J Koren So Food Si Nutr 한국식품영양과학회지 38(5), 601~605(2009) DOI: 10.3746/jkfn.2009.38.5.601 이산화염소수처리에의한잔류농약분해효과 김규리 송경빈 충남대학교식품공학과 Effet of Aqueous Chlorine Dioxide Tretment on the Deomposition of Pestiide Residues Kyuri Kim nd Kyung Bin Song Dept. of Food Siene nd Tehnology, Chungnm Ntionl University, Dejeon 305-764, Kore Astrt This study ws onduted to exmine the effet of queous hlorine dioxide tretment s wshing method on removl of pestiide residues. Three pestiides of hlorpyrifos, dizinon, nd metlxyl, whih re ommonly used in vegetle rops, were treted with 10, 50, nd 100 ppm of queous hlorine dioxide nd deomposition of the pestiides ws determined using gs hromtogrphy. Three pestiides used in this study were deomposed y queous hlorine dioxide tretment nd removl rte ws proportionl to tretment time s well s onentrtion of queous hlorine dioxide. In prtiulr, 100 ppm of queous hlorine dioxide tretment deresed the pestiides effiiently. In ddition, lettue ws treted y dipping in distilled wter nd 100 ppm queous hlorine dioxide, respetively, nd ws ompred regrding removl effiieny of the pestiides. The results reveled tht wshing with 100 ppm queous hlorine dioxide for 10 min ws the most effetive for removing the pestiides. These results suggest tht queous hlorine dioxide n e used s wshing method of fresh produe to remove the residul of pestiides. Key words: queous hlorine dioxide, pestiides, wshing tretment 서론농산물생산에있어서농약의사용은필수불가결하지만최근농약의남용에의한저항성병해충의발생이증가하고, 또한토양이나수질의오염, 잔류농약에의한식품의안전성을위협하는사례가발생하고있다 (1). 따라서안전한농산물을공급하고자, 농산물에서의잔류농약피해를최대한줄이기위한농약사용에대한기준을설정하고대상작물, 사용시기, 사용량, 사용횟수등을정하고있으며식품에대한잔류허용기준도설정하고있다 (2). 농산물에남아있는잔류농약은수확후수세, 다듬기, 데치기, 가열등가공공정중많은양이제거되는것으로알려져있으나 (3,4), 이러한공정을거치지않는과일이나채소류같은경우수세나세제를사용하여농약을제거해야하는데, 오존수처리같은화학적처리에의한농약제거와관련된연구가많이진행되어왔다 (5-7). 농약제거와관련한산화제등화학적처리방법과관련하여, 이산화염소수를생각할수있는데, 이산화염소수는식품가공공정에서이미사용되고있는식품표면살균제로강한산화력을가지고있고또한염소와는달리 trihlomethnes 와같은유해물질생성량이없다는장점도있다 (8-10). 특히 FDA가과일과채소의세척에이산화염소수를허용한이후식품산업에서의실용화가증가하고있는데 (11), 이산화염소수에의한잔류농약제거와관련된연구는아직보고된바가없다. 따라서본연구에서는채소에서검출빈도가높은유기인계농약으로 hlorpyrifos, dizinon과페닐아마이드계농약인 metlxyl(fig. 1) 을대상으로이산화염소수처리에의한농약분해효과를측정하였고또한상추를대상으로잔류농약제거효과를조사하였다. 재료및방법실험재료본실험에사용된상추는대전지역에서구입하여농약이검출되지않은것을사용하였다. 농약분해관련하여사용된농약은더스반 (Dursn, hlorpyrifos 25%, Dongu Hnnong Chem., Seoul, Kore), 다이아톤 (Ditone, dizinon 34%, Sungo Chem., Goyng, Kore) 과리도밀 (Ridomyl, metlxy 25%, Dongu Fine Chem., Seoul, Kore) 을사용하였 Corresponding uthor. E-mil: ksong@nu..kr Phone: 82-42-821-6723, Fx: 82-42-825-2664
