Direct LC-MS/MS Analysis of Extra- and Intracellular Glycerophosphoinositol in Model Cancer Cell Lines

Abstract

Glycerophosphoinositols (GPIs) are water-soluble bioactive phospholipid derivatives of increasing interest as intracellular and paracrine mediators of eukaryotic cell functions. The most representative compound of the family is glycerophosphoinositol (GroPIns), an ubiquitous component of mammalian cells that participates in cell proliferation, cell survival and cell response to stimuli. Levels and activity of this compound vary among cell types and deciphering these functions requires accurate measurements in in vitro and in vivo models. The conventional approaches for the analysis of GroPIns pose several issues in terms of sensitivity and product resolution, especially when the product is in the extracellular milieu. Here we present an UPLC-MS study for the quantitative analysis of this lipid derivative in cells and, for the first time, culture supernatants. The method is based on a solid-phase extraction that allows for fast desalting and analyte concentration. The robustness of the procedure was tested on the simultaneous measurements of intra- and extracellular levels of GroPIns in a number of human cell lines where it has been shown that the non-transformed cells are characterized by high extracellular level of GroPIns, whereas the tumor cells tended to have higher intracellular levels.

Study on plasma metabolomics profiling of depression in Chinese community-dwelling older adults based on untargeted LC/GC‒MS

“…Our results revealed that 1-(sn-glycero-3-phospho)-1D-myo-inositol had a high diagnostic value. 1-(sn-Glycero-3-phospho)-1D-myo-inositol, also known as glycerophosphoinositol, is produced through membrane phosphatidylinositol through two successive deacylation steps catalysed by phospholipase A2IVα 26 , 27 . According to a study, high glycerophosphoinositol levels indicate cellular phenomena associated with the activation of RAS/mitogen-activated protein kinase (MAPK) pathways 27 .…”

Raman spectroscopy and mass spectrometry identifies a unique group of epidermal lipids in active discoid lupus erythematosus

“…The current mismatch of these m/z may be a reflection of the mechanistic differences in ionsation between MALDI and electrospray ionisation. Furthermore, it has been reported that triacylglycerides are difficult to detect with MALDI, while the water soluble glycerophosphoinositols poses problems for conventional chromatographic protocols in terms of resolution and reproducibility, and these species may not be triacylglycerides 28 , 32 . Further work could elucidate these putative lipid identities.…”

Dysregulation of lipid metabolism in the pseudolobule promotes region-specific autophagy in hepatitis B liver cirrhosis

“…The lysoPC mainly comes from the circulation produced by PC via phospholipase A2. 20 , 21 Since the expression level of PC was significantly reduced, the byproduct of lysoPC would also decline. While comparing within the subregions of the samples, the expression of PC and choline was found to be significantly downregulated in the PL group (Figure 7 A).…”