Equipment

Q Exactive Plus benchtop Orbitrap mass spectrometer

Q Exactive Plus Orbitrap Mass Spectrometer, equipped with an Ion Max source and a HESI II probe (Thermo Scientific).

 

This system is capable of measuring mass-to-charge ratios with less than 5 ppm error and a resolution of up to 140,000 and can also collect MS/MS fragmentation data.

Mass spectrometer is coupled to a Dionex UltiMate 3000 ultra-high performance liquid chromatography (UPLC) system designed for pressures up to 1000 bar, permitting rapid separation times of ~30 minutes per sample.

Together with a library of several hundred synthetic standards, the combination of exact mass, fragmentation and chromatographic retention time data permits the positive identification of most common cellular metabolites.

Operating the mass spectrometers in either targeted or full-scan mode enables both hypothesis-driven and hypothesis-generating (untargeted) experiments to be performed.

What can be analyzed:

  • Routine measurements of many polar metabolites (including amino acids, glycolytic and TCA cycle intermediates) can be performed (based on the inclusion list)

  • Novel methods for the detection of the metabolites OFF-the list could be developed. The sample preparation, chromatography and/or mass spectrometry portions of the analysis may be optimized or newly developed.

 

Several levels of analysis are available:

  • Relative differences in metabolite concentrations between groups of samples are sufficient for many applications

  • Absolute (molar) quantitation is performed using standard curves

  • Untargeted analysis, which is performed using Sieve 2.2 software (Thermo) and XCMS