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A highly sensitive RP-HPLC-fluorescence method to study aldehyde oxidase activity.
- Source :
-
Journal of AOAC International [J AOAC Int] 2011 Mar-Apr; Vol. 94 (2), pp. 550-4. - Publication Year :
- 2011
-
Abstract
- Aldehyde oxidase is a widely distributed enzyme that is involved in the metabolism of an extensive range of aldehydes and N-heterocyclic compounds with physiological, pharmacological, and toxicological relevance. In the present study, a highly sensitive RP-HPLC-fluorescence method based on the oxidation of phenanthridine to phenanthridinone has been developed and validated to assay aldehyde oxidase activity in biological samples. Determination of phenanthridinone was achieved on a C18 column using 10 mmol/L phosphate buffer (pH 5.0) containing 0.1 mmol/L EDTA-acetonitrile (40 + 60, v/v) as the mobile phase. The fluorescence intensity of phenanthridinone was measured at 364 nm with excitation at 236 nm. The proposed method was precise, accurate, specific and rapid (analysis time, approximately 8 min) with a mean RSD of 2.54%. Peak responses were linear from 0.5 to 100 nmol/L, with an LOD of 0.125 nmol/L. The applicability of the method was demonstrated by measurement of aldehyde oxidase activity in rat liver, kidney, ovary, and heart fractions.
- Subjects :
- Aldehyde Oxidase chemistry
Animals
Female
Kidney enzymology
Kidney metabolism
Liver enzymology
Liver metabolism
Male
Myocardium enzymology
Myocardium metabolism
Ovary enzymology
Ovary metabolism
Phenanthrenes chemistry
Phenanthrenes metabolism
Phenanthridines chemistry
Phenanthridines metabolism
Rats
Rats, Sprague-Dawley
Aldehyde Oxidase metabolism
Chromatography, High Pressure Liquid methods
Subjects
Details
- Language :
- English
- ISSN :
- 1060-3271
- Volume :
- 94
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of AOAC International
- Publication Type :
- Academic Journal
- Accession number :
- 21563690