shown to increase the expression of nc886, on UVB-induced inflammation in keratinocytes. To confirm the cytotoxicity of FBGE, MTT assay was performed, and no significant cytotoxicity was found on human keratinocytes. The efficacies of FBGE were assessed through qPCR, Western blotting, and ELISA analysis which confirmed regulation of UVB-induced inflammation. The analysis results showed that FBGE inhibited the decrease in nc886 expression and the increase in the methylated nc886 caused by UVB. It also prevented the UVB-induced increase of metalloproteinase-9 (MMP-9), metalloproteinase-1 (MMP-1), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). Additionally, FBGE suppressed the PKR-MAPK pathways activated by UVB. These results implicate that FBGE can alleviate UVB-induced inflammation through regulation of the nc886-PKR pathway.

Ginsenoside compound K-loaded gold nanoparticles synthesized from Curtobacterium proimmune K3 exerts anti-gastric cancer effect via promoting PI3K/Akt-mediated apoptosis

BackgroundCompound K (CK) is the minor ginsenoside present in fermented Panax ginseng extract. Despite the pharmacological efficacy of CK, its industrial use has been restricted due to its low water solubility and poor permeability. To overcome this defect, our study was to synthesize gold nanoparticles from CK (CK-AuNPs) to investigate their potential as anticancer candidates.MethodsTo biologically synthesize CK-AuNPs, a novel strain, Curtobacterium proimmune K3, was isolated from fermented ginseng beverage, then combined with CK and gold salts to biosynthesize gold nanoparticles (CurtoCK-AuNPs). Their physicochemical characteristics were evaluated using UV–Vis spectrometry, FE-TEM, EDX, elemental mapping, XRD, SAED, DLS and TGA.ResultsCurtoCK-AuNPs exerted significant selective cytotoxic effects on AGS human gastric cancer cells. Fluorescence staining with Hoechst, propidium iodide, and MitoTracker demonstrated that CurtoCK-AuNPs induce apoptosis and mitochondrial damage, respectively. Quantitative real-time PCR and western blotting analyses showed that cytotoxic effect of CurtoCK-AuNPs were involved in apoptosis, based on their activation of Bax/Bcl-2, cytochrome c, caspase 9, and caspase 3, as well as their suppression of PI3K–Akt signaling.ConclusionOur findings provide data for understanding the molecular mechanisms of nanoparticles; thus, providing insight into the development of alternative medications based on gold nanoparticles of ginseng-derived CK.

Antioxidant Activity and Inhibitory Effect on Nitric Oxide Production of Hydroponic Ginseng Fermented with Lactococcus lactis KC24.

Panax ginseng Meyer is used as a medicinal plant. The aim of this study was to ferment hydroponic ginseng with Lactococcus lactis KC24 and confirm its antioxidant activity and inhibitory effect on nitric oxide (NO) production. Flavonoid and phenol contents in fermented ginseng extracts were measured. Antioxidant activity was measured by DPPH, ABTS, reducing power, FRAP and β-carotene assays. Additionally, inhibitory effects on NO production and toxicity of the fermented extract were determined using RAW 264.7 cells. Phenol and flavonoid contents increased as the fermentation time increased, and the contents were higher in hydroponic ginseng than in soil-cultivated ginseng. The DPPH assay revealed that the antioxidant activity of the 24 h fermented extract significantly increased from 32.57% to 41% (p < 0.05). The increase in antioxidant activity may be affected by an increase in phenol and flavonoid contents. At 1 mg/mL solid content, the 24 h fermented hydroponic ginseng extract inhibited NO production from 9.87 ± 0.06 μM to 1.62 ± 0.26 μM. In conclusion, the increase in antioxidant activity affects the inhibition of NO production, suggesting that fermented hydroponic ginseng may be used in the industries of functional food and pharmaceutical industry as a functional material with anti-inflammatory effects.

Fermented Ginseng Extract, BST204, Suppresses Tumorigenesis and Migration of Embryonic Carcinoma through Inhibition of Cancer Stem Cell Properties.

