commiss direct ec adapt technic progress time council direct eec approxim law regul administr provis relat classif packag label danger substanc text eea relev commiss european commun regard treati establish european commun regard council direct eec june approxim law regul administr provis relat classif packag label danger substanc amend european parliament council direct ec articl thereof annex direct eec list danger substanc particular classif label substanc present scientif technic knowledg shown list danger substanc annex adapt languag version direct requir correct specif section foreword tabl annex annex iii direct eec list phrase indic natur special risk attribut danger substanc prepar annex iv direct eec list phrase indic safeti advic danger substanc prepar annex vi direct eec guid classif label danger substanc prepar languag version direct requir correct specif section annex iii iv vi annex direct eec lai method determin physico chemic properti toxic ecotox substanc prepar adapt annex technic progress annex ix direct eec provis relat child proof fasten provis adapt updat extend scope child proof fasten measur provid direct accord opinion committe adapt technic progress direct elimin technic barrier trade danger substanc prepar adopt direct articl direct eec amend annex amend note annex direct replac note foreword row annex direct replac row tabl entri annex direct replac entri entri annex direct insert risk phrase annex direct replac phrase annex iii annex iv amend safeti phrase annex direct replac phrase annex iv combin safeti phrase annex direct replac phrase annex iv part annex amend text annex direct replac chapter text annex direct replac chapter text annex direct replac chapter text annex direct replac chapter text annex direct replac chapter text annex direct replac chapter titl chapter explanatori note chang text annex direct ad fourth indent gener introduct part annex delet text annex direct replac text annex vi annex ix amend set annex direct articl member state bring forc law regul administr provis compli direct june latest forthwith inform commiss thereof member state adopt provis refer direct accompani refer occas offici public member state determin refer made member state commun commiss main provis nation law adopt field cover direct correl tabl direct nation provis adopt articl direct enter forc dai public offici journal european commun articl direct address member state brussel commiss margot wallström member commiss adopt adapt oj oj annex foreword annex explan note relat identif classif label substanc da note klassificeringen som kræftfremkaldend kan udelad fibr som opfyld en af følgend betingels en kortvarig biopersistensprøv ved inhal har vist fibr der er længere end µm har en vægtet halveringstid på mindr end dage en kortvarig biopersistensprøv ved intratrak instil har vist fibr der er længere end µm har en vægtet halveringstid på mindr end dage en egnet intra periton prøve ikk har vist kræftfremkaldend virkn eller en egnet langvarig inhalationsprøv ikk har vist relevant sygdomsfremkaldend virkning eller neoplastisk forandring sv note ämnet behöver int klassificera som cancerframkalland om det kan visa att det uppfyl ett av följand villkor ett korttidstest för att bestämma den biologiska beständigheten vid inhal har visat att fibrer längre än µm har en viktad halveringstid på mindr än dagar ett korttidstest för att bestämma den biologiska beständigheten vid intratrak instil har visat att fibrer längre än µm har en viktad halveringstid på mindr än dagar ett lämpligt intraperitonealt test har int givit belägg för förhöjd cancerogenitet frånvaro av relev patogenitet eller neoplastiska förändringar ett lämpligt långtid inhalationstest concern es version concern de version concern el version concern en version concern fr version concern version concern nl version concern pt version concern fi version annex quot tabl gt tabl gt quot annex gt tabl gt annex gt tabl gt annex gt tabl gt concern es version concern da version concern de version concern el version concern en version concern fr version concern nl version concern pt version concern fi version concern sv version annex gt tabl gt concern es version concern da version concern de version concern el version concern en version concern version concern nl version concern pt version concern fi version concern sv version gt tabl gt concern es version concern da version concern el version concern en version concern fr version concern version concern nl version concern pt version concern fi version concern sv version gt tabl gt concern es version concern da version concern el version concern fr version concern nl version concern pt version concern fi version concern sv version annex gt tabl gt concern es version concern da version concern el version concern en version concern fr version concern version concern nl version concern pt version concern fi version concern sv version gt tabl gt concern da version concern el version concern fr version concern pt version concern fi version annex quot mutagen vitro mammalian chromosom aberr test method method replic oecd tg vitro mammalian chromosom aberr test introduct purpos vitro chromosom aberr test identifi agent structur chromosom aberr cultur mammalian cell structur aberr type chromom chromatid major chemic mutagen induc aberr chromatid type chromosom type aberr occur increas polyploidi chemic potenti induc numer aberr method design measur numer aberr routin purpos chromosom mutat relat event human genet diseas substanti evid chromosom mutat relat event caus alter oncogen tumour suppressor gene somat cell involv cancer induct human experiment anim vitro chromosom aberr test emploi cultur establish cell line cell strain primari cell cultur cell select basi growth abil cultur stabil karyotyp chromosom number chromosom divers spontan frequenc chromosom aberr test conduct vitro gener requir exogen sourc metabol activ metabol activ system mimic mammalian vivo condit care avoid condit lead posit result reflect intrins mutagen aris ph osmol high level cytotox test screen mammalian mutagen carcinogen compound posit test mammalian carcinogen perfect correl test carcinogen correl depend chemic class increas evid carcinogen detect test act mechan direct dna damag gener introduct part definit chromatid type aberr structur chromosom damag express breakag singl chromatid breakag reunion chromatid chromatid type aberr structur chromosom damag express breakag breakag reunion chromatid ident site endoredupl process period dna replic nucleu mitosi start period result chromosom chromatid gap achromat lesion smaller width chromatid minimum misalign cromatid mitot index ratio cell metaphas divid total number cell observ popul cell indic degre prolifer popul numer aberr chang numer chromosom normal number characterist cell utilis polyploidi multipl haploid chromosom number diploid number structur aberr chang chromosom structur detect microscop examin metaphas stage cell divis observ delet fragment intrachang interchang principl test method cell cultur expos test substanc metabol activ predetermin interv exposur cell cultur test substanc treat metaphas arrest substanc colcemid colchicin harvest stain metaphas cell analys microscop presenc chromosom aberr descript test method prepar cell varieti cell line strain primari cell cultur includ human cell chines hamster fibroblast human mammalian peripher blood lymphocyt media cultur condit cultur media incub condit cultur vessel concentr temperatur humid maintain cultur establish cell line strain check routin stabil modal chromosom number absenc mycoplasma contamin contamin normal cell cycl time cell cultur condit prepar cultur establish cell line strain cell propag stock cultur seed cultur medium densiti cultur reach confluenc time harvest incub lymphocyt blood treat anti coagul heparin separ lymphocyt obtain healthi subject ad cultur medium mitogen phytohaemagglutinin incub metabol activ cell expos test substanc presenc absenc metabol activ system commonli system cofactor supplement post mitochondri fraction prepar liver rodent treat enzym induc agent aroclor mixtur phenobarbiton naphthoflavon post mitochondri fraction concentr rang final test medium condit metabol activ system depend class chemic test case utilis concentr post mitochondri fraction number develop includ construct genet engin cell line express specif activ enzym provid potenti endogen activ choic cell line scientif justifi relev cytochrom isoenzym metabol test substanc test substanc prepar solid test substanc dissolv suspend solvent vehicl dilut prior treatment cell liquid test substanc ad directli test system dilut prior treatment fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl suspect chemic reaction test substanc compat surviv cell activ solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid test water unstabl substanc organ solvent free water water remov ad molecular siev exposur concentr criteria consid determin highest concentr cytotox solubl test system ph osmol cytotox determin metabol activ main experi indic cell integr growth degre confluenc viabl cell count mitot index determin cytotox solubl preliminari experi analys concentr cytotox occur concentr cover rang maximum toxic concentr separ factor radic time harvest highest concentr show signific reduct degre confluenc cell count mitot index greater mitot index indirect measur cytotox cytostat effect depend time treatment mitot index accept suspens cultur toxic measur cumbersom impract inform cell cycl kinet averag gener time agt supplementari inform agt averag reveal exist delai subpopul slight increas averag gener time substanti delai time optim yield aberr cytotox substanc maximum test concentr µl ml mg ml whichev lowest insolubl substanc toxic concentr lower insolubl concentr highest dose concentr limit solubl final cultur medium end treatment period case toxic occur higher lowest insolubl concentr advis test concentr visibl precipit assess solubl begin end treatment solubl chang exposur test system due presenc cell serum insolubl detect unaid ey precipit interfer score neg posit control concurr posit neg solvent vehicl control metabol activ includ experi metabol activ posit control chemic requir activ give mutagen respons posit control emploi elastogen exposur level expect give reproduc detect increas background demonstr sensit test system posit control concentr chosen effect clear immedi reveal ident code slide reader exampl posit control substanc includ gt tabl gt posit control substanc chemic class relat posit control chemic consid neg control consist solvent vehicl treatment medium treat treatment cultur includ harvest time addit untreat control histor control data demonstr deleteri mutagen effect induc chosen solvent procedur treatment test substanc prolifer cell treat test substanc presenc absenc metabol activ system treatment lymphocyt commenc hour mitogen stimul duplic cultur concentr strongli recommend neg solvent control cultur minim variat duplic cultur demonstr histor data accept singl cultur concentr gaseou volatil substanc test method seal cultur vessel cultur harvest time experi cell expos test substanc metabol activ hour sampl time equival normal cell cycl length begin treatment protocol neg result activ addit