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• W2046283668 abstract "We developed an improved sample extraction procedure that best prevents the acid-catalyzed breakdown of enol ether containing phospholipids and lysophospholipids compared with previously reported methods. The new method also results in high recovery of the major types of lysophospholipid species including LPA and LPI, which are generally more difficult to isolate than the more hydrophobic species. Breakdown of even 5–10% of the plasmenyl phospholipids is a significant problem if one is trying to assess the presence of lysophospholipids in a sample, as lysophospholipids are in relatively low abundance compared with plasmenyl phospholipids. It should also be pointed out that the new extraction method does result in a low level of plasmenyl phospholipid breakdown, thus, lysophospholipid analysis is best carried out by looking at a relative change in molecular species level as a result of a biological stimulus rather than looking at absolute levels in a single sample. Still, relative changes in lysophospholipids will be hard to spot if there is significant breakdown of plasmenyl phospholipid species; thus, the new extraction procedure is recommended in all studies of lysophospholipid abundance when using biological membranes that contain plasmenyl phospholipids. Analysis of lysophospholipids by the LC/ESI-MS/MS method described here allows for the detection of 20–1000 fmoles of molecular species using an instrument manufactured about a decade ago. With use of more updated instruments, it should be possible to extend the limit of quantification by a factor of ∼5- to 10-fold. Finally, we analyzed isobaric species of sn-1 ester and ether lysophospholipids and showed that the use of internal standards with fatty chains with odd carbon number, i.e., 17:0 or 17:1 chains is problematic and the use of d31-16:0 lysophospholipid internal standards is preferred. Also, LC is needed along with ESI-MS/MS to detect all the sn-1 ester and ether lysophospholipid with a high degree of confidence of species identification. The new method was used to quantify lysophospholipids present in mouse serum with acceptable inter-sample reproducibility. Finally, the method was used to illustrate that human X secreted phospholipase A2 added exogenously to HEK293 cells acts on a different membrane site than cells expressing endogenous phospholipase A2 within the cell." @default.
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• W2046283668 date "2010-02-01" @default.
• W2046283668 modified "2023-10-11" @default.
• W2046283668 title "Improved method for the quantification of lysophospholipids including enol ether species by liquid chromatography-tandem mass spectrometry" @default.
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• W2046283668 doi "https://doi.org/10.1194/jlr.d000885" @default.
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