Urine analysis in horses involves the examination of urine to assess the health and physiological status of the animal. This diagnostic tool provides insights into the renal function, hydration status, and potential presence of systemic diseases. Key parameters evaluated in equine urine analysis include pH, specific gravity, protein levels, glucose, ketones, and the presence of blood or sediment. Variations in these parameters can indicate metabolic disorders, infections, or other health issues. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, findings, and clinical implications of urine analysis in equine veterinary practice.
Trim CM, Hanson RR.The intravenous administration of xylazine (1.1 mg/kg bodyweight) in six ponies resulted in a significant increase in urine output over two hours, with maximum flow occurring between 30 and 60 minutes after injection. Urine specific gravity, osmolality and glucose concentration decreased. Renal clearance of endogenous creatinine was unchanged. Significant increases in the excretion of potassium and chloride occurred. Plasma glucose concentration was increased 30 minutes after the administration of xylazine by a mean value of 37 per cent. Serum osmolality and sodium, potassium and chloride conc...
Johansson IM, Anlér EL, Bondesson U, Schubert B.Two metabolites of meclofenamic acid have been isolated from equine urine. Both metabolites are found to be monohydroxylated forms of meclofenamic acid by gas chromatography-mass spectrometry after extractive alkylation. The parent drug and the metabolites are separated by reversed-phase liquid chromatography on a Spherisorb ODS column, using methanol-phosphate buffer eluents and UV detection at 280 nm. The structure of the metabolites is discussed on the basis of LC, TLC and GC-MS data.
Skrabalak DS, Henion JD.Betamethasone and its major unconjugated metabolite, 6-beta-hydroxybetamethasone, were detected in equine urine by thin-layer chromatography and characterized by micro-liquid chromatography/mass spectrometry (micro-LC/MS). Their structures were confirmed by a combination of infrared spectroscopy and nuclear magnetic resonance spectroscopy.
Her GR, Watson JT.Sensitive and specific methodology based on capillary column gas chromatography-electron capture-negative chemical ionization-mass spectrometry has been developed for the quantitative analysis of corticosteroids from biological fluids. The feasibility of this method is demonstrated in the quantitative analysis of dexamethasone in horse urine following administration of the drug. A structurally similar compound, 6 alpha-methylprednisolone, is added to the urine as an internal standard. The free dexamethasone and the internal standard are extracted and oxidized to high-electron-affinity 1,4-andr...
Gallicano KD, Ng RM, Young LM.Methylprednisolone and three metabolites, 17,21-dihydroxy-6 alpha-methyl-1,4-pregnadiene-3,11,20-trione, 6 alpha-methyl-17,20 beta,21-trihydroxy-1,4-pregnadiene-3,11-dione, and 6 alpha-methyl-11 beta,17,20 beta,21-tetrahydroxy-1,4-pregnadien-3-one were detected in equine urine after intraarticular administration of methylprednisolone acetate. All four compounds were excreted both in the unconjugated form and as glucuronic acid conjugates. They were identified by comparing data obtained from analyses by high performance liquid chromatography, thin-layer chromatography, ultraviolet spectroscopy ...
Gronwall R.Endogenous creatinine clearance and renal excretion of phenylbutazone, osmotically active material, and compounds contributing to the urinary refractive index were studied in 12 Thoroughbred mares after no treatment, after water administration, or after furosemide administration. Urine was quantitatively collected, using urinary bladder catheters. On average, urine flow of the mares was 9 microliters/min/kg without treatment and increased to about 50 microliters/min/kg after water administration and to about 70 microliters/min/kg after furosemide administration. Water administration increased ...
Skrabalak DS, Cuddy KK, Henion JD.Micro-liquid chromatography-mass spectrometry (micro-LC-MS) was utilized to quantitatively determine betamethasone and its major unconjugated metabolite, 6 beta-hydroxybetamethasone, in equine plasma and urine. The advantage of micro-LC-MS over conventional gas chromatography-mass spectrometry in corticosteroid determination is illustrated and the reliable, steadfast nature of micro-LC-MS is demonstrated through example.
Houston T, Chay S, Woods WE, Combs G, Kamerling S, Blake JW, Edmundson AG, Vessiney R, Tobin T.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 microgram...
