• Mass Spectrometry Letters
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• v.8 no.4
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• pp.90-97
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• 2017

Androgenic anabolic steroids (AASs) are synthetic derivatives of testosterone with a common structure containing cyclopentanoperhydrophenanthrene nucleus. Their use enhances the muscle building capacity and is beneficial during performance. The AASs are one of the most abused group of substances in horse doping. Liquid chromatography tandem mass spectrometry ($LC/MS^n$) has been successfully applied to the detection of anabolic steroids in biological samples. However, the saturated hydroxysteroids viz: nandrolone, $5{\alpha}-estrane-3{\beta}$, $17{\alpha}-diol$ exhibit lower detection responses in electrospray ionisation (ESI) because of their poor ionisation efficiency. To overcome this limitation pre-column chemical derivatization has been introduced to enhance their detection responses in $LC-ESI-MS^n$ analysis. The aim of present study was to develop a sensitive method for identification and confirmation of nandrolone and its metabolite in horse urine incorporating pre-column derivatization using picolinic acid. The method consists of extraction of targeted steroid conjugates by solid phase extraction (SPE). The eluted steroid conjugates were hydrolysed by methanolysis and free steroids were recovered with liquid-liquid extraction. The resulting steroids were derivatized to form picolinoyl esters and identification was done using LC-ESI-MS/MS in positive ionization mode. The picolinated steroid adduct enhanced the detection levels in comparison to underivatized steroids.

• Journal of Animal Environmental Science
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• v.18 no.3
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• pp.201-206
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• 2012

Horse manure general composition and contaminants, heavy metals content analysis was investigated as follows. The moisture content of horse feces was 76.1% and 96% for horse urine. The average T-N, T-P, T-K concentrations of the racing horse were 0.18%, 0.10%, 0.22% respectively and 2.88%, 0.0015%, 0.84% for urine. And the average BODs, COD, SS concentrations were 26,906 $mg/{\ell}$, 36,642 $mg/{\ell}$, 89,375 $mg/{\ell}$ respectively and 14,298 $mg/{\ell}$, 7,484 $mg/{\ell}$, 6,987 $mg/{\ell}$ for urine. In addition, the Cr, Cu, Ni, Cd, Pb, Zn concentrations of feces were 2.65 mg/kg, 7.05 mg/kg, 1.24 mg/kg, 0.07 mg/kg, 0.01 mg/kg, 45.11 mg/kg respectively and 1.52 mg/kg, 1.62 mg/kg, 0.00 mg/kg, 0.03 mg/kg, 0.03 mg/kg, 4.49 mg/kg for urine on the fresh matter basis. The end of the survey was that the quality of the compost can be used as a raw material, and manure is necessary to constantly driven forward horse manure composting research.

• Journal of the Korea Organic Resources Recycling Association
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• v.16 no.2
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• pp.66-73
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• 2008

In this study, the characterization of minor livestock's excretions in terms of unit load generation and discharge was conducted by investigation and analysis of urine, manure and wastewater from stall of fowl, duck, horse, deer, goat and sheep. The results are summarized as follows: The unit load generation of fowl estimated by discharged amount and concentration analysis increases in egg layers due to the difference of planting head numbers, feed stuffs and manure disposal. In case of deer unit load generation by herbivora were calculated to be higher than data from existing references because of the gap between weight per livestock and the generation amounts of manure and urine. In case of sheep unit load generation by urine were analyzed two times higher than by manure but unit load generation by manure were reported higher than by urine in references, so large differences between this and previous study resulted.

