commiss direct ec septemb 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 commiss direct ec articl thereof annex direct eec list danger substanc particular classif label procedur respect substanc present scientif technic knowledg shown list danger substanc annex adapt supplement annex direct eec lai method determin physico chemic properti toxic ecotox substanc prepar adapt annex technic progress 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 entri annex direct replac entri annex direct eec entri annex ii direct insert annex direct eec annex amend text annex iii iii iii direct ad part annex direct eec text annex iii direct ad part annex direct eec articl member state bring forc law regul administr provis compli direct octob latest forthwith inform commiss thereof member state adopt provis refer direct accompani refer occas offici public member state determin refer made articl direct enter forc dai public offici journal european commun articl direct address member state brussel septemb commiss ritt bjerregaard member commiss oj oj anexo bilag anhang annex annex allegato bijlag anexo liit bilaga gt refer graphic gt anexo ii bilag ii anhang ii ii annex ii annex ii allegato ii bijlag ii anexo ii liit ii bilaga ii gt refer graphic gt annex iii number averag molecular weight molecular weight distribut polym method gel permeat chromatograph method replic oecd tg fundament principl technic inform refer introduct properti polym vari imposs describ singl method set precis condit separ evalu cover eventu specif occur separ polym complex polym system amen gel permeat chromatographi gpc gpc practic molecular weight determin mean method annex case full detail justif method method base din standard detail inform carri experi evalu data found din standard case modif experiment condit justifi standard fulli referenc method polystyren sampl polydispers calibr modifi suitabl polym water solubl long chain branch polym definit unit number averag molecular weight mn weight averag molecular weight mw determin equat mn gt num gt ói lnhi gt den gt ói ln gt num gt gt den gt mi mw gt num gt ói lnhixmi gt den gt ói lnhi level detector signal baselin retent volum vi mi molecular weight polym fraction retent volum vi number data point breadth molecular weight distribut measur dispers system ratio mw mn refer substanc gpc rel method calibr undertaken narrowli distribut linearli construct polystyren standard averag molecular weight mn mw molecular weight distribut calibr curv determin molecular weight unknown sampl condit separ sampl standard select ident manner determin relationship molecular weight elut volum valid specif condit experi condit includ temperatur solvent solvent mixtur chromatographi condit separ column system column molecular weight sampl determin rel valu polystyren equival molecular weight mean depend structur chemic differ sampl standard molecular weight deviat absolut valu greater lesser degre standard polyethylen glycol polyethylen oxid polymethyl methacryl polyacryl acid reason state principl test method molecular weight distribut sampl averag molecular weight mn mw determin gpc gpc special type liquid chromatographi sampl separ hydrodynam volum individu constitu separ effect sampl pass column fill porou materi typic organ gel small molecul penetr pore larg molecul exclud path larg molecul shorter elut medium size molecul penetr pore elut smallest molecul hydrodynam radiu smaller pore gel penetr pore elut ideal situat separ govern size molecular speci practic difficult avoid absorpt effect interf uneven column pack dead volum worsen situat detect effect refract index uv absorpt yield simpl distribut curv attribut actual molecular weight valu curv calibr column pass polym molecular weight ideal broadli similar structur polystyren standard typic gaussian curv result distort small tail low molecular weight side vertic axi indic quantiti weight molecular weight speci elut horizont axi log molecular weight qualiti criteria repeat rel standard deviat rsd elut volum requir repeat analysi ensur correct intern standard chromatogram evalu time depend correspond abovement criterion polydispers depend molecular weight standard case polystyren standard typic valu gt tabl gt descript test method prepar standard polystyren solut polystyren standard dissolv care mix chosen eluent recommend manufactur account prepar solut concentr standard chosen depend factor inject volum viscos solut sensit analyt detector maximum inject volum adapt length column order avoid overload typic inject volum analyt separ gpc column cm mm ìl higher volum exce ìl optim ratio inject volum concentr determin prior actual calibr column prepar sampl solut principl requir appli prepar sampl solut sampl dissolv suitabl solvent tetrahydrofuran thf shake carefulli circumst dissolv ultrason bath sampl