J Korean Soc Food Sci Nutr 한국식품영양과학회지 44(9), 1286~1294(215) http://dx.doi.org/1.3746/jkfn.215.44.9.1286 감마선조사오렌지의급성및아만성독성평가 정다운 1 황옥화 1 최근표 2 강일준 1 1 한림대학교식품영양학과 한국영양연구소 2 강원도립대학식품가공제과제빵과 Acute and Subchronic Toxicity of Gamma-Irradiated Orange Da-Woon Jung 1, Yu-Hua Huang 1, Geun-Pyo Choi 2, and Il-Jun Kang 1 1 Department of Food Science and Nutrition & The Korean Institute for Nutrition, Hallym University 2 Department of Food Processing and Bakery, Gangwon Provincial College ABSTRACT The acute and subchronic toxicity of gamma-irradiated orange was evaluated in ICR mice. For acute toxicity, groups of 3 male and 3 female ICR mice were orally administered gamma-irradiated orange (, 1,, and 2, mg/kg). The mortality, clinical sign, body weight changes, and necropsy findings of ICR mice were observed for 14 days. No significant changes in body weight or abnormal gross findings were observed in relation to gamma-irradiated orange. Hematological and serum biochemical parameters were within normal ranges. According to the results, gamma-irradiated orange had no special toxic effects in male and female ICR mice at 2, mg/kg. For subchronic toxicity, groups of 36 male and 36 female ICR mice were given a diet of gamma-irradiated orange for 13 weeks (control, non-irradiated, and irradiated imported orange). During the experimental period, mortality, clinical signs, body weight change, food consumption, organ weight, and histopathological examination did not show any changes in comparison to the control group. Several hematological and serum biochemical parameters showed statistically significant changes, but these changes were within normal range. These results indicate that 1 kgy gamma-irradiated orange did not cause any toxic effects in male and female ICR mice and therefore can be considered as safe. Key words: acute toxicity, subchronic toxicity, gamma-irradiation, orange, safety 서 오렌지는감귤류에속하는열매의하나로 beta-carotene, vitamin C, flavonoid 등의생리활성물질이많이함유되어있어우리나라를포함해전세계에서널리소비되고있는과일이다 (1). 우리나라에서소비되는오렌지의대부분은미국에서수입되고있으며, 매년그양도증가하고있다 (2). 과실류의국제교역과유통등에서저장기간을연장하고미생물과해충감염을방지하기위한방법으로 ethylene oxide(eo) 와 methyl bromide(mebr) 를이용한훈증살균기술이일반적으로사용되고있다 (3). 그러나 EO는발암성과잔류성등의문제점이보고되어국제적으로식품에사용을제한하였고, MeBr도오존층을파괴하는환경공해물질로규명되어사용을제한하였다 (4). 그밖에보편적으로사 Received 11 May 215; Accepted 6 August 215 Corresponding author: Il-Jun Kang, Department of Food Science and Nutrition & The Korean Institute for Nutrition, Hallym University, Chuncheon, Gangwon 24252, Korea E-mail: ijkang@hallym.ac.kr, Phone: +82-33-248-2135 론 용하고있는살균처리에는가열처리, 자외선조사, 화학약품처리등이있지만, 식품성분의변화및손실을가져오거나화학성분의잔류및유해물질을생성하는등에의한문제점으로인해효과적인대체기술개발이필요하다 (5,6). 방사선조사기술은국제기구 (FAO, IAEA, WHO) 에서유용하고안전한식품및공중보건제품의살균방법으로공인되어이미여러분야에서산업적으로이용되고있다 (7). 최근식품산업에서도방사선조사기술은병원성미생물및유해미생물의사멸에의한위생화, 식량자원의장기보존및손실방지, 국가간식량교역에따른검역관리로써관심이높아지고있으며, 현재 5여개국에서허용하였고 3여개국에서상업적으로실용화되고있다 (8,9). 특히적정선량의감마선조사는식품고유의품질을유지하면서미생물을선택적으로살균할수있고잔류독성이없으며, 제품을완전히포장한후살균이가능하여포장과정에서의 2차오염을방지할수있다 (1). 또한감마선조사는전자선, X-선과비교했을때투과력이매우우수하여과일에조사할경우과피와과육에흡수되는선량이동일하므로과실류의발아억제와해충구제에유리하다 (11). 국제적인교역이활발해지면서각국에서수 출입식품의
