Document Detail

High-performance liquid chromatographic assay with fluorescence detection for the evaluation of inhibitors against fatty acid amide hydrolase.
MedLine Citation:
PMID:  19484225     Owner:  NLM     Status:  MEDLINE    
A fluorescent assay for the evaluation of inhibitors of fatty acid amide hydrolase (FAAH) is described. Microsomes from rat brain served as enzyme source. N-(2-Hydroxyethyl)-4-pyren-1-ylbutanamide was designed and synthesized as novel fluorogenic substrate. For substrate solubilization, Triton X-100 was employed. The FAAH activity was determined directly without further sample clean-up by measuring the amount of 4-pyren-1-ylbutanoic acid released by the enzyme with reversed-phase HPLC and fluorescence detection. The known FAAH inhibitors URB597, phenyl hexanoyl oxazolopyridine (PHOP) and [6-(2-methyl-4,5-diphenyl-1H-imidazol-1-yl)hexyl]carbamic acid phenyl ester were used to validate the test assay.
Laura Forster; Alwine Schulze Elfringhoff; Matthias Lehr
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Publication Detail:
Type:  Journal Article; Validation Studies     Date:  2009-05-30
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  394     ISSN:  1618-2650     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2009 Jul 
Date Detail:
Created Date:  2009-06-24     Completed Date:  2009-08-27     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1679-85     Citation Subset:  IM    
Institute of Pharmaceutical and Medicinal Chemistry, University of Münster, Hittorfstrasse 58-62, 48149 Münster, Germany.
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MeSH Terms
Amidohydrolases / antagonists & inhibitors*,  metabolism
Brain / drug effects,  enzymology
Chromatography, High Pressure Liquid / methods*
Enzyme Inhibitors / chemistry,  pharmacology*
Fluorescent Dyes / analysis*,  chemical synthesis,  chemistry,  metabolism
Molecular Structure
Rats, Sprague-Dawley
Substrate Specificity
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Fluorescent Dyes; EC 3.5.-/Amidohydrolases; EC 3.5.1.-/fatty-acid amide hydrolase

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