Glycerophospholipid Identification and Quantitation by Electrospray Ionization Mass Spectrometry
ABSTRACT Glycerophospholipids are the structural building blocks of the cellular membrane. In addition to creating a protective barrier around the cell, lipids are precursors of intracellular signaling molecules that modulate membrane trafficking and are involved in transmembrane signal transduction. Phospholipids are also increasingly recognized as important participants in the regulation and control of cellular function and disease. Analysis and characterization of lipid species by mass spectrometry (MS) have evolved and advanced with improvements in instrumentation and technology. Key advances, including the development of "soft" ionization techniques for MS such as electrospray ionization (ESI), matrix-assisted laser desorption/ionization (MALDI), and tandem mass spectrometry (MS/MS), have facilitated the analysis of complex lipid mixtures by overcoming the earlier limitations. ESI-MS has become the technique of choice for the analysis of multi-component mixtures of lipids from biological samples due to its exceptional sensitivity and capacity for high throughput. This chapter covers qualitative and quantitative MS methods used for the elucidation of glycerophospholipid identity and quantity in cell or tissue extracts. Sections are included on the extraction, MS analysis, and data analysis of glycerophospholipids and polyphosphoinositides.
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- "This approach allowed for the quantification of both of the PL classes and the lipid species (fatty acid composition) within a PL class. Ivanova et al. described a negative mode LC–ESI-MS linear ion trap method for the analysis of PLs in cell cultures and tissues . These authors applied ion trap scans and PL class selective internal standards for the quantification of PLs. "
ABSTRACT: A new, sensitive and selective liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS/MS) method was developed for the analysis of Phospholipids (PLs) in bio-oils and fats. This analysis employs hydrophilic interaction liquid chromatography-scheduled multiple reaction monitoring (HILIC-sMRM) with a ZIC-cHILIC column. Eight PL class selective internal standards (homologs) were used for the semi-quantification of 14 PL classes for the first time. More than 400 scheduled MRMs were used for the measurement of PLs with a run time of 34min. The method's performance was evaluated for vegetable oil, animal fat and algae oil. The averaged within-run precision and between-run precision were ≤10% for all of the PL classes that had a direct homologue as an internal standard. The method accuracy was generally within 80-120% for the tested PL analytes in all three sample matrices. Copyright © 2015 Elsevier B.V. All rights reserved.Journal of Chromatography B 08/2015; 1001:140-149. DOI:10.1016/j.jchromb.2015.07.036 · 2.69 Impact Factor
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- "GP, also known as phospholipids, are ubiquitous in nature and are important components of lipid bilayers and are involved with cell signaling and metabolism. Based on the nature of the polar head group at the sn-3 position of the glycerol backbone in eukaryotes and eubacteria, or the sn-1 position in the case of archaebacteria , GP may be subdivided into distinct classes including glycerophosphocholines, glycerophosphatidic acids, glycerophosphoglycerols (PG), glycerophosphoserines (PS), PE, and PI. Brain tissue contains relatively high G, and alterations of their composition have been implicated in neurologic disorders . "
ABSTRACT: Due to the incidence of type-2 diabetes and hypertension, chronic kidney disease (CKD) has emerged as a major public health problem worldwide. CKD results in premature death from accelerated cardiovascular disease and various other complications. Early detection, careful monitoring of renal function, and response to therapeutic intervention are critical for prevention of CKD progression and its complications. Unfortunately, traditional biomarkers of renal function are insufficiently sensitive or specific to detect early stages of disease when therapeutic intervention is most effective. Therefore, more sensitive biomarkers of kidney disease are needed for early diagnosis, monitoring, and effective treatment. CKD results in profound changes in lipid and lipoprotein metabolism that, in turn, contribute to progression of CKD and its cardiovascular complications. Lipids and lipid-derived metabolites play diverse and critically important roles in the structure and function of cells, tissues, and biofluids. Lipidomics is a branch of metabolomics, which encompasses the global study of lipids and their biologic function in health and disease including identification of biomarkers for diagnosis, prognosis, prevention, and therapeutic response for various diseases. This review summarizes recent developments in lipidomics and its application to various kidney diseases including chronic glomerulonephritis, IgA nephropathy, chronic renal failure, renal cell carcinoma, diabetic nephropathy, and acute renal failure in clinical and experimental research. Analytical technologies, data analysis, as well as currently known metabolic biomarkers of kidney diseases are addressed. Future perspectives and potential limitations of lipidomics are discussed. © 2015 Elsevier Inc. All rights reserved.Advances in clinical chemistry 02/2015; 68:153-175. DOI:10.1016/bs.acc.2014.11.002 · 4.30 Impact Factor
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- "This article is protected by copyright. All rights reserved HILIC-LC-MS was performed with an internal standard to confirm the ion variations observed in the MS spectra according to the Lipid Maps methods (Ivanova et al., 2007). Amounts of internal standard were added according to the percentage of each PL class in the total PL extract: "
ABSTRACT: Altered phospholipid (PL) metabolism has been associated with pregnancy disorders. Moreover, lipid molecules such as endocannabinoids (eCBs) and prostaglandins (PGs) are important mediators of reproductive events. In humans, abnormal decidualization has been linked with unexplained infertility, miscarriage and endometrial pathologies. Anandamide (AEA), the major eCB, induces apoptosis in rat decidual cells. In this study, the PL profile of rat decidual cells was characterized by a Mass spectrometry (MS) based lipidomic approach. Furthermore, we analyzed a possible correlation between changes in PL of rat decidual cells' membrane and AEA-induced apoptosis. We found an increase in phosphatidylserine and a reduction of cardiolipin and phophatidylinositol relative contents. In addition, we observed an increase in the content of alkyl(alkenyl)acylPL, plasmalogens, and of long chain fatty acids especially with high degrees of unsaturation, as well as an increase in lipid hydroperoxides in treated cells. These findings provide novel insights on deregulation of lipid metabolism by anandamide, which may display further implications in decidualization process. This article is protected by copyright. All rights reservedJournal of Cellular Physiology 12/2014; 230(7). DOI:10.1002/jcp.24901 · 3.87 Impact Factor