solut purifi membran filter pore size ìm presenc undissolv particl record final report due high molecular weight speci method determin percentag weight dissolv particl solut hour apparatu solvent reservoir degass pump puls dampen inject system chromatographi column detector flowmet data record processor wast vessel ensur gpc system inert regard utilis solvent steel capillari thf solvent inject solvent deliveri system defin volum sampl solut load column auto sampler manual sharpli defin zone withdraw depress plunger syring quickli manual observ molecular weight distribut solvent deliveri system pulsat free ideal incorpor puls dampen flow rate order ml min column depend sampl polym characteris simpl column column connect sequenc number porou column materi defin properti pore size exclus limit commerci select separ gel length column depend properti sampl hydrodynam volum molecular weight distribut specif condit separ solvent temperatur flow rate theoret plate column combin column separ characteris number theoret plate involv case thf elut solvent load solut ethyl benzen suitabl polar solut column length number theoret plate equat gt num gt ve gt den gt gt num gt ve gt den gt number theoret plate ve elut volum peak maximum baselin peak width peak width half height separ effici addit number theoret plate quantiti determin bandwidth part plai separ effici determin steep calibr curv separ effici column obtain relationship gt num gt ve ve gt den gt cross section area column amp ge gt num gt cm gt den gt cm ve elut volum polystyren molecular weight mx ve elut volum polystyren time greater molecular weight resolut system commonli defin gt num gt ve ve gt den gt gt num gt gt den gt log ve ve elut volum polystyren standard peak maximum peak width baselin molecular weight peak maximum differ factor column system greater solvent solvent high puriti thf puriti solvent reservoir inert ga atmospher suffici larg calibr column sampl analys solvent degass transport column pump temperatur control temperatur critic intern compon inject loop column detector tube constant consist choic solvent detector purpos detector record quantit concentr sampl elut column order avoid unnecessari broaden peak cuvett volum detector cell small larger ìl light scatter viscos detector differenti refractometri detect requir specif properti sampl elut solvent type detector uv vi ir viscos detector data report data din standard refer detail evalu criteria requir relat collect process data sampl independ experi carri analys individu mn mw mw mn mp provid measur explicitli measur valu rel valu equival molecular weight standard determin retent volum retent time possibl correct intern standard log mp valu mp peak maxima calibr standard plot quantiti calibr point molecular weight decad measur point requir total curv cover estim molecular weight sampl low molecular weight end point calibr curv defin hexyl benzen suitabl polar solut number averag weight averag molecular weight gener determin mean electron data process base formula section case manual digitis astm consult distribut curv provid form tabl figur differenti frequenc sum percentag log graphic represent molecular weight decad cm width peak maximum cm height case integr distribut curv differ ordin cm test report test report includ inform test substanc inform test substanc ident addit impur descript treatment sampl observ problem instrument reservoir eluent inert ga degass eluent composit eluent impur pump puls dampen inject system separ column manufactur inform characterist column pore size kind separ materi number length order column number theoret plate column combin separ effici resolut system inform symmetri peak column temperatur kind temperatur control detector measur principl type cuvett volum flowmet manufactur measur principl system record process data hardwar softwar calibr system detail descript method construct calibr curv inform qualiti criteria method correl coeffici error sum squar inform extrapol assumpt approxim made experiment procedur evalu process data measur construct calibr curv document tabl includ inform calibr point sampl manufactur sampl characterist valu standard mp mn mw mw mn provid manufactur deriv subsequ measur detail method determin inject volum inject concentr mp calibr elut volum correct retent time measur peak maxima mp calcul peak maximum percentag error calcul mp calibr evalu evalu time basi method ensur requir reproduc method correct intern standard inform evalu effect basi elut volum retent time inform limit evalu peak complet analys descript smooth method prepar pre treatment procedur sampl presenc undissolv particl inject volum ìl inject concentr mg ml observ indic effect lead deviat ideal gpc profil detail descript modif test procedur detail error rang inform observ relev interpret result refer din