Haddad NS, Pedersoli WM, Ravis WR, Fazeli MH, Carson RL.Gentamicin (GT) was administered IM to 6 healthy mature mare ponies at a dosage of 5 mg/kg of body weight every 8 hours for 7 consecutive days (total, 21 doses). Two venous blood samples were collected before (trough) and at 1 hour (peak) after the 5th, 10th, 14th, and 19th doses. An endometrial biopsy was done of each mare on days 4 and 7. On the 7th day, just before the 21st administration of GT, base-line blood samples were collected, and 22 blood samples were collected over a period of 48 hours after GT was given. The mares were catheterized on the 7th day, and urine was collected for 24 h...
Woods WE, Chay S, Houston T, Blake JW, Tobin T.Interference or 'masking' in thin layer chromatography occurs when the presence of one drug on a thin layer plate physically obscures or interferes with the detection of another drug. We investigated the ability of phenylbutazone and oxyphenbutazone to mask or interfere with the detection by high performance thin layer chromatography (HPTLC) of basic drugs used illegally in horse racing. Of fifty-five basic drugs called 'positive' since 1981 by laboratories affiliated with the Association of Official Racing Chemists (AORC), forty did not comigrate with phenylbutazone or oxyphenbutazone and cou...
Gronwall R, Price G.To determine the rate of urine flow and thus urinary excretion in the horse from untimed urine samples alone, the flow rate, creatinine concentration, osmolarity, and refractive index of 228 quantitatively collected urine samples were determined in 53 experiments on 12 healthy Thoroughbred mares. Forty samples were collected after water-induced diuresis; 11 samples were collected after furosemide-induced diuresis. Flow rates, which ranged from 1.2 to 84.5 ml/min, could be predicted from the urinary creatinine concentration. Correlation of urinary flow with urinary creatinine concentration acco...
Adams R, McClure JJ, Gossett KA, Koonce KL, Ezigbo C.gamma-Glutamyltranspeptidase (GGT) activity in equine urine was measured, using an assay developed for use with serum and was found to be reproducible. The GGT activity was measured in samples prepared by serial dilution of exogenous GGT with equine urine, and the activity was determined to be linear between 21 IU/L and 407 IU/L. The behavior of exogenously added GGT was compared in equine serum and urine. The enzyme behaved similarly in both fluids. The GGT activity was measured in serum and urine samples after storage at -20, 4, and 25 C for 24 and/or 72 hours. Enzyme activity decreased afte...
Gallicano KD, Park HC, Young LM.A sensitive method was required to analyze low levels of camphor in equine urine and plasma. Camphorated oil (20% w/w camphor) was administered topically (6 g) and intratracheally (1 g) to standardbred mares. The drug was extracted from urine and plasma by diethyl ether and analyzed as its 2,4-dinitrophenylhydrazone derivative by reverse phase HPLC with UV detection. The UV detector was set at 368.5 nm and the samples were eluted from the C18 column by 82% acetonitrile in water. The detection limit achieved was about 10 ng/mL urine and about 20 ng/mL plasma. After topical administration, only ...
Skrabalak DS, Covey TR, Henion JD.Several important corticosteroids were qualitatively determined in the plasma and urine of horses by micro-liquid chromatography-mass spectrometry (micro-LC-MS). The sensitivity and specificity of micro-LC-MS are demonstrated as is the ability of micro-LC-MS to deal with endogenous interferences. In turn, the relative amount of dexamethasone and its major unconjugated metabolite were determined in equine urine by micro-LC-MS; the conclusions drawn are reported.
Brotherton HO, Yost RA.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, u...
Morris DD, Divers TJ, Whitlock RH.Four consecutive 6-hour urine collections were obtained from 10 healthy adult horses given free access to a complete pelleted ration and water. Samples of blood were collected from the jugular vein at the start of the test period and then every 6 hours for 4 consecutive samples. Mean renal clearance of creatinine (CCr) varied significantly during the four, 6-hour collections, ranging from 1.474 in period II to 2.702 ml/min/kg of body weight during period III (P less than 0.05). The CCr in period III was significantly different from the 24-hour mean value of 1.877 ml/min/kg (P less than 0.05). ...
