20.500.12556/RUL-184399 Development, validation, and application of an HPLC method combined with an in vitro model for the determination of antibiotic binding to the haemoadsorber Cytosorb® 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 (r$^2$2 > 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. antibiotics critically ill CytoSorb® gradient elution haemoadsorption hemoadsorption in vitro model antibiotiki kritično bolni CytoSorb® gradientna elucija hemoadsorpcija hemoadsorpcija model in vitro citokini antibiotiki true false true Angleški jezik Slovenski jezik Članek v reviji 2026-07-06 13:02:20 2026-07-06 13:02:26 2026-07-07 04:17:47 0000-00-00 00:00:00 2026 0 0 14 str. iss. 13, art. 2337 Vol. 31 Jul. 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 61+615.33 1420-3049 10.3390/molecules31132337 283660803 molecules-31-02337.pdf molecules-31-02337.pdf 1 0C8019A99046E1A9F154F4765EA1EC0D e42b6f3c92f3ad8cc56b9cb9d90e04b7fa83af32036b8e0f1dee5c4bcee05016 9f0a72ae-792a-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=237123 https://www.mdpi.com/1420-3049/31/13/2337 1 0f205c3f-792a-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=237122 Fakulteta za farmacijo Medicinska fakulteta 0 0 0