• Journal of Ginseng Research
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• v.38 no.2
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• pp.116-122
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• 2014

Ginseng roots were extracted with aqueous methanol, and extracts were suspended in water and extracted successively with ethyl acetate and n-butanol. Column chromatography using the n-butanol fraction yielded four purified triol ginseng saponins: the ginsenosides Re, Rf, Rg2, and 20-gluco-Rf. The physicochemical, spectroscopic, and chromatographic characteristics of the ginsenosides were measured and compared with reports from the literature. For spectroscopic analysis, two-dimensional nuclear magnetic resonance (NMR) methods such as $^1H$-$^1H$ correlation spectroscopy, nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond connectivity were employed to identify exact peak assignments. Some peak assignments for previously published $^1H$-and $^{13}C$-NMR spectra were found to be inaccurate. This study reports the complete NMR assignment of 20-gluco-Rf for the first time.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.29 no.4
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• pp.342-349
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• 1984

In order to obtain the basic information for the development of ginseng varieties with high saponin contents. saponin contents and ginsenosides of Panax ginseng (Korean ginseng) and Panax quinquefolium (American ginseng) grown under the same environmental conditions were analysed. Crude saponin contents of root and aerial parts were more in Panax quinquefolium than in Panax ginseng, and aerial parts had more saponin contents in comparison with a root. Protopanaxatriol saponin was greatly more in the aerial parts of ginseng while more amount of protopanaxadiol saponins were detected in the root. As for the ginsenosides, the patterns of ginsenosides detected in total saponin of the aerial parts were not different between two species, Panax ginseng and Panax quinquefolium, but the root ginsenoside patterns were quite different. Ginsenosides such as Rg$_2$, R$_{f}$. R$_{a}$ and R$_{o}$ were not detected in the root of Panax quinquefolium (American ginseng).).).).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.28 no.4
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• pp.497-503
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• 1983

This study was conducted to define the effects of light intensity on the amount of saponin and free sugar and the ratio of triol group saponin and diol group saponin (PT/PD) in the leaf of Panax ginseng C.A. Meyer. 4-Year-old ginseng plants were grown under the shadings of different light transmittance rate(LTR) of 5%, 10%, 20% and 30% for 5 months and the leafiets were sampled from 2nd low at late August to determine the amount of saponin and free sugar.-Rd was main ginseuoside in the diol group saponin but in triol group saponin, ginsenoside-Re showed highest value and next was ginsenoside-$Rg_1$ and $Rg_2$ respectively. Up to 20% of light transmittance rate (LTR), the ginseng leaves grown under high light intensity showed an increase in the amount of total saponin and the ratio of PT/PD but the amount of total saponin and the ratio of PT/PD but the amount of total saponin and the ratio of PT/PD was decreased at the ginseng leaves grown under the shading of 30% LTR. The ginseng leaves grown under the shading of 20% LTR showed a significant increase in the amount of glucose and fructose but a significant decrease of sucrose content. A significant positive correlation ($r=0.992$^{**}$) was recognized between the of amount of total saponin and glucose.

• Journal of Ginseng Research
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• v.43 no.4
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• pp.562-571
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• 2019

Background: Ginsenosides have been widely used clinically for many years and were regarded as very safe. However, a few researches on the toxicities of these kinds of agents showed that some ginsenosides may have side-effect on the rats or dogs. So it is extremely necessary to further clarify the potential toxicity of ginsenosides. This study was carried out to investigate long-term toxicity and genotoxicity of 25-methoxydammarane-3, 12, 20-triol ($25-OCH_3-PPD$), a new derivative of ginsenoside, in beagle dogs. Methods: Twenty-four beagle dogs were divided randomly into four treatment groups and repeatedly orally administered with $25-OCH_3-PPD$ capsule at 60, 120, and 240 mg/kg/day for 91 consecutive days. Ames, micronucleus, and chromosomal aberration tests were established to analyze the possible genotoxicity of $25-OCH_3-PPD$. Results: There was no $25-OCH_3-PPD$einduced systemic toxicity in beagle dogs at any doses. The level of $25-OCH_3-PPD$ at which no adverse effects were observed was found to be 240 mg/kg/day. The result of Ames test showed that there was no significant increase in the number of revertant colonies of $25-OCH_3-PPD$ administrated groups compared to the vehicle control group. There were also no significant differences between $25-OCH_3-PPD$ administrated groups at all dose levels and negative group in the micronucleus test and chromosomal aberration assay. Conclusion: The highest dose level of $25-OCH_3-PPD$ at which no adverse effects were observed was found to be 240 mg/kg per day, and it is not a genotoxic agent either in somatic cells or germs cells. $25-OCH_3-PPD$ is an extremely safe candidate compound for antitumor treatment.

