Validation of an LC–MS/MS method for the quantification of choline-related compounds and phospholipids in foods and tissues

Xiong, Yeping; Zhao, Yuanyuan; Goruk, Sue; Oilund, Kirsten; Field, Catherine J.; Jacobs, René L.; Curtis, Jonathan M.

doi:10.1016/j.jchromb.2012.10.038

Cited by 80 publications

(45 citation statements)

References 28 publications

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“…LCeMS/MS provides useful structural information for the analytes, even in the presence of background contaminants found in complex biological systems. Several PCOOH metabolites from human plasma and tissues were previously examined and identified using LCeMS/MS with MRM [13]. Here we applied this analytical method to analyze oxidatively modified PCs such as PCOOH, PCOH, PONPC, PAzPC and others that might be present in the cells or the culture medium.…”

Section: Discussionmentioning

confidence: 99%

Metabolism and cytotoxic effects of phosphatidylcholine hydroperoxide in human hepatoma HepG2 cells

Suzuki

Nakagawa

Kato

et al. 2015

Biochemical and Biophysical Research Communications

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Section: Discussionmentioning

confidence: 99%

Metabolism and cytotoxic effects of phosphatidylcholine hydroperoxide in human hepatoma HepG2 cells

Suzuki

Nakagawa

Kato

et al. 2015

Biochemical and Biophysical Research Communications

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“…v/v). The choline metabolites were analyzed directly by LC-MS [35], while the ethanolamine and serine metabolites were first converted to dansylated derivatives [36] which were then quantified with LC-MS using SRM [34].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

“…Canada),35 S-methionine from Perkin Elmer, phospholipase D (Streptomyces sp. ), formic acid, ammonium formate and ammonium hydroxide from Sigma.…”

mentioning

confidence: 99%

The PNPLA-family phospholipases involved in glycerophospholipid homeostasis of HeLa cells

Hermansson

Hänninen

Hokynar

et al. 2016

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Mammalian cells maintain the glycerophospholipid (GPL) compositions of their membranes nearly constant. To achieve this, GPL synthesis and degradation must be coordinated. There is strong evidence that A-type phospholipases (PLAs) are key players in homeostatic degradation of GPLs, but the identities of the PLAs involved have not been established. However, some members of the Patatin-like phospholipase domain-containing proteins (PNPLAs) have been implicated. Accordingly, we knocked down all the PNPLAs significantly expressed in human HeLa cells using RNA interference and then determined whether the turnover of the major glycerophospholipids is affected by using mass spectrometry and metabolic labeling with stable isotope-labeled precursors. Knockdown of PNPLA9, PNPLA6 or PNPLA4 significantly (30-50%) reduced the turnover of phosphatidylcholine, -ethanolamine and -serine. In a notable contrast, turnover of phosphatidylinositol was not significantly affected by the knockdown of any PNPLA. Depletion of PNPLA9 and PNPLA4 also inhibited G0/G1 to S cell cycle progression, which could thus be regulated by GPL turnover. These results strongly suggest that PNPLA9, -6 and -4 play a key role in GPL turnover and homeostasis in human cells. A hypothetical model suggesting how these enzymes could recognize the relative concentration of the different GPLs is proposed.

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“…The adopted analytical approach is based on high-resolution FTMS, performed on the lipid extracts of farmed gilthead sea bream (Sparus aurata) after a hydrophilic interaction liquid chromatography (HILIC) separation achieved on a polar stationary phase (silica). As recently demonstrated for the phospholipidome of human platelets and blood microparticles [25,26], HILIC is able to provide a class separation of PLs, whose elution order is dictated by the nature of the polar head [27][28][29][30][31][32]. LPL originating from the two main PL classes, phosphatidylcholines (PC) and phosphatidylethanolamines (PE), are wholly separated both from their precursors and other PL classes, thus avoiding possible interference with isobaric species of different PL classes.…”

Section: Introductionmentioning

confidence: 99%

Identification of isobaric lyso-phosphatidylcholines in lipid extracts of gilthead sea bream (Sparus aurata) fillets by hydrophilic interaction liquid chromatography coupled to high-resolution Fourier-transform mass spectrometry

et al. 2015

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The numerous and varied biological roles of phosphatidylcholines (PC) and lysophosphatidylcholines (LPC) have fueled a great demand for technologies that enable rapid, in-depth structural examination of these lipids in foodstuffs. Here, we describe the capabilities of a newly configured combination of high-efficiency liquid chromatography and high-resolution/accuracy Fourier-transform mass spectrometry with electrospray ionization (LC-ESI-FTMS), designed for lipidomics applications that require the identification of PC in their lyso forms. The devised strategy, involving a separation by hydrophilic interaction liquid chromatography (HILIC) on spherical, fused-core ultrapure silica particles (2.7 μm) of a narrow-bore column (2.1 mm i.d.), enabled the identification of as many as 71 LPC species in the lipid extracts of gilthead sea bream (Sparus aurata) fillets. In this way, LPC as proton (43) and sodium (28) adducts, i.e., [M + H](+) and [M + Na](+) ions (with M representing the zwitterionic form), were identified. In several cases, the extremely high (sub-ppm) mass accuracy and the high chromatographic efficiency available with the adopted instrumentation enabled the distinction of isobaric and closely eluting LPC species. Informative tandem mass spectra, based on high-energy collision induced dissociation (HCD), were also obtained, thus distinguishing regioisomeric LPC species (i.e., LPC differing only for the location of the residual side chain on the glycerol backbone) and between proton and sodium adducts. Graphical Abstract Extracted Ion Current chromatogram (XIC) obtained for the m/z value 568.339, showing the presence of two regioisomeric Lysophosphatidylcholines. The corresponding high collisional energy tandem MS spectra, obtained using a HCD cell, are shown as insets.

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Validation of an LC–MS/MS method for the quantification of choline-related compounds and phospholipids in foods and tissues

Cited by 80 publications

References 28 publications

Metabolism and cytotoxic effects of phosphatidylcholine hydroperoxide in human hepatoma HepG2 cells

Metabolism and cytotoxic effects of phosphatidylcholine hydroperoxide in human hepatoma HepG2 cells

The PNPLA-family phospholipases involved in glycerophospholipid homeostasis of HeLa cells

Identification of isobaric lyso-phosphatidylcholines in lipid extracts of gilthead sea bream (Sparus aurata) fillets by hydrophilic interaction liquid chromatography coupled to high-resolution Fourier-transform mass spectrometry

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