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

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

  • 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)
  • 한송희 (전북대학교병원 의생명연구원) ;
  • 김윤정 (전북대학교병원 의생명연구원) ;
  • 전지영 (전북대학교병원 의생명연구원) ;
  • 황민호 (전북대학교병원 의생명연구원) ;
  • 임용진 (전북대학교병원 의생명연구원) ;
  • 이선영 (전북대학교병원 의생명연구원) ;
  • 채수완 (전북대학교병원 의생명연구원) ;
  • 김민걸 (전북대학교병원 의생명연구원)
  • Received : 2012.08.22
  • Accepted : 2012.11.13
  • Published : 2012.12.31

Abstract

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.

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.

Keywords

References

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