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Topic:Analytical Methods
Analytical methods in equine research encompass a variety of scientific techniques and tools used to study and evaluate different aspects of horse health, performance, and physiology. These methods help advance our understanding of equine biology, diagnosing conditions, and improving management practices. Common analytical methods include molecular techniques like PCR and ELISA for detecting pathogens and measuring biomarkers, imaging technologies such as ultrasound and MRI for assessing musculoskeletal health, and statistical models for analyzing genetic data and performance metrics. This page compiles peer-reviewed research studies and scholarly articles that explore the development, application, and impact of various analytical methods in equine science.
Pharmacology of narcotic analgesics in the horse: quantitative detection of morphine in equine blood and urine and logit-Log transformations of this data. 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 ...
Effects of dilution rates, animal species and instruments on the spectrophotometric determination of sperm counts. Using semen from bull, boar and stallion as well as different spectrophotometers, we established the calibration curves relating the optical density of a sperm sample to the sperm count obtained on the hemacytometer. The results show that, for a given spectrophotometer, the calibration curve is not characteristic of the animal species we studied. The differences in size of the spermatozoa are probably too small to account for the anticipated specificity of the calibration curve. Furthermore, the fact that different dilution rates must be used, because of the vastly different concentrations of ...
Studies related to the metabolism of anabolic steroids in the horse: the identification of some 16-oxygenated metabolites of testosterone and a study of the phase II metabolism. 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...
Helix packing and subunit conformation in horse spleen apoferritin. An electron density map of horse spleen apoferritin at 0.28-nm (2.8 A) resolution and its preliminary interpretation have been described previously. Rigorous examination of this and newer maps at the same nominal resolution but calculated from more extensive data sets, including model building in a Richards' comparator, now allows us to report on structural features in more detail. We list inter-helical angles within and between neighbouring subunits, and describe a new short region of inter-subunit anti-parallel pleated sheet. A short section of electron density not properly accounted for in ...
Selected ion monitoring assay for bromhexine in biological fluids. 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...
Characterization of gangliosides from equine kidney and spleen. Gangliosides were isolated from equine kidney and spleen, and their carbohydrate and lipid moieties were characterized. Among the long-chain bases, considerable proportions of trihydroxy bases (42.3 to 61.2% of the total bases), in which phytosphingosine was predominant were found in all the ganglioside classes. The other major base was sphingosine. Among the constituent fatty acids, long-chain acids (with a carbon number of more than 20), comprised approximately half the total acids, with some alpha-hydroxy and mono-unsaturated acids. By means of sequential hydrolysis with glycosidases couple...
Horse erythrocyte gangliosides: preparation of the major hematoside NeuNG1-Lac-Cer. A simple method for the isolation of hematoside NeuNG1-Lac-Cer from horse erythrocytes is described. An aliquot of the crude ganglioside fraction was labeled by tritiated sodium borohydride after mild periodate oxidation. The compounds obtained were used as radioactive tracers in column chromatography. Gangliosides were applied onto a silicic acid column and eluted stepwise by solvents of steadily increasing polarity. The major ganglioside, NeuNG1-Lac-Cer, was eluted in a high yield by the solvent mixture chloroform/methanol/water (60:35:8, v/v/v).
Qualitative and quantitative analysis of hydrochlorothiazide in equine plasma and urine by high-performance liquid chromatography. 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...
Evaluation of an analytical method for the diagnosis of cantharidin toxicosis due to ingestion of blister beetles (Epicauta lemniscata) by horses and sheep. 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.
Skin surface lipids of the horse. Skin surface lipids from the sides of male and female horses (Equus caballus) were collected in acetone and analyzed by thin layer chromatography and gas liquid chromatography. The sole components in both sexes were cholesterol, cholesteryl esters and the lactones of 32-, 32- and 36-carbon omega-hydroxy acids, each including a methyl group in the n-1 position. Most of the lactones were monounsaturated (either n-8 or n-10), but small amounts of saturated and dienoic species were present. A pooled sample of the skin surface lipids contained 14% cholesterol, 38% cholesteryl esters and 48% lactone...
Serum quality: an analysis of its components. Foetal and new born bovine sera, horse serum, human serum and human plasma, and protein solutions prepared from the by-products of human plasma fractionation have been analysed. Foetal bovine sera were found to have lower total protein (g/l) and % of gamma-globulin than the other sera studied while the potassium (mmol/1) was higher. Protease inhibitors could be detected in all specimens tested.
A gas chromatographic screening procedure for the detection of non-steroidal anti-inflammatory drugs in horse urine. 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...