experi activ continu treatment sampl time equival normal cell cycl length chemic readili detect treatment sampl time longer cycl length neg result metabol activ conform case case basi case confirm neg result consid justif provid chromosom prepar cell cultur treat colcemid colchicin hour prior harvest cell cultur harvest process separ prepar chromosom chromosom prepar involv hypoton treatment cell fixat stain analysi slide includ posit neg control independ code microscop analysi fixat procedur result breakag proport metaphas cell loss chromosom cell score number centromer equal modal number ne cell type spread metaphas score concentr control equal divid duplic applic number reduc high number aberr observ purpos test detect structur chromosom aberr import record polyploidi endoredupl event data treatment result experiment unit cell percentag cell structur chromosom aberr evalu type structur chromosom aberr list number frequenc experiment control cultur gap record separ report gener includ total aberr frequenc concurr measur cytotox treat neg control cultur main aberr experi record individu cultur data provid addition data summaris tabular form requir verif clear posit respons equivoc result clarifi test prefer modif experiment condit confirm neg result discuss modif studi paramet extend rang condit assess consid follow experi studi paramet modifi includ concentr space metabol activ condit evalu interpret result criteria determin posit result concentr relat increas reproduc increas number cell chromosom aberr biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons increas number polyploid cell test substanc potenti inhibit mitot process induc numer chromosom aberr increas number cell endoredupl chromosom test substanc potenti inhibit cell cycl progress test substanc result meet criteria consid mutagen system experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result vitro chromosom aberr test test substanc induc structur chromosom aberr cultur mammalian somat cell neg result test condit test substanc induc chromosom aberr cultur mammalian somat cell report test report test report includ inform solvent vehicl justif choic vehicl solubl stabil test substanc solvent vehicl cell type sourc cell karyotyp featur suitabl cell type absenc mycoplasma applic inform cell cycl length sex blood donor blood separ lymphocyt mitogen number passag applic method mainten cell cultur applic modal number chromosom test condit ident metaphas arrest substanc concentr durat cell exposur rational select concentr number cultur includ cytotox data solubl limit composit media concentr applic concentr test substanc volum vehicl test substanc ad incub temperatur incub time durat treatment cell densiti seed type composit metabol activ system includ accept criteria posit neg control method slide prepar criteria score aberr number metaphas analys method measur toxic criteria studi posit neg equivoc result sign toxic degre confluenc cell cycl data cell count mitot index sign precipit data ph osmol treatment medium determin definit aberr includ gap number cell chromosom aberr type chromosom aberr separ treat control cultur ploidi dose respons relationship statist analys concurr neg solvent vehicl posit control data histor neg solvent vehicl posit control data rang mean standard deviat discuss result conclus refer evan cytolog method detect chemic mutagen chemic mutagen principl method detect vol hollaend ed plenum press york london pp ishid jr sofumi vitro chromosom aberr test chines hamster lung chl fibroblast cell cultur progress mutat research vol ashbi al ed elsevi scienc publish amsterdam york oxford pp gallowai armstrong reuben colman brown cannon bloom nakamura ahm duk rimpo margolin resnick anderson zeiger chromosom aberr sister chromat exchang chines hamster ovari cell evalu chemic environ molec mutagen suppl pp scott gallowai marshal ishid jr brusick ashbi myhr genotox extrem cultur condit report icpemc task group mutat re pp morita nagaki fukuda okumura clastogen low ph cultur mammalian cell mutat re pp am mccann yamasaki method detect carcinogen mutagen salmonella mammalian microsom mutagen test mutat re pp maron am revis method salmonella mutagen test mutat re pp natarajan tate van buul meijer de vogel cytogenet effect mutagen carcinogen activ mircrosom system vitro induct chromosom aberr sister chromatid exchang diethylnitrosamin den dimethylnitrosamin dmn cho cell presenc rat liver microsom mutat re pp matsuoka hayashi ishid jr chromosom aberr test chemic combin mix vitro mutat re pp elliot comb elcomb gatehous gibson mackai wolf report uk environment mutagen societi work parti altern aroclor induc vitro genotox assai mutagenesi pp matsushima sawamura hara sugimura safe substitut polychlorin biphenyl induc metabol activ system de serr fout bend philpot ed vitro metabol activ mutagenesi test elsevi north holland pp gallowai aardema ishid jr ivett kirkland morita mosesso sofumi report work group vitro test chromosom aberr mutat re pp richardson william allen amphlett chanter phillip analysi data vitro cytogenet assai statist evalu mutagen test data kirkland cd cambridg univers press cambridg pp soper gallowai replic flask vitro chromosom aberr assai cho cell mutat re pp krahn barski mccooei cho hgprt mutat assai evalu gase volatil liquid tice costa schaich ed genotox effect airborn agent york plenum pp zamora benson li brook evalu exposur system cell grown collagen gel detect highli volatil mutagen cho hgprt mutat assai environment mutagenesi pp lock huhl endoredupl chines hamster cell alpha radiat induc arrest mutat re pp huang chang trosko aphidicolin induc endoredupl chines hamster cell cancer re pp quot annex quot mutagen vivo mammalian bone marrow chromosom aberr test method method replic oecd tg mammalian bone marrow chromosom aberr test introduct mammalian vivo chromosom aberr test detect structur chromosom aberr induc test substanc bone marrow cell anim rodent structur chromosom aberr type chromosom chromatid increas polyploidi chemic potenti induc numer aberr major chemic mutagen induc aberr chromatid type chromosom type aberr occur chromosom mutat relat event human genet diseas substanti evid chromosom mutat relat event caus alter oncogen tumour suppress gene involv cancer human experiment system rodent arc routin test bone marrow target tissu test highli vascularis tissu popul rapidli cycl cell readili isol process speci target tissu subject method chromosom aberr test relev assess mutagen hazard consider factor vivo metabol pharmacokinet dna repair process vari speci tissu vivo test investig mutagen effect detect vitro test evid test substanc reactiv metabolit reach target tissu test gener introduct part definit chromatid type aberr structur chromosom damag express breakag singl chromatid breakag reunion chromatid chromosom type aberr structur chromosom damag express breakag breakag reunion chromatid ident site endoredupl process period dna replic nucleu mitosi start period result chromosom chromatid gap achromat lesion smaller width chromatid minimum misalign chromatid numer aberr chang number chromosom normal number characterist cell utilis polyploidi multipl haploid chromosom number diploid number structur aberr chang chromosom structur detect microscop examin metaphas stage cell divis observ delet fragment intrachang interchang principl test method anim expos test substanc rout exposur sacrif time treatment prior sacrific anim treat metaphas arrest agent colchicin colcemid chromosom prepar made bone marrow cell stain metaphas cell analys chromosom aberr descript test method prepar select anim speci rat mice chines hamster commonli mammalian speci commonli laboratori strain young healthi adult anim emploi commenc studi weight variat anim minim exce weight sex hous feed condit gener condit refer gener introduct part appli aim humid prepar anim healthi young adult anim randomli assign control treatment group cage arrang effect due cage placement minimis anim identifi uniqu anim acclim laboratori condit dai prepar dose solid test substanc dissolv suspend solvent vehicl dilut prior dose anim liquid test substanc dose directli dilut prior dose fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl produc toxic effect dose level suspect chemic reaction test substanc solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid control concurr posit neg solvent vehicl control includ sex test treatment test substanc anim control group handl ident manner anim treat group posit control produc structur aberr vivo exposur level expect give detect increas background posit control dose chosen effect clear immedi reveal ident code slide reader accept posit control administ rout test substanc sampl singl time chemic class relat posit control chemic consid exampl posit control substanc includ gt tabl gt neg control treat solvent vehicl treat treatment group includ sampl time accept inter anim variabl frequenc cell chromosom aberr histor control data singl sampl appli neg control time sampl time addit untreat control histor publish control data demonstr deleteri mutagen effect induc chosen solvent vehicl procedur number sex anim treat control group includ analys anim sex time studi data studi speci rout exposur demonstr substanti differ toxic sex test singl sex suffici human exposur chemic sex specif pharmaceut agent test perform anim sex treatment schedul test substanc prefer administ singl treatment test substanc administ split dose treatment dai separ hour facilit administ larg volum materi dose regimen scientif justifi sampl separ time treatment dai rodent sampl interv normal cell cycl length hour treatment time requir uptak metabol test substanc effect cell cycl kinet affect optimum time chromosom aberr detect sampl collect hour sampl time recommend dose regimen dai sampl time normal cell cycl length final treatment prior sacrific anim inject intraperiton dose metaphas arrest agent colcemid colchicin anim sampl interv mice interv approxim hour chines hamster interv approxim hour cell harvest bone marrow analys chromosom aberr dose level rang find studi perform suitabl data perform laboratori speci strain sex treatment regimen main studi toxic dose level sampl time dose level cover rang maximum toxic sampl time highest dose highest dose defin dose produc sign toxic higher dose level base dose regimen expect produc lethal substanc specif biolog activ low toxic dose hormon mitogen except dose set criteria evalu case case basi highest dose defin dose produc indic toxic bone marrow greater reduct mitot index limit test test dose level mg kg bodi weight singl treatment treatment dai produc observ toxic effect genotox expect base data structur relat substanc fulli studi dose level consid studi longer durat limit dose mg kg bodi weight dai treatment dai mg kig bodi weight dai treatment longer dai expect human exposur higher dose level limit test administr dose test substanc administ gavag stomach tube suitabl intub cannula intraperiton inject rout exposur accept justifi maximum volum liquid administ gavag inject time depend size test anim volum exce ml bodi weight volum higher justifi irrit corros substanc reveal exacerb effect higher concentr variabl test volum minimis adjust concentr ensur constant volum dose level chromosom prepar