602 김규리 송경빈 () () () Fig. 1. Chemil struture of pestiides. () hlorpyrifos () dizinon () metlxyl. 다. 표준검량선을작성하기위한농약의표준품은 Dr. Ehrenstorfer GmH(Augsurg, Germny) 의제품을사용하였고, etonitrile, etone, n-hexne은 HPLC용 (J.T. Bker, Phillipsurg, NJ, USA) 을사용하였고, SPE-Florisil 은 Phenomenex(8B-S013-JCH, Torrne, CA, USA) 제품을사용하였다. 이산화염소수의제조이산화염소수처리를위한이산화염소수는 Chlorine dioxide genertor system(ch 2O In., Olympi, Wshington, USA) 을이용하여제조하였으며 (12), 이산화염소수농도는 iodometry stndrd method(13) 를사용하여제조하였다. 표준검량선의작성본연구에사용된 hlorpyrifos, dizinon, metlxyl의표준검량선을작성하기위해각성분의표준용액을 hlorpyrifos, dizinon의경우 0.005,, 0.2,, 1 μg/ml의농도로 metlxyl의경우, 0.2,, 1, 2 μg/ml의농도로희석하여 1 μg을 GC/NPD(Agilent 6890, Snt Clr, Cliforni, USA) 에주입하여각성분 pek의 height에의하여표준검량선을작성하였다. Chlorpyrifos, dizinon, metlxyl의표준검량선작성을위해 X축을표준용액의농도, Y축을 pek 의 height 값으로하여작성한결과, 3 종의농약모두상관계수가 0.999 이상을나타내었는데, GC/NPD 및농약성분의확인을위한 GC/MSD(Vrin CP-3800, Cliforni, USA) 분석조건은 Tle 1과같다. 이산화염소수의농약분해능측정 Nlgene ottle에 distilled wter와 10, 50, 100 ppm의이산화염소용액 200 ml에 100 ppm으로희석된 hlorpyrifos, dizinon, metllxyl 용액 200 μl를첨가하여농약의최종농도가 ppm이되도록조제한후일정한시간 (0, 3, 5, 7, 10, 20, 30 min) 간격으로 25 ml씩취하여농약성분분석을 GC-NPD로분석하였다. 상추에서의잔류농약분석상추를각각사용기준농도에따라제조된 hlorpyrifos, dizinon, metlxyl 용액을담은수조에서 1분동안잘저어주면서침지시킨후꺼내어건조시켰다. 그리고건조시킨상추 50 g을 distilled wter와농도별이산화염소수용액 4 L에 5분, 10분동안각각침지하여세척하고분쇄한다음상추 25 g을취해 etonitrile 100 ml에넣고균질기 (Omni Interntionl, Wterury, CT, USA) 로 3분간균질화하고여과한액을 1분동안진탕한후 4 o C, 6,000 rpm으로 3분간원심분리하고 etonitrile층에서 10 ml를취해서농축한후 3.5 ml 20% etone/n-hexne에용해하고, FLO florisil rtridge에 loding하여수집된 7 ml 용액을다시질소가스를이용하여미세농축한후 etone 2.5 ml에다시녹인시료를 GC-NPD로분석하였다. Tle 1. GC/NPD nd GC/MSD opertion ondition GC/NPD GC/MSD Instrument Agilent 6890N VARIAN CP-3800/Sturn 2200 DB-17MS pillry olumn VF-5MS pillry olumn Column (30 m 0.2 mm (i.d) 0.25 μm) (30 m 0.2 mm (i.d) 0.25 μm) Inlet temperture 250 o C Column flow 1.0 ml/min Injetion volume 1 μl Split rtio 3:1 5:1 Crrier gs N 2 (1.0 ml/min) He (1 ml/min) Detetor Oven Temperture: 325 o C H 2 flow: 3.0 ml/min Air flow: 60 ml/min SIM mode Chlorpyrifos (m/z): 97, 199, 258, 314 Dizinon (m/z): 137, 153, 179, 304 Metlxyl (m/z): 45, 160, 192, 206 150 o C (2 min hold) 15 o C/min 200 o C (0 min hold) 10 o C/min 280 o C (10 min hold) Post run: 300 o C (2 min hold)
