Equipment

Liquid chromatography hyphenated to high resolution /high accuracy mass spectrometry for untargeted lipidomics:

Orbitrap Elite coupled with a Shimadzu Nexera (UPLC): Lipid classes can be chromatographically separated by normal phase LC, based upon head group polarity, or by reverse phase regarding the lipids acyl chain length and number of unsaturations.  This orthogonal approach also relies on the mass accuracy of the mass spectrometer (< 10 ppm), which corresponds to predict the exact elemental composition of the lipids up to the third decimal place.

Velos

Liquid chromatography or gas chromatography hyphenated to triple quadrupole mass spectrometers for targeted lipidomics:

AB SCIEX 6500 QTRAP coupled with Shimadzu Prominence HPLC is used for multiple reaction monitoring (MRM) assays. This system is more sensitive than the orbitrap Elite and Velos Pro allowing semi and absolute quantitation of lipid subclasses in the range of  pg in sample.

The SCIEX 6500 can be coupled with a differential ion mobility SelexION device. Mobility separation provides an extra level of selectivity between the chromatography and the mass spectrometer for a further degree of separation of coeluting compounds, isomers, and low abundant compounds buried in the background noise.

Ab Sciex 6500

Shotgun throughput lipidomics analysis:

Lipid extracts, diluted in a mass spectrometry compatible solvent, are directly infused into the Velos Pro using nanoelectrospray ionisation (nESI) by the NanoMate robot. This approach can be used for high throughput analysis of the major/abundant lipid subclasses for large number samples. LESA (Liquid Extraction Surface Analysis) of tissues or structural characterisation of small analytes/metabolites/lipids, and peptides can also be performed using nESI.

NanoMate

Nano-liquid chromatography hyphenated to Velos Pro mass spectrometer for selective reaction monitoring (SRM):

This instrumentation setup provides chromatographic separation analytes/metabolites and peptides when sample amount is very low.  This system will be available for open access by BI institute staff interested on performing their own analysis. Training on calibration, operation, and cleaning will be provided prior to booking the instrument. Training on protein digestion/sample preparation, chromatography set-up, and data analysis can be also be facilitated by lipidomics staff to support the proteomics facility internal demand.

Please contact us if you are interested: lipidomics@babraham.ac.uk

Orbitrap

Gas chromatography hyphenated to TSQ Quantum mass spectrometer for multiple reaction monitoring (MRM):

Instrumentation used for the analysis of small volatile analytes, forexample derivatised Fatty acid methyl esters (FAMEs).  The mass spectrometer has an electron ionisation source, which allow structure elucidation of the analytes. These instrument is shared with the biological chemistry facility.

TS Quantum