Reversed-phase high-performance liquid chromatography analytical method development and validation for cefepime and sulbactam injection formulation assay: A quality by design approach

A trustworthy high-performance liquid chromatography approach for the simultaneous measurement of cefepime and sulbactam was created using quality by design. Finding reliable chromatographic settings for quality peaks and suitable component separation within a manageable run time was the key objective. Critical quality attributes for risk measurement were evaluated. To create an analytical design space and create a control strategy, the quantitative link between critical material attributes and critical quality attributes was built. Acetonitrile volume, trimethylamine volume, and flow rate were the three independent parameters considered in the mathematical models. Acetonitrile:high-performance liquid chromatography water:triethylamine and pH 6.0, adjusted with orthophosphoric acid (50.0:50:0.15, v/v/v) as the mobile phase at 1.1 ml/min, were the ideal and anticipated conditions. We obtained appropriate baseline separation of both medications in about 10 min using these ideal conditions. The main determinant of the perturbation plot responses was found to be the quantity of acetonitrile. At all three levels, the percent recoveries for both medications ranged from 99% to 99.68%. Relative standard deviations were less than 2% for both drugs' intermediate accuracy and repeatability. A straightforward, precise, and repeatable high-performance liquid chromatography method for regular quality control testing was created as a consequence.

Automated summary

A trustworthy high-performance liquid chromatography approach was developed using quality by design for simultaneous measurement of cefepime and sulbactam. Chromatographic settings that ensure quality peaks and component separation within a manageable run time were determined. Critical quality attributes were evaluated to establish an analytical design space and control strategy. The independent parameters in the mathematical models were acetonitrile volume, trimethylamine volume, and flow rate. The ideal conditions were found to be the mobile phase of acetonitrile:high-performance liquid chromatography water:triethylamine at a ratio of 50.0:50:0.15 (v/v/v), pH 6.0 adjusted with orthophosphoric acid, and a flow rate of 1.1 ml/min. The developed method provided appropriate baseline separation of both drugs within approximately 10 minutes with high percent recoveries and low relative standard deviations.