• Title/Summary/Keyword: Analysis of ginsenoside

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Optimization of ginseng hairy roots culture and its ginsenoside analysis

  • Ji, Joong Gu;Yoo, Sun Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.995-1002
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    • 2018
  • Hairy root culture of ginseng is industrially prospected because the cultivation period of ginseng is relatively long. In this study, the effect of medium concentration and sucrose concentration on hairy root culture of ginseng was evaluated. The optimization of ginseng hairy roots transformed by Agrobacterium rhizogene were performed liquid medium. The MS(Murashinge & Skoog basal medium) concentration was selected with 1/2 strength MS and the optimal sucrose concentration was determined at 2-3%(w/v). At the optimum culture condition, The yield (the ratio of weight of grown hairy root cultures to weight of fresh ginseng hairy roots) and production rate of ginseng root were 19.42 times and 5.73 g/l-day. The major ginsenosides were Rb group, Re and Rg1. The produced total ginsenoside content in the solid medium was 9.87 (mg/g) and increased 1.34 times in the liquid medium (13.23 mg/g). In solid culture, the contents of ginsenosides Rb, Re and Rg1 were 2.14, 3.65 and 1.87 mg/g, respectively. In liquid culture, the contents of ginsenosides Rb, Re and Rg1 were 3.54, 4.12 and 2.63 mg/g, respectively.

Antiviral effects of Korean Red Ginseng on human coronavirus OC43

  • Chi Hwan Jeong;Jisu Kim;Bo Kyeong Kim;Kang Bin Dan;Hyeyoung Min
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.329-336
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    • 2023
  • Background: Panax ginseng Meyer is a medicinal plant well-known for its antiviral activities against various viruses, but its antiviral effect on coronavirus has not yet been studied thoroughly. The antiviral activity of Korean Red Ginseng (KRG) and ten ginsenosides against Human coronavirus OC43 (HCoV-OC43) was investigated in vitro. Methods: The antiviral response and mechanism of action of KRG extract and ginsenoside Rc, Re, Rf, Rg1, Rg2-20 (R) and -20 (S), Rg3-20 (R) and -20 (S), and Rh2-20 (R) and -20 (S), against the human coronavirus strain OC43 were investigated by using plaque assay, time of addition assay, real-time PCR, and FACS analysis. Results: Virus plaque formation was reduced in KRG extract-treated and HCoV-OC43-infected HCT-8 cells. KRG extract decreased the viral proteins (Nucleocapsid protein and Spike protein) and mRNA (N and M gene) expression, while increased the expression of interferon genes. Conclusion: KRG extract exhibits antiviral activity by enhancing the expression of interferons and can be used in treating infections caused by HCoV-OC43.

HPLC SEPARATION AND QUANTITATIVE DETERMINATION OF GINSENOSIDES FROM PANAX GINSENG, PANAX QUINQUEFOLIUM AND FROM GINSENG DRUG PREPARATIONS

  • Soldati F
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.59-69
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    • 1980
  • A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

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Correlation analysis between growth characteristics and ginsenoside contents of 4-year-old wild-simulated ginseng (Panax ginseng C.A. Meyer) with different cultivation sites (지역별 4년근 산양삼의 생육특성 및 진세노사이드 함량 간의 상관관계 분석)

  • Yeong-Bae Yun;Jeong-Hoon Huh;Dae-Hui Jeong;Jiah Kim;Yurry Um
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.253-259
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    • 2022
  • The aim of this study was to investigate the correlation between growth characteristics and ginsenoside contents of 4-year-old wild-simulated ginseng cultivated in different regions. Most of the soil properties except for available phosphate showed significantly higher in Pyeongchang than in other cultivation sites. The growth characteristics except for root length and number of rootlets showed significantly higher in Pyeongchang than in other cultivation sites. In the case of 8 ginsenoside contents, the content of F2-AS was significantly higher in Muju than in other cultivation sites and the content of F1 in Yeongju was significantly high. In Yeongwol, the contents of Rb1 and Re-p were significantly high and the content of Ro in Pyeongchang showed significantly higher than in other cultivation sites. Root length and soil pH did not show a significant correlation with any soil properties and growth characteristics of wild-simulated ginseng, respectively. Most of the growth characteristics showed significantly positive correlations with electrical conductivity, organic matter content, total nitrogen content, exchangeable cations (K+, Ca2+, Mg2+), and cation exchange capacity. Rb1 and Re-p showed significantly negative correlations with most of the growth characteristics of wild-simulated ginseng except for the number of rootlets. Ro showed a significantly positive correlation with stem length, number of leaflets per stem, leaflet length, leaflet width, and root diameter. The results of this study probably will help to provide useful information on the establish a quality standard by investigate correlation analysis between growth characteristics and ginsenoside content of 4-year-old wild-simulated ginseng.

