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.
Houghton E, Dumasia MC, Wellby JK.Negative ion chemical ionization mass spectra of some corticosteroids have been obtained by direct syringe introduction on to the Finnigan moving belt high-performance liquid chromatography-mass spectrometer interface. Proprietary preparations based upon dexamethasone, betamethasone and prednisolone were administered to horses at therapeutic dose level. Urine samples were extracted, the extracts purified by Sephadex LH-20 chromatography and the presence of the parent steroids in the eluates was confirmed by combined high-performance liquid chromatography negative ion chemical ionization mass s...
Brown MP, Stover SM, Kelly RH, Farver TB.Six healthy adult mares were given a single dose of kanamycin sulfate (200 mg/ml) IM at a dosage rate of 5 mg/kg of body weight. Kanamycin concentrations in serum, synovial fluid, peritoneal fluid, and urine were measured serially over a 48-hour period. The mean peak serum kanamycin concentration was 12.55 microgram/ml at 1 hour. Mean peak kanamycin concentrations in synovial fluid and peritoneal fluid were 7.25 microgram/ml and 9.27 microgram/ml at 2 hours and 3 hours, respectively. These concentrations decreased steadily in parallel with serum concentrations and were still measurable at 48 h...
Stover SM, Brown MP, Kelly RH, Farver TB.Six healthy adult mares were given a single dose (25 mg/kg of body weight) of sodium oxacillin IM. Oxacillin concentrations in serum, synovial fluid, peritoneal fluid, and urine were measured serially over a 48-hour period. The mean peak serum oxacillin concentration was 9.75 microgram/ml at 0.5 hour after injection. Mean peak oxacillin concentrations in synovial and peritoneal fluids were 1.45 microgram/ml and 2.60 microgram/ml at 1 hour and 2 hours, respectively. These concentrations decreased in parallel with serum values and were not measurable at 48 hours. Urine concentrations of oxacilli...
Marsh MV, Caldwell J, Smith RL, Horner MW, Houghton E, Moss MS.1. 14C-Labelled benzoic acid, salicylic acid and 2-naphthylacetic acid were administered orally to horses, and urinary metabolites investigated by chromatographic and mass spectral techniques. 2. [14C]Benzoic acid (5 mg/kg) was eliminated rapidly in the urine, and quantitatively recovered in 24 h. The major urinary metabolite was hippuric acid (95% of dose) with much smaller amounts of benzoic acid, benzoyl glucuronide and 3-hydroxy-3-phenylpropionic acid. Administration of [ring-D5]benzoic acid together with [14C]benzoic acid to a pony permitted the mass spectral determination of metabolites ...
Combie J, Blake JW, Ramey BE, Tobin T.Morphine was detected in equine biological fluids by a combination of liquid-liquid extraction and column chromatography, followed by derivatization and gas-liquid chromatographic assay, using electron capture detector. Recovery of morphine from the equine biological samples was poor. However, despite an overall recovery of less than 20%, this method had a detection limit of 0.2 ng/ml. Addition of 5,000 U of bovine liver beta-glucuronidase/ml of urine enabled detection of the drug in urine for up to 144 hours after horses were given 0.1 mg of morphine/kg of body weight. Morphine was found for ...
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...
Dumasia MC, Houghton E.1. Isomers of 3,17-dihydroxyandrostan-16-one, 3,16-dihydroxyandrostan-17-one and androstane-3,16,17-triol have been identified as urinary metabolites of testosterone in the horse. 2. Following XAD-2 extraction of urine samples, Sephadex LH-20 chromatography was used to separate the extract into conjugate groups. Metabolites obtained after hydrolysis of the conjugates have been investigated by g.l.c.-mass spectrometry. 3. Testosterone, 3,17-dihydroxyandrostan-16-one and 3,16-dihydroxyandrostan-17-one were found only in the sulphate fraction. 5 alpha-Androstane-3 beta,17 beta-diol, and two isome...
Stover SM, Brown MP, Kelly RH, Farver TB, Knight HD.Six adult mares were given a single dose of aqueous suspension procaine penicillin G (300,000 IU/ml) IM at a dosage of 22,000 IU/kg of body weight (15.4 mg of penicillin G/kg). Serum, synovial fluid, peritoneal fluid, and urine penicillin concentrations were measured serially over a 48-hour period. The mean peak serum penicillin concentration was 1.42 microgram/ml at 3 hours. Penicillin was detected in synovial fluid and peritoneal fluid, which obtained mean peak penicillin concentrations of 0.62 microgram/ml and 0.58 microgram/ml, at 4 hours and 3 hours, respectively. These concentrations ste...
