Presence of salicylic acid in Standardbred horse urine and plasma after various feed and drug administrations

Beaumier, P.; Fenwick, J. D.; Stevenson, A.; Weber, M. P.; Young, Larry M.

doi:10.1111/j.2042-3306.1987.tb01381.x

Cited by 8 publications

(7 citation statements)

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“…These threshold levels have been introduced to avoid positive cases due to salicylates originating from plants potentially present in equine feed, e.g. in alfalfa hay [7]. However, threshold levels might increase the risk of ASA being abused for therapeutic purposes up to the threshold in competition horses [8].…”

Section: Introductionmentioning

confidence: 99%

Synovial distribution of “systemically” administered acetylsalicylic acid in the isolated perfused equine distal limb

et al. 2013

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BackgroundThis study investigated synovial concentrations of acetylsalicylic acid (ASA) and its metabolite salicylic acid (SA) in the equine fetlock joint following systemic administration of ASA. Salicylates were chosen because SA is the only nonsteroidal anti-inflammatory drug for which threshold levels exist for plasma and urine in equine sports. To avoid animal experiments, the study was conducted using an ex vivo model of the isolated perfused equine distal limb in combination with plasma concentrations obtained from literature.Salicylate concentrations in the joint were determined using microdialysis and high performance liquid chromatography (HPLC). Any anti-inflammatory effect of synovial ASA concentrations was assessed using an ASA EC50 (half maximal effective concentration) determined in equine whole blood.ResultsThe ASA concentration in the synovial fluid (n = 6) reached a maximum of 4 μg/mL, the mean concentration over the entire perfusion period was 2 μg/mL. Maximum SA concentration was 17 μg/mL, the average was 14 μg/mL. ASA and SA concentration in the synovial fluid exceeded systemic concentrations 2 h and 3.5 h after “systemic” administration, respectively.ConclusionsASA and SA accumulated in the in the synovial fluid of the ex vivo model despite decreasing systemic concentrations. This suggests a prolonged anti-inflammatory effect within the joint that remains to be further elucidated.

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Section: Introductionmentioning

confidence: 99%

Synovial distribution of “systemically” administered acetylsalicylic acid in the isolated perfused equine distal limb

et al. 2013

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“…For horses, the study by Beaumier et al (1987) assessed that the maximum daily load of salicylate from their feed should not exceed 2 g given their feed composition (Keszycka et al, 2017). It should be noted that the digestive physiology of horses (monogastrics) is different from that of cattle (ruminants).…”

Section: Natural Sources Of Salicylic Acidmentioning

confidence: 99%

“…After this administration, salicylic acid was found only in negligible amounts in liver, muscle, plasma and eggs, with a maximum of 9.72 µg/kg in muscle and 82.3 µg/kg in eggs. Beaumier et al (1987) studied the presence of salicylic acid in the urine and plasma of Standardbred horses fed a twice-daily diet rich in alfalfa. Given the composition of the animal feed in this study, the maximum daily intake did not exceed 2 g of salicylates.…”

Section: Salicylic Acid Content Of Products Of Animal Originmentioning

confidence: 99%

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Salicylic acid residues in products of animal origin

