시료고체상분산(matrix solid phase dispersion)전처리법을 이용한 식육중 테트라사이클린계 항생물질 동시정량분석

강환구,손성완,조병훈,이혜숙,박신자,김재학,조명행,Kang, Hwan-goo,Son, Seong-wan,Cho, Byung-hoon,Lee, Hye-sook,Park, Shin-ja,Kim, Jae-hak,Cho, Myung-haing 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.3

Tetracycline antibiotics have been widely used not only therapeutics but feed additives. There are many methods for the isolation and determination of tetracycline antibiotics in animal muscle tissue. But those methods take much time and labor, so it is difficult to analyse many samples simultaneously. A rapid isolation method and liquid chromatographic determination of tetracycline antibiotics in animal muscle tissue (bovine, porcine, chicken) is presented. Blank control and tetracyclines fortified samples (0.5g) were blended with $C_{18}$ containing 0.05g each of oxalic acid and disodium ethylenediaminetetraacetate. After homogenize, homogenate was transferred to glass column made from 10ml glass syringe and compressed to 4~4.5ml volume. A column made from the $C_{18}$/meat matrix was washed with hexane (8ml) and dichloromethane (8ml, if needed), following which the tetracyclines were eluted,vith methanol or 0.01M methanolic oxalic acid (8ml). The eluates containing tetracyclines analytes were free from interfering compounds when analysed by HPLC with UV detection (photodiode array at 360nm). Standard curve for each tetracycline showed a linear response at the range of $0.05{\sim}1.0{\mu}g/ml$ and tetracycline antibiotics were eluted within 4ml of eluted volume. All tetracycline antibiotics except tetracycline were stable during the concentration process at $40^{\circ}C$ and time required for concentration was 3~4 hours. Fortified samples containing oxalic aicd and EDTA represented more good recoveries than those of not-contained sample. Recoveries were 91.8~110.1% (oxytetracycline; OTC), 57.7~79.5% (tetracycline; TC), 78.1~88.6% (chlortetracyclines; CTC) and 88.4~100.6% (doxycycline; DC) in pork tissue, 101.1~126.8% (OTC), 66.4~75.4% (TC), 79.2~88.1% (CTC) and 69.3~86.7% (DC) in beef tissue, and 90.8~95.6% (OTC), 66.2~84.4% (TC), 75.7~77.2% (CTC) and 55.6~80.7% (DC) in chicken muscle tissue. The detection limits validated in muscle tissue by this method were $0.05{\mu}g/g$ for OTC and TC, and $0.1{\mu}g/g$ for CTC and DC.

시료고체상분산(matrix solid phase dispersion)전처리법과 액체크로마토그라피를 이용한 돈육중 enrofloxacin 및 ciprofloxacin 분석

강환구,손성완,이혜숙,김재학,조명행,Kang, Hwan-goo,Son, Seong-wan,Lee, Hye-sook,Kim, Jae-hak,Cho, Myung-haing 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.1

A method for the isolation by matrix solid phase dispersion method and liquid chromatographic determination of enrofloxacin and ciprofloxacin in pork muscle tissue is presented. Blank or enrofloxacin and ciprofloxacin spiked samples(0.5g) containing 0.05g oxalic acid were blended with $C_{18}$(octadecylsilyl derivatized silica) packing material. After homogenization, $C_{18}$/muscle tissue matrix was transferred to glass column made from 10ml glass syringe and filter paper, and compressed to 4~4.5ml volume. A column was washed with 8ml of hexane and dried under vacuum. Interfering materials were removed by ethylacetate 8ml and dried, following which enrofloxacin and ciprofloxacin were eluted with 8ml of methanal under gravity. The eluate containing enrofloxacin and ciprofloxacin wase free from interfering compound when analysed by HPLC with UV detection at 278nm. Enrofloxacin and ciprofloxacin showed linear response with UV detector at the range of $0.05{\sim}1.0{\mu}g/ml$ and eluted within 5ml elution volume of methanol from the matrix. Fortified sample containing 0.05g oxalic acid represented more good recoveries than that of control sample. Average percentages of enrofloxacin and ciprofloxacin were $93.30{\pm}4.56%$ and $91.84{\pm}4.17%$, respectively, for the concentration range(0.05, 0.1, 0.25, 0.5 and $0.75{\mu}g/g$). The interassay variability of enrofloxacin was $6.02{\pm}5.33%$ with an intra-assay variability of 4.89% and $6.75{\pm}2.68%$ with 4.54% for ciprofloxacin. Detection limit of enrofloxacin and ciprofloxacin was $0.030{\mu}g/g$ in the spiked sample.

