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1.
An investigation was undertaken to demonstrate whether therapeutic treatment with ACTH raises hydrocortisone (cortisol) levels in horse urine above the limit (1000 ng/ml) established by the International Conference of Racing Authorities with the aim of controlling the abuse of cortisol and ACTH in equine sports. ACTH (200 iu) was administered i.m. to 3 Thoroughbred horses; urine and blood samples were collected at intervals afterwards and analysed by an immunoenzymatic system (ELISA) and HPLC-MS. To ascertain post exercise cortisol levels in untreated horses, 101 urine and 103 serum samples were taken from horses immediately after racing and analysed by ELISA. The peak urine level of cortisol, detected 8 h after ACTH administration, was around 600 ng/ml using either ELISA or HPLC-MS. The peak serum cortisol concentration was found to be around 250 ng/ml by ELISA, but consistently less by HPLC-MS. Mean cortisol levels in post race horses were 135.1+/-72.1 ng/ml in urine and 90.1+/-41.7 ng/ml in serum. High levels of the metabolite 20beta-dihydrocortisol in urine and the cortisol precursor 11beta-desoxycortisol in serum were found. The latter showed high cross-reactivity with cortisol on ELISA. In our experiment, treatment with ACTH 200 iu i.m. did not raise urinary cortisol levels above the 1000 ng/ml threshold proposed by the ICRA.  相似文献   

2.
Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.  相似文献   

3.
A survey of plasma and urinary concentrations of phenylbutazone and its metabolites in thoroughbred horses racing in Kentucky was carried out. Post-race blood samples from more than 200 horses running at Latonia Racetrack and Keeneland in the Spring of 1983 were analysed. The modal plasma concentration of phenylbutazone was between 1 and 2 micrograms/ml, the mean concentration was 3.5 micrograms/ml and the range was up to 15 micrograms/ml. Oxyphenbutazone had a modal plasma concentration between 1 and 2 micrograms/ml, a mean concentration of 2.07 micrograms/ml and a range of up to 13 micrograms/ml. gamma OH-phenylbutazone had a modal plasma concentration of less than 1 microgram/ml, a mean level of 1.39 micrograms/ml and a range of up to 7.32 micrograms/ml. All plasma concentration frequency distributions were well fitted by log normal distributions. Urinary concentrations of phenylbutazone yielded modal concentrations of less than 1 microgram/ml, a mean urinary concentration of 2.9 micrograms/ml, with a range of up to 30.5 micrograms/ml. This population fitted a log-normal distribution. For oxyphenbutazone the modal concentration was less than 3 micrograms/ml, the mean concentration was 15.26 micrograms/ml, with a range to 81.5 micrograms/ml. The frequency distribution of these samples was apparently bimodal. For gamma OH-phenylbutazone, the modal concentration was less than 4 micrograms/ml, the mean concentration 21.23 micrograms/ml, with a range of up to 122 micrograms/ml. The population frequency distribution for gamma OH-phenylbutazone was indeterminate. Analysis of the pH of these post-race urine samples showed a bimodal frequency distribution. The pH values observed ranged from 4.9 to 8.7, with peaks at about pH 5.25 and 7.25. This bimodal pattern of urinary pH values is consistent with observations made in England and Japan. Urinary pH influenced the concentrations of phenylbutazone, oxyphenbutazone and gamma OH-phenylbutazone found in the urine samples. The concentration of these metabolites found in alkaline urines were from 32 to 225 times greater than those found in acidic urines. Plasma concentrations of phenylbutazone and its metabolites, however, were unaffected by urinary pH. In interlaboratory experiments, horses running at Hollywood Park were dosed with phenylbutazone at about 2 g/1000 lbs 24 and 48 h before racing, and a mean dose of 0.6 g/1000 lbs at 72 h prior to racing. Post-race plasma samples from these horses showed phenylbutazone concentrations ranging from 0.44 to 9.97 micrograms/ml, with a mean concentration of 4.09 micrograms/ml.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

