Simultaneous Determination of Anti-diabetic Drugs in Their Combined Dosage Forms Using HPLC: An Experimental Design Approach

Abstract

A liquid chromatographic method has been established for the separation of five presently existing anti-diabetic drugs-metformin, pioglitazone, glibenclamide, glimepiride and repaglinide using design of experiments methodology. The main objective of this method is to identify the suitable conditions for the adequate separation of these five drugs with minimal analysis time. Experiment design is one of the Quality by design approaches, which incorporates multifaceted combinations of independent variables and their interactions during the optimization of analytical methods for the determination of drugs. Phenomenex Luna C8-type column (250 × 4.6 mm; 5 μm particle size) was employed and 0.3% acetic acid in water: acetonitrile mixture was used as mobile phase (0.829 mL min-1 as flow rate) for the separation of analytes (Detection wavelength-230 nm). The chromatographic efficiency was investigated for the combined effects of each of mobile phase composition, buffer strength, flow rate at three levels using Box-Behnken design which is one of experimental design. The optimized method was validated using the above statistical approach and applied for the assay of pharmaceutical formulations.