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Updated: Jun 20, 2025

Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
Published on: August 30, 2018
Free serum concentrations of antibiotics determined by ultrafiltration: extensive evaluation of experimental
Constantin Lier1, Alexander Dejaco2, Alexander Kratzer3
1Institute of Pharmacy, University of Regensburg, Regensburg, Germany.
Abstract:
Aim: To assess the impact of experimental conditions on free serum concentrations as determined by ultrafiltration and HPLC-DAD analysis in a wide range of antibiotics.Materials & methods: Relative centrifugation force (RCF), temperature, pH and buffer were varied and the results compared with the standard protocol (phosphate buffer pH 7.4, 37°C, 1000 × g).Results: Generally, at 10,000 × g the unbound fraction (fu) decreased with increasing molecular weight, and was lower at 22°C. In unbuffered serum, the fu of flucloxacillin or valproic acid was increased, that of basic or amphoteric drugs considerably decreased. Comparable results were obtained using phosphate or HEPES buffer except for drugs which form metal chelate complexes.Conclusion: Maintaining a physiological pH is more important than strictly maintaining body temperature.
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