Analytical Science and Technology (분석과학)

The Korean Society of Analytical Sciences (한국분석과학회)
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Validation of LC-MS/MS method for determination of ginsenoside Rg1 in human plasma

인체 혈장 중 Ginsenoside Rg1의 정량을 위한 LC-MS/MS 분석법 검증

  • Kim, Yunjeong (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Han, Song-Hee (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Jeon, Ji-Young (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Hwang, Min-Ho (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Im, Yong-Jin (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Lee, Sun Young (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Chae, Soo-Wan (Biomedical Research Institute of Chonbuk National University Hospital) ;
  • Kim, Min-Gul (Biomedical Research Institute of Chonbuk National University Hospital)
  • 김윤정 (전북대학교병원 의생명연구원) ;
  • 한송희 (전북대학교병원 의생명연구원) ;
  • 전지영 (전북대학교병원 의생명연구원) ;
  • 황민호 (전북대학교병원 의생명연구원) ;
  • 임용진 (전북대학교병원 의생명연구원) ;
  • 이선영 (전북대학교병원 의생명연구원) ;
  • 채수완 (전북대학교병원 의생명연구원) ;
  • 김민걸 (전북대학교병원 의생명연구원)
  • Received : 2013.03.06
  • Accepted : 2013.06.10
  • Published : 2013.08.25
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Abstract

A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the investigation of the ginsenoside Rg1 in human plasma. After addition of internal standard (digoxin), plasma was diluted with acetone and methanol (80:20), the supernatant was concentrated and analyzed by LC-MS/MS. The optimal chromatographic separation was achieved on an Agilent Eclipse XDB-C18 column ($4.6{\times}150mm$, $5{\mu}m$) with a mobile phase of 0.1% formic acid in water and 0.1% formic acid in methanol at a flow rate of 0.9 mL/min gradient mode. The standard calibration curve for ginsenoside Rg1 was linear ($r^2=0.9995$) over the concentration range 1~500 ng/mL in human plasma. The intra- and inter-day precision over the concentration range of ginsenoside Rg1 was lower than 7.53% (correlation of variance, CV), and accuracy exceeded 98.28%. This LC-MS/MS assay of ginsenoside Rg1 in human plasma is applicable for quantifying in the pharmacokinetic study.

LC-MS/MS를 이용하여 인체 혈장 중 ginsenoside Rg1의 신속하고 정확한 분석법을 개발하고 이 분석법에 대한 검증을 수행하였다. 혈장 분석을 위하여 internal standard인 digoxin을 첨가한 후 acetone과 methanol용액 (80:20)으로 전처리하고, 그 상층액을 진공농축 한 후, LC-MS/MS로 분석하였다. 최적 크로마토그래피 분석은 0.1% formic acid 첨가된 water와 methanol을 이동상으로 하여 Agilent Eclipse XDB-C18 column ($4.6{\times}150mm$, $5{\mu}m$)을 이용하여 0.9 mL/min의 유속으로 gradient mode로 수행하였다. 혈장 중 ginsenoside Rg1의 표준 검량선은 1~500 ng/mL의 농도 범위에서 우수한 직선성($r^2=0.9995$)을 보였으며, 일내, 일간 정밀성은 변동계수 7.53% 이하, 정확성은 98.28% 이상이었다. 결과적으로 본 분석법은 ginsenoside Rg1의 약동학 연구에 적용되기에 충분한 감도와 특이성, 직선성, 정밀성 및 정확성을 가지고 있음을 확인하였다.

