Abstract
Objective: The aim of the method was to develop a novel, simple and rapid fluorometric method determination of Amiloride (AMD) in pure form and pharmaceutical drugs via fluorescence measurements.
 Methods: The method depends on fluorescence quenching of 2H-chromene azo dye (2-(4-nitrophenyl)-N-(4-(phenyldiazenyl)-2H-chromen-4-amine) upon adding Amiloride (AMD) using homemade ISNAG 2 X 90° multi solar cell via low-pressure mercury lamp at two significant wavelengths 184.9 and 253.7 nm combined with continuous flow injection analysis.
 Results: Under the optimized conditions, the fluorescence quenching linear working range and percentage linearity (r2%) was (0.03-8 mmol/l) and 98.78 %, respectively. The suggested method was effectively applied to the determination of AMD in two different pharmaceutical drugs and compared with the classical method (UV-vis spectrophotometry at λ=540 nm).
 Conclusion: The proposed and established method is simple, direct, and efficient. The statistical comparison results using a t-test at 95% confidence interval that was applied to compare the new and classical method showed there are no significant differences between the two methods.
Highlights
Amiloride (AMD) [chemically it is known as [3,5-diamino-N(diaminomethylene)-6-chloropyrazinecarboxamide] is a pyrazine carbonyl guanidine derivative
Several analytical techniques have been reported for Amiloride determination, including spectrophotometry [8,9,10], high-performance liquid chromatography (HPLC)[11, 12], Atomic Emission spectrometry [13] and polarography [14]
A set of 2H-chromene azo dye concentration ranging from 0.05-0.5 mmol/l were prepared as a carrier stream with a flow rate of 1.2 ml/min and 50 μl of 4 mmol/l of amiloride as an injected sample to study the effect of the amiloride solution on the quenching of continuous azo dye fluorescence
Summary
Amiloride (AMD) [chemically it is known as [3,5-diamino-N(diaminomethylene)-6-chloropyrazinecarboxamide] is a pyrazine carbonyl guanidine derivative (fig. 1). Several analytical techniques have been reported for Amiloride determination, including spectrophotometry [8,9,10], high-performance liquid chromatography (HPLC)[11, 12], Atomic Emission spectrometry [13] and polarography [14]
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