immedi sacrific bone marrow obtain expos hypoton solut fix cell spread slide stain analysi mitot index determin measur cytotox cell anim treat anim includ posit control untreat neg control anim cell analys anim number reduc high number aberr observ slide includ posit neg control independ code microscop analysi slide prepar procedur result breakag proport metaphas loss chromosom cell score number centromer equal number data treatment result individu anim data present tabular form experiment unit anim anim number cell score number aberr cell percentag cell structur chromosom aberr evalu type structur chromosom aberr list number frequenc treat control group gap record separ report gener includ total aberr frequenc evid differ respons sex data sex combin statist analysi evalu interpret result criteria determin posit result dose relat increas rel number cell chromosom aberr clear increas number cell aberr singl dose group singl sampl time biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons equivoc result clarifi test prefer modif experiment condit increas polyploidi test substanc potenti induc numer chromosom aberr increas endoredupl test substanc potenti inhibit cell cycl progress test substanc result meet criteria consid mutagen test experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number experi perform posit result vivo chromosom aberr test substanc induc chromosom aberr bone marrow speci test neg result test condit test substanc induc chromosom aberr bone marrow speci test likelihood test substanc metabolit reach gener circul specif target tissu system toxic discuss report test report test test report includ inform solvent vehicl justif choic vehicl solubl stabil test substanc solvent vehicl test anim speci strain number ag sex anim sourc hous condit diet individu weight anim start test includ bodi weight rang mean standard deviat group test condit posit neg vehicl solvent control data rang find studi conduct rational close level select detail test substanc prepar detail administr test substanc rational rout administr method verifi test substanc reach gener circul target tissu applic convers diet drink water test substanc concentr ppm actual dose mg kg bodi weight dai applic detail food water qualiti detail descript treatment sampl schedul method measur toxic ident metaphas arrest substanc concentr durat treatment method slide prepar criteria score aberr number cell analys anim criteria studi posit neg equivoc result sign toxic mitot index type number aberr separ anim total number aberr group mean standard deviat number cell aberr group mean standard deviat ploidi dose respons relationship statist analys concurr neg control data histor neg control data rang mean standard deviat concurr posit control data discuss result conclus refer adler cytogenet test mammal mutagen test practic approach venitt parri ed irl press oxford washington pp preston dean gallowai holden mcfee shelbi mammalian vivo cytogenet assai analysi chromosom aberr bone marrow cell mutat re pp richold chandlei ashbi gatehous bootman henderson vivo cytogenet assai kirkland ed basic mutagen test ukem recommend procedur ukem committe guidelin mutagen test report part revis cambridg univers press cambridg york port chester melbourn sydnei pp tice hayashi macgregaro anderson blakei holden kirsch volder oleson jr pacchierotti preston romagna shimada sutou vannier report work group vivo mammalian bone marrow chromosom aberr test mutat re pp fielder allen boobi botham doe esdail gatehous hodson walker morton kirkland richold report british toxicolog societi uk environment mutagen societi work group dose set vivo mutagen assai mutagenesi pp lovel anderson albanes amphlett clare ferguson richold papworth savag statist analysi vivo cytogenet assai ukem committe guidelin mutagen test report part iii statist evalu mutagen test data kirkland ed cambridg univers press cambridg pp lock huhl endoredupl chines hamster cell alpha radiat induc arrest mutat re pp huang chang trosko aphidicolin induc endoredupl chines hamster cell cancer re pp quot annex quot mutagen vivo mammalian erythrocyt micronucleu test method method replic oecd tg mammalian erythrocyt micronucleu test introduct mammalian vivo micronucleu test detect damag induc test substanc chromosom mitot apparatu erythroblast analysi erythrocyt sampl bone marrow peripher blood cell anim rodent purpos micronucleu test identifi substanc cytogenet damag result format micronuclei lag chromosom fragment chromosom bone marrow erythroblast develop polychromat erythrocyt main nucleu extrud micronucleu form remain anucl cytoplasm visualis micronuclei facilit cell lack main nucleu increas frequenc micronucl polychromat erythrocyt treat anim indic induc chromosom damag bone marrow rodent routin test polychromat erythrocyt produc tissu measur micronucl immatur polychromat erythrocyt peripher blood equal accept speci inabl spleen remov micronucl erythrocyt demonstr shown adequ sensit detect agent structur numer chromosom aberr micronuclei distinguish number criteria includ identif presenc absenc kinetochor centromer dna micronuclei frequenc micronucl immatur polychromat erythrocyt princip end point number matur normochromat erythrocyt peripher blood micronuclei number matur erythrocyt endpoint assai anim treat week mammalian vivo micronucleu test relev assess mutagen hazard consider factor vivo metabol pharmacokinet dna repair process vari speci tissu genet endpoint vivo assai investig mutagen effect detect vitro system evid test substanc reactiv metabolit reach target tissu test gener introduct part definit centromer kinetochor region chromosom spindl fiber cell divis allow orderli movement daughter chromosom pole daughter cell micronuclei small nuclei separ addit main nuclei cell produc telophas mitosi meiosi lag chromosom fragment chromosom normochromat erythrocyt matur erythrocyt lack ribosom distinguish immatur polychromat erythrocyt stain select ribosom polychromat erythrocyt immatur erythrocyt intermedi stage develop ribosom distinguish matur normochromat erythrocyt stain select ribosom principl test method anim expos test substanc rout bone marrow anim sacrif time treatment bone marrow extract prepar made stain peripher blood blood collect time treatment smear prepar made stain studi peripher blood time elaps exposur cell harvest prepar analys presenc micronuclei descript test method prepar select anim speci mice rat recommend bone marrow mammalian speci peripher blood mice recommend mammalian speci provid speci spleen remov micronucl erythrocyt speci shown adequ sensit detect agent structur numer chromosom aberr commonli laboratori strain young healthi anim emploi commenc studi weight variat anim minim exce weight sex hous feed condit gener condit refer gener introduct part appli aim humid prepar anim healthi young adult anim randomli assign control treatment group anim identifi uniqu anim acclim laboratori condit dai cage arrang effect due cage placement minimis prepar dose solid test substanc dissolv suspend solvent vehicl dilut prior dose anim liquid test substanc dose directli dilut prior dose fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl produc toxic effect dose level suspect chemic reaction test substanc solvent vehicl inclus support refer data indic compat recommend aqueou solvent vehicl consid control concurr posit neg solvent vehicl control includ sex test treatment test substanc anim control group handl ident manner anim treatment group posit control produc micronuclei vivo exposur level expect give detect increas background posit control dose chosen effect clear immedi reveal ident code slide reader accept posit control administ rout test substanc sampl singl time addit chemic class relat posit control chemic consid exampl posit control substanc includ gt tabl gt neg control treat solvent vehicl treat treatment group includ sampl time accept inter anim variabl frequenc cell micronuclei demonstr histor control data singl sampl appli neg control time sampl time addit untreat control histor publish control data demonstr deleteri mutagen effect induc chosen solvent vehicl peripher blood pre treatment sampl accept concurr neg control short peripher blood studi treatment result data expect rang histor control procedur number sex anim treat control group includ analys anim sex time studi data studi speci rout exposur demonstr substanti differ sex toxic test singl sex suffici human exposur chemic sex specif pharmaceut agent test perform anim sex treatment schedul standard treatment schedul treatment hour interv recommend sampl extend dose regimen accept long posit effect demonstr studi neg studi long toxic demonstr limit dose dose continu time sampl test substanc administ split dose treatment dai separ hour facilit administ larg volum materi test perform wai anim treat test substanc sampl bone marrow start earlier hour treatment extend hour treatment interv sampl sampl time earlier hour treatment justifi sampl peripher blood start earlier hour treatment interv sampl extend hour posit respons recognis sampl time addit sampl requir daili treatment treatment hour interv sampl collect hour final treatment bone marrow hour final treatment peripher blood sampl time addit relev dose level rang find studi perform suitabl data perform laboratori speci strain sex treatment regimen main studi toxic dose level sampl time dose level cover rang maximum toxic sampl time highest dose highest dose defin dose produc sign toxic higher dose level base dose regimen expect produc lethal substanc specif biolog activ low toxic dose hormon mitogen except dose set criteria evalu case case basi highest dose defin dose produc indic toxic bone marrow reduct proport immatur erythrocyt total erythrocyt bone marrow peripher blood limit test test dose level mg kg bodi weight singl treatment treatment dai produc observ toxic effect genotox expect base data structur relat substanc fulli studi dose level consid studi longer durat limit dose mg kg bodi weight dai treatment dai mg kg bodi weight dai treatment longer dai expect human exposur higher dose level limit test administr dose test substanc administ gavag stomach tube suitabl intub cannula intraperiton inject rout exposur accept justifi maximum volum liquid administ gavag inject time depend size test anim volum exce ml bodi weight volum higher justifi irrit corros substanc reveal exacerb effect higher concentr variabl test volum minimis adjust concentr ensur constant volum dose level bone marrow blood prepar bone marrow cell obtain femur tibia immedi sacrific commonli cell remov femur tibia prepar stain establish method peripher blood obtain tail vein blood vessel blood cell immedi stain supravit smear prepar made stain dna specif stain acridin orang hoechst pyronin elimin artifact dna specif stain advantag preclud convent stain giemsa addit system cellulos column remov nucleat cell provid system shown adequ work micronucleu prepar laboratori analysi proport immatur total immatur matur erythrocyt determin anim count total erythrocyt bone marrow erythrocyt peripher blood slide includ posit neg control independ code microscop analysi immatur erythrocyt anim score incid micronucl immatur erythrocyt addit inform obtain score matur erythrocyt micronuclei analys slide