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Updated: Jul 16, 2026
Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
Published on: August 30, 2018
Development, Validation, and Application of an HPLC Method Combined with an In Vitro Model for the Determination of
Sara Kenda1,2, Jakob Gubenšek3,4, Tomaž Vovk1
1Faculty of Pharmacy, University of Ljubljana, Askerceva 7, 1000 Ljubljana, Slovenia.
Supportive therapy with haemoadsorption is gaining popularity in critically ill patients, with the aim of reducing overinflammation triggered by the cytokine storm. The haemoadsorbers used are not specific for cytokines and also bind antibiotics. The aim of this study was to develop and validate a simple analytical method for the simultaneous determination of selected antibiotics and to develop an in vitro model for the quantification of their binding to the CytoSorb® haemoadsorber under conditions simulating sepsis. Imipenem (IMI), amoxicillin (AMO), cefepime (CEF), meropenem (MERO), vancomycin (VAN) and piperacillin (PIP) were measured in bovine plasma via precipitation with acetonitrile and liquid-liquid extraction with dichloromethane. The aqueous phase was collected and analysed using a C18 HPLC system under gradient conditions, with modulation of organic solvent content and mobile phase pH, and detection performed using a UV/Vis detector. The method was linear (r2 > 0.982) across investigated analytical ranges (1.0-100.0 µg/mL for AMO and VAN, 1.0-75.0 µg/mL for CEF, MERO and PIP and 2.5-100.0 µg/mL for IMI). Intra- and inter-day precision did not exceed 14% and accuracy ranged from 85.8% to 108.5%. Using the in vitro model, we showed that CytoSorb® significantly removed VAN and PIP, but not MERO. Further clinical studies are needed to establish the clinical significance of these findings and their impact on antibiotic exposure.
Supportive therapy with haemoadsorption is gaining popularity in critically ill patients, with the aim of reducing overinflammation triggered by the cytokine storm. The haemoadsorbers used are not specific for cytokines and also bind antibiotics. The aim of this study was to develop and validate a simple analytical method for the simultaneous determination of selected antibiotics and to develop an in vitro model for the quantification of their binding to the CytoSorb® haemoadsorber under conditions simulating sepsis. Imipenem (IMI), amoxicillin (AMO), cefepime (CEF), meropenem (MERO), vancomycin (VAN) and piperacillin (PIP) were measured in bovine plasma via precipitation with acetonitrile and liquid-liquid extraction with dichloromethane. The aqueous phase was collected and analysed using a C18 HPLC system under gradient conditions, with modulation of organic solvent content and mobile phase pH, and detection performed using a UV/Vis detector. The method was linear (r2 > 0.982) across investigated analytical ranges (1.0-100.0 µg/mL for AMO and VAN, 1.0-75.0 µg/mL for CEF, MERO and PIP and 2.5-100.0 µg/mL for IMI). Intra- and inter-day precision did not exceed 14% and accuracy ranged from 85.8% to 108.5%. Using the in vitro model, we showed that CytoSorb® significantly removed VAN and PIP, but not MERO. Further clinical studies are needed to establish the clinical significance of these findings and their impact on antibiotic exposure.
