development and validation of a rapid lc-ms/ms method for simultaneous quantification of atorvastatin, candesartan, and clopidogrel in human plasma and solutions for pharmaceutical and clinical applications

This study presents the development and validation of a novel, robust liquid chromatography–tandem mass spectrometry (lc–ms/ms) method for the simultaneous quantification of atorvastatin, candesartan, and clopidogrel in solution and human plasma. the method complies with european medicines agency (ema) guidelines and utilizes an isocratic mobile phase composed of 0.1% acetic acid and methanol (2:8, v/v), adjusted to ph with 0.02% triethylamine. separation was achieved on a c18 column with retention times of 2.37 minutes for plasma and 1.5 minutes for solution. the method demonstrated high sensitivity with a lower limit of quantification (lloq) of 0.005 µg/ml and excellent linearity across the concentration range of 0.005–0.5 µg/ml (r² > 0.999 for all analytes). precision and accuracy were within ema-accepted limits, with accuracy ranging from 91.9% to 106.2% and coefficient of variation (cv) values between 1.2% and 14.4%. the method showed no interference from endogenous matrix components and met selectivity criteria. compared to existing approaches, it offers reduced solvent consumption, faster run times, and lower sample volume requirements. additionally, the method shows promise for therapeutic drug monitoring in alternative biological matrices, such as saliva. these findings establish the lc–ms/ms method as a highly efficient and reliable tool for cardiovascular combination therapy research, with applications in clinical and pharmacokinetic studies.