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FG‐4592 is a hypoxia‐inducible factor inhibitor that has been approved for therapeutic use in some countries. This class of compounds can increase the oxygen carrying capacity of the blood and thus have the potential to be used as performance enhancing agents in sports. The purpose of this study was to investigate the detection of FG‐4592 and metabolites in equine plasma and mane hair following a multiple dose oral administration to two Thoroughbred racehorses, to identify the best analytical targets for doping control laboratories. Urine samples were also analysed, and the results compared to previously published urine data. Liquid chromatography‐high resolution mass spectrometry was used for metabolite identification in urine and plasma. Liquid chromatography–tandem mass spectrometry was used for full sample analysis of urine, plasma and hair samples and generation of urine and plasma profiles. FG‐4592 and a mono‐hydroxylated metabolite were detected in plasma. FG‐4592 was detected with the greatest abundance and gave the longest duration of detection, up to 312 h post‐administration, and would be the recommended target in routine doping samples. FG‐4592 was detected in all mane hair samples collected post‐administration, up to 166 days following the final dose, showing extended detection can be achieved with this matrix. To the best of the authors' knowledge, this is the first report of FG‐4592 and metabolites in equine plasma and hair samples. Urine results were consistent with the previously published data, with FG‐4592 offering the best target for detection and longest detection periods. [ABSTRACT FROM AUTHOR]

Gene doping in horses is a threat to the fairness in sport and has serious implications for animal welfare. To investigate the effect of long‐term storage on the detection of AAV in plasma and whole blood, samples from an administration study using an adeno‐associated virus serotype 6 expressing green fluorescence protein (AAV6‐GFP) were stored at −20°C for 8 months before analysis. The AAV vector was detected in stored plasma samples, following the same detection profile as the fresh plasma samples. The stored blood showed lower overall DNA detection but followed the same detection profile as the plasma samples. This study provides confidence that re‐analysing plasma samples and/or analysing a frozen 'B' sample with different matrix such as whole blood after prolonged storage will still result in the detection of gene doping material. [ABSTRACT FROM AUTHOR]

Boldenone is an anabolic–androgenic steroid (AAS) that is prohibited in equine sports. However, in certain situations, it is endogenous, potentially formed by the microbes in urine. An approach to the differentiation based on the detection of the biomarkers Δ1‐progesterone, 20(S)‐hydroxy‐Δ1‐progesterone and 20(S)‐hydroxyprogesterone was assessed, and their concentrations were monitored in the urine of untreated female horses (n = 291) alongside boldenone, boldienone, testosterone and androstenedione. Using an ultra‐sensitive analytical method, boldenone (256 ± 236 pg/mL, n = 290) and the biomarkers (Δ1‐progesterone up to 57.6 pg/mL, n = 8; 20(S)‐hydroxy‐Δ1‐progesterone 85.3 ± 181 pg/mL, n = 130; 20(S)‐hydroxyprogesterone 43.5 ± 92.1 pg/mL, n = 158) were detected at low concentrations. The ex vivo production of Δ1‐steroids was artificially induced following the storage of urine samples at room temperature for 7 days in order to assess the concentrations and ratios of the monitored steroids. The administration of inappropriately stored feed source also resulted in an increase in 20(S)‐hydroxy‐Δ1‐progesterone concentrations and the biomarker ratios. Using the results from different datasets, an approach to differentiation was developed. In situations where the presence of boldenone exceeds a proposed action limit of 5 ng/mL, the presence of the biomarkers would be investigated. If Δ1‐progesterone is above 50 pg/mL or if 20(S)‐hydroxy‐Δ1‐progesterone is above 100 pg/mL with the ratio of 20(S)‐hydroxy‐Δ1‐progesterone:20(S)‐hydroxyprogesterone greater than 5:1, then this would indicate ex vivo transformation or consumption of altered feed rather than steroid administration. There remains a (small) possibility of a false negative result, but the model increases confidence that adverse analytical findings reported in female horses are caused by AAS administrations. [ABSTRACT FROM AUTHOR]

One male and one female giant panda (Ailuropoda melanoleuca) from a Belgian zoo were anesthetized on eight different occasions over a course of 4 yr for electro-ejaculation (n = 3) or artificial insemination (n = 5). Medetomidine (0.03–0.04 mg/kg) and ketamine (2.5–3 mg/kg) were administered by intramuscular remote injection. Animals gained sternal recumbency with the loss of response to external stimuli after 4.9 ± 1.6 min (mean ± SD). The trachea was intubated with a 14-mm–internal diameter endotracheal tube; anesthesia was maintained with isoflurane in oxygen adjusted according to the required depth of anesthesia with a small-animal circle system. Physiological variables (heart rate, respiratory rate, oxygenation, end tidal carbon dioxide partial pressure and non-invasive blood pressure) were measured and remained within an acceptable range throughout anesthesia. Atipamezole (0.17–0.25 mg/kg) was administered intramuscularly after anesthesia. Recoveries were rapid and uneventful. Medetomidine 0.03 mg/kg and ketamine 2.5 mg/kg IM appeared to be the preferred doses for giant pandas. [ABSTRACT FROM AUTHOR]

MK‐0677 (ibutamoren) is an orally active non‐peptide growth hormone secretagogue that binds to the ghrelin receptor stimulating the secretion of endogenous growth hormone. It is one of the most prevalent performance‐enhancing compounds currently available online and is potentially subject to abuse both in human and equine sports. The aim of the current study was to investigate whether it could be detected in equine hair following oral administration of MK‐0677 mesylate to a Thoroughbred racehorse. MK‐0677 and its O‐dealkylated metabolite were extracted using an existing method for prohibited substances in equine hair and analysed by liquid chromatography tandem mass spectrometry. This enabled the detection of MK‐0677 in all hair samples collected, up to 209 days in mane and 358 days in tail. A follow‐up methodology with an extensive wash procedure was carried out for selected hair samples, which unambiguously verified the presence of MK‐0677. Wash criteria to differentiate between internal incorporation (via bloodstream) and external deposition (via sweat and sebum) was also assessed and indicated internal incorporation for the samples collected at later time points (≥52 days) and a combination of internal incorporation and external deposition for hair samples collected at the earlier time point (2 days). [ABSTRACT FROM AUTHOR]