닭에서 ciprofloxacin의 체내 동태에 관한 연구

강환구,조명행,이항,한명국,손성완,김재학,이재진,Kang, Hwan-goo,Cho, Myung-haing,Lee, Hang,Han, Myung-guk,Son, Seong-wan,Kim, Jae-hak,Lee, Jae-jin 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.3

The purpose of this experiment was to develop a simple and reliable HPLC method for the detection of ciprofloxacin in chicken serum and to provide a basic data on pharmacokinetic parameters after oral and intramuscular administration. The results obtained were as follows: 1. 0.2% meta-phosphoric acid: acetonitrile(7:3, v/v) solution had a high and regular recovery rates and was selected as an extraction solution. 2. The recovery rates of ciprofloxacin were 83-97% with the selected solution in chicken serum and the detection limit was 50ng/ml in serum. 3. Ka(abosorption rate constant) were 3.652 1/h in fasted group and 0.880 1/h in non-fasted group, and Ke (elimination rate constant) were 0.061 1/h and 0.133 1/h, respectively. 4. The highest concentration in serum after intramuscular injection was 840ng/ml within 15-30min and 160-324ng/ml in 1.1-3.2 hours after oral administration. 5. The time course of blood concentration fits well into a 2 compartment model. 6. On oral administration of ciprofloxacin with feed, ciprofloxacin was absorbed more slowly and the amount of absorbed was smaller than that of in fasted chickens. 7. Blood concentration of ciprofloxacin increased in a dose-dependent manner after intramusclular and oral administraiton.

농약 잔류허용기준 면제 요건

강환구 ( Hwan Goo Kang ),문형준 ( Hyoung Joon Moon ),우계형 ( Gye Hyeong Woo ) 한국환경농학회 2021 한국환경농학회 학술대회집 Vol.2021 No.-

Some of Pesticides which are approved by pesticide control act could be exempt from the requirement of maximum residue level(MRL) in food by the regulation on the standards and specification of food in the Korean food code if they are applicable following condition. First they are not likely to pose risk to human because they have minimum risk. Second they are not likely to residue in food. Third, they are natural component of food and could not tell from exogenous compound. Fourth, natural plant protection product (including microorganism) which are proved to safe in human. We investigate the exemption regulation on pesticide MRL in United state of America. European Committee, Japan and Codex to suggest more detailed guideline which could be used when regulators determine the exemption of MRL for the pesticide in food items. In USA, the exemption for pesticide chemical residue in food are described in subpart D and subpart E in 40 CFR part 180 and 40 CFR 180.940 for the active/inert ingredients in antimicrobial products. The total amount of pesticide in/on the food items should not pose any harmful on the public health. Most of them are applicable when they are used under GAP and labeling. EU has the “guidance document on criteria for the inclusion of active substance into Annex IV (substances are not required the MRL) of regulation (EC) No 396/2005.(SANCO/11188/2013)”. They have 5 criteria for the exemption of MRL. Criterion 1 is the active substance which are approved as basic substance under regulation (EC) No 1107/2009. Criterion 2 is the compound is listed in Annex 1of regulation (EC) No 396/2005. Criterion 3 is the compound which has no identified hazardous properties. Criterion 4 is then natural exposure is higher than the one linked to the use of PPP. Criterion 5 is that no consumer exposure is forecasted linked to the mode of application of the PPP. establishment of codex maximum residue limit or do not raised to reside as step 4 from 2020”. The draft suggests 4 criteria. Criterion 1 is the basic substances and Active substances without hazardous properties identified (very low or no toxicological concern). Criterion 2 is substances for which it is not possible to differentiate between the exposure associated with its use as pesticide and its other uses in the food chain. Criterion 3 is the substances for which no consumer exposure linked to the mode of application is foreseen Criterion 4 is the microorganisms which are not pathogenic and do not produce mammalian toxins or other potentially toxic secondary metabolites of human health concern. Conclusively, most regulatory agency determined the exemption of pesticide MRL requirement during the approval procedure in the case of GAP and indication for use. The criteria include the risk factors (toxicity test data, experience of use), and the possibility of exposure to human (GAP, ADME, stability in the environment) and management option (residue analysis). This project was carried out by the fund from Ministry of Food Safety (211162농축산371) and we deeply appreciate it.