감마선조사오렌지의독성평가 1287 검역방법으로방사선조사처리가확대되고있으며, 이에따라검역기준을규정하여이행하고있다 (5). 하지만우리나라는국제교역의증대로식품의수입량이증가함에도불구하고방사선조사식품의유통에대한규정이갖춰져있지않고, 방사선조사식품의안전성에대한부정확한정보로소비자들의태도는여전히부정적이다 (12,13). 또한국외에서는검역을위해과일의방사선조사가허용되고있으며조사량이 이하로규제되고있으나, 국내에서는방사선조사허용식품에과일이포함되어있지않다 (14). 그러므로방사선조사식품에대한다양한연구를수행하여규정을확립하고안전성을입증하여소비자들의수용성을증대시킴으로써이기술이상용화되도록노력해야한다. 방사선조사식품의안전성을입증하고상용화하기위해서는앞서수행한 Song 등 (15) 의감마선조사계란의유전독성학적안전선평가나 Jeon 등 (16) 의감마선조사닭갈비의안전성평가와같은연구가다방면으로이루어져야한다. 따라서본연구에서는국제교역이활발해짐에따라식품에대한방사선조사기술이증대되는데, 이에안전성을검토하고자 감마선조사오렌지의급성및아만성독성평가를수행하였다. 재료및방법시험물질및감마선조사본연구에사용한오렌지는미국캘리포니아산으로시중에서판매중인오렌지를구입하여사용하였다. 감마선조사는한국원자력연구원 (Jeongeup, Korea) 내선원 11.1 PBq, 6 Co 감마선조사시설 (Point Source AECL, IR-79, MDS Nordion International Co., Ltd., Ottawa, Canada) 을이용하였다. 수입오렌지를실온 (14±1 C) 에서시간당 의선량율로 의총흡수선량을얻도록하였다. 흡수선량확인은 alanine dosimeter(5 mm, Bruker Instruments, Rheinstetten, Germany) 를사용하였다. Dosimetry 시스템은국제원자력기구 (IAEA) 의규격에준용하여표준화한후사용하였으며, 총흡수선량의오차는 2% 이내였다. 감마선조사된수입오렌지는껍질을제거한다음동결건조기 (Vacuum Freeze Dryer, Model SFDSF12, Samwon Freezing Co., Seoul, Korea) 에동결건조하여독성시험시료로사용하였다. 실험동물및사육환경실험동물은 4주령의특정병원체부재 (specific pathogen free) 의 ICR 계열마우스를중앙실험동물 ( 주 )(Seoul, Korea) 로부터구입하였다. 7일간의검역및순화과정을거친후체중감소가없는건강한동물을선별하여 5주령때부터시험에사용하였다. 사육환경은온도 23±3 C, 상대습도 6± 1%, 환기횟수및방식 1~2회 / 시간, 전배기방식, 조명시간 12시간 (8:~2:) 및조도 15~3 Lux로설정하 였으며, 모든시험자들은고압증기멸균 (121 C, 2분 ) 한작업복과보호장구를착용하여실험을수행하였다. 실험기간동안동물실의온도와상대습도는자동온습도측정기를이용하여측정하였으며, 환기횟수, 조도등의환경조건은정기적으로측정하였다. 그결과시험에영향을미칠것으로생각되는변화는없었다. 실험동물은폴리카보네이트사육상자 (278 42 2 mm, Three-Shine Inc., Daejeon, Korea) 에사육하였으며, 본실험은한림대학교동물실험윤리위원회의승인 ( 승인번호 : Hallym 214-1) 을받아수행하였다. 시험군구성및시험물질투여동물의군분리는순화기간중건강한것으로판정한동물의체중을측정한후평균체중에가까운동물들을암수각각급성독성은 1마리씩 3개군으로, 아만성독성은 12마리씩 3개군으로나누었다. 급성독성은시험물질을, 1,, 2, mg/kg 용량으로설정하고멸균주사용수에용해하여조제하였으며, 실험동물에게투여하기전하룻밤절식시켜위내용물을비운후경구투여용존데를이용하여위내에직접투여하였다. 투여액량은투여당일에측정한절식시의체중을기준으로하여 1 ml/kg으로하였고, 투여횟수및투여기간은투여당일오전에개체별로단회투여하였다. 아만성독성은 AIN-76 식이를기본으로투여한대조군과감마선비조사 ( kgy) 오렌지투여군, 감마선조사 () 오렌지투여군으로구성하였다. 사료는각각의필수영양소들을 AIN-76 규격에맞게혼합하여 pellet을만들어 3개월간자유로이섭취시켰고, 필터와유수살균기를이용하여여과 살균된정제수를자유급식시켰다. 오렌지의일반성분분석결과탄수화물, 단백질, 지방함량이건물량 (dry basis) 기준으로각각 81.53, 5.17,.34 g/1 g으로확인되었으므로오렌지투여시험군의경우에는오렌지를투여하는대신 AIN-76 식이를기본으로 corn starch, casein 및 soybean oil의양을감소시켜총열량을동일하게조성한 pellet 을만들어사용하였다. 이때오렌지의투여량은사람의섭취량을고려하여 2.5% 로설정하여안전성평가를실시하였다 (Table 1). 사망동물및일반증상관찰급성독성은투여당일에투여후 1시간까지는지속적으로, 투여후 6시간까지는매시간, 투여 1일부터 14일까지는 1회이상일반증상을관찰하였다. 체중은모든동물에대하여투여전, 투여후 1, 3, 7 및 14일에측정하였다. 아만성독성은투여및관찰기간동안사망여부, 일반증상의정도를 1일 1회관찰하고, 개체별로기록하였다. 동물의체중은투여개시일, 그이후에는주 1회및부검일에측정하였다. 부검일은절식시킨체중을측정하였다. 사료및물섭취량은주 1회측정하였다. 측정방법은사료및물을정량급여한후일주일되는날에사육상자단위로측정하여그
