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Journal of Veterinary Research

formerly Bulletin of the Veterinary Institute in Pulawy

4 Issues per year

IMPACT FACTOR Bull Vet Inst Pulawy 2016: 0.462

CiteScore 2016: 0.46

SCImago Journal Rank (SJR) 2015: 0.230
Source Normalized Impact per Paper (SNIP) 2015: 0.383

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Development of LC-MS/MS confirmatory method for the determination of testosterone in bovine serum

Barbara Woźniak / Iwona Matraszek-Żuchowska
  • Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland
  • Other articles by this author:
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/ Sebastian Witek
  • Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Andrzej Posyniak
  • Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2017-04-04 | DOI: https://doi.org/10.1515/jvetres-2017-0010


Introduction: In the European Union the use of steroid growth promoters is prohibited under Council Directive 96/22/EC. For effective control of illegal use of natural steroids, highly sensitive analytical methods are required, because sex hormones can be present in very low concentrations in biological samples. The aim of the study was to develop a confirmatory method for the detection of testosterone in bovine serum at ppt level.

Material and Methods: 17β-testosterone and internal standards of 17β-testosterone-d2 were extracted from serum samples with a mixture of tert-butyl methyl ether/petroleum ether and were directly analysed by an LC/MS/MS on QTRAP 5500 instrument with a TurboIon-Spray source operating in a positive ionisation mode. Chromatographic separation was achieved on the analytical column Inertsil® ODS-3 with an isocratic elution using mobile phase consisting of acetonitrile, methanol, and water. Method validation has been carried out in accordance with the Commission Decision 2002/657/EC.

Results: The method was characterised by good recovery (82%) and precision (R.S.D 17 %). Decision limit (CCα) and detection capability (CCβ) was 0.05 μg L−1 and 0.09 μg L−1 respectively. The method met the criteria set out in Commission Decision 2002/657/EC for the purpose of confirmation in terms of retention time and ion ratio in the whole range of its application.

Conclusions: The developed method is specific and sensitive, suitable for measuring the natural level of testosterone in blood of cattle and for use in routine control programme for the detection of this hormone in bovine serum.

Keywords: testosterone; bovine serum; validation; liquid chromatography; mass spectrometry


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About the article

Received: 2016-08-23

Accepted: 2017-03-10

Published Online: 2017-04-04

Published in Print: 2017-03-01

Citation Information: Journal of Veterinary Research, ISSN (Online) 2450-8608, DOI: https://doi.org/10.1515/jvetres-2017-0010.

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© 2017 Barbara Woźniak et al., published by De Gruyter Open. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

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