Quantification of Hydroxychloroquine in Blood Using Turbulent Flow Liquid Chromatography-Tandem Mass Spectrometry (TFLC-MS/MS)

  • Allison B. Chambliss
  • Anna K. Füzéry
  • William A. ClarkeEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1383)


Hydroxychloroquine (HQ) is used routinely in the treatment of autoimmune disorders such as rheumatoid arthritis and lupus erythematosus. Issues such as marked pharmacokinetic variability and patient non-compliance make therapeutic drug monitoring of HQ a useful tool for management of patients taking this drug. Quantitative measurements of HQ may aid in identifying poor efficacy as well as provide reliable information to distinguish patient non-compliance from refractory disease. We describe a rapid 7-min assay for the accurate and precise measurement of HQ concentrations in 100 μL samples of human blood using turbulent flow liquid chromatography coupled to tandem mass spectrometry. HQ is isolated from EDTA whole blood after a simple extraction with its deuterated analog, hydroxychloroquine-d4, in 0.33 M perchloric acid. Samples are then centrifuged and injected onto the TFLC-MS/MS system. Quantification is performed using a nine-point calibration curve that is linear over a wide range (15.7–4000 ng/mL) with precisions of <5 %.

Key words

Hydroxychloroquine Therapeutic drug monitoring Turbulent flow liquid chromatography Tandem mass spectrometry Blood Quantification 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Allison B. Chambliss
    • 1
  • Anna K. Füzéry
    • 1
  • William A. Clarke
    • 1
    Email author
  1. 1.Department of Pathology - Clinical ChemistryJohns Hopkins University School of MedicineBaltimoreUSA

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