1288 정다운 황옥화 최근표 강일준 Table 1. Formula of experimental diets Ingredient Casein AIN-76 mineral mix AIN-76 vitamin mix DL-Methionine Cellulose Corn starch Sucrose Corn oil Choline bitartrate Orange AIN-76 2 35 1 3 5 55 1 5 2 - Orange kgy 198.7 35 1 3 47.5 529.6 1 49.9 2 25 198.7 35 1 3 47.5 529.6 1 49.9 2 25 Total (g) 1, 1,.7 1,.7 차이를계산하고, 마리당일일평균섭취량으로산출하였다. 동물의부검및장기무게사육이끝난실험동물을 12시간동안절식시키고 tribromoethanol(tbe) 로복강마취하였으며, 혈액은안와정맥채혈법으로채취하였다. 급성독성은모든내부장기를대상으로육안적인부검소견을관찰하였고, 아만성독성은간, 비장, 신장, 심장, 폐를적출하여중량을측정하고, 각장기에대하여부검시체중에대한상대중량을산출하였다. 혈액학적검사급성및아만성독성의혈액학적검사는부검시채혈한혈액을 EDTA-2K가들어있는 tube에주입한후자동혈구계측장치 (Hemavet 95, Drew Scientific Inc., Oxford, UK) 를이용하여백혈구 (white blood cell, WBC), 호중구 (neutrophil, NEU), 림프구 (lymphocyte, LYM), 단핵구 (monocyte, MONO), 호산구 (eosinophil, EOS), 호염구 (basophil, BASO) 수치를측정하였다. 나아가적혈구 (red blood cell, RBC), 헤모글로빈 (hemoglobin, Hb), 혈중적혈구비율 (hematocrit, HCT), 평균적혈구용적 (mean corpuscular volume, MCV), 평균적혈구혈색소량 (mean corpuscular hemoglobin, MCH), 평균적혈구혈색소농도 (mean corpuscular hemoglobin concentration, MCHC), 적혈구용적분포폭 (red blood cell distribution width, RDW), 혈소판 (platelet, PLT), 평균혈소판용적 (mean plasma volume, MPV) 수치도측정하였다. 혈청생화학적검사급성및아만성독성의혈청생화학적검사는부검시채혈한혈액을 clot activator가들어있는 tube에주입하고 15~2분간실온에방치하여응고시킨후원심분리하여얻은혈청으로, 혈액생화학분석기기 (KoneLab 2, Thermo Fisher Scientific, Waltham, Finland) 를이용하여 aspartate aminotransferase(ast), alanine aminotransferase (ALT), alkaline phosphatase(alp), total bilirubin(tbil), glucose(glu), total cholesterol(tcho), triglycerides (TG), total protein(tp), albumin(alb), blood urea nitrogen(bun), creatinine(cre) 을측정하였다. 조직학적검사아만성독성의조직학적검사는간, 신장을 4% paraformaldehyde 용액에고정시킨후, Tissue Processor (Leica TP12, Leica, Wetzlar, Germany) 를이용하여파라핀포매를하였다. 포매된조직은 Microtome(Leica RM 2255, Leica, Wetzlar, Germany) 으로 7 μm 박절한후 hematoxylin & eosin 염색을실시하여광학현미경 (Zeiss Axio Imager.M1, Carlzeiss, Oberkochen, Germany) 으로관찰하였다. 통계학적분석이상의실험에서얻어진결과는 Statistical Package for Social Sciences 1.(SPSS Inc., Chicago, IL, USA) 을이용하여 one way ANOVA 분석을하였으며, 시료간의유의성은 Duncan's multiple range test로 P<.5 수준에서비교하였다. 결과및고찰사망동물, 일반증상, 체중변화, 사료섭취량및장기무게급성독성에서 감마선조사오렌지를 ICR 마우스에경구투여한후시험기간동안사망동물은관찰되지않았다 (Table 2). 또한일반증상에서도연변, 설사, 항문주위오염등과관련된어떠한변화도관찰되지않았다 (Table 3). 시험 Table 2. Mortality of ICR mice administered with gamma-irradiated orange for 14 days (mg/kg) 1, 2, 1, 2, Mortality /1 2) /1 /1 /1 /1 /1 1) ALD: approximate lethal dose. 2) Number of dead animals/ Total animals. Days after treatment 1 2 3 4 5 7 8 14 ALD 1) value >2, mg/kg >2, mg/kg