• Journal of Ginseng Research
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• v.15 no.3
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• pp.257-262
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• 1991

1) 인삼 사포닌을 5% 황산으로 가수분해하여 TMS화 한 후 GC로 분석한 결과 인삼중의 모든 사포닌을 diol계 사포닌과 triol계 사포닌으로 나누어 분석할 수 있었다. 2) 시료중의 전체 diol계(PD)와 triol계(PT) 사포닌을 ginsenoside Rb$_1$과 Rg$_1$의 양으로 각각 환산하여 표시하고 미삼에는 백삼에 비해 diol계 사포닌의 함량이 많은 것을 이용하여 PD/PT글 구하고 여기에서 구해진 비를 이용하여 시료중의 백삼 및 미삼의 이론적 함량을 구할 수 있었다. 3) 이 방법의 검출 한계는 백삼의 양으로 0.14$\mu$g이었다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.11 no.3
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• pp.81-86
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• 1982

Ginseng saponins separated from Panax ginseng C.A. Meyer were analyzed by high-performance liquid chromatography using a carbohydrate analysis column. The effect on the resolution and retention time of each ginsenosides, as well as the addition effect of n-butanol on a acetonitrile/water system, was examined using various proportions of acetonitrile/water system (80/20-94/6) which have been used with typical solvent of carbohydrate analysis column. The retention time of each ginsenosides was greatly affected by the compositions and mixture proportions of the mobile phase and also markedly increased as the proprotion of acetonitrile in mobile phase increased. It was proved that acetonitrile/water system (80/20) and acetonitrile/water/n-butanol system (86/14/10) were very effective mobile phases for diol and triol sapoin analyses, respectively. According to the result obtained by this method, the PT/PD ratios of white and red ginseng saponins were 0.401 % and 0.561 %, respectively. Red ginseng increased PT/PD ratio, compared with white ginseng. This is attributed to the change of saponin pattern by processing method and the change of PT/PD ratio would greatly influence on the biochemical and pharmacological effects of ginseng and its products.

• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.181-188
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• 1978

To study on the changes in saponins from callus mass by tissue culture, the callus was derived from the petiole of Korean Ginseng (Panax Ginseng C.A. Meyer) and cultivated on Murashige and Skoog's agar medium supplemented with 2.4-dichlorophenoxyacetic acid and kinetin for 8 months. Then, well-grown callus was analyzed for its components estimation. The results obtained are as follows: (1) When saponins isolated from callus mass were chromatographed on a silca gel plate, and determined by the thinchrograph TFG-10, the ratio of Rb, c to Rg(f) in saponins was 2.16 to 1 and Rb, c, d to Re, g (f) was 1 to 1.63, while in the case of saponins from the root of Panax Ginseng grown by soil culture, the ratio of Rb, c to Rg(f) was 1.03 to 1 and the ratio of Rb, c,d to Re, g(f) was 1 to 1.17. (2) Sapogenins were obtained from the hydrolysates of saponins, and determined by thinchrograph TFG-10. The ratio of panaxadiol to panaxatriol in sapogenins from callus saponins was 2.66 to 1, while the ratio of panaxadiol to panaxatriol in sapogenins from ginseng root saponins was 1.86 to 1. From the results above mentioned, we concluded that the relative contents of sapogenins in saponins from callus mass by tissue culture were different from those in saponins from ginseng root by soil culture.