proport immatur erythrocyt total erythrocyt control anim treat continu week matur erythrocyt anim score incid micronuclei system autom analysi imag analysi flow cytometr analysi cell suspens accept altern manual evalu appropri justifi valid data treatment result individu anim data present tabular form experiment unit anim number immatur erythrocyt score number micronucl immatur erythrocyt number immatur total erythrocyt list separ anim analys anim treat continu week data matur erythrocyt collect proport immatur total erythrocyt consid applic percentag micronucl erythrocyt anim evid differ respons sex data sex combin statist analysi evalu interpret result criteria determin posit result dose relat increas number micronucl cell clear increas number micronucl cell singl dose group singl sampl time biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons equivoc result clarifi test prefer modif experiment condit test substanc result meet criteria consid mutagen test experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result micronucleu test substanc induc micronuclei result chromosom damag damag mitot apparatu erythroblast test speci neg result test condit test substanc produc micronuclei immatur erythrocyt test speci likelihood test substanc metabolit reach gener circul specif target tissu system toxic discuss report test report test report includ inform solvent vehicl justif choic vehicl solubl stabil test substanc solvent vehicl test anim speci strain number ag sex anim sourc hous condit diet individu weight anim start test includ bodi weight rang standard deviat group test condit posit neg vehicl solvent control data data rang find studi conduct rational dose level select detail test substanc prepar detail administr test substanc rational rout administr method verifi test substanc reach gener circul target tissu applic convers diet drink water test substanc concentr ppm actual dose mg kg bodi weight dai applic detail food water qualiti detail descript treatment sampl schedul method slide prepar method measur toxic criteria score micronucl immatur erythrocyt number cell analys anim criteria studi posit neg equivoc result sign toxic proport immatur erythrocyt total erythrocyt number micronucl immatur erythrocyt separ anim standard deviat micronucl immatur erythrocyt group dose respons relationship statist analys method appli concurr histor neg data concurr posit control data discuss result conclus refer heddl rapid vivo test chromosom damag mutat re pp schmid micronucleu test mutat re pp heddl salamon hite kirkhart mavournin macgregor newel induct micronuclei measur genotox mutat re pp mavournin blakei cimino salamon heddl vivo micronucleu assai mammalian bone marrow peripher blood report environment protect agenc gene tox program mutat re pp macgregor schlegel choi wehr micronuclei circul erythrocyt rapid screen chromosom damag routin toxic test mice develop scienc practic toxicolog ed hay schnell miya elsevi amsterdam pp macgregor heddl hite margolin ramel salamon tice wild guidelin conduct micronucleu assai mammalian bone marrow erythrocyt mutat re pp macgregor wehr henika shelbi vivo erythrocyt micronucleu test measur steadi state increas assai effici permit integr toxic studi fund appl toxicol pp hayashi morita kodama sofumi ishid jr micronucleu assai mous peripher blood reticulocyt acridin orang coat slide mutat re pp collabor studi group micronucleu test micronucleu test mous peripher blood erythrocyt acridim orang supravit stain summari report collabor studi csgmt jemm mm mutat re pp collabor studi group micronucleu test csgmt jemm mm mammalian mutagenesi studi group environment mutagen societi japan protocol recommend short term mous peripher blood micronucleu test mutagenesi pp hayashi tice macgregor anderson blackei kirsch volder oleson jr pacchicrotti romagna shimada sutou vannier vivo rodent erythrocyt micronucleu assai mutat re pp higashikuni sutou optim generalis sampl time doubl dose mous peripher micronucleu test mutagenesi pp fielder allen boobi botham doe esdail gatehous hodson walker morton kirkland rochold report british toxicolog societi uk environment mutagen societi work group dose set vivo mutagen assai mutagenesi pp hayashi sofumi ishid jr applic acridin orang fluoresc stain micronucleu test mutat re pp macgregor wehr langloi simpl fluoresc stain procedur micronuclei rna erythrocyt hoechst pyromin mutat re pp romagna staniforth autom bone marrow micronucleu test mutat re pp gollapudi mcfadden sampl size estim polychromat normochromat erythrocyt ratio bone marrow micronucleu test mutat re pp richold ashbi bootman chandlei gatehous henderson vivo cytogenet assai kirkland ed basic mutagen test ukem recommend procedur ukem committe guidelin mutagen test report part revis cambridg univers press cambridg york port chester melbourn sydnei pp lovel anderson albanes amphlett clare ferguson richold papworth savag static analysi vivo cytogenet assai kirkland ed statist evalu mutagen test data ukem committe guidelin mutagen test report part iii cambridg univers press cambridg york port chester melbourn sydnei pp quot annex quot mutagen revers mutat test bacteria method method replic oecd tg bacteri revers mutat test introduct bacteri revers mutat test amino acid requir strain salmonella typhimurium escherichia coli detect point mutat involv substitut addit delet dna base pair principl bacteri revers mutat test detect mutat revert mutat present test strain restor function capabl bacteria synthesis essenti amino acid revert bacteria detect abil grow absenc amino acid requir parent test strain point mutat human genet diseas substanti evid point mutat oncogen tumour suppressor gene somat cell involv tumour format human experiment anim bacteri revers mutat test rapid inexpens easi perform test strain featur make sensit detect mutat includ respons dna sequenc revers site increas cell permeabl larg molecul elimin dna repair system enhanc error prone dna repair process specif test strain provid inform type mutat induc genotox agent larg data base result wide varieti structur bacteri revers mutat test establish methodolog develop test chemic physico chemic properti includ volatil compound gener introduct part definit revers mutat test salmonella typhimurium escherichia coli detect mutat amino acid requir strain histidin tryptophan produc strain independ suppli amino acid base pair substitut mutagen agent base chang dna revers test chang occur site origin mutat site bacteri genom frameshift mutagen agent addit delet base pair dna chang read frame rna initi consider bacteri revers mutat test utilis prokaryot cell differ mammalian cell factor uptak metabol chromosom structur dna repair process test conduct vitro gener requir exogen sourc metabol activ vitro metabol activ system mimic mammalian vivo condit test provid direct inform mutagen carcinogen potenc substanc mammal bacteri revers mutat test commonli emploi initi screen genolox activ point mutat induc activ extens data base demonstr chemic posit test exhibit mutagen activ test exampl mutagen agent detect test reason shortcom ascrib specif natur endpoint detect differ metabol activ differ bioavail hand factor enhanc sensit bacteri revers mutat test lead overestim mutagen activ bacteri revers mutat test evalu class chemic highli bactericid compound antibiot thought interfer specif mammalian cell replic system topoisomeras inhibitor nucleosid analogu case mammalian mutat test compound posit test mammalian carcinogen correl absolut depend chemic class carcinogen detect test act genotox mechan mechan absent bacteri cell principl test method suspens bacteri cell expos test substanc presenc absenc exogen metabol activ system plate incorpor method suspens mix overlai agar plate immedi minim medium preincub method treatment mixtur incub mix overlai agar plate minim medium techniqu dai incub revert coloni count compar number spontan revert coloni solvent control plate procedur perform bacteri revers mutat test commonli plate incorpor method preincub method fluctuat method suspens method modif test gase vapour procedur method pertain primarili plate incorpor preincub method accept conduct experi metabol activ substanc detect effici preincub method substanc belong chemic class includ short chain aliphat nitrosamin dival metal aldehyd azo dye diazo compound pyrollizidin alkaloid allyl compound nitro compound recognis class mutagen detect standard procedur plate incorpor method preincub method regard special case strongli recommend altern procedur detect special case identifi exampl procedur detect azo dye diazo compound gase volatil chemic glycosid deviat standard procedur scientif justifi descript test method prepar bacteria fresh cultur bacteria grown late exponenti earli stationari phase growth approxim cell ml cultur late stationari phase essenti cultur experi high titr viabl bacteria titr demonstr histor control data growth curv assai determin viabl cell number plate experi recommend incub temperatur strain bacteria includ strain typhimurium ta ta ta ta ta ta shown reliabl reproduc respons laboratori typhimurium strain gc base pair primari revers site detect oxidis mutagen cross link agent hydrazin substanc detect coli wp strain typhimurium ta base pair primari revers site recommend combin strain typhimurium ta typhimurium ta ta ta typhimurium ta typhimurium ta coli wp uvra coli wp uvra pkm typhimurium ta order detect cross link mutagen prefer includ ta add dna repair profici strain coli coli wp coli wp pkm establish procedur stock cultur prepar marker verif storag amino acid requir growth demonstr frozen stock cultur prepar histidin typhimurium strain tryptophan coli strain phenotyp characterist similarli check presenc absenc factor plasmid ampicillin resist strain ta ta ta ta wp uvra wp uvra pkm ampicillin tetracyclin resist strain ta presenc characterist mutat rfa mutat typhimurium sensit crystal violet uvra mutat coli uvrb mutat typhimurium sensit ultra violet light strain yield spontan revert coloni plate count frequenc rang expect laboratori histor control data prefer rang report literatur medium minim agar vogel bonner minim medium glucos overlai agar histidin biotin tryptophan cell divis metabol activ bacteria expos test substanc presenc absenc metabol activ system commonli system cofactor supplement post mitochondri fraction prepar liver rodent treat enzym induc agent aroclor combin phenobarbiton naphthoflavon post mitochondri fraction concentr rang mix choic condit metabol activ system depend class chemic test case utilis concentr post mitochondri fraction azo dye diazo compound reduct metabol activ system test substanc prepar solid test substanc dissolv suspend solvent vehicl dilut prior treatment bacteria liquid test substanc ad directli test system dilut prior treatment fresh prepar emploi stabil data demonstr accept storag solvent vehicl suspect chemic reaction test substanc compat surviv bacteria activ solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid test water unstabl substanc organ solvent free water test condit test strain exposur concentr criteria consider determin highest amount test substanc cytotox solubl final treatment mixtur determin toxic insolubl preliminari experi cytotox detect reduct number revert coloni clear