Subject Area物理化学
Metabolic profiling of plasma by gas chromatography/time-of-flight mass spectrometry: method develop and validation
Li X(李响); Lu X(路鑫); Xu GW(许国旺)
Conference Namethe 13th Beijing Conference and Exhibition on Instrumental Analysis
Conference Date2009-11-25
Conference Place中国
Funding Organization中国分析测试协会
AbstractD59 METABOLIC PROFILING OF PLASMA BY GAS CHROMATOGRAPHY/TIME-OF-FLIGHT MASS SPECTROMETRY: METHOD DEVELOP AND VALIDATION Xiang LI, Xin LU, Guowang XU National Chromatographic R & A Center, Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, Dalian 116023, China Metabonomics has been developed to investigate the metabolic changes coupled with pathophysiologic stimuli or genetic modification. Gas chromatography/time-of-flight mass spectrometry (GC-TOFMS) has the advantages of high sensitivity, large linear range and fast acquisition rates, which are suitable for comprehensive, non-target analysis of biofluids in metabonomics. In our study, a metabolic profiling method of plasma is developed and validated with GC-TOFMS, indicating high sensitivity, good resolution and high throughput. 1 EXPERIMENTAL Three internal standards were added into plasma. Methanol was added in, and then the mixture was centrifuged to discard protein. Supernatant was lyophilized and derivatized by bis(trimethylsilyl)trifluoroacetamide (BSTFA, pyridine was added as solvent). Plasma samples were analyzed by a LECO Pegasus 4D GC×GC-TOFMS instrument (LECO Corporation, used in GC-TOFMS mode). A 30m×250 μm×0.25 μm DB-5MS column was used in the Agilent 6890N GC. Helium was used as the carrier gas. Mass spectra of m/z 33–600 were collected. 2 RESULTS Sample preparation and chromatographic conditions were optimized based on the peak number, peak intensity and the resolution of both internal standards and plasma metabolites. An optimal result of separation and detection of the metabolites could be gained with the method above (Figure 1). Three internal standards were well distributed in the chromatogram. The metabolites could be separated with high resolution, including more than two hundred metabolites. To validate the method, limit of detection, recovery, linearity and precision were calculated according to the three internal standards. A good recovery (79.83±8.16%), precision (<5%) and linear range (R2>0.98) could be gained. These results indicated that a comprehensive and accurate metabolic profiling could be achieved by our method. This method could be further applied to wide fields of metabonomics researches as a standard operating procedure. Figure 1. Typical GC-TOFMS chromatogram of plasma. REFERENCES 1. Philippe A. Guy, Isabelle Tavazzi, Stephen J. Bruce, Ziad Ramadan, Sunil Kochhar. J. Chromatogr. B, 871: 253–260 (2008). 2. Paul Begley, Sue Francis-McIntyre, Warwick B. Dunn, David I. Broadhurst, Antony Halsall, Andy Tseng, Joshua Knowles, HUSERMET Consortium, Royston Goodacre, Douglas B. Kell. Anal. Chem. 81: 7038–7046 (2009) 3. Joachim Kopka. J. Biotechnol. 124: 312–322 (2006).
Document Type会议论文
Corresponding AuthorXu GW(许国旺)
Recommended Citation
GB/T 7714
Li X,Lu X,Xu GW. Metabolic profiling of plasma by gas chromatography/time-of-flight mass spectrometry: method develop and validation[C],2009:466/2.
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