Development and application of an LC-MS method with high resolution accurate masses detection (HRAM) with ExactivePlus mass spectrometer and online sample preparation for antibiotics analysis
Valérie Thibert1,*, François Coudoré2, Alban Lemonnier2
1Chromatography and Mass Spectrometry Division, Thermo Fisher Scientific France, Courtaboeuf cedex, 2Laboratoire de microbiologie clinique et de dosage des anti-infectieux, Groupe Hospitalier Paris Saint Joseph, Paris, France
Introduction: There is an increasing interest in quantifying antibiotics in plasma. Liquid chromatography coupled to mass spectrometry seems like a good option for this analysis considering the differences in size and polarity of antibiotics.
Objectives: The goal of this work was to develop and apply an LC-MS method for the analysis of 24 β-lactam antibiotics.
Methods: The optimized method consisted in the TurboFlow extraction of the antibiotics from human plasma samples with a Cyclone MAX and a Cyclone MCX2 columns, coupled to a Hypersil Gold Aq column for separation. The detection was performed with an Exactive Plus mass spectrometer with electrospray ionization in positive and negative FullScan mode with a resolution of 35,000 (FWHM at m/z 200).
Results: The developed method showed global recoveries, considering matrix effects, superior to 70% for 22 of the molecules and between 50% and 60% for two molecules. The method was validated for each molecule in terms of sensitivity, linearity, precision and accuracy.
Conclusion: The developed method is rapid and accurate. The use of TurboFlow extraction minimizes importantly the sample pre-treatment, while the use of HRAM leads to a simple and very specific detection.
Keywords: Blood, Method