The application of experimental design and green analytical chemistry in HPLC strategy for the determination of five drugs used in antimicrobial combination therapy

Abstract

Antimicrobial resistance is a significant global issue that has increased mortality rates over the past 100 years. One of the strategies followed to overcome this problem is to use antimicrobial combination therapy with different action modes that can synergize for successful treatment in emergent situations. Ciprofloxacin, Tinidazole, Dexamethasone Sodium Phosphate, Diloxanide Furoate, Metronidazole, and their different combinations are commonly used as effective combination therapies in critical cases. Hence, it is essential to establish an appropriate technique for their simultaneous analysis. The application of green analytical chemistry concepts was also an important issue that should be considered in the development of analytical methods. Hence, a new green micellar organic solvent-free HPLC methodology has been devised to analyze the specified five drugs simultaneously in both their authentic forms and combined pharmaceutical formulations. Response surface methodology was used to investigate the influence of different variable factors on chromatographic performance. Central composite design was utilized for response optimization. A Symmetry C18 column (3.5 µm, 4.6 × 75 mm), an isocratic mixed–micellar mobile phase composed of (70.76 mM of sodium dodecyl sulphate and 21.38 mM of Brij-35, 2.76 pH, and 40 ± 2 °C at a flow rate of 1 mL/min, were the ideal separation circumstances for analytesʹ separation at 260 nm as a chosen detection wavelength. The described approach was satisfactorily utilized todetermine various pharmaceutical formulations of the specified medications in combination. ICH guidelines were followed during the method's validation. Analytical Greenness Measure (AGREE) and Green Analytical Procedure Index (GAPI) fundamentals were followed to judge the developed methodology's greenness.