HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sandhoff, R. Articles by Wieland, F. T. Search for Related Content PubMed PubMed Citation Articles by Sandhoff, R. Articles by Wieland, F. T. Social Bookmarking                      What's this?

The Journal of Lipid Research, Vol. 40, 126-132, January 1999
Copyright © 1999 by Lipid Research, Inc.

Original Article

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

Correspondence to: Roger Sandhoff

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.

Supplementary key words: membrane, lipid composition, ergosterol, stigmasterol, sterol, cholesterol-3-sulfate, internal standard, CHO cells, Golgi

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. Honda, K. Yamashita, H. Miyazaki, M. Shirai, T. Ikegami, G. Xu, M. Numazawa, T. Hara, and Y. Matsuzaki Highly sensitive analysis of sterol profiles in human serum by LC-ESI-MS/MS J. Lipid Res., September 1, 2008; 49(9): 2063 - 2073. [Abstract] [Full Text] [PDF]

				M.-S. Kuo, J. M. Kalbfleisch, P. Rutherford, D. Gifford-Moore, X.-d. Huang, R. Christie, K. Hui, K. Gould, and M. Rekhter Chemical analysis of atherosclerotic plaque cholesterol combined with histology of the same tissue J. Lipid Res., June 1, 2008; 49(6): 1353 - 1363. [Abstract] [Full Text] [PDF]

				X. Han, K. Yang, H. Cheng, K. N. Fikes, and R. W. Gross Shotgun lipidomics of phosphoethanolamine-containing lipids in biological samples after one-step in situ derivatization J. Lipid Res., July 1, 2005; 46(7): 1548 - 1560. [Abstract] [Full Text] [PDF]

				B. Brugger, C. Graham, I. Leibrecht, E. Mombelli, A. Jen, F. Wieland, and R. Morris The Membrane Domains Occupied by Glycosylphosphatidylinositol-anchored Prion Protein and Thy-1 Differ in Lipid Composition J. Biol. Chem., February 27, 2004; 279(9): 7530 - 7536. [Abstract] [Full Text] [PDF]

				E. Stuven, A. Porat, F. Shimron, E. Fass, D. Kaloyanova, B. Brugger, F. T. Wieland, Z. Elazar, and J. B. Helms Intra-Golgi Protein Transport Depends on a Cholesterol Balance in the Lipid Membrane J. Biol. Chem., December 26, 2003; 278(52): 53112 - 53122. [Abstract] [Full Text] [PDF]

				C. Eroglu, B. Brugger, F. Wieland, and I. Sinning Glutamate-binding affinity of Drosophila metabotropic glutamate receptor is modulated by association with lipid rafts PNAS, September 2, 2003; 100(18): 10219 - 10224. [Abstract] [Full Text] [PDF]

				X. Han and R. W. Gross Global analyses of cellular lipidomes directly from crude extracts of biological samples by ESI mass spectrometry: a bridge to lipidomics J. Lipid Res., June 1, 2003; 44(6): 1071 - 1079. [Abstract] [Full Text] [PDF]

				R. Sandhoff, S. T. Hepbildikler, R. Jennemann, R. Geyer, V. Gieselmann, R. L. Proia, H. Wiegandt, and H.-J. Grone Kidney Sulfatides in Mouse Models of Inherited Glycosphingolipid Disorders. DETERMINATION BY NANO-ELECTROSPRAY IONIZATION TANDEM MASS SPECTROMETRY J. Biol. Chem., May 31, 2002; 277(23): 20386 - 20398. [Abstract] [Full Text] [PDF]

				D. W. Johnson, H. J. ten Brink, and C. Jakobs A rapid screening procedure for cholesterol and dehydrocholesterol by electrospray ionization tandem mass spectrometry J. Lipid Res., October 1, 2001; 42(10): 1699 - 1706. [Abstract] [Full Text] [PDF]

				I. Gkantiragas, B. Brugger, E. Stuven, D. Kaloyanova, X.-Y. Li, K. Lohr, F. Lottspeich, F. T. Wieland, and J. B. Helms Sphingomyelin-enriched Microdomains at the Golgi Complex Mol. Biol. Cell, June 1, 2001; 12(6): 1819 - 1833. [Abstract] [Full Text] [PDF]

				B. Brugger, R. Sandhoff, S. Wegehingel, K. Gorgas, J. Malsam, J. B. Helms, W.-D. Lehmann, W. Nickel, and F. T. Wieland Evidence for Segregation of Sphingomyelin and Cholesterol during Formation of COPI-coated Vesicles J. Cell Biol., October 23, 2000; 151(3): 507 - 518. [Abstract] [Full Text] [PDF]

				R. Schneiter, B. Brugger, R. Sandhoff, G. Zellnig, A. Leber, M. Lampl, K. Athenstaedt, C. Hrastnik, S. Eder, G. Daum, et al. Electrospray Ionization Tandem Mass Spectrometry (ESI-MS/MS) Analysis of the Lipid Molecular Species Composition of Yeast Subcellular Membranes Reveals Acyl Chain-based Sorting/Remodeling of Distinct Molecular Species En Route to the Plasma Membrane J. Cell Biol., August 23, 1999; 146(4): 741 - 754. [Abstract] [Full Text] [PDF]

				K. A. Fagan, K. E. Smith, and D. M. F. Cooper Regulation of the Ca2+-inhibitable Adenylyl Cyclase Type VI by Capacitative Ca2+ Entry Requires Localization in Cholesterol-rich Domains J. Biol. Chem., August 18, 2000; 275(34): 26530 - 26537. [Abstract] [Full Text] [PDF]