감마선조사오렌지의독성평가 1289 Table 3. Clinical signs in ICR mice administered with gamma-irradiated orange for 14 days Days Signs Normal Soft stool Diarrhea Soiled perineal region (mg/kg) 1, 2, 1, 2, 1/1 1) /1 /1 /1 1 13 Normal 1/1 1/1 1/1 1/1 1/1 1/1 14 Normal Terminal sacrifice 1/1 1/1 1) Number of animals observed/ Number of animals examined. 1/1 /1 /1 /1 1/1 1/1 1/1 /1 /1 /1 1/1 1/1 1/1 /1 /1 /1 1/1 1/1 1/1 /1 /1 /1 1/1 1/1 1/1 /1 /1 /1 1/1 1/1 Table 4. Body weight changes of ICR mice administered with gamma-irradiated orange for 14 days (mg/kg) 1, 2, 1, 2, Days 1 3 7 14 32.96±1.23 NS 35.26±1.75 NS 37.16±2.95 NS 33.18±.87 35.38±1.48 36.5±1.13 33.28±1.13 35.58±1.62 36.5±2.26 28.4±.48 NS2)3) 28.28±.57 28.38±.87 24.26±.92 NS 24.2±1.27 24.3±1.31 28.62±.93 NS 27.5±.96 27.18±1.37 28.58±.88 NS 28.25±1.1 27.68±1.55 1) Weight gains are body weight difference between day 14 and the day. 2) Values are expressed as mean±sd. 3) Not significant at 5% level by Duncan's multiple range test. 29.2±.63 NS 28.8±.34 28.53±1.12 39.6±4.51 NS 36.85±1.8 38.88±3.23 32.26±1.24 NS 31.23±1.73 3.25±2.74 Gains 1) (g) 1.66±4.46 NS 8.58±1.41 1.5±2.38 8.±1.74 NS 7.3±1.15 3.25±2.74 Table 5. Necropsy findings of ICR mice administered with 1 kgy gamma-irradiated orange for 14 days (mg/kg) 1, 2, 1, 2, Fingdings Frequency 1) 1/1 1/1 1/1 1/1 1/1 1/1 1) Number of animals observed/ Number of animals examined. Table 6. Mortality of ICR mice administered with gamma-irradiated orange for 3 months kgy kgy Weeks 1 2 3 1 11 12 13 1) Number of dead animals/ Total animals. Mortality 1) /12 /12 /12 /12 /12 /12 물질투여후 14일간체중변화에서도모두정상적인체중증가가관찰되었으며, 대조군과비교하여통계학적으로유의적인차이를나타내지않았다 (Table 4). 시험기간종료후부검시모든장기에서육안적인이상소견은관찰되지않았다 (Table 5). 따라서사망동물, 일반증상, 체중변화, 부검소견등감마선조사로인한변화로판단될만한요인은발견되지않았다. 아만성독성에서 감마선조사오렌지를 3개월간 ICR 마우스에투여하는동안암수모든시험군에서사망동물은관찰되지않았으며 (Table 6), 육안적인이상소견도관찰되지않았다 (Table 7). 또한암수모든시험군에서정상적인체중증가가관찰되었고감마선조사오렌지투여군을대조군및감마선비조사오렌지투여군과비교해볼때통계학적으로유의적차이가없는것으로나타났으며, 식이섭취량에서도유의적인차이를나타내지않았다 (Table 8). 감 마선조사가주요장기에미치는영향을조사하기위해부검한결과암수모든시험군의장기에서시험물질섭취로인한특이적인이상증상은관찰되지않았으며, 장기무게도통계학적으로유의적인차이를나타내지않았다 (Table 9). 따라서체중변화, 사료섭취량및장기무게모두감마선조사로인한변화로판단될만한요인이발견되지않았다. 혈액학적검사급성독성의혈액학적검사결과암수 ICR 마우스가모든수치에서통계학적으로유의적인차이를보이지않았다 (Table 1). 아만성독성의혈액학적검사결과를 Table 11에나타내었다. 먼저수컷의수치에서 HCT, MCHC 암컷의수치에서 WBC, Hb, MCHC를제외하고는모든수치가통계학적으로유의적인차이를나타내지않았다. HCT, MCHC, WBC, Hb
129 정다운 황옥화 최근표 강일준 Table 7. Clinical signs of ICR mice administered with gamma-irradiated orange for 3 months Weeks Signs kgy kgy 1 Normal 1) 2 12 13 Normal Loss of fur Compound-colored stool Normal Terminal sacrifice /12 /12 1) Number of animals observed/number of animals examined. /12 /12 /12 /12 /12 /12 /12 /12 /12 /12 Table 8. Body weight change and food consumption of ICR mice administered with gamma-irradiated orange for 3 months