Simultaneous Analysis of Antihyperlipidemic Drug Combinations by Green High‐Performance Liquid Chromatography Using Design of Experiments

ABSTRACT

Cardiovascular diseases are the leading cause of worldwide mortality, often supported by intricate dyslipidemic patterns of high levels of low‐density lipoprotein cholesterol and triacylglycerol. HMG‐CoA reductase inhibitors (statins) and ezetimibe can be used synergistically to treat these pathologies, as they target distinct metabolic pathways to reduce residual atherosclerotic risk. This research presents the design and multivariate optimization of a high‐performance liquid chromatographic system for the simultaneous determination of rosuvastatin, atorvastatin, simvastatin, and ezetimibe. The critical process parameters (i.e., the ratio of organic modifier and volumetric flow rate) were modeled using a three‐level central composite design to optimize chromatographic resolution and reduce overall analytical cycle time. The better separation was obtained on a Waters Sunfire C18 stationary phase (150 mm × 4.6 mm; 5 mm) using an isocratic elution system of 0.1% (v/v) aqueous acetic acid: acetonitrile (40:60, % v/v), at a flow rate of 1.4 mL/min, with spectrophotometric detection at 240 nm. The methodology was rigorously validated in line with ICH guidelines, demonstrating high selectivity, precision, accuracy, and linearity. Moreover, the method was critically evaluated with respect to green analytical chemistry parameters, ensuring the minimization of hazardous waste and solvent consumption. This proven framework, when successfully applied to binary marketed formulations, provides a robust, sustainable, and high‐throughput analytical methodology for routine quality control and for quantifying drug substances in multicomponent complex matrices.