• 제목/요약/키워드: Antibiotic residues

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LC-MS/MS를 이용한 농산물 중 Kasugamycin 시험법 개발 (Development of Analytical Method for Kasugamycin in Agricultural Products using LC-MS/MS)

  • 이한솔;도정아;박지수;조성민;신혜선;장동은;정용현;이강봉
    • 한국식품위생안전성학회지
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    • 제34권3호
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    • pp.235-241
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    • 2019
  • 본 연구는 농산물 중 잔류허용기준 신설 예정 농약인 가스가마이신의 안전 관리를 위한 공정 시험법을 개발하기 위하여 수행되었다. 가스가마이신은 단백질 합성과정 중 번역 개시 단계를 저해하여 균의 증식을 억제하는 작용기작을 가지고 있으며 특히 사상균인 Piricularia oryzae에 의해 발병하는 도열병을 예방하는 데 효과적이다. 현재 우리나라에는 농산물 중 가스가마이신의 잔류물의 정의가 설정되어 있지 않고 모화합물 만을 잔류물의 정의로 예정하고 있다. 또한 잔류허용기준이 설정된 농산물이 없기 때문에 모든 국내 재배 농산물 또는 수입 농산물 유통 시 실시하는 잔류농약 검사에서 농약 허용물질목록관리제도(Positive List System; PLS)에 의하여 잔류량이 0.01 mg/kg 이하가 되어야 한다. 이에 농산물 중 잔류농약 분석 및 검사를 위한 공정 시험법 마련이 시급하여 본 연구에서는 대표농산물 5종(현미, 감자, 대두, 감귤, 고추)을 대상으로 시험법을 개발하고자 하였다. 따라서 수용성 유기용매인 메탄올의 적용과 수산화나트륨을 이용한 pH 조절을 통한 추출법 및 HLB 카트리지를 이용한 정제법을 최적화하여 LC-MS/MS에 의한 분석법을 확립하였다. 가스가 마이신의 시험법 정량한계는 0.01 mg/kg이며 5종의 농산물에 0.01, 0.1 및 0.5 mg/kg의 처리농도로 회수율 실험을 한 결과 평균 회수율은 71.2~95.4%이었고, 상대표준 편차는 12.1% 이하로 조사되었다. 이러한 검증 결과는 국제식품규격위원회 가이드라인(CAC/GL 40-1993, 2003)의 잔류농약 분석 기준 및 식품의약품안전평가원의 '식품등 시험법 마련 표준절차에 관한 가이드라인(2016)'에 적합한 수준임을 확인하였다. 따라서 본 연구에서 개발한 시험법은 국내 및 수입 농산물 중 가스가마이신의 안전 관리를 위한 공정시험법으로 활용될 수 있으며 잔류물의 정의 및 잔류허용기준을 설정하는 데 기초자료로써 활용 가능할 것이다.

Effects of Substitutions of Gln16 and Asp18 with Phe or Tyr in HP(2-20) on its Structure and Antimicrobial Activity

  • Kim, Jin-Kyoung;Lee, Ju-Un;Kim, Woong-Hee;Park, Yoon-Kyung;Hahm, Kyung-Soo;Kim, Yang-Mee
    • 한국자기공명학회논문지
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    • 제13권1호
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    • pp.27-34
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    • 2009
  • HP (2-20), a 19-residue peptide derived from the N-terminus of Helicobacter pylori Ribosomal Protein L1, has antimicrobial activity but is not cytotoxic to human erythrocytes. Previously, we have synthesized several analogue peptides to investigate the effects of substitutions on the structure and antimicrobial activity. Substitution of $Gln^{16}$ and $Asp^{18}$ with Trp (Anal 3) caused a dramatic increase in bacterial and fungal lytic activities. In this study, analogue peptides were synthesized to investigate the effects of substitution of Gin and Asp with Phe (Anal 6) or Tyr (Anal 7) in HP (2-20) on its structure and antimicrobial activity. Substitution of Gin and Asp with hydrophobic aromatic residues at position 16 and 18 of HP (2-20) caused increase in antibiotic activity without hemolytic effect. Substitution of Gin and Asp with Trp and Try increased antibiotic activity of HP (220) twice more compared to substitution with Phe. The tertiary structures of Anal 6 and Anal 7 in SDS micelles has been investigated using NMR spectroscopy. The structures revealed that substitutions of the aromatic residues at C-terminus resulted in longer and well defined alpha-helix and improved their antibacterial activities

시료 중 잔류 항생제 분석 방법: I. 고상 시료 전처리 방법 (Determination of Antibiotic Residues: I. Extraction and Clean-up Methods for Solid Samples_A Review)