kgy kgy Initial body weights (g) 37.55±1.66 NS1)2) 37.37±1.76 37.58±.82 29.23±1.27 NS 29.28±1.9 29.92±1.45 1) Values are expressed as mean±sd. 2) Not significant at 5% level by Duncan's multiple range test. Final body weights (g) 56.9±2.26 NS 5.75±2.73 53.33±1.59 44.4±3.4 NS 43.17±.19 45.3±1.54 Body weight gain (g/d).26±.7 NS.17±.5.19±.4.23±.8 NS.2±.5.2±.5 Food consumption (g/d) 4.36±.17 NS 4.49±.19 4.41±.15 3.75±.32 NS 3.94±.22 3.93±.2 Table 9. Relative organ weight of ICR mice administered with gamma-irradiated orange for 3 months Rel. organ wt. (%) 1) kgy kgy Liver Spleen Kidney-left Kidney-right Heart Lung 3.74±1.25 NS2)3).23±.9 NS.61±.15 NS.58±.13 NS.43±.11 NS.44±.1 NS 4.11±.56.28±.7.74±.1.73±.7.47±.7.48±.3 3.74±.45.25±.5.63±.16.59±.14.4±.7.43±.5 3.72±.24 NS.31±.4 NS.45±.9 NS.41±.5 NS.36±.8 NS.48±.14 NS 1) Relative organ weights were expressed as the percentage of organ weights to body weights. 2) Values are expressed as mean±sd. 3) Not significant at 5% level by Duncan's multiple range test. 4.6±.73.41±.17.58±.16.55±.16.44±.9.53±.18 3.9±.62.34±.9.5±.11.47±.11.36±.6.48±.12 Table 1. Hematological test of ICR mice administered with gamma-irradiated orange for 14 days Test WBC NEU LYM MONO EOS BASO RBC Hb HCT MCV MCH MCHC RDW PLT MPV Units M/μL % fl pg % fl (mg/kg) 1, 2, 1, 2, 5.1±.88 NS1)2) 1.59±.53 NS 3.28±.67 NS.12±.8 NS.2±.2 NS.±.1 NS 1.34±.52 NS 14.62±.8 NS 6.82±3.41 NS 58.84±1.63 NS 14.16±.21 NS 24.2±.49 NS 15.38±.64 NS 653.33±16.91 NS 5.4±.13 NS 2.58±1.4.79±.3 1.74±.79.5±.2.1±.1.1±.1 1.37±.7 14.37±.59 57.27±3.18 55.13±1.63 13.18±1.53 23.88±2.54 15.8±.66 648.33±11.29 4.83±.22 1) Values are expressed as mean±sd. 2) Not significant at 5% level by Duncan's multiple range test. 3.71±2.16 1.4±.54 2.54±1.54.8±.7.2±.3.1±.1 1.83±.5 14.57±.97 59.3±4.72 54.8±3.9 13.35±.44 24.33±1.14 15.23±1.14 622.33±139.23 4.9±.1 2.72±.93 NS.43±.17 NS 2.13±.81 NS.12±.4 NS.2±.1 NS.1±.1 NS 9.63±.86 NS 13.8±1.32 NS 59.13±2.65 NS 57.92±4.14 NS 14.32±.41 NS 24.8±1.16 NS 15.28±.41 NS 652.3±17.45 NS 4.96±.22 NS 2.89±1.81.47±.9 2.96±1.97.1±.3.3±.2.3±.4 1.2±.42 14.3±.36 57.68±2.76 56.55±.9 14.3±.39 24.83±.65 15.2±.58 524.25±87.73 4.88±.28 2.17±.99.39±.16 1.69±.86.5±.2.3±.3.1±.3 1.38±.38 14.4±.52 57.65±3.91 55.53±2.17 13.88±.33 25.5±.82 15.88±.88 558.75±125.26 5.5±.1
감마선조사오렌지의독성평가 1291 Table 11. Hematological test of ICR mice administered with gamma-irradiated orange for 3 months Test WBC NEU LYM MONO EOS BASO RBC Hb HCT MCV MCH MCHC RDW PLT MPV Units M/μL % fl pg % fl kgy kgy 4.93±.99 NS1)2).81±.36 NS 2.67±1.1 NS.16±.2 NS.2±.2 NS.1±.1 NS 1.61±.15 NS 12.33±.17 NS 54.43±1.4 b 52.5±3.7 NS 12.88±1.3 NS 22.2±1.25 b 16.7±.57 NS 655.33±119.27 NS 5.23±.35 NS 4.49±1.47 1.3±.67 