A highly sensitive method for analysis of 7-dehydrocholesterol for the study of smith-lemli-opitz syndrome

Wei Liu, Libin Xu, Connor Lamberson, Dorothea Haas, Zeljka Korade, Ned A. Porter

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Abstract We describe a highly sensitive method for the detection of 7-dehydrocholesterol (7-DHC), the biosynthetic precursor of cholesterol, based on its reactivity with 4-phenyl- 1,2,4-triazoline-3,5-dione (PTAD) in a Diels-Alder cycloaddition reaction. Samples of biological tissues and fl uids with added deuterium-labeled internal standards were derivatized with PTAD and analyzed by LC-MS. This protocol permits fast processing of samples, short chromatography times, and high sensitivity. We applied this method to the analysis of cells, blood, and tissues from several sources, including human plasma. Another innovative aspect of this study is that it provides a reliable and highly reproducible measurement of 7-DHC in 7-dehydrocholesterol reductase ( Dhcr7 )-HET mouse (a model for Smith-Lemli-Opitz syndrome) samples, showing regional differences in the brain tissue. We found that the levels of 7-DHC are consistently higher in Dhcr7 -HET mice than in controls, with the spinal cord and peripheral nerve showing the biggest differences. In addition to 7-DHC, sensitive analysis of desmosterol in tissues and blood was also accomplished with this PTAD method by assaying adducts formed from the PTAD ";ene" reaction. The method reported here may provide a highly sensitive and high throughput way to identify at-risk populations having errors in cholesterol biosynthesis. -Liu, W., L. Xu, C. Lamberson, D. Haas, Z. Korade, and N. A. Porter. A highly sensitive method for analysis of 7-dehydrocholesterol for the study of Smith- Lemli-Opitz syndrome.

Original languageEnglish (US)
Pages (from-to)329-337
Number of pages9
JournalJournal of Lipid Research
Volume55
Issue number2
DOIs
StatePublished - Feb 1 2014

Fingerprint

Smith-Lemli-Opitz Syndrome
Tissue
Cycloaddition Reaction
Blood
Desmosterol
Cholesterol
Plasma (human)
Spinal Nerves
Cycloaddition
Deuterium
Biosynthesis
Chromatography
Peripheral Nerves
Brain
Blood Cells
Spinal Cord
Throughput
7-dehydrocholesterol
Processing

Keywords

  • 7-dehydrocholesterol reductase
  • Desmosterol
  • Diels-Alder cycloaddition
  • Gas chromatography
  • Liquid chromatography-mass spectrometry
  • PTAD

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

A highly sensitive method for analysis of 7-dehydrocholesterol for the study of smith-lemli-opitz syndrome. / Liu, Wei; Xu, Libin; Lamberson, Connor; Haas, Dorothea; Korade, Zeljka; Porter, Ned A.

In: Journal of Lipid Research, Vol. 55, No. 2, 01.02.2014, p. 329-337.

Research output: Contribution to journalArticle

Liu, Wei ; Xu, Libin ; Lamberson, Connor ; Haas, Dorothea ; Korade, Zeljka ; Porter, Ned A. / A highly sensitive method for analysis of 7-dehydrocholesterol for the study of smith-lemli-opitz syndrome. In: Journal of Lipid Research. 2014 ; Vol. 55, No. 2. pp. 329-337.
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