Startseite Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
Artikel
Lizenziert
Nicht lizenziert Erfordert eine Authentifizierung

Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)

  • Kai Bruns , Rene Mönnikes und Karl J. Lackner EMAIL logo
Veröffentlicht/Copyright: 7. Dezember 2015
Veröffentlichen auch Sie bei De Gruyter Brill
Manuskript einreichen Informationen für Autor*innen
Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 54 Heft 7

Abstract

Background: Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) utilizing triple-quadrupole instruments has been widely used for quantification of endogenous compounds, drugs or metabolites in clinical laboratories. In contrast, high-resolution mass spectrometry (HRMS) is typically used for compound identification due to its limited dynamic range. Recently HRMS instruments with enhanced linear dynamic range have become available. The aim of this study was to evaluate HRMS for fast quantitative applications in a clinical laboratory.

Methods: A high throughput UPLC-TOF-MS assay for simultaneous quantification of cyclosporin A, tacrolimus, sirolimus and everolimus was developed. All immunosuppressants were analyzed as sodium adducts in TOF-only mode using an Agilent 6540 Q-TOF system. Extracted ion chromatograms of analytes and internal standards were created from full-scan data. The assay was evaluated and compared to an established LC-MS/MS assay according to CLSI recommendations.

Results: The novel HRMS assay has a total run time of 3 min. The assay is linear in a clinical relevant concentration range for all four immunosupressants. Method correlations vs. established LC-MS/MS assay were between R2=0.99 and R2=0.97. Total coefficients of variation (CVT) ranges were 4.5%–6.4% (tacrolimus), 7.4%–8.0% (sirolimus), 8.0%–8.8% (everolimus) and 6.1%–7.4% (cyclosporine A) for three relevant concentration levels each.

Conclusions: High resolution TOF-MS and LC-MS/MS show equivalent quantitative performance for monitoring of cyclosporin A, tacrolimus, sirolimus and everolimus. HRMS has the potential to replace conventional LC-MS/MS in clinical laboratories because it simplifies assay development (no optimization of fragmentations and product ions necessary) and its full-scan data can provide additional information.

Keywords: high resolution mass spectrometry; immunosuppressants; LC-MS/MS; method validation; TOF-MS
Corresponding author: Karl J. Lackner, MD, Institute of Clinical Chemistry and Laboratory Medicine, University Medical Centre Mainz, Langenbeckstr. 1, 55131 Mainz, Germany, Phone: +49 6131 177190, Fax: +49 6131 176627, E-mail:

References

1. Adaway JE, Keevil BG, Owen LJ. Liquid chromatography tandem mass spectrometry in the clinical laboratory. Ann Clin Biochem 2015;52:18–38.10.1177/0004563214557678Suche in Google Scholar PubMed

2. Ombrone D, Giocaliere E, Forni G, Malvagia S, La Marca G. Expanded newborn screening by mass spectrometry: new tests. future perspectives. Mass Spectrom Rev, [Epub ahead of print], doi:10.1002/mas.21463.Suche in Google Scholar PubMed

3. Eckart K, Röhrich J, Breitmeier D, Ferner M, Laufenberg-Feldmann R, Urban R. Development of a new multi-analyte assay for the simultaneous detection of opioids in serum and other body fluids using liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2015;1001:1–8.10.1016/j.jchromb.2015.06.028Suche in Google Scholar PubMed

4. Müller MJ, Volmer DA. Mass spectrometric profiling of vitamin D metabolites beyond 25-hydroxyvitamin D. Clin Chem 2015;61:1033–48.10.1373/clinchem.2015.241430Suche in Google Scholar PubMed

5. Keefe CC, Goldman MM, Zhang K, Clarke N, Reitz RE, Welt CK. Simultaneous measurement of thirteen steroid hormones in women with polycystic ovary syndrome and control women using liquid chromatography-tandem mass spectrometry. PLoS ONE 2014;9:e93805.10.1371/journal.pone.0093805Suche in Google Scholar PubMed PubMed Central

6. Krüger R, Bruns K, Grünhage S, Rossmann H, Reinke J, Beck M, et al. Determination of globotriaosylceramide in plasma and urine by mass spectrometry. Clin Chem Lab Med 2010;48:189–98.10.1515/CCLM.2010.048Suche in Google Scholar PubMed

