5α-Estrane-3β,17β-diol and 5β-estrane-3α,17β-diol: Definitive screening biomarkers to sign nandrolone abuse in cattle?

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.

Section: Discussion and Future Perspectivesmentioning

confidence: 99%

Section: Marker Metabolitesmentioning

confidence: 99%

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

Scarth¹,

Kay

Teale³

et al. 2012

“…In cattle, the metabolism of β-nandrolone involves the epimerization to α-nandrolone (Figure 1), which, as a sulphate or glucuronide conjugate, is the main urinary metabolite in this species. A number of other minor metabolites of β-nandrolone have been reported in cattle, namely 5α-estrane-3β, 17α-diol, 5β-oestrane-3α and 17β-diol, [12] or 19-noretiocholanolone and 19norepiandrosterone. A number of other minor metabolites of β-nandrolone have been reported in cattle, namely 5α-estrane-3β, 17α-diol, 5β-oestrane-3α and 17β-diol, [12] or 19-noretiocholanolone and 19norepiandrosterone.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of nandrolone and ractopamine in the urine of veal calves: liquid chromatography‐tandem mass spectrometry approach

Chiesa

Panseri

Cannizzo

et al. 2016

Research articles LC-MS/MS technology allows the direct detection of glucuronoconjugated metabolites. PI scan, NL scan and SRM methods, have demonstrated to be useful for the detection of unknown unpredicted and predicted glucuronide MTD metabolites in metabolic studies. Thirteen glucuronide metabolites were observed, and one of them was resistant to enzymatic hydrolysis, not detectable using the traditional GC-MS strategies. Sport supplements may contain steroids never approved for therapeutic use. In this study, 18 sport supplements purchased online underwent organic solvent extraction and subsequent isolated steroid tested for intrinsic androgen bioactivity. Many were found to be strong androgens. These potent pharmacologically active steroids are being used without regulatory control or consumer awareness of their potential adverse effect. Avidin-biotin technology was used for the implementation of an enzyme-linked immunosorbent assay (AB-ELISA) as a sensitive method for the detection of anabolic androgenic steroids (AAS) present in dietary supplements. Assembled AB-ELISAs were characterized and used for detection of various AAS in 49 samples of counterfeit dietary supplements. The concordance between UHPLC-MS and the CR corrected data from AB-ELISA indicated the potential of this method even to quantification of some steroids. Nandrolone (anabolic steroid) and ractopamine (β-adrenergic agonist) are used quite frequently in food-producing livestock, but little is known about their synergic action when they are administered together. The objective of this study was to determine levels after experimental treatment of bovines with both of the above-mentioned drugs. For this purpose two specific analytical methods based on liquid chromatography-tandem mass spectrometry were developed. The use of tandem yeast and mamma-lian cell in vitro androgen bioassays to detect androgens in internet-sourced sport supplements 545-552 We evaluated the effectiveness of drugs testing using hair and nails and the feasibility of determining when drugs were ingested by measuring the time courses of drug concentrations in hair and toenails after single administrations of various drugs. For most basic compounds, the maximum concentrations in hair segments tended to be higher than those in nails. Nails were an effective specimen to detect neutral and weakly acidic compounds. In this work, we aimed to develop a new, simple and confident sensory platform for cocaine detection. For the fabrication of this test system, surface of μ-well plates was modified with quantum dots (QDs) and then cocaine-binding aptamer modified gold nanoparticles (Apt-AuNP) were covalently attached to the QDs. Detectable signals were obtained owing to the target-induced conformational change of aptamer which gives rise to close interaction of QDs with their quencher, AuNPs. Hence, decrease in fluorescence intensity as a cocaine response was recorded. The developed test system-called as Aptamer Folding based Sensory Device, (AFSD)-was evaluated in terms of main a...

“…In this context, strategies based on the detection of physiological actions of anabolic practices are promising approaches to screen for their misuse. Profiling biological matrices to reveal biological effects of a drug can either be performed in a targeted focus on a particular class of compounds, such as the steroidome [4][5][6][7][8][9] or in an untargeted way using global strategies such as transcriptomics, proteomics, or metabolomics. [10,11] These emerging strategies are promising ways to highlight candidate biomarkers to tackle illegal practices.…”

Section: Introductionmentioning

confidence: 99%

Metabolomics in food analysis: application to the control of forbidden substances

Dervilly

Courant

Chéreau

et al. 2012

Self Cite

Metabolomics is a science of interest in food analysis to describe and predict properties of food products and processes. It includes the development of analytical methods with the ultimate goal being the identification of so-called 'quality markers', (i.e. sets of metabolites that correlate with, for example, quality, safety, taste, or fragrance of foodstuffs). In turn, these metabolites are influenced by factors as genetic differences of the raw food ingredients (such as animal breed or crop species differences), growth conditions (such as climate, irrigation strategy, or feeding) or production conditions (such as temperature, acidity, or pressure). In cases where the routine-based measurement of a food property faces some limitations such as the lack of knowledge regarding the target compounds to monitor, monitoring based on a limited set of crucial biomarkers is a good alternative, which is of great interest for food safety purposes regarding growth promoting practices. Such an approach may be more efficient than using a classic approach based on a limited set of known metabolites of anabolic compounds. In this context, screening strategies allowing detection of the physiological response resulting from anabolic compound administration are promising approaches to detect their misuse. The global metabolomics workflow implemented for such studies is presented and illustrated through various examples of biological matrices profiling (tissue, blood, urine) and for different classes of anabolic compounds (steroids, b-agonists and somatotropin).