Edwards KE, Stevens S, Woodward CB, Tweeten KA.Counterimmunoelectrophoresis was evaluated as a method to distinguish urine of human origin from that of equine origin. The procedure used anti-equine serum and anti-human serum antibodies that had been solid-phase absorbed to eliminate species cross-reactivity. Counterimmunoelectrophoresis reliably detected contamination of equine urine by human urine to a level of 10% with a minimum sensitivity to about 2% contamination. Compared with double diffusion, counterimmunoelectrophoresis was approximately 10 to 15 times more sensitive in the detection of urine proteins.
Soma LR, Korber K, Anderson T, Hopkins J.The effects of furosemide (0.55 mg/kg IV) on the plasma and urinary fentanyl (PFE UFE) concentrations were studied during steady-state conditions. The PFE during the steady-state period was 0.31 +/- 0.027 ng/ml, with no significant changes occurring, even though the rate of excretion of fentanyl (EX) increased during the 1st hour from 112.0 +/- 21.6 to 534.5 +/- 82.9 ng/minute. The EX returned to control levels within 3 hours, as did the UFE. The injection of furosemide increased glomerular filtration rate from 1.97 +/- 0.21 to 3.81 +/- 0.75 ml/kg/min. The fractional reabsorption decreased fro...
Brown MP, Embertson RM, Gronwall RR, Beal C, Mayhew IG, Curry SH.Six mares were given 5 IM injections (at 12-hour intervals between doses) of amikacin sulfate at a dosage of 7 mg/kg of body weight. Serum amikacin concentrations were measured serially throughout the study; synovial, peritoneal, endometrial, and urine concentrations were determined after the last injection. Amikacin concentrations of the CSF were measured serially in 3 of the 6 mares; 1 of the 3 mares had septic meningitis. Mean serum amikacin concentrations peaked at 1 to 2 hours after IM injection. The highest mean serum concentration was 19.2 micrograms/ml (1.5 hours after the 5th injectio...
Dumasia MC, Houghton E.The metabolism of 19-nor[4-14C]testosterone has been studied in the equine castrate. Following XAD-2 extraction of aliquots of the 0-24 h urine samples, the glucuronic acid and sulphate conjugates were separated by Sephadex LH-20 column chromatography. After hydrolysis of the conjugates, the neutral phase I metabolites of 19-nortestosterone were extracted, purified and identified by g.l.c.-mass spectrometry. In phase I metabolism stereospecificity was observed in the reduction of the A-ring with the formation of the 5 alpha, 3 beta-isomers of estranediol. Epimerization at C-17 and hydroxylatio...
Mount ME, Feldman BF.Strychnine toxicosis is characterized by inducible tetanic seizures and metaldehyde poisoning by fine fasciculations progressing to generalized tremors and seizures. Intoxication with 1080 causes seizures, random running movements, vomiting, defecation, urination, acidosis and hyperglycemia. Intoxication with rodenticides causing coagulopathy is characterized by hemorrhage into body cavities but not necessarily external hemorrhage. Anticholinesterase insecticides cause salivation, urination and defecation, while chlorinated hydrocarbon insecticides cause CNS disturbances. Ethylene glycol intox...
Seanor JW, Byars TD, Boutcher JK.Renal dysfunction secondary to GI disorders may be relatively common in horses. Persistent dehydration of 8-10% of body weight can lead to prerenal azotemia, which may result in renal ischemia and renal disease if uncorrected. Dehydrated azotemic horses with a urine specific gravity less than 1.018 may have renal disease. Urine specific gravity readings greater than 1.025 usually indicate normal kidney function. A urine Na level less than 20 mEq/L and a urine/plasma creatinine ratio greater than or equal to 20:1 indicate prerenal problems. Use of nephrotoxic drugs should be avoided in septicem...
Ketelaars HC, Peters JG, Anzion RB, Van Ginneken CA.The isolation, partial identification and quantitative determination of four guaiphenesin glucuronides in plasma and urine of the horse is described. The identity of the glucuronides was checked by UV and fluorescence spectrophotometry, by NMR spectrometry and by mass spectrometry after permethylation. The applicability of the procedure to pharmacokinetic studies is demonstrated.
