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Determination of cholesterol at the low picomole level by nano-electrospray ionization tandem mass spectrometry.

Authors
  • Sandhoff, R
  • Brügger, B
  • Jeckel, D
  • Lehmann, W D
  • Wieland, F T
Type
Published Article
Journal
Journal of lipid research
Publication Date
Jan 01, 1999
Volume
40
Issue
1
Pages
126–132
Identifiers
PMID: 9869658
Source
Medline
License
Unknown

Abstract

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.

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