Brown MP, Stover SM, Kelly RH, Farver TB, Knight HD.Six adult mares were given a single intravenous injection of oxytetracycline HCl (50 mg/ml) at a dosage of 5 mg/kg. Serum, synovial fluid, peritoneal fluid, and urine oxytetracycline concentrations were measured serially over a 48-h period. The highest measured serum oxytetracycline concentration was 8.01 mcg/ml at 1/2 h. Oxytetracycline was detected in synovial fluid and peritoneal fluid, which obtained mean peak oxytetracycline concentrations of 4.43 mcg/ml and 4.20 mcg/ml, at 1/2 h and 1 h, respectively. These concentrations steadily declined in parallel with serum concentrations and were n...
Cymbaluk NF, Schryver HF, Hintz HF.The effects of copper intake on stable copper and 64Cu metabolism and on the excretory pathways of absorbed copper were examined in ponies. Bile and feces collected from two bile duct-cannulated ponies following an intravenous dose of 64Cu showed that bile was the main route of endogenous copper excretion. Stable copper and 64Cu metabolism were examined in three intact ponies fed dietary copper to provide 5.6, 16.6 and 25.7 mg Cu/100 kg body weight/day. The amount of copper excreted in urine was not influenced by dietary treatment. Stable copper absorption and 64Cu retention significantly incr...
Cymbaluk NF, Schryver HF, Hintz HF, Smith DF, Lowe JE.The effects of molybdenum ingestion on copper metabolism were investigated in bile duct-cannulated ponies fed diets containing 1.01, 27.4 and 107.3 ppm molybdenum. Copper metabolism was assessed by examination of 64Cu and stable copper excretion in feces, bile and urine. The addition of molybdenum to the diet decreased copper absorption and retention as a consequence of increased excretion of dietary copper in feces and increased excretion of absorbed copper in bile. Only 5% or less of the absorbed copper was excreted in urine regardless of dietary treatment. Biliary molybdenum excretion was o...
Bhavnani BR, Woolever CA.[3H]Equilin [3H-labeled 3-hydroxy-1,3,5(10), 7-estratetraen-17-one] was administered iv to a pregnant mare in the 10th month of gestation. Maternal urine was collected for 3 days, and blood samples were taken 35 min and 3, 6, 12, and 24 h after the injection. The half-life of the disappearance of radioactivity from the blood was approximately 2.5 h. Over 90% of the administered dose was excreted in the first 24 h. The urine was extracted, hydrolyzed, and fractionated. The bulk of the radioactive material (75%) was present in the phenolic sulfate fraction from which radiochemically pure equilin...
Dewey EA, Maylin GA, Ebel JG, Henion JD.Promazine hydrochloride and acetylpromazine maleate were administered intravenously at clinical dose levels to horses. In urine from horses given promazine hydrochloride, the parent drug and four metabolites were detected. The two major metabolites, present as conjugates were identified after hydrolysis by beta-glucuronidase/arylsulfatase as 3-hydroxypromazine and 3-hydroxydesmonomethyl-promazine. Conjugated 3-hydroxypromazine has been previously identified as a major metabolite in the horse. Two minor metabolites isolated in this study were primaizine N-oxide and promazine N-oxide sulfoxide. ...
Frincke JM, Henderson GL.Fentanyl, a potent, synthetic narcotic analgesic, has reportedly been used to "dope" racehorses. Urine was collected from a horse dosed with 70 mg of [3H]fentanyl, and the primary metabolite, a water-soluble, amphoteric compound, was isolated by high-pressure liquid chromatography and identified by spectroscopic analysis. This metabolite was found to be N-[1-(2-phenethyl-4-piperidinyl)] malonanilinic acid.
Jonckheere JA, Thienpont LM, De Leenheer AP, De Backer P, Debackere M, Belpaire FM.A method has been developed for quantification of bromhexine in plasma using gas chromatography mass spectrometry with selected ion monitoring. A deuterium labelled analogue was synthesized and used as the internal standard. To evaluate the gas chromatographic electron capture detection method described earlier, 23 plasma samples have been analysed by both techniques. Although a good correlation was shown, selected ion monitoring was superior to the electron capture detection method for levels below 3 ng ml-1. The mass spectrometric method has also been used to set up a pharmacokinetic study o...
