Box-Behnken Assisted Validation And Optimization Of An Rp-Uplc Method For Simultaneous Determination Of Lobeglitazone Sulfate And Glimepiride

A simple, precise, and accurate reverse-phase high-performance liquid chromatography (RP-UPLC) method was developed for the simultaneous estimation of lobeglitazone sulfate and glimepiride in bulk and tablet dosage forms. The method involved selecting various chromatographic parameters. Specifically, a new method was developed using a UPLC HSS C18 column with diameter of 1.8 µm, 2.1 mm X 100 mm. The mobile phase consisted of 0.1N Potassium dihydrogen Ortho phosphate (pH 4.8)(61 v/v) and Acetonitrile (39 v/v), running as an isocratic elution. The flow rate was set at 0.3 mL/min, and UV detection was performed at 248 nm. The injection volume was 1 µL, and the total runtime was 3 minutes. The recoveries for lobeglitazone sulfate were found to be in the range of 99.7%, while those for glimepiride were in the range of 100.2%. The method’s accuracy is evident from these results. The inter-day and intra-day precision of the new method were both below the maximum allowable limit (RSD% < 2.0), as per International Council on Harmonisation (Q2 R1) guidelines. The method exhibited a linear response, with correlation coefficient (r2) values of 0.999 for lobeglitazone sulfate and 0.999 for glimepiride. The validation parameters, such as accuracy, precision, linearity, specificity, stability in the analytical solution and robustness, met the acceptance criteria. Hence, this method can be effectively used for the simultaneous estimation of lobeglitazone sulfate and glimepiride in both bulk and pharmaceutical dosage forms. Method optimization was achieved using Box-Behnken design software ,in which three key variables were examined, namely, the flow rate, the composition of mobile phase and the temperature on responses like retention time (Y1-2), resolution factor (Y3), theoretical plates (Y4-5), and tailing factor (Y6-7), a mathematical model was created.We observed that slight alteration in the mobile phase and flow rate influenced the outcome. This method is potentially useful for the analysis of commercial formulations and laboratory preparations. Results: Design expert for suitable chromatographic conditions to choose an appropriate solvents system or mobile phase  a Qbd tested a number of solvents , including Potassium dihydrogen phosphate and acetonitrile. Finally in isocratic mode of separation , finally it is optimized using a UPLC HSS C18 column with diameter of 1.8 µm, 2.1 mm X 100 mm. injection volume of 1.0 mL  and flow rate of 0.3ml/min design expert 11 software was used to examine the data collected. The programm recommended a quadratic model. Variables like Y1,Y2,Y3,Y4,Y5,Y6,Y7. For each dependent variables, ANOVA was used to create mathematical models which were when translated into 3D surface graph and equations . The mobile phase composed of 61.v/v  0.1N Potassium dihydrogen phosphate(PH 4.8)  and acetonitrile 39.v/v was choosen due to its polar composition for the efficient separation of our analytes lobeglitazone sulfate and glimepride. Analytical quality by design (QbD) with UPLC method for the estimation of lobeglitazone and glimepride combined dosage forms . Target profile was retention time, plate count, resolution and tailing factor . Mobile phase ratio, flow rate , length  of column and temperature were identified as critical quality attributes. Quality by Design was applied for two level factorial design by Design expert 11 software was used. The quality by Design approach was successful. Different method validation parameters like linearity correlation should not less than 0.999. the precisison for percentage recovery is 99.7./. and 100.2./. LOD and LOQ for drug lobeglitazone 0.03 and 0.10 µg/mL   and for glimepride is 0.06 and 0.18 µg/mL .Degradation studies have done for lobeglitazone and glimepride  the acid degradation values are 1.112 and 1.444 , alkaline degradation values are 1.114 and 1.436, peroxide degradation 1.102 and 1.435 , thermal degradation values are 1.134 and 1.434 , photolytic degradation values are 1.101 and 1.431 neutral hydrolysis  values 1.107 and 1.439 the developed method was validated according to ICH guidelines. Conclusions: A fast, accurate, specific, and stability-indicating RP-UPLC method has been developed for the quantitative detection of Lobeglitazone sulphate and Glimepiride in pharmaceutical dosage forms using the AQbD-based methodology.