이산화염소수처리에의한잔류농약분해효과 603 Tle 2. Reovery, LOD, nd LOQ of the nlytil method Pestiides Fortifition (mg/kg) Reovery±RSD (%) 1) LOD 2) (mg/kg) LOQ 3) (mg/kg) Chlorpyrifos 98.5±2.57 94.9±4.00 0.005 0.01 Dizinon 104.9±2.87 95.0±2.63 0.005 0.01 Metlxyl 98.7±6.31 95.3±1.53 0.3 1) Men vlues±stndrd devitions. Limits of detetion. Limits of quntifition. 회수율, 검출한계, 정량한계측정본실험에서사용된 hlorpyrifos, dizinon, metlxyl의잔류농약분석법에대한회수율을측정하기위해농약이검출되지않은상추에 3가지농약을, mg/kg씩을각각첨가한후시료의분석방법과동일하게 3회반복하여실험하여 Tle 2에나타내었다. 검출한계 (limits of detetion, LOD) 와정량한계 (limits of quntifition, LOQ) 를측정하기위해각농약성분의한계희석액을 5회반복하여얻은값으로다음과같은식을이용하여구하였다. LOD=3.3 ( 표준편차 / 검량선의기울기 ) LOQ=10 ( 표준편차 / 검량선의기울기 ) 통계분석실험결과의유의성검정은 SAS progrm(14) 을사용하여 p<0.05 수준에서 Dunn s multiple rnge test 방법을사용하여통계처리를하였다. 실험결과는평균 ± 표준편차로나타냈다. 결과및고찰이산화염소수의농약분해능측정이산화염소수에의한농약분해능을측정한결과, 세가지농약성분모두처리시간이증가함에따라빠른속도로분해되기시작하였고또한이산화염소수농도에비례하여 100, 50, 10 ppm 순으로분해속도가빨랐다 (Fig. 2). Chlorpyrifos 의경우초기농도인 5 ppm에서처리시간이 5분일때 100 ppm 이산화염소수처리에서의농도가 0.02 ppm으로줄어 72% 의농약분해율을보였고처리시간이 20분경과시검출한계미만으로나타났다. Dizinon 의경우, 초기농도인 2 ppm에서처리시간이 5분에서는 100 ppm 이산화염소수처리경우, 농약농도가 0.01 ppm이조금넘어 86% 의분해율을보였고처리 7분후에는거의모두분해되어검출한계미만으로나타남을확인할수있었다. 그러나 metlxyl의경우에는초기농도인 5 ppm에서처리시간이 10분시 100 ppm 이산화염소수에서의농약농도가 0 ppm이되어 33% 의분해율을보여가장낮은분해율을보였다. 이러한실험결과에서이산화염소수처리가농약성분을분해하는데효과가있음을확인할수있었고, 특히처리이산화염소수농도가높을수록농약분해율도높은것을알수있었다. 또한 hlorpyrifos와 dizinon의경우에는이산화 염소수처리농도 50 ppm과 100 ppm에서는농약분해에있어서큰차이를보이지않았으며, 3가지농약중 metlxyl의경우가가장낮은분해율을보여이산화염소수처리에있어서유기인계농약이페닐아마이드계농약보다분해율측면에서보다바람직하다고판단된다. Kim 등 (15) 의농약에오염된콩나물에오존수를처리한연구결과에서, 농약에있어서그화학조성이 P=O 결합, S=P 결합, C=O 결합순으로농약분해율이감소한다고했는데, 본실험에사용된농약의경우 dizinon, hlorpyrifos는 S=P 결합을가지고있고 metlxyl은 C=O결합을가지고있어, 본연구에서수행된이산화염소수처리에의해분해되는대상농약의분해율결과와일치함을보여준다. 따라서강산화제인이산화염소수처리효과도오존수와유사한산화기작으로농약을분해하는것으로판단되는데, 오존은강력한산화제로써농약의이중결합에작용하여산화개열반응과방향족화합물의개환및측쇄부위의산화등다양한반응을일으키는것으로알려져있다 (16-18). 상추에서의세척방법에따른잔류농약제거농약이검출되지않은신선한상추에인위적으로농약 3 종을오염시킨후세척방법에따른농약분해율을측정한결과 (Fig. 3), 이산화염소수 100 ppm으로 10분처리한경우가모든농약에대해농약분해정도가유의적으로가장큰것을알수있었는데이것은 Fig. 2에서의결과와도동일한패턴을보이는것이다. Chlorpyrifos의경우 100 ppm 이산화염소수 10분처리시 87% 의제거율을보였고 dizinon의경우는 73% 의제거율을보였으며 metlxyl의경우는 79% 의제거율을보였다. 세척방법별상추에잔류하는농약의잔류정도는농약별로차이가있었지만, 모든농약에대해물세척에비하여이산화염소수를사용한세척방법이농약성분제거에유의적으로효과가있음을알수있었다. 이러한결과는과일과채소표면의잔류농약제거에물세척만으로는효과가적다는보고 (19) 를감안할때, 신선한채소와과일의섭취에있어서잔류농약의위험성을낮추기위해서는이산화염소수세척처리가식품의안전성측면에서바람직하다는것을시사한다. 또한, 농약성분별세척율에있어서 3가지농약중물에대한용해성이가장적은 hlorpyrifos가가장높아농약의수용성이세척효과와비례하지않음을알수있었는데이것은세척에의한농약의제거효율은약제의수용성정도
