Experimental design methodology for optimization and robustness determination in ion pair RP-HPLC method development: Application for the simultaneous determination of metformin hydrochloride, alogliptin benzoate and repaglinide in tablets

Publication date: June 2019Source: Microchemical Journal, Volume 147Author(s): Marianne A. Mahrouse, Nesrine T. LamieAbstractExperimental design methodology (DOE) surpasses traditional (one variable at a time) approach in improving the chromatographic separation performance with minimum effort and resources. Since combination therapy of oral hypoglycemic drugs was generally recommended by Clinical Practice Guidelines on hypoglycemic agent therapy for treatment of type 2 diabetes mellitus, DOE was adopted in development of an ion pair RP-HPLC method for the simultaneous determination of metformin hydrochloride (MET), alogliptin benzoate (ALO) and repaglinide (REP) in combined binary tablets. Screening using Plackett-Burman design followed by face centered composite design enabled the estimation of optimum settings that accomplish the most appropriate resolution and adequate peak shape within suitable run time. The designed models were statistically analyzed, graphically presented by surface plots and the relationships between coefficients of the derived polynomial equations were interpreted. The models fitted data adequately and could be used for response prediction. Chromatographic separation was achieved using Inertsil ODS column (250 mm, 4.6 mm, 5 μm), acetonitrile: phosphate buffer (0.01 M, adjusted to pH 2.5 with o-phosphoric acid): 0.3% sodium heptane sulfonate in water (60:20:20, v/v/v) as a mobile phase at flow rate 1 mL min−1. UV detection was carried...
Source: Microchemical Journal - Category: Chemistry Source Type: research