Development, validation, and application of an HPLC method combined with an in vitro model for the determination of antibiotic binding to the haemoadsorber Cytosorb®

Kenda, Sara (avtor), Gubenšek, Jakob (avtor), Vovk, Tomaž (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: antibiotics, critically ill, CytoSorb®, gradient elution, haemoadsorption, hemoadsorption, in vitro model
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: FFA - Fakulteta za farmacijo ; MF - Medicinska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2026
• Št. strani: 14 str.
• Številčenje: Vol. 31, iss. 13, art. 2337
• PID: 20.500.12556/RUL-184399
• UDK: 61+615.33
• ISSN pri članku: 1420-3049
• DOI: 10.3390/molecules31132337
• COBISS.SI-ID: 283660803
• Datum objave v RUL: 06.07.2026

Gradivo je del revije

• Naslov: Molecules
• Skrajšan naslov: Molecules
• Založnik: MDPI
• ISSN: 1420-3049
• COBISS.SI-ID: 18462981

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: antibiotiki, kritično bolni, CytoSorb®, gradientna elucija, hemoadsorpcija, hemoadsorpcija, model in vitro, citokini, antibiotiki

Projekti

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P1-0189
• Naslov: Farmacevtska tehnologija: od dostavnih sistemov učinkovin do terapijskih izidov zdravil pri otrocih in starostnikih

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P3-0323
• Naslov: Ledvične bolezni in nadomestna zdravljenja