  • 김찬식;류홍덕;정유진;김용석;류덕희
    • 한국물환경학회지
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    • 제32권6호
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    • pp.600-627
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    • 2016
  • Korea is one of the countries with a large veterinary antibiotics market, although antimicrobial resistance in bacteria is becoming a serious issue in many countries. The Korean government started to take interest in estimating the effects of livestock manure on rivers and agricultural soils and in monitoring of heavy metals, organic pollutants and antibiotics in the ambient water and soil. In this paper, pre-treatment methods to separate the selected antibiotics from solid samples were reviewed. It is essential to select an efficient and appropriate procedure for pre-treatment due to the high proportion of proteins and organics in biosolid samples. Pre-treatment consists of extraction followed by clean-up. Initially, homogenized samples were extracted by sonication, mechanical agitation or pressurized liquid extraction with methanol/acetonitrile/water mixture under acidic/basic conditions depending on the compound. However, aminoglycosides and colistin were extracted with 5% trichloroacetic acid and HCl, respectively. Since the ${\beta}-lactams$ are easily decomposed in acidic and basic conditions, they were extracted in neutral pH. Filtration with a membrane (pore size, $0.2{\mu}m$) or solid phase extraction with HLB and methanol, as eluents, was normally applied for the clean-up. At least, three different pre-treatment procedures should be adopted to screen all the selected antibiotics in solid samples.

광주지역 생식용 소 부산물의 위생관리 실태 조사 (A survey on hygiene management for raw by-products of beef in Gwangju area, Korea)

  • 김지연;장미선;고바라다;지태경;성창민;박다해;김현중;김은선;김용환
    • 한국동물위생학회지
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    • 제36권3호
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    • pp.209-216
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    • 2013
  • A total of 301 samples of bovine liver, spleen and omasum were collected from butchers and restaurants in Gwangju, Korea during 2012 and all samples were subjected to bacteriological examination and antibiotic residues. Also, this study was performed to survey the consciousness for hygiene of livestock workers who are handling bovine by-products in Gwangju. The detection rate of aerobic plate count (APC) was higher in summer than in other seasons in all by-products (P=0.000). The detection rate of E. coli count was lower in the liver than the spleen and omasum (P=0.000). Twenty four of the samples (8.0%) were contaminated with S. aureus while one spleen sample (0.3%) was contaminated with L. monocytogenes and finally 10 (3.3%) of the liver and omasum samples were contaminated with Cl. perfringens. Five of the twenty-four S. aureus isolates harbored enterotoxin gene. However, the cpe gene of Cl. perfingens was not detected among any of the 10 isolates. Antibiotic residues were not detected in the liver samples. The consciousness survey's results showed that most of them (58.8%) were safe.

유우에 oxytetracycline 및 sulfadimethoxine sodium 투여 후 혈청내 잔류함량 변화추이 (Changes of residual concentration in serum of the dairy cattle after administration of oxytetracycline and sulfadimethoxine sodium)

  • 도재철
    • 한국동물위생학회지
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    • 제25권2호
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    • pp.141-151
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    • 2002
  • 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.

절박 도축우의 항생제 및 설파제 잔류 조사 (Detection of residual antibiotic and sulfonamide agents in emergency slaughtered cattle)

  • 백미순;이영철;이해영;박병옥;유기승;조중현;박유순
    • 한국동물위생학회지
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    • 제21권1호
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    • pp.13-20
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    • 1998
  • This survey was carried out to detect the residual antibiotics and sulfonamides in emergency slaughtered cattle(n=265) from slaughter houses in Kyeonggi province by EEC 4-plates method, Charm II and HPLC. The results were summarized as follows ; 1. Antimicrobial substances were detected from 24 samples(9.1%) by EEC 4-plates method and the detection ratio were highest in summer(13.8%). 2. Twenty-two of the 24 positive samples were classified as sulfonamide(34.4%), TCs(31.3%), $\beta$-lactam(23.5%) and aminoglycoside(9.3%) by Charm II test. 13(59.1%) of the 22 samples contained single agent and 9 samples(40.9%) contained 2 or more agents. 3. Oxytetracycline(27.3%), penicillin G (27.3%) and sulfathiazole(18.2%) were detected in 20 from 22 samples by the HPLC and Charm II test. 4. The residual concentration of oxytetracycline, penicillin G and sulfonamide were 0.29~9.30 ppm, 0.05~9.58 ppm and 0.04~7.59 ppm, respectively and 19 samples(7.2%) were exceeded tolerance levels.