• Analytical Science and Technology
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• v.12 no.3
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• pp.190-195
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• 1999

The metabolism of testosterone ($17{\beta}$-hydroxy-androst-4-en-3-one) was confirmed in horse after a single intramuscular administration of testosterone cypionate (750 mg). Solvent extracts of urine obtained with enzymatic hydrolysis and methanolysis were analyzed by GC/MS after oxime t-butyldimethylsilyl (oxime-TBDMS) derivatization. The structures of four urinary metabolite after testosterone administration in horse were determined based on EI mass spectra and $5{\alpha}$-androstane-$3{\beta}$, $17{\alpha}$-diol and $5{\alpha}$-androstane-$3{\beta}$-ol-17-one as major was confirmed with authentic standard. Also the concentrations of $5{\alpha}$-androstane-$3{\beta}$, $17{\alpha}$-diol, $5{\alpha}$-androstane-$3{\beta}$, $17{\beta}$-diol, dehydroepiandrosterone (DHEA), $5{\alpha}$-androstane-$3{\beta}$-ol-17-one and testosterone were determined in the urine of normal subjects and the urine after administration. The recovery and detection limit in the most drugs were 86.3~94.7% and 1~3 ppb, respectively. Correlation coefficients for calibration were in the range of 0.984~0.999. Excretion profile of testosterone presents the rapid and large increasement up to maximum values at days 5 after administration and the slow regression. The relative ratios of testosterone, its metabolites over DHEA were determined for indication of testosterone administration in horse doping.

• Biomolecules & Therapeutics
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• v.3 no.1
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• pp.80-84
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• 1995

Effects of titrated extract of Centella asiatica (TECA) and epidermal growth factor (EGF) isolated from the urine of pregnant horse on the proliferation of human epidermal keratinocyte in culture were studied. An increase in the number of keratinocyte was observed with the treatment of TECA at the concentration ranges from 1 $\mu\textrm{g}$/mι to 100 $\mu\textrm{g}$/mι. Effects of low molecular weight EGF (LEGF) and high molecular weight EGF (HEGF) on the proliferation of keratinocyte in culture were also studied. The number of keratinocyte in culture was significantly increased with LEGF and HEGF respectively at the concentration of 10 ng/mι. Simultaneous treatment of the keratinocyte with LEGF, HEGF and TECA led to the increased proliferation of keratinocytes resulting 96% of the effect of a positive control, EGF isolated from mouse submaxillary glands.

• Archives of Pharmacal Research
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• v.19 no.2
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• pp.143-147
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• 1996

LB20304 is a novel fluoroquinolone that exhibits a potent broad spectrum antibacterial activity against both gram-positve and gram-negative bacteria. The MICs (Minimal Inhibitory Concentration) of LB20304 were determined against both gram-positve and gram-negative bacteria under various conditions including several media, pHs, and inoculum concentrations. The in vitro activity of LB20304 was not significantly affected by the changes in testing conditions such as components of media and inoculum concentrations, but it was slightly reduced by acid condition. The MICs and MBCs (Minimal Bactericidal Concentration) of LB20304 against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were hardly affected by the presence of 50 % human serum, mouse serum, guinea pig serum or horse serum, and the MBCs were equal to or at most four-times higher than the MiCs. The activities of LB20304 were decreased by the presence of high concentraion of $Mg^{++}$ or human urine (pH, 5.5) in the test media. The frequencies of mutants resistant to LB20304 were similar to or lower than those found in ciprofloxacin and sparfloxacin.

• Journal of Veterinary Clinics
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• v.32 no.2
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• pp.215-217
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• 2015

There was no study on the prevalence of doping control of racehorses in Korea. The purpose of this study was to determine prohibited substances in horse races of a drugs testing program. Blood samples were taken from those 298,543 starters prior to racing and the that finished top 3 runners of each race and horses designated by the stewards shall be taken for collection of 91,482 urine samples for the purpose of post-race doping test in Seoul, Busan and Jeju Race Park between 2002 and 2013. Detection and measurement of prohibited substances were carried out by ELISA, GC/MS and LC/MS using standard methods at the Doping Control Center, Korea Racing Authority. Total 0.0030% of pre-races and total 0.0186% of post-races tested positive for prohibited substances. In pre- and post-race, caffeine and ketoprofen were the most detected prohibited substance respectively. We thought that characteristics of pharmacokinetics and pharmacodynamics of drugs resulted in different between plasma and urine. These substances have also been detected with other prohibited drugs suggesting that unintentional feeding or bedding may be the reason and groomers' confusion of candidate horses based by the stewards' research.