High-performance liquid chromatographic assay with fluorescence detection for the evaluation of inhibitors against fatty acid amide hydrolase
| Autor: | Laura Forster, Matthias Lehr, Alwine Schulze Elfringhoff |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Biochemistry
High-performance liquid chromatography Fluorescence spectroscopy Amidohydrolases Substrate Specificity Analytical Chemistry Rats Sprague-Dawley chemistry.chemical_compound Carbamic acid Fatty acid amide hydrolase Animals Enzyme Inhibitors Chromatography High Pressure Liquid Fluorescent Dyes chemistry.chemical_classification Chromatography Molecular Structure Brain Substrate (chemistry) URB597 Rats Enzyme chemistry Microsome Female |
| Zdroj: | Analytical and Bioanalytical Chemistry. 394:1679-1685 |
| ISSN: | 1618-2650 1618-2642 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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