A screening method has been developed for detecting sulfamethazine(SMZ) contamination of meat or feeds by using horseradish peroxidase (HRP) labeled protein A (Prot AHRP)and an indirect competitve enzyme-linked immunosorbent assay(ELISA). The assay is based on competitve binding of guinea pig anti-SMZ with SMZ in smaple and SMZ-gelatin conjugate(SMZ.GEL). Percent binding (B.Bo$\times$100) was calculated from the absorbance in the absence (B0) and presence (B) of SMZ. By the sandard curve prepared by plotting log(SMZ) vs percent binding of each known reference solution, the detection limit was 1.0ppb or less. Cross reacton with sulfadimethoxine, sulfaguaniding, sulfamerazine, sulfamthoxpyridazine, sulfanilamide, sulfisomidine and sufisoxazole were not observed. But sulfamerazine crossreacted in the test. The EC-50 value (concentration causing 50% inhibition of color development compared with blank) of sulfamerazine was 2.0 ppm. Further quality control will make the ELISA system ideal for the detection of SMZ in meat or feeds.
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.
The sulfonamide is one of potentiative antimicrobial agents which is being used widely in veterinary medicine for control of several animal diseases such as mastitis as well as for promotion of growth. However, the misusages of sulfonamides in food producing animals, especially cattle produce several considerable problems in human health caused from residues of this antibiotic in milk product. To determine the most effective analytical methods for residual sulfonamides in raw milk, this study was performed comparatively using by some applicable screening detecting method such as TTC, Charm II test (sulfonamides), and Lactek tests (sulfamethazine kit). The positive result from screening tests was confirmed by HPLC method. Milk samples (540 raw milks) were collected from dairy farms. Results of this study are summariezed as follorrs ; 1. All samples (540 raw milks) showed negative response from TTC test, however, 18 raw milks of those samples responded positively to Charm II test. 2. By Lactek test, residual sulfamethazine was detected from 4 raw milks. Fifteen raw milks of 18 samples which were classified as positive one by Charm II test, showed positive response 3. Retention time of sulfonamides added at the level of 100ppb into sklm milk was ranged from 1.55 minute to 23.3 minute. Recovery rates of sulfonamides were variable from 6.7% upto 94.2% depended on the types of sulfonamlde. 4. Single type of sulfonamides was detected from 10 raw milk samples, 2 types of sulfonamides from 3 samples and 3 types from 2 raw milks by HPLC. 5. Sulfonamides was detected in this study were 5 types : 11 samples for sulfisomidine, 5 samples for sulfamethazine, 3 samples for sulfadlmethoxine, 2 samples for sulfathiazole and 1 sample for sulfadiazine. 6. The highest levels of residual sulfonamides was 210.3 ppb of sulfamethazine but the lowest concentration of residue was 2.2 ppb of sulfamethazine and sulfisomidine, respectively. Number of samples detected positively in this experiment were belows : above 100 ppb for 1 sample (4.5%) (sulfamethazine), 50~100 ppb for 4 samples (18.1%) (each 2 samples for sulfamethazine and sulfisomidine, respectively), 25~50 ppb for 6 samples (27.1%) (2 sulfisomidine, each 1 sample for sulfadiazine, sulfadimethoxine, sulfamethazine and sulfathiazole, respectively), 10~25ppb for 3 samples (13.7%) (3 sulfisomidine), and below 10ppb for 8 samples (36.4%) (4 sulfisomidine, 2 sulfadimethoxine and each 1 for sulfamethazine and sulfathiazole).
Park, J.T.;Jeong, E.J.;Kim, Y.G.;Song, B.J.;Oh, K.S.;Lim, H.C.;Kim, S.C.
Korean Journal of Food Science and Technology
/
v.26
no.3
/
pp.221-225
/
1994
This survey was carried out to determine five sulfonamide(sulfamerazine, sulfamethazine, sulfadimethoxine, sulfamonomethoxine, sulfaquinoxaline) residues in beef, pork, chicken and swine kidney. For this survey, 30 samples of beef, 15 samples of chicken, 10 samples of pork and 10 samples of swine kidney were collected in Chonnam from June, 1992 to June, 1993, and were analyzed by HPLC. The recoveries of sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfadimethoxine, and sulfaquinoxaline in spiked samples between $0.25{\sim}1.00$ ppm were 71.7%, 80.3%, 71.6%, 70.9%, 68.4%, respectively. None of 65 samples which were examined exceeded 0.1 ppm. Of 15 chicken muscle samples, 2 samples exceeded 0.05 ppm in sulfamerazine (0.077 ppm) and sulfamethazine (0.075 ppm), respectively. Of 10 swine kidney samples, 1 sample exceeded 0.05 ppm in sulfadimethoxine (0.052 ppm). And sulfanilamide concentration of swine kidney were higher than pork.