diminut background lawn degre surviv treat cultur cytotox substanc alter presenc metabol activ system insolubl assess precipit final mixtur actual test condit evid unaid ey recommend maximum test concentr solubl cytotox substanc mg plate µl plate cytotox substanc solubl mg plate µl plate concentr test insolubl final treatment mixtur test substanc cytotox mg plate µl plate test cytotox concentr precipit interfer score analys concentr test substanc approxim half log radic interv test point initi experi smaller interv concentr respons investig test concentr mg plate µl plate consid evalu substanc substanti amount potenti mutagen impur neg posit control concurr strain specif posit neg solvent vehicl control metabol activ includ assai posit control concentr demonstr effect perform assai select assai emploi metabol activ system posit control refer substanc select basi type bacteria strain substanc exampl suitabl posit control assai metabol activ gt tabl gt substanc suitabl posit control reduct metabol activ method gt tabl gt aminoanthracen sole indic efficaci mix aminoanthracen batch characteris mutagen requir metabol activ microsom enzym benzo pyren dimethylbenzanthracen substanc exampl strain specif posit control assai perform exogen metabol activ system gt tabl gt posit control refer substanc chemic class relat posit control chemic consid neg control consist solvent vehicl test substanc treat treatment group includ addit untreat control histor control data demonstr deleteri mutagen effect induc chosen solvent procedur plate incorpor method metabol activ ml ml test solut ml fresh bacteri cultur approxim viabl cell ml steril buffer mix ml overlai agar assai metabol activ ml metabol activ mixtur adequ amount post mitochondri fraction rang metabol activ mixtur mix overlai agar ml bacteria test substanc test solut content tube mix pour surfac minim agar plate overlai agar allow solidifi incub preincub method test substanc test solut preincub test strain approxim viabl cell steril buffer metabol activ system ml minut prior mix overlai agar pour surfac minim agar plate ml test substanc test solut ml bacteria ml mix steril buffer mix ml overlai agar tube aerat pre incub shaker adequ estim variat triplic plate dose level duplic plate accept scientif justifi occasion loss plate necessarili invalid assai gaseou volatil substanc test method seal vessel incub plate assai incub hour incub period number revert coloni plate count data treatment result data present number revert coloni plate number revert coloni neg solvent control untreat control posit control plate individu plate count number revert coloni plate standard deviat present test substanc posit neg untreat solvent control requir verif clear posit respons equivoc result clarifi test prefer modif experiment condit neg result confirm case case basi case confirm neg result consid justif provid modif studi paramet extend rang condit assess consid follow experi studi paramet modifi includ concentr space method treatment plate incorpor liquid pre incub metabol activ condit evalu interpret result criteria determin posit result concentr relat increas rang test reproduc increas concentr number revert coloni plate strain metabol activ system biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons test substanc result meet criteria consid mutagen test experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result bacteri revers mutat test substanc induc point mutat base substitut frameshift genom salmonella typhimurium escherichia coli neg result test condit test substanc mutagen test speci report test report test report includ inform solvent vehicl justif choic solvent vehicl solubl stabil test substanc solvent vehicl strain strain number cell cultur strain characterist test condit amount test substanc plate mg plate µl plate rational select dose number plate concentr media type composit metabol activ system includ accept criteria treatment procedur result sign toxic sign precipit individu plate count number revert coloni plate standard deviat dose respons relationship statist analys concurr neg solvent vehicl posit control data rang mean standard deviat histor neg solvent vehicl posit control data rang mean standard deviat discuss result conclus refer am mccann yamasaki method detect carcinogen mutagen salmonella mammalian microsom mutagen test mutat re pp maron am revis method salmonella mutagen test mutat re pp gatehous haworth cebula gock kier matsushima melcion nohmi venitt zeiger recommend perform bacteri mutat assai mutat re pp kier brusick auletta von hall brown simmon dunkel mccann mortelman prival rao rai salmonella typhimurium mammalian microsom assai report environment protect agenc gene tox program mutat re pp yahagi degawa seino matsushima nagao sugimura hashimoto mutagen carcinogen azo dye deriv cancer letter pp matsushima sugimura nagao yahagi shirai sawamura factor modul mutagen microbi test short term test system detect carcinogen ed norpoth garner springer berlin heidelberg york pp gatehous rowland wilcox callend foster bacteri mutat assai basic mutagen test ukem part revis ed kirkland cambridg univers press pp aeschach wolleb porchet liquid preincub mutagen test food food safeti pp green muriel bridg simplifi fluctuat test detect low level mutagen mutat re pp hubbard green gatehous bridg fluctuat test bacteria handbook mutagen test procedur edit ed kilbei legat nichol ramel elsevi amsterdam york oxford pp thompson melampi examin quantit suspens assai mutagenesi strain salmonella typhimurium environment mutagenesi pp araki noguchi kato matsushima improv method mutagen test gaseou compound ga sampl bag mutat re pp prival bell mitchel reipert vaughan mutagen benzidin benzidin congen dye select monoazo dye modifi salmonella assai mutat re pp zeiger anderson haworth lawlor mortelman salmonella mutagen test result test chemic environ mol mutagen pp simmon kauhanen tardiff mutagen activ chemic identifi drink water progress genet toxicolog scott bridg sobel ed elsevi amsterdam pp hugh simmon monteith claxton vaporis techniqu measur mutagen activ volatil organ chemic am salmonella assai environment mutagenesi pp matsushima matsumoto shirai sawamura sugimura mutagen natur occur carcinogen cycasin synthet methylazoxi methan conjug salmonella typhimurium cancer re pp tamura gold ferro luzzi am fecalas model activ dietari glycosid mutagen intestin flora proc natl acad sci usa pp wilcox naidoo wedd gatehous comparison salmonella typhimurium ta escherichia coli wp tester strain mutagenesi pp matsushima sawamura hara sugimura safe substitut polychlorin biphenyl induc metabol activ system vitro metabol activ mutagenesi test ed de serr al elsevi north holland pp elliot comb elcomb gatehous gibson mackai wolf altern aroclor induc vitro genotox assai mutagenesi pp maron katzenellenbogen am compat organ solvent salmonella microsom test mutat re pp claxton allen auletta mortelman nestmann zeiger guid salmonella typhimurium mammalian microsom test bacteri mutagen mutat re pp mahon green middleton mitchel robinson tweat analysi data microbi coloni assai ukem committe guidelin mutagen test part ii statist evalu mutagen test data ed kirkland cambridg univers press pp quot annex quot mutagen vitro mammalian cell gene mutat test method method replic oecd tg vitro mammalian cell gene mutat test introduct vitro mammalian cell gene mutat test detect gene mutat induc chemic substanc suitabl cell line includ mous lymphoma cell cho cho line chines hamster cell tk human lymphoblastoid cell cell line commonli genet endpoint measur mutat thymidin kinas tk hypoxanthin guanin phosphoribosyl transferas hprt transgen xanthin guanin phosphoribosyl transferas xprt tk hprt xprt mutat test detect spectra genet event autosom locat tk xprt detect genet event larg delet detect hprt locu chromosom vitro mammalian cell gene mutat test cultur establish cell line cell strain cell select basi growth abil cultur stabil spontan mutat frequenc test conduct vitro gener requir exogen sourc metabol activ metabol activ system mimic mammalian vivo condit care avoid condit lead result reflect intrins mutagen posit result reflect intrins mutagen aris ph osmol high level cytotox test screen mammalian mutagen carcinogen compound posit test mammalian carcinogen perfect correl test carcinogen correl depend chemic class increas evid carcinogen detect test act genotox mechan mechan absent bacteri cell gener introduct part definit forward mutat gene mutat parent type mutant form rise alter loss enzymat activ function encod protein base pair substitut mutagen substanc substitut base pair dna frameshift mutagen substanc addit delet singl multipl base pair dna molecul phenotyp express time period unalt gene product deplet newli mutat cell mutant frequenc number mutant cell observ divid number viabl cell rel total growth increas cell number time compar control popul cell calcul product suspens growth rel neg control time clone effici rel neg control rel suspens growth increas cell number express period rel neg control viabil clone effici treat cell time plate select condit express period surviv clone effici treat cell plate end treatment period surviv express relat surviv control cell popul prinicipl test method cell defici thymidin kinas tk due mutat tk tk resist cytotox effect pyrimidin analogu trifluorothymidin tft thymidin kinas profici cell sensit tft inhibit cellular metabol halt cell divis mutant cell prolifer presenc tft normal cell thymidin kinas similarli cell defici hprt xprt select resist thioguanin tg azaguanin ag properti test substanc consid carefulli base analogu compound relat select agent test mammalian cell gene mutat test suspect select toxic test substanc mutant mutant cell investig perform select system agent confirm test chemic structur relat select agent cell suspens monolay cultur expos test substanc metabol activ suitabl period time subcultur determin cytotox phenotyp express prior mutant select cytotox determin measur rel clone effici surviv rel total growth cultur treatment period treat cultur maintain growth medium suffici period time characterist select locu cell type optim phenotyp express induc mutat mutant frequenc determin seed number cell medium select agent detect mutant cell medium select agent determin clone effici viabil suitabl incub time coloni count mutant frequenc deriv number mutant coloni select medium number coloni select medium descript test method prepar cell varieti cell type test includ subclon cho cho tk cell cell type test demonstr sensit chemic mutagen high clone effici stabl spontan mutant frequenc cell check mycoplasma contamin contamin test design predetermin sensit power number cell cultur concentr test substanc reflect defin paramet minim number viabl cell surviv treatment stage test base spontan mutat frequenc gener guid cell number time invers spontan mutat frequenc recommend utilis cell adequ histor data cell system consist perform test media cultur condit cultur media incub condit cultur vessel temperatur concentr humid media chosen select system cell type test import cultur condit chosen ensur optim growth cell express period coloni form abil mutant mutant cell prepar cultur cell propag stock cultur seed cultur medium incub prior test cultur cleans pre exist mutant