This facility provides services for analysis of metabolites and lipids.
We perform quantitative and semi-quantitative analysis of known metabolites using LC-MS/MS(MRM) system Triple Quadrupole with an ion trap (QTRAP 6500+ from Sciex). We can also perform quantification of metabolites using stable-isotope labeled internal standards or semi-quantitative analysis using external standards upon request.
We have developed protocols to monitor over 300 metabolites in a targeted manner. For high-sensitivity analysis, we can perform the analysis of the specific metabolites using optimized protocols. The different metabolites can also have different sample requirements.
We perform global profiling of lipids and metabolites in an untargeted manner which can be used for broad profiling or for discovery studies (video).
Metabolomics
Untargeted metabolomics is performed using the Orbitrap Exploris 240 high-resolution mass spectrometer (ThermoFisher) in positive and/or negative ionization mode coupled with ultra-high performance liquid chromatography (Vanquish UHPLC) in HILIC separation mode.
HILIC (ESI+/-ve) (please see a case study published in Cell Stem Cell)
Lipidomics
Lipidomics and free fatty acids analysis are performed using the Orbitrap Exploris 240 high-resolution mass spectrometer (ThermoFisher) coupled with ultra-high performance liquid chromatography (Vanquish UHPLC) in reversed-phase (RP) separation mode.
We perform structural identification of metabolic features of interest by matching acquired data against our in-house EMBL-MCF spectral library, comprising ~300 metabolites, which allows us to provide identification with highest confidence (Level I ID – via accurate mass, isotopic pattern, MS/MS spectra, retention time). In addition, the analysis of unknown features of interest can be supplemented by utilizing other public libraries to annotate compounds (Level II ID – without retention time).
For lipid identification we utilize a public library (MS-DIAL LipidBlast) covering over 100 lipid classes and over 1.3 million spectra. In addition, we evaluate elution patterns of homologous lipid species within lipid classes to ensure highest level confidence in identification.
We perform the detection and quantitation of drug and drug metabolites from various biological samples for pharmacokinetics and drug metabolism analysis (non-GLP), biotransformation, and bioaccumulation screenings. For this, we use standard calibration curves and internal standards based quantitative analysis using high-resolution Q-Exactive Plus (ThermoFisher) mass spectrometer coupled with ultra-high performance liquid chromatography (Vanquish UHPLC) in reversed-phase (RP) and/or HILIC separation mode.
When there are no methods available or existing methods fail to achieve the required sensitivity, coverage or reliability, we can perform method development tailored for specific applications e.g. analysis of drugs and drug metabolites, modified metabolites, plant samples as well as special classes of metabolites at an additional cost.