November-December 2023 Indian Journal of Pharmaceutical Sciences 1660 Research Paper *Address for correspondence E-mail: prashant.hanamshetty@gmail.com Hanamshetty et al.: Spectrophotometric Determination of Amiloride The present study proposes two straightforward, responsive and inveterate spectrophotometric techniques for the detection of amiloride hydrochloride. The methodologies are mainly relay on the generation of charge transfer complex, with the reagents 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (Method A) and p-chloranilic acid (Method B), hence an enduring 1:1 stoichiometric complex were accomplished, which are yellow and pink colour correspondingly. The generated chromogens optical densities were measured at 476 and 508 nm respectively. Beer’s law been tested in the range of 5-30 µg.ml -1 and 1-6 µg.ml -1 . The other analytical parameters like, Limit of Detection (LOD), Limit of Quantification (LOQ), accuracy and precisions were examined. The mentioned approaches were successfully adopted for the determination of the amiloride hydrochloride in tablet formulation with high accuracy, better recoveries and acceptable relative standard deviation. Key words: Method validation, amiloride hydrochloride, 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, p-chloranilic acid, spectrophotometry, charge transfer complex Chemically Amiloride, 3,5-diamino-6-chloro- N-(diaminomethylene) pyrazine-2-carboxamide (fig. 1), which operates through blocking the epithelial sodium channel through decreasing sodium reabsorption in the distal convoluted tubules and collecting ducts in the kidneys [1] . This stimulates the elimination of salt and water from the body, but without depleting potassium. The medicine is typically used in combination with thiazide (eg. co-amilozide) or loop diuretics (eg. co-amilofruse) (e.g. co-amilofruse). Due to its potassium-sparing qualities, hyperkalemia (high blood potassium levels) are sometimes recorded in patients on Amiloride. It is used in the therapy of hypertension and congestive heart failure. It is beneficial for the prevention of hypokalemia generated by hydrochlorothiazide especially during extended therapy. Amiloride is available on the market as combination medicine with furosemide such as Amifru tab, Amimide, Exna-k tab. It is also available in conjunction with Atenelol and Hydrochlorothiazide namely Beta-Biduret cap, BP-Loride tab, Hipres-D cap. Spectrophotometric Assay Methods for Determination of Amiloride: A Charge-Transfer Reaction Approach with 2,3-Dichloro-5,6-Dicyano-1,4-Benzoquinone and p-Chloranilic Acid Reagents P. C. HANAMSHETTY*, K. SIDDAPPA 1 , P. CHAVAN 2 AND M. M. NAGABHUSHANA 3 Department of Chemistry, Guru Nanak First Grade College, Bidar, Karnataka 585402, 1 Department of Studies and Research in Chemistry, Gulbarga University, Gulbarga, Karnataka 585106, 2 Department of Post-Graduate Studies and Research in Chemistry, Sahyadri Science College, Kuvempu University, Shivamogga, Karnataka 577203, 3 Department of Chemistry Government Engineering College, Huvinahadagali, Karnataka 583219, India Accepted 10 November 2023 Revised 20 April 2023 Received 02 September 2022 Indian J Pharm Sci 2023;85(6):1660-1668 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms Fig. 1: Amiloride structure