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Licensed Unlicensed Requires Authentication Published by De Gruyter April 11, 2013

Quantification of vancomycin in human serum by LC-MS/MS

  • Katrin König EMAIL logo , Uwe Kobold , Gerhard Fink , Andreas Leinenbach , Thomas Dülffer , Roland Thiele , Johannes Zander and Michael Vogeser

Abstract

Background: The aim of our work was to develop and validate a reliable LC-MS/MS-based measurement procedure for the quantification of vancomycin in serum, to be applied in the context of efforts to standardize and harmonize therapeutic drug monitoring of this compound using routine assays.

Methods: Sample preparation was based on protein precipitation followed by ultrafiltration. In order to minimize differential modulation of ionization by matrix constituents extended chromatographic separation was applied leading to a retention time of 9.8 min for the analyte. Measurement was done by HPLC-ESI-MS/MS. For internal standardization the derivative vancomycin-glycin (ISTD) prepared by chemical synthesis was used, HPLC conditions ensured coelution of ISTD with the analyte.

Results: In a bi-center validation total CVs of <4% were observed for quality control material ranging from 5.3 mg/L to 79.4 mg/L; accuracy was ±4%. No relevant ion suppression was observed. Comparative measurement of aliquots from 70 samples at the two validation sites demonstrated close agreement.

Conclusions: Employing a closely related homologue molecule for internal standardization and the use of MS/MS following highly efficient sample pre-fractionation by HPLC, the method described here can be considered to offer the highest level of analytical reliability realized so far for the quantification of vancomycin in human serum. Thus, the method is suitable to be used in a comprehensive reference measurement system for vancomycin.


Corresponding author: Katrin König, Institute of Laboratory Medicine, Hospital of the University of Munich, Marchioninistrasse 15, 81375 Munich, Germany, Phone: +49 89 70953221

We thank the Referenzinstitut für Bioanalytik (RfB) for the permission to report the results of a vancomycin proficiency testing program.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

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Received: 2013-2-22
Accepted: 2013-3-21
Published Online: 2013-04-11
Published in Print: 2013-09-01

©2013 by Walter de Gruyter Berlin Boston

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