7. Krüger R, Tholey A, Jakoby T, Vogelsberger R, Mönnikes R, Rossmann H, et al. Quantification of the Fabry marker lysoGb3 in human plasma by tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2012;883–884:128–35.10.1016/j.jchromb.2011.11.020Suche in Google Scholar PubMed

8. Bauer SH, Wiechers MF, Bruns K, Przybylski M, Stuermer CA. Isolation and identification of the plasma membrane-associated intracellular protein reggie-2 from goldfish brain by chromatography and Fourier-transform ion cyclotron resonance mass spectrometry. Anal Biochem 2001;298:25–31.10.1006/abio.2001.5330Suche in Google Scholar PubMed

9. Jiwan JH, Wallemacq P, Hérent M. HPLC-high resolution mass spectrometry in clinical laboratory? Clin Biochem 2011;44:136–47.10.1016/j.clinbiochem.2010.08.018Suche in Google Scholar PubMed

10. Ojanperä I, Kolmonen M, Pelander A. Current use of high-resolution mass spectrometry in drug screening relevant to clinical and forensic toxicology and doping control. Anal Bioanal Chem 2012;403:1203–20.10.1007/s00216-012-5726-zSuche in Google Scholar PubMed

11. Ojanperä S, Pelander A, Pelzing M, Krebs I, Vuori E, Ojanperä I. Isotopic pattern and accurate mass determination in urine drug screening by liquid chromatography/time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 2006;20:1161–7.10.1002/rcm.2429Suche in Google Scholar PubMed

12. Guddat S, Solymos E, Orlovius A, Thomas A, Sigmund G, Geyer H, et al. High-throughput screening for various classes of doping agents using a new ‘dilute-and-shoot’ liquid chromatography-tandem mass spectrometry multi-target approach. Drug Test Analysis 2011;3:836–50.10.1002/dta.372Suche in Google Scholar PubMed

13. Polettini A, Gottardo R, Pascali JP, Tagliaro F. Implementation and performance evaluation of a database of chemical formulas for the screening of pharmaco/toxicologically relevant compounds in biological samples using electrospray ionization-time-of-flight mass spectrometry. Anal Chem 2008;80:3050–7.10.1021/ac800071nSuche in Google Scholar PubMed

14. Zhu M, Zhang H, Humphreys WG. Drug metabolite profiling and identification by high-resolution mass spectrometry. J Biol Chem 2011;286:25419–25.10.1074/jbc.R110.200055Suche in Google Scholar PubMed PubMed Central

15. Hopfgartner G, Tonoli D, Varesio E. High-resolution mass spectrometry for integrated qualitative and quantitative analysis of pharmaceuticals in biological matrices. Anal Bioanal Chem 2012;402:2587–96.10.1007/s00216-011-5641-8Suche in Google Scholar PubMed

16. Ramanathan R, Jemal M, Ramagiri S, Xia Y, Humpreys WG, Olah T, et al. It is time for a paradigm shift in drug discovery bioanalysis: from SRM to HRMS. J Mass Spectrom 2011;46: 595–601.10.1002/jms.1921Suche in Google Scholar PubMed

17. Bruce SJ, Rochat B, Béguin A, Pesse B, Guessous I, Boulat O, et al. Analysis and quantification of vitamin D metabolites in serum by ultra-performance liquid chromatography coupled to tandem mass spectrometry and high-resolution mass spectrometry – a method comparison and validation. Rapid Commun Mass Spectrom 2013;27:200–6.10.1002/rcm.6439Suche in Google Scholar PubMed

18. Meyer MR, Helfer AG, Maurer HH. Current position of high-resolution MS for drug quantification in clinical & forensic toxicology. Bioanalysis 2014;6:2275–84.10.4155/bio.14.164Suche in Google Scholar PubMed

19. Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B. Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer for quantitative analyses of drugs. Rapid Commun Mass Spectrom 2012;26:499–509.10.1002/rcm.6121Suche in Google Scholar PubMed

20. Kaufmann A, Widmer M, Maden K. Post-interface signal suppression. A phenomenon observed in a single-stage orbitrap mass spectrometer coupled to an electrospray interfaced liquid chromatograph. Rapid Commun Mass Spectrom 2010;24:2162–70.10.1002/rcm.4615Suche in Google Scholar PubMed

21. Koster RA, Dijkers EC, Uges DR. Robust, high-throughput LC-MS/MS method for therapeutic drug monitoring of cyclosporine, tacrolimus, everolimus, and sirolimus in whole blood. Ther Drug Monit 2009;31:116–25.10.1097/FTD.0b013e318192304cSuche in Google Scholar PubMed

22. CLSI. Evaluation of the linearity of quantitative measurement procedures: A statistical approach; approved guideline. CLSI document EP06-A. Wayne, PA: NCCLS, 2003.Suche in Google Scholar

23. CLSI. Evaluation of precision performance of quantitative measurement methods; approved guideline. CLSI document EP05-A2. Wayne, PA: NCCLS, 2004.Suche in Google Scholar

24. CLSI. Method comparison and bias estimation using patient samples; approved guideline. CLSI-document EP09-A2. Wayne, PA: NCCLS, 2002.Suche in Google Scholar

Received: 2015-9-4
Accepted: 2015-10-28
Published Online: 2015-12-7
Published in Print: 2016-7-1

©2016 by De Gruyter

Artikel in diesem Heft

  1. Frontmatter
  2. In Memoriam: Gérard Siest (1936–2016)
  3. Editorials
  4. Improving diagnosis and reducing diagnostic errors: the next frontier of laboratory medicine
  5. The quality indicator paradox
  6. Review
  7. Clinical utility of the (-2)proPSA and evaluation of the evidence: a systematic review
  8. Mini Review
  9. Why are clinical practice guidelines not followed?
  10. EFLM Position Paper
  11. Patient identification and tube labelling – a call for harmonisation
  12. Genetics and Molecular Diagnostics
  13. A fast and simple method for detecting and quantifying donor-derived cell-free DNA in sera of solid organ transplant recipients as a biomarker for graft function
  14. Performance of two commercially available BCR-ABL1 quantification assays that use an international reporting scale
  15. HAND1 loss-of-function mutation associated with familial dilated cardiomyopathy
  16. General Clinical Chemistry and Laboratory Medicine
  17. Performance criteria and quality indicators for the post-analytical phase
  18. Assessing the commutability of reference material formats for the harmonization of amyloid-β measurements
  19. Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
  20. Measurement of plasma vitamin K1 (phylloquinone) and K2 (menaquinones-4 and -7) using HPLC-tandem mass spectrometry
  21. Therapeutic drug monitoring of infliximab: performance evaluation of three commercial ELISA kits
  22. High level of oxysterols in neonatal cholestasis: a pitfall in analysis of biochemical markers for Niemann-Pick type C disease
  23. Reference Values and Biological Variations
  24. Reference intervals of plasma homoarginine from the German Gutenberg Health Study
  25. TSH and fT4 during pregnancy: an observational study and a review of the literature
  26. Cancer Diagnostics
  27. Mean corpuscular volume levels and all-cause and liver cancer mortality
  28. Cardiovascular Diseases
  29. Circulating endothelial-derived apoptotic microparticles and insulin resistance in non-diabetic patients with chronic heart failure
  30. Hematology and Coagulation
  31. Platelet aggregation in response to ADP is highly variable in normal donors and patients on anti-platelet medication
  32. Letters to the Editor
  33. EQA-derived metrics to assess overall instrument performance
  34. Misidentification in laboratory medicine and diagnostic process: a neglected problem calling for action
  35. Impact of the routine implementation of automated indirect immunofluorescence antinuclear antibody analysis: 1 year of experience
  36. The importance of angiogenic markers in the differential diagnosis of HELLP syndrome vs. non-HELLP thrombocytopenia
  37. Bisalbuminemia accompanying bisalbuminuria detected in capillary electrophoresis, not in gel electrophoresis
  38. Prognostic value of red blood cell distribution width in acute pancreatitis patients admitted to intensive care units: an analysis of a publicly accessible clinical database MIMIC II
  39. Differences in analytical and biological results between older and newer lots of a widely used irisin immunoassay question the validity of previous studies
  40. A diagnostic algorithm for the detection of inhibitors against coagulation Factor V
  41. Interference of anticoagulants on coagulation testing
  42. Congress Abstracts
  43. Swiss MedLab 2016 and 74th Annual Meeting of the Swiss Society of Microbiology SSM, Bern, 13–16 June 2016
https://doi.org/10.1515/cclm-2015-0863
Schlagwörter für diesen Artikel
high resolution mass spectrometry; immunosuppressants; LC-MS/MS; method validation; TOF-MS

