In vitro Study of Interactions of Sildenafil Citrate with Bovine Serum Albumin in Presence of Bisoprolol Fumarate and Metformin Hydrochloride by Fluorescence Spectrophotometry

Abstract

Aims: This paper describes the In vitro study of protein binding by sildenafil citrate (SC) in presence of bisoprololfumarate (BF) and metformin hydrochloride (MH). Study Design: Study was designed to assess In vitro of quenching of bovine serum albumin (BSA) by sildenafil citrate (SC) in presence of bisoprolol fumarate (BF) and metformin hydrochloride (MH) by fluorescence spectrophotometry. Place and Duration of Study: Drug Analysis and Research Laboratory, Centre for Advanced Research in Sciences, University of Dhaka, Dhaka-1000, Bangladesh between December 2013 and March 2014. Methodology: In the present work, the In vitro study of quenching of BSA by SC in presence of BF and MH have been studied by fluorescence emission spectroscopy under different conditions. At first, the BSA solution (20 µM) was prepared in phosphate buffer (pH =7.4) in eight test tubes and different amounts of sildenafil citrate was added to each BSA solution to obtain the final concentrations as 0, 20, 40, 80, 120, 160,240 and 320 × 10-6 molL-1, respectively. Then the fluorescence emission spectra of BSA-SC system were recorded for eight test tubes at two excitation wavelengths of BSA (λExmax= 280 nm and λExmax=293 nm) at 298 K and 308 K. Similarly, the fluorescence emission spectra of BSA-(SC+BF), BSA-(SC+MH) and BSA-(SC+BF+MH) systems were recorded at 280 nm and293 nm at 298 K and 308 K. Quenching constants were determined using the Stern-Volmer equation to provide a measure of the strength of quenching of BSA by SC in presence of BF and MH in all the systems. Results: The quenching of BSA by SC was increased in presence of BF and MH but remained close in presence of both BF and MH. Quenching constants were larger for the BSA-(SC+BF) system and ranked in the order as BSA-(SC+BF)>BSA-(SC+MH)>BSA-(SC+BF+MH)≈ BSA-SC at 280 nm at two different temperatures, respectively. But quenching at the excitation wavelength of 293 nm was ranked in order as BSA-(SC+BF) >BSA-(SC+MH) >BSA-(SC+BF+MH)>BSA-SC at 298 K and 308 K, respectively. Conclusion: It was found that BSA quenched by SC in presence of BF and MH, which indicated that the effectiveness of SC might be predominately influenced by these drugs.