Chemometric Assisted HPLC Method for the Simultaneous Estimation of Aspirin, Atorvastatin and Clopidogrel in Biological Matrix

Aim: A new efficient simple and robust liquid chromatographic method has been developed and validated for the simultaneous estimation of aspirin, atrovastatin and clopidogrel in human plasma sample.

Study Design: The investigation was focused on studying the influence of organic phase modifier, pH and flow rate, which modify the separation of chromatographic behavior of these compounds.

Methodology: To optimize the chromatographic factors that had a significant effect on separation attributes central composite design was applied. The global optimization of the chromatographic responses such as capacity factor, resolution and analysis time optimized derringer’s desirability function was employed.

Results: The optimized chromatographic condition was Acetonitrile: Methanol: 0.1% Triethylamine (pH 3.0 adjusted with Ortho phosphoric acid) (53.55: 05: 41.45 % v/v), 1.14 ml/min flow rate and measurement was performed PDA at 220 nm. The spiked plasma sample was extracted by a simple protein precipitation method and then the sample was extracted with cold methanol and solvents was evaporated by the dry air method.

Conclusion: This method can be useful for the computable determination of aspirin, atrovastatin and clopidogrel in human plasma suitable for bioequivalence and pharmacokinetic studies in healthy human volunteers.