Houben,

Delahaut,

Scippo

et al. 2024

EFS4

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“…Ζτςι, θ παρουςία των παραγϊγων του ςαλικυλικοφ οξζοσ ςτα άλογα μπορεί να ζχει ωσ πθγι προζλευςθσ ωυτικά είδθ που αποτελοφν μζροσ τθσ τροωισ τουσ ι/και ταυτόχρονα μπορεί να προζρχονται από εξωγενι χοριγθςι τουσ για κτθνιατρικι χριςθ και ςε αυτζσ τισ περιπτϊςεισ ζχουν αναωερκεί ιδιαίτερα υψθλζσ ςυγκεντρϊςεισ ςαλικυλικοφ οξζοσ ςτο αίμα και ςτα οφρα των αλόγων αυτϊν. [223][224][225] Ππωσ ζχει ιδθ αναωερκεί ςε προθγοφμενο κεωάλαιο, ςφμωωνα με το Άκρο 6 τθσ διεκνοφσ ςυμωωνίασ που ζχει εκδόςει θ Ραγκόςμια Ομοςπονδία των Αρχϊν Ιπποδρομιϊν (IFHA), ουςίεσ που μποροφν να προζρχονται από ωυτά που χρθςιμοποιοφνται παραδοςιακά για τθ βοςκι ι τθ ςυγκομιδι για τθν παραγωγι ηωοτροωϊν των αλόγων, όπωσ είναι το ςαλικυλικό οξφ, ςυμπεριλαμβάνονται ςτθν κατθγορία απαγορευμζνων ουςιϊν ορίου. Μετά από ςειρζσ μελετϊν που διεξιχκθςαν ςε πλθκυςμοφσ ειδϊν αλόγων για τον προςδιοριςμό των «ωυςιολογικϊν» ςυγκεντρϊςεων του ςαλικυλικοφ οξζοσ που ανιχνεφονται ωσ αποτζλεςμα πρόςλθψθσ μζςω τθσ τροωισ τουσ, ο AORC ζχει κεςπίςει ωσ ανϊτατο επιτρεπόμενο όριο ςυγκζντρωςθσ ςαλικυλικοφ οξζοσ ςτα οφρα και ςτο πλάςμα των αλόγων τα 750 μg mL -1 και 6,5 μg mL -1 , αντίςτοιχα.…”

Section: εισαγωγθ -σκορος τθς εγασιαςunclassified

Εφαρμογή Της Τεχνικής Της Υγροχρωματογραφίας / Φασματομετρίας Μαζών Με Αναλυτή Χρόνου Πτήσης (LC/TOF-MS) Στον Έλεγχο Ντόπινγκ Φαρμακευτικών Ουσιών Σε Ούρα Αθλητών Και Αλόγων