gelpermeationschromatographi gpc mit tetrahydrofuran thf al elutionsmittel teil yau kirkland bly ed modern size exclus liquid chromatographi wilei son astm standard test method molecular weight averag molecular weight distribut liquid exclus chromatographi gel permeat chromatographi gpc american societi test materi philadelphia pennsylvania astm standard test method molecular weight averag molecular weight distribut polystyren high perform size exclus chromatographi american societi test materi philadelphia pennsylvania annex exampl method determin number averag molecular weight mn polym gel permeat chromatographi gpc prefer method determin mn set standard structur compar polym structur practic difficulti gpc expect substanc fail regulatori mn criterion confirm altern method collig properti ebullioscopi cryoscopi involv measur boil point elev ebullioscopi freez point depress cryoscopi solvent polym ad method reli fact effect dissolv polym boil freez point liquid depend molecular weight polym applic mn annex iii low molecular weight content polym method gel permeat chromatograph method replic oecd tg fundament principl technic inform refer introduct properti polym vari imposs describ singl method set precis condit separ evalu cover eventu specif occur separ polym complex polym system amen gel permeat chromatographi gpc gpc practic molecular weight determin mean method annex case full detail justif method method base din standard detail inform carri experi evalu data found din standard case modif experiment condit justifi standard fulli referenc method polystyren sampl polydispers calibr modifi suitabl polym water solubl long chain branch polym definit unit low molecular weight arbitrarili defin molecular weight dalton number averag molecular weight mn weight averag molecular weight mw determin equat mn gt num gt lnhi gt den gt ln gt num gt gt den gt mi mw gt num gt lnhixmi gt den gt lnhi level detector signal baselin retent volum vi mi molecular weight polym fraction retent volum vi number data point breadth molecular weight distribut measur dispers system ratio mw mn refer substanc gpc rel method calibr undertaken narrowli distribut linearli construct polystyren standard averag molecular weight mn mw molecular weight distribut calibr curv determin molecular weight unknown sampl condit separ sampl standard select ident manner determin relationship molecular weight elut volum valid specif condit experi condit includ temperatur solvent solvent mixtur chromatographi condit separ column system column molecular weight sampl determin rel valu polystyren equival molecular weight mean depend structur chemic differ sampl standard molecular weight deviat absolut valu greater lesser degre standard polyethylen glycol polyethylen oxid polymethyl methacryl polyacryl acid reason state principl test method molecular weight distribut sampl averag molecular weight mn mw determin gpc gpc special type liquid chromatographi sampl separ hydrodynam volum individu constitu separ effect sampl pass column fill porou materi typic organ gel small molecul penetr pore larg molecul exclud path larg molecul shorter elut medium size molecul penetr pore elut smallest molecul hydrodynam radiu smaller pore gel penetr pore elut ideal situat separ govern size molecular speci practic difficult avoid absorpt effect interf uneven column pack dead volum worsen situat detect effect refract index uv absorpt yield simpl distribut curv attribut actual molecular weight valu curv calibr column pass polym molecular weight ideal broadli similar structur polystyren standard typic gaussian curv result distort small tail low molecular weight side vertic axi indic quantiti weight molecular weight speci elut horizont axi log molecular weight low molecular weight content deriv curv calcul accur low molecular weight speci respond equival mass basi polym qualiti criteria repeat rel standard deviat rsd elut volum requir repeat analysi ensur correct intern standard chromatogram evalu time depend correspond abovement criterion polydispers depend molecular weight standard case polystyren standard typic valu gt tabl gt descript test method prepar standard polystyren solut polystyren standard dissolv care mix chosen eluent recommend manufactur account prepar solut concentr standard chosen depend factor inject volum viscos solut sensit analyt detector maximum inject volum adapt length column order avoid overload typic inject volum analyt separ gpc column cm mm ìl higher volum exce ìl optim ratio inject volum concentr determin prior actual calibr column prepar sampl solut principl requir appli prepar sampl solut sampl dissolv suitabl solvent tetrahydrofuran thf shake carefulli circumst dissolv ultrason bath sampl solut purifi membran filter pore size ìm presenc undissolv particl record final report due high molecular weight speci