Rashidi Mohammad-Reza, Amini Kaveh, Khani Mohammad-Yaser, Faridi Akram, Hanaee Jalal, Sorouraddin Mohammad-Hossein
Tabriz University of Medical Sciences, School of Pharmacy, Department of Medicinal Chemistry, 51664-14766 Tabriz, Iran.
J AOAC Int. 2011 Mar-Apr;94(2):550-4.
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.
醛氧化酶是一种广泛分布的酶，参与多种具有生理、药理和毒理学意义的醛类和N-杂环化合物的代谢。在本研究中，已开发并验证了一种基于将菲啶氧化为菲啶酮的高灵敏度反相高效液相色谱-荧光法，用于测定生物样品中的醛氧化酶活性。使用含有0.1 mmol/L乙二胺四乙酸的10 mmol/L磷酸盐缓冲液（pH 5.0）-乙腈（40 + 60，v/v）作为流动相，在C18柱上测定菲啶酮。菲啶酮的荧光强度在激发波长为236 nm时于364 nm处测量。所提出的方法精确、准确、特异且快速（分析时间约8分钟），平均相对标准偏差为2.54%。峰响应在0.5至100 nmol/L范围内呈线性，检测限为0.125 nmol/L。通过测量大鼠肝脏、肾脏、卵巢和心脏组分中的醛氧化酶活性，证明了该方法的适用性。
