• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.17-29
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• 2004

A multiresidual analysis was performed to determine 12 sulfonamides(sulfacetamide, sulfadiazine, sulfisomidine, sulfathiazole, sulfapyridine, sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfisoxazole, sulfamethoxazole, sulfaquinoxaline, and sulfadimethoxine) in beef and pork simultaneously. The multiresidual analysis for the sulfonamides currently used was able to analyze 5 kinds of sulfonamides at the same time. The method of this 12 sulfonamides multiresidual analysis in this study was matrix solid-phase dispersion(MSPD) by high performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS). The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM: Ethylacetate(3:1), DCM:EA(9:1). Also, samples (84 beef and 205 pork samples) which were positive by EEC-4 plate test from 2001 to 2003 were tested to investigate the kinds of sulfonamides using HPLC. The results from the study were as follows; 1. The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM:Ethylacetate(3:1), DCM:EA(9:1). The method of extraction solvent with DCM:ethyl acetate(9:1) was the most excellent(87.7∼99.3%) in separation and reappearance. 2. In the LC/MS analysis. of sulfonamides, signal to noise ratio was showed relatively high in the positive mode and special ion in the quality analysis was determined via [M+H]$\^$+/ and m/z 156. A spectrum of sulfonamides was showed from all 12 sulfonamides. 3. The samples positive by the EEC-4 plate, a screening test method, were categorized by sulfonamides through Charm II and confirmed the kinds of sulfonamides through HPLC. 1) Among 84 beef samples positive by EEC-4 plate, 20 samples were positive by Charm II and identified as 7 sulfamethazine, 9 sulfadimethoxine, 1 sulfamonomethoxine and 3 unknown status. 2) Among 205 pork samples positive by EEC-4 plate, 42 samples were positive by Charm II and identified as 19 sulfamethazine, 1 sulfadimethoxine, 4 sulfamonomethoxine and 5 unknown status.

• Korean Journal of Veterinary Service
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• v.20 no.4
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• pp.339-348
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• 1997

The purpose of the survey were determined the residual antibiotics and tetracyclines in beef(n=1,364), pork(n=2,817) and chickens(n=1,921) by the EEC 4-plate method, Charm IIand HPLC. The results were summarized as follows ; 1. The recovery rates in spiked samples were 98.8-107.2% for oxytetracycline(OTC), 33.2-48.6% for tetracycline(TC) and 64.1-72.3% for chlortetracycline(CTC) at 0.05-0.1ppm by HPLC using MSPD. 2. Residues of antibiotic were detected from 10 beef(0.75%) and 36 pork(1.31%) by EEC 4-plate method. In case of chickens were not detected. 3. Twenty-eight from 46 positive samples by the EEC 4-plate method were classified as TCs(60.9 %) by Charm II and TC of 12 samples were detected by HPLC. 4. Ten samples were detected levels of OTC ranging from 0.035 to 0.635 ppm and 3 samples were levels of CTC ranging from 0.066 to 0.150 ppm. OTC and CTC levels in 3 beef and 4 pork samples were exceeded the current tolerance level of 0.1 ppm.

• Korean Journal of Veterinary Service
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• v.27 no.3
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• pp.265-272
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• 2004

It was carried out to compare the residual materials by EEC 4-plate, Charm II and HPLC method in the muscles of cattle and pigs from slaughter-houses in Seoul from 2000 to 2003. Residual materials were detected from $1.10\%$(73/6,623) samples by EEC 4-plate method, and $10.93\%$(55/503) samples by Charm II method. The highest residual concentration(ppm) of oxytetracycline, tetracycline, chloramphenicol, chlortetracycline, sulfamethazine, sulfamerazine, sulfadimethoxine, penicillin and sulfamonomethoxine were 25.5, 3.46, 3.26, 1.5, 0.3, 0.2, 0.2, 0.14, and 0.07, respectively. Eighty nine samples were classified as 58($65.17\%$) only tetracyclines, 20($22.47\%$) only sulfonamides, 3($3.37\%$) only ${\beta}$-lactams, 2($2.25\%$) only chloramphenicol, 4($4.49\%$) tetracyclines and sulfonamides simultaneously, 1($1.12\%$) chloramphenicol and sulfonamides simultaneously, and 1($1.12\%$) chloramphenicol, sulfonamides and tetracyclines simultaneously.