Brown MP, Kelly RH, Gronwall RR, Stover SM.Six healthy adult mares were given a single IV dose (25 mg/kg of body weight) of chloramphenicol sodium succinate. Chloramphenicol concentrations in serum, synovial fluid, peritoneal fluid, and urine were measured serially over a 48-hour period. The highest measured serum chloramphenicol concentration was 6.21 micrograms/ml at 0.5 hour. Chloramphenicol was detected in synovial and peritoneal fluids, with mean peak concentrations of 3.89 micrograms/ml and 3.50 micrograms/ml, respectively, at 0.5 hour. Serum and synovial concentrations declined rapidly and were not measurable at 3 hours. Chloram...
Houghton E, Copsey J, Dumasia MC, Haywood PE, Moss MS, Teale P.As part of a continuing research program associated with the detection of anabolic steroid residues in horse urine, normal samples from entire male horses have now been investigated. Isomers of three C-18 neutral steroids; 4-estren-17-ol-3-one (1), estrane-3,17-diol (2) and an unsaturated estranediol having a possible structure (3), have been identified in urine samples from two male horses aged 8 and 14 years. Of these three steroids, compound (2) was not detected in the urine of a 2.5 yr old entire male nor in the majority of post-race urine samples from entire male horses average age 3.8 yr...
Thurmon JC, Steffey EP, Zinkl JG, Woliner M, Howland D.Xylazine given IV at doses of 0.5, 1.0, and 1.5 mg/kg to mares caused a significant (P less than 0.05) dose-related increase in serum glucose concentration and urine volume. Serum glucose concentrations as much as 150 mg/dl were recorded in mares after they were given the largest xylazine dose. The greatest urine volume, similar to changes in peak glucose concentration, always occurred during the first hour after dosing with xylazine and averaged 1.82, 3.93, and 5.68 ml/kg/hour after the 0.5-, 1.0-, and 1.5-mg/kg doses, respectively, were given. Urine osmolality and specific gravity were signi...
Dewey EA, Maylin GA.The metabolism of propionylpromazine in the horse was studied. Although propionylpromazine is not currently approved or recommended for use in horses, it has been used illegally to alter their performance. Propionylpromazine hydrochloride was administered intramuscularly at clinical and subclinical doses. Three metabolites were detected in urine. The major metabolite was identified as 2-(1-hydroxypropyl) promazine sulfoxide. The detection of this metabolite in routine drug testing has been described.
Moss M S..A review of positive Jockey Club "dope tests" during the 12 years from 1970 to 1981 inclusive is presented and a comparison made with certain overseas racing authorities for varying periods between 1975 and 1981. Urinary pH of post race urine samples is predominantly acidic (although varying in a significant manner throughout the year) and thus favours excretion of the generally more potent basic drugs. The proportion of positive results was about the same in winners and non-winners, ie, one in 400 horses sampled. Drugs found fell mainly into four categories: methylxanthines; non-steroidal ant...
Delbeke FT, Baert K, De Backer P.Urinary and plasma concentrations of the nonsteroidal anti-inflammatory drug tiaprofenic acid were determined following oral and intramuscular administration of a dose of 1 g to five fasted horses. Quantitation was performed by high-performance liquid chromatography (HPLC). The limit of quantitation (LOQ) was 0.1 microg/ml and 0.5 microg/ml in 2 ml plasma and 1 ml urine, respectively. Assay precision and extraction recovery were between acceptable values. Tiaprofenic acid pharmacokinetics were described by non-compartment analysis of the data. Absorption was faster after oral administration as...
Maxwell L, McCarrel TM, Hay S, Cole C.The antifibrinolytic agent aminocaproic acid (ACA) is occasionally used prior to episodes of intense training in racehorses suffering from exercise-induced pulmonary hemorrhage. Although a previous study indicated that the drug is cleared rapidly in horses, some racetrack practitioners claim that recent adverse analytical findings for ACA in postrace samples were from ACA administrations 5-7 days before the race. The purpose of this study was to re-examine the pharmacokinetics of ACA in horses to address this apparent conundrum. Eight exercise-conditioned thoroughbred horses were administere...
