Box-Behnken Design Assisted Optimization and Standardization of Chromatographic Methodology for Quality Assessment of Metformin: Analytical Quality by Design Avenue

Abstract:

Background: Metformin (MET) is an oral antidiabetic agent falls chemically under the category of biguanides and it is effectively utilized in the management of type 2 diabetes mellitus. It is marketed in the tablet dosage form and hence quality control and assessment of MET is very much essential and important. Objectives: The objectives of present research investigation are to implement the Box-Behnken Design (BBD) in the optimization and validation of Reverse Phase-High Performance Liquid Chromatographic (RP-HPLC) method for the quality assessment of MET and also to systematically apply the Analytical Quality by Design (AQbD) Approach. Materials and Methods: In methodology BBD was employed to identify and optimize the critical method aspects for augmenting the performance of proposed methodology. The optimum chromatographic elution was performed on Agilent C₁₈ (5μm, 250 × 4.6 mm i.d.) column employing methanol and 0.1% ortho phosphoric acid in water with pH 2.8 in the proportions of 4:96 v/v as solvent system. The elution was carried out at 0.7 mL/min flow of mobile phase and identification at 231 nm using UV detector. The BBD driven optimized method was standardized as per guidelines of International Council for Harmonisation Q2 (R₁) in terms of validating method parameters such as specificity, linearity, detection and quantitation limit, precision, robustness, ruggedness and accuracy. Results: The method was linear in the concentration of 5-25μg/ml with R² = 0.999. The detection and quantitation limit were obtained at concentration of 0.30 and 0.93 μg /ml. The values of precision, robustness, and ruggedness parameters were found to be well within the required limit of acceptance with deviation of relative standard < 2%. The accuracy of MET was observed 99.22% to 100.25% at three different recovery experiments. At the end the proposed design of experiment (DoE) oriented methodology successfully applied for quantification of MET. Conclusion: The BBD and AQbD approaches are highly useful for the quality assessment of MET in bulk and its marketed tablet dosage forms.

Key words: Analytical Quality by Design, Box–Behnken Design, Chromatography, Metformin, Standardization,