604 김규리 송경빈 () 0 250 Pestiide onentrtion (ppm) 0.06 0.04 0.02 Pestiide onentrtion (ppm) 200 150 100 50 d d e () Pestiide onentrtion (ppm) () Pestiide onentrtion (ppm) 0.00 6 4 2 0 0.06 0.04 0.02 0.00 6 4 2 0 Fig. 2. Deomposition of pestiides y queous hlorine dioxide tretment. Brs represent stndrd error. () hlorpyrifos () dizinon () metlxyl. : distilled wter : 10 ppm : 50 ppm : 100 ppm. 보다는침투성등농산물의특성에의해영향을받는다는연구와일치하였다 (20-22). 그리고같은세척방법이라고해도세척시간이 5분에서 10분으로처리시간이길수록상추에서의농약제거율이더높았는데 (Fig. 3), 이것은깻잎에서의잔류농약이세척시간증가에따라농약제거율이증가한다는연구와유사하였다 (23). 결론적으로본연구결과, 상추등농산물에서의잔류농약제거를위해서는단순한물세척보다는이산화염소수 100 0 Dizinon Metlxyl Chlorpyrifos Fig. 3. Deomposition of pestiide residues in lettue y vrious wshing tretments. : Control, : Wter (5 min), : Wter (10 min), : ClO 2 100 ppm (5 min), : ClO 2 100 ppm (10 min). ppm으로세척하는것이식품안전성측면에서더바람직하다는것을보여준다. 요 본연구는이산화염소수처리가식품에서의잔류농약제거에효과가있는지조사하기위해서수행되었다. 10, 50, 100 ppm 이산화염소수로채소류에흔히쓰이는농약인 hlrpyrifos, dizinon, metlxyl을처리하여 GC를이용하여농약의분해율을측정하였다. 상기세가지농약성분은이산화염소수처리농도뿐만아니라처리시간에비례하여분해되었으며특히 100 ppm 이산화염소수처리가가장효과적이었다. 또한, 인위적으로농약을오염시킨상추를증류수, 100 ppm 이산화염소수에각각침지하여농약의제거율을비교해보았을때, 100 ppm 이산화염소수에 10분동안침지하였을때가가장효과가좋았다. 본연구결과, 이산화염소수처리는신선농산물에서의잔류농약을제거하기위한세척수로이용될수있을것으로판단된다. 문 1. Rurl Development Administrtion. 1999. Integrted Pest Mngement: Bsi onepts nd prties. Rurl Development Administrtion, Kore. p 153-179. 2. Lee JM, Lee HR, Nm SM. 2003. Removl rte of residul pestiides in perill leves with vrious wshing methods. Koren J Food Si Tehnol 35: 586-590. 3. Choi KI, Seong KY, Jeong TG, Lee JH, Hur JH, Ko KY, Lee KS. 2002. Dissiption nd removl rte of dihloflunid nd iprodione residues on greenhouse herry tomto. Koren J Environ Agri 21: 231-236. 4. Nm SM, Lee HR, Lee JM. 2003. Removl effiieny of residul pestiides during proessing of Perill Jnghi preprtion. Koren J Food Culture 18: 562-568. 5. Li PK, Li QZ, Liu F, M LN, Li HW, Sho YL. 2004. Study 약 헌
이산화염소수처리에의한잔류농약분해효과 605 on lening out residue of orgnophosphorous insetiides in peh. Pestiide Si Admin 25: 16-20. 6. Yu X-Y, Chen F, Xu D-M, Liu X-J, Zhng X. 2005. Removl of 3 orgnophosphorous insetiide residues with ozone nd its influene on the ontent of V nd rotenoid in vegetle. J Northwest Si Teh Univ Agri Forestry 33: 150-154. 7. Zhng CZ, Luo AL, Wng DL, Liu XJ. 2004. Degrdtion method of et-ypermethrin residue on green groery. J Agro-Environ Si 24: 196-200. 8. Wu VCH, Kim B. 2007. Effet of simple hlorine dioxide method for ontrolling five foodorne pthogens, yests nd molds on lueerries. Food Miroiol 24: 794-800. 