Metabolite identification spectrometry

Multiple mass analyzers exist that can perform tandem mass spectrometry. Some use a tandem-in-space configuration, such as the triple quadrupole mass analyzers illustrated (Fig.3.9). Others use a tandem-in-time configuration and include instruments such as ion-traps (ITMS) and Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS or FTMS). A triple quadrupole mass spectrometer can only perform the tandem process once for an isolated precursor ion (e.g., MS/MS), but trapping or tandem-in-time instruments can perform repetitive tandem mass spectrometry (MS ), thus adding n 1 degrees of structural characterization and elucidation. When an ion-trap is combined with HPLC and photodiode array detection, the net result is a profiling tool that is a powerful tool for both metabolite profiling and metabolite identification. 

Dear G. Plumb R. Mallet D. Use of monolithic silica columns to increase analytical throughput for metabolite identification by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry, 2001, 15, 152-158. 

Clarke, N., Cox, K., Dunn-Meynell, K., Korfmacher, W., Lin, C. C., White, R. and Cayen, M., Strategies for Semi-Automated Metabolite Identification in Drug Discovery Using LC-MS", American Society for Mass Spectrometry 1999 Conference Abstract, Dallas, TX, USA, 1999. 

Rindgen, D., Cox, K., Clarke, N. and Korfmacher, W., An Integrated Approach to Metabolite Identification for the Drug Discovery Compound SCH 123 using the Triple Quadrupole, Ion Trap and Q-TOF Instruments, American Society for Mass Spectrometry 2000 Conference Abstract, Long Beach, CA, USA, 2000. 

Hopfgartner, G. Zell, M. Q trap MS a new tool for metabolite identification, in Using Mass Spectrometry for Drug Metabolism Studies, ed. Korf-macher, W. A., CRC Press, 2004. 

Pelander, A. Ojanpera, 1. Laks, S. Rasanen, 1. Vuori, E. Toxicological Screening with Formula-Based Metabolite Identification by Liquid Chromatography/Time-of-Flight Mass Spectrometry. Anal. Chem. 2003, 75, 5710-5718. 

The presence of 5 chlorines on the aromatic ring simplified metabolite identification by mass spectrometry. 

Ma, S., Chowdhury, S.K. and Alton, K.B. (2006) Application of Mass Spectrometry for Metabolite Identification. Current Drug Metabolism, 7, 503-523. 

The marriage of HPLC to mass spectrometry (MS), now developed into a mature instrumentation, continues to greatly impact many of the separation sciences, especially in pharmaceutical analysis where it has been used in new drug discovery [23,24] and in drug metabolite identification [25-27]. HPLC-MS has also made an impact on lipid research, providing a convenient approach to the analysis of phospholipids and fatty acids [28,29]. It has also greatly benefited the field of proteomics [30-34], especially analysis of protein structure and function. 

Ma SG, Chowdhury SK, Alton KB. Application of mass spectrometry for metabolite identification. Current Drug Metabolism 7, 503-523, 2006. 

The system relies upon preliminary fractionation of the microbial crude extract by dualmode countercurrent chromatography coupled with photodiode array detection (PDA). The ultraviolet-visible (UV-Vis) spectra and liquid chromatography-mass spectrometry (LC-MS) of biologically active peaks are used for identification. Confirmation of compound identity is accomplished by nuclear magnetic resonance (NMR). Use of an integrated system countercurrent chromatography (CCC) separation, PDA detection, and LC-MS rapidly provided profiles and structural information extremely useful for metabolite identification (dereplication, Figure 14.1). 

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Hop, C. E., Tiller, P. R., and Romanyshyn, L. (2002). In vitro metabolite identification using fast gradient high performance liquid chromatography combined with tandem mass spectrometry. Rapid Commun. Mass Spectrom. 16 212-219. 

Kantharaj, E., Ehmer, P. B., Tuytelaars, A., Van Vlaslaer, A., Mackie, C., and Gilissen, R. A. (2005b). Simultaneous measurement of metabolic stability and metabolite identification of 7-methoxymethylthiazolo[3,2-a]pyrimidin-5-one derivatives in human liver microsomes using liquid chromatography/ion-trap mass spectrometry. Rapid Commun. Mass Spectrom. 19 1069-1074. 

Tiller, P. R., Yu, S., Castro-Perez, J., Fillgrove, K. L., and Baillie, T. A. (2008). High-throughput, accurate mass liquid chromatography/tandem mass spectrometry on a quadrupole time-of-flight system as a first-line approach for metabolite identification studies. Rapid Commun. Mass Spectrom. 22 1053-1061. 

Liu, D. Q., Xia, Y. Q., and Bakhtiar, R. (2002). Use of a liquid chromatography/ion trap mass spectrometry/triple quadrupole mass spectrometry system for metabolite identification. Rapid Commun. Mass Spectrom. 16 1330-1336. 

Rourick, R. A., Jenkins, K. M., Walsh, J. P., Xu, R., Cai, Z., and Kassel, D. B. (2002). Integration of custom LC/MS automated data processing strategies for the rapid assessment of metabolic stability and metabolite identification in drug discovery. In Proceedings of the 50th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL. 

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Figure 2 The two most frequently used approaches for metabolomics are NMR spectroscopy and mass spectrometry. To maximize the coverage of the metabolome, these techniques can be combined. This figure shows the analysis of the aqueous fraction of a section of heart tissue that uses NMR spectroscopy, gas chromatography mass spectrometry and liquid chromatography mass spectrometry. Although mass spectrometry approaches are inherently more sensitive than NMR spectroscopy, metabolite identification becomes more difficult.

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