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1.
Anal Chem ; 88(14): 6996-7004, 2016 07 19.
Article in English | MEDLINE | ID: mdl-27275841

ABSTRACT

Significant sensitivity enhancements in the tandem mass spectrometry-based analysis of complex mixtures of several phospholipid classes has been achieved via (13)C-TrEnDi. (13)C-TrEnDi-modified phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylcholine (PC) lipids extracted from HeLa cells demonstrated greater sensitivity via precursor ion scans (PISs) than their unmodified counterparts. Sphingomyelin (SM) species exhibited neither an increased nor decreased sensitivity following modification. The use of isotopically labeled diazomethane enabled the distinction of modified PE and modified PC species that would yield isobaric species with unlabeled diazomethane. (13)C-TrEnDi created a PE-exclusive PIS of m/z 202.1, two PS-exclusive PISs of m/z 148.1 and m/z 261.1, and a PIS of m/z 199.1 for PC species (observed at odd m/z values) and SM species (observed at even m/z values). The standardized average area increase after TrEnDi modification was 10.72-fold for PE species, 2.36-fold for PC, and 1.05-fold for SM species. The sensitivity increase of PS species was not quantifiable, as there were no unmodified PS species identified prior to derivatization. (13)C-TrEnDi allowed for the identification of 4 PE and 7 PS species as well as the identification and quantitation of an additional 4 PE and 4 PS species that were below the limit of detection (LoD) prior to modification. (13)C-TrEnDi also pushed 24 PE and 6 PC lipids over the limit of quantitation (LoQ) that prior to modification were above the LoD only.


Subject(s)
Diazomethane/chemistry , Phosphatidylcholines/analysis , Phosphatidylethanolamines/analysis , Phosphatidylserines/analysis , Carbon Isotopes , HeLa Cells , Humans , Limit of Detection , Methylation , Phosphatidylcholines/chemistry , Phosphatidylcholines/classification , Phosphatidylethanolamines/chemistry , Phosphatidylethanolamines/classification , Phosphatidylserines/chemistry , Phosphatidylserines/classification , Sensitivity and Specificity , Spectrometry, Mass, Electrospray Ionization/methods
2.
Lipids ; 39(1): 59-66, 2004 Jan.
Article in English | MEDLINE | ID: mdl-15055236

ABSTRACT

The FA composition of glycerophospholipid (GPL) classes and subclasses was investigated in whole animals of three marine bivalve mollusks: the Japanese oyster Crassostrea gigas, the blue mussel Mytilus edulis, and the Manila clam Ruditapes philippinarum. Individual organs (gills, mantle, foot, siphon, and muscle) of the Manila clam also were examined. The PS plasmalogen (PSplsm), PE plasmalogen (PEplsm), and PC plasmalogen (PCplsm) subclasses were isolated by HPLC, and their individual FA compositions were examined using GC. Plasmalogen forms of PS and PE, when compared to their respective diacyl forms, were found to be specifically enriched with non-methylene-interrupted (NMI) FA (7,15-22:2, 7,13-22:2, and their precursors) and 20:1 n-11 FA. Such a clear specific association was not found for PCplsm. Interestingly, this trend was most apparent in PSplsm, and the above FA were found to be, respectively, the predominant PUFA and monounsaturated FA in the PSplsm isolated from the three species. This specificity was maintained in all the analyzed organs of the Manila clam but varied in proportions: The highest level of plasmalogens, NMI FA, and 20:1 n-11 was measured in gills and the lowest was in muscle. These results represent the first comprehensive report on a FA composition of the PSplsm subclass isolated from mollusks. The fact that NMI FA and 20:1 n-11, which are thought to be biosynthesized FA, were mainly associated with aminophospholipid plasmalogens (PE and PS) is likely to have a functional significance in bivalve membranes.


Subject(s)
Bivalvia/chemistry , Fatty Acids/analysis , Ostreidae/chemistry , Plasmalogens/chemistry , Shellfish/analysis , Animals , Phosphatidylcholines/chemistry , Phosphatidylcholines/classification , Phosphatidylethanolamines/chemistry , Phosphatidylethanolamines/classification , Phosphatidylserines/chemistry , Phosphatidylserines/classification , Plasmalogens/classification
3.
Lipids ; 28(10): 945-7, 1993 Oct.
Article in English | MEDLINE | ID: mdl-27483556

ABSTRACT

A rapid method for the separation of the individual phospholipid classes phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI) by a single solid-phase extraction was developed. PC, PE, PS and PI were sequentially eluted from aminopropyl bonded silica with acetonitrile/n-propanol (2∶1, vol/vol), methanol, isopropanol/methanolic HCl (4∶1, vol/vol) and methanol/methanolic HCl (9∶1, vol/vol). Standard recoveries were over 95% for PC and PE and over 85% for PS and PI with undistorted fatty acid composition. The separation of complex lipid mixtures on aminopropyl minicolumns can be refined to the level of individual phospholipid classes.


Subject(s)
Phospholipids/analysis , Solid Phase Extraction/methods , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Phosphatidylcholines/analysis , Phosphatidylcholines/classification , Phosphatidylethanolamines/analysis , Phosphatidylethanolamines/classification , Phosphatidylinositols/analysis , Phosphatidylinositols/classification , Phosphatidylserines/analysis , Phosphatidylserines/classification , Phospholipids/classification
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