• Korean Journal of Veterinary Service
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• v.25 no.3
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• pp.285-294
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• 2002

The present studies were carried out to determine antibiotics residues in pork and beef muscles by EEC-4-plate and HPLC. A total of 2,534 samples of pork muscles and 1,070 samples of beef muscles from slaughter houses were collected in Gyeongnam area from January to December, 2001. The results were summarized as follows; 1. Recovery rates of TCs, Sulfa drug, Penicillin G from fortified pork and beef muscles ranged as 68.79～98.24%, 78.21～94.58% and penicillin G 63.35～67.24% respectively, by HPLC. 2. Antibiotics residues were detected in 36 sample(1.42%) of pork muscles, 29 sample (2.71%) of beef muscles by EEC-4-plate. 3. Detection rate of antibiotic residues 14 samples(0.55%) and 26 samples(2.43%), in pork and beef muscles, respectively by HPLC. Concentration of residues in 22 sample(2.06%) of beef muscle were higher than tolerance level in korea. 4. Antibiotics detected were sulfamethazine(47.37%), tetracycline(15.79%), oxytetracycline (15.79%), penicillin G(15.79%), sulfamerazine(5.26%) in pork muscle samples and oxyteracycline (37.21%), penicillin G(30.23%), sulfamethazine(20.93%), tetracycline(4.65%), sulfamerazine (2.33%), sulfadimethoxine(2.33%), sulfaquinoxine(2.33%) in beef muscle samples.

• Korean Journal of Veterinary Service
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• v.26 no.2
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• pp.113-119
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• 2003

This study was carried out to compare the antibiotic residues in the muscles of cattle and pigs from slaughter-houses in Seoul from 2000 to 2002 by EEC-4-plate, Charm II and HPLC methods. The results were summarized as follows; 1. Residual materials were detected from 41 samples(0.6%) by EEC-4-plate method from random sampling and 38 samples(12%) by Charm II method from directed sampling. 2. Violation rates were 0.3% by monitoring and 4.7% by surveillance program. 3. The 35 samples were classified as tetracyclines 30(86%), sulfonamides 4(11%), ${\beta}$-lactams 1(3%) and two samples simultaneously determined oxyteracycline plus sulfadimetoxine, and sulfamerazine plus sulfadimetoxine. 4. The highest residual concentration(ppm) of chlortetracycline, oxytetracyline, sulfamethazine, sufadimetoxine and penicillin were 0.5, 12.0, 6.4, 2.6 and 0.44, respectively.

• Korean Journal of Veterinary Service
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• v.15 no.2
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• pp.93-100
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• 1992

The purpose of the present survey was to evaluate the antibiotic residues in meats and internal organs such as muscle, liver, heart, kidney and spleen of cattle (n=59) and pigs (n=115). The EEC-4-plate-method were employed. The results were obtained as follows ; 1. In BS 6.0, BS 7.2 and BS 8.0 used as media to detect antibiotic residues, the zone($M{\pm}SD,$ cm) of bacterial growth inhibition was narrow($1.40{\pm}0$) in meats, whereas the zone was wide($1.69{\pm}0.25-1.88{\pm}0.23$ and $1.58{\pm}0.18-1.86{\pm}0.15$ in cattle and pigs, respectively) in internal organs. But in SL 8.0, it was difficult to detect the zones ($0-1.40{\pm}0$) of both meats and internal organs. 2. Residues of antibiotic in beef and pork were rarely detected in BS 6.0, BS 7.2 and BS 8.0 (range 1.7-11.9% and 2.6-4.3%, respectively), whereas residual percentages of internal organs were relatively higher(range 69.5-96.6% and 43.5-84.3%, respectively). But in SL 8.0, it was not detected in both beef or pork, whereas they were 0-13.6% and 0-4.3% in interanal organs.