4.
Blood samples from the jugular vein and muscle biopsies (gluteus medius) in 25 Standardbred trotters were obtained 5-10 min after racing. The biopsies were analysed for fibre type composition and enzymatic profile and blood samples for plasma lactate and ammonia concentrations. Muscle characteristics, plasma lactate and ammonia concentrations after racing were compared with each horse's individual performance index (IPI). The IPI is calculated annually from the individual horse's racing performance (% placing 1, 2 or 3, total annual earnings, average earning per start, and best racing record), respectively, converted to and expressed as a percentage deviation from the average record of the same sex and age group. The IPI values were 100-116. Plasma lactate concentrations were 15.0-42.7 mmol/l (mean 31.3 mmol/l) and ammonia concentrations 65-210 micromol/l (mean 141 micromol/l) after racing. Fibre type composition varied among horses (range 9-27% for Type I, 32-54% for Type IIA, and 27-46% for Type IIB). Fibre type composition, enzyme activities, plasma lactate and ammonia responses to racing were not correlated to IPI. Ten of the horses also performed a submaximal test on the track, consisting of 5 incremental 1000 m heats at approximate speeds of 9.1, 9.5, 10.0, 10.5, and 11.1 m/s. Immediately after each heat a blood sample was drawn from the jugular vein for plasma lactate analysis. Plasma lactate response to exercise differed between horses, but no correlation was seen with IPI. Muscle characteristics, plasma lactate and ammonia concentrations after racing and lactate response to a submaximal track test did not correlate with current race performance expressed as IPI in a group of horses with average or better performance capacity at the time of testing. Analysis of lactate and ammonia in blood after racing is not a valuable tool to predict an individual performance index.  相似文献   

5.
A screening and confirmation procedure for drugs and metabolites in the blood serum and urine of racing animals was developed. Equine blood serum was spiked with low concentrations of several drugs of interest. Canine blood serum and urine were collected following oral doses of diethylcarbamazine, procaine, and phenylbutazone. Serum, urine, and extracts of each were analyzed, using a triple quadrupole mass spectrometer. Simultaneous screening of up to 50 drugs was possible in a single sample, in less than 2 minutes. Detection limits for most compounds were in the ng/ml to microgram/ml range, using 1-microliter samples. This procedure provided fast, sensitive screening for selected drugs and metabolites in blood serum and urine.  相似文献   

6.
Small volumes (0.05 ml) of cisterna magna cerebrospinal fluid (CSF) from 23 neurologically normal horses were analysed for the monoamine neurotransmitters dopamine, norepinephrine, epinephrine, serotonin and their metabolites using high pressure liquid chromatography and electrochemical detection. Two metabolites, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) were present in all CSF samples. The deaminated and methylated metabolite of dopamine, HVA, was present at a mean concentration of 42.33±3.14 ng/ml of CSF. The deaminated metabolite of serotonin, 5-HIAA, was present at a mean concentration of 45.52±3.65 ng/ml of CSF. A positive correlation was found between the CSF concentrations of HVA and 5-HIAA (r=0.72, p=0.0003). The mean ratio of HVA to 5-HIAA concentrations was 1.07±0.05. The ratios of HVA to 5-HIAA concentrations were found to be more indicative of the serotonergic metabolite 5-HIAA (r=–0.62, p=0.0016) than the dopaminergic metabolite HVA (r=0.11, p=0.60). There was a stronger relationship between the concentrations of 5-HIAA and the ratios of HVA to 5-HIAA in male CSF (r=–0.85, p=0.0006) than in female CSF (r=–0.54, p=0.01).Concentrations of CSF neurotransmitter metabolites were evaluated in horses grouped by age (0–4, 5–9 and 10–13 years). The youngest group of horses had a significantly (p=0.003) greater CSF concentration of HVA than of 5-HIAA. The 10–13-year-old horses had a significantly (p=0.02) lower mean CSF HVA concentration than the 0–4-year-old horses. No age-related differences in CSF 5-HIAA concentrations were detected.  相似文献   

