A Single-Run HPLC–MS Multiplex Assay for Therapeutic Drug Monitoring of Relevant First- and Second-Line Antibiotics in the Treatment of Drug-Resistant Tuberculosis
Niklas Köhler,
Hande Karaköse,
Hans-Peter Grobbel,
Doris Hillemann,
Sönke Andres,
Christina König,
Barbara Kalsdorf,
Thomas Theo Brehm,
Laura Böttcher,
Inna Friesen,
Harald Hoffmann,
Dražen Strelec,
Dagmar Schaub,
Charles A. Peloquin,
Stefan Schmiedel,
Laurent A. Decosterd,
Eva Choong,
Sebastian G. Wicha,
Rob E. Aarnoutse,
Christoph Lange,
Patricia M. Sánchez Carballo
Affiliations
Niklas Köhler
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Hande Karaköse
German Center for Infection Research (DZIF), Partner Site Borstel-Hamburg-Lübeck-Riems, 23845 Borstel, Germany
Hans-Peter Grobbel
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Doris Hillemann
National and World Health Organization Supranational Reference Laboratory for Mycobacteria, Research Center Borstel, 23845 Borstel, Germany
Sönke Andres
National and World Health Organization Supranational Reference Laboratory for Mycobacteria, Research Center Borstel, 23845 Borstel, Germany
Christina König
Department of Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
Barbara Kalsdorf
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Thomas Theo Brehm
Division of Infectious Diseases, I. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
Laura Böttcher
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Inna Friesen
National and World Health Organization Supranational Reference Laboratory for Mycobacteria, Research Center Borstel, 23845 Borstel, Germany
Harald Hoffmann
Institute of Microbiology and Laboratory Medicine, World Health Organization Supranational Reference Laboratory of TB, IML red GmbH, 82131 Gauting, Germany
Dražen Strelec
Department for Lung Diseases, Hospital for Lung Diseases and Tuberculosis, 42244 Klenovnik, Croatia
Dagmar Schaub
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Charles A. Peloquin
Infectious Disease Pharmacokinetics Laboratory, Emerging Pathogens Institute, University of Florida, Gainesville, FL 32610, USA
Stefan Schmiedel
Division of Infectious Diseases, I. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
Laurent A. Decosterd
Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, Switzerland
Eva Choong
Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, Switzerland
Sebastian G. Wicha
Institute of Pharmacy, University of Hamburg, 20246 Hamburg, Germany
Rob E. Aarnoutse
Department of Pharmacy, Radboud Institute for Medical Innovation, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands
Christoph Lange
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
Patricia M. Sánchez Carballo
Clinical Infectious Diseases, Research Center Borstel, Leibniz Lung Center, 23845 Borstel, Germany
The treatment of drug-resistant Mycobacterium tuberculosis relies on complex antibiotic therapy. Inadequate antibiotic exposure can lead to treatment failure, acquired drug resistance, and an increased risk of adverse events. Therapeutic drug monitoring (TDM) can be used to optimize the antibiotic exposure. Therefore, we aimed to develop a single-run multiplex assay using high-performance liquid chromatography–mass spectrometry (HPLC–MS) for TDM of patients with multidrug-resistant, pre-extensively drug-resistant and extensively drug-resistant tuberculosis. A target profile for sufficient performance, based on the intended clinical application, was established and the assay was developed accordingly. Antibiotics were analyzed on a zwitterionic hydrophilic interaction liquid chromatography column and a triple quadrupole mass spectrometer using stable isotope-labeled internal standards. The assay was sufficiently sensitive to monitor drug concentrations over five half-lives for rifampicin, rifabutin, levofloxacin, moxifloxacin, bedaquiline, linezolid, clofazimine, terizidone/cycloserine, ethambutol, delamanid, pyrazinamide, meropenem, prothionamide, and para-amino salicylic acid (PAS). Accuracy and precision were sufficient to support clinical decision making (≤±15% in clinical samples and ±20–25% in spiked samples, with 80% of future measured concentrations predicted to fall within ±40% of nominal concentrations). The method was applied in the TDM of two patients with complex drug-resistant tuberculosis. All relevant antibiotics from their regimens could be quantified and high-dose therapy was initiated, followed by microbiological conversion. In conclusion, we developed a multiplex assay that enables TDM of the relevant first- and second-line anti-tuberculosis medicines in a single run and was able to show its applicability in TDM of two drug-resistant tuberculosis patients.
