Startseite A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum
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A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum

  • Sankha S. Basu , Athena Petrides , Donald S. Mason und Petr Jarolim EMAIL logo
Veröffentlicht/Copyright: 21. November 2016
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Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 55 Heft 6

Abstract

Background:

Triazole antifungals are essential to the treatment and prophylaxis of fungal infections. Significant pharmacokinetic variability combined with a clinical need for faster turnaround times has increased demand for in-house therapeutic drug monitoring of these drugs, which is best performed using mass spectrometry-based platforms. However, technical and logistical obstacles to implementing these platforms in hospital laboratories have limited their widespread utilization. Here, we present the development and validation of a fast and simple ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to measure fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole in human serum suitable for incorporation into a hospital clinical laboratory.

Methods:

Serum samples (20 µL) were prepared using protein precipitation in the presence of deuterated internal standards. Chromatographic separation was accomplished using reversed phase UPLC and analysis was performed using positive-mode electrospray ionization and collision-induced dissociation MS.

Results:

Total analytical run time was 3 min. All analytes demonstrated linearity (r2>0.998) from 0.1 to 10 µg/mL (1–100 µg/mL for fluconazole), acceptable accuracy and precision (%DEV<15% and %CV<15% at all levels tested), suitable stability under relevant storage conditions, and correlated well with reference laboratory results.

Conclusions:

A simple and rapid UPLC-MS/MS method for monitoring multiple triazole antifungals was developed with a focus on the needs of hospital laboratories. The assay is suitable for clinical utilization and management of patients on these medications.

Keywords: liquid chromatography; mass spectrometry; method development; therapeutic drug monitoring; triazole antifungals
Corresponding author: Petr Jarolim, MD, PhD, Department of Pathology, Brigham and Women’s Hospital, 75 Francis Street, Boston, MA 02115, USA, Phone: +617-732-4725, Fax: +617-731-4872

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: P.J. has received research support from Abbott Laboratories, Amgen Inc., AstraZeneca LP, Beckman Coulter, Daiichi Sankyo, Inc., GlaxoSmithKline, Merck & Co., Inc., Roche Diagnostics Corporation, Takeda Global Research and Development Center, and Waters Technologies Corporation.

  3. Employment or leadership: D.S.M. is employed by and owns stock in Waters Corporation.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2016-0418) offers supplementary material, available to authorized users.

Received: 2016-5-16
Accepted: 2016-9-26
Published Online: 2016-11-21
Published in Print: 2017-6-1

©2017 Walter de Gruyter GmbH, Berlin/Boston

Artikel in diesem Heft

  1. Frontmatter
  2. Editorial
  3. Biotin interference on immunoassay methods: sporadic cases or hidden epidemic?
  4. Reviews
  5. False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences
  6. Vitamin K plasma levels determination in human health
  7. Mini Review
  8. Point-of-care testing INR: an overview
  9. Opinion Paper
  10. Could accreditation bodies facilitate the implementation of medical guidelines in laboratories?
  11. Genetics and Molecular Diagnostics
  12. QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions
  13. General Clinical Chemistry and Laboratory Medicine
  14. High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference
  15. Multicenter performance evaluation of a second generation cortisol assay
  16. A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum
  17. A microplate assay to measure classical and alternative complement activity
  18. Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2
  19. Retrospective evaluation of the clinical utility of serological biomarkers in Chinese patients with inflammatory bowel disease: 2-year clinical experience
  20. Infrared analysis of lipoproteins in the detection of alcohol biomarkers
  21. Coexistence of anti-β2-glycoprotein I domain I and anti-phosphatidylserine/prothrombin antibodies suggests strong thrombotic risk
  22. Antiphosphatidylserine/prothrombin antibodies as biomarkers to identify severe primary antiphospholipid syndrome
  23. Cardiovascular Diseases
  24. The impact of admission neutrophil-to-platelet ratio on in-hospital and long-term mortality in patients with infective endocarditis
  25. Letters to the Editor
  26. Biotin interference in immunoassays mimicking subclinical Graves’ disease and hyperestrogenism: a case series
  27. A simple method to detect biotin interference on immunoassays
  28. Proposed classification for a variant of Kounis syndrome
  29. Lyophilized hemoglobin E control material for the dichlorophenol-indophenol (DCIP) test
  30. Procalcitonin variation before and after 100-km ultramarathon
  31. Preliminary study in specific activity of molecular components in allergy: implications for diagnostics and relationship with disease severity
  32. A 2-min at 4500 g rather than a 15-min at 2200 g centrifugation does not impact the reliability of 10 critical coagulation assays
  33. Immunoassay interference caused by heterophilic antibodies interacting with biotin
  34. Red blood cell distribution width and mean platelet volume are potential prognostic indices for patients with primary biliary cirrhosis
  35. Neutrophil CD64 molecule expression can predict bloodstream infection in septic shock patients
https://doi.org/10.1515/cclm-2016-0418
Schlagwörter für diesen Artikel
liquid chromatography; mass spectrometry; method development; therapeutic drug monitoring; triazole antifungals

Artikel in diesem Heft

  1. Frontmatter
  2. Editorial
  3. Biotin interference on immunoassay methods: sporadic cases or hidden epidemic?
  4. Reviews
  5. False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences
  6. Vitamin K plasma levels determination in human health
  7. Mini Review
  8. Point-of-care testing INR: an overview
  9. Opinion Paper
  10. Could accreditation bodies facilitate the implementation of medical guidelines in laboratories?
  11. Genetics and Molecular Diagnostics
  12. QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions
  13. General Clinical Chemistry and Laboratory Medicine
  14. High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference
  15. Multicenter performance evaluation of a second generation cortisol assay
  16. A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum
  17. A microplate assay to measure classical and alternative complement activity
  18. Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2
  19. Retrospective evaluation of the clinical utility of serological biomarkers in Chinese patients with inflammatory bowel disease: 2-year clinical experience
  20. Infrared analysis of lipoproteins in the detection of alcohol biomarkers
  21. Coexistence of anti-β2-glycoprotein I domain I and anti-phosphatidylserine/prothrombin antibodies suggests strong thrombotic risk
  22. Antiphosphatidylserine/prothrombin antibodies as biomarkers to identify severe primary antiphospholipid syndrome
  23. Cardiovascular Diseases
  24. The impact of admission neutrophil-to-platelet ratio on in-hospital and long-term mortality in patients with infective endocarditis
  25. Letters to the Editor
  26. Biotin interference in immunoassays mimicking subclinical Graves’ disease and hyperestrogenism: a case series
  27. A simple method to detect biotin interference on immunoassays
  28. Proposed classification for a variant of Kounis syndrome
  29. Lyophilized hemoglobin E control material for the dichlorophenol-indophenol (DCIP) test
  30. Procalcitonin variation before and after 100-km ultramarathon
  31. Preliminary study in specific activity of molecular components in allergy: implications for diagnostics and relationship with disease severity
  32. A 2-min at 4500 g rather than a 15-min at 2200 g centrifugation does not impact the reliability of 10 critical coagulation assays
  33. Immunoassay interference caused by heterophilic antibodies interacting with biotin
  34. Red blood cell distribution width and mean platelet volume are potential prognostic indices for patients with primary biliary cirrhosis
  35. Neutrophil CD64 molecule expression can predict bloodstream infection in septic shock patients
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