• Proceedings of the Ginseng society Conference
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• 1980.09a
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• pp.151-170
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• 1980

Physiological response of Panax ginseng var. atropurpureacaulo (purple stem variety, Pg) to light was reviewed through old literatures and recent experiments. Canopy structure, growth, pigment, leaf anatomy, disease occurence, transpiration, photosynthesis (PS), leaf saponin, photoperiodism and nutrient uptake were concerned. P. ginseng var. xanthocarpus (yellow berry variety, Px) and Panax quinquefolius(Pq) were compared with Pg if possible. Compensation point(Cp) increased with increase of light and ranged from 110 to 150 at $20^{\circ}C$ but from 140 to 220 at $30^{\circ}C$ with 4 to 15 Klux indicating occurence of light and temperature-dependent high photorespiration. Characteristics of Korea ginseng to hate high temperature was well accordance with an observation 2000 years ago. Korea ginseng showed lower Cp and appeared to be more tolerant to high light intensity and temperature than American sheng although the latter showed greater PS, stomata frequency and conductance, chlorophyll and carotenoids. Px showed lower PS than Pg probably due to higher Cp. Total leaf saponin was higher in leaves grown under high light. Ratio or diol saponin and triol saponin(PT/PD) decreased with increase of light intensity during growing mainly due to decrease of ginsenoside $Rg_1$ but increase of ginsenoside Rd. Leaves of Pg and Px had $Rg_1$ but no $Rb_3$ which was only found as much as $20\%$ of total in Pq leaves, and decreased with increase of light intensity. Re increased in Pg and Px but decreased in Pq with increase of light. PT/PD in leaf ranged 1.0-1.5 in Pg and Px but around 0.5 in Pq. Korea ginseng has Yang characteristics(tolerant to high light and temperature), cultured under Eum(shade) condition and long been used for Yang efficacy (to build up energy) while Pq was quite contrary. Traditional low light $intensity(3-8\%)$ for Korea ginseng culture appeared to be strongly related to historical unique quality. Effect of light quality and photoperiodism was not well known. Experiences are long but scientific knowledge is short for production and quality assessment of ginseng. Recent scientific knowledge of ginseng should learn wisdom from old experiences.

• Proceedings of the Ginseng society Conference
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• 2007.05a
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• pp.23-24
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• 2007

Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$를 $20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.

• Journal of Ginseng Research
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• v.42 no.2
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• pp.133-143
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• 2018

Background: AD-2 (20(R)-dammarane-3b, 12b, 20, 25-tetrol; 25-OH-PPD) is a ginsenoside and isolated from Panax ginseng, showing anticancer activity against extensive human cancer cell lines. In this study, effects and mechanisms of 1C ((20R)-3b-O-(L-alanyl)-dammarane-12b, 20, 25-triol), a modified version of AD-2, were evaluated for its development as a novel anticancer drug. Methods: MTT assay was performed to evaluate cell cytotoxic activity. Cell cycle and levels of reactive oxygen species (ROS) were determined using flow cytometry analysis. Western blotting was employed to analyze signaling pathways. Results: 1C concentration-dependently reduces prostate cancer cell viability without affecting normal human gastric epithelial cell line-1 viability. In LNCaP prostate cancer cells, 1C triggered apoptosis via Bcl-2 family-mediated mitochondria pathway, downregulated expression of mouse double minute 2, upregulated expression of p53 and stimulated ROS production. ROS scavenger, N-acetylcysteine, can attenuate 1C-induced apoptosis. 1C also inhibited the proliferation of LNCaP cells through inhibition on $Wnt/{\beta}-catenin$ signaling pathway. Conclusion: 1C shows obvious anticancer activity based on inducing cell apoptosis by Bcl-2 family-mediated mitochondria pathway and ROS production, inhibiting $Wnt/{\beta}-catenin$ signaling pathway. These findings demonstrate that 1C may provide leads as a potential agent for cancer therapy.