Combined gas chromatographic/mass spectrometric analysis of cholesterol precursors and plant sterols in cultured cells

Jure Acimovic, Anita Lövgren-Sandblom, K. Monostory, Damjana Rozman, Marko Golicnik, Dieter Lutjohann, Ingemar Björkhem

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

We developed a powerful gas chromatographic/mass spectrometric method allowing quantitative analysis of 11 structurally similar cholesterol precursors and plant sterols (squalene, desmosterol, 7-dehydrocholesterol, lathosterol, zymosterol, dihydro-lanosterol, lanosterol, FF-MAS, T-MAS, campesterol, sitosterol) from cultured human hepatocytes in a single chromatographic run. Deuterium labelled cholesterol, sitosterol and lathosterol were used as internal standards. Care was taken to select ions for the detection that gave the most appropriate discrimination in the assay. Replicate analyses gave a coefficient of variation less than 6%. Recovery experiments were satisfactory for 7-dehydrocholesterol, campesterol, desmosterol, lathosterol, zymosterol and cholesterol with less than 7% difference between expected and found levels. For other sterols, the difference between expected and found levels varied between 10 and 16%. It is concluded that this method is suitable for studies on the effect of different inhibitors and stimulators of cholesterol synthesis in cultured cells. Additionally, the method is relevant also for clinical applications since abnormally increased late cholesterol intermediates in patients are representations of the inherited disorders linked to different enzyme defects in the post-squalene cholesterol biosynthesis.

Original languageEnglish
Pages (from-to)2081-2086
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume877
Issue number22
DOIs
Publication statusPublished - Jul 15 2009

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Phytosterols
Cultured Cells
Gases
Cholesterol
Cells
Desmosterol
Lanosterol
Squalene
Anticholesteremic Agents
Deuterium
Sterols
Biosynthesis
Hepatocytes
Assays
Ions
Recovery
Enzymes
Defects
lathosterol
Chemical analysis

Keywords

  • Cholesterol biosynthesis intermediates
  • Gas chromatography/mass spectrometry
  • Human hepatocytes

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Cell Biology
  • Clinical Biochemistry

Cite this

Combined gas chromatographic/mass spectrometric analysis of cholesterol precursors and plant sterols in cultured cells. / Acimovic, Jure; Lövgren-Sandblom, Anita; Monostory, K.; Rozman, Damjana; Golicnik, Marko; Lutjohann, Dieter; Björkhem, Ingemar.

In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 877, No. 22, 15.07.2009, p. 2081-2086.

Research output: Contribution to journalArticle

Acimovic, Jure ; Lövgren-Sandblom, Anita ; Monostory, K. ; Rozman, Damjana ; Golicnik, Marko ; Lutjohann, Dieter ; Björkhem, Ingemar. / Combined gas chromatographic/mass spectrometric analysis of cholesterol precursors and plant sterols in cultured cells. In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2009 ; Vol. 877, No. 22. pp. 2081-2086.
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