9. Chun H, Song KB. 2007. Aqueous hlorine dioxide tretment improves the shelf life of Pnx ginseng C.A. Meyer. J Food Si Nutr 12: 284-288. 10. Hong Y, Ku K, Kim M, Won M, Chung K, Song KB. 2008. Survivl of Eshehi oli O157:H7 nd Slmonell typhimurium, inoulted on hiken y queous hlorine dioxide tretment. J Miroiol Biotehnol 18: 742-745. 11. Food nd Drug Administrtion. 2007. Guidne for industry-guide to minimize miroil food sfety hzrds for fresh fruits nd vegetles. Aville from: http://www.fsn.fd.gov Aessed Fed. 27 12. Youm HJ, Ko JK, Kim MR, Song KB. 2004. Inhiitory effet of queous hlorine dioxide on survivl of Eshehi oli O157:H7, Slmonell typhimurium, nd Listeri monoytogenes in pure ell ulture. Kore J Food Si Tehnol 36: 514-517. 13. APHA. 1995. Stndrd methods for the exmintion of wter nd wstewter. 19th ed. Amerin Puli Helth Assoition. Method 4-54. Wshinhton DC, USA. 14. SAS Institute. 2001. SAS system for windows. Version 8.2. SAS Institute, In., Cry, NC, USA. 15. Kim SD, Kim ID, Prk MZ, Lee YG. 2000. Effet of ozone wter on pestiide-residul ontents of soyen sprouts during ultivtion. Koren J Food Si Tehnol 32: 277-283. 16. Lplnehe GM, Tonnrd F. 1984. Ozontion shemes of orgnophosphorous pestiides-pplition in drinking wter tretment. Ozone Si Eng 6: 207-219. 17. Rie RG, Browning ME. 1981. Ozone tretment of industril wste wter. Noyes Dt orportion, Prk Ridge, NJ, USA. p 85-99. 18. Benoit-Guyod JL, Crosy DG, Browers JB. 1986. Degrdtion of MCP y ozone nd light. Wter Res 20: 67-72. 19. Krieger RI, Brutshe-Keiper P, Crosy HR, Krieger AD. 2003. Redution of pestiide residues of fruit using wter only or plus fit fruit nd vegetle wsh. Bull Environ Contm Toxiol 70: 213-218. 20. Crs P, Angioni A. 2000. Pestiide residues in grpes, wine, nd their proessing produts. J Agri Food Chem 48: 967-973. 21. Krol WJ, Arsenult TL. 2000. Redution of pestiide residues on produe y rinsing. J Agri Food Chem 48: 4666-4670. 22. Lee HD, You OJ, Ihm YB, Kwon HY, Jin YD, Kim JB, Kim YH, Prk SS, Oh KS, Ko SL, Kim TH. 2006. Residul hrteristis of some pestiides in/on pepper fruits nd leves y different types, growing nd proessing onditions. Koren J Environ Agri 10: 99-106. 23. Seo JM, Kim JP, Yng YS, Oh MS, Chung JK, Shin HW, Kim SJ, Kim ES. 2007. The degrdtion ptterns of three pestiides in perill lef y ultivtion, storge nd wshing. J Food Hygiene Sfety 22: 199-208. (2009 년 4 월 9 일접수 ; 2009 년 4 월 27 일채택 )