Soma LR, Uboh CE, Rudy JA, Smith MS.To examine, in horses, the disposition and excretion of the active metabolite 6-methoxy-2-naphthylacetic acid (6MNA) of the nonsteroidal anti-inflammatory prodrug nabumetone. Methods: Pharmacokinetic analysis of 6MNA after oral administration of nabumetone and IV administration of 6MNA. Methods: Using a crossover design, 5 horses were orally administered 3.7 mg of nabumetone/kg of body weight. After a 3-week washout period, 4 horses were administered 2.5 mg of 6MNA/kg, IV. Results: Absorption of nabumetone from the gastrointestinal tract and its metabolism to 6MNA had a median appearance half-...
Brobst DF, Lee HA, Spencer GR.An 11-year-old mare with polyuria, polydipsia, and azotemia was found to be hypercalcemic and hypophosphatemic. The concentration of calcium in a single collection of urine was within normal limits, although urinary inorganic phosphate concentration was lower than normal. After a brief period of supportive treatment, the mare died. At necropsy, the kidneys were found to be shrunken and fibrous. Histologically, the lesions were those of glomerulonephritis.
Rivas LJ, Hinchcliff KW, Kohn CW, Sams RA, Chew DJ.To describe changes in renal function of horses after oral and i.v. administration of sodium bicarbonate (NaHCO3) and to determine whether changes are dose dependent. Methods: 6 Standardbred mares. Methods: Blood and urine samples for determination of renal function were collected immediately before and at hourly intervals for 12 hours after administration of each of 3 oral doses (1,500, 1,000 and 250 mg/kg of body weight, in 3 L of water) and 1 i.v. dose (250 mg/kg, 5% solution) of NaHCO3, or water (3 L orally). Results: NaHCO3 induced increases in urine flow; electrolyte-free water reabsorpt...
Wood T, Tai CL, Taylor DG, Woods WE, Wang CJ, Houtz PK, Tai HH, Weckman TJ, Yang JM, Sturma L.Detomidine is a potent non-narcotic sedative agent which is currently in the process of being approved for veterinary clinical use in the United States. Since no effective screening method in horses is available for detomidine, we have developed an 125I radioimmunoassay for detomidine in equine blood and urine as part of a panel of tests for illegal drugs in performance horses. Our 125I radioimmunoassay has an I-50 for detomidine of approximately 2 ng/ml. Our assay shows limited cross-reactivity with the pharmacodynamically similar xylazine, but does not cross-react with acepromazine, epinephr...
Paine SW, Aldurdunji M, Garth-Greeves A, Muir T, Hincks PR.Hydroxyzine and cetirizine are first- and second-generation oral antihistamine drugs, respectively, used to treat allergic reactions in horses. Cetirizine is also a metabolite of hydroxyzine, which may lead to complexities in regulating their use in equine sporting events. The aim of the research was to be able to provide detection times (DT) from pharmacokinetic studies in thoroughbred horses to better inform trainers, and their veterinary surgeons, prescribing these substances for treatment of Thoroughbred racehorses. Six and two horses were given 9 repeated administrations of hydroxyzine HC...
Abd El Kader NA, Farghali HA, Abu-Seida AM, Salem NY, Khattab MS.Early detection of cystitis in equine is essential to improve the prognosis and outcome of therapy. However, the conventional white light endoscopy is not sufficiently accurate for this purpose. Hence, this study evaluated chromoendoscopy as a recent diagnostic tool for cystitis in female donkeys. For this purpose, 5 apparently normal donkeys (control group) and 5 female donkeys with cystitis (diseased group) were used. Physical and rectal examinations, urine analysis, white light cystoscopy, methylene blue-based chromoendoscopy and histopathology were performed in all animals. Turbid urine ex...
Schott HC, Patterson KS, Eberhart SW.To determine whether administration of glycerol-containing solutions induces a state of transient hyperhydration in resting euhydrated horses, changes in plasma and urine constituents were measured in four horses for 1 h before and 5 h after nasogastric administration of each of four treatments (Experiment 1). Treatments were applied in a randomized fashion and included: (1) 1.0 g.kg(-)(1)glycerol in 8 L of water (G); (2) 8 L of water (W); (3) 8 L of 0.9% NaCl solution (S); and (4) 1.0 g.kg(-)(1)glycerol in 8 L of 0.9% NaCl solution (GS). In a subsequent study, voluntary water intake was measu...
