Novel Application of Reversed-Phase UPLC-oaTOF-MS for Lipid Analysis in Complex Biological Mixtures:  A New Tool for Lipidomics

Abstract

Ultra-Performance LC (UPLC) utilizing sub-2-mum porous stationary phase particles operating with high linear velocities at pressures >9000 psi was coupled with orthogonal acceleration time-of-flight (oaTOF) mass spectrometry and successfully employed for the rapid separation of lipids from complex matrices. The UPLC system produced information-rich chromatograms with typical measured peak widths of 3 s at peak base, generating peak capacities in excess of 200 in 10 min. Further UPLC coupled with MSE technology provided parent and fragment mass information of lipids in one chromatographic run, thus, providing an attractive alternative to current LC methods for targeted lipid analysis as well as lipidomic studies.

A shotgun lipidomics approach in Sinorhizobium meliloti as a tool in functional genomics

“…Most methods for lipid analysis employ HPLC separation schemes (33,(52)(53)(54)(55), and when the separation of lipid classes and their molecular components is desired, a combination of NP-HPLC followed by RP-HPLC is needed (12). The research presented here allowed the analysis of eight lipid classes in crude lipid extracts of S. meliloti using a shotgun lipidomics approach.…”

UPLC-based metabonomic applications for discovering biomarkers of diseases in clinical chemistry

“…In this way, two mass chromatograms are generated, one with information on the intact molecules from the first function, and the other with the fragmented ion information from the second function. A variety of data-processing algorithms can be used to extract metabolite information from these data [12,13]. In other words, MS E can provide parallel alternating scans for acquisition at either low collision energies to obtain precursor ion information or high collision energies to obtain full-scan accurate mass fragment, precursor ion, and small neutral molecules.…”

An Emerging Role for Metabolomics in Nutrition Science

“…Usually, liquid chromatography tools are used to separate and screen complex mixtures of metabolites extracted from biological samples. In this example, metabolites were separated using UPLC coupled with a hybrid QToF system mass spectrometer [80,81]. The analysis provided a metabolite profile, which is a biochemical snapshot of the metabolite inventory of the tissue under investigation.…”

“…Next, the components are separated using chromatographic techniques such as liquid chromatography and gas chromatography or directly infused in a mass spectrometry (MS) detector [80,81]. Mass spectrometers that offer high resolution and high accuracy are generally used, including the time of flight, Fourier transform ion cyclotron resonance, and orbitrap.…”