Document Detail


In vivo D2O labeling to quantify static and dynamic changes in cholesterol and cholesterol esters by high resolution LC/MS.
MedLine Citation:
PMID:  20884843     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
High resolution LC/MS-MS and LC/APPI-MS methods have been established for the quantitation of flux in the turnover of cholesterol and cholesterol ester. Attention was directed toward quantifying the monoisotopic mass (M0) and that of the singly deuterated labeled (M+1) isotope. A good degree of isotopic dynamic range has been achieved by LC/MS-MS ranging from 3-4 orders of magnitude. Correlation between the linearity of GC/MS and LC atmospheric pressure photoionization (APPI)-MS are complimentary (r² = 0.9409). To prove the viability of this particular approach, male C57Bl/6 mice on either a high carbohydrate (HC) or a high fat (HF) diet were treated with ²H₂O for 96 h. Gene expression analysis showed an increase in the activity of stearoyl-CoA desaturase (Scd1) in the HC diet up to 69-fold (P < 0.0008) compared with the HF diet. This result was supported by the quantitative flux measurement of the isotopic incorporation of ²H into the respective cholesterol and cholesterol ester (CE) pools. We concluded that it is possible to readily obtain static and dynamic measurement of cholesterol and CEs in vivo by coupling novel LC/MS methods with stable isotope-based protocols.
Authors:
Jose Castro-Perez; Stephen F Previs; David G McLaren; Vinit Shah; Kithsiri Herath; Gowri Bhat; Douglas G Johns; Sheng-Ping Wang; Lyndon Mitnaul; Kristian Jensen; Robert Vreeken; Thomas Hankemeier; Thomas P Roddy; Brian K Hubbard
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Publication Detail:
Type:  Journal Article     Date:  2010-09-30
Journal Detail:
Title:  Journal of lipid research     Volume:  52     ISSN:  0022-2275     ISO Abbreviation:  J. Lipid Res.     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2010-12-14     Completed Date:  2011-06-21     Revised Date:  2012-01-02    
Medline Journal Info:
Nlm Unique ID:  0376606     Medline TA:  J Lipid Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  159-69     Citation Subset:  IM    
Affiliation:
Atherosclerosis Exploratory Biomarkers Group, Merck & Co., Inc., Rahway, NJ, USA. jose_castro-perez@merck.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Cholesterol / analysis*,  blood,  metabolism
Cholesterol Esters / analysis*,  metabolism
Chromatography, Liquid / methods*
Deuterium Oxide / chemistry*
Isotopes
Male
Mass Spectrometry / methods*
Mice
Mice, Inbred C57BL
Palmitates / blood
Stearoyl-CoA Desaturase / metabolism
Chemical
Reg. No./Substance:
0/Cholesterol Esters; 0/Isotopes; 0/Palmitates; 57-88-5/Cholesterol; 7789-20-0/Deuterium Oxide; EC 1.14.19.1/Stearoyl-CoA Desaturase

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