Mutoh K, Watanabe S, Wakuri H.The glands of pelvis renalis in adult and fetal horses were examined by histochemically. The glandular terminal was divided into two types, intraepithelial gland (IE) and extraepithelial gland (EE) by their locations and histochemical characters. Both glands were composed of mucous cells. The former lay in the transitional epithelium and were stained reddish with galactose oxidase-Schiff (GOS) and mild oxidation-Schiff (MOS), bluish purple with periodic acid-cold thionine Schiff-NaOH-PAS (PCP). The latter distributed in the lamina propria, but was not discovered in the fetus. They were stained...
Cartoni GP, Montanaro M.The results and the improvement of the analytical procedures adopted for the control of doping in horses will be reported. This control has been systematically carried out in Italy for about 10 years in the laboratories of Italian Federation of Sport and Medicine in which the biological samples for the control of doping in various sport activities (football, cycling, athletics etc.) are also examined. In this way it is possible to use the same instruments for all these similar problems and compare the results. The analytical procedure is based on the following steps: 1) Extraction of the sampl...
Chou CC, Chen CL, Asbury AC, Webb AI, Vickroy TW.To develop an ELISA that is sensitive and suitable for measurement of immunoreactive acepromazine (ACP) in horse serum and urine and to determine the acute effects of exercise on immunoreactive ACP values in Thoroughbreds. Methods: 12 healthy Thoroughbreds (5 mares, 5 geldings, 2 stallions), aged 2 to 8 years. Methods: A commercially available antibody and a horseradish peroxidase-conjugated oxime derivative of immunoreactive ACP were used to develop a one-step ELISA. Horses were used in a crossover design study to evaluate possible effects of treadmill exercise on serum and urine ACP concentr...
Van Eenoo P, Delbeke FT, Roels K, Baert K.Non-steroidal anti-inflammatory drugs (NSAIDs) are prohibited by the International Federation of Horse Racing Authorities but are commonly used in veterinary practice. Plasma and urinary concentrations of the NSAID tolmetin were determined by a high-performance liquid chromatographic procedure with UV detection following oral administration of a dose of 1 g to six fasted untrained standard bred mares. With a limit of quantitation (LOQ) of 0.05 microg/ml tolmetin was present in plasma for 9-12 h post-administration. Maximum concentrations of 2.1+/-0.89 microg/ml were found after 0.7+/-0.25 h. T...
Roberts CJ, Jackson LS.The development, validation, and application of an ELISA for dexamethasone in equine urine is described. The drug-protein conjugate was immobilised in microtitre plate wells and antiserum raised against the same drug-protein conjugate was allowed to compete with sample or standard drug and the immobilised drug-protein conjugate. The proportion of antiserum binding to the immobilised drug-protein conjugate was detected using a biotinylated protein G/extravidin-alkaline phosphatase complex in situ and measurement of the substrate product. The method was used to detect the presence of drug-derive...
Black SB, Stenhouse AM, Hansson RC.This paper details various rapid and sensitive methods for the extraction and derivatisation of propranolol, metoprolol, sotalol, atenolol, pindolol, timolol, oxprenolol, alprenolol and penbutolol in equine urine and in human post mortem whole blood and urine. Three solid-phase extraction methods are described involving the use of either XtrackT XRDAH515, Bond Elut Certify or Sep-Pak C18 cartridges. Two derivatisation methods are also described involving the formation of cyclised silyl or pentafluoropropionate derivatives with either chloromethyldimethylchlorosilane or pentafluoropropionic anh...
Kim Y, Seo C, Oh S, Kwak J, Jung S, Sin E, Kim H, Ji M, Lee HS, Park HJ, Lee G, Yu J, Kim M, Lee W, Paik MJ.Loxoprofen is a non-steroidal anti-inflammatory drug of the 2-arylpropionic acid type, which has used to treat musculoskeletal disorders in the horse racing industry. However, it has also used illicitly to mask clinical signs of inflammation and pain in racehorses. Thus, its accurate analysis has become an important issue in horse doping laboratories. In this study, an analytical method of loxoprofen was developed as tert-butyldimethylsilyl (TBDMS) derivative by gas chromatography-mass spectrometry (GC-MS). Characteristic fragment ions of [M-15], [M-57], and [M-139] permitted the accurate and ...