7.
OBJECTIVE: To determine concentrations of dexamethasone in serum and urine of horses treated repeatedly with a topically administered ophthalmic dexamethasone preparation. ANIMALS: 4 clinically normal horses (2 mares, 2 geldings). PROCEDURE: 0.1% dexamethasone ophthalmic ointment was administered to the left eye of each horse every 5 to 9 hours for 8 consecutive days, yielding an estimated cumulative dexamethasone dose of 6.4 microg/kg of body weight. Serum and urine samples were obtained before the first dexamethasone treatment, on days 4 and 8 of treatment, and 24, 48, and 96 hours after cessation of treatment. To detect small concentrations of dexamethasone, serum and urine samples were analyzed by use of a competitive enzyme immunoassay. RESULTS: During the period of continued topical treatment, serum dexamethasone concentrations increased to between 0.10 and 0.49 ng/ml, then decreased below the limit of detection (0.06 ng/ml) within 24 hours after cessation of treatment. Dexamethasone also was detected in urine samples at concentrations of up to 0.98 ng/ml. CONCLUSIONS: Repeated topical administration of dexamethasone ophthalmic ointment generated low, but detectable glucocorticoid concentrations in serum and urine. CLINICAL RELEVANCE: Because treatment of performance horses with dexamethasone is prohibited for most types of competitions and because enhanced glucocorticoid detection methods may result in positive test results, owners and trainers may wish to reconsider entering horses in competitions during periods of treatment with ophthalmic dexamethasone preparations.  相似文献   

8.
Selegiline ([R]-[-]N,alpha-dimethyl-N-2- propynylphenethylamine or l-deprenyl), an irreversible inhibitor of monoamine oxidase, is a classic antidyskinetic and antiparkinsonian agent widely used in human medicine both as monotherapy and as an adjunct to levodopa therapy. Selegiline is classified by the Association of Racing Commissioners International (ARCI) as a class 2 agent, and is considered to have high abuse potential in racing horses. A highly sensitive LC/MS/MS quantitative analytical method has been developed for selegiline and its potential metabolites amphetamine and methamphetamine using commercially available deuterated analogs of these compounds as internal standards. After administering 40 mg of selegiline orally to two horses, relatively low (<60 ng/ml) concentrations of parent selegiline, amphetamine, and methamphetamine were recovered in urine samples. However, relatively high urinary concentrations of another selegiline metabolite were found, tentatively identified as N- desmethylselegiline. This metabolite was synthesized and found to be indistinguishable from the new metabolite recovered from horse urine, thereby confirming the chemical identity of the equine metabolite. Additionally, analysis of urine samples from four horses dosed with 50 mg of selegiline confirmed that N-desmethylselegiline is the major urinary metabolite of selegiline in horses. In related behavior studies, p.o. and i.v. administration of 30 mg of selegiline produced no significant changes in either locomotor activities or heart rates.  相似文献   

9.
A survey of standardbred horses was conducted to build up a normal population profile for insulin like growth factor-I (IGF-I) concentrations in racing standardbreds and to ascertain how age, sex and geographic location affect IGF-I. Blood samples were drawn by jugular venepuncture from 202 racing standardbred horses aged one to eight years located in five different geographic regions of New Zealand. IGF-I concentrations were determined by insulin like growth factor-I binding protein (IGFBP)-blocked radioimmunoassay validated for the horse. As described in other species, age played a significant (P<0.05) role in IGF-I concentrations with the highest concentrations occurring in the younger horses. There was a significant (P<0.05) sex effect, intact males having significantly higher IGF-I concentrations compared of mares and/or geldings. Geographic location had a significant (P<0.05) influence on IGF-I. A significant (P<0.05) trainer effect also was noted both within and between geographic locations. We concluded that IGF-I concentrations in racing standardbred horses are affected by age, sex, trainer and geographic location.  相似文献   

