Analytical Optimum of Ginsenosides according to the Gradient Elution of Mobile Phase in High Performance Liquid Chromatography

HPLC의 이동상 용매조건에 따른 인삼 Ginsenoside 분석

  • 박지영 (부산대학교 생명자원과학부) ;
  • 원준연 (중부대학교 교양학과) ;
  • 이충열 (부산대학교 생명자원과학부)
  • Published : 2007.06.30

Abstract

This study was conducted to analyze not only for the quality guaranteed of red ginseng but also for the minor ginsenosides. Although several studies have reported to analyze ginseng saponins, those were focused to major saponins, including 6 to 7 ginsenosides. As increase of interest in medicinal effect of ginseng products, anasis of various ginsenosides in both red and white ginseng are strongly demanded. To perform optital condition of 12 ginsenoside analysis, We controlled HPLC conditions, such as the gradient elution of the mobile phase. We found the adequate separation method for 12 ginse-nosides. The optimum condition was as following : H$_2$O/CH$_3$CN ratios were 82/18, 70/30, 55/45 and 50/50, respectively. Sol-vent flow rate was 1.00 ma/min. Column temperature was kept to 35$^{\circ}$C. UV detector was set to 203 nm.

HPLC에 의한 미량 Ginsenoside의 분석법을 확립하고자 연구를 수행하였던 바, 그 결과를 요약하면 다음과 같다. 인삼 ginsenosid의 분석은 많이 시도되어 왔으나 주요 사포닌 및 미량 사포닌의 분석법이 정립되지 않고 있는 실정인바, 본 연구에서는 이동상 용매의 기울기 용리를 다양하게 변화를 주어 Rb$_1$, Rb$_2$, Rb$_3$, Rc, Rd, Re, Rf Rg$_1$, Rg$_2$, Rg$_3$, Rh$_1$, Rh$_2$의 총 12종의 사포닌을 양호하게 분리하는데 성공하였다. 이동상의 흐름 속도는 1.00ml/min이고 column온도는 35$^{\circ}$C, UV detector의 파장은 203nm로 모두 일정하게 한 결과였으며, 최적의 분석조건은 H$_2$O와 CH$_3$CN의 용매 조성이 82/18, 70/30, 55/45, 50/50으로 각각의 ginsenosides의 안정적인 Area 값을 얻을 수 있었고 재현성을 시도해본 결과, 반복간 편차가 적어 재현성이 매우 양호한 것으로 나타났다.

Keywords

References

  1. An YN, Lee SY, Choung MG, Kang KH (2002a) Optimum Harvesting time based on growth characteristics of four-year ginseng. Korean J. Crop Sci. 47(3):211-215
  2. An YN, Lee SY, Choung MG, Choi KJ, Kang KH (2002b) ginsenoside concentration and chemical component as affected by harvestin time of four-year ginseng root. Korean J. Med. Crop Sci. 47(3):216-220
  3. Attele, S, Wu J, Yuan CS (1999) Ginseng phasrmacology : multiple constituents and multiple actions. Biochem. Pharmacol. 58: 1685-1693 https://doi.org/10.1016/S0006-2952(99)00212-9
  4. Jang JG, Lee KS, Kwon DW, Nam KY, Choi JH (1983) Study on the changes of saponin contents in relation to root age of Panax ginseng. Korean J. Food & Nurition 12(1):37-40
  5. Jeong CM, Bae KK (1999) Effect of $CO_2$ enrichment on the differentiation of multi-shoots and saponin contents in tissue culture of korea gunsen. Korean J. Med. Crop Sci. 7(4):296-302
  6. Jeong SI, Kim CS, Lee NW, Lee YG, Lee HS, Kim IK (1998) Separation Of 20 (R&S) prosaponins isomers of ginsenoside-Rg2 and Rg3 from ginseng saponins by reversed-phase high performance liquid chromatography. Analytical Science & Technology vol. 11(5):404-408
  7. Kim CS, Kim SB (2001) Determination of ginseng saponins by reveesed-phase high performance liquid chromatography. Korean J. Med. Crop Sci. 9(1):21-25
  8. Kim MW, Ko SR, Choi KJ, Kim SC (1987) Distribution of saponin in various section of Panax ginseng root and changes of its contents according to root age. Korean J. Ginseng Sci. 11(1):10-16
  9. Lee SE, Lee SW, Bang JK, Yu YJ, Seong NS (2004) Antioxidant activities of leaf, stem and root of panax ginseng C. A. Meyer. Korean J. Med. Crop Sci. 12(3):237-242
  10. Lee SW, Kang SW, Kim DY, Seong NS, Park HW (2004) comparison of growth characteristics and compounds of ginseng cultivated by paddy and upland cultivation. Korean J. Med. Crop Sci. 12(1):10-16
  11. Lee SW, Cha SW, Hyun DY, Kim YC, Kang SW, Seung NS (2005) Comparison of growth characteristics, and extract and crude saponin contents in 4-Year-Old ginseng cultured by direct seeding and transplanting cultivation. Korean J. Med. Crop Sci. 13(6):241-244
  12. Park MK, Park JH, Kang JS, Lee MY, Park YI, Yu SJ, Han BH (1993) Rapid hydrolysis of ginseng saponin by microwave oven reaction. Kor. J. Ginseng Sci., 17(1):35-38
  13. Shibata S, Tanaka O, Soma K, lita Y, Ando T, Nakamura H (1965) Studies on saponins and sapogenins of ginseng: the structure of panaxatriol. Tetrahedron Lett 3:207-213
  14. Shibata S, Tanaka O (1966) Protopanaxadiol a genuine sapogenins of ginseng saponins. Chem Pharm Bull 14:595-600 https://doi.org/10.1248/cpb.14.595
  15. 고려삼의 이해 (1995) 고려인삼학회 p. 9-16
  16. 고려인삼 (1994) 한국인삼연초연구원, p. 63-88