Development of an Analytical Method for Ligand Quantification in Pharmaceutical Forms Using Visible Range Molecular Absorption Spectroscopy

Abstract

    The research includes for the assessment of ligands as raw materials and in pharmaceutical preparations, the research entails the development of a sensitive, cost-effective, and specific spectroscopic analytical approach. The suggested technique uses a nucleophilic substitution mechanism to create an orange-red complex that exhibits distinctive absorbance at 450 nm. The optimal experimental conditions for the reaction were studied, with the best conditions being the use of 1.4 ml of reagent at a concentration of 2.2% w/v, in the presence of methanol as a solvent, and heating to 90°C in a water bath for 22 minutes. The stability of the product and the ratio of the reactants involved in its formation were also studied, according to the Job method and the molar ratio method. The results showed that the resulting complex was stable for up to two hours, and that the reaction ratio was one to one for each reagent. The analytical method was evaluated according to ICH rules under optimum reaction conditions. Lambert-Beer's law was applied within concentrations (30-110 M) with a correlation coefficient of R=0.99 and recovery values ranging from 99.39-99.59%. The method was successfully used to determine the ligand in tablets, as no obstruction was observed from the excipients included in the composition of these preparations or metformin in the participating pharmaceutical forms. It was found that there were no statistically significant differences when comparing the results with the results of the assay by HPLC and UV-vis absorption methods in terms of t-test and F-test. The finding results of the docked compound Gliclazide with binding affinity – 7.7 kcal/mol, the results show us the compound's ability to inhibit the enzyme UNC, which promises to inhibit and treat inflammation in the udder of cows, thus achieving the highest possible levels of milk production.