The pharmacological effects of BST204—a fermented ginseng extract—on several types of cancers have been reported. However, the effects of ginseng products or single ginsenosides against cancer stem cells are still poorly understood. In this study, we identified the anti-tumorigenic and anti-invasive activities of BST204 through the suppression of the cancer stem cell marker, CD133. The treatment of embryonic carcinoma cells with BST204 induced the expression of the tumor suppressor protein, p53, which decreased the expression of cell cycle regulatory proteins and downregulated the expression of CD133 and several stemness transcription factors. These changes resulted in both the inhibition of tumor cell proliferation and tumorigenesis. The knockdown of CD133 suggests that it has a role in tumorigenesis, but not in cancer cell proliferation or cell cycle arrest. Treatment with BST204 resulted in the reduced expression of the mesenchymal marker, N-cadherin, and the increased expression of the epithelial marker, E-cadherin, leading to the suppression of tumor cell migration and invasion. The knockdown of CD133 also exhibited an anti-invasive effect, indicating the role of CD133 in tumor invasion. The single ginsenosides Rg3 and Rh2—major components of BST204—exhibited limited effects against cancer stem cells compared to BST204, suggesting possible synergism among several ginsenoside compounds.

Fermented Wild Ginseng by Rhizopus oligosporus Improved l-Carnitine and Ginsenoside Contents.

We conducted this study to investigate the beneficial effects of Rhizopus oligosporus fermentation of wild ginseng on ginsenosides, l-carnitine contents and its biological activity. The Rhizopus oligosporus fermentation of wild ginseng was carried out at 30 °C for between 1 and 14 days. Fourteen ginsenosides and l-carnitine were analyzed in the fermented wild ginseng by the ultra high pressure liquid chromatography–mass spectrometry (UPLC–MS) system. Our results showed that the total amount of ginsenosides in ginseng increased from 3274 to 5573 mg/kg after 14 days of fermentation. Among the 14 ginsenosides tested, the amounts of 13 ginsenosides (Rg1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg2, Rg3, Rh1, compound K, F1 and F2) increased, whereas ginsenoside Rb1 decreased, during the fermentation. Furthermore, l-carnitine (630 mg/kg) was newly synthesized in fermented ginseng extract after 14 days. In addition, both total phenol contents and DPPH radical scavenging activities showed an increase in the fermented ginseng with respect to non-fermented ginseng. These results show that the fermentation process reduced the cytotoxicity of wild ginseng against RAW264.7 cells. Both wild and fermented wild ginseng showed anti-inflammatory activity via inhibition of nitric oxide synthesis in RAW264.7 murine macrophage cells.

Anti-dermatitic effect of fermented ginseng extract including rich compound K through inhibiting activation of macrophage.

The purpose of this study is to investigate the effect of fermented ginseng extract by Lactobacillus brevis (FGE) on lipopolysaccharide (LPS)-activated macrophages and 12-O-tetradecanoylphorbol-13-acetate (TPA)-mediated dermatitis in mice. FGE showed better anti-inflammatory activities than ginseng extract on the formation of nitric monooxide, IL-6, TNF-α, and IL-10 within non-cytotoxicity range in LPS-activated RAW264.7 cells. In addition, FGE reduced the expression of cyclooxygenase-2 and inducible nitric oxide synthase through inhibiting nuclear translocation of NF-κB. Consistent with in vitro experiments, FGE dose-dependently suppressed ear edema, and formation of TNF-α and IL-6, and it (50mg/mL) significantly enhanced IL-10 level in ear tissues of TPA-treated mice. In conclusions, FGE has anti-dermatitic activity through inhibiting the activation of macrophages. Such effects of FGE are associated with suppressing nuclear translocation of NF-κB. Therefore, the features of FGE may provide the information for its application for therapy and prevention of dermatitis.

Skin lightening effect of fermented Panax ginseng extract

Skin lightening effect of fermented Panax ginseng extract

Enhanced production of compound K in fermented ginseng extracts by Lactobacillus brevis.

The purpose of this study is to establish the best condition and microorganism for preparation of fermented ginseng including rich compound K. When raw ginseng parts were incubated with various microorganisms, there was an increase in compound K at 5days in all samples fermented by Lactobacillus brevis (L. brevis) and Lactobacillus plantarum, isolated from kimchi. Especially, ginseng fine roots fermented with L. brevis (FR-B) included higher levels of compound K, total phenolic compounds, and antioxidant activities compared with other products. Conclusionally, these results indicate that the optimum condition for providing rich compound K product in fermented ginseng is ginseng fine roots are fermented with L. brevis for 5days. Additionally, with FR-B there was greater improvement in physiochemical properties than with other products. Such information may be helpful for the manufacture of fermented ginseng including rich compound K as well as for understanding the biological features of fermented ginseng.