method determin percentag weight undissolv particl solut hour correct content impur addit correct content speci num gt ve gt den gt gt num gt ve gt den gt number theoret plate ve elut volum peak maximum baselin peak width peak width half height separ effici addit number theoret plate quantiti determin bandwidth part plai separ effici determin steep calibr curv separ effici column obtain relationship gt num gt ve ve gt den gt cross section area column amp ge gt num gt cm gt den gt cm ve elut volum polystyren molecular weight mx ve elut volum polystyren time greater molecular weight resolut system commonli defin gt num gt ve ve gt den gt gt num gt gt den gt log ve ve elut volum polystyren standard peak maximum peak width baselin molecular weight peak maximum differ factor column system greater solvent solvent high puriti thf puriti solvent reservoir inert ga atmospher suffici larg calibr column sampl analys solvent degass transport column pump temperatur control temperatur critic intern compon inject loop column detector tube constant consist choic solvent detector purpos detector record quantit concentr sampl elut column order avoid unnecessari broaden peak cuvett volum detector cell small larger ìl light scatter viscos detector differenti refractometri detect requir specif properti sampl elut solvent type detector uv vi ir viscos detector data report data din standard refer detail evalu criteria requir relat collect process data sampl independ experi carri analys individu case essenti determin data blank treat condit sampl explicitli measur valu rel valu equival molecular weight standard determin retent volum retent time possibl correct intern standard log mp valu mp peak maxima calibr standard plot quantiti calibr point molecular weight decad measur point requir total curv cover estim molecular weight sampl low molecular weight end point calibr curv defin hexyl benzen suitabl polar solut portion curv molecular weight determin correct impur addit elut curv gener evalu mean electron data process case manual digitis astm consult insolubl polym retain column molecular weight higher solubl fraction consid result overestim low molecular weight content guidanc correct low molecular weight content insolubl polym provid annex distribut curv provid form tabl figur differenti frequenc sum percentag log graphic represent molecular weight decad cm width peak maximum cm height case integr distribut curv differ ordin cm test report test report includ inform test substanc inform test substanc ident addit impur descript treatment sampl observ problem instrument reservoir eluent inert ga degass eluent composit eluent impur pump puls dampen inject system separ column manufactur inform characterist column pore size kind separ materi number length order column number theoret plate column combin separ effici resolut system inform symmetri peak column temperatur kind temperatur control detector measur principl type cuvett volum flowmet manufactur measur principl system record process data hardwar softwar calibr system detail descript method construct calibr curv inform qualiti criteria method correl coeffici error sum squar inform extrapol assumpt approxim made experiment procedur evalu process data measur construct calibr curv document tabl includ inform calibr point sampl manufactur sampl characterist valu standard mp mn mw mw mn provid manufactur deriv subsequ measur detail method determin inject volum inject concentr mp calibr elut volum correct retent time measur peak maxima mp calcul peak maximum percentag error calcul mp calibr inform low molecular weight polym content descript method analysi experi conduct inform percentag low molecular weight speci content relat total sampl inform impur addit polym speci percentag weight relat total sampl evalu evalu time basi method ensur requir reproduc method correct intern standard inform evalu effect basi elut volum retent time inform limit evalu peak complet analys descript smooth method prepar pre treatment procedur sampl presenc undissolv particl inject volum ìl inject concentr mg ml observ indic effect lead deviat ideal gpc profil detail descript modif test procedur detail error rang inform observ relev interpret result refer din gelpermeationschromatographi gpc mit tetrahydrofuran thf al elutionsmittel teil yau kirkland bly ed modern size exclus liquid chromatographi wilei son astm standard test method molecular weight averag molecular weight distribut liquid exclus chromatographi gel permeat chromatographi gpc american societi test materi philadelphia pennsylvania astm standard test method molecular weight averag molecular weight distribut polystyren high perform size exclus chromatographi american societi test materi philadelphia pennsylvania annex guidanc correct low molecular content presenc insolubl polym insolubl polym