cell metabol activ cell expos test substanc presenc absenc metabol activ system commonli system cofactor supplement post mitochondri fraction prepar liver rodent treat enzym induc agent aroclor combin phenobarbiton naphthoflavon post mitochondri fraction concentr rang final test medium choic condit metabol activ system depend class chemic test case utilis concentr post mitochondri fraction number develop includ construct genet engin cell line express specif activ enzym provid potenti endogen activ choic cell line scientif justifi relev cytochrom isoenzym metabol test substanc test substanc prepar solid test substanc dissolv suspend solvent vehicl dilut prior treatment cell liquid test substanc ad directli test system dilut prior treatment fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl suspect chemic reaction test substanc compat surviv cell activ solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid test water unstabl substanc organ solvent free water water remov ad molecular siev exposur concentr criteria consid determin highest concentr cytotox solubl test system ph osmol cytotox determin metabol activ main experi indic cell integr growth rel clone effici surviv rel total growth determin cytotox solubl preliminari experi analys concentr cytotox concentr cover rang maximum toxic concentr level separ factor radic maximum concentr base cytotox result approxim rel surviv rel clone effici rel total growth cytotox substanc maximum test concentr mg ml µl ml whichev lowest insolubl substanc test limit solubl cultur condit evid insolubl determin final treatment medium cell expos assess solubl begin end treatment solubl chang exposur test system due presenc cell serum insolubl detect unaid ey precipit interfer score control concurr posit neg solvent vehicl control metabol activ includ experi metabol activ posit control chemic requir activ give mutagen respons exampl posit control substanc includ gt tabl gt posit control refer substanc laboratori histor data base bromo deoxyuridin ca einec refer substanc chemic class relat posit control chemic consid neg control consist solvent vehicl treatment medium treat treatment group includ addit untreat control histor control data demonstr deleteri mutagen effect induc chosen solvent procedur treatment test substanc prolifer cell expos test substanc metabol activ exposur suitabl period time hour effect exposur time extend cell cycl duplic singl treat cultur concentr test singl cultur number concentr increas ensur adequ number cultur analysi analys concentr duplic neg solvent control cultur gaseou volatil substanc test method seal cultur vessel measur surviv viabil mutant frequenc end exposur period cell wash cultur determin surviv express mutant phenotyp measur cytotox determin rel clone effici surviv rel total growth cultur initi treatment period locu defin minimum time requir optim phenotyp express newli induc mutant hprt xprt requir dai tk dai cell grown medium select agent determin number mutant clone effici measur viabil calcul mutant frequenc initi end express time plate select medium test substanc posit tk test coloni size perform test cultur highest posit concentr neg posit control test substanc neg tk test coloni size perform neg posit control studi tk tk coloni size perform data treatment result data includ cytotox viabil determin coloni count mutant frequenc treat control cultur case posit respons tk test coloni score criteria small larg coloni concentr test substanc highest posit concentr neg posit control molecular cytogenet natur larg small coloni mutant explor detail tk test coloni score criteria normal growth larg slow growth small coloni mutant cell suffer extens genet damag prolong doubl time form small coloni damag typic rang scale loss entir gene karyotyp visibl chromosom aberr induct small coloni mutant chemic induc gross chromosom aberr affect mutant cell grow rate similar parent cell form larg coloni surviv rel clone effici rel total growth mutant frequenc express number mutant cell number surviv cell individu cultur data provid addition data summaris tabular form requir verif clear posit respons equivoc result clarifi test prefer modif experiment condit neg result confirm case case basi case confirm neg result consid justif provid modif studi paramet extend rang condit assess consid follow experi equivoc neg result studi paramet modifi includ concentr space metabol activ condit evalu interpret result criteria determin posit result concentr relat increas reproduc increas mutant frequenc biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons test substanc result meet criteria consid mutagen system studi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result vitro mammalian cell gene mutat test test substanc induc gene mutat cultur mammalian cell posit concentr respons reproduc meaning neg result test condit test substanc induc gene mutat cultur mammalian cell report test report test report includ inform solvent vehicl justif choic vehicl solvent solubl stabil test substanc solvent vehicl cell type sourc cell number cell cultur number cell passag applic method mainten cell cultur applic absenc mycoplasma test condit rational select concentr number cultur includ cytotox data solubl limit composit media concentr concentr test substanc volum vehicl test substanc ad incub temperatur incub time durat treatment cell densiti treatment type composit metabol activ system includ accept criteria posit neg control length express period includ number cell seed subcultur feed schedul select agent criteria test posit neg equivoc method enumer number viabl mutant cell definit coloni size type consid includ criteria small larg coloni result sign toxic sign precipit data ph osmol exposur test substanc determin coloni size score neg posit control laboratori adequaci detect small coloni mutant tk system dose respons relationship statist analys concurr neg solvent vehicl posit control data histor neg solvent vehicl posit control data rang mean standard deviat mutant frequenc discuss result conclus refer moor demarini deserr tindal ed banburi report mammalian cell mutagenesi cold spring harbor laboratori york chu mall mammalian cell genet ii chemic induct specif locu mutat chines hamster cell vitro proc natl acad sci usa pp liber thilli mutat assai thymidin kinas locu diploid human lymphoblast mutat re pp moor harington brock doerr dearfield differenti mutant quantit mous lymphoma tk cho hgprt loci mutagenesi pp aaron stankowski jr comparison xprt cho hprt assai evalu drug candid mutat re pp aaron bolcsfoldi glatt moor nishi stankowski jr theiss thompson mammalian cell gene mutat assai work group report report intern workshop standardis genotox test procedur mutat re pp scott gallowai marshal ishid brusick ashbi myhr genotox extrem cultur condit report icpemc task group mutat re pp clive mccuen spector piper mavournin specif gene mutat cell cultur report environment protect agenc gene tox program mutat re pp li gupta heflich wasson review analysi chines hamster ovari hypoxanthin guanin phosphoribosyl transferas system determin mutagen chemic agent report phase iii environment protect agenc gene tox program mutat re pp li carver choi hsie gupta lovedai óneill riddl stankowski jr yang guid perform chines hamster ovari cell hypoxanthin guanin phosphoribosyl transferas gene mutat assai mutat re pp liber yandel comparison mutat induct tk hprt loci human lymphoblastoid cell quantit differ due addit class mutat autosom tk locu mutat re pp stankowski jr tindal hsie quantit molecular analys ethyl methanosulphon icr induc molecular analys ethyl methanosulphon icr induc mutat cell mutat re pp turner batson clive procedur tk tk mous lymphoma cell mutagen assai kilbei al ed handbook mutagen test procedur elsevi scienc publish york pp arlett smith clark green cole mcgregor asquith mammalian cell gene mutat assai base coloni format statist evalu mutagen test data kirkland ed cambirdg univers press pp abbondandolo bonatti corti fiorio loprieno mazzaccoro induct thioguanin resist mutant chines hamster cell mous liver microsom activ dimethylnitrosamin mutat re pp am mccann yamasaki method detect carcinogen mutagen salmonella mammalian microsom mutagen test mutat re pp clive johnson spector batson brown valid characteris tk mous lymphoma mutagen assai system mutat re pp maron am revis method salmonella mutagen test mutat re pp elliott comb elcomb gatehous gibson mackai wolf altern aroclor induc vitro genotox assai mutagenesi pp matsushima sawamura hara sugimura safe substitut polycholrin biphenyl induc metabol activ system vitro metabol activ mutagenesi test de serr fout bend philpot ed elsevi north holland pp krahn barski mccooei cho hgprt mutat assai evalu gase volatil liquid ticc costa schaich ed genotox effect airborn agent york plenum pp zamora benson li brook evalu exposur system cell grown collagen gel detect highli volatil mutagen cho hgprt mutat assai environment mutagenesi pp appleg moor broder burrel hozier molecular dissect mutat heterozyg thymidin kinas locu mous lymphoma cell proc natl acad sci usa pp moor clive hozier howard batson turner sawyer analysi trifluoronthymidin resist tft mutant tk mous lymphoma cell mutat re pp yandel dryja molecular genet analysi recess mutat heterozyg autosom locu human cell mutat re pp moor doerr comparison chromosom aberr frequenc small coloni tk defici mutant frequenc tk mous lymphoma cell mutagenesi pp quot annex quot mammalian spermatogoni chromosom aberr test method method replic oecd tg mammalian spermatogoni chromosom aberr test introduct purpos vivo mammalian spermatogoni chromosom aberr test identifi substanc structur chromosom aberr mammalian spermatogoni cell structur aberr type chromosom chromatid major chemic mutagen induc aberr chromatid type chromosom type aberr occur method design measur numer aberr routin purpos chromosom mutat relat event human genet diseas test measur chromosom event spermatogoni germ cell expect predict induct inherit mutat germ cell rodent routin test vivo cytogenet test detect chromosom aberr spermatogoni mitos target cell subject method detect chromatid type aberr spermatogoni cell mitot cell divis treatment examin lesion lost subsequ cell divis addit inform treat spermatogoni stem cell obtain meiotic chromosom anaylsi chromosom type aberr diakinesi metaphas treat cell spermatocyt vivo test design investig somat cell mutagen activ germ cell addit spermatogoni test relev assess mutagen hazard consider factor vivo metabol pharmacokinet dna repair process number gener spermatogonia present testi spectrum sensit chemic treatment aberr detect repres aggreg respons treat spermatogoni cell popul numer differenti spermatogoni cell predomin depend posit testi gener spermatogonia expos gener circul physic physiolog sertoli cell barrier blood testi barrier evid test substanc reactiv metabolit reach target tissu test gener introduct part definit chromatid type aberr structur chromosom damag express breakag singl chromatid breakag reunion chromatid chromosom type aberr structur chromosom damag express breakag breakag reunion chromatid ident site gap achromat lesion smaller width chromatid minimum misalign chromatid numer aberr chang number chromosom normal number characterist anim utilis polyploidi multipl