| An improved method for quantification of cholesterol and cholesteryl esters in human monocyte-derived macrophages by high performance liquid chromatography with identification of unassigned cholesteryl ester species by means of secondary ion mass spectrometry. | |
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MedLine Citation:
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PMID: 9162758 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The measurement of cholesteryl esters in human monocyte-derived macrophages using previously described high performance liquid chromatography methods is hampered by the presence in these cells of large amounts of triglycerides. We present a simple reversed phase high performance liquid chromatography protocol for quantification of cholesterol and cholesteryl esters in human monocyte/macrophages or other triglyceride-rich cells. Our method requires only lipid extraction and hydrolysis of triglycerides using a solution of ethanolic potassium hydroxide and is of sufficient sensitivity to allow measurement in 10(5) cells. Use of this protocol led to the isolation of eight previously unassigned cholesteryl ester peaks comprising 16% of the total cholesteryl ester content of human monocyte-derived macrophages. Using time-of-light secondary ion mass spectrometry and synthesized authentic standards, seven of these peaks were found to comprise cholesterol esterified with polyunsaturated n-3 (omega 3) (cholesteryl eicosapentaenoate, docosatrienoate, docosapentaenoate, and docosahexaenoate) and n-6 (omega 6) (cholesteryl docosatetraenoate, eicosadienoate, and eicosatrienoate) fatty acids. The remaining peak was shown to be the cholesteryl ester of n-7 (omega 7) palmitoleic acid by comparison with a commercially available standard. The identification of all the cholesteryl esters in cholesterol-loaded human monocyte-derived macrophages will assist future studies of lipid metabolism in these cells. |
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Authors:
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P Cullen; M Fobker; K Tegelkamp; K Meyer; F Kannenberg; A Cignarella; A Benninghoven; G Assmann |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of lipid research Volume: 38 ISSN: 0022-2275 ISO Abbreviation: J. Lipid Res. Publication Date: 1997 Feb |
Date Detail:
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Created Date: 1997-05-27 Completed Date: 1997-05-27 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0376606 Medline TA: J Lipid Res Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 401-9 Citation Subset: IM |
Affiliation:
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Institut für Arterioskleroseforschung, Westfälische Wilhelms-Universität Münster, Germany. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Cells, Cultured Cholesterol / analysis*, chemistry Cholesterol Esters / analysis*, chemistry Chromatography, High Pressure Liquid Ethanol / chemistry Gas Chromatography-Mass Spectrometry Humans Hydroxides / chemistry Linear Models Macrophages / chemistry* Mice Monocytes / chemistry* Potassium Compounds / chemistry Reproducibility of Results Sensitivity and Specificity Spectrometry, Mass, Secondary Ion Spectrophotometry, Ultraviolet |
| Chemical | |
Reg. No./Substance:
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0/Cholesterol Esters; 0/Hydroxides; 0/Potassium Compounds; 1310-58-3/potassium hydroxide; 57-88-5/Cholesterol; 64-17-5/Ethanol |
| Comments/Corrections | |
Erratum In:
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J Lipid Res 1998 May;39(5):1132 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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