Antiviral Activity of Fermented Ginseng Extracts against a Broad Range of Influenza Viruses.

Ginseng products used as herb nutritional supplements are orally consumed and fermented to ginsenoside compounds by the intestinal microbes. In this study, we investigated antiviral protective effects of fermented ginseng extracts against different strains of influenza viruses in genetically diverse mouse models. Intranasal coinoculation of mice with fermented ginseng extract and influenza virus improved survival rates and conferred protection against H1N1, H3N2, H5N1, and H7N9 strains, with the efficacy dependent on the dose of ginseng samples. Antiviral protection by fermented ginseng extract was observed in different genetic backgrounds of mice and in the deficient conditions of key adaptive immune components (CD4, CD8, B cell, MHCII). The mice that survived primary virus inoculation with fermented ginseng extract developed immunity against the secondary infection with homologous and heterosubtypic viruses. In vitro cell culture experiments showed moderate virus neutralizing activity by fermented ginseng extract, probably by inhibiting hemagglutination and neuraminidase activity. This study suggests that fermented ginseng extracts might provide a means to treat influenza disease regardless of virus strains.

Fermentation-Mediated Enhancement of Ginseng's Anti-Allergic Activity against IgE-Mediated Passive Cutaneous Anaphylaxis In Vivo and In Vitro.

Ginseng (the root of Panax ginseng Meyer) fermented by Lactobacillus plantarum has been found to attenuate allergic responses in in vitro and in vivo experimental models. Ginseng has been reported to also possess various biological functions including anti-inflammatory activity. The present study was aimed at comparing the anti-allergic effect of ginseng and fermented ginseng extracts on IgE-mediated passive cutaneous anaphylaxis in vitro in a murine cell line and in vivo in mice. Fermented ginseng extract (FPG) showed higher inhibitory effect against in vitro and in vivo allergic responses when compared with ginseng extract (PG). The secretion of β-hexosaminidase and interleukin (IL)-4 from the IgE-DNP-stimulated RBH-2H3 mast cells were significantly (p < 0.05) inhibited by FPG treatment, and this effect was concentration-dependent. Further, MKK4 activation and subsequent JNK phosphorylation were attenuated by FPG treatment. The inhibitory effect of FPG on the in vitro allergic response was verified in vivo against IgE-DNP-induced passive cutaneous anaphylaxis in a mouse model. These data indicated that the fermentation of ginseng with L. plantarum enhanced its anti-allergic effects both in vitro and in vivo. We predict that compositional changes in the ginsenosides caused by the fermentation may contribute to the change in the anti-allergic effects of ginseng. The results of our study highlight the potential of the use of FPG as a potential anti-allergic agent.

Fermented ginseng extract, BST204, disturbs adipogenesis of mesenchymal stem cells through inhibition of S6 kinase 1 signaling

BackgroundThe biological and pharmacological effects of BST204, a fermented ginseng extract, have been reported in various disease conditions. However, its molecular action in metabolic disease remains poorly understood. In this study, we identified the antiadipogenic activity of BST204 resulting from its inhibition of the S6 kinase 1 (S6K1) signaling pathway. MethodsThe inhibitory effects of BST204 on S6K1 signaling were investigated by immunoblot, nuclear fractionation, immunoprecipitation analyses. The antiadipogenic effect of BST204 was evaluated by measuring mRNA levels of adipogenic genes and by chromatin immunoprecipitation and quantitative real-time polymerase chain reaction analysis. ResultsTreatment with BST204 inhibited activation and nuclear translocation of S6K1, further decreasing the interaction between S6K1 and histone H2B in 10T1/2 mesenchymal stem cells. Subsequently, phosphorylation of H2B at serine 36 (H2BS36p) by S6K1 was reduced by BST204, inducing an increase in the mRNA expression of Wnt6, Wnt10a, and Wnt10b, which disturbed adipogenic differentiation and promoted myogenic and early osteogenic gene expression. Consistently, BST204 treatment during adipogenic commitment suppressed the expression of adipogenic marker genes and lipid drop formation. ConclusionOur results indicate that BST204 blocks adipogenesis of mesenchymal stem cells through the inhibition of S6K1-mediated histone phosphorylation. This study suggests the potential therapeutic strategy using BST204 to combat obesity and musculoskeletal diseases.