Delbeke FT, Landuyt J, Debackere M.A high-performance liquid chromatographic method to measure plasma and urinary alclofenac levels in equine biofluids is described. Isolation of the drug from plasma is achieved using liquid-liquid extraction with diethyl ether. Reversed-phase C18 solid phase extraction is used for the extraction of free and conjugated alclofenac from urine. The reproducibility and accuracy of the method were well within acceptable limits over the concentration ranges 0-10 and 0-20 micrograms/ml, respectively, for plasma and urine. Starting with 2 ml of plasma, a concentration of 0.1 microgram/ml could easily b...
Hülsemann F, Flenker U, Machnik M, Schänzer W.Sodium bicarbonate administration to horses prior to competition in order to enhance the buffer capacity of the organism is considered as a doping offence. The analysis of the isotopic composition of urinary bicarbonate/CO(2) (TCO(2)) may help to identify an exogenous bicarbonate source, as technical sodium bicarbonate exhibits elevated delta(13)C values compared with urinary total carbon. The isolation of TCO(2) from 60 equine urine samples as BaCO(3) followed by an isotopic analysis shows a significant variability of delta(13)C for TCO(2) of more than 10 per thousand. The delta(13)C of total...
Bland SA, Blake JW, Ray RS.Mefenamic acid is extracted from biological fluids and is acylated with pentafluoropropionic anhydride to form a derivative possessing high electron affinity. The derivative is analyzed by gas-liquid chromatography with an electron capture detector. The method is particularly valuable for determining drug levels in blood where small sample and/or drug concentrations are available.
Ketelaars HC, Peters JG, Anzion RB, Van Ginneken CA.The isolation, partial identification and quantitative determination of four guaiphenesin glucuronides in plasma and urine of the horse is described. The identity of the glucuronides was checked by UV and fluorescence spectrophotometry, by NMR spectrometry and by mass spectrometry after permethylation. The applicability of the procedure to pharmacokinetic studies is demonstrated.
Leung DKK, Wong ASY, Zhou QL, Wan TSM, Ho ENM.A non-target variable Data Independent Acquisition (vDIA) workflow based on accurate mass measurements using a Q Exactive OrbiTrap is presented for the first time for equine doping control testing. The vDIA workflow uses a combination of MS1 events (1 to 2) and multiple vDIA events to cover the analytes of interest. The workflow basically captures a digital image of a sample allowing all relevant MS1 and MS2 data to be recorded. In theory, the workflow can accommodate an unlimited number of analytes as long as they are amenable to the sample extraction protocol and fall within the mass limits ...
Divers TJ, Byars TD, Murch O, Sigel CW.Proteus mirabilis cystitis was induced in 9 ponies by chemically eroding the bladder mucosa before the organism was inoculated. Comparisons were made in the treatment of P mirabilis cystitis between ponies treated daily for 13 days with a trimethoprim-sulfadiazine (TMP-SDZ) paste and both positive and negative controls. Urine cultures from ponies treated with TMP-SDZ became negative for P mirabilis between days 3 and 9 after the start of the treatment, whereas positive controls remained infected until day 13. Urine cultures from all ponies were negative for P mirabilis on day 28. Urine concent...
Skrabalak DS, Cuddy KK, Henion JD.Micro-liquid chromatography-mass spectrometry (micro-LC-MS) was utilized to quantitatively determine betamethasone and its major unconjugated metabolite, 6 beta-hydroxybetamethasone, in equine plasma and urine. The advantage of micro-LC-MS over conventional gas chromatography-mass spectrometry in corticosteroid determination is illustrated and the reliable, steadfast nature of micro-LC-MS is demonstrated through example.
Törneke K, Larsson CI, Appelgren LE.Isoxsuprine is used in veterinary medicine as a vasodilating agent. The drug has been detected in the urine of horses up to 6 weeks after the cessation of administration. In the present study, the distribution pattern of 3H-isoxsuprine was investigated using whole body autoradiography in mice to find a possible site of retention. Melanin was the only place of retention identified. Additional in vitro studies showed an affinity of isoxsuprine to both melanin and keratin. The K(d) values were 0.02 mmol/l and 1 mmol/l, and the B(max) values were 0.2 micromol/mg and 2 micromol/mg, respectively. A ...
