title: A highly sensitive RP-HPLC-fluorescence method to study aldehyde oxidase activity.
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.
authors.affiliation: Tabriz University of Medical Sciences, School of Pharmacy, Department of Medicinal Chemistry, 51664-14766 Tabriz, Iran. rashidi@tbzmed.ac.ir
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Rashidi
authors.forename: Kaveh
authors.initials: K
authors.lastname: Amini
authors.forename: Mohammad-Yaser
authors.initials: MY
authors.lastname: Khani
authors.forename: Akram
authors.initials: A
authors.lastname: Faridi
authors.forename: Jalal
authors.initials: J
authors.lastname: Hanaee
authors.forename: Mohammad-Hossein
authors.initials: MH
authors.lastname: Sorouraddin
mesh_terms: D042931:Aldehyde Oxidase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D007668:Kidney / Q000201:enzymology / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008297:Male; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism; D010053:Ovary / Q000201:enzymology / Q000378:metabolism; D010616:Phenanthrenes / Q000737:chemistry / Q000378:metabolism; D010617:Phenanthridines / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D010616:Phenanthrenes; D010617:Phenanthridines; C040601:phenanthridone; D042931:Aldehyde Oxidase