랫드에서 수은이 혈장 DNA와 혈액화학치에 미치는 영향

조준형,정상희,강환구,윤효인,Cho, Joon-Hyoung,Jeong, Sang-Hee,Kang, Hwan-Goo,Yun, Hyo-In 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.4

Changes of plasma DNA contents and serum biochemical values were measured in rats administered with $HgCl_2$ to investigate the in vivo cytotoxic effects of mercury and examine the usefulness of these changes as indicators of mercury exposure and diagnosis of mercury poisoning. Rats were given once intraperitonealy $HgCl_2$(0.13. 0.32. 0.8 and 2 mg/kg b.w) and the changes of plasma DNA contents and serum biochemical values were measured at the time of 2, 4, 8, 24, 48 and 72 hours after the administration of $HgCl_2$. Plasma DNA contents began to increase from 2 hours after the administration of $HgCl_2$ in all the treatment groups significantly compared to control with dose-dependent pattern. The levels of plasma DNA reached to peak at 48 hours as 2.77, 7.60, 15.46 and 16.51 times higher than control in each treatment group of 0.13, 0.32, 0.8 and 2 mg/kgb.w, respectively and remained to be higher until 72 hours after the administration. The values of creatine kinase, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, blood urea nitrogen and glucose of serum were increased, however the values of alkaline phosphatase, total protein and triglyceride were decreased. These changes of increase and decrease showed dose-dependent pattern but the starting time, maintenance and magnitude of change were various and characteristic according to serum biochemical indices. Among the changes of serum biochemical values, those of aspartate aminotransferase, lactate dehydrogenase and blood urea nitrogen were apparently and significantly increased compared to control from 2 to 72 hours by the administration of 2 mg/kg $HgCl_2$. This study demonstrates that plasma DNA and serum biochemical values such as aspartate aminotransferase, lactate dehydrogenase, blood urea nitrogen and etc. are valuable as biomarkers for mercury exposure assessment and diagnosis of mercury poisoning.

랫드에서 혈액검사에 영향을 미치는 요인들과 경정맥 채혈법의 시술에 관하여

조준형(Joon-Hyoung Cho),정상희(Sang-Hee Jeong),구현옥(Hyun-Ok Ku),강환구(Hwan-Goo Kang),박종명(Jong-Myung Park),윤효인(Hyo-In Yun),이영순(Yong-Soon Lee) 한국실험동물학회 1997 Laboratory Animal Research Vol.13 No.1

납의 생체내 세포독성 연구: 랫드에서 혈장 DNA와 혈액화학치 변화

조준형(Joon-Hyoung Cho),정상희(Sang-Hee Jeong),강환구(Hwan-Goo Kang),윤효인(Hyo-In Yun) 한국독성학회 2003 Toxicological Research Vol.19 No.3

Changes of plasma DNA contents and serum biochemical values were measured in rats administered with lead acetate to investigate the in vivo cytotoxic effects of lead and examine the usefulness of these in vivo cytotoxicity changes as indicators of lead exposure and diagnosis of lead poisoning. Rats were given once intraperitonealy with lead acetate (1.6, 8, 40 and 200 mg/kg b.w)<br/> and the changes of plasma DNA contents and serum biochemical values were measured at the time of 2, 4, 8, 24, 48 and 72 hours after the administration of lead acetate. Plasma DNA contents began to increase at 2 hours after the administration of lead acetate in the treatment groups of 8, 40 and 200 mg/kg b.w dose-dependently and significantly compared with control group. These DNA increases of each dosage group were continued until 24, 48 and 72 hours and the maximum levels of DNA (4.02, 10.67 and 14.10 times of control) were arrived at 8, 8 and 4 hours after the each treatment, respectively. Among 10 serum biochemical indicators, the activities of creatine kinase were increased to maximum level (6.55 times of control) at 2 hours after the administration and remained to be significantly higher than that of control by 8 hours in the treatment group of 200 mg, however, after 48 hours, the levels in the treatment groups of 40 mg above were lower than that of control. The values<br/> of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase were higher than that of control from 2 to 24 hours in the treatment group of 200 mg. Maximum levels of these enzymes were 3.34, 3.00 and 3.19 times of control, respectively. Both of alkaline phosphatase and triglyceride values in the treatment groups were decreased compared with control. In the case of alkaline phosphatse, the values were significanly decreased from 24 hours and more severely decreased until 72 hours in the treatment groups of 40 mg above (p<0.01). The minimum value was 0.36 times<br/> of control in the 200 mg group. The values of triglyceride were significantly decreased in the tratment groups of 40 mg above (p<0.01), but the values were not different significantly among the treatment groups. This study demonstrates that plasma DNA content and serum biochemical values such as aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase and triglyceride are valuable as biomarkers for exposure assessment and diagnosis of lead poisoning.

Androgenic and Antiandrogenic Activity of Butylated hydroxyanisole and Chlorpyrifos - methyl in Castrated Male Rats

(Sang Hee Jeong),(Dong Gyu Kim),(Joon Hyoung Cho),(Ok Kyung Kim),강환구(Hwan Goo Kang) 대한보건협회 2002 대한보건협회 보건종합학술대회 Vol.- No.-

Deoxynivalenol에 의한 생체독성 스크리닝 및 중독증 진단지표 확립

김은주(Eun-Joo Kim),정상희(Sang-Hee Jeong),구현옥(Hyun-Ok Ku),강환구(Hwan-Goo Kang),조준형(Joon-Hyoung Cho) 한국독성학회 2007 Toxicological Research Vol.23 No.2

Deoxynivalenol (DON) is a common foodborne mycotoxin and occurs predominantly in grains such as wheat, barley, oats, etc. DON induces systemic health problems such as loss of appetite, emesis and diarrhea in both human and farm animals. Reliable diagnostic parameters for DON intoxication are needed to prevent deep health impact. In order to establish useful diagnostic parameters, we investigated clinical signs, hematological values, serum biochemical values, gross-, histo- and toxico-pathological findings in B6C3F1 male mice after oral administration of DON (0.83, 2.5 and 7.5 ㎎/㎏) for 8 days. Body weight gain was significantly decreased at the highest dose of DON. Anorexia, ataxia, fur crudness and lack of vigor were observed at the highest dose DON group. In hematological values, the numbers of WBC and platelets and hemoglobin content were reduced with decreased neutrophil and monocytes by 7.5 ㎎/㎏ DON. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were prolonged in a dose-dependent manner and the content of fibrinogen was elevated at high dose of DON. Of serum biochemical values, total protein, globulin, BUN, cholesterol and testosterone were reduced but total bilirubin and albumin/globulin ratio increased. The enzyme activity of alkaline phosphatase was decreased while that of alanine aminotransferase was elevated. Relative organ weights of thymus, seminal vesicle/prostate and testes were dose-dependently reduced but those of liver and left adrenal gland increased with dose dependency. As for pathological findings, atrophy of thymus, seminal vesicle/prostate and testes and submucosal edema and ulceration in stomach and depletion of lymphocytes in thymus cortex were observed. In conclusion, these clinical, hematological, blood biochemical and patholgical parameters obtained in the present studies can be used for diagnosis of DON-mycotoxicosis, especially, low WBC, platelets, protein, BUN and testosterone and delayed prothrombin time can be available as for reliable diagnostic parameters.

랫드에서 Diazinon이 혈장 DNA와 혈액화학치에 미치는 영향

조준형 ( Joon Hyoung Cho ),정상희 ( Sang Hee Jeong ),강환구 ( Hwan Goo Kang ) 한국수의공중보건학회 2004 예방수의학회지 Vol.28 No.1