Abstract
Although a marked decrease in mortality associated with bacterial infections is attributed to the discovery of antibiotics, antibiotic resistance has become a global health concern due to their misuse. A dynamic in vitro hollow-fiber system was used to study antibiotic resistance in Escherichia coli using ampicillin. An LC–MS/MS assay was validated for quantitative analysis of ampicillin in Luria–Bertani broth. The assay was linear from 0.10–50.00 μg/ml. The assay met acceptance criteria for inter- and intra-assay precisions and accuracies across three quality controls. Stability of ampicillin was confirmed at three different storage conditions. In vitro data were similar to simulated plasma PK data further confirming the appropriateness of the experimental design to quantify antibiotics and study occurrence of antimicrobial resistance in real-time.
Highlights
More and more bacteria are becoming increasingly resistant to the only treatment option: antibiotics
Research has been limited in bringing new antibiotics to the clinic and major effort is spent on revamping old antibiotics for curing bacterial infections
Recurring UTIs suggest that antibiotics are not very effective for all UTIs
Summary
Mass spectrometry optimization A LC–MS/MS method was developed with the purpose to have a sensitive and selective method suitable for quantification of AMPI from in vitro studies. Liquid chromatography & selectivity Chromatography conditions such as buffer strength, analytical column, and mobile composition and its flow rate were suitably altered to achieve better separation of the analytes from matrix components. Organic modifiers such as acetonitrile and methanol in combination with acidic modifiers such as formic acid were evaluated critically for their suitability to achieve better chromatographical resolution. Extraction recovery and matrix effect In this study, there was significant matrix effect (ion suppression) observed in LB broth for AMPI tested at LQC, MQC and HQC levels.
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