Profiling and imaging ion mobility-mass spectrometry analysis of cholesterol and 7-dehydrocholesterol in cells via sputtered silver MALDI

Libin Xu, Michal Kliman, Jay G. Forsythe, Zeljka Korade, Anthony B. Hmelo, Ned A. Porter, John A. McLean

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Profiling and imaging of cholesterol and its precursors by mass spectrometry (MS) are important in a number of cholesterol biosynthesis disorders, such as in Smith-Lemli-Opitz syndrome (SLOS), where 7-dehydrocholesterol (7-DHC) is accumulated in affected individuals. SLOS is caused by defects in the enzyme that reduces 7-DHC to cholesterol. However, analysis of sterols is challenging because these hydrophobic olefins are difficult to ionize for MS detection. We report here sputtered silver matrix-assisted laser desorption/ionization (MALDI)-ion mobility-MS (IM-MS) analysis of cholesterol and 7-DHC. In comparison with liquid-based AgNO3 and colloidal Ag nanoparticle (AgNP), sputtered silver NP (10-25 nm) provided the lowest limits-of-detection based on the silver coordinated [cholesterol∈+∈Ag]+ and [7-DHC∈+∈Ag]+ signals while minimizing dehydrogenation products ([M∈+∈Ag-2H]+). When analyzing human fibroblasts that were directly grown on poly-L-lysine-coated ITO glass plates with this technique, in situ, the 7-DHC/cholesterol ratios for both control and SLOS human fibroblasts are readily obtained. The m/z of 491 (specific for [7-DHC∈+∈107Ag]+) and 495 (specific for [cholesterol∈+∈109Ag]+) were subsequently imaged using MALDI-IM-MS. MS images were co-registered with optical images of the cells for metabolic ratio determination. From these comparisons, ratios of 7-DHC/cholesterol for SLOS human fibroblasts are distinctly higher than in control human fibroblasts. Thus, this strategy demonstrates the utility for diagnosing/assaying the severity of cholesterol biosynthesis disorders in vitro. Graphical Abstract [Figure not available: see fulltext.]

Original languageEnglish (US)
Pages (from-to)924-933
Number of pages10
JournalJournal of the American Society for Mass Spectrometry
Volume26
Issue number6
DOIs
StatePublished - Jun 1 2015
Externally publishedYes

Keywords

  • 7-Dehydrocholesterol
  • Cholesterol
  • Imaging
  • Ion mobility-mass spectrometry
  • Silver
  • Smith-Lemli-Opitz syndrome
  • Sputter coating

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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