This experiment was carried out to detect the residues of sulfonamides in raw milk. Raw milks which does not contain sulfonamides was collected from one of the farm and fortified with 5 sulfonamides (sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfadimethoxine, sulfaqinoxaline). The sulfonamides in the fortified sample were extracted and detected by High Performance Liquid Chromatography. UV /vis detector was used in this experiment. The results obtained were summarized as follows : 1. Chloroform was good as a extracting solution. 2. 15.5% methanol in PDP as a mobile phase solution was best detective condition for SMR, SMT, SMM. But for SDM and SQN the best condition was 23% methanol. 3. The detectable limits of SMR, SMT, SMM were 2ppb. but SDM and SQN were 20ppb because of delayed retention time and relatively low recovery rate. 4. The peaks of SMR, SMT, SMM and SDM were erected at baseline and the apexes were sharp but SQN was round shape.
Journal of agricultural medicine and community health
/
v.18
no.1
/
pp.55-63
/
1993
A simultaneous determination method by HPLC for egg-residues sulfamerazine, sulfamethazine, sulfadimethoxine, furazolidone and zoalene was assesed. The drugs were extracted by dechloromethane, The extract after solvent evaporation, is partitioning in hex ane/water and back-partitioning in dechloromethane and analysis by HPLC. The average recovery rates of the above microbials from the egg spiked standard solution were approximately 81.2%, 87.6%, 92.5%. 86.1% and 79.3% respectively. The limit of detection of sulfamerazine. sulfamethazine and sulfamethoxine were in the levels of 0.2ppb, furazolidone and zoalene 0.5ppb respectively. According to this method 84 commercial eggs were examined. Sulfamethanzine was detected at levels of 0.005-0.008ppm in 3 eggs. Sulfadimethoxine was detected at levels of 0.012-0.019ppm in 4 eggs. No sulfamerazine, furazolidone and zoalene was detected in every samples. The residues of antimicrobial agent were safety level as food generally.
Kang Hee-Joo;Choi Kyung-Ho;Kim Min-Young;Kim Pan-Gyi
Journal of Environmental Health Sciences
/
v.32
no.3
/
pp.227-234
/
2006
There has been increasing concern in recent years about the occurrence, fate and toxicity of pharmaceutical products in aquatic environment. Although these compounds have been detected in a wide variety of environmental samples including sewage effluent, surface waters, ground water and drinking water, their concentrations generally range from the low ppt to ppb levels. It is therefore often thought to be unlikely that pharmaceuticals will have a detrimental effect on the environment. This study was conducted to determine the endocrine disruption effects of the several pharmaceutical residues in water using adult Japanese medaka (Oryzias latipes). The common antibiotics were used sulfa durgs (sulfamethoxazole and sulfamethazine) and tetracycline drugs (oxytetracycline and tetracycline). Positive control that was induced Vtg (vitellogenin) in male fish was used $17\beta$-estradiol. Vtg was qualified and quantified through Western blotting and ELISA. After SDS gel electrophoresis, the dominant protein band was identified to molecular weight approximately 205 kDa in whole body samples of vitellogenic female. In female medaka exposed to $17\beta$estradiol, there was no significant difference in total protein induction. In contrast, three to five day exposure of male fish to $17\beta$-estradiol resulted in more than 60.0% increase of total protein compared to that of control males (p<0.01). In case of antibiotics, female fish didn't show significant difference, but male fish was showed significant difference. In addition, Vtg induction in male fish was observed with all the test chemicals. On concentrations greater than 0.1 ppm of sulfamethoxazole, 1 ppm of sulfamethazine, 1 ppm of oxytetracycline and 20 ppm of tetracycline, Vtg induction was increased in a dose response manner. This study is one of the early reports suggesting potential endocrine disruption mechanism of antibiotic pharmaceutical products in aquatic ecosystem. Although the effect concentrations obtained from this study were high as unrealistically as in environments, it is endocrine disruption that we should be considered as one of the important consequences of pharmaceutical contamination at water environment, and warrants due attention in future researches.
