• Journal of Ginseng Research
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• 제43권1호
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• pp.105-115
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• 2019

Background: Ginsenosides with less sugar moieties may exhibit the better adsorptive capacity and more pharmacological activities. Methods: An efficient method for the separation of four minor saponins, including gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, from Panax notoginseng leaves (PNL) was established using biotransformation, macroporous resins, and subsequent preparative high-performance liquid chromatography. Results: The dried PNL powder was immersed in the distilled water at $50^{\circ}C$ for 30 min for converting the major saponins, ginsenosides Rb1, Rc, Rb2, and Rb3, to minor saponins, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, respectively, by the enzymes present in PNL. The adsorption characteristics of these minor saponins on five types of macroporous resins, D-101, DA-201, DM-301, X-5, and S-8, were evaluated and compared. Among them, D-101 was selected due to the best adsorption and desorption properties. Under the optimized conditions, the fraction containing the four target saponins was separated by D-101 resin. Subsequently, the target minor saponins were individually separated and purified by preparative high-performance liquid chromatography with a reversed-phase column. Conclusion: Our study provides a simple and efficient method for the preparation of these four minor saponins from PNL, which will be potential for industrial applications.

• Journal of Ginseng Research
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• 제42권2호
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1988년도 학술대회지
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• pp.141-146
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• 1988

북미가 원산지인 왜생삼(Panax trifolius L.)은 인삼속(오가과)에 속하며 카나다 남부에서 북미에 걸쳐 서식한다. 왜생삼 잎에서 4종류의 프라보느이드와 5종류의 진세노사이드로 확인된 총 9종류의 화합물을 분리하였다. 2종류의 진세노사이드로 확인된 총 9종류의 화합물을 분리하였다. 2종류의 프라보노이드는 kaempferol-3,7-dirhamnoside와 kaempferol-3-gluco-7rhamnoside로 각각 확인되었다. 4종류의 진세노사이드는 각각 notoginsenoside-Fe, ginsenoside-Rd, ginsenoside-Rc와 ginsenoside-$Rb_3$로 확인되었으며, 이들 왜생삼의 ginsenoside 공통된 골격구조는 (20S)-protopanaxadiol인 것으로 밝혀졌다. 프라보노이드와 진세노사드의 동정은 왜생삼의 뿌리, 줄기, 잎, 꽃과 열매에서 추출하여 2차원 박층 그로마토그라피(2D-TLC)와 고압 액체크로마토그라피(HPLC)로 하였다. 왜생삼과 근연종인 고려인삼(Panax ginseng C.A. Meyer)및 미국삼(Panx quinquefolium L.)의 뿌리, 줄기, 잎, 꽃과 열매에서 추출한 프라보노이드와 진세노시드의 정량은 고압 액체크로마토그래피 만을 사용하여 분석하였다. 화합물 1,3과 4로 명명한 kaempferol 유도체인 프라보노이드 3가지는 왜생삼의 뿌리에 $10.8\%,\;2.8\%$와 $8.4\%$가 각기 함유되어 있었으나 고려인삼과 미국삼에서는 함유되어 있지 않았다. 오가과 인삼속식물 뿌리에서 프라보노이드가 확인, 동정된 것은 이것이 처음이다.

• Journal of Ginseng Research
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• 제40권4호
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• pp.344-350
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• 2016

Background: Mountain-cultivated ginseng (MCG) and cultivated ginseng (CG) both belong to Panax ginseng and have similar ingredients. However, their pharmacological activities are different due to their significantly different growth environments. Methods: An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS)-based approach was developed to distinguish MCG and CG. Multivariate statistical methods, such as principal component analysis and supervised orthogonal partial-least-squares discrimination analysis were used to select the influential components. Results: Under optimized UPLC-QTOF-MS/MS conditions, 40 ginsenosides in both MCG and CG were unambiguously identified and tentatively assigned. The results showed that the characteristic components of CG and MCG included ginsenoside Ra3/isomer, gypenoside XVII, quinquenoside R1, ginsenoside Ra7, notoginsenoside Fe, ginsenoside Ra2, ginsenoside Rs6/Rs7, malonyl ginsenoside Rc, malonyl ginsenoside Rb1, malonyl ginsenoside Rb2, palmitoleic acid, and ethyl linoleate. The malony ginsenosides are abundant in CG, but higher levels of the minor ginsenosides were detected in MCG. Conclusion: This is the first time that the differences between CG and MCG have been observed systematically at the chemical level. Our results suggested that using the identified characteristic components as chemical markers to identify different ginseng products is effective and viable.

• Journal of Applied Biological Chemistry
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• 제61권4호
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• pp.371-377
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• 2018

인삼(Panax. ginseng) 열매를 80% MeOH 수용액으로 3회 반복 추출한 뒤, 감압 농축한 추출물을 EtOAc, n-BuOH과 $H_2O$ 층으로 계통 분획을 실시하였다. EtOAc분획에 대하여 $SiO_2$ 및 ODS column chromatography를 반복실시하여 5종의 ginsenoside 화합물을 분리 및 정제하였다. NMR, IR, FAB/MS 데이터를 해석하여, 각각 ginsenoside F1 (1), ginsenoside F2 (2), ginsenpside F3 (3), ginsenoside Ia (4) 및 notoginsenoside Fe (5)로 구조 동정 하였다. 화합물 2-5는 인삼열매에서는 이번에 처음 분리 보고되었다. 분리한 5종의 화합물을 인체 암세포주(HCT-116, SK-OV-3, HeLa, HepG2, SK-MEL-5)에 처리하여 세포독성을 측정하였다. 이 중 화합물 2, 4, 및 5가 인체 암세포주에 대해 세포독성을 저해시키는 것을 알 수 있었다. 화합물 2는 SK-MEL-5, HepG2, HeLa세포에서 $IC_{50}$값이 82.8, 86.8, $78.3{\mu}M$로 확인되었다. 화합물 4는 HCT-116, SK-MEL-5, SK-OV-3, HepG2, HeLa 세포에서 $IC_{50}$ 값이 24.5, 25.4, 26.3, 22.0, $24.9{\mu}M$로 확인되었다. 화합물 5는 SK-MEL-5 세포에서 $IC_{50}$ 값이 $81.7{\mu}M$로 확인되었다. 인삼 열매에서 분리한 화합물2, 4, 및 5가 암세포주에 대해 강한 세포독성을 나타내는 것을 확인하였으며, 이 화합물들은 공통적으로 3번 수산기에 glucopyranose를 가지고 있음을 확인하였다.