Chapman DI, Moss MS, Tomlinson PW, Harrison MP, Simmons PJ.1. Following single intramuscular doses of [14C]fluprostenol (0.5--2.4 micrograms/kg) to three female horses and to three gelded male horses, radioactivity was present in the plasma within 5 min; peak concn. (0.32--1.30 ng/ml fluprostenol equiv.) occurred 5 to 90 min after injection. Radioactivity was still present in the plasma of the females after three days. About 88% of fluprostenol is bound to plasma proteins. 2. Radioactivity was present in the parotid saliva of the gelded male horses within 10 min. Peak concn. (45--91 pg/ml fluprostenol equiv.) occurred from 5 min to 1 h after injection...
Henion JD, Maylin GA.A sensitive, quantitative method has been developed for the determination of hydrochlorothiazide in equine plasma and urine. Thin-layer chromatography is used to screen for the presence of the drug in unknown samples. The TLC screening methods described provide minimum detection limits of 50 ng/mL in plasma and 25 ng/mL in urine. A silica micro chromatography column is used to clean up ethyl acetate extracts for HPLC analysis and mass spectral confirmation. An internal standard, trichloromethiazide, is used to derive quantitative data at concentrations as low as 25 ng/mL for plasma disappearan...
Ray AC, Post LO, Hurst JM, Edwards WC, Reagor JC.An analytical method for cantharidin, using high performance liquid chromatography, was applied to field specimens from horses and sheep with blister beetle (Epicauta lemniscata) poisoning. Stomach content and urine proved to be valuable aids in diagnosis. One incident of cantharidin toxicosis in ruminants (sheep) was confirmed.
Houghton E, Dumasia MC.1. The metabolism of 19-nor[4-14C]testosterone in a thoroughbred horse has been studied and neutral urinary metabolites obtained after enzyme hydrolysis have been investigated by g.l.c.-mass spectrometry. 2. 3-Hydroxyestran-17-one, 17 alpha- and 17 beta-nortestosterone, estrane-3,17-diol (two isomers), 3,16-dihydroxyestran-17-one (two isomers), 3,17-dihydroxyestran-16-one (two isomers) and estrane-3,16,17-triol were identified in the neutral urinary extracts.
McKenzie RA, Blaney BJ, Gartner RJ, Dillon RD, Standfast NF.Standardbred geldings were used in a number of nutritional balance experiments. These consisted of 28-day cycles each of an 18-day equilibration period in yards and a 10-day settling and collection period in metabolism crates. The crates were made of galvanised metal tubing over wooden flooring covered by rubber matting and rubber sheets. Total faeces were collected in trays at the rear of crates and total urine by canvas funnels slung beneath the sheath of each horse.
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...
Houghton E, Dumasia MC.1. After intramuscular administration of [4-14C]testosterone to two cross-bred gelded horses, 45% of the radioactivity was excreted in urine in 96 h. Small amounts of urinary activity could still be detected at 200 h. 2. Neutral metabolites obtained after both enzyme and acid hydrolysis of urine samples have been investigated by g.l.c.-mass spectrometry. 3. 5 alpha-Androstane-3 beta, 17 alpha-diol was found only in the enzyme-hydrolysable extract and testosterone only in the acid-hydrolysable extract. 5 alpha-Androstane-3 beta, 17 beta-diol and 3 beta-hydroxy-5 alpha-androstan-17-one were foun...
Buntain B, Greig WA, Thompson H.The clinical and pathological features of a case of chronic nephritis in a 17-year-old pony was described. Measurement of fluid intake and laboratory analysis of sequential blood and urine samples helped in establishing an accurate diagnosis. The case demonstrates that although chronic renal disease is not well documented in the horse it should nevertheless be considered in the differential diagnosis of conditions characterised by progressive loss of weight.
Bhavnani BR, Woolever CA.A mixture of [4,8,12-14C]farnesyl pyrophosphate and [3H]dehydroepiandrosterone was injected into a horse fetus im during laparotomy, after which maternal urine was collected for 6 days. Steroid conjugates in the urine were extracted with Amberlite XAD-2 resin, hydrolyzed, and separated into phenolic and neutral fractions. Estrone, 17 alpha-estradiol, equilin [3-hydroxy-1,3,5(10),7-estratetraen-17-one], and 17 alpha-dihydroequilin [1,3,4(10),7-estratetraene-3,17 alpha-diol] were isolated from the phenolic fraction and their radiochemical purities were established. Only estrone and 17 alpha-estr...