Anti‐inflammatory and cytotoxic effects of ginseng extract bioconverted by Leuconostoc mesenteroides KCCM 12010P isolated from kimchi

SummaryA bioconversion technique using microorganisms has been applied to ginseng to increase content of bioactive ginsenoside and biofunctionality such as anticancer, anti‐obesity and antioxidant activities. The objective of this study was to screen lactic acid bacteria for bioconversion of ginsenosides and to evaluate anti‐inflammatory and cytotoxic effects of bioconverted ginseng extract. Strains isolated from kimchi were screened for their β‐glucosidase activities using esculin agar. Selected strain was identified based on 16S rRNA sequencing and carbohydrate fermentation. During ginseng fermentation, viable cell number and pH were determined. Bioconverted ginsenosides were analysed by HPLC. Anti‐inflammatory effects were evaluated using RAW 264.7 cells, and cytotoxic effects were determined by MTT assay. Among 166 isolates screened, Leuconostoc mesenteroides was selected for ginseng bioconversion, as it showed a higher β‐glucosidase activity and viable cell number than any of the other tested strains. After fermentation for 2 days, viable cell number was 8.8 log CFU mL−1 and final pH was 4.8. Ginsenoside Rb2 was bioconverted into ginsenoside Rg3 (Rb2 → Rd → Rg3) by L. mesenteroides. The nitric oxide contents of 2‐day‐fermented extract decreased by as much as 25%, compared to a non‐fermented extract. The cell viabilities of HepG2, HT‐29, HeLa and LoVo treated with fermented ginseng extract also decreased by 49.7%, 20.2%, 21.0% and 8.7%, respectively, compared to those of control cells treated with non‐fermented extract. Ginseng extract bioconverted by L. mesenteroides showed anti‐inflammatory and anticancer effects. Therefore, bioconverted ginseng extract might have applications in the pharmaceutical and/or functional food industry.

Protective effects of cultured and fermented ginseng extracts against scopolamine-induced memory loss in a mouse model

This study was performed to investigate the effect of a concentrate of fermented wild ginseng root culture (HLJC0701) on memory improvement in the scopolamine (SPL)-induced memory-deficient mouse model. Eight-week-old male ICR mice were used to evaluate the protective effect of HLJC0701 against the SPL-induced memory loss animal model. The Morris water maze test, which measures hippocampus-dependent learning ability, and the Y-maze test, a short-term memory assessment test, were performed and related markers were analyzed. HLJG0701-treated groups displayed significantly reduced acetylcholinesterase activity and increased acetylcholine level compared with the SPL-administered group (SPL-G) (P<0.05). In the Y-maze test, the spontaneous alternation in al HLJC0711-treated groups was significantly increased compared with that in SPL-G (P<0.05). In the Morris water maze test, the escape latency and time spent in the target quadrant in all HLJC0701-treated groups were significantly decreased and increased, respectively, compared with those in SPL-G (P<0.05). In addition, the brain-derived neurotrophic factor level in groups treated with HLJC0701 300 and 600 mg/kg body weight was significantly increased compared with that in SPL-G (P<0.05). These results suggest that the HLJC0701 may protect against memory loss by inhibiting acetylcholinesterase activity and preventing acetylcholine deficiency.

Protective effects of cultured and fermented ginseng extracts against scopolamine-induced memory loss in a mouse model

[This retracts the article on p. 37 in vol. 34, PMID: 29628975.].

Ginsenoyne C, a polyacetylene isolated from Panax Ginseng inhibit inflammatory mediators via regulating extracellular regulated kinases signaling