Βοναπάρτη¹

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In the current study, new qualitative and quantitative methods were developed andvalidated for the doping control screening analysis, confirmation, and/or quantitativedetermination of prohibited substances using LC/TOF-MS technology. The first part of thiswork presents the use of LC/TOF-MS for the multiple detection of a great number ofprohibited substances in athletes urine. In the second part, the same technology was appliedfor the quantification and identification of 4 threshold substances in horse urine using directinjection analysis.Unification of screening protocols for a wide range of doping agents has become animportant issue for doping control laboratories. In the present study, a high throughputscreening method was developed for the multiple detection of 266 small molecule analytesfrom all categories of prohibited substances included in the WADA List. The proposedmethodology is based on a single-step liquid-liquid extraction of hydrolyzed urine and theuse of a rapid-resolution LC/TOF-MS system acquiring continuous full scan spectral data.Electrospray ionization in the positive mode was applied. The extraction protocol andsample preparation procedure used in this study was applied in the OACA doping controllaboratory for the routine screening analysis of anabolic agents and corticosteroids. Ourapproach on sample preparation was to study the efficiency of this procedure for otherclasses of substances (stimulants, narcotics, diuretics). Validation parameters consisted ofidentification capability, limit of detection, specificity, ion suppression, extraction recovery,repeatability and mass accuracy. Detection criteria were established on the basis ofretention time reproducibility and mass accuracy. The suitability of the methodology fordoping control was demonstrated with positive urine samples. The preventive role of themethod was proved by the case where full scan acquisition with accurate massmeasurement allowed the retrospective reprocessing of acquired data from past dopingcontrol samples for the detection of a designer drug, the stimulant 4-methyl-2-hexanamine,which resulted in re-reporting a number of stored samples as positives for this particularsubstance, when, initially, they had been reported as negatives.Hydrocortisone is the primary endogenous glucocorticosteroid and is used in equineveterinary medicine for its anti-inflammatory properties. As an endogenous substance, itscontrol in equine sports is regulated by a threshold value of 1 μg ml-1 free hydrocortisone inurine. Two simple and rapid LC/MS methods with direct injection analysis were developedand validated for the quantification and identification of hydrocortisone in equine urineusing the same sample preparation but different mass spectrometric systems: IT-MS andTOF-MS. The main advantage of the proposed methodology is the minimal samplepreparation procedure, as particle-free diluted urine samples were directly injected intoboth LC/MS systems. Desonide was used as internal standard (IS). The tested linear rangewas 0,25–2,5 μg ml−1 for both methods. Matrix effects were evaluated by preparing andanalyzing calibration curves in water solutions and different horse urine samples. A greatvariation of the signal both for hydrocortisone and the internal standard was observed indifferent matrices. To overcome matrix effects, the unavailability of blank matrix and theexcessive cost of the isotopically labeled internal standard, standard additions calibration method was applied. This work is an exploration of the performance of the standardadditions approach in a method where neither non isotopic internal standards nor extensivesample preparation is utilized and no blank matrix is available. Validation results on linearity,accuracy and precision for both methods prove their suitability for the doping controlanalysis of hydrocortisone in horse urine. The above method was applied to a real sample,using both LC/MS methods.In equine sport, theobromine is prohibited with a threshold level of 2 μg mL-1 in urine. Twosimple LC/MS methods for the identification andquantification of theobromine weredeveloped and validated using the same sample preparation procedure but different massspectrometric systems (IT-MS and TOF-MS. Particle-free diluted urine samples were directlyinjected into the LC/MS systems, avoiding the time-consuming extraction step. 3-Propylxanthine was used as the internal standard. The tested linear range was 0.75–15 μgmL-1. Matrix effects were evaluated analyzing calibration curves in water and differentfortified horse urine samples. A great variation in the signal of theobromine and the internalstandard was observed in different matrices. To overcome matrix effects, a standardadditions calibration method was applied. The linearity, accuracy and precision for bothmethods prove their suitability for the doping control analysis of theobromine in horseurine. The methods were applied to two case samples, demonstrating simplicity, accuracyand selectivity.Salicylates have analgesic, anti-inflammatory, and antipyretic properties. In horses,salicylates may derive both from feeding as well as from medication. In equine sport,salicylic acid is prohibited with a threshold level of 750 μg mL−1 in urine. A simple and rapidLC/MS method was developed and validated for the quantification and identification ofsalicylic acid. Urine samples after 900-fold dilution and addition of the internal standard (4-methylsalicylic acid) were directly injected to the LC/QTOF-MS system. Electrosprayionization in negative mode with full scan acquisition mode and product ion scan mode werechosen for the quantification and identification of salicylic acid, respectively. Run time was2.0 min. The tested linear range was 2.5–50 μg mL−1 (after 100-fold sample dilution). Thelinearity, accuracy, and precision results prove the method’s suitability for the dopingcontrol analysis of salicylic acid in horse urine. The area ratios of the diagnostic product ionsof salicylic acid were found to be precise for confirmation purposes. The above method wasapplied to the quantitative and confirmatory analysis of two real samples.The endogenous catecholamine dopamine and its pharmacologically inert precursor, L-dopa,are both available as human pharmaceutical preparations and can be used as performanceenhancing drugs in horses. Urinary 3-methoxytyramine is used as an indicator ofdopaminergic manipulation resulting from dopamine or levodopa administration and isprohibited with a urinary threshold of 4 μg mL−1 (free and conjugated). A simple LC/MSmethod was developed and validated for the quantification and identification of 3-methoxytyramine in equine urine. Sample preparation involved enzymatic hydrolysis andprotein precipitation. ΗILIC was selected as a separation technique that allows effectiveretention of polar substances like 3-methoxytyramine and efficient separation from matrixcompounds. Electrospray ionization (ESI) in positive mode with product ion scan mode waschosen for the detection of the analytes. Studies on matrix effects showed ion suppressiondepending on the horse urine used. To overcome the variability of the results originatingfrom the matrix effects, isotopic labelled internal standard was used and linear regression calibration methodology was applied for the quantitative determination of the analyte. Thelinearity, accuracy and precision results prove the method’s suitability for the doping controlanalysis of 3-methoxytyramine in horse urine. The above method was applied to thequantitative and confirmatory analysis of three case samples.

show abstract

Presence of salicylic acid in Standardbred horse urine and plasma after various feed and drug administrations

Cited by 8 publications

References 7 publications

Synovial distribution of “systemically” administered acetylsalicylic acid in the isolated perfused equine distal limb

Synovial distribution of “systemically” administered acetylsalicylic acid in the isolated perfused equine distal limb

Salicylic acid residues in products of animal origin

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