Development and validation of a quantitative HPLC/MS/MS method for the determination of piperacillin in blood plasma

Abstract

Introduction: Reduced mortality in patients with sepsis taking piperacillin is possible when they receive a long-term infusion, which improves the effect of antimicrobials. However, such piperacillin therapy requires therapeutic drug monitoring, the use of the latest analytical equipment and developed methods for the quantitative determination of piperacillin. Materials and Methods: Dry samples of the appropriate certified piperacillin standards were used to prepare stock and standard solutions of piperacillin. Separation of the components was performed using an Agilent 1260 HPLC system with a binary pump and a temperature controlled autosampler. Analyses were detected using a Sciex QTRAP 5500 hybrid mass spectrometric system. Validation of the developed method was carried out in accordance with the rules for conducting bioequivalence studies of drugs within the framework of the Eurasian Economic Union; 2016, in Astana. Results and Discussion: Piperacillin ions-”precursors” corresponded to particles m/z 518.2. The most intense ions-”products” registered during the fragmentation of protonated molecules in the collision cell were particles m/z 143.1 and m/z 115.0. During the validation of the developed method, the main validation parameters were established: linearity, accuracy, accuracy, and sensitivity (lower limit of quantitation). Conclusion: The validated analytical range of the method was 0.5–100 µg/mL in plasma. The resulting analytical range makes it possible to apply the developed method for conducting the analytical part of studies of the pharmacokinetics of piperacillin.

Keywords

• hplc/ms
• validation
• quantitation
• piperacillin
• bioanalytics