Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry
A mass spectrometric method for the quantification of free cholesterol in cells and subcellular membranes is presented. The method is based on a simple one-step chemical derivatization of cholesterol to cholesterol-3-sulfate by a sulfur trioxide-pyridine complex. Quantification is performed by nano-...
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1999-01-01
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doaj-914589034f8a4a3ea28cfbb1bdf4b05a2021-04-26T05:46:37ZengElsevierJournal of Lipid Research0022-22751999-01-01401126132Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometryRoger Sandhoff0Britta Brügger1Dieter Jeckel2Wolf D. Lehmann3Felix T. Wieland4To whom correspondence should be addressed; Biochemie-Zentrum Heidelberg (BZH), University of Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, GermanyBiochemie-Zentrum Heidelberg (BZH), University of Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, GermanyBiochemie-Zentrum Heidelberg (BZH), University of Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, GermanyCentral Spectroscopy Department, German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, GermanyBiochemie-Zentrum Heidelberg (BZH), University of Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, GermanyA mass spectrometric method for the quantification of free cholesterol in cells and subcellular membranes is presented. The method is based on a simple one-step chemical derivatization of cholesterol to cholesterol-3-sulfate by a sulfur trioxide-pyridine complex. Quantification is performed by nano-electrospray ionization tandem mass spectrometry (nanoESI-MS/MS) using a stable isotope labeled internal standard. The determination of free cholesterol is demonstrated in about 250 cells of a Chinese hamster ovary (CHO) cell line. With this method a molar ratio of free cholesterol to total phospholipids of 0.34 mol/mol in CHO cells was determined. In a subcellular membrane fraction enriched in Golgi membranes, a molar ratio of free cholesterol to total phospholipids of 0.57 mol/mol was determined. The method should be of value for quantification of other sterols as demonstrated for ergosterol and stigmasterol.—Sandhoff, R., B. Brügger, D. Jeckel, W. D. Lehmann, and F. T. Wieland. Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry. J. Lipid Res. 1999. 40: 126–132.http://www.sciencedirect.com/science/article/pii/S0022227520333472membranelipid compositionergosterolstigmasterolsterolcholesterol-3-sulfate |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Roger Sandhoff Britta Brügger Dieter Jeckel Wolf D. Lehmann Felix T. Wieland |
spellingShingle |
Roger Sandhoff Britta Brügger Dieter Jeckel Wolf D. Lehmann Felix T. Wieland Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry Journal of Lipid Research membrane lipid composition ergosterol stigmasterol sterol cholesterol-3-sulfate |
author_facet |
Roger Sandhoff Britta Brügger Dieter Jeckel Wolf D. Lehmann Felix T. Wieland |
author_sort |
Roger Sandhoff |
title |
Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
title_short |
Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
title_full |
Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
title_fullStr |
Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
title_full_unstemmed |
Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
title_sort |
determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
1999-01-01 |
description |
A mass spectrometric method for the quantification of free cholesterol in cells and subcellular membranes is presented. The method is based on a simple one-step chemical derivatization of cholesterol to cholesterol-3-sulfate by a sulfur trioxide-pyridine complex. Quantification is performed by nano-electrospray ionization tandem mass spectrometry (nanoESI-MS/MS) using a stable isotope labeled internal standard. The determination of free cholesterol is demonstrated in about 250 cells of a Chinese hamster ovary (CHO) cell line. With this method a molar ratio of free cholesterol to total phospholipids of 0.34 mol/mol in CHO cells was determined. In a subcellular membrane fraction enriched in Golgi membranes, a molar ratio of free cholesterol to total phospholipids of 0.57 mol/mol was determined. The method should be of value for quantification of other sterols as demonstrated for ergosterol and stigmasterol.—Sandhoff, R., B. Brügger, D. Jeckel, W. D. Lehmann, and F. T. Wieland. Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry. J. Lipid Res. 1999. 40: 126–132. |
topic |
membrane lipid composition ergosterol stigmasterol sterol cholesterol-3-sulfate |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520333472 |
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