Endosulfan 과 그 분해산물의 Enzyme Immunoassay 에 의한 분석법의 개발과 응용

서용택,심재한,이강봉 ( Yong Tack Suh,Jae Han Shim,Kang Bong Lee ) 한국환경농학회 1992 한국환경농학회지 Vol.11 No.1

An enzyme immunoassay(EIA) was developed for the analysis of insecticide endosulfan and its degradation products. The sensitivity and specificity of the antibody produced were examined. Optimal conditions in the ELISA system for residue analysis were also discussed. A mixed suspension of endosulfan-hemocyanin conjugate(ES-KLH) 1.1 ㎎/㎖ and Freund`s adjuvant was injected subcutaneously to white rabbits and then collected antisera were tested for titers by indirect ELISA(1/24,000). Because of difficulties in the synthesis of endosulfan peroxidase conjugate, amine derivative of endosulfan-diol was synthesized and it showed 40% of conjugate yields(2㎎/㎖ of conjugate). the highest sensitivity obtained enzyme-conjugate was a concentration of 200ng/㎖. The detection limit of endosulfan in ELISA system was 5 ppb on the standard curve. In application of ELISA for residue analysis, the recoveries were really 100% both in the spiked soil and apple sample regardless of endosulfan concentration treated. On the other hand, chlorinated hydrocarbons of similar structure with endosulfan showed low cross-reactivity(2.2%∼29.2%).

사양토성 Lysimeter 에서 살균제 14C-Propiconazole 의 해방

김인선,서용택 ( In Seon Kim,Yong Tack Suh ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.3

Behaviour of a fungicide ^(14)C-propiconazole was investigate in a field lysimeter of sandy loam soil. At 15 days after rice-seeding transplanting, ^(14)C-propiconazole was treated on the soil surface at the rate of 0.12 ㎏/10a. The cummulative ^(14)C-propiconazole in the leachate from the lysimeter soil was 4.4% of the ^(14)C for 16 weeks. Most radioactivity detected in leachate was in the form of parent compound. At the end of lysimeter experiment. ^(14)C radioactivity in lysimeter soil was 76.5% of the applied ^(14)C and more than 97% of ^(14)C in soil remained in the top 20 ㎝. The percent of ^(14)CO₂ evolved from lysimeter soil was 7.8% of the applied ^(14)C. The radioactivity remained in the rice straw after harvest was 7.2% of the applied ^(14)C, while less than 0.1% of the applied ^(14)C was detected in flag leaf, ear, chaff and hulled rice, respectively.

사양토성 Lysimeter에서 살균제 $^{14}C-Propiconazole$의 행방

김인선,서용택,Kim, In-Seon,Suh, Yong-Tack 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.33 No.2

사양토성 lysimeter에서 살균제 $^{14}C-propiconazole$의 행방을 조사하고자 하였다. 벼 이앙 15일 후 $^{14}C-propiconazole$을 0.12 kg/10a 수준으로 lysimeter 토양표면에 처리한 후 시기별로 용탈수, 토양 및 휘발성분중 방사능을 조사하였다. 약제처리 후 16주간 lysimeter토양에서 용탈된 방사능은 처리 방사능의 4.4%였으며, 대부분의 방사능은 모화합된 형태이었다. 약제처리 16주 후 토양에 잔류하는 방사능은 처리 방사능의 76.5%였으며 97% 이상이 토양표층 깊이 20 cm 이내에 존재하였다. Lysimeter 토양에서 생성된 $^{14}CO_2$는 처리 방사능의 7.8%였다. 벼 수확 후 볏짚에 잔류하는 방사능은 처리량의 7.2%였으며 지엽, 벼이삭, 왕겨 그리고 현미에서는 각각 0.1% 이하를 나타내었다. Behaviour of a fungicide $^{14}C-propiconazole$ was investigated in a field lysimeter of sandy loam soil. At 15 days after rice-seedling transplanting, $^{14}C-propiconazole$ was treated on the soil surface at the rate of 0.12 kg/10a. The cummulative $^{14}C-radioactivity$ in the leachate from the lysimeter soil was 4.4% of the applied $^{14}C$ for 16 weeks. Most radioactivity detected in leachate was in the form of parent compound. At the end of lysimeter experiment. $^{14}C$ radioactivity in lysimeter soil was 76.5% of the applied $^{14}C$ and more than 97% of $^{14}C$ in soil remained in the top 20 cm. The percent of $^{14}CO_2$ evolved from lysimeter soil was 7.8% of the applied $^{14}C$. The radioactivity remained in the rice straw after harvest was 7.2% of the applied $^{14}C$, while less than 0.1% of the applied $^{14}C$ was detected in flag leaf, ear, chaff and hulled rice, respectively.