2.91±1.25.11±.4.2±.1.1±.1 9.74±2.72 13.23±3.39 53.85±3.11 a 51.29±2.55 13.48±.42 26.3±.92 a 16.81±.48 71.8±12.99 4.9±.43 1) Values are expressed as mean±sd. 2) Not significant at 5% level by Duncan's multiple range test. 3.8±1.18 1.7±.48 2.47±.83.9±.6.2±.1.1±.1 1.75±1.1 14.69±1.33 55.3±5.86 a 54.39±2.82 13.58±.31 25.2±1.28 a 16.2±.72 668.75±134.35 4.68±.44 2.38±.33 b.51±.22 NS 1.71±.17 NS.1±.5 NS.5±.3 NS.3±.1 NS 1.3±.52 NS 11.53±1.16 b 5.88±3.7 NS 5.7±1.66 NS 12.73±1.11 NS 22.63±1.3 b 18.18±1.3 NS 571.±74.36 NS 5.8±.29 NS 3.73±1.3 a.63±.32 2.13±.69.1±.8.2±.1.1±.1 1.74±1.36 14.53±1.64 a 51.39±6.98 5.93±4.5 13.26±.9 26.75±1.8 a 17.52±1.2 662.33±12.93 5.±.42 4.±.62 a 1.±.32 2.44±.49 a.1±.5.2±.2.2±.2 11.1±.8 14.9±.72 a 55.48±2.13 52.54±2.96 13.19±.3 25.85±1.26 a 17.41±1.17 674.33±128.54 4.85±.52 수치는 Wolford 등 (17) 의연구에서제시한실험동물의혈액기초자료측정치인 HCT(35.1~55.4%), MCHC(2.2~ 34.2 ), WBC(1.8~1.7 ), Hb(11.~15.1 ) 와비교하였을때모두정상범위내에속하였고, 감마선비조사오렌지군과감마선조사오렌지군은통계학적으로유의적인차이를나타내지않았으므로감마선조사에기인한독성은없는것으로나타났다. 혈액생화학적검사급성독성의혈액생화학적검사결과암수 ICR 마우스가모든수치에서통계학적으로유의적인차이를보이지않았다 (Table 12). 아만성독성의혈액생화학적검사결과를 Table 13에나타내었다. 암수 ICR 마우스의모든수치는통계학적으로유의적인차이를보이지않았다. 다만수컷의 ALT 수치가유의적인차이를나타냈는데이는 Wolford 등 (17) 의연구에서제시한실험동물의혈액기초자료측정치인 ALT(3~ 17 U/L) 와비교하였을때모두정상범위내에속하였고, 감마선비조사오렌지군과감마선조사오렌지군은통계학적으로유의적인차이를나타내지않았으므로감마선조사에기인한독성은아닌것으로나타났다. 따라서 감마선조사오렌지는혈액생화학적측면에서독성이없는것으로판단되었다. 조직학적검사아만성독성에서 감마선조사오렌지를 3개월간섭취한 ICR 마우스의병리학적조직검사를위해우선장기를육안으로관찰한결과모든동물에서장기의형태, 크기, 색조, 경도및기타병변등의육안적이상소견이관찰되지않았다. 이에따라간장및신장을채취하여고정한후염색하여관찰한결과를 Fig. 1과 Fig. 2에나타내었다. 간의경우대조군, 감마선비조사오렌지투여군과감마선조사오렌지투여군모두에서염증, 괴사 bilirubin 침착및 iron 침착등의병적인변화를보이지않았고, 간세포의구조도모 Table 12. Serum biochemistry test of ICR mice administered with gamma-irradiated orange for 14 days Test Units AST U/L ALT U/L ALP U/L TBIL m GLU m TCHO m TG m TP ALB BUN m CRE m (mg/kg) 1, 2, 1, 2, 135.6±51.89 NS1)2) 14.87±24.29 86.13±2.75 131.2±27.3 NS 118.98±27.5 54.93±22.21 NS 7.42±26.45 37.7±.66 36.1±2.38 NS 48.93±24.69 297.6±71.14 NS 322.83±126.64 379.43±91.92 332.21±42.33 NS 393.12±23.78.11±.3 NS.13±.3.11±.2.1±.2 NS.1±.3 18.8±26.99 NS 118.13±41.46 84.8±17.67 112.3±31.83 NS 87.4±13.69 144.46±19.45 NS 122.24±2.75 149.42±14.86 12.97±13.82 NS 82.83±1.89 27.4±3.16 NS 153.59±32.86 25.73±38. 152.5±34.64 NS 164.72±74.84 4.6±.42 NS 4.59±.26 4.76±.9 4.93±.8 NS 4.57±.17 3.35±.32 NS 3.32±.33 3.41±.15 3.57±.23 NS 3.4±.24 22.41±3.98 NS 24.94±5.66 23.41±1.67 16.1±2.59 NS 16.8±4.11.38±.4 NS.37±.5.43±.1.36±.2 NS.36±.2 1) Values are expressed as mean±sd. 2) Not significant at 5% level by Duncan's multiple range test. 131.68±35.37 44.82±9.86 326.76±43.88.11±.4 19.23±14.47 91.78±18.72 179.28±68.26 4.92±.37 3.53±.17 19.5±5.4.36±.3
