Page 101 - Pharma News Letter - Vol2 Issue 1

Research Publications

Investigating Fluorogenic Labelling Of Amine Coupled With
First-Order Derivative Spectrofluorimetry As A Versatile
Analytical Methodology: Application To Estimation Of
Benoxinate In Its Eye Drops

Spectrochimica Acta Part A: Molecular And Biomolecular Spectroscopy. 2024
Jan 15:305:123410.

------------------------------------------------------------------------------------
Rasha Aboshabana , Heba Samir Elama , Heba Elmansi , Samah Abo El Abass

Abstract

In this research, fluorogenic labelling followed by applying first-order derivative spectrofluorimetry
for the developed fluorophore is discussed as an alternative, sensitive and selective analytical
approach. Benoxinate, containing a primary amine and fluorescamine reagent were selected for
the study. Then the proposed methodology relies on the reaction between the primary amine in
benoxinate with fluorescamine that selectively reacts with primary amines to develop highly
fluorescent products. The fluorescamine-benoxinate developed fluorophore is identified by its
sharp first-order derivative peak at 465 nm following excitation at 386 nm in borate buffer,
pH 8. The optimum reaction conditions were ascertained. Following ICH validation guidelines,
the first order derivative of the relative fluorescence intensity for the developed fluorophore
was linearly related to benoxinate concentration and ranged from 20.0 to 200.0 ng/mL with
a detection limit of 3.36 ng/mL and a quantitation limit of 10.19 ng/mL, moreover, satisfying
accuracy and precision values were obtained upon statistical analysis of results. The offered
analytical method was successfully applied to quantify benoxinate in raw material and Benox®
eye drops as a direct application to a commercial formulation.

Reference:

https://doi.org/10.1016/j.saa.2023.123410

Vol. 2 - Issue 1 101