present sampl result mass loss gpc analysi insolubl polym irrevers retain column sampl filter solubl portion sampl pass column case refract index increment dn dc polym estim measur estim sampl mass lost column case make correct extern calibr standard materi concentr dn dc calibr respons refractomet poli methyl methacryl pmma standard extern calibr analysi acryl polym pmma standard concentr tetrahydrofuran analys gpc result data find refractomet constant equat gt num gt gt den gt gt num gt dn gt den gt dc refractomet constant microvoltsecond ml respons pmma standard microvoltsecond concentr pmma standard mg ml inject volum ml dn dc refract index increment pmma tetrahydrofuran ml mg data typic pmma standard mg ml ml dn dc ml mg result calcul theoret detector respons polym inject elut detector annex iii solut extract behaviour polym water method method replic revis version oecd tg technic inform refer introduct polym emuls polym initi preparatori work method set method applic liquid polym polym react water test condit method practic solut extract behaviour investig mean method case full detail justif method refer substanc principl test method solut extract behaviour polym aqueou medium determin flask method water solubl flask method modif qualiti criteria descript test method equip equip requir method crush devic grinder product particl size apparatu shake possibl temperatur control membran filter system analyt equip standardis siev sampl prepar repres sampl reduc particl size mm siev cool requir stabil sampl grind process materi rubberi natur crush liquid nitrogen temperatur requir particl size fraction attain action reduc particl size result report report crush sampl store prior test procedur sampl test substanc weigh vessel fit glass stopper ml water ad vessel handl amount polym prove impractic highest amount handl volum water adjust vessel tightli stopper agit shake stir devic capabl oper constant temperatur period hour content vessel centrifug filter concentr polym clear aqueou phase determin suitabl analyt method suitabl analyt method aqueou phase total solubl extract estim dry weight filter residu centrifug precipit differenti quantit impur addit hand low molecular weight speci hand case gravimetr determin import perform blank run test substanc order account residu aris experiment procedur solut extract behaviour polym water ph ph determin conduct experi ph valu achiev addit suitabl buffer acid base hydrochlor acid acet acid analyt grade sodium potassium hydroxid nh depend method analysi test perform suffici specif method direct analysi aqueou phase polym compon test suffic method determin solut extract behaviour polym limit indirect analysi determin total organ carbon content toc aqueou extract addit test conduct addit test triplic time smaller polym sampl amount water test analysi test conduct sampl size method direct analysi polym compon aqueou phase altern indirect analysi dissolv extract polym compon determin total content solubl part correct polym specif compon consid analysi aqueou phase total polymer speci suffici sensit method toc persulph dichrom digest yield estim ir chemic analysi atom absorpt spectrometri aa induct coupl plasma icp emiss equival silicon metal polym uv absorpt spectrofluorimetri aryl polym lc ms low molecular weight sampl vacuum evapor dryness aqueou extract spectroscop ir uv aa icp analysi residu analysi aqueou phase practic aqueou extract extract water immisc organ solvent chlorin hydrocarbon solvent evapor residu analys notifi polym content compon residu identifi impur addit subtract purpos determin degre solut extract polym larg quantiti materi present subject residu hplc gc analysi differenti impur monom monom deriv speci present true content determin case simpl evapor organ solvent dryness weigh dry residu suffici test conduct sampl size aqueou extract analys toc gravimetr determin perform undissolv extract part sampl centrifug filter content vessel polym residu remain attach wall vessel vessel rins filtrat vessel clear visibl residu filtrat centrifug filter residu remain filter centrifug tube dri vacuum weigh dry continu constant weight reach data test conduct sampl size individu result flask averag valu express unit mass volum solut typic mg mass mass polym sampl typic mg addition weight loss sampl calcul weight solut divid weight initi sampl rel standard deviat rsd calcul individu figur total substanc polym essenti addit polym subtract contribut addit test conduct sampl size individu toc valu aqueou extract triplic experi averag experi express unit mass volum solut typic mgc unit mass weight initi sampl typic mgc differ result high low sampl water ratio extract compon extract case direct analysi individu weight residu express percentag initi weight sampl averag calcul experi differ percentag found repres percentag solubl