정다운 황옥화 최근표 강일준 1292 Table 13. Serum biochemistry Test Units (mg/kg) U/L AST ALT U/L ALP U/L TBIL m GLU m TCHO m TG m TP ALB BUN m CRE m 1) 2) test of ICR mice administered with gamma-irradiated orange for 3 months kgy 1, 2, 116.72±51.1NS1)2) 172.7±49.62 171.17±32.24 145.77±32.1NS 151.28±31.12 155.89±3.6 75.11±21.59a 77.47±14.84a 48.8±15.21NS 5.±15.14 5.63±18.71 42.38±7.36b NS 164.3±27.47 159.54±25.6 159.76±28.42 21.16±21.2NS 174.18±36.58 173.49±27.86.14±.1NS.16±.4.17±.5.1±.2NS.11±.2.11±.2 NS 145.85±27.2 122.61±27.77 121.43±15.4 111.79±11.62NS 16.4±18.9 18.4±16.6 168.17±72.94NS 167.94±34.7 175.11±31.67 95.15±2.2NS 92.73±34.24 96.87±18.87 77.8±25.33 77.61±27.67 122.15±15.44NS 11.72±27.14 111.±17.87 77.66±1.65NS 5.38±.36NS 5.2±.19 5.12±.41 5.15±.61NS 5.1±.53 5.±.42 3.47±.34NS 3.21±.27 3.38±.34 3.68±.25NS 3.41±.38 3.39±.23 19.34±2.59 22.68±2.22NS 22.84±5.15 22.4±3.59 16.79±2.49NS 2.5±5..45±.NS.41±.7.44±.5.45±.4NS.42±.4.43±.5 Values are expressed as mean±sd. Not significant at 5% level by Duncan's multiple range test. 두 정상이었다(Fig. 1). 또한 신장에서도 염증 및 괴사 등의 에 기인하는 괴사도 관찰되지 않았다(Fig. 2). 따라서 병적인 변화를 보이지 않았고, 면역학적 요인이나 독성물질 감마선 조사 오렌지를 3개월간 암수 ICR 마우스에게 섭취시 kgy Fig. 1. Histopathological examination of liver in the ICR mice administered with gamma-irradiated orange for 3 months, 2. kgy kgy kgy Fig. 2. Histopathological examination of kidney in the ICR mice administered with gamma-irradiated orange for 3 months, 2.
감마선조사오렌지의독성평가 1293 켜도조직학적측면에서는독성이나타나지않는것으로사료된다. 이상의독성시험결과 감마선조사오렌지를 ICR 마우스에투여했을때어떤유의할만한변화를나타내지않았음을관찰할수있었다. 즉 감마선조사오렌지를이용하여암수 ICR 마우스에급성, 아만성독성평가를수행한결과독성이없는것으로판명되었다. 이는이온화에너지를조사한수입오렌지의유전독성학적안전성평가에서이온화에너지조사수입오렌지가돌연변이원성이없고염색체이상을나타내지않았으며, 소핵을유발하지않아유전독성학적측면에서안전성이확인된 Huang 등 (18) 의연구와도잘일치하였다. 또한오렌지를 2.97 kgy로감마선조사한후 rat에게 16일간섭취시켜번식시험과최기형성시험을수행한결과체중이약간감소한것을제외하고는감마선조사의독성이관찰되지않았다 (19). 그밖에감마선조사타락죽의독성평가를수행한 Yin 등 (2) 과감마선조사곡류분말의독성평가를수행한 Jeon 등 (21) 의연구에서도감마선조사로인한비조사군과조사군사이의변화가나타나지않아독성이없다고보고하였으며, 이는본시험의결과와유사한결과를나타내었다. 요약본연구에서는방사선조사식품의규정을확립하고안전성을입증하기위해 감마선조사오렌지를 ICR 마우스에투여또는섭취시켜급성및아만성독성시험을실시하였다. 급성독성의경우시험물질을, 1,, 2, mg/kg의용량으로설정하여단회투여한후 14일간사망동물, 일반증상, 체중변화, 부검소견, 혈액학적및혈액생화학적검사를수행한결과어떠한이상도관찰되지않았으며, 통계학적으로도유의적인차이를나타내지않았다. 따라서 감마선조사오렌지의대략의치사량 (approximate lethal dose) 은암수 ICR 마우스모두 2, mg/kg을상회하는것으로판명되었다. 아만성독성은시험물질을식이에함유하도록하여 3개월간섭취시킨후사망동물, 일반증상, 체중변화, 사료섭취량, 장기무게, 조직학적검사, 혈액학적및혈액생화학적검사를수행하였다. 시험기간동안암수 ICR 마우스모두에서사망동물이나이상증상은발견되지않았으며, 체중변화, 사료섭취량및장기무게에서도통계학적으로유의적인차이를보이지않았다. 혈액학적및혈액생화학적검사에서도암수 ICR 마우스모두정상적인수치를나타냈다. 또한조직학적검사에서간, 신장조직은모두정상적인구조를유지하고있었으며, 염증, 괴사등의유의할만한병적변화도관찰되지않았다. 따라서 감마선조사오렌지를 ICR 마우스에 3개월간섭취시켜도본시험조건에서는독성이없는것으로판명되었다. 감사의글 본연구는농림수산식품부수출전략기술개발사업의위탁과제에의하여수행되었으며지원에감사드립니다. REFERENCES 1. Pala CU, Toklucu AK. 213. Microbial, physicochemical and sensory properties of UV-C processed orange juice and its microbial stability during refrigerated storage. LWT- Food Sci Technol 5: 426-431. 