ELISA 기법을 이용한 농약의 잔류분석

이강봉,서용택 ( Kang Bong Lee,Yong Tack Suh ) 한국환경농학회 1993 한국환경농학회지 Vol.12 No.3

Immunochemical assay, ELISA for small molecules such as pesticides are rapid, sensitive, cost effective and can easily analyze with large samples. ELISA is one of several powerful biotechnologies immediately applicable to pesticide analysis. This review lists the advantages and disadvantages of the ELISA and elucidate the steps in assay development using examples from this laboratory. The focus is primarily on hapten synthesis strategies, protein conjugation, Immunization, assay format, and assay validation.

잉어중 Endosulfan 의 잔류분석을 위한 Radioimmunoassay ( RIA ) 의 개발

이강봉,심재한,서용택 ( Kang Bong Lee,Jae Han Shim,Yong Tack Suh ) 한국환경농학회 1994 한국환경농학회지 Vol.13 No.3

The established methods in the residue analysis of endosulfan require an extensive sample clean-up prior to quantification by relatively complex equipment. A radioimmunoassay(RIA) provides a simple procedure with theoretically higher sensitivity and specificity necessitating only a minimum of sample clean-up. Endosulfan-specific antibodies were developed in rabbits by using a bovine serum albumin(BSA) conjugate wherein the alcohol form of endosulfan was multiply bound to the protein via succinylation. Produced antibodies showed the high titers to endosulfan-BSA(1 : 32,000). An RIA method was developed in water and carp by using ^(14)C-labeled endosulfan as a tracer. The lowest detection amount of endosulfan was 1 ng in the liver, kidneys, gut and water samples, and 3 ng in the whole body sample of carp without any clean-up, corresponding to 0.1 ppb of endosulfan.

벼 재배 Lysimeter 토양중 살균제 14C - Propiconazole 의 행방

김인선(In Seon Kim),서용택(Yong Tack Suh) 한국환경농학회 1999 한국환경농학회지 Vol.18 No.3

Behaviour of a fungicide ^(14)C-propiconazole was investigated in a rice plant grown-lysimeter soil. The lysimeter was composed of soil cores of silty clay. Propiconazole(Tilt 250^R EC) plus ^(14)C-labeled propiconazole was applied on the surface of lysimeter soil at a rate of 0.12㎏/10a after rice transplanting. The application was done consecutively for two years. The behaviours of propiconazole in the lysimeter soil were investigated by measuring the amounts of ^(14)C-leachate, ^(14)CO₂, the residues distributed in each soil segment and taken up by rice plants. The relative amounts of ^(14)C leached from the lysimeter were the background level of the applied ^(14)C throughout expeiment. The amounts of ^(14)CO₂evolved from the lysimeter were 5.7 and 7.8% of the original ^(14)C in the Ist and 2nd treatment, respectively. The amounts of volatile substances soil were the background level throughout experiment, which indicated that propiconazole was stable chemically in the experimental condition. The ^(14)C-activities absorbed and translocated into rice plants were 3.7 and 7.6% in 1st and 2nd treatment., respectively. The ^(14)C-activities in the soil layer of the lysimeter was distributed mainly in the depth of 0 to 20㎝, which suggested propiconazole did not have the risk of groundwater contamination.

생쥐 체내에서 $^{14}C-{\alpha}-Endosulfan$의 대사

김인선,이강봉,심재한,서용택,Kim, In-Seon,Lee, Kang-Bong,Shim, Jae-Han,Suh, Yong-Tack 한국환경농학회 1996 한국환경농학회지 Vol.15 No.1