The effect of binding ions on the oxidation of horse heart ferrocytochrome c. The research explores how different binding ions affect the oxidation speed of horse heart ferrocytochrome c, a protein, by potassium ferricyanide at a constant ionic strength. Studying the Ion Effect […]
Molecular cytogenetics of the Equidae. I. Purification and cytological localization of a (G + C)-rich satellite DNA from Equus przewalskii. A (G + C)-rich density satellite DNA (rho = 1.713 gm/cc) has been purified from splenic DNA of Przewalski's horse, Equus przewalskii, by successive equilibrium density gradient centrifugations. The purified satellite, which may comprise as much as 29% of the total DNA, renatures rapidly; however, analyses of native, single-stranded, and reassociated molecules by analytical ultracentrifugation and melting properties suggest that some sequence heterogeniety exists in the 1.713 gm/cc satellite. Complementary RNA (cRNA) transcribed from satellite DNA has been utilized for in situ hybridization stu...
High pressure liquid chromatographic determination of cantharidin, using a derivatization method in specimens from animals acutely poisoned by ingestion of blister beetles, Epicauta lemniscata. Experimental animals (rabbit, rat, goat, sheep, and pony) were given cantharidin or dried preparations of blister beetles (Epicauta lemniscata) to stimulate naturally occurring toxicosis in which beetles were ingested with alfalfa hay. A sensitive high-pressure liquid chromatographic method, involving derivatization of cantharidin with p-nitrobenzyloxyamine, was developed to detect the toxin extracts of ingesta, fluids, and tissues from these severely poisoned animals. Urine and ingesta from the upper portion of the gastrointestinal tract, containing from 1 to 20 ppm of cantharidin, were the m...
Insensitivity of the ferritin iron core to heat treatment. To test whether the reactivity of ferritin iron is affected by the heat treatment used in ferritin isolation, we prepared ferritin from the same horse spleen with or without heating. Both samples exhibited similar reactivity upon reduction or chelation of iron.
Determination of the charge of horse kidney metallothionein by free boundary electrophoresis. Traditionally, the charge of a protein molecule as determined by electrophoresis has been compared to that revealed by pH titration, and any lack of coincidence has been ascribed to ion binding, and the two results have been brought into agreement by adjustment of binding parameters (1). Metallo-thionein allows a unique opportunity to examine the validity of the electrophoretic approach, since the amino acid sequence and metal atom binding studies allow the absolute charge of the molecule to be computed (2). This then can be compared to the charge determined from electrophoretic mobility measu...
A detection tube for cholinesterase inhibiting compounds. The enzyme butyrylcholinesterase from horse serum catalyses the hydrolysis of certain esters. The orange-red 2,6-dichloroindophenyl acetate will be converted by the enzyme into a deep blue alcohol. The colour transformation does not occur when the enzyme is inactivated. By making use of this biochemical reaction a cheap and simple, but very sensitive and specific detection tube could bedeveloped. The tube comprises a breakable ampoule with an aqueous buffer solution, a freeze-dried preparation of the chromogenic ester with a filler promoting its dissolution, a freeze-dried preparation of butyr...
Radioimmunoassay of oxfendazole in bovine, equine, or canine plasma or serum. A simple radioimmunoassay was developed for the determination of oxfendazole in plasma. Oxfendazole N-1(3)-valerate was coupled to polylysine via a carbodiimide reaction, and antiserum was developed in rabbits after inoculation with oxfendazole--polylysine conjugate. The assay was developed so that oxfendazole could be measured directly in a 0.1-ml aliquot of diluted or undiluted plasma. With the developed procedure, 200 pg of oxfendazole/ml of plasma can be determined quantitatively. Cross-reactivity was determined for closely related compounds and metabolites. The method was used to determin...
The presence of two (Na+ + K+)-ATPase inhibitors in equine muscle ATP: vanadate nad a dithioerythritol-dependent inhibitor. A potent inhibitor of (Na+ + K+)-ATPase activity was purified from Sigma equine muscle ATP by cation- and anion-exchange chromatography. The isolated inhibitor was identified by atomic absorption spectroscopy and proton resonance spectroscopy to be an inorganic vanadate. The isolated vanadate and a solution of V2O5 inhibit sarcolemma (Na+ + K+)-ATPase with an I50 of 1 micrometer in the presence of 1 mM ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA), 145 mM NaCl, 6mM MgCl2, 15 mM KCl and 2 mM synthetic ATP. The potency of the isolated vanadate is increased by free Mg2+. ...
Clinical signs and chemical confirmation of 4-aminopyridine poisoning in horses. 4-Aminopyridine poisoning in horses was diagnosed. Specific methods, utilizing thin-layer and high-performance liquid chromatography, were developed for determining the compound in stomach contents and corn bait. The lethal dose was estimated at 2 to 3 mg/kg of body weight.