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.
Blackmore DJ, Greenwood RE, Johnson C.During an investigation of a group of clinically well thoroughbreds housed in the same stable, it was noted that seven of the 14 had no detectable thyroxine in the plasma (less than 5 nmol/litre). An investigation of thoroughbreds in this stable over a five month period suggested that the thyroids were functioning normally and that the thyroxine was excreted in the urine. Thyroxine binding studies on the blood of these horses suggested that the major part of the circulating thyroxine was bound to albumin. The thyroxine had been displaced resulting in an apparent absence of plasma thyroxine sin...
de Brabander HF, van Hende J, Batjoens P, Hendriks L, Raus J, Smets F, Pottie G, van Ginkel L, Stephany RW.When residues of nortestosterone (NT) were found in the urine of cattle, racehorses or bodybuilders, exogenic administration was thought to be proven. In previous literature, no records were found of the endogenic presence of this molecule. In the horse-racing world, Houghton and Courthot found that NT is normally present in the urine of the stallion. Belgian and Dutch researchers found that NT is also present in the urine and edible parts of the intact boar. Vandenbroeck et al. (1991) suggested the endogenous presence of NT (in the beta form) in the pregnant cow. Meyer (1992) reported the pre...
Nelson EA, Sanchez LC, Mallicote MF, Warren LK, Robelen AM, Reuss SM. To compare urine urinary pH, blood pH and concentration of electrolytes in blood of healthy horses fed an anionic salt supplement to achieve diets with a dietary cation-anion difference (DCAD) of -40 or 0 mEq/kg DM, with horses a fed a diet with a DCAD of 85 mEq/kg DM. Eight healthy horses received each of three diets in a randomised crossover design. Diets consisted of grass hay and concentrate feed, with a varying amount of an anionic supplement to achieve a DCAD of 85 (control), 0 or -40 mEq/kg DM. They were fed for 14 days each with a washout period of 7 days between. Urine pH was m...
Houghton E, Teale P, Dumasia MC, Wellby JK.The negative ion chemical ionization mass spectra of the MO-TMS derivatives of the corticosteroids prednisolone, betamethasone and dexamethasone have been obtained using capillary column gas chromatography mass spectrometry. The spectra showed abundant diagnostic ions at m/z greater than 300 allowing for clear discrimination between the three steroid derivatives. A capillary column gas chromatographic mass spectrometric method using negative ion chemical ionization mass spectrometry has been developed to confirm the presence of the parent steroids in horse urine following the administration of...
Machnik M, Gerlach M, Kietzmann M, Niedorf F, Thevis M, Schenk I, Guddat S, Düe M, Schänzer W.The anti-doping rules of national and international sport federations ban any use of tetrahydrogestrinone (THG) in human as well as in horse sports. Initiated by the THG doping scandals in human sports a method for the detection of 3-keto-4,9,11-triene steroids in horse blood and urine was developed. The method comprises the isolation of the analytes by a combination of solid phase and liquid-liquid extraction after hydrolysis and solvolysis of the steroid conjugates. The concentrations of THG in blood and urine samples were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS)...
Kusano K, Minamijima Y, Mashita S, Kunii H, Yamashita S, Nagata S.Repeated topical application of indomethacin is common in Japanese racehorses, despite the lack of pharmacokinetic data. Objective: To determine the concentrations of indomethacin and its metabolite, desmethylindomethacin, in plasma and urine of Thoroughbreds topically treated repeatedly with indomethacin. Methods: In vivo experimental. Methods: Seven female Thoroughbreds were topically treated with 50 g of 1% indomethacin cream per horse to the back and hips (500 mg of indomethacin/head/2400 cm , 0.21 g/cm ) for 3 consecutive days. Samples were pretreated by protein precipitation for plas...
Dias DCR, Ribeiro Filho JD, Viana RB, Bittencourt TCC, Dantas FTDR, Teixeira RBC, Di Filippo PA, Manso Filho HC, Alves SR, Santos PVM, Moreira NS.Continuous flow enteral fluid therapy with isotonic and hypotonic enteral electrolyte solutions are as safe and effective as intravenous fluid therapy. The aim of this study was to carry out a comparative assessment between continuous flow enteral and intravenous (IV) fluid therapy in adult experimentally dehydrated horses. Six experimentally dehydrated adult mares were used in a study carried out in a 6 × 3 crossover design, which each animal received three different treatments (isotonic enteral fluid therapy-EsISO, hypotonic enteral fluid therapy-EsHYPO and intravenous fluid therapy with La...
Richards SL, Cawley AT, Raftery MJ.Dermorphin, a hepta-peptide with potent analgesic properties, is classified as a doping agent in equine racing. Since its discovery, a number of biologically active structural analogs have been synthesized and made commercially available so there is a need for reliable methods of detection. Results: A sensitive detection method was developed for dermorphin and six analogs in equine urine. Peptide enrichment was achieved using weak cation exchange with subsequent separation and detection by nano-UHPLC-MS/MS. Method validation parameters included: specificity, linearity (5-10000 pg/ml), recovery...
Vander Werf KA, Davis EG, Kukanich B.The objective of this study was to assess the pharmacokinetic profile and determine whether any adverse effects would occur in seven healthy adult horses following oral meloxicam tablet administration once daily for 14 days at a dose of 0.6 mg/kg·bwt. Horses were evaluated for health using physical examination, complete blood count, serum chemistry, urinalysis, and gastroscopy at the beginning and end of the study. Blood was collected for the quantification of meloxicam concentrations with liquid chromatography and mass spectrometry. The mean terminal half-life was 4.99 ± 1.11 h. There was n...
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...
Arosalo BM, Raekallio M, Rajamäki M, Holopainen E, Kastevaara T, Salonen H, Sankari S.The aim of the study was to investigate urine matrix metalloproteinase (MMP-2 and -9) activity, alkaline phosphatase/creatinine (U-AP/Cr) and gamma-glutamyl-transpeptidase/creatinine (U-GGT/Cr) ratios, glucose concentration, and urine protein/creatinine (U-Prot/Cr) ratio and to compare data with plasma MMP-2 and -9 activity, cystatin-C and creatinine concentrations in colic horses and healthy controls. Horses with surgical colic (n = 5) were compared to healthy stallions (n = 7) that came for castration. Blood and urine samples were collected. MMP gelatinolytic activity was measured by zymogra...
Soma LR, Behrend E, Rudy J, Sweeney RW.The disposition of flunixin meglumine administered IV at a dosage of 1.1 mg/kg was described by a 2-compartment model; the alpha and beta half-lives (t1/2) were 0.61 and 1.5 hours, respectively. When administered IV at a rate of 2.2 mg/kg, the disposition was best described by a 3-compartment model, and the alpha, beta, and lambda t1/2 were 0.16, 1.52, and 6.00 hours, respectively. The zero-time plasma concentrations after flunixin meglumine was administered at 1.1 and 2.2 mg/kg were 9.3 +/- 0.76 and 21.5 +/- 7.4 mg/L, respectively. The bioavailability after oral administration of 1.1 mg/kg wa...
Vonaparti A, Lyris E, Panderi I, Koupparis M, Georgakopoulos C.In equine sport, theobromine is prohibited with a threshold level of 2 microg mL(-1) in urine, hence doping control laboratories have to establish quantitative and qualitative methods for its determination. Two simple liquid chromatography/mass spectrometry (LC/MS) methods for the identification and quantification of theobromine were developed and validated using the same sample preparation procedure but different mass spectrometric systems: ion trap mass spectrometry (ITMS) and time-of-flight mass spectrometry (TOFMS). Particle-free diluted urine samples were directly injected into the LC/MS ...
You Y, Uboh CE, Soma LR, Guan F, Li X, Rudy JA, Chen J.A rapid and sensitive method was developed for the screening, quantification and confirmation of ethyl glucuronide (EG) and ethyl sulfate (ES) as biomarkers for alcohol administration to racehorses using liquid chromatography coupled on-line with triple quadrupole tandem mass spectrometry. Urine sample aliquots (0.1 mL) were pre-treated by protein precipitation. Separation of EG and ES was achieved on an Ultra PFP column. Isocratic elution with a flush step was performed using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). Analysis was performed by negative electrospra...
Schenk I, Broussou D, Roques B, Lagershausen H, Machnik M, Röttgen H, Toutain PL, Thevis M.The combination of sulfadoxine (SDO) with trimethoprim (TMP) is widely used in veterinarian medicine. The aim of the present study was to compare excretion profiles and detection time windows of SDO and TMP in plasma and urine by means of a validated quantitative method. Eight horses received a single intravenous (i.v.) dose of 2.7 mg TMP and 13.4 mg SDO per kg bodyweight. Plasma and urine samples were collected up to 15 and 70 days post-administration, respectively. While urine samples underwent an enzymatic hydrolysis, plasma samples were proteolysed before further analysis. After solid-...
Carstanjen B.Effective, non-invasive bone assessment methods for screening, diagnosis and follow-up of the skeleton are more and more requested in veterinary medicine. In contrast to clinical parameters, invasive methods and imaging techniques, indices of bone turnover is a tool for bone metabolism evaluation of the whole skeleton. Biochemical bone markers therefore provide a more real-time assessment of the bone status with simple blood- or urine-analysis. This article surveys currently available biochemical marker of bone metabolism used in veterinary medicine. Additionally, information is provided about...
Wenzel RG, Major D, Hesp KF, Hall E, Doble P.To monitor cobalt concentrations in urine, red blood cells and plasma after chronic parenteral administration of cobalt chloride evaluate these results against the current International Federation of Horseracing Authorities thresholds for detecting cobalt misuse. Methods: Eight mares were randomly assigned to four treatment groups, with two mares in each group: Group 1 - control group, Group 2 - 25 milligrams cobalt intravenously as CoCl weekly, Group 3 - 50 milligrams cobalt intravenously as CoCl weekly, and Group 4 - 25 milligrams cobalt intravenously mid-week and at the end of the week. Uri...
Colahan PT, Bailey JE, Chou CC, Johnson M, Rice BL, Jones GL, Cheeks JP.Twelve clinically sound, healthy, athletically conditioned Thoroughbred horses were subjected to an incremental exercise stress test to determine the effects and period of detection of a single dose of flunixin meglumine (1.1 mg/kg by intravenous injection) in serum and urine by ELISA. Flunixin concentrations, performance, and hematologic and clinical chemical parameters were measured. All horses were rotated through four treatment groups of a Latin-square design providing for each horse to serve as its own control. Flunixin meglumine reduced prostaglandin F(1alpha) and thromboxane concentrati...
Ho EN, Kwok WH, Wong AS, Wan TS.Quaternary ammonium drugs (QADs) are anticholinergic agents some of which are known to have been abused or misused in equine sports. A recent review of literature shows that the screening methods reported thus far for QADs mainly cover singly-charged QADs. Doubly-charged QADs are extremely polar substances which are difficult to be extracted and poorly retained on reversed-phase columns. It would be ideal if a comprehensive method can be developed which can detect both singly- and doubly-charged QADs. This paper describes an efficient liquid chromatography/tandem mass spectrometry (LC/MS/MS) m...
Hagedorn HW, Schulz R, Jaeschke G.An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone-17-hemisuccinate-bovine serum albumin as antigen. Boldenone-17-hemisuccinate-horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 +/- 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross-reactivities, 3.4 and 2.5%, respectively. These cross-reactivities are of no importance for the boldenone assay. For the reductio...
Henion JD, Maylin GA.A sensitive, quantitative method has been developed for the determination of hydrochlorothiazide in equine plasma and urine. Thin-layer chromatography is used to screen for the presence of the drug in unknown samples. The TLC screening methods described provide minimum detection limits of 50 ng/mL in plasma and 25 ng/mL in urine. A silica micro chromatography column is used to clean up ethyl acetate extracts for HPLC analysis and mass spectral confirmation. An internal standard, trichloromethiazide, is used to derive quantitative data at concentrations as low as 25 ng/mL for plasma disappearan...
Divers TJ, Chang YF, Irby NL, Smith JL, Carter CN.North American horses are commonly exposed to Leptospira organisms. Leptospira Bratislava is the most common infecting serovar but this serovar has not been confirmed to cause clinical disease in North American horses. Leptospira Pomona type kennewicki is responsible for most of the clinical diseases (leptospirosis) in North American horses. Leptospirosis is most commonly associated with diseases of the placenta and fetus, the kidneys and the eyes in horses. In-utero infections in pregnant mares may result in abortion, neonatal illness or birth of an antibody positive healthy foal. Acute renal...
