• Journal of Food Hygiene and Safety
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• v.19 no.4
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• pp.198-202
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• 2004

It was carried out to compare the relationship of EEC 4-plate test method and Charm II test method by HPLC. 46 only tetracyclines, 4 only sulfonamides, 4 sulfonamides+tetracyclines, one amphenicols+tetracyclines, and one amphenicols+sulfonarnides+tetracyclines, it was detected on 71 porks of EEC 4-plate positive or Charm II positive by HPLC. In case of sulfonamides and chloramphenicols, two test methods had related with each other, but had no relation in tetracyclines. Also, it was $10.5\%(6/57)$ tetracyclines, $81.8\%(9/11)$ chloramphenicols that were EEC 4 plate positive or Charm II positive, but not detectrd by HPLC. Consequently, EEC 4-plate test method and Charm II test method were suitable by screening test on antibiotics and sulfonamides etc.

• Journal of Food Hygiene and Safety
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• v.11 no.4
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• pp.307-314
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• 1996

The European Economic Community four plate test(EEC 4-plate test, FPT, EU) has been used for monitoring antimicrobial drug residues in meat by Local Veterinary Service Center in Korea. This study was performed to evaluate sensitivity and group specificity of some antimicrobial drugs in FPT and to compare FPT with Charm II test. The minimal detectable levels of targeted antimicrobial drugs tested with standard solutions were 0.025∼1.0 ppm for 7 beta-lactams, 0.5∼1.0 ppm for 4 aminoglycosides, 0.05∼0.5 ppm for 5 macrolides, 0.05∼0.25 ppm for 3 tetracyclines and 0.25&1.0 ppm for 6 sulfonamides. In comparison of FPT and Charm II test, the results of FPT were not accord with those of Charm II test having the group specificity of seven families of antimicrobial drugs in meat samples except some families like tetracyclines.

• Korean Journal of Veterinary Service
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• v.20 no.2
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• pp.161-168
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• 1997

This study was undertaken to determined the sulfamethazine residues In urine, serum and muscle of slaughtered pigs(n: 230) by the method of TLC, EEC-4-plate and HPLC. 1. Inhibition diameter characteristics of reference bacterial substance by EEC-4-plate method and antibacterial inhibition zone was appeared sulfonamides in BS pH 7.2 2. Residual sulfamethazine was detceted from 7 serum(3.04%) and 10 urine(4.35%) by TLC test and concentration of residues was higher In urine than serum. 3. Residual sulfamethazine was detected from 9 muscle(3.91%) by EEC-4-plate method. 4. Positive samples by TLC test and EEC-4-plate method were exceed 0.1ppm quantitative analysis from 7 muscle(3.04%).

• Korean Journal of Veterinary Service
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• v.14 no.1
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• pp.63-69
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• 1991

Antibiotic residues of raw milks collected in Anyang area were tested by TTC-reduction test, EEC-4plate method and TLC(SOS-kit ) method to improve and monitor the quality of milk. Antibiotic substances were not detected from 100 raw milk samples, but sulfamethazine was detected from only one sample(1.3PPM) by SOS test. Unclassified anti-microbic substances were detected from 22 samples by EEC-4 plate method. EEC-4 plate test was useful to categorize the species of antibacterial substances and SOS test was useful to detect the sulfamethazine field screening test.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.3
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• pp.256-260
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• 1999

Minimun-detectable levels to 28 antibacterial agents used for the prevention and the treatment of fish diseases were determined to establish optimal detective method of bioassay in fish by the EEC 4-plate method, the modified method of EEC 4-plate and the standard method of analysis in food safety regulation. The test organisms used in the methods of bioassay were as follows: Bacillus subtilis BGA (B. subtilis) and Micrococcus luteus ATCC 9341 (M. luteus) in the EEC 4-plate method, B. subtilis, M. luteus and Bacillus cereus var. mycoides ATCC 11778 (B. cereus) in the modified of EEC 4-plate, and B. subtilis, M. luteus, B. cereus and Bacillus stearothermophilis var. calidolactis C-953 (B. stearothermophilis) in the standard method. The standard method showed predominant sensitivity in the detection of penicillins (PCs), and was also highly sensitive to aminoglycosides (AGs). The sensitivity of standard method in the detection of tetracyclines (TCs), marrolides (MLs), nitrofuran derivatives(NFs) and quinolones (QNs) was very low, and against sulfonamides (SAs), however, was extremely low. The modified method of EEC 4-plate showed very high sensitivity to TCs. Both the EEC 4-plate and the modified method of EEC 4-plate showed competitively high sensitivity in the detection of PCs, MLs, NFs, QNs and SAs. All the methods studied in the experiment showed very low sensitivity against chloramphenicol (CMs). Consequently, the modified method of EEC 4-Plate was the best bioassay method with a wide range of sensitivity for the optimal detection of the residual antibacterial agents in fish.

• Korean Journal of Veterinary Service
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• v.29 no.3
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• pp.331-338
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• 2006

This survey was carried out to detect the residual antibiotics in beef (n = 1,071), pork (n=7,837) and chicken (n=1,536) from slaughter houses in Gyeongbuk province by EEC-4 plate method, Charm II and HPLC during 2005. Residues of antibiotic were detected from 9 beef (0.8%) and 119pork (1.52%) by EEC-4 plate method, and total positive rates were 1.23% (128). 126 samples of the 128 positive samples by the EEC-4 plate method were detected by charm II test. 128 samples were classified as tetracyclines 110 (95.5%), ${\beta}$-lactam 2 (1.6%), sulfonamide 22 (17.2%), quinolone 1 (0.8%). The highest residual concentration of oxytetracycline, tetracycline, chlor-tetracycline, penicillin, sulfamethazine, sulfadimethoxine, sulfaquinoxaline, sulfamerazine, sulfamonomethoxine and enrofloxacin were 7.57, 0.27, 0.40, 0.24, 14.24, 4.33, 8.59, 0.12, 0.09 and 1.98 ppm, respectively and 49 samples were exceeded legal admitted levels.

• Korean Journal of Veterinary Service
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• v.20 no.2
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• pp.225-233
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• 1997

In an effort to Improve the quality of meat and to monitor farms, residual tetracyclines in local beef and pork produced in the province of North Chung-cheong were determined by a EEC 4-plate, Charm II and HPLC, respectively. The results were summarized as follows ; 1. Of the 547 samples, 4 beef and 13 pork samples were judged to be positive by EEC 4 - plate method. 2. Detection rates of tetracyclines by type 1(++--) and type 2(+++ or ++$\pm$-) micro-bial growth Inhibition In EEC 4-plate method were 100% and 71%, respectively. 3. Of 17 positive samples, 6 were positive for tetracyclines, 4 were positive for tetracyclines and sulfonamides, 1 was positive for sulfonamides, and 2 were positive for others by Charm II test. 4. The best eluents were 0.01M methanolic oxalic acid, and the ideal temperature for stable concentration was $40^{\circ}C$ as optimal HPLC analytical conditions for the detection of tetracy-clines. 5. Of the 10 positive samples for tetracyclines by Charm II test, tetracyclines were confirmed in 2 beef and 6 pork samples, using HPLC, at levels ranging from 3.64~4.22 ppm and 0.2~1.20 ppm, respectively.

• 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.

• 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.

• 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.