Advanced search
Add to list

Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry

Author
Organization
Abstract
For over a decade there has been an intensive debate on the possible natural origin of boldenone (androst-1,4-diene-17 beta-ol-3-one, 17 beta-boldenone) in calf urine and several alternative markers to discriminate between endogenously formed boldenone and exogenously administered boldenone have been suggested. The currently approved method for proving illegal administration of beta-boldenone(ester) is the detection of P-boldenone conjugates. In the presented method the sulphate, glucuronide and free fractions are separated from each other during cleanup on a SAX column to be able to determine the conjugated status of the boldenone metabolites. The sulphate and glucuronide fractions are submitted to hydrolysis and all three fractions are further cleaned up on a combination of C-18/NH2 solid-phase extraction (SPE) columns. Chromatographic separation of the boldenone metabolites was achieved with a Waters Acquity UPLC (TM) instrument using a Sapphire C-18 (1.7 mu m; 2 x 50 mm) column within 5 min. Detection of the analytes was achieved by electrospray ionisation tandem mass spectrometry. The decision limits of this method, validated according to Commission Decision 2002/657/EC, were 0.08 ng mL(-1) for androsta-1,4-diene-3,17-dione, 0.13 ng mL(-1) for androst4-ene-3,17-dione, 0.11 ng mL(-1) for 17 alpha-boldenone, 0.07 ng mL(-1) for 17 beta-boldenone, 0.24 ng mL(-1) for 5 beta-androst-1-en-17 beta-ol-3-one and 0.58 ng mL(-1) for 6 beta-hydroxy-17 beta-boldenone. Because of the fractionation approach used in this method there is no need for conjugated reference standards which often are not available. The disadvantage of needing three analytical runs to determine the conjugated status of each of the metabolites was overcome by using fast chromatography.
Keywords
ANIMAL FEED, BOVINE URINE, VEAL CALVES, CATTLE, FECES, ANDROSTA-1, 4-DIENE-3, GROWTH PROMOTERS, 17-ALPHA-BOLDENONE, 17-DIONE, 17-BETA-BOLDENONE, IDENTIFICATION

Citation

Please use this url to cite or link to this publication:

MLA
Van Poucke, Christof, Evy Van Vossel, and Carlos Van Peteghem. “Fractionation of Free and Conjugated Steroids for the Detection of Boldenone Metabolites in Calf Urine with Ultra-performance Liquid Chromatography/tandem Mass Spectrometry.” RAPID COMMUNICATIONS IN MASS SPECTROMETRY 22.15 (2008): 2324–2332. Print.
APA
Van Poucke, C., Van Vossel, E., & Van Peteghem, C. (2008). Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 22(15), 2324–2332.
Chicago author-date
Van Poucke, Christof, Evy Van Vossel, and Carlos Van Peteghem. 2008. “Fractionation of Free and Conjugated Steroids for the Detection of Boldenone Metabolites in Calf Urine with Ultra-performance Liquid Chromatography/tandem Mass Spectrometry.” Rapid Communications in Mass Spectrometry 22 (15): 2324–2332.
Chicago author-date (all authors)
Van Poucke, Christof, Evy Van Vossel, and Carlos Van Peteghem. 2008. “Fractionation of Free and Conjugated Steroids for the Detection of Boldenone Metabolites in Calf Urine with Ultra-performance Liquid Chromatography/tandem Mass Spectrometry.” Rapid Communications in Mass Spectrometry 22 (15): 2324–2332.
Vancouver
1.
Van Poucke C, Van Vossel E, Van Peteghem C. Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY. 2008;22(15):2324–32.
IEEE
[1]
C. Van Poucke, E. Van Vossel, and C. Van Peteghem, “Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry,” RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 22, no. 15, pp. 2324–2332, 2008.
@article{533785,
  abstract     = {{For over a decade there has been an intensive debate on the possible natural origin of boldenone (androst-1,4-diene-17 beta-ol-3-one, 17 beta-boldenone) in calf urine and several alternative markers to discriminate between endogenously formed boldenone and exogenously administered boldenone have been suggested. The currently approved method for proving illegal administration of beta-boldenone(ester) is the detection of P-boldenone conjugates. In the presented method the sulphate, glucuronide and free fractions are separated from each other during cleanup on a SAX column to be able to determine the conjugated status of the boldenone metabolites. The sulphate and glucuronide fractions are submitted to hydrolysis and all three fractions are further cleaned up on a combination of C-18/NH2 solid-phase extraction (SPE) columns. Chromatographic separation of the boldenone metabolites was achieved with a Waters Acquity UPLC (TM) instrument using a Sapphire C-18 (1.7 mu m; 2 x 50 mm) column within 5 min. Detection of the analytes was achieved by electrospray ionisation tandem mass spectrometry. The decision limits of this method, validated according to Commission Decision 2002/657/EC, were 0.08 ng mL(-1) for androsta-1,4-diene-3,17-dione, 0.13 ng mL(-1) for androst4-ene-3,17-dione, 0.11 ng mL(-1) for 17 alpha-boldenone, 0.07 ng mL(-1) for 17 beta-boldenone, 0.24 ng mL(-1) for 5 beta-androst-1-en-17 beta-ol-3-one and 0.58 ng mL(-1) for 6 beta-hydroxy-17 beta-boldenone. Because of the fractionation approach used in this method there is no need for conjugated reference standards which often are not available. The disadvantage of needing three analytical runs to determine the conjugated status of each of the metabolites was overcome by using fast chromatography.}},
  author       = {{Van Poucke, Christof and Van Vossel, Evy and Van Peteghem, Carlos}},
  issn         = {{0951-4198}},
  journal      = {{RAPID COMMUNICATIONS IN MASS SPECTROMETRY}},
  keywords     = {{ANIMAL FEED,BOVINE URINE,VEAL CALVES,CATTLE,FECES,ANDROSTA-1,4-DIENE-3,GROWTH PROMOTERS,17-ALPHA-BOLDENONE,17-DIONE,17-BETA-BOLDENONE,IDENTIFICATION}},
  language     = {{eng}},
  number       = {{15}},
  pages        = {{2324--2332}},
  title        = {{Fractionation of free and conjugated steroids for the detection of boldenone metabolites in calf urine with ultra-performance liquid chromatography/tandem mass spectrometry}},
  url          = {{http://dx.doi.org/10.1002/rcm.3617}},
  volume       = {{22}},
  year         = {{2008}},
}

Altmetric
View in Altmetric
Web of Science
Times cited: