• Journal of Ginseng Research
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• v.45 no.2
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• pp.246-253
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• 2021

Background: Ginseng is one of the most valuable herbal supplements. It is challenging to perform quality control of ginseng products due to the diversity of bioactive saponins in their composition. Acid or alkaline hydrolysis is often used for the structural elucidation of these saponins and sugars in their side chains. Complete transformation of the original ginsenosides into their aglycones during the hydrolysis is one of the ways to determine a total saponin group content. The main hurdle of this approach is the formation of various by-products that was reported by many authors. Methods: Separate HPLC assessment of the total protopanaxadiol, protopanaxatriol and ocotillol ginsenoside contents is a viable alternative to the determination of characteristic biomarkers of these saponin groups, such as ginsenoside Rf and pseudoginsenoside F₁₁, which are commonly used for authentication of P. ginseng Meyer and P. quinquefolius L. samples respectively. Moreover, total ginsenoside content is an ideal aggregated parameter for standardization and quality control of ginseng-based medicines, because it can be directly applied for saponin dosage calculation. Results: Different hydrolysis conditions were tested to develop accurate quantification method for the elucidation of total ginsenoside contents in herbal products. Linearity, limits of quantification, limits of detection, accuracy and precision were evaluated for the developed HPLC-MS method. Conclusion: Alkaline hydrolysis results in fewer by-products than sugar elimination in acidic conditions. An equimolar response, as a key parameter for quantification, was established for several major ginsenosides. The developed approach has shown acceptable results in the analysis of several different herbal products.

• Journal of Ginseng Research
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• v.23 no.4
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• pp.230-234
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• 1999

Potassium channels play an important role in regulating vascular smooth muscle tone. Four types of $K^+$ channels areknown to be expressed in vascular smooth muscle cells, and maxi $Ca^{2+}-activated\;K^+$ channel $(BK_{Ca})$ is a dominant type of $K^+$ channels in these cells. Because total ginseng saponins and ginsenoside $Rg_3$ cause vasodilation with unclear mechanisms, we hypothesized that total ginseng saponins and ginsenoside $Rg_3$ induce vasodilation via activation of maxi $Ca^{2+}-activated\;K+$ channels. Whole-cell BKe. currents were voltage-dependent with half maximum activation at -14 mV, and the currents were sensitive to nanomolar ChTX and millimolar TEA. External application of total ginseng saponins increased the anlplitude of the whole-cell BKe. current in a concentration-dependent manner. Single-channel analysis indicates that total ginseng saponins caused the channel opening for a longer period of time. Ginsenoside $Rg_3$ increased the amplitude of whole-cell $K_{Ca}$ currents without affecting voltage dependence of the currents and increased single-channel open time. Hence, the results suggest that ginseng saponin-induced vasodilation may be due to activation of $K_{Ca}$.

• Journal of Ginseng Research
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• v.22 no.3
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• pp.205-210
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• 1998

Glucosidic bonds at the C20 position of the sapogenins were hydrolyzed easily in the lower pH, higher temperatures and longer times to give prosapogenins and sugars. The glucosidic bond of saponin at the C3 of ginsenoside-Rb1, which is secondary carbon, was relatively stable due to the low electron density of -0.2. But the bond of saponin at the C20 position, which is tertiary carbon with the relatively high electron density of -0.3, was liable to be hydrolyzed even in weakly acidic solution by the increase of heating time. On the other hand, red ginseng contained 13.34 mg/g of citric acid, 8.78 mg/g of malonic acid, 3.70 mg/g of oxalic acid, 2.13 mg/g of malic acid and 0.44 mg/g of succinct acid. Ginseng saponins were very stable in ginseng extract neutralized with sodium carbonate or sodium bicarbonate corresponding to the equivalent amount of the total organic acid in the red ginseng.