Background: Panax ginseng has been used as a traditional medicine for various pathological conditions. Ginsenoyne C (GSC) is a polyacetylene, minor constituent. Objective: The objective of the present study was to determine the anti-inflammatory activities of GSC for the protection of inflammation. Materials and Methods: The effects of GSC on cell viability and nitric oxide (NO) production in lipopolysaccharide (LPS)–induced RAW 264.7 cells were investigated through MTT assay and the Griess reaction, respectively. The levels of inflammatory cytokines such as interleukin (IL-1β) and IL-6 and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and pERK were measured using enzyme-linked immunosorbent assay and Western blot analysis, respectively. Results: GSC suppressed NO production in LPS-induced RAW 264.7 cells. GSC suppressed the production of inflammatory mediators through downregulating phosphorylation of extracellular regulated kinases signaling in LPS-induced RAW 264.7. Conclusion: GSC has a potential therapeutic agent for protection of inflammation. Abbreviation used: CCK: Cell counting kit; COX-2: Cyclooxygenase-2; ELISA: Enzyme linked-immunosorbent assay; ERK: Extracellular regulated kinases; FBS: Fetal bovine serum; FEG: Fermented ginseng extract; GSC: Ginsenoyne C; IL: Interleukin; iNOS: Inducible nitric oxide synthase; LPS: Lipopolysaccharide; MAPK: Mitogen-activated protein kinases; NO: Nitric oxide; PVDF: Polyvinylidene difluoride; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Ginseng fermented by mycotoxin non-producing Aspergillus niger: ginsenoside analysis and anti-proliferative effects.

Korean ginseng was fermented using Aspergillus niger (A. niger) FMB S494 and mycotoxins such as ochratoxin and fumonisin were not detected in the fermented ginseng. Protopanaxadiol-type ginsenosides such as glycosylated forms of Rb1, Rb2, Rc, and Rd decreased to 0 while compound K (cK) increased from 0 to 9×104ppm in the extract of fermented ginseng. Protopanaxtriol-type ginsenosides such as Re and Rg1 decreased from 7.1×104 to 3.0×104ppm and 6.8×104 to 4.6×104ppm, respectively. Rg2 and Rh1 increased from 0 to 1.9×104ppm and 0 to 2.7×104ppm, respectively. We can demonstrate that A. niger was more inclined to transform protopanaxadiol-type ginsenosides. Moreover, fermented ginseng extract showed a dramatically enhanced anti-proliferative effect on human HT-29 cell line with a minimum effective concentration of about 1µg/mL, which might be attributed to the high degree of biotransformation of ginsenosides, especially the high output of ginsenoside cK.

Metabolite profiling of fermented ginseng extracts by gas chromatography mass spectrometry

BackgroundGinseng contains many small metabolites such as amino acids, fatty acids, carbohydrates, and ginsenosides. However, little is known about the relationships between microorganisms and metabolites during the entire ginseng fermentation process. We investigated metabolic changes during ginseng fermentation according to the inoculation of food-compatible microorganisms. MethodsGas chromatography mass spectrometry (GC-MS) datasets coupled with the multivariate statistical method for the purpose of latent-information extraction and sample classification were used for the evaluation of ginseng fermentation. Four different starter cultures (Saccharomyces bayanus, Bacillus subtilis, Lactobacillus plantarum, and Leuconostoc mesenteroide) were used for the ginseng extract fermentation. ResultsThe principal component analysis score plot and heat map showed a clear separation between ginseng extracts fermented with S. bayanus and other strains. The highest levels of fructose, maltose, and galactose in the ginseng extracts were found in ginseng extracts fermented with B. subtilis. The levels of succinic acid and malic acid in the ginseng extract fermented with S. bayanus as well as the levels of lactic acid, malonic acid, and hydroxypruvic acid in the ginseng extract fermented with lactic acid bacteria (L. plantarum and L. mesenteroide) were the highest. In the results of taste features analysis using an electronic tongue, the ginseng extracts fermented with lactic acid bacteria were significantly distinguished from other groups by a high index of sour taste probably due to high lactic acid contents. ConclusionThese results suggest that a metabolomics approach based on GC-MS can be a useful tool to understand ginseng fermentation and evaluate the fermentative characteristics of starter cultures.