10.
Methylprednisolone acetate (MPA) is commonly administered to performance horses, and therefore, establishing appropriate withdrawal times prior to performance is critical. The objectives of this study were to describe the plasma pharmacokinetics of MPA and time‐related urine and synovial fluid concentrations following intra‐articular administration to sixteen racing fit adult Thoroughbred horses. Horses received a single intra‐articular administration of MPA (100 mg). Blood, urine, and synovial fluid samples were collected prior to and at various times up to 77 days postdrug administration and analyzed using tandem liquid chromatography‐mass spectrometry (LC‐MS/MS). Maximum measured plasma MPA concentrations were 6.06 ± 1.57 at 0.271 days (6.5 h; range: 5.0–7.92 h) and 6.27 ± 1.29 ng/mL at 0.276 days (6.6 h; range: 4.03–12.0 h) for horses that had synovial fluid collected (group 1) and those that did not (group 2), respectively. The plasma terminal half‐life was 1.33 ± 0.80 and 0.843 ± 0.414 days for groups 1 and 2, respectively. MPA was undetectable by day 6.25 ± 2.12 (group 1) and 4.81 ± 2.56 (group 2) in plasma and day 17 (group 1) and 14 (group 2) in urine. MPA concentrations in synovial fluid remained above the limit of detection (LOD) for up to 77 days following intra‐articular administration, suggesting that plasma and urine concentrations are not a good indicator of synovial fluid concentrations.  相似文献   

11.
Guanabenz (2,6-dichlorobenzylidene-amino-guanidine) is a centrally acting antihypertensive drug whose mechanism of action is via alpha2 adrenoceptors or, more likely, imidazoline receptors. Guanabenz is marketed as an antihypertensive agent in human medicine (Wytensin tablets, Wyeth Pharmaceuticals). Guanabenz has reportedly been administered to racing horses and is classified by the Association of Racing Commissioners International as a class 3 foreign substance. As such, its identification in a postrace sample may result in significant sanctions against the trainer of the horse. The present study examined liquid chromatographic/tandem quadrupole mass spectrometric (LC-MS/MS) detection of guanabenz in serum samples from horses treated with guanabenz by rapid i.v. injection at 0.04 and 0.2 mg/kg. Using a method adapted from previous work with clenbuterol, the parent compound was detected in serum with an apparent limit of detection of approximately 0.03 ng/ml and the limit of quantitation was 0.2 ng/ml. Serum concentrations of guanabenz peaked at approximately 100 ng/ml after the 0.2 mg/kg dose, and the parent compound was detected for up to 8 hours after the 0.04 mg/kg dose. Urine samples tested after administration of guanabenz at these dosages yielded evidence of at least one glucuronide metabolite, with the glucuronide ring apparently linked to a ring hydroxyl group or a guanidinium hydroxylamine. The LC-MS/MS results presented here form the basis of a confirmatory test for guanabenz in racing horses.  相似文献   

12.
The distribution of specific gravity values for 2,599 urine samples collected from racing Thoroughbred horses that were known to have received furosemide prior to racing was compared with that for 1,669 urine samples from racing Thoroughbred horses that reportedly had not received furosemide. Values of specific gravity for furosemide-treated horses were significantly lower (P < 0.001) than those for horses that had not received furosemide, and the proportion of horses with urine specific gravity either <1.010 or <1.012 was significantly greater (P < 0.001) among the furosemide-treated horses. These data indicate that evaluation of urine specific gravity would be a useful component of drug testing programs for regulation of furosemide use.  相似文献   

13.
Preparations of the herb, goldenseal, have been used in an attempt to thwart detection of morphine in urine of human addicts and racing horses. To assess the potential of goldenseal to interfere with equine drug detection, horses were dosed with morphine (0.1 mg/kg IV) and, in one experimental series, also with goldenseal (60 mg/kg, p.o.). Goldenseal resulted in significant increases in urine volume, specific gravity and acidity. A significant decrease in total morphine excreted occurred 5–6 hours after dosing. It was concluded that with inclusion of an efficient hydrolysis step in the analysis, goldenseal was unlikely to interfere with urine tests for morphine, even if water loading was included in the dosing regimen.  相似文献   

