DOI QR코드

DOI QR Code

LC-MS/MS를 이용한 인체 혈장에서 Ginsenoside Rb1의 분석법 검증

Validation of the LC-MS/MS Method for Ginsenoside Rb1 Analysis in Human Plasma

  • 한송희 (전북대학교병원 의생명연구원) ;
  • 김윤정 (전북대학교병원 의생명연구원) ;
  • 전지영 (전북대학교병원 의생명연구원) ;
  • 황민호 (전북대학교병원 의생명연구원) ;
  • 임용진 (전북대학교병원 의생명연구원) ;
  • 이선영 (전북대학교병원 의생명연구원) ;
  • 채수완 (전북대학교병원 의생명연구원) ;
  • 김민걸 (전북대학교병원 의생명연구원)
  • Han, Song-Hee (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Kim, Yunjeong (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Jeon, Ji-Young (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Hwang, Minho (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Im, Yong-Jin (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Lee, Sun Young (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Chae, Soo-Wan (Biomedical Research Institute, Chonbuk National University Hospital) ;
  • Kim, Min-Gul (Biomedical Research Institute, Chonbuk National University Hospital)
  • 투고 : 2012.08.22
  • 심사 : 2012.11.13
  • 발행 : 2012.12.31

초록

LC-MS/MS를 사용함으로써 인체 혈장에서 ginsenoside Rb1의 분석법을 개발하고 검증하였다. 유속 0.9 mL/min에 이동상 0.1% formic acid가 첨가된 water와 methanol을 사용하여 기울기 용리 조건으로 설정하였으며 사용한 분석 칼럼은 C 18($4.6mm{\times}150mm$, particle size 5 ${\mu}m$)을 사용하여 분리하였다. MRM(multiple reaction monitoring) 방법의 전기 분무 이온화 이온 분석기로 모니터링 하여 분석하였다. 인체 혈장 샘플은 액체-액체 추출방법에 의해 acetone과 water가 섞인 용액으로 추출하였다. 이 분석의 검량선 범위는 10~500 ng/mL이며 상관계수는 0.9995를 나타냈다. 일내, 일간의 정밀성 농도범위는 상관계수 5.8% 그리고 정확성은 96.0~104.6%로 나타났다. 이 LC-MS/MS를 이용한 인체 혈장의 ginsenoside Rb1의 연구가 약동학 연구에 적용할 수 있을 거라 생각한다.

A new liquid chromatographic tandem mass spectrometric (LC-MS/MS) assay for the quantification of ginsenoside Rb1 in human plasma was developed and validated. The separation was performed on a Agilent C18 column ($4.6mm{\times}150mm$, particle size 5 ${\mu}m$) with a gradient elution of 0.1% formic acid in water and 0.1% formic acid in methanol and a flow rate of 0.9 mL/min. The analyte was determined using electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode (m/z 1131.714${\rightarrow}$365.303). Human plasma samples were extracted with acetone : water (50:50) by the liquid-liquid extraction method. The method was linear over the dynamic range of 10~500 ng/mL with a correlation coefficient of r=0.9995. The intra-and inter-day precision over the concentration range of ginsenoside Rb1 was lower than 5.8% (correlation of variance, CV), and the accuracy was between 96.0~104.6%. This LC-MS/MS assay of ginsenoside Rb1 in human plasma is applicable for quantification in a pharmacokinetic study.

키워드

참고문헌

  1. Wen J, Zimmer EA. 1996. Phylogeny and biogeography of Panax L. (the ginseng genus, araliaceae): inferences from ITS sequences of nuclear ribosomal DNA. Mol Phylogenet Evol 6: 167-177. https://doi.org/10.1006/mpev.1996.0069
  2. Li XG. 1992. Studies on the transforming mechanism of amino acid components in ginseng in the course of ginseng process. Korean J Ginseng Sci 16: 64-67.
  3. Singh VK, Agarwal SS, Gupta BM. 1984. Immunomodulatory activity of Panax ginseng extract. Planta Med 50: 462-465. https://doi.org/10.1055/s-2007-969773
  4. Benishin CG. 1992. Actions of ginsenoside Rb1 on choline uptake in central cholinergic nerve endings. Neurochem Int 21: 1-5. https://doi.org/10.1016/0197-0186(92)90061-U
  5. Kaneko H, Nakanishi K. 2004. Proof of the mysterious efficacy of ginseng: basic and clinical trials: clinical effects of medical ginseng, Korean red ginseng: specifically, its anti-stress action for prevention of disease. J Pharmacol Sci 95: 158-162. https://doi.org/10.1254/jphs.FMJ04001X5
  6. Kim WY, Kim JM, Han SB, Lee SK, Kim ND, Park MK, Kim CK, Park JH. 2000. Steaming of ginseng at high temperature enhances biological activity. J Nat Prod 63: 1702-1704. https://doi.org/10.1021/np990152b
  7. Kim H, Chen X, Gillis CN. 1992. Ginsenosides protect pulmonary vascular endothelium against free radical-induced injury. Biochem Biophys Res Commun 189: 670-676. https://doi.org/10.1016/0006-291X(92)92253-T
  8. Hyun MS, Hur JM, Shin YS, Song BJ, Mun YJ, Woo WH. 2009. Comparison study of white ginseng, red ginseng, and fermented red ginseng on the protective effect of LPS-induced inflammation in RAW 264.7 cells. J Appl Biol Chem 52: 21-27. https://doi.org/10.3839/jabc.2009.004
  9. Park JY, Lee CY, Won JY. 2007. Analytical optimum of ginsenosides according to the gradient elution of mobile phase in high performance liquid chromatography. Korean J Medicinal Crop Sci 15: 215-219.
  10. Choi EH, Lee HJ, Kim CJ, Kim JT, Kwun IS, Kim Y. 2004. Anti-stress effects of ginseng in immobilization-stressed rats. J Food Sci Nutr 9: 253-258. https://doi.org/10.3746/jfn.2004.9.3.253
  11. Choi GJ. 1991. Components of raw ginseng and quality control. Korean J Ginseng Sci 12: 247-256.
  12. Han YJ, Kwon KR, Cha BC, Kwon OM. 2007. Component analysis of cultivated ginseng, cultivated wild ginseng, and natural wild ginseng by structural parts using HPLC method. Korean Pharmacopuncture 10: 37-53.