• Journal of Ginseng Research
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• v.20 no.3
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• pp.274-283
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• 1996

In this study, the effects of red ginseng components [ginsenosides (total saponins and $Rg_1$) on the function of ryanodine receptor (RyR) -$Ca^{2+}$ release channel complex protein (named as RyR or $Ca^{2+}$ channel), a membrane protein in sarcoplasmic reticulum (SR) of rabbit skeletal muscle were examined at the SR vesicle's level and the molecular levels with Chaps-solubilized and purified $Ca^{2+}$ channel protein and with reconstituted proteoliposomes by dialysis. The results were as follows. 1. The binding of ryanodine known as inhibitor of muscle contraction to the RyR was decreased at the whole range of concentration ($10^2$~$10^7$%) by these two ginseng components. In heavy SR vesicles, Chaps-solubilized and purified $Ca^{2+}$ channel protein, and reconstituted vesicles, its maximal inhibition by total saponins was shown at the concentration of $10^3$, $10^3$%, and $10^5$% respectively, and by gin- senoside $Rg_1}$) each was $10^3$%, $10^3$%, and $10^4$%. 2. The release of $Ca^{2+}$ ion through $Ca^{2+}$ channel in heavy SR vesicles and reconstituted proteoliposomes was increased as a whole by these two ginseng components, and particularly maximal release by both of them was shown at the range of $10^4$~$10^6$%. These results were seemed to be caused by conformational change of $Ca^{2+}$ release channel protein (RyR) by red ginseng components [ginsenosides (total saponins and $Rg_1}$).

• Journal of Ginseng Research
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• v.24 no.4
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• pp.188-195
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• 2000

Using Ginseng saponins (crude saponin and total saponin) and ginsenoside Rbl Rb2, Rc, Rd, Re, Rhl, and Rh2 in this study, we have examined the effects of the compounds on the induction of differentiation in normal rat mammary epithelial cells and 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor cells in culture. When normal rat mammary organoids were cultured in 100-mm culture plates in the presence or absence of ginseng saponins, there were four different cell colonies after two weeks in culture: cobble stone, spindle, honey comb, and senescence type colonies. Ginseng saponins showed different effects on the development of each colonies. Scrape-loading dye transfer tech-nique was performed to measure the effects of total saponin, Rhl, and Rh2 on intercellular junctional communication. Intercellular communication was not observed at short cultilral time, e.g., four or seven days, but when it cultured it up to two weeks, cell to cell communication was observed in saponin-treated cells. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from normal mammary organoids (e.g., ductal, webbed, stellate, and squamous colonies) or DMBA-induced mammary tumor (e.g., alveolar unit, foamy alveolar unit, squamous metaplasia, lobule-ductal, stellate, and webbed colony). In ginseng saponin-treated groups, webbed colonies were more and squamous colonies were less than control group. Moreover, the ductal colonies, marker tructure of well-differentiate mammary epithelial cells, were developed more in saponin-treated group than in control group. In conclusion, ginseng saponins affected on the differentiation of normal rat mammary epithelial cells and DMBA-induced mammary tumor cells in culture.

• Journal of Ginseng Research
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• v.19 no.3
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• pp.202-205
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• 1995

We have previously reported that the antagonism of U-50,488H-induced antinociception in mice pretreated with ginseng total saponins (GTS) Ivas abolished by pretreatment with a serotonin precursor, 5-hydroxytryptophan (5-HTP), but not by a noradrenaline precursor, L-dihydroxyphenylalanine (L-DOPA) in the tail flick test. In the present experiments, the effect of the same GTS on the development of tolerance to U-50,488H-induced antinociception was determined. GTS inhibited the development of tolerance to U-50,488H-induced antinociception. The inhibitory effect of GTS on the development of tolerance to U-50,488H-induced antinociception was reversed by 5-HTP, but not by L-DOPA. These findings suggest that the inhibitory effect of GTS on the development of tolerance to U-50,488H-induced antinociception is dependent on serotonergic mechanisms. Key words Ginseng total saponin, U-50,488H, tolerance, serotonin.