14.
A survey of the concentrations of cortisol in blood and urine samples taken from thoroughbred and standardbred horses after racing is presented. Statistical analysis showed the only significant difference between thoroughbred and standardbred horses was a higher cortisol concentration in thoroughbred urine. Urine volume and pH had no significant influence on the urinary cortisol concentration, however 9.5% of the urinary cortisol variation could be explained due to the influence of plasma cortisol concentration. The results of cortisol and ACTH administrations are also shown and compared with the survey results.  相似文献   

15.
Monitoring furosemide in racehorses participating in an EIPH program   总被引:1,自引:0,他引:1  
Analytical procedures were developed to monitor furosemide concentrations in post-race serum and urine samples obtained from horses participating in an exercise-induced pulmonary haemorrhage (EIPH) program. High performance liquid chromatography with ultraviolet light detection proved a reliable, sensitive method for measuring urinary furosemide concentrations up to 12 h after administration of either 150 or 250 mg of the drug to race horses. However, this method was unreliable for determination of serum furosemide concentration. High performance liquid chromatography with fluorescence detection proved a reliable, sensitive method for measuring serum furosemide concentrations in horses administered 250 mg of the diuretic, permitting detection approximately 5–10 ng/ml 6 h after treatment. This method was applied field conditions where furosemide was administered to horses (between 150 and 250 mg intravenously) 4 h prior to the race. Analytical results assisted establishing a threshold concentration of 85 ng/ml for serum furosemide. was found that serum furosemide concentrations are a valid measure of compliance with furosemide administration in the EIPH program.  相似文献   

16.
Medication control in greyhound racing requires information from administration studies that measure drug levels in the urine as well as plasma, with time points that extend into the terminal phase of excretion. To characterize the plasma and the urinary pharmacokinetics of flunixin and enable regulatory advice for greyhound racing in respect of both medication and residue control limits, flunixin meglumine was administered intravenously on one occasion to six different greyhounds at the label dose of 1 mg/kg and the levels of flunixin were measured in plasma for up to 96 hr and in urine for up to 120 hr. Using the standard methodology for medication control, the irrelevant plasma concentration was determined as 1 ng/ml and the irrelevant urine concentration was determined as 30 ng/ml. This information can be used by regulators to determine a screening limit, detection time and a residue limit. The greyhounds with the highest average urine pH had far greater flunixin exposure compared with the greyhounds that had the lowest. This is entirely consistent with the extent of ionization predicted by the Henderson–Hasselbalch equation. This variability in the urine pharmacokinetics reduces with time, and at 72 hr postadministration, in the terminal phase, the variability in urine and plasma flunixin concentrations are similar and should not affect medication control.  相似文献   

17.
The tricyclic antidepressant doxepin, representing a 5:1 mixture of trans- and cis-isomers, owns tranquilizing properties. This compound has been associated with illicit medication of racing horses, and therefore should be considered in doping control. Because analysis of doxepin in equine body fluids has not been documented in the literature, a highly sensitive analytical method was developed to individually monitor the doxepin isomers in blood and urine of horses by the use of gas chromatography/mass spectrometry. Following a dose of 1 mg doxepin-HCl/kg intravenously (i.v.), both the isomers were quantified for up to 24 h in serum of horses (n=4). The beta-half-lives of the trans- and cis-isomers were 3.5 and 3.1 h, respectively. The ratio of the trans/cis-isomers was found to be constant (4.7:1) during drug elimination and thus corresponded to the original composition of the antidepressant. Up to 12 h following administration low trans-isomer concentrations in an average range of 2-6 ng/mL were detected in urine of each of the horses, while the cis-isomer was only present in two of four horses for up to 8 and 12 h, respectively. In serum, mean trans-isomer concentrations exceeded urine levels maximally 120-fold after 3 h and at least sixfold after 12 h. As serum exhibits considerably higher concentrations of the doxepin isomers as compared with urine, blood of horses is the recommended body fluid when screening for the antidepressant.  相似文献   