haploid chromosom number diploid number structur aberr chang chromosom structur detect microscop examin metaphas stage cell divis observ delet intrachang interchang principl test method anim expos test substanc rout exposur sacrif time treatment prior sacrific anim treat metaphas arrest substanc colchicin colcemid chromosom prepar made germ cell stain metaphas cell analys chromosom aberr descript test method prepar select anim speci male chines hamster mice commonli male mammalian speci commonli laboratori strain young healthi adult anim emploi commenc studi weight variat anim minim exce weight hous feed condit gener condit refer gener introduct part appli aim humid prepar anim healthi young adult male randomli assign control treatment group cage arrang effect due cage placement minimis anim identifi uniqu anim acclim laboratori condit dai prior start studi prepar dose solid test substanc dissolv suspend solvent vehicl dilut prior dose anim liquid test substanc dose directli dilut prior dose fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl produc toxic effect dose level suspect chemic reaction test substanc solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid control concurr posit neg solvent vehicl control includ test treatment test substanc anim control group handl ident manner anim treat group posit control produc structur chromosom aberr vivo spermatogoni cell administ exposur level expect give detect increas background posit control dose chosen effect clear immedi reveal ident code slide reader accept posit control administ rout test substanc sampl singl time addit chemic class relat posit control chemic consid exampl posit control substanc includ gt tabl gt neg control treat solvent vehicl treat treatment group includ sampl time accept inter anim variabl frequenc cell chromosom aberr demonstr histor control data addit untreat control histor publish control data demonstr deleteri mutagen effect induc chosen solvent vehicl procedur number anim treat control group includ analys male treatment schedul test substanc prefer administ singl treatment treatment test substanc administ split dose treatment dai separ hour facilit administ larg volum materi dose regimen scientif justifi highest dose group sampl time treatment cell cycl kinet influenc test substanc earli late sampl time hour treatment dose highest dose sampl time hour cell cycl length treatment sampl time detect effect addit sampl time case chemic induc chromosom lag exert independ effect earlier sampl time appropri repeat treatment schedul identifi case case basi repeat treatment schedul anim sacrif hour cell cycl length treatment addit sampl time prior sacrific anim inject intraperiton dose metaphas arrest substanc colcemid colchocin anim sampl interv mice interv approxim hour chines hamster interv approxim hour dose level rang find studi perform suitabl data perform laboratori speci strain treatment regimen main studi toxic dose level sampl time dose level cover rang maximum toxic sampl time highest dose highest dose defin dose produc sign toxic higher dose level base dose regimen expect produc lethal substanc specif biolog activ low toxic dose hormon mitogen except dose set criteria evalu case case basi highest dose defin dose produc indic toxic spermatogoni cell reduct ratio spermatogoni mitos meiotic metaphas reduct exce limit test test dose level mg kg bodi weight dai singl treatment treatment dai produc observ toxic effect genotox expect base data structur relat substanc full studi dose level consid expect human exposur higher dose level limit test administr dose test substanc administ gavag stomach tube suitabl intub cannula intraperiton inject rout exposur accept justifi maximum volum liquid administ gavag inject time depend size test anim volum exce ml bodi weight volum higher justifi irrit corros substanc reveal exacerb effect higher concentr variabl test volum minimis adjust concentr ensur constant volum dose level chromosom prepar immedi sacrific cell suspens obtain test expos hypoton solut fix cell spread slide stain analysi anim spread metaphas analys minimum metaphas group number reduc high number aberr observ slide includ posit neg control independ code microscop analysi fixat procedur result breakag proport metaphas loss chromosom cell score number centromer equal number data treatment result individu anim data present tabular form experiment unit anim individu anim number cell structur chromosom aberr number chromosom aberr cell evalu type structur chromosom aberr list number frequenc treat control group gap record separ report gener includ total aberr frequenc mitosi meiosi observ ratio spermatogoni mitos meiotic metaphas determin measur cytotox treat neg control anim total sampl divid cell anim establish cytotox effect mitosi observ mitosi index determin cell anim evalu interpret result criteria determin posit result dose relat increas rel number cell chromosom aberr clear increas number cell aberr singl dose singl sampl time biolog relev result consid statist method aid evalu test result statist signific determin factor posit respons equivoc result clarifi test prefer modif experiment condit test substanc result meet criteria consid mutagen test experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result vivo spermatogoni chromosom aberr test test substanc induc structur chromosom aberr germ cell speci test neg result test condit test substanc induc chromosom aberr germ cell speci test likelihood test substanc metabolit reach target tissu discuss report test report test report includ inform solvent vehicl justif choic vehicl solubl stabil test substanc solvent vehicl test anim speci strain number ag anim sourc hous condit diet individu weight anim start test includ bodi weight rang standard deviat group test condit data rang find studi conclud rational dose level select rational rout administr detail test substanc prepar detail administr test substanc rational sacrific time convers diet drink water test substanc concentr ppm actual dose mg kg bodi weight dai applic detail food water qualiti detail descript treatment sampl schedul method measur toxic ident metaphas arrest substanc concentr durat treatment method slide prepar criteria score aberr number cell analys anim criteria studi posit neg equivoc result sign toxic mitot index ratio spermatogoni mitos cell meiotic metaphas type number aberr separ anim total number aberr group number cell aberr group dose respons relationship statist analys concurr neg control data histor neg control data rang mean standard deviat concurr posit control data ploidi discuss result conclus refer adler clastogen potenti mous spermatogonia chemic mutagen relat cell cycl specif genet toxicolog environment chemic part genet effect appli mutagenesi ramel lambert magnusson ed liss york pp adler cytogenet test mammal mutagen test practic approach ed venitt parri irl press oxford washington dc pp evan breckon ford air dry method meiotic prepar mammalian test cytogenet cell genet pp richold ashbi chandlei gatehous henderson vivo cytogenet assai kirkland ed basic mutagen test ukem recommend procedur ukem subcommitte guidelin mutagen test report part revis cambridg univers press cambridg york port chester melbourn sydnei pp yamamoto kikuchi method prepar mammalian spermatogoni chromosom mutat re pp adler shelbi bootman favor generoso pacchierotti shibuya tanaka intern workshop standardis genotox test procedur summari report work group mammalian germ cell test mutat re pp fielder allen boobi botham doe esdail gatehous hodson walker morton kirkland richold report british toxicolog societi uk environment mutagen societi work group dose set vivo mutagen assai mutagenesi pp lovel anderson albanes amphlett clarc ferguson richold papworth savag statist analysi vivo cytogenet assai kirkland ed statist evalu mutagen test data ukem subcommitte guidelin mutagen test report part iii cambridg univers press cambridg york port chester melbourn sydnei pp quot annex quot unschedul dna synthesi ud test mammalian liver cell vivo method method replic oecd tg unschedul dna synthesi ud test mammalian liver cell vivo introduct purpos unschedul dna synthesi ud test mammalian liver cell vivo identifi test substanc induc dna repair liver cell treat anim vivo test method investig genotox effect chemic liver end point measur indic dna damag subsequ repair liver cell liver major site metabol absorb compound site measur dna damag vivo evid test substanc reach target tissu test end point unschedul dna synthesi ud measur determin uptak label nucleosid cell undergo schedul phase dna synthesi wide techniqu determin uptak tritium label thymidin tdr autoradiographi rat liver prefer vivo ud test tissu liver subject method detect ud respons depend number dna base excis replac site damag ud test valuabl detect substanc induc longpatch repair base contrast shortpatch repair base detect lower sensit mutagen event result repair misrepair misrepl dna lesion extent ud respons indic fidel repair process addit mutagen react dna dna damag repair excis repair process lack specif inform mutagen activ provid ud test compens potenti sensit end point measur genom gener introduct part definit cell repair net nuclear grain nng higher preset justifi laboratori conduct test net nuclear grain nng quantit measur ud activ cell autoradiograph ud test calcul subtract averag number cytoplasm grain nucleu equival cytoplasm area cg number nuclear grain ng nng ng cg nng count calcul individu cell pool cell cultur parallel cultur unschedul dna synthesi ud dna repair synthesi excis remov stretch dna region damag induc chemic substanc physic agent principl test method ud test mammalian liver cell vivo dna repair synthesi excis remov stretch dna region damag induc chemic substanc physic agent test base incorpor tdr dna liver cell low frequenc cell phase cell cycl uptak tdr determin autoradiographi techniqu suscept interfer phase cell liquid scintil count descript method prepar select anim speci rat commonli mammalian speci commonli laboratori strain young healthi adult anim emploi commenc studi weight variat anim minim exce weight sex hous feed condit gener condit refer gener introduct part appli aim humid prepar anim healthi young adult anim randomli assign control treatment group cage arrang effect due cage placement minimis anim identifi uniqu cage dai prior start studi acclimatis laboratori condit test substanc prepar solid test substanc dissolv suspend solvent vehicl dilut prior dose anim liquid test substanc dose directli dilut prior dose fresh prepar test substanc emploi stabil data demonstr accept storag test condit solvent vehicl solvent vehicl produc toxic effect dose level suspect chemic reaction test substanc solvent vehicl inclus support data indic compat recommend aqueou solvent vehicl consid control concurr posit neg control solvent vehicl includ independ perform part experi treatment test substanc anim control group handl ident manner anim treat group posit control substanc produc ud administ exposur level expect give detect increas background posit control need metabol activ dose elicit moder respons dose chosen effect clear immedi reveal ident code slide reader exampl posit control substanc includ gt tabl gt posit control substanc