A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring

Cystic fibrosis (CF) is a life-threatening disorder caused by mutations in the CFTR gene, leading to defective chloride ion transport and thickened mucus in the respiratory and gastrointestinal systems. CFTR modulators, including ivacaftor, lumacaftor, tezacaftor, and elexacaftor, have improved pati...

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Main Authors: Valentina D’Atri, Fabrizio Corrado, François Versace, Susana Alves Saldanha, Thomas Mercier, Monia Guidi, Paul Thoueille, Sylvain Blanchon, Angela Koutsokera, Michael Vogeser, Catia Marzolini, François Girardin, Georgia Mitropoulou, Zisis Balmpouzis, Isabelle Rochat, Alain Sauty, Laurent Arthur Decosterd, Eva Choong
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/9/1866
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author Valentina D’Atri
Fabrizio Corrado
François Versace
Susana Alves Saldanha
Thomas Mercier
Monia Guidi
Paul Thoueille
Sylvain Blanchon
Angela Koutsokera
Michael Vogeser
Catia Marzolini
François Girardin
Georgia Mitropoulou
Zisis Balmpouzis
Isabelle Rochat
Alain Sauty
Laurent Arthur Decosterd
Eva Choong
author_facet Valentina D’Atri
Fabrizio Corrado
François Versace
Susana Alves Saldanha
Thomas Mercier
Monia Guidi
Paul Thoueille
Sylvain Blanchon
Angela Koutsokera
Michael Vogeser
Catia Marzolini
François Girardin
Georgia Mitropoulou
Zisis Balmpouzis
Isabelle Rochat
Alain Sauty
Laurent Arthur Decosterd
Eva Choong
author_sort Valentina D’Atri
collection DOAJ
description Cystic fibrosis (CF) is a life-threatening disorder caused by mutations in the CFTR gene, leading to defective chloride ion transport and thickened mucus in the respiratory and gastrointestinal systems. CFTR modulators, including ivacaftor, lumacaftor, tezacaftor, and elexacaftor, have improved patient outcomes, but interindividual pharmacokinetic variability and potential drug–drug interactions require therapeutic drug monitoring (TDM) for optimal efficacy and safety. In this context, a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantification of CFTR modulators and their major active metabolites in human plasma to support pharmacokinetic studies and routine TDM. The multiplex LC-MS/MS assay was established using plasma protein precipitation, followed by chromatographic separation on an Xselect HSS T3 (Waters<sup>®</sup>) column and positive electrospray ionization mode detection. The method was validated based on FDA and EMA guidelines for specificity, linearity, accuracy (89.8–107.8%), repeatability (1.1–8.1%), intermediate fidelity (1.3–10.9%), matrix effects, and stability, demonstrating a robust performance with excellent precision and accuracy. International interlaboratory comparisons confirmed the reliability of the assay. The developed method can be applied for the clinical monitoring of caftors’ plasma concentrations and preliminary data suggest that it can also be applied to alternative matrices, such as breast milk. This method will serve to characterize caftors’ pharmacokinetic variability and monitor drug–drug interactions to further refine personalized dosing strategies and enhance precision medicine treatments for patients with CF.
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spelling doaj-art-45928534da5e41dfa358f10acef4a3c02025-08-20T02:58:47ZengMDPI AGMolecules1420-30492025-04-01309186610.3390/molecules30091866A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic MonitoringValentina D’Atri0Fabrizio Corrado1François Versace2Susana Alves Saldanha3Thomas Mercier4Monia Guidi5Paul Thoueille6Sylvain Blanchon7Angela Koutsokera8Michael Vogeser9Catia Marzolini10François Girardin11Georgia Mitropoulou12Zisis Balmpouzis13Isabelle Rochat14Alain Sauty15Laurent Arthur Decosterd16Eva Choong17Service and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandPediatric Pneumology and Cystic Fibrosis Unit, Division of Pediatrics, Department Woman-Mother-Child, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandAdult Cystic Fibrosis and CFTR-Related Disorders Center, Division of Pulmonology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandInstitute of Laboratory Medicine, LMU University Hospital, 81377 Munich, GermanyService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandAdult Cystic Fibrosis and CFTR-Related Disorders Center, Division of Pulmonology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandAdult Cystic Fibrosis and CFTR-Related Disorders Center, Division of Pulmonology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandPediatric Pneumology and Cystic Fibrosis Unit, Division of Pediatrics, Department Woman-Mother-Child, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandAdult Cystic Fibrosis and CFTR-Related Disorders Center, Division of Pulmonology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandService and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, Lausanne University Hospital and University of Lausanne, 1011 Lausanne, SwitzerlandCystic fibrosis (CF) is a life-threatening disorder caused by mutations in the CFTR gene, leading to defective chloride ion transport and thickened mucus in the respiratory and gastrointestinal systems. CFTR modulators, including ivacaftor, lumacaftor, tezacaftor, and elexacaftor, have improved patient outcomes, but interindividual pharmacokinetic variability and potential drug–drug interactions require therapeutic drug monitoring (TDM) for optimal efficacy and safety. In this context, a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantification of CFTR modulators and their major active metabolites in human plasma to support pharmacokinetic studies and routine TDM. The multiplex LC-MS/MS assay was established using plasma protein precipitation, followed by chromatographic separation on an Xselect HSS T3 (Waters<sup>®</sup>) column and positive electrospray ionization mode detection. The method was validated based on FDA and EMA guidelines for specificity, linearity, accuracy (89.8–107.8%), repeatability (1.1–8.1%), intermediate fidelity (1.3–10.9%), matrix effects, and stability, demonstrating a robust performance with excellent precision and accuracy. International interlaboratory comparisons confirmed the reliability of the assay. The developed method can be applied for the clinical monitoring of caftors’ plasma concentrations and preliminary data suggest that it can also be applied to alternative matrices, such as breast milk. This method will serve to characterize caftors’ pharmacokinetic variability and monitor drug–drug interactions to further refine personalized dosing strategies and enhance precision medicine treatments for patients with CF.https://www.mdpi.com/1420-3049/30/9/1866cystic fibrosisCFTR modulatorstherapeutic drug monitoringLC-MS/MSpharmacokineticsprecision medicine
spellingShingle Valentina D’Atri
Fabrizio Corrado
François Versace
Susana Alves Saldanha
Thomas Mercier
Monia Guidi
Paul Thoueille
Sylvain Blanchon
Angela Koutsokera
Michael Vogeser
Catia Marzolini
François Girardin
Georgia Mitropoulou
Zisis Balmpouzis
Isabelle Rochat
Alain Sauty
Laurent Arthur Decosterd
Eva Choong
A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
Molecules
cystic fibrosis
CFTR modulators
therapeutic drug monitoring
LC-MS/MS
pharmacokinetics
precision medicine
title A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
title_full A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
title_fullStr A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
title_full_unstemmed A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
title_short A Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Cystic Fibrosis Drugs (Caftors) in Plasma and Its Application for Therapeutic Monitoring
title_sort liquid chromatography tandem mass spectrometry method for the quantification of cystic fibrosis drugs caftors in plasma and its application for therapeutic monitoring
topic cystic fibrosis
CFTR modulators
therapeutic drug monitoring
LC-MS/MS
pharmacokinetics
precision medicine
url https://www.mdpi.com/1420-3049/30/9/1866
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