An in vitro study on metabolism of 17β-boldenone and boldione using cattle liver and kidney subcellular fractions

Merlanti, Roberta; Gallina, Guglielmo; Capolongo, Francesca; Contiero, Lidia; Biancotto, Giancarlo; Dacasto, Mauro; Montesissa, Clara

doi:10.1016/j.aca.2006.12.007

Cited by 9 publications

(12 citation statements)

References 14 publications

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“…Indeed, it is activated by the same widely distributed enzyme, namely 17b-hydroxysteroid dehydrogenase, which converts AED to testosterone. Recently, its role as both precursor and metabolite of boldenone has been confirmed in vitro using subcellular fractions obtained from calf liver [14,15]. Moreover, ADD is considered the direct precursor of 1,4androstadiene-17a-ol-3-one (a-Bol), the inactive epimer of b-Bol.…”

Section: Introductionmentioning

confidence: 98%

“…Both b-Bol and ADD are illegally used in racehorses and athletes to improve sport performance [15]. Because of the known properties displayed by natural and synthetic androgens to react with specific receptors and to promote protein synthesis at muscular level, both compounds during the last decades were widely used also in cattle breeding for meat production, due to their favourable effects on the animal growth and feed conversion.…”

Section: Introductionmentioning

confidence: 99%

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Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression

Draisci

Montesissa²,

Santamaria

et al. 2007

Proteomics

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Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-Q-MS/MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone alpha- and beta-epimers were detected in plasma and urine only within 2 and 24 h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE, MALDI-TOF-MS and muLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasma samples collected before treatment, was found upregulated even 36 h after hormone treatment. Extensive mass mapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed by Western blot analysis confirmed a significant and time dependent increase of this ApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine.

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Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression

Draisci

Montesissa²,

Santamaria

et al. 2007

Proteomics

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“…In vitro studies [23] of boldenone and boldione using cattle liver and kidney subcellular fractions described the formation of hydroxylated metabolites mainly in 6β and 16α positions and the reduction of boldione in 17 position, producing boldenone and 17α-boldenone. In addition to the previously described androst-1,4-dien-6β-ol-3,17-dione (6β-hydroxy-boldione) and androst-1,4-dien-6β,17β-diol-3-one (6β-hydroxy-boldenone) in human urine, [4] other hydroxylated metabolites were detected.…”

Section: Resultsmentioning

confidence: 99%

Metabolism of boldione in humans by mass spectrometric techniques: detection of pseudoendogenous metabolites

Torre

Curcio

Colamonici

et al. 2013

Drug Testing and Analysis

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Boldione is an anabolic androgenic steroid (AAS) related to boldenone, androstenedione, and testosterone bearing two double bonds in C1 and C4 positions. Boldione is rapidly transformed to the well-known AAS boldenone, being both compounds included in the list of prohibited substances and methods published yearly by the World Anti-Doping Agency (WADA). After the administration of boldione to a male volunteer, the already described urinary metabolites of boldenone produced after reduction in C4, oxydoreduction in C3 and C17, and hydroxylation have been detected. In addition, minor new metabolites have been detected and their structure postulated after mass spectrometric analyses. Finally, the reduction of the double bound in C1 produces metabolites identical to the endogenously produced ones. A method based on gas chromatography coupled to isotope ratio mass spectrometry (GC/C/IRMS) after a urine sample purification by high performance liquid chromatography (HPLC) permitted to confirm the main synthetic like boldione/boldenone metabolite (17β-hydroxy-5β-androst-1-en-3-one) and boldenone at trace levels (< 5 ng/mL) and then to establish its synthetic or endogenous origin, and to determine the exogenous origin of metabolites with the same chemical structure of the endogenous ones. The detection of pseudoendogenous androgens of synthetic origin partially overlapped boldenone and its main metabolite detection, being an additional proof of synthetic steroids misuse. By the use of IRMS, the correct evaluation of the modifications of the steroid profile after the administration of synthetic AAS that could be converted into endogenous like ones is possible.

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“…The in vitro metabolism of 17β-Bol and ADD confirmed that several hydroxylated derivatives could be produced in bovine liver microsomes, among those 6β-hydroxy-17β-Bol and 6β-hydroxy-ADD (4,22). Also, 17R-Bol incubated with bovine liver microsomes and the NADPH generating system led to ADD and to a hydroxylated metabolite identified as 6β-hydroxy-17R-Bol (23).…”

Section: Discussionmentioning

confidence: 99%

Boldenone, Boldione, and Milk Replacers in the Diet of Veal Calves: The Effects of Phytosterol Content on the Urinary Excretion of Boldenone Metabolites

Gallina

Ferretti

Merlanti

et al. 2007

J. Agric. Food Chem.

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Twenty-six veal calves were split into two groups and fed two milk replacers with a different content of phytosterols for 26 days; then, 14 calves (7 animals from each diet) were kept as controls and 12 calves (6 per diet) received daily, per os, a combination of 17beta-boldenone (17beta-Bol) and androsta-1,4-dien-3,17-dione (ADD) for 38 days. The urinary elimination of 17 alpha-/17beta-boldenone conjugates (17 alpha/beta-Bol) and androsta-1,4-dien-3,17-dione (ADD) was followed by liquid chromatography-tandem mass spectrometry from all of the animals until slaughtering. In urine from treated animals, 17 alpha-Bol concentrations, despite a great variability, were greater than 17beta-Bol, both detected always as conjugates. At days 1, 2, and 3, the mean urine concentration of 17 alpha-Bol was higher than 12 ng/mL. A remarkable decrease was observed during the following days, but the 17 alpha-Bol concentration was still higher than the attention level of 2 ng/mL in 58% of the samples; the concentration of 17beta-Bol was around the action level of 1 ng/mL; two days after treatment withdrawal, no 17beta-Bol was detected in the urine. In urine from control animals, the 17 alpha-Bol concentration was strictly related to the phytosterol content of the diet, while, in urine from treated animals, the much higher 17 alpha-Bol levels were not modified by the production from diet precursors. The results confirmed that a 17 alpha-Bol level higher than 2 ng/mL should be considered as evidence of suspected illegal treatment and that the urinary excretion of 17beta-Bol is due to exogenous administration of 17beta-Bol. The discontinuous rate of elimination of both 17 alpha- and 17beta-Bol, despite the daily administration of 17beta-Bol plus ADD, indicates the necessity for further research to detect other urinary boldenone metabolites to strength surveillance strategy.

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An in vitro study on metabolism of 17β-boldenone and boldione using cattle liver and kidney subcellular fractions

Cited by 9 publications

References 14 publications

Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression

Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression

Metabolism of boldione in humans by mass spectrometric techniques: detection of pseudoendogenous metabolites

Boldenone, Boldione, and Milk Replacers in the Diet of Veal Calves: The Effects of Phytosterol Content on the Urinary Excretion of Boldenone Metabolites

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