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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
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1437-4331
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Volume 49, Issue 7

Issues

High-throughput LC-MS/MS method for monitoring sirolimus and everolimus in the routine clinical laboratory

Mariela Ivanova / Carlo Artusi / Giulia Polo / Martina Zaninotto / Mario Plebani
  • Department of Laboratory Medicine, University-Hospital, Padova, Italy
  • Leonardo Foundation, Abano Terme General Hospital, Padova, Italy
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2011-05-17 | DOI: https://doi.org/10.1515/CCLM.2011.192

Abstract

Background: Immunosuppressant therapeutic drug monitoring (TDM) is an important requirement in post-transplant patient care. In recent years, high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has become a valid alternative to antibody-based immunoassays in TDM due to its high specificity and sensitivity. Furthermore, this technology allows for the simultaneous measurement of several immunosuppressive drugs. The aim of the present study was to establish a straightforward, robust, and high-throughput LC-MS/MS method for the simultaneous determination of sirolimus and everolimus in whole blood in order to replace immunoassays in our routine practice.

Methods: Five-level blood calibrators were employed for assay development, while three materials at different concentrations were used for internal quality control. The proposed method uses protein precipitation for sample preparation. Analyses were performed using a triple quadrupole LC-MS/MS, with a C18 held at 60°C. Using an appropriate gradient elution profile and SPE on-line, elution times for all compounds analysed were 2.6 min with a total run-time of 3.5 min.

Results: Calibration curves were linear throughout the selected ranges. The intra- and inter-assay CVs (<7%), the limit of quantification (0.2 μg/L) and accuracy were highly satisfactory. On testing the results using the international proficiency testing scheme (UK-NEQAS), the performance of the proposed method was found to be highly reliable.

Conclusions: The findings made by us indicate that the proposed method is of value, since it is speedy, straightforward, accurate, and applicable to different LC-MS/MS instruments for the routine TDM of organ transplant recipients.

Keywords: everolimus; immunosuppressants; liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS); mass spectrometry; sirolimus

About the article

Corresponding author: Martina Zaninotto, Department of Laboratory Medicine, University-Hospital of Padova, Via Giustiniani 2, 35128 Padova, Italy Phone: +39-049-8213230, Fax: +39-049-8218489


Received: 2010-11-25

Accepted: 2011-02-18

Published Online: 2011-05-17

Published in Print: 2011-07-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 49, Issue 7, Pages 1151–1158, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2011.192.

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