Fermentative transformation of ginsenoside Rb1 from Panax ginseng C. A. Meyer to Rg3 and Rh2 by Lactobacillus paracasei subsp. tolerans MJM60396

Lactic acid bacteria (LAB) were screened for ginsenoside transforming activity using crude ginseng extract. Thin-layer chromatography analysis of fermented ginseng extract showed that LAB strain MJM60396 possessed higher ginsenoside transformation ability than other strains. It converted major ginsenosides into minor ginsenosides such as Rg3 and Rh2. MJM60396 also showed high β-glucosidase activity. Strain MJM60396 was identified as Lactobacillus paracasei subsp. tolerans based on 16S rRNA gene sequence. To delineate the pathway involved in the production of the minor ginsenosides Rg3 and Rh2, strain MJM60396 was incubated with pure ginsenoside Rb1. HPLC analysis revealed the appearance of Rg3 and Rh2 peak from the incubation mixture containing Rb1 and strain MJM60396. Furthermore, β-glucosidase enzyme was prepared from strain MJM60396. To achieve its maximum activity, we optimized the pH and temperature conditions. Cell-free β-glucosidase enzyme hydrolyzed ginsenoside Rb1 through the following pathway: ginsenoside Rb1 → Rd → Rg3 → Rh2. This is the first report on the transformation of ginsenosides Rb1 to Rg3 and Rh2 by a Lac. paracasei subsp. tolerans strain. Our results indicate that Lac. paracasei subsp. tolerans MJM60396 has the potential to be used for preparing ginsenosides Rg3 and Rh2 as nutraceuticals.

Isolation and identification of antiproliferative substances from ginseng fermented using Ganoderma lucidum mycelia

Two active substances from ginseng fermented using Ganoderma lucidum mycelia were investigated for antiproliferative effects against the human lung cancer cell line A549. The chloroform fraction of fermented ginseng extracts showed a strong antiproliferative effect. This fraction was isolated and purified using silica gel and C18 resin column chromatography and semi-preparative reverse phase HPLC. The structures of isolated compounds were determined using spectroscopic methods (ESI-MS, 1H and 13C NMR). Isolated compounds were identified as ginsenoside compound K and 3-oxo-compound K. Both inhibited A549 cell growth in a dose-dependent manner. Cell viability values for ginsenoside compound K were 74.88, 59.30, 5.76, 5.79, and 6.27% at 6.25, 12.50, 25.00, 50.00, and 100.00 μg/mL, respectively, and ginsenoside 3-oxo-compound K showed values 89.40, 59.62, 6.05, and 4.64% at 3.70, 7.50, 15.00, and 30.00 μg/mL, respectively. Compound K and 3-oxo-compound K from fermented ginseng can be used as natural anti-cancer agents.

Stereoselective determination of ginsenosides Rg3 and Rh2 epimers in rat plasma by LC‐MS/MS: Application to a pharmacokinetic study

We developed and validated an accurate and sensitive LC-MS/MS method for the simultaneous quantitation of ginsenoside Rg3 and Rh2 epimers (R-Rg3, S-Rg3, R-Rh2, and S-Rh2) in rat plasma. Analytes were extracted from 0.1 mL aliquots of rat plasma by liquid-liquid extraction, using 2 mL of ethyl acetate. In this assay, dioscin (500 ng/mL) was used as an internal standard. Chromatographic separation was conducted using an Acclaim RSLC C18 column (150 × 2.1 mm, 2.2 μm) at 40°C, with a gradient mobile phase consisting of 0.1% formic acid in distilled water and in acetonitrile, a flow rate of 0.35 mL/min, and a total run time of 20 min. Detection and quantification were performed using a mass spectrometer in selected reaction-monitoring mode with negative electrospray ionization at m/z 783.4 → 161.1 for R-Rg3 and S-Rg3, m/z 621.3 → 161.1 for R-Rh2 and S-Rh2, and m/z 867.2 → 761.5 for the internal standard. For R-Rg3 and S-Rg3, the lower limit of quantification was 5 ng/mL, with a linear range up to 500 ng/mL; for R-Rh2 and S-Rh2, the lower limit of quantification was 150 ng/mL, with a linear range up to 6000 ng/mL. The coefficient of variation for assay precision was less than 10.5%, with an accuracy of 86.4-112%. No relevant cross-talk or matrix effect was observed. The method was successfully applied to a pharmacokinetic study after oral administration of 400 mg/kg and 2000 mg/kg of BST204, a fermented ginseng extract, to rats. We found that the S epimers exhibited significantly higher plasma concentrations and area under curve values for both Rg3 and Rh2. This is the first report on the separation and simultaneous quantification of R-Rg3, S-Rg3, R-Rh2, and S-Rh2 in rat plasma by LC-MS/MS. The method should be useful in the clinical use of ginseng or its derivatives.