accept posit control administ rout test substanc procedur number sex anim adequ number anim account natur biolog variat test respons number anim analys anim group signific histor databas accumul anim requir concurr neg posit control group time studi data studi speci rout exposur demonstr substanti differ toxic sex test singl sex prefer male suffici human exposur chemic sex specif pharmaceut agent test perform anim sex treatment schedul test substanc gener administ singl treatment dose level dose level highest dose defin dose produc sign toxic higher dose level base dose regimen expect produc lethal gener lower dose high dose substanc specif biolog activ low toxic dose hormon mitogen except dose set criteria evalu case case basi rang find studi perform suitabl data perform laboratori speci strain sex treatment regimen main studi highest dose defin dose produc indic toxic liver pyknot nuclei limit test test dose level mg kg bodi weight appli singl treatment treatment dai produc observ toxic effect genotox expect base data structur relat substanc full studi expect human exposur higher dose level limit test administr dose test substanc administ gavag stomach tube suitabl intub cannula rout exposur accept justifi intraperiton rout recommend expos liver directli test substanc circulatori system maximum volum liquid administ gavag inject time depend size test anim volum exce ml bodi weight volum higher justifi irrit corros substanc reveal exacerb effect higher concentr variabl test volum minimis adjust concentr ensur constant volum dose level prepar liver cell liver cell prepar treat anim hour dose addit earlier sampl time hour post treatment gener clear posit respons hour altern sampl time justifi basi toxicokinet data short term cultur mammalian liver cell establish perfus liver situ collagenas allow freshli dissoci liver cell attach suitabl surfac liver cell neg control anim viabil determin ud freshli isol mammalian liver cell incub medium tdr length time hour end incub period medium remov cell incub medium excess unlabel thymidin diminish unincorpor radioact cold chase cell rins fix dri prolong incub time cold chase slide dip autoradiograph emuls expos dark refriger dai develop stain expos silver grain count slide prepar anim analysi slide prepar suffici cell normal morpholog permit meaning assess ud prepar examin microscop sign overt cytox pyknosi reduc level radiolabel slide code grain count cell score anim slide score cell anim justifi grain count score phase nuclei proport phase cell record amount tdr incorpor nuclei cytoplasm morpholog normal cell evidenc deposit silver grain determin suitabl method data treatment result individu slide anim data provid addition data summaris tabular form net nuclear grain nng count calcul cell anim dose time subtract cg count ng count cell repair count criteria defin cell repair justifi base histor concurr neg control data numer result evalu statist method statist test select justifi prior conduct studi evalu interpret result gt tabl gt biolog relev data consid paramet inter anim variat dose respons relationship cytotox account statist method aid evalu test result statist signific determin factor posit respons experi give posit neg result rare case data set preclud make definit judgement activ test substanc result remain equivoc question number time experi repeat posit result ud test mammalian liver cell vivo test substanc induc dna damag mammalian liver cell vivo repair unschedul dna synthesi vitro neg result test condit test substanc induc dna damag detect test likelihood test substanc reach gener circul specif target tissu system toxic discuss report test report test report includ inform solvent vehicl justif choic vehicl solubl stabil test substanc solvent vehicl test anim speci strain number ag sex anim sourc hous condit diet individu weight anim start test includ bodi weight rang standard deviat group test condit posit neg vehicl solvent control data rang find studi conduct rational dose level select detail test substanc prepar detail administr test substanc rational rout administr method verifi test agent reach gener circul target tissu applic convers diet drink water test substanc concentr ppm actual dose mg kg bodi weight dai applic detail food water qualiti detail descript treatment sampl schedul method measur toxic method liver cell prepar cultur autoradiograph techniqu number slide prepar number cell score evalu criteria criteria studi posit neg equivoc result individu slide anim group valu nuclear grain cytoplasm grain net nuclear grain dose respons relationship statist evalu sign toxic concurr neg solvent vehicl posit control data histor neg solvent vehicl posit control data rang mean standard deviat number cell repair determin number phase cell determin viabil cell discuss result conclus refer ashbi lefevr burlinson penman assess vivo rat hepatocyt dna repair assai mutat re pp butterworth ashbi bermudez casciano mirsali probst william protocol guid vivo rat hepatocyt dna repair assai mutat re pp kennelli water ashbi lefevr burlinson benford dean mitchel de vivo rat liver ud assai kirkland fox ed supplementari mutagen test ukem recommend procedur ukem subcommitte guidelin mutagen test report part ii revis cambridg univers press cambridg york port chester melbourn sydnei pp madl dean andrac brambilla burlinson doolittl furihata hertner mcqueen mori recommend perform ud test vitro vivo mutat re pp fautz hussain efstathi hechenberg freudl assess relat initi viabil attach freshli isol rat hepatocyt vivo vitro dna repair assai ud mutat re pp mirsali tyson butterworth detect genotox carcinogen vivo vitro hepalocyt dna repair assai environ mutagen pp quot annex sv farligt farligt kan ge lungskador vid förtäring flytand ämnen och beredningar som på grund av sin låga viskositet utgör en fara för människa vid aspir för ämnen och beredningar som innehål alifatiska alicykliska och aromatiska kolväten en total koncentr av eller mer och har en flödestid mindr än sekund uppmätt med en mm utloppsbägar enligt iso eller har en kinematisk viskositet lägre än vid uppmätt med en kalibrerad kapillärviskosimet av gla enligt iso och iso eller har en kinematisk viskositet lägre än vid bestämd från rotationsviskosimetri enligt iso ämnen och beredningar som uppfyl dessa kriteri behöver dock int klassificera om de har en genomsnitlig ytspänning högre än mn vid uppmätt med du noüytensiomet eller enligt de testmetod som finn beskrivna bilaga del för ämnen och beredningar baserat på praktiska erfarenhet från människa concern es version concern da version concern de version concern el version concern en version concern fr version concern version concern nl version concern pt version concern fi version fi ihon tulehtuminen seuraava vaaraa osoittava lauseka määräytyi alla esiteltävien perusteitten mukaan ärsyttää ihoa aineet ja valmisteet aiheuttavat ihon merkittävän tulehtumisen enintään neljän tunnin altistuksessa määritettynä kanilla liitteessä mainitulla ihoärsytstestillä tulehdu kestää vähintään tuntia ihon tulehdu merkittävää jo punoituksen ja ruvenmuodostuksen tai turvotuksen voimakkuutta kuvaavien lukuarvojen keskiarvo laskettuna kaikista koe eläimistä vähintään tai kun liitteessä tarkoitettua testiä täydennetti käyttämällä kolmea koe eläintä vähintään kahden koe eläimen ihon punoituksen ja ruvenmuodostuksen tai turvotuksen voimakkuutta kuvaavien lukuarvojen keskiarvo jokaisel koe eläimel laskettuna erikseen vähintään kummassakin tapauksessa keskiarvojen lasekmiseen käytettävä kaikkia niitä lukuarvoja jotka saadaan arvioitaessa vaikutusta tunnin tunnin ja tunnin välein tulehdusta pidetään myö merkittävänä jo ihon tulehtuminen jatkuu ainakin kahdella koe eläimellä havainnointiajan päättymiseen asti erityiset vaikutukset kuten esimerkiksi hyperplasia hilseileminen värin muutokset halkeamat ruvet ja karvojenlähtö otettava huomioon tähän liittyviä tietoja voidaan saada myö eläimillä tehtävistä ei akuuttisista altistuskokeista katso lauseketta koskevat huomautukset jaksossa vaikutuksia pidetään merkittävinä jo ne vastaavat edellä kuvattuja vaikutuksia aineet ja valmisteet jotka aiheuttavat ihmisillä merkittävää ihotulehdusta kun kosketu ollut välitön jatkuva tai toistuva orgaaniset peroksidit paitsi jo olemassa näyttöä siitä että tällaista vaikutusta ei ol tuntoharha quot paresthesia quot pyretroiditorjunta aineen ihokosketuksen aiheuttamaa tuntoharhaa ihmisessä ei pidetä ärsytysvaikutuksena joka oikeuttaisi luokituksen xi lauseketta kuitenkin sovellettava aineisiin joilla tällainen vaikutu concern es version concern da version concern de version concern el version concern en version concern fr version concern version concern nl version concern pt version concern sv version point safeti phrase substanc prepar de bei berührung mit der haut sofort mit viel abwaschen vom herstel anzugeben anwendungsbereich sehr giftig giftig oder ätzend stoff und zubereitungen verwendung obligatorisch für sehr giftig stoff und zubereitungen empfohlen für sonstig obengenannt stoff und zubereitungen inbesonder wenn wasser nicht die geeignet spülflüssigkeit ist empfohlen für ätzend stoff und zubereitungen die die allgemein öffentlichkeit abgegeben werden concern es version concern da version concern el version concern en version concern fr version concern version concern nl version concern pt version concern fi version concern sv version fi ei saa tyhjentää viemäriin soveltamisala erittäin helposti syttyvät tai helposti syttyvät veteen sekoittumattomat nesteet erittäin myrkylliset tai myrkylliset aineet ja valmisteet ympäristöll vaaralliset aineet käytön perusteet pakollinen yleisessä kulutuksessa todennäkoisesti käytettävil ympäristöll vaarallisil ja tunnuksella luokitelluil aineil jollei kyseessä ol aineen tarkoitettu käyttö suositeltava yleisessä kulutuksessa todennäkoisesti käytettävil muill edellä mainituil aineil tai valmisteil jollei kyseessä ol kemikaalin tarkoitettu käyttö concern es version concern da version concern de version concern el version concern en version concern fr version concern version concern nl version concern pt version concern sv version annex quot annex ix part provis relat child proof fasten addit provis articl direct contain capac substanc present aspir hazard xn classifi label paragraph annex vi direct except substanc market form aerosol contain fit seal sprai attach fit child proof fasten reclos packag child proof fasten reclos packag compli iso standard juli edit relat quot child resist packag requir method test reclos packag quot adopt intern standard organis iso reclos packag child proof fasten reclos packag compli cen standard en march edit relat quot packag child resist packag requir test procedur reclos packag pharmaceut product quot adopt european committe standardis cen note evid conform standard certifi laboratori conform european standard seri en specif case obviou packag suffici safe children access content tool test perform case suffici ground doubt secur closur child nation author person respons put product market give certif laboratori state type closur test iso cen standard refer closur test found conform standard refer part provis relat tactil warn devic technic specif tactil warn devic conform en iso standard edit relat quot packag tactil warn danger requir quot quot