The antibiotic residues of the urine, the liver, the lung, the kidney and the spleen in slaughtered pigs at Kyongbuk province were detected by TLC(505 kit) and EEC-4 plate method. 1. The positive rate of residual sulfamethazine which was detected by 505 kit in the urine (n=200) was 0.0%. 2. The positive rate of residual sulfamethazine which was detected by EEC-4 plate in the urine (n=126), the liver(n=98), the kidney(n=72), the spleen (n=68) and the lung(n=48) were 63%, 49%, 36%, 34% and 24%, respectively. 3. By EEC-4 plate method, the positive detection rates of the urine were 53.0% in BS(pH 6.0), 29.0% in BS(pH 7.2), 11.5% in BS(pH 8.0) and 13.0% in ML(pH 8.0) medium, that of the liver 41.5% in BS(pH 6.0), 22.0% in BS(pH 7.2), 6.5% in BS(pH 8.0) and 5.0%, in ML (pH 8.0) medium, that of the lung 21.0% in BS(pH 6.0), 9.5%, in BS(pH 7.2) and 8.5% in BS(pH 8.0) medium, and that of the kidney 31.5% in BS(pH 6.0), 14.5% in BS(pH 7.2), 20.0% in BS(pH 8.0) and 3.0% in ML(pH 8.0) medium. In the spleen, only in BS(pH 6.0) medium the positive rate was detected as 33.5 %. 4. The positive rates of samples which shown TLC-positive detected by EEC-4 plate method were 53.9% in no band, 77.8% in one band, 80.9% in two bands, 66.7% in three bands, respectively. In conclusion, the EEC-4 plate method could be applied for the detection of residual antibiotics in samples which shown as out of standard Rf values by TLC-method (SOS kit).
In order to know the depletive changes of sulfadimethoxine and oxytetracycline residues in se겨m of dairy cattle intramusculally administered with sulfadimethoxine sodium(SDS) and oxytetracycline(OTC), the concentration of sulfadimethoxine and oxyteracycline was measured in serum of dairy cattle with using high performance liquid chromatography(HPLC). SDS and OTC was intramuscularlly administrated to dairy cattle at the rate of 10mg/kg(SDS) and 10mg/kg(OTC) body weight(recommended therapeutic dose) once to four dairy cattle. There were investigated the depletive changes of the sulfadimethoxine and oxytetracycline in serum of dairy cattle at the time 2, 4, 8 hours, 1st, 2nd, 3rd, 4th and 5th day after administration SMS and OTC, respectively. The results obtained were summarized as follows; 1. After intramuscularlly administration of the SMS, the mean concentrations of sulfamethazine in serum according to the time lapsed were showed 33.964 $\pm$ 4.435ppm at the 4 hours after withdrawal of medicated sulfadimethoxine sodium. And gradually according to the time lapsed, the concentrations of sulfadimethoxine residues in serum were significantly (p<.05) decreased 6.596 $\pm$ 3.402 ppm at 1st day, 0.217 $\pm$ 0.119 ppm at 3rd day and 0.057 $\pm$ 0.032 ppm at 4th day, respectively. 2. The mean residual concentration of OTC in serum according to the time lapsed after intramuscularly administration OTC were showed 0.743 $\pm$ 0.368ppm at the 8 hours. And gradually according to the time lapsed, the mean concentrations of OTC residues in serum of dairy cattle were significantly(p<.05) decreased such as 0.057 $\pm$ 0.047ppm at 3rd day and 0.039 $\pm$ 0.016ppm at the 5th day, respectively. In conclusion, this study could be suggested the relationship between administrated period, sulfonamides and tetracycline residual aspects in serum, and the importance of observing ceasing period of antibiotic drugs before forwarding livestocks to slaughter, Thus, this results would be able to be used the basic index for prevention of sulfonamides and tetracycline residues in dairy breedings.
A simple, rapid and simultaneous analytical method is described for the detection of Sulfonamide and Tetracycline residues, i.e., Sulfamerazine (SMR), Sulfamethazine (SMT), Sulfamonomethoxine (SMM), Sulfadimethoxine (SDM), Sulfaquinoxaline (SQN), Oxytetracycline (OXY), Tetracycline (TC), Chlortetracycline (CTC). Blank control and sulfonamide and tetracycline fortified fish muscle samples (0.5 f) were blended with octadecylsilyl (C18, 40 ${\mu}{\textrm}{m}$, 21% load, 60$\AA$) derivatized silica packing material (2 g). Blended fish samples were washed with hexane, then, benzene and dichloromethace were used for the elution of tetracycline and sulfonamide were analyzed by HPLC. Correlation coefficients of standard curves for individual sulfonamide and tetracycline isolated from fortified samples were linear (0.9993$\pm$0.0003~0.9997$\pm$0.0003, 0.9493$\pm$0.078~0.9753$\pm$0.036), respectively. The average percentage recoveries of sulfonamide and tetracycline ranged as 80.86~96.52% to 85.88~92.23%, and 30.01~37.12% to 65.89~73.40%, for the concentration range (0.1~1.0 ppm) examined, respectively. Limit of detection for sulfonamide was 0.0012 ppm for SMR in Paralichthys Olivacleus and 0.0020 ppm for SMR, 0.015 ppm for SMM in Cyprinus Carpio. The applicability of this procedure is demonstrated by separation and detection of incurred tetracycline and sulfonamide residues in fish muscle tissue.
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