• Archives of Pharmacal Research
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• v.16 no.3
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• pp.237-243
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• 1993

These studies were performed to investigate the acting sites of ginseng total saponins (GTS) on the U-50, 488H-induced antinociception and the inhibitory effect of the development of tolerance to U-50, 488H-induced antinociception by GTS were studied. The U-50, 488H-induced antinociception was ntagonized in mice pretreated with GTS intraperitoneally, intracerebrally. These antagonisms were reversed by the pretratment iwth a serotonin precursor, 5-hydroxytrypophan (5-HTP), but not with a noradrenaline precursor, L-dihydroxyphenylalanine (L-DOPA). However, the intraplantar sites. On the other hand, GTS inhibited the development of tolerance to U-50, 488H-induced antinociception was reversed by pretreatment with 5-HTP, but not with L-DOPA. Therefore, the antagonism of U-50, 488H-induced antinociception and the inhibition of the development of tolerance to U-50, 488H-induced antinociception and the inhibition of the development of tolerance to U-50, 488H-induced antinociception by GTS are dependent on serotonegic mechanisms.

• YAKHAK HOEJI
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• v.29 no.2
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• pp.76-89
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• 1985

The effects of ginseng saponin on the activity, phosphorylation, [$^{3}$H] ouabain binding and light scattering (disruption) of purified $Na^{+}$ ,$K^{+}$ -ATPase isolated from the outer medulla of sheep kidney were compared to those of gypsophila saponin, sodium dodecylsulfate (SDS), and Triton X-100 on the same parameters. $Na^{+}$ , $K^{+}$ -ATPase activity, phosphorylation, and [$^{3}H$] ouabain binding were inhibited by ginseng saponin (triol>total>diol), SDS, or Triton X-100, but increased by gypsophila saponin. Low doses of ginseng saponin (3.mu.g saponin/.mu.g protein) decreased phosphorylation sites and ouabain binding site concentration (Bmax) without any change of turnover number and affinity for ouabain binding which were decreased by high dose of ginseng saponin (over 10.mu.g saponin/.mu.g protein), SDS or Triton X-100. On the other hand, gypsophila saponin increased the affinity without any change of Bmax for ouabain binding. Inhibition of $Na^{+}$ ,$K^{+}$ -ATPase activity by ginseng saponin and SDS or Triton X-100 appeared before and after decrease in light scattering, respectively. These data suggest that ginseng saponins (total, diol, triol saponin) inhibit $Na^{+}$ , $K^{+}$ -ATPase activity by specific direct and general detergent action at low and high concentrations, respectively, and this inhibitory action of ginseng sapornin to $Na^{+}$ , $K^{+}$ -ATPase is not general action of all saponins.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.116-123
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• 1998

고양이 말초혈액 탐식세포(단핵구세포(MNC) 및 다형핵백혈구(PMNC))의 탐식능 에 있어서 인삼 saponin(ginseng total saponin(G75), ginseng PT saponin(GPT) 및 ginseng PD saponin(GPD))의 면역증강 효과를 flow cytometry를 이용하여 분석하였다. 인삼 ssponins을 직접 첨가하여 배양한 MNC 및 PMNC에서는 탐식중강 효과가 나타나지 않았다. 각각의 인삼 saponin을 첨가하여 배양한 PMNC 및 MNC 배양상충액의 존재하에 PMNC 및 MNC의 탐식능을 겅토한 결과, MNC의 탐식능은 Gff 첨가 PMNC 배양상충액과 GTS 및 GPT 첨가 MNC 배양상충액의 존재하에서 약간의 탐식증강 효과를 보였다. PMNC 탐식능의 경우에는 GPD 첨가 PMNC 배양상충액에서 미약한 탐식증강 효과가 나타났으나, 각각의 인 삼 saponin 첨가 MNC 배양상충액 존재하에서는 모두 현저한 탐식중강 효과를 나타내었다. 이상의 결과로부터 고양이 말초혈액 탐식세포의 탐식증강 효과는 인삼 saponin의 직접적인 작용보다는 인삼 saponin에 의해 활성화된 단핵구세포에서 분비되는 가용성물질에 의해 단 핵구세포보다는 다형핵백혈구에서 현저하게 탐식효과가 증강되는 것으로 판단되었다.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.366-374
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• 2016

Background: In this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09. Methods: The transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis. Results: A total of 32 ${\beta}$-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$; $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$; $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$; $Rg1{\rightarrow}Rh1$. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively. Conclusion: This is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.