Artikel in diesem Heft

  1. Frontmatter
  2. In Memoriam: Gérard Siest (1936–2016)
  3. Editorials
  4. Improving diagnosis and reducing diagnostic errors: the next frontier of laboratory medicine
  5. The quality indicator paradox
  6. Review
  7. Clinical utility of the (-2)proPSA and evaluation of the evidence: a systematic review
  8. Mini Review
  9. Why are clinical practice guidelines not followed?
  10. EFLM Position Paper
  11. Patient identification and tube labelling – a call for harmonisation
  12. Genetics and Molecular Diagnostics
  13. A fast and simple method for detecting and quantifying donor-derived cell-free DNA in sera of solid organ transplant recipients as a biomarker for graft function
  14. Performance of two commercially available BCR-ABL1 quantification assays that use an international reporting scale
  15. HAND1 loss-of-function mutation associated with familial dilated cardiomyopathy
  16. General Clinical Chemistry and Laboratory Medicine
  17. Performance criteria and quality indicators for the post-analytical phase
  18. Assessing the commutability of reference material formats for the harmonization of amyloid-β measurements
  19. Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
  20. Measurement of plasma vitamin K1 (phylloquinone) and K2 (menaquinones-4 and -7) using HPLC-tandem mass spectrometry
  21. Therapeutic drug monitoring of infliximab: performance evaluation of three commercial ELISA kits
  22. High level of oxysterols in neonatal cholestasis: a pitfall in analysis of biochemical markers for Niemann-Pick type C disease
  23. Reference Values and Biological Variations
  24. Reference intervals of plasma homoarginine from the German Gutenberg Health Study
  25. TSH and fT4 during pregnancy: an observational study and a review of the literature
  26. Cancer Diagnostics
  27. Mean corpuscular volume levels and all-cause and liver cancer mortality
  28. Cardiovascular Diseases
  29. Circulating endothelial-derived apoptotic microparticles and insulin resistance in non-diabetic patients with chronic heart failure
  30. Hematology and Coagulation
  31. Platelet aggregation in response to ADP is highly variable in normal donors and patients on anti-platelet medication
  32. Letters to the Editor
  33. EQA-derived metrics to assess overall instrument performance
  34. Misidentification in laboratory medicine and diagnostic process: a neglected problem calling for action
  35. Impact of the routine implementation of automated indirect immunofluorescence antinuclear antibody analysis: 1 year of experience
  36. The importance of angiogenic markers in the differential diagnosis of HELLP syndrome vs. non-HELLP thrombocytopenia
  37. Bisalbuminemia accompanying bisalbuminuria detected in capillary electrophoresis, not in gel electrophoresis
  38. Prognostic value of red blood cell distribution width in acute pancreatitis patients admitted to intensive care units: an analysis of a publicly accessible clinical database MIMIC II
  39. Differences in analytical and biological results between older and newer lots of a widely used irisin immunoassay question the validity of previous studies
  40. A diagnostic algorithm for the detection of inhibitors against coagulation Factor V
  41. Interference of anticoagulants on coagulation testing
  42. Congress Abstracts
  43. Swiss MedLab 2016 and 74th Annual Meeting of the Swiss Society of Microbiology SSM, Bern, 13–16 June 2016
Jetzt anmelden und 20% sparen
Institutioneller Zugang
Wie funktioniert Authentifizierung?
Haben Sie eine Idee, wie wir unsere Website verbessern können?
Bitte schreiben Sie uns.
© 2025 De Gruyter Brill
Heruntergeladen am 22.9.2025 von https://www.degruyterbrill.com/document/doi/10.1515/cclm-2015-0863/html
Button zum nach oben scrollen