Method Development for the Profiling Analysis of Endogenous Metabolites by Accurate-Mass Quadrupole Time-of-Flight(Q-TOF) LC/MS

LC/TOFMS를 이용한 생체시료의 내인성 대사체 분석법 개발

  • Lee, In-Sun (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Kim, Jin-Ho (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Cho, Soo-Yeul (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Shim, Sun-Bo (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Park, Hye-Jin (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Lee, Jin-Hee (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Lee, Ji-Hyun (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Hwang, In-Sun (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Kim, Sung-Il (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Lee, Jung-Hee (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Cho, Su-Yeon (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Choi, Don-Woong (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team) ;
  • Cho, Yang-Ha (Korea Food & Drug Administration, National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Department, Advanced Analysis Team)
  • 이인선 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 김진호 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 조수열 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 심선보 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 박혜진 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 이진희 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 이지현 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 황인선 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 김성일 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 이정희 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 조수연 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 최돈웅 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀) ;
  • 조양하 (식품의약품안전청 식품의약품안전평가원 독성평가연구부 첨단분석팀)
  • Received : 2010.07.06
  • Accepted : 2010.11.30
  • Published : 2010.12.31

Abstract

Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.

Keywords

References

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