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Analysis and Experimental Inhibition of Distal Cholesterol Biosynthesis

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Abstract

During the last decade, our understanding about the function and biological activity of metabolites has drastically changed. Metabolites previously seen as sole bystanders of biochemical reactions without unique activity are becoming more and more recognized for their biological functions. One class of metabolites for which such a paradigm shift has become evident is cholesterol and its precursors. Here, we will give a brief overview about the post-squalene pathway of cholesterol biosynthesis, its inhibition, the biological functions of its intermediates and put special emphasis on the analysis of the occurring metabolites. We will compare liquid chromatography and gas chromatography-based analysis platforms, describe mass spectrometric fragmentations and explain different derivatization strategies. In addition, we will present a full spectral data set of 27 analytes of which 24 are sterols, synthesized, isolated and characterized in several studies in our laboratories. Finally, with our overview, we hope to assist researchers in the field of cholesterol biosynthesis with the chemical analysis of cholesterol and its physiological as well as non-physiological precursors.

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Abbreviations

7-DHC:

7-Dehydrocholesterol

ABS:

Antley–Bixler Syndrome

APCI:

Atmospheric pressure chemical ionization

BSTFA:

N,O-Bis(trimethylsilyl)trifluoroacetamide

DMES:

Dimethylethylsilyl

DTE:

1,4-Dithioerythritol

ECNI:

Electron capture negative ionization

EI:

Electron ionization

ER:

Estrogen receptor

ESI:

Electrospray ionization

FAME:

Fatty acid methyl ester

FF-MAS:

Follicular fluid meiosis-activating sterol

FID:

Flame ionization detection

GC:

Gas chromatography

GC–MS(MS):

Gas chromatography (tandem) mass spectrometry

IT:

Ion trap

LC:

Liquid chromatography

LC–MS(MS):

Liquid chromatography (tandem) mass spectrometry

LXR:

Liver X receptor

MSTFA:

N-Methyl-N-trimethylsilyltrifluoroacetamide

MtBSTFA:

N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide

ODS:

Octadecylsilyl

PCI:

Positive chemical ionization

RRT:

Relative retention time

SC:

Side chain

SLOS:

Smith–Lemli–Opitz syndrome

SPE:

Solid-phase extraction

SIM:

Selected ion monitoring

SIS:

Single ion storage

SREBP:

Sterol response element-binding protein

SRM:

Selected reaction monitoring

tBDMS:

tert-Butyl dimethylsilyl

tBME:

tert-Butyl methyl ether

T-MAS:

Testis meiosis-activating sterol

TMCS:

Trimethylchlorosilane

TMIS:

Trimethyliodosilane

TMS:

Trimethylsilyl

TMSiOH:

Trimethylsilanol

TSIM:

N-(Trimethylsilyl)imidazole

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Authors and Affiliations

  1. Leiden University Medical Center, Center for Proteomics and Metabolomics, Albinusdreef 2, 2300 RC, Leiden, The Netherlands

    Martin Giera

  2. Department für Pharmazie-Zentrum für Pharmaforschung, Ludwig-Maximilians-Universität, Butenandtstrasse 5-13, 81377, Munich, Germany

    Christoph Müller & Franz Bracher

Authors
  1. Martin Giera
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  2. Christoph Müller
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  3. Franz Bracher
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Correspondence to Martin Giera.

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Published in the topical collection Recent Developments in Clinical Omics with guest editors Martin Giera and Manfred Wuhrer.

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Giera, M., Müller, C. & Bracher, F. Analysis and Experimental Inhibition of Distal Cholesterol Biosynthesis. Chromatographia 78, 343–358 (2015). https://doi.org/10.1007/s10337-014-2796-4

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