Garcia P, Paris AC, Leufroy A, Popot MA, Bonnaire Y.A quantitative method, using LC/ESI-MS(n) with a quadrupole linear ion trap mass analyzer, has been developed for the analysis of ipratropium cation in horse plasma and urine. The method applies solid-phase extraction with WCX cartridges for plasma and MM2 cartridges for urine, prior to analysis by LC/ESI-MS(n). The efficiency of extraction combined with the sensitivity and the selectivity of MS(n) allows for the quantification of ipratropium cation at picogram per milliliter levels. The analytical capabilities of the method have been successfully checked by the quantitative analysis of ipratr...
Elfenbein JR, Giguère S, Meyer SK, Javsicas LH, Farina LL, Zimmel DN, Sanchez LC.Hepatic failure is one of the more common complications in foals requiring blood transfusion to treat neonatal isoerythrolysis. Iron intoxication is likely the cause of hepatic injury. Objective: To determine the effects of deferoxamine on iron elimination in normal foals. Methods: Thirteen neonatal foals. Methods: Randomized-controlled trial. At 1-3 days of age, foals received either 3 L of washed packed dam's red blood cells (RBC) or 3 L of saline IV once. Foals were treated with deferoxamine (1 g) or saline (5 mL) SC twice daily for 14 days. Foals were randomly assigned to 1 of 3 gr...
Dirikolu L, Lehner AF, Hughes C, Karpiesiuk W, Camargo FC, Harkins JD, Woods WE, Bosken JM, Boyles J, Troppmann A, Fisher M, Tobin T.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 (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 th...
Hagedorn HW, Meiser H, Zankl H, Schulz R.The misuse of opiates in racehorses relates to their effect of increasing locomotor activity. Because methadone, a narcotic analgesic, has been suspected of use as a doping compound in the past, it was added to the list of banned drugs and should be considered in doping control. Because the literature fails to provide information on detection of methadone in blood or urine of horses, an enzyme-linked immunosorbent assay was developed to monitor this narcotic in equine body fluids. Combined with high-performance liquid chromatography, the immunoassay also served to confirm positives indicated b...
Lavy E, Ziv G, Glickman A.An aqueous solution of norfloxacin nicotinate (NFN) was administered to donkeys (Aquus asinus) intravenously (once at 10 mg/kg), intramuscularly and orally (both routes once at 10 and 20 mg/kg, and for 5 days at 20 mg/kg/day). Blood samples were collected at predetermined times after each treatment and urine was sampled after intravenous drug administration. Serum NFN concentrations were determined by microbiological assay. Intravenous injection of NFN over 45-60 s resulted in seizures, profuse sweating and tachycardia. The intravenous half-life (t1/2 beta) was 209 +/- 36 min, the apparent vol...
Ferrante PL, Freeman DE, Ramberg CF, Kronfeld DS.Multicompartmental analysis was applied to study the kinetics of D-xylose distribution after IV administration to healthy mares deprived of food for 12 and 96 hours. Urinary excretion of D-xylose was measured over a 15-hour period after administration. The plasma D-xylose concentrations in this study were in the range found after oral tolerance testing. The disposition of D-xylose was described by a two-compartment model with linear kinetic characteristics. Total volume of distribution decreased significantly (P < 0.025) from 0.270 L/kg of body weight after the 12-hour period of food depriv...
Chuang MS, Huang HH, Dixon KM, Chen KS, Mao CL, Chen CL.The pharmacokinetics of clenbuterol in equine urine and blood was investigated. Methods: Urine and blood samples were collected following 3-day multiple oral administrations. The samples were examined using enzyme-linked immunosorbent assay and further confirmed by solid phase extraction and capillary electrophoresis. Results: Urinary clenbuterol was detectable until day 14 after the last dose. The urinary excretion of clenbuterol was characterized by a biphasic pattern. The half-lives of the bi-exponential elimination (t(1/2alpha) and t(1/2beta)) for urinary clenbuterol were about 12.1 and 48...
Stanley S, Jeganathan A, Wood T, Henry P, Turner S, Woods WE, Green M, Tai HH, Watt D, Blake J.We have raised antibodies to morphine and etorphine and developed one-step enzyme-linked immunosorbent assays (ELISA) for these drugs as part of a panel of post race tests for drugs in racing horses. These tests are simple, can be completed in 2 h, and can be read by visual inspection. The morphine ELISA has an I50 for morphine of about 1.5 ng/mL, while the etorphine ELISA has an I50 for etorphine of 250 pg/mL. Cross-reactivity studies show that the antimorphine antibody cross-reacts well with levorphanol, hydromorphone, and oxycodone, while the anti-etorphine antibody showed no cross-reactivi...
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...
