Novel method for determination of colistin sulfate in human plasma by high-performance liquid chromatography-tandem mass spectrometry and its clinical applications in critically ill patients

Abstract

Colistin is a last-resort antibiotic used for treating infections caused by carbapenem-resistant Gram-negative bacteria, particularly in critically patients, nevertheless its therapeutic window is narrow, and requires monitoring. A determination method suitable for clinical detection is conducive to ensure its efficacy and safety of patients with severe infection. We developed and validated a concise and accurate high-performance liquid chromatography-tandem mass spectrometry method for the determination of colistin A and B in human plasma. We used a Kinetex C18 column (50 mm × 2.1 mm, 2.6 μm) with acetonitrile (containing 0.1% formic acid) as the protein precipitant and water (containing 0.2% formic acid and 5 mmol/L ammonium formate) - acetonitrile (containing 0.2% formic acid) as the gradient elution. The calibration curves were linear over concentration ranges of 0.06–4.00 μg/mL (colistin A) and 0.1–7.0 μg/mL (colistin B). The precision, accuracy, matrix effect, extraction recovery, and stability were all validated. This method was applied to the therapeutic drug monitoring for 50 critically ill patients. The trough, peak, and average steady-state concentrations of these patients were 0.8 ± 0.4, 1.4 ± 0.5, and 1.0 ± 0.4 μg/mL, respectively. And the concentrations of colistin in human plasma were closely related to the patient's renal function.