생쥐 체내에서 $^{14}C-{\alpha}-Endosulfan$의 흡수, 분포, 대사, 배설 등의 동태를 구명하기 위하여 약제를 7.5mg $kg^{-1}$ 수준으로 복강주사 처리하여 수행한 시험 결과를 요약하면 다음과 같다. 1. 약제처리 후 4 일 이내에 처리 방사능의 63.9%가 소변을 통해 배설되었고, 그 후부터는 배설율이 일정하였다. 2. 약제처리 후 각 조직별 방사능의 분포는 처리 30분 이내에는 심장에 가장 높게 분포하였으며, 2시간에는 간과 신장에서 그 분포율이 현저하게 증가하였으며, 시간이 경과함에 따라 그 분포율은 점점 감소하였다. 3. $^{14}C-{\alpha}-endosulfan$은 생쥐 체내에서 ${\beta}-endosulfan({\beta}-E)$, endosulfan ether(EE), endosulfan sulfate(ES), 그리고 endosulfan alcohol(EA)로 대사되었으며 주요 대사산물은 간에서 EA(13.25%) 그리고 신장에서는 EHE(19.37%)였다. 4. 소변중에 배설된 대사산물로는 EA(43.21%), ES(4.78%), ${\beta}-E$(7.21%), EE(3.72%), EHE(18.04%) 등이었다. Absorption, distribution, excretion and metabolism of $^{14}C-{\alpha}-Endosulfan$[1,4,5,6,7,7-hexachloro-8,9,10-=trinorborn-5-en 2,3-ylenebismethylene]sulfite) were studied in male mouse(Balb/c) after single intraperitoneal treatment as the dose level of 7.5 mg/kg body weights. After treatment of $^{14}C-{\alpha}-endosulfan$, the radioactivity was rapidly excreted into the urine(63.9 %) within 4 days, thereafter the excretion ratio was constant. Radioactivity levels in the tissues was reached maximum 0.5 hr in heart, 2 hrs in liver and kidney after the treatment, then decreased with time. Endosulfan was metabolized to ${\beta}-endosulfan({\beta}-E)$, endosulfan ether(EE), endosulfan sulfate(ES), and endosulfan alcohol(EA). The main metabolites were EA(13.25 %) in liver and endosulfan hydroxyether(EHE)(19.37 %) in kidney. The urinary metabolites were EA(43.21 %), ES(4.78 %), ${\beta}-E$(7.21 %), EE(3.72 %) and EHE(18.04 %).

생쥐 체내에서 14C - α - Endosulfan 의 대사

김인선,이강봉,심재한,서용택 ( In Seon Kim,Kang Bong Lee,Jae Han Shim,Yong Tack Suh ) 한국환경농학회 1996 한국환경농학회지 Vol.15 No.1

Absorption, distribution, excretion and metabolism of ^(14)C-α-endosulfan([1,4,5,6,7,7-hexachloro-8,9,10-=trinorborn-5-en 2,3-ylenebismethylene]sulfite) were studied in male mouse(Balb/c) after single intraperitoneal treatment as the dose level of 7.5 ㎎/㎏ body weights. After treatment of ^(14)C-α-endosulfan, the radioactivity was rapidly excreted into the urine(63.9 %) within 4 days, thereafter the excretion ratio was constant. Radioactivity levels in the tissues was reached maximum 0.5 hr in heart, 2 hrs in liver and kidney after the treatment, then decreased with time. Endosulfan was metabolized to β-endosulfan((β-E), endosulfan ether(EE), endosulfan sulfate(ES), and endosulfan alcohol(EA). The main metabolites were EA(13.25 %) in liver and endosulfan hydroxyether(EHE)(19.37 %) in kidney. The urinary metabolites were EA(43.21 %), ES(4.78 %), β-E(7.21 %), EE(3.72 %) and EHE(18.04 %).

Metalaxyl 의 다클론 항체생산과 최적 ELISA 조건

이강봉,임건재,정영호,서용택 ( Kang Bong Lee,Geon Jae Im,Young Ho Jung,Yong Tack Suh ) 한국환경농학회 1994 한국환경농학회지 Vol.13 No.1

A competitive indirect enzyme-linked immunosorbent assay(ELISA) was developed to detect and quantify levels of the fungicide metalaxyl in crops. Antiserum against metalaxyl was demonstrated in rabbits immunized with metalaxyl-human serum albumin(HSA) conjugate. Metalaxyl-protein conjugate was prepared by mixed anhydride and peptide coupling method with EDC. In this assay, metalaxyl-ovalbumin(OA) was coated(8㎍/㎖) on the microtiter plate, which was incubated for 1 hr at 4℃ or 4 hr at 37℃ with diluted antiserum(1:2,000). The optimum volume ratio of antigen and antibody mixture was 0.5: 1, which was incubated for 1 hr at 20℃. The detection of metalaxyl bound on the surface of wells was determined by the reaction(30 min) of antirabbit Ig G-peroxidase conjugate with its substrate.

작물체중 Metalaxyl 의 잔류분석을 위한 ELISA 의 응용

이강봉,임건재,정영호,서용택 ( Kang Bong Lee,Geon Jae Im,Young Ho Jung,Yong Tack Suh ) 한국환경농학회 1995 한국환경농학회지 Vol.14 No.1

The cross-reactivities of antibody derived from rabbit immunized with metalaxyl-HSA conjugate were identified to 44% and 28% for metalaxyl acid and metolachlor respectively. The detection range of metalaxyl in competitive ELISA was 5 ppb to 5 ppm. The recoveries of metalaxyl in ELISA for 6 crops; potato, sesame, pepper, cabbage, cucumber and onion was ranged 77.5-103.6%. In this test, CV% were calculated below 10% except for sesame sample as shown 21.5%.