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LC-MS/MS fast analysis of androgenic steroids in urine

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Abstract

The liquid chromatography–tandem mass spectrometry method (LC-MS/MS) was developed and validated to detect androgenic steroids: trenbolone, nortestosterone, boldenone, methylboldenone, testosterone, methyltestosterone, 17β-1-testosterone and their metabolites in bovine urine. Sample preparation before LC-MS/MS analysis involved an enzymatic hydrolysis with glucuronidase AS-HP, isolation of free hormones from urine on C18 SPE column and purification of the extract using liquid–liquid extraction with n-pentane and SPE NH2 column. For the chromatographic separation of steroids, the Poroshell 120-EC C18 column (150 × 2.1 mm, 2.7 μm) was used. Mass spectrometric measurement was achieved using the API 4000 triple quadrupole (QqQ) instrument with a TurboIon-Spray source operating in positive electrospray ionization mode. The procedure was validated according to the Decision 2002/657/EC. Recovery ranged from 76.5% to 118.9% for all examined compounds. The repeatability was below 20% and reproducibility did not exceed the 25%. The linearity was good for all analytes in the whole range of tested concentrations, as proved by the correlation coefficients greater than 0.99. The decision limit (CCα) ranged from 0.10 to 0.17 μg L−1 for all analytes, whereas the detection capability (CCβ) ranged from 0.17 to 0.29 μg L−1. The application of an innovative Poroshell column allowed for very good chromatographic separation of steroids with a much shorter time of analysis.

MRM chromatogram of 30 ng of androgenic steroids

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100, Pulawy, Poland

    Barbara Wozniak, Iwona Matraszek-Zuchowska & Jan Zmudzki

Authors
  1. Barbara Wozniak
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  2. Iwona Matraszek-Zuchowska
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  3. Jan Zmudzki
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Corresponding author

Correspondence to Barbara Wozniak.

Additional information

Published in the special paper collection Recent Advances in Food Analysis with guest editors J. Hajslova, R. Krska, M. Nielen.

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Wozniak, B., Matraszek-Zuchowska, I. & Zmudzki, J. LC-MS/MS fast analysis of androgenic steroids in urine. Anal Bioanal Chem 403, 2965–2972 (2012). https://doi.org/10.1007/s00216-012-5859-0

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  • DOI: https://doi.org/10.1007/s00216-012-5859-0

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