extract materi origin sampl report test report test report includ inform test substanc inform test substanc ident addit impur content low molecular weight speci experiment condit descript procedur experiment condit descript analyt detect method result result solubl extract mg individu valu extract test solut broken polym content impur addit result solubl extract mg polym toc valu aqueou extract weight solut calcul percentag measur ph sampl inform blank valu refer chemic instabl test substanc test process analyt process inform import interpret result refer din zerkleinerung von kunststofferzeugnissen für prüfzweck annex iii bioconcentr flow fish test method bioconcentr method replic oecd tg introduct method describ procedur characteris bioconcentr potenti substanc fish flow condit flow test regim prefer semi static regim permiss provid valid criteria satisfi method suffici detail perform test allow adequ freedom adapt experiment design condit laboratori vari characterist test substanc validli appli stabl organ chemic log pow valu appli superlipophil substanc log pow gt pre estim bioconcentr factor bcf denot kb superlipophil substanc higher steadi state bioconcentr factor bcfss expect obtain laboratori experi pre estim bioconcentr factor organ chemic log pow valu obtain equat bintein al paramet characteris bioconcentr potenti includ uptak rate constant depur rate constant bcfss radio label test substanc facilit analysi water fish sampl determin degrad identif quantif made total radioact residu measur combust tissu solubilis bcf base parent compound retain metabolit assimil carbon bcf base total radioact residu directli compar bcf deriv specif chemic analysi parent compound clean procedur emploi radio label studi order determin bcf base parent compound major metabolit characteris deem combin fish metabol studi bioconcentr studi analysi identif residu tissu definit unit bioconcentr bioaccumul increas concentr test substanc organ tissu thereof rel concentr test substanc surround medium bioconcentr factor bcf kb time uptak phase accumul test concentr test substanc fish tissu thereof cf ìg ppm divid concentr chemic surround medium cw ìg ml ppm steadi state bioconcentr factor bcfss kb chang significantli prolong period time concentr test substanc surround medium constant period time plateau steadi state reach plot test substanc fish cf time curv parallel time axi success analys cf made sampl interv dai signific differ sampl period pool sampl analys success analys requir test substanc slowli interv appropri dai bioconcentr factor calcul directli kinet rate constant term kinet concentr factor bcfk octanol water partit coeffici pow ratio chemic solubl octanol water equilibrium method express kow logarithm pow indic chemic potenti bioconcentr aquat organ exposur uptak phase time fish expos test chemic uptak rate constant numer defin rate increas concentr test substanc test fish tissu thereof fish expos chemic express dai post exposur depur loss phase time transfer test fish medium test substanc medium free substanc depur net loss substanc test fish tissu thereof studi depur loss rate constant numer defin rate reduct concentr test substanc test fish tissu thereof transfer test fish medium test substanc medium free substanc express dai principl test method test consist phase exposur uptak post exposur depur phase uptak phase separ group fish speci expos concentr test substanc transfer medium free test substanc depur phase depur phase uptak substanc uptak phase insignific bcf concentr test substanc fish tissu thereof phase test addit test concentr control group fish held ident condit absenc test substanc relat advers effect observ bioconcentr test match control group obtain background concentr test substanc uptak phase run dai demonstr equilibrium reach earlier predict length uptak phase time steadi state made equat annex depur period begun transfer fish medium test substanc clean vessel bioconcentr factor calcul prefer ratio bcfss concentr fish cf water cw appar steadi state kinet bioconcentr factor bcfk ratio rate constant uptak depur assum order kinet order kinet obei complex model emploi annex steadi state achiev dai uptak phase extend steadi state reach dai whichev depur phase begun uptak rate constant depur loss rate constant constant complex model involv bioconcentr factor confid limit paramet calcul model describ measur concentr test substanc fish water bcf express function total wet weight fish special purpos tissu organ muscl liver fish suffici larg fish divid edibl fillet edibl viscera fraction organ substanc clear relationship potenti bioconcentr lipophil relationship lipid content test fish observ bioconcentr substanc reduc sourc variabl test result substanc high lipophil log pow gt bioconcentr express relat lipid content addit bodi weight lipid content