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Synthesis and Characterization of GGN4 and its Tryptophan Substituted Analogue Peptides

  • Kim, Se-Ha;Kim, Ji-Young;Lee, Byeong-Jae;Kim, Soon-Jong
    • BMB Reports
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    • 제32권1호
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    • pp.12-19
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    • 1999
  • Gaegurin 4 (GGN4), a broad-spectrum antibiotic, is a 37-amino acid peptide isolated from the Korean frog, Rana rugosa. In this study, we have chemically synthesized and purified GGN4 analogues where the C-terminal portion is truncated and/or substituted with tryptophan. These peptides show significantly different biological activities depending on the location of tryptophan and the number of amino acids truncated from the C-terminal end. While deletion of 9 amino acids from the C-terminal seems to be marginally tolerable in maintaining the antimicrobial activity, further deletion of up to 14 amino acid residues decreases the potency by more than 60-fold towards Gram-positive, and 10-fold towards Gram-negative, bacteria. Surprisingly, the reduced activity of the shorter peptide can be completely restored by a single substitution of aspartic acid 16 to tryptophan 16 (D16W). Also, the truncation seems to decrease the specificity of antibiotic activity more towards Gram-positive than towards Gram-negative bacteria studied. These data suggest a partial role of the C-terminal region in determining the binding specificity and the activity of peptides upon binding to their target cell membranes.

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육류중 잔류 항생물질 및 테트라싸이클린 조사 (Detection of Residual antibiotics and tetracyclines in beef, pork and chicken)

  • 백미순;이영철;심항섭;박병옥;조중현;박유순
    • 한국동물위생학회지
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    • 제20권4호
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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.

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의약물질의 환경오염과 환경보건 (Pharmaceuticals in Environment and Their Implication in Environmental Health)

  • 최경호;김판기;박정임
    • 한국환경보건학회지
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    • 제35권6호
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    • pp.433-446
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    • 2009
  • Pharmaceuticals in the aquatic environment are trace contaminants of growing importance in environmental health due to their physiologically active nature. Pharmaceuticals could affect non-target species and might eventually damage sustainability of susceptible populations in the ecosystem. Potentials for health consequences among susceptible human population cannot be ruled out since long-term exposure to cocktails of pharmaceuticals, which might be present in drinking water, is possible. Selection of antibiotic resistant microorganisms is of another concern. In order to understand, and if needed, to properly address the environmental health issues of pharmaceutical residues, knowledge gaps need to be filled. Knowledge gaps exist in many important areas such as prioritization of target pharmaceuticals for further risk studies, occurrence patterns in different environments, chronic toxicities, and toxicities of pharmaceutical mixtures. Appropriate treatment technologies for drinking water and wastewater could be developed when they are deemed necessary. One of the simplest, yet most efficient measures that could be undertaken is to implement a return program for unused or expired drugs. In addition, implementation of environmental risk assessment frameworks for pharmaceuticals would make it possible to efficiently manage potential environmental health problems associated with pharmaceutical residues in the environment.

위치 지정 치환 변이를 이용한 ErmSF의 '타깃 Adenine Binding Loop'을 형성하는 부위에 존재하는 223/227 Arginine 잔기의 23S rRNA Methylation 활성에서의 역할 규명 (Site-directed Mutagenesis Analysis Elucidates the Role of 223/227 Arginine in 23S rRNA Methylation, Which Is in 'Target Adenine Binding Loop' Region of ErmSF)

  • 진형종
    • 미생물학회지
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    • 제48권2호
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    • pp.79-86
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    • 2012
  • ErmSF는 23S rRNA의 A2058 (E. coli numbering)에 methylation을 유발하여 macrolide-lincosamide-streptogramin B ($MLS_B$)계 항생제의 부착을 저해함으로써 항생제 활성을 억제하는 내성인자 단백질인 Erm 단백질들 중의 하나이다. Erm 단백질들 사이에서 공통적으로 나타나는 $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) 서열은 Erm 단백질인 ErmC'와 DNA methyltransferase인 M. Taq I의 구조를 분석한 연구에서 타깃인 adenine과 직접적으로 상호작용하는 부위로 제안되거나 확인되었다. 따라서 이 부분 중 Erm 단백질 사이에서 잘 보존되어있지는 않지만 염기성인 잔기의 특성상 기질인 RNA와 상호작용이 예상되는 223, 227번 arginine을 alanine으로 위치 지정 치환한 변이 단백질을 이용하여 그 잔기의 효소 활성에서의 역할을 확인하였다. 두 변이 단백질은 생체 내에서 그 활성을 여전히 유지하고 있어서 항생제인 erythromycin에 대하여 내성을 나타내었으나 in vitro 상에서는 R223A 또는 R227A가 야생형 ErmSF에 비하여 약 50%, 88%의 활성을 각각 나타내어 효소 활성에서 각 잔기가 결정적이지는 않지만 중요한 역할을 수행하고 있음을 확인하였다.