Analysis of the Marker Compounds in Sagunja-tang by LC-ESI-MS (LC-ESI-MS에 의한 사군자탕의 지표성분 분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.50 no.1
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    • pp.65-71
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    • 2019
  • One of the oriental medicine prescriptions, Sagunja-tang consists of four herbal medicines (Ginseng Radix, Poria Sclerotium, Atractylodis Rhizoma Alba, and Glycyrrhiziae Radix et Rhizoma) and has been used as a medicine to enhance tonify the function of spleen and stomach in Korea. In this study, we conducted simultaneous analysis of the 9 marker components, liquiritin apioside, liquiritin, ginsenoside Rg1, liquiritigenin, ginsenoside Rb1, glycyrrhizin, atractylenolide III, atractylenolide II, and atractylenolide I in Sagunja-tang using a liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Marker compounds were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, 1.7 mm) and the column was maintained at $45^{\circ}C$. The mobile phase consists of 0.1% (v/v) aqueous formic acid and acetonitrile with gradient condition. The LC-MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS system with multiple reaction monitoring (MRM) method in the positive and negative modes. The calibration curves of the nine marker components showed good linearity with coefficient of determination ${\geq}0.9984$ within tested range. The limits of detection and limits of quantification values were 0.27-2.42 ng/mL and 0.81-7.27 ng/mL, respectively. The concentrations of tested 9 analytes in the lyophilized Sagunja-tang sample using the established LC-ESI-MS/MS MRM method were detected up to 16.593 mg/g. These results can be useful as a basic data for the quality control of an oriental medicine prescriptions.

Quantification Analysis and Antioxidant Activity of Leejung-tang (LC-MS/MS를 이용한 이중탕(理中湯)의 정량분석 및 항산화 활성)

  • Seo, Chang-Seob;Kim, Ohn Soon;Kim, Yeji;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.21 no.1
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    • pp.177-185
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    • 2013
  • Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.

Effect of Ginseng Saponin on LDL Receptor Biosynthesis (인삼사포닌의 저밀도지질단백질(LDL)수용체에 미치는 영향)

  • Joo Chung No;Lee Hee Bong;Lee Yong Woo;Kang In Chul
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.47-54
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    • 1988
  • Cholesterol a component of all eucaryotic plasma membranes. is essential for the growth and viability of cells in higher organisms. However. too much cholesterol can be lethal because of atherosclerosis resulting from the deposition of cholesterol ester plaques. It was attempted in this study to understand the preventive effect of ginseng saponin. one of the major components of the roots of Panax ginseng C.A. Meyer. against hypercholesterolemia induced by high cholesterol diet. $^{125}I-LDL$ was injected intravenously to rabbits and rats. which were fed a high cholesterol diet with and/or without ginseng saponin for 12 days. The disappearance of the radioactivity occurred faster in the test group than the control. The effect of saponin fraction from Panax ginseng C.A. Meyer and the purified ginsenosilks. $Rb_1,\;Rb_2,\;Re\;and\;Rg_1,$ on LDL receptor biosynthesis in high cholesterol fed rat has been investigated. Analysis of LDL receptors from various organs such as liver. kidney. adrenal cortex and testis showed that the population of LDL receptors of test group significantly higher than that of the control. It was also found that liver homogenate containing ginsenosides $(10^{-3}-10^{-4}\%)$ stimulated the biosynthesis of bile acid form cholesterol. From the above results. it seemed that ginsenosides lower the cholesterol level by stimulating cholesterol metabolism. which result in the suppression of the inhibitory action of cholesterol on LDL receptor biosynthesis.

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Quality and Functional Properties of Red Ginseng Prepared with Different Steaming Time and Drying Methods (원료삼의 증삼 및 건조 조건별 홍삼의 품질 및 기능성)

  • Kim, Kyo-Youn;Shin, Jin-Ki;Lee, Su-Won;Yoon, Sung-Ran;Chung, Hun-Sik;Jeong, Yong-Jin;Choi, Myung-Sook;Lee, Chi-Moo;Moon, Kwang-Deog;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.39 no.5
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    • pp.494-499
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    • 2007
  • The quality and functional properties of red ginseng in relation to steaming and drying conditions were evaluated. Fresh ginseng (5-year roots), cultivated in the Punggi region, were steamed for 2.5, 3.5, or 4.5 hr, and then dried by hot-air (60-$65^{\circ}C$/24 hr and $40^{\circ}C$,/3-4d) freezing ($-80^{\circ}C$/56 hr), and infrared (900 W/$62^{\circ}C$/68 hr). Hunter#s yellowness (b-value) and browning indexes (420 nm) of the samples were higher in the rootlets than in the main roots. Furthermore, these same index values were found to be high in the order of 3.5, 4.5, and 2.5 hr and infrared, hot-air, and freezing for steaming and subsequent drying, respectively. Analysis of soluble solids, total phenolics, total flavonoids, acidic polysaccharides, and electron donating abilities of the steamed and dried samples showed that 3.5hr of steaming with infrared drying was optimal. However, crude saponin contents were not influenced by steaming and drying conditions. The contents of $ginsenoside-Rg_l$, -Re, -Rf and $-Rb_2$, which were the major components in the samples, were reduced with steaming time, while the amounts of $-Rg_3$ and $-Rh_2$ increased, reaching the highest levels at 3.5 and 4.5 hr in the main roots and rootlets, respectively. The contents of $-Rg_3$ and $-Rh_2$ were similar in both the freeze-dried and hot-air dried samples.

A systematic exploration of ginsenoside Rg5 reveals anti-inflammatory functions in airway mucosa cells

  • Hyojin Heo;Yumin Kim;Byungsun Cha;Sofia Brito;Haneul Kim;Hyunjin Kim;Bassiratou M. Fatombi;So Young Jung;So Min Lee;Lei Lei;Sang Hun Lee;Geon-woo Park;Byeong-Mun Kwak;Bum-Ho Bin;Ji-Hwan Park;Mi-Gi Lee
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.97-105
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    • 2023
  • Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.

Validation on the Analytical Method of Ginsenosides in Red Ginseng

  • Cho B. G.;Nho K. B.;Shon H. J.;Choi K. J.;Lee S. K.;Kim S. C;Ko S. R.;Xie P. S.;Yan Y. Z.;Yang J. W.
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.491-501
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    • 2002
  • A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.

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