Application of different spectrofluorimetric methods for determination of lesinurad and allopurinol in pharmaceutical preparation and human plasma.

Lesinurad and allopurinol combination is newly FDA approved for treatment of patients suffering from hyperuricemia associated with uncontrolled gout. In the present work, two different highly sensitive, selective and accurate fluorescence spectroscopic methods were developed for quantitative analysis of lesinurad and allopurinol in their pharmaceutical dosage form without any tedious operation procedure. Lesinurad was quantitatively analyzed based on its unique native fluorescence nature. Lesinurad fluorescence emission was quantitatively determined at 343 nm after excitation at 288 nm without any interference from allopurinol. Allopurinol, has free terminal secondary amino group, reacted with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBDCl) through nucleophilic substitution mechanism forming highly fluorescent dark yellow fluorophore. Allopurinol was quantitavely analyzed based on measurement the emission fluorescence intensity of the fluorescent dark yellow fluorophore at 535 nm after excitation at 465 nm. Different parameters which affect the described methods of the studied drugs were carefully checked and optimized. Calibration graphs were found to be linear over the concentration range of 0.25-4.0 μg/mL for lesinurad and 0.2-20 μg/mL for allopurinol. The proposed methods were successfully applied for the quantitative analysis of the two drugs in Duzallo® pharmaceutical dosage form and spiked human plasma.