18.
OBJECTIVE: To evaluate changes in plasma ionized calcium (Ca2+) and parathyroid hormone (PTH) concentrations in horses competing in endurance rides. DESIGN: Longitudinal clinical study. ANIMALS: 28 horses. PROCEDURE: Venous blood samples were obtained from horses before and after racing 80 km. Plasma pH and concentrations of Ca2+, PTH, inorganic phosphorus, albumin, lactate, and magnesium were measured. RESULTS: Overall, a significant decrease in mean (+/- SD) plasma Ca2+ concentration (from 6.44 +/- 0.42 to 5.64 +/- 0.42 mg/dl) and a significant increase in plasma PTH concentration (from 49.9 +/- 30.1 to 148.1 +/- 183.0 pg/ml) were found after exercise. Exercise also resulted in significant increases in plasma inorganic phosphorus, albumin, and lactate concentrations. No changes in plasma magnesium concentration or pH were detected after exercise. Plasma PTH concentration was not increased after exercise in 8 horses; in these horses, plasma PTH concentration decreased from 58.2 +/- 26.3 to 27.4 +/- 22.4 pg/ml, although plasma Ca2+ concentration was also decreased. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma Ca2+ concentration was decreased after racing for 80 km, compared with values obtained before racing. In most horses, an increase in plasma PTH concentration that was commensurate with the decrease in plasma Ca2+ was detected; however, some horses had decreased plasma PTH concentrations.  相似文献   

19.
Although endoscopy is the definitive diagnostic method for the detection of colonic ulcers, the equipment required for performing the test is costly and difficult to use. Therefore, a simple cost-effective and reliable screening test for intestinal tract bleeding is needed. To this end, we measured carbonic anhydrase isozymes (CA-I and CA-II) originating from erythrocytes by ELISA in order to determine if they could be used as markers of occult blood in feces. For fecal extract preparation, 2 g of feces were mixed with 4 ml of 0.01 M Tris-HCl (pH 8.0) containing 0.01% thimerosal. The concentrations of CA-I and CA-II in the fecal samples of 13 clinically normal racehorses were found to be 30.0 ± 10.0 and 34.0 ± 13.0 ng/ml, respectively. Increased concentrations of CA-I were detected in the fecal samples of 5 horses after blood administration; however, no increase was observed in CA-II. The concentrations of CA-I and CA-II in the fecal samples of 88 racehorses with clinical signs of equine gastric ulcer syndrome (EGUS) were 115.3 ± 79.0 and 41.0 ± 42.0 ng/ml, respectively. Thus, our results indicate that CA isozymes can be useful as markers of occult blood in the fecal samples of horses with intestinal tract bleeding.  相似文献   

20.
Horse racing is a contest between horses, usually held for the purpose of betting. Thoroughbred horse racing is the most diffused form of horse racing throughout the world. Thoroughbred is one of the most versatile of horse breeds and has influenced the development of many other breeds. Thoroughbred horses served as a foundation stock for the development of the light horse breeds. The two types of horse racing are flat racing and jumping races/steeplechases. The measures of racing performance are broadly classified into three categories. They are time and its several variations, handicap or similar performance ratings and earnings. One common measure of the performance of racehorses evaluated genetically is racing time or final time. The heritability estimates differed according to method of estimation, age, sex, track and distance. Time measure generally had a heritability in the range of 0.1 to 0.2 with the higher values for shorter races. For handicap and earning measures the heritabilities reported were generally higher in the range of 0.3 to 0.4; hence these may be considered in genetic evaluation of racing performance of Thoroughbred horses. The average generation interval of Thoroughbred horses was 11.2 ± 4.5 and 9.7 ± 3.8 years for males and females respectively, which limits the genetic progress in racing horses. However, the major advantage is that the racing performance may be evaluated in both males and females and repeated observations can be obtained on the same animal in relatively short periods. These factors coupled with the reasonable heritability of some measures of racing performance, suggest that mass selection based on performance tests would be the selection procedure of choice to improve the racing performance of Thoroughbred horses. In general, the inbreeding at the rate that is usually practised in Thoroughbred population does not enable much gene fixing. However, practice of close inbreeding may be avoided, even though it still fascinates breeders at subconscious level.  相似文献   

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