Development of a Sensitive Spectrofluorometric-Multivariate Calibration Method for Enzyme Kinetic of Aldehyde Oxidase

Mohammad Hossein Sorouraddin; Ebrahim Fooladi; Abdolhossein Naseri; Mohammad-Reza Rashidi

doi:10.22037/ijpr.2010.807

IJ Pharmaceutical Research

Current Issue All Issues In Press Accepted Manuscripts Search

Journal Information Editors & Boards Indexing and Listing Sources Journal Metrics Open Peer Review (OPR)Publication Ethics and Malpractice Statement Reviewer and AE Registration Form Contact Us Support

APC

Authors Guide Submit Manuscript

https://doi.org/10.22037/ijpr.2010.807

Development of a Sensitive Spectrofluorometric-Multivariate Calibration Method for Enzyme Kinetic of Aldehyde Oxidase

Author(s):

Mohammad Hossein Sorouraddin¹,

Ebrahim Fooladi¹,

Abdolhossein Naseri²,

Mohammad-Reza Rashidi^3,*

1Department of Analytical Chemistry, Faculty of Chemistry, Tabriz University, Tabriz, Iran

2Applied Chemistry Department, Faculty of Science, Islamic Azad University, Tabriz branch, Tabriz, Iran

3Department of Medicinal Chemistry, School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

IJ Pharmaceutical Research:Vol. 8, issue 3; 169-177

Published online:Nov 20, 2010

Article type:Research Article

Received:Jul 01, 2008

Accepted:Nov 01, 2008

How to Cite:Mohammad Hossein SorouraddinEbrahim FooladiAbdolhossein NaseriMohammad-Reza RashidiDevelopment of a Sensitive Spectrofluorometric-Multivariate Calibration Method for Enzyme Kinetic of Aldehyde Oxidase.Iran J Pharm Res.8(3):e128634.https://doi.org/10.22037/ijpr.2010.807.

Abstract

Attempts to obtain experimental values for the kinetic parameters of phenanthridine oxidation by guinea pig or rabbit liver aldehyde oxidase using common spectrophotometric methods have not been successful due to a lower limit of detection. In the present study, a new spectrofluorimetric assay in combination with a multivariate calibration method for enzymatic kinetic study of aldehyde oxidase activity, using phenanthridine as the substrate, has been developed and validated.

Phenanthridine and phenanthridinone binary mixtures were prepared in a dynamic linear range of 0.025-1 µM and the emission flourimetric spectra of the solutions recorded at the excitation and emission wavelengths of 236 and 320-450 nm, respectively. The optimized calibration model of partial least squares (PLS) method was applied for the simultaneous determination of the concentration of each chemical in the prediction set. The limits of detection for phenanthridine and phenanthridinone were found to be 2.13 ± 0.33 and 3.41 ± 0.34 nM (mean ± SD, n = 5), respectively. This method was then used for kinetic study of phenanthridine oxidation using guinea pig and rat hepatic aldehyde oxidase. The results were compared with those obtained from a univariate spectroscopic method.

Using this new spectrofluorimetric-multivariate calibration method, the K_m value for the oxidation of phenanthridine with guinea pig and rat liver aldehyde oxidase were obtained as 0.83 ± 0.08 and 2.20 ± 0.40, µM (mean ± SD, n = 3), respectively.

Keywords

comments

Crossmark

Checking

Share on

Comments

Number of Comments:0

Cited by

CrossRef 0

Metrics

Get Permission (article level)

Purchasing Reprints

Copyright Clearance Center (CCC) handles bulk orders for article reprints for Brieflands. To place an order for reprints, please click here ( https://www.copyright.com/landing/reprintsinquiryform/ ). Clicking this link will bring you to a CCC request form where you can provide the details of your order. Once complete, please click the ‘Submit Request’ button and CCC’s Reprints Services team will generate a quote for your review.

Search Relations

Author(s):

Mohammad Hossein Sorouraddin:[PubMed][Scholar]
Ebrahim Fooladi:[PubMed][Scholar]
Abdolhossein Naseri:[PubMed][Scholar]
Mohammad-Reza Rashidi:[PubMed][Scholar]