Keywords

References

  1. J. T. Baker, R. P. Borris, B. Carte, G. A. Cordell, D. D. Soejarto, G. M. Cragg, M. P. Gupta, M. M. Iwu, D. R. Madulid and V. E. Tyler, J. Nat. Prod., 58(9), 1325-1357 (1995). https://doi.org/10.1021/np50123a003
  2. C. S. Yuan, C. Z. Wang, S. M. Wicks and L. W. Qi, J. Ginseng Res., 34(3), 160-167 (2010). https://doi.org/10.5142/jgr.2010.34.3.160
  3. M. Mochizuki, Y. C. Yoo, K. Matsuzawa, K. Sato, I. Saiki, S. Tono-oka, K. Samukawa and I. Azuma, Biol. Pharm. Bull., 18(9), 1197-1202 (1995). https://doi.org/10.1248/bpb.18.1197
  4. T. Yokozawa, T. Kobayashi, H. Oura, and Y. Kawashima, Chem. Pharm. Bull., 33(2), 869-872 (1985). https://doi.org/10.1248/cpb.33.869
  5. K. Takagi, H. Saito and H. Nabata, Jpn. J. Pharmacol., 22(2), 245-249 (1972). https://doi.org/10.1254/jjp.22.245
  6. P. M. Stavro, M. Woo, T. F. Heim, L. A. Leiter and V. Vuksan, Hypertension, 46(2), 406-411 (2005). https://doi.org/10.1161/01.HYP.0000173424.77483.1e
  7. C. Pan, Y. Huo, X. An, G. Singh, M. Chen, Z. Yang, J. Pu and J. Li, Vascul. Pharmacol., 56(3-4), 150-158 (2012). https://doi.org/10.1016/j.vph.2011.12.006
  8. H. Saito, Y. Yoshida,and K. Takagi, Jpn. J. Pharmacol., 24(1), 119-127 (1974). https://doi.org/10.1254/jjp.24.119
  9. B. X. Wang, J. C. Cui, A. J. Liu and S. K. Wu, J. Tradit. Chin. Med., 3(2), 89-94 (1983).
  10. H. Matsuda, K. Samukawa and M. Kubo, Planta Med., 56(1), 19-23 (1990). https://doi.org/10.1055/s-2006-960875
  11. P. Hu, G. A. Luo, Q. Wang, Z. Z. Zhao, W. Wang and Z. H. Jiang, Arch. Pharm. Res., 31(10), 1265-1273 (2008). https://doi.org/10.1007/s12272-001-2105-2
  12. '건강기능식품의 기준 및 규격 전면 개정(식품의약품안정청고시 제 2008-12호)', 식품의약품안전청.
  13. '인삼 및 인삼제품-진세노사이드(Rb1, Rg1)함량 측정-고속액체크로마토그래프법(KS H 2153)', 산업자원부 기술표준원.
  14. H. Oura, S. Hiai, Y. Odaka and T. Yokozawa, J. Biochem., 77(5), 1057-1065 (1975). https://doi.org/10.1093/oxfordjournals.jbchem.a130806
  15. O. Sticher and F. Soldati, Planta Med., 36(1), 30-42 (1979). https://doi.org/10.1055/s-0028-1097237
  16. T. T. Yip, C. N. Lau, P. P. But and Y. C. Kong, Am. J. Chin. Med., 13(1-4), 77-88 (1985). https://doi.org/10.1142/S0192415X85000125
  17. N. Fuzzati, J. Chromatogr. B., 812(1-2), 119-133 (2004). https://doi.org/10.1016/j.jchromb.2004.07.039
  18. S. I. Jeong, C. S. Kim, Y. G. Lee, H. S. Lee and I. K. Kim, Anal. Sci. Technol., 11(5), 404-407 (1998).
  19. S. I. Jeong, C. S. Kim, N. W. Lee, K. J. Choi, Y. G. Lee and I. K. Kim, Anal. Sci. Technol. 11(6), 436-439 (1998).
  20. W. Chen, Y. Dang and C. Zhu, Chin. Med., 5, 12 (2010). https://doi.org/10.1186/1749-8546-5-12
  21. J. Sun, G. Wang, X. Haitang, L. Hao, P. Guoyu and I. Tucker, J. pharmaceut. Biomed., 38(1), 126-132 (2005). https://doi.org/10.1016/j.jpba.2004.12.007
  22. M. Song, S. Zhang, X. Xu, T. Hang and L. Jia, J. Chromatogr. B., 878(32), 3331-3337 (2010). https://doi.org/10.1016/j.jchromb.2010.10.007
  23. L. Liu, J.Huang, X. Hu, K. Li and C. Sun J. Chromatogr. B., 879(22), 2011-2017 (2011). https://doi.org/10.1016/j.jchromb.2011.05.018
  24. J. Zhao, C. Su, C. Yang, M. Liu, L. Tang, W. Su, Z. and Liu J. Pharm. Biomed. Anal., 64-65, 94-97 (2012). https://doi.org/10.1016/j.jpba.2012.02.017
  25. D. N. Patel, H. S. Lin and H. L. Koh J. Mass Spectrum., 47, 1510-1517 (2012). https://doi.org/10.1002/jms.3095