Validation of a Quantitative Multi-Residue Urinary Assay for the Detection of Androgen, Oestrogen and Progestagen Abuse in the Bovine

Scarth, James; Clarke, Adam; Hands, Jonathan; Teale, P.; Macarthur, Roy; Kay, Jack F.

doi:10.1365/s10337-009-1429-9

Cited by 21 publications

(26 citation statements)

References 9 publications

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“…This meant that it was not possible to design a recovery experiment combining free and conjugated steroid at a ratio relevant to all situations. Therefore, the b-glucuronidase enzyme was added in excess (as demonstrated in [23]) in order to ensure that maximal quantities of steroid conjugate were hydrolysed. Furthermore, the proportion of free versus glucuronide conjugated steroid in each sample was not determined because there was typically insufficient sample volume to carry out multiple analyses (with and without hydrolysis).…”

Section: Recoverymentioning

confidence: 99%

Detection of Nandrolone and Boldenone Abuse in the Ovine by GC–MS–MS

Clarke¹,

Scarth²,

Hands³

et al. 2011

Chromatographia

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Under European legislation, the use of anabolic steroids as growth promoters in meat production is prohibited. Currently, there is no internationally accepted method used for the detection of the potentially endogenous steroids nandrolone and boldenone in the ovine. In the current study, a multi-residue GC-MS-MS-based urinary assay has been validated for boldenone as well as the nandrolone metabolites 5a-estrane-3b,17a-diol and epinandrolone. Using a standard addition calibration line approach in pooled bovine urine, the method was linear between the endogenous concentrations and those augmented with 6,000 pg mL -1 . The method was then applied to populations of wether (n = 242) and ewe (n = 237) ovine animals in order to establish urinary thresholds for detecting nandrolone and boldenone abuse. A statistical model (the Chebyshev inequality) was used to produce threshold concentrations for each analyte. Adjustment of the nandrolone metabolite data for specific gravity, a measure of the hydration status of the animal, allowed the effective thresholds to be reduced; potentially leading to a lower number of false positives. Furthermore, the proposed epinandrolone confirmatory thresholds (38,628 and 57,950 pg mL -1 in wethers and ewes, respectively) were found to be effective in detecting abuse of nandrolone for at least 1 month post-dose of this steroid. However, further studies would be required to assess the efficacy of the proposed boldenone confirmatory thresholds (19,857 and 56,080 pg mL -1 in wethers and ewes, respectively) since data on its excretion following administration to the ovine are lacking.

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Section: Recoverymentioning

confidence: 99%

Detection of Nandrolone and Boldenone Abuse in the Ovine by GC–MS–MS

Clarke¹,

Scarth²,

Hands³

et al. 2011

Chromatographia

Self Cite

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“…-Can be combined with some other screening approaches such as threshold concentrations if the same analytical technique is used. Threshold concentrations (urine, plasma/serum) [42][43][44][46][47][48] -Can be used for both screening and confirmation by using different thresholds.…”

Section: Discussion and Future Perspectivesmentioning

confidence: 99%

“…The first use of the 'y out of n' approach involved the proposal of urinary concentration thresholds for detecting the abuse of nandrolone, boldenone, testosterone and oestradiol abuse in cattle [46,47] For each analyte, the use of the '2 out of 2' and '3 out of 3' animal approaches lead to concentration thresholds approximately 10-and 20-fold lower, respectively, compared with the traditional '1 out of 1' (single animal) approach. However, although the larger 'y out of n' approach allows for a reduction in the concentration threshold, the risk of false compliances increases with n due to potential inter-individual variation in excretion profiles.…”

Section: Threshold Concentrationsmentioning

confidence: 99%

A review of analytical strategies for the detection of ‘endogenous’ steroid abuse in food production

Scarth¹,

Kay

Teale³

et al. 2012

Drug Testing and Analysis

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Detection of the abuse of synthetic steroids in food production is nowadays relatively straightforward using modern techniques such as gas or liquid chromatography coupled to mass spectrometry (GC-MS/MS or LC-MS/MS, respectively). However, proving the abuse of 'endogenous' (or naturally occurring) steroids is more difficult. Despite these difficulties, significant progress in this area has recently been made and a number of methods are now available.The aim of the current review was to systematically review the available analytical approaches, which include threshold concentrations, qualitative 'marker' metabolites, intact steroid esters, gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS), longitudinal testing and 'omics' biomarker profiling. The advantages/disadvantages of these methods are considered in detail, but the choice of which to adopt is dictated by a number of practical, political, and economic factors, which vary in different parts of the world. These include the steroid/species combination requiring analysis, the matrix tested, whether samples are collected from live or slaughtered animals, available analytical instrumentation, sample throughput/cost, and the relevant legal/regulatory frameworks. Furthermore, these approaches could be combined in a range of different parallel and/or sequential screening/confirmatory testing streams, with the final choice being determined by the aforementioned considerations.Despite these advances, more work is required to refine the different techniques and to respond to the ever increasing list of compounds classified as 'endogenous'. At this advanced stage, however, it is now more important than ever for scientists and regulators from across the world to communicate and collaborate in order to harmonize and streamline research efforts.

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“…The generated derivatives were sensitively analyzed by LC/ESI-MS/MS (11). DnsCl is widely used for the analysis of the compounds having hydroxyl group (10)(11)(12)(13)(14)(15)(16)(17)(18)(19). Similarly, picolinic acid (20-26), fusaric acid (27), dimethylglycine (28), isonicotinoyl azide (29,30), and N-methyl-nicotinic acid N-hydroxysuccinimide ester (C1-NA-NHS) (31) were used for the derivatization of alcohols and phenols.…”

Section: Alcohols and Phenolsmentioning

confidence: 99%

Derivatization in liquid chromatography for mass spectrometric detection

Santa¹

2013

Drug Discov Ther

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Validation of a Quantitative Multi-Residue Urinary Assay for the Detection of Androgen, Oestrogen and Progestagen Abuse in the Bovine

Cited by 21 publications

References 9 publications

Detection of Nandrolone and Boldenone Abuse in the Ovine by GC–MS–MS

Detection of Nandrolone and Boldenone Abuse in the Ovine by GC–MS–MS

A review of analytical strategies for the detection of ‘endogenous’ steroid abuse in food production

Derivatization in liquid chromatography for mass spectrometric detection

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