Uboh CE, Soma LR, Rudy JA, Morgan E, Mengeringhausen K, Sams R.This study was undertaken to determine the applicability of plasma concentration of furosemide and specific gravity (SG) of urine in regulating the use of furosemide administered 4 hours prior to race time in Exercise-Induced Pulmonary Hemorrhage (EIPH) race horses. Nonbleeders (CTL) and certified bleeders (FUR) actively racing in Illinois (IL) and Pennsylvania (PA) were used in the study. Various doses (less than 250, 250, 300, 350, 400 and 500 mg) were administered either as a single intravenous (IV) dose or as a combination (IV-IM) of IV and intramuscular (IM) administrations 4 hours before...
Hunt JP, Haywood PE, Moss MS.A gas chromatographic screening procedure for the non-steroidal anti-inflammatory group of drugs is described. The method invovles on-column methylation of the carboxylic acid group using trimethylanilinium hydroxide as the methylating reagent. Fifteen such drugs were studied. Eight of these were detected in urine collected from horses that had received these compounds orally and for these drugs, rates of urinary excretion are recorded. Seven other members of this group of drugs were shown to be detectable by this procedure but in these cases the drug was added to urine and not administered to...
Singh AK, Mishra U, Ashraf M, Abdennebi EH, Granley K, Dombrovskis D, Hewetson D, Stowe CM.Chemical ionization- and electron impact ionization-selective ion monitoring provided a simple and sensitive method for measuring detomidine (Domosedan), a potent sedative-analgesic drug for horses and cattle. Chemical ionization was at least 10 times more sensitive than electron impact ionization. By using propranolol as an internal standard, we found that the recovery of detomidine from the extraction procedure used in this study was greater than 75% for plasma, whole blood, or urine samples. Approximately 68% of detomidine was bound to plasma protein and 53% was bound to red blood cells.
Lehner AF, Almeida P, Jacobs J, Harkins JD, Karpiesiuk W, Woods WE, Dirikolu L, Bosken JM, Carter WG, Boyles J, Holtz C, Heller T, Nattrass C....Remifentanil (4-methoxycarbonyl-4-[(1-oxopropyl)phenylamino]-1-piperidinepropionic acid methyl ester) is a mu-opioid receptor agonist with considerable abuse potential in racing horses. The identification of its major equine urinary metabolite, 4-methoxycarbonyl-4-[(1-oxopropyl)phenylamino]-1-piperidinepropionic+ ++ acid, an ester hydrolysis product of remifentanil is reported. Administration of remifentanil HCl (5 mg, intravenous) produced clear-cut locomotor responses, establishing the clinical efficacy of this dose. ELISA analysis of postadministration urine samples readily detected fentany...
Soma LR, Uboh CE, Guan F, Moate P, Luo Y, Teleis D, Li R, Birks EK, Rudy JA, Tsang DS.The pharmacokinetics of clenbuterol (CLB) following a single intravenous (i.v.) and oral (p.o.) administration twice daily for 7 days were investigated in thoroughbred horses. The plasma concentrations of CLB following i.v. administration declined mono-exponentially with a median elimination half-life (t(1/2k)) of 9.2 h, area under the time-concentration curve (AUC) of 12.4 ng.h/mL, and a zero-time concentration of 1.04 ng/mL. Volume of distribution (V(d)) was 1616.0 mL/kg and plasma clearance (Cl) was 120.0 mL/h/kg. The terminal portion of the plasma curve following multiple p.o. administrati...
Edlund PO, Bowers L, Henion J.Monitoring steroid use requires an understanding of the metabolism in the species in question and development of sensitive methods for screening of the steroid or its metabolites in urine. Qualitative information for confirmation of methandrostenolone and identification of its metabolites was primarily obtained by coupled-column high-performance liquid chromatography-tandem mass spectrometry. The steroids and a sulphuric acid conjugate were isolated and identified by their daughter ion mass spectra in the urine of both man and the horse following administration of methandrostenolone. Spontaneo...