2. Moon H, Lee HK, Park H. 213. Impacts of the KORUS FTA's orange import tariff-cut on domestic fruit prices. Korean J Agricultural Economics 54: 15-38. 3. Kang HJ, Chung HS, Jo DJ, Byun MW, Choi SJ, Choi JU, Kwon JK. 23. Effects of gamma radiation and methyl bromide fumigation on physiological and chemical quality of apples. Korean J Food Preserv 1: 381-387. 4. UNEP. 1994. Montreal protocol on substances that deplete the ozone layer. Report of the Methyl Bromide Technical Options Committee, Nairobi, Kenya. p 294. 5. Kyung EJ, Kim KH, Yook HS. 214. Quality characteristics of gamma irradiated-imported orange during storage at room temperature (2 C). Korean J Food & Nutr 27: 183-193. 6. Ko J, Ma Y, Song KB. 25. Effect of electron beam irradiation on the microbial safety and qualitied of sliced dried squid. J Korean Soc Food Sci Nutr 34: 433-437. 7. Byun MW, Yook HS. 23. Internal and external situation of irradiation technology utilization in the food and public health industry. Korean J Food Preserv 1: 16-123. 8. Byun MW, Yook HS, Jo SK, Chong YJ. 1996. Status and prospects of food irradiation technology in Korea. J Food Sci Nutr 1: 262-268. 9. Kume T, Furuta M, Todoriki S, Uenoyama N, Kobayashi Y. 29. Status of food irradiation in the world. Radiat Phys Chem 78: 222-226. 1. Byun MW. 1997. Application and aspect of irradiation technology in food industry. Food Science and Industry 3(1): 89-1. 11. Jo DJ, Kown JH. 23. Detection of radiation induced markers in oranges imported from united states of America. J Korean Soc Food Sci Nutr 32: 1-7. 12. Lee JW. 29. International cooperation for establishing SOP on quarantine management of irradiated food in international trade. Korea Atomic Energy Research Institute, Daejeon, Korea. p 1-3. 13. Nam HS, Kim KE, Yang JS, Ly SY. 2. Food majoring college students' knowledge and acceptance of irradiated food. Korean J Dietary Culture 15: 269-277. 14. Delincee H. 1998. Detection of irradiated food: DNA fragmentation in grapefruits. Radiat Phys Chem 52: 135-139. 15. Song HP, Shin EH, Yun HJ, Jo C, Kim D. 29. Establishing the genotoxicological safety of gamma-irradiated egg white and yolk. Korean J Food Preserv 16: 782-788. 16. Jeon YE, Yin XF, Kim TK, Kang IJ. 213. Safety evaluation of 3 kgy-irradiated Dakgalbi. J Korean Soc Food Sci Nutr 42: 1475-1481. 17. Wolford ST, Schroer RA, Gohs FX, Gallo PP, Brodeck M, Falk HB, Ruhren R. 1986. Reference range data base for serum chemistry and hematology values in laboratory animals. J Toxicol Environ Health 18: 161-188. 18. Huang YH, Jung DW, Kang IJ. 214. Genotoxicological
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