• Journal of fish pathology
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• v.11 no.1
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• pp.77-81
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• 1998

By standardized method, Bacillus subtilis BGA, Bacillus cereus var. mycoides ATCC 11778, and Micrococcus luteus ATCC 9341 were seeded on the muller hinton agar (Difco) plate, and pH was adjusted to 6.0, 7.2, and 8.0. Five agar plates, B. subtilis (pH 6.0), B. cereus (pH 6.0), B. subtilis (pH 7.2), B. subtilis (pH 8.0), and M. luteus (pH 8.0), were employed as test plates of modified EEC 4-plate method. Oxytetracycline (OTC) with a diet was orally administered to flounder, Paralichthys olivaceus, at 100 mg/kg once a day. After oral administration, modified EEC 4-plate method by the three screening test using muscle-direct, extraction-disk and direct-disk methods was conducted for 3 fish at 1, 3, 5, 10, 15, 20, 25 and 30 days. Muscle-direct treatment of B. subtilis (pH 6.0) was found to be dubious positive (${\pm}$) at the 1st day after the administration; thereafter, it was found to be negative to the last day of the experiment. Extraction-disk and direct-disk treatment of B. subtilis (pH 6.0) were found to be negative from the 1st day to the last day after the administration. B. subtilis (pH 7.2), B. subtilis (pH 8.0), and M. luteus (pH 8.0) by the three screening tests, were found to be negative all the way after the administration. On the other hand, B. cereus (pH 6.0) by the three screening tests was clearly found to be positive for the first 15 days after the administration, and then muscle-direct and direct-disk treatment of B. cereus (pH 6.0) were found to be dubious positive at 20th days after the administration. However extraction-disk treatment of B. cereus (pH 6.0) was clearly found to be negative at the same stage; thereafter, the three screening tests of B. cereus (pH 6.0) were found negative to the last of the experiment. These findings showed that to have equal sensitivity to those determination for the residual detection of OTC, and also confirmed that B. cereus was effective test organism for the monitoring of OTC.

• Journal of Food Hygiene and Safety
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• v.19 no.1
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• pp.12-18
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• 2004

Ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin in chicken muscle were seperated by liquid extraction and determined with high performance liquid chromatography (HPLC) with fluorescence detector. Analysis was carried out using following conditions; Cl8 column (250${\times}$4.6 mm i.d. 5 ${\mu}{\textrm}{m}$ particle size), mobile phase composed of D.W. (containing 0.4% triethylamine and phospholic acid): methanol : acetonitrile (800:100:100, v/v/v), isocratic pump at a flow rate of 1.0 $m\ell$/min and 50 ${mu}ell$ of injection volume, fluorescence detector with EX278 nm/EM.456 nm. The calibration curves of four fluoroquinolones showed linearity (${\gamma}$$^2$$\geq$0.999) at concenration range of 0.025-0.6 $\mu\textrm{g}$/ml. The recoveries in fortified chicken muscle represented more than 80% with low coefficient of variation (〈10%) for concentration range of four fluoroquinolones. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin were 23.5, 3.4, 3.0 and 2.5 ng/g in chicken muscle, respectively. We also monitored fluoroquinolones residue in muscle of chickens (broiler 1:227, Korean native chicken 219, laying chicken 77) using EEC-4-plate screening and HPLC conformation methods. Ten(broiler 5, Korean native chicken 5) out of the fifteen samples which were positively detected by EEC-plate screening method from 1,523 chicken meat were confirmed with ciprofloxacin and enrofloxacin by HPLC. The ranges of residual concentration were 0-0.12 ppm for ciprofloxacin and 0.01-6.79 ppm for enrofloxacin. In conclusion, our method could be applied effectively to determine four fluoroquinolones residues in chicken meat, and further survey for fluoroquinolones residue in chicken meat are needed for more effective control of fluoroquinolones used in livestock.

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.281-286
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• 2009

Fluoroquinolones in muscle and egg were separated by liquid extraction and determined. The analysis was carried out using following conditions; C18 column ($150{\times}4.6mm$, $5{\mu}m$), mobile phase composed of D.W. (containing 0.4% triethylamine and phosphoric acid) : methanol : acetonitrile (780:100:120, v/v/v), quarternary pump at a flow rate of 0.9ml/min and $20{\mu}l$ of injection volume, fluorescence detector with EX 278nm/Em 456nm. The calibration range of seven fluoroquinolones showed linearity ($r^2{\geq}0.999$) at concentration range of $0.025{\sim}0.8{\mu}g/ml$. The recoveries in fortified muscle and egg represented more than 81.3%. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin, saraloxacin and orbifloxacin were 3.1, 2.5, 3.6, 1.7, 0.9, 2.5 and $2.1{\mu}g/kg$, respectively. We also monitored fluoroquinolones residue in the sample (chicken muscle 182, cattle muscle 140, pig muscle 139, egg 212) using EEC-plate (E. coli ATCC 11303) screening and HPLC confirmation methods. The screening test results, fluoroquinolones, antibacterial substances were all negative.