determin biolog materi determin concentr test substanc feasibl inform test substanc carri test bioconcentr inform test substanc solubl water octanol water partit coeffici pow denot kow determin hplc method hydrolysi phototransform water determin solar simul solar irradi irradi condit test bioconcentr surfac tension substanc log pow determin vapour pressur readi biodegrad inform requir toxic fish speci test prefer asymptot lc time independ analyt method accuraci precis sensit quantif test substanc test solut biolog materi detail sampl prepar storag analyt detect limit test substanc water fish tissu label test substanc percentag radioact impur valid test condit appli test valid temperatur variat concentr dissolv oxygen fall satur concentr test substanc chamber maintain measur valu uptak phase mortal advers effect diseas control treat fish end test test extend week month death advers effect set fish month exce refer compound refer compound bioconcentr potenti check experiment procedur requir specif substanc recommend descript test method apparatu care avoid materi part equip dissolv sorb leach advers effect fish standard rectangular cylindr tank made chemic inert materi suitabl capac complianc load rate soft plastic tube minimis teflon stainless steel glass tube prefer experi shown substanc high absorpt coeffici synthet pyrethroid silanis glass requir situat equip discard water natur water gener test obtain uncontamin uniform qualiti sourc dilut water qualiti surviv chosen fish speci durat acclim test period show abnorm appear behaviour ideal demonstr test speci surviv grow reproduc dilut water laboratori cultur life cycl toxic test water characteris ph hard total solid total organ carbon prefer ammonium nitrit alkalin marin speci salin paramet import optim fish fulli annex recommend maximum concentr number paramt fresh marin test water water constant qualiti period test ph rang test rang ph unit order ensur dilut water unduli influenc test result complex test substanc advers affect perform stock fish sampl interv analysi determin heavi metal cu pb zn hg cd ni major anion cation ca mg na cl pesticid total organophosphor total organochlorin pesticid total organ carbon suspend solid made month dilut water constant qualiti water qualiti demonstr constant year determin frequent interv extend month natur particl content total organ carbon toc dilut water low avoid absorpt test substanc organ matter reduc bioavail maximum accept mg particul matter dry matter pass ìm filter mg total organ carbon annex water filter contribut organ carbon content water test fish excreta food residu low test concentr organ carbon test vessel exce concentr organ carbon origin test substanc solubilis agent mg test solut stock solut test substanc prepar suitabl concentr stock solut prefer prepar simpli mix agit test substanc dilut water solvent dispers solubilis agent recommend occur case order produc suitabl concentr stock solut solvent ethanol methanol ethylen glycol monomethyl ether etheylen glycol dimethyl ether dimethylformamid triethylen glycol dispers cremophor rh tween methylcellulos hco care readili biodegrad agent problem bacteri growth flow test test substanc radio label highest puriti prefer gt flow test system continu dispens dilut stock solut test substanc meter pump proport dilut satur system requir deliv test concentr test chamber volum replac test chamber dai prefer allow flow mode prefer test organ advers affect semi static techniqu provid valid criteria satisfi flow rate stock solut dilut water check hour daili test check determin flow rate test chamber includ ensur vari chamber select speci import criteria select speci readili obtain conveni size satisfactorili maintain laboratori criteria select fish speci includ recreat commerci ecolog import compar sensit past success recommend test speci annex speci test procedur adapt provid suitabl test condit rational select speci experiment method report case hold fish acclim stock popul fish week water test temperatur feed suffici diet type test hour settl period mortal record criteria appli mortal greater popul dai reject entir batch mortal popul dai acclim addit dai mortal popul dai accept batch mortal dai reject entir batch ensur fish test free observ diseas abnorm discard diseas fish fish receiv treatment diseas week preced test test perform test preliminari test conduct preliminari experi order optimis test condit definit test select test substanc concentr durat uptak depur phase condit exposur durat uptak phase predict durat uptak phase obtain practic experi previou studi accumul relat chemic empir relationship utilis knowledg aqueou solubl octanol water partit coeffici test substanc annex uptak phase run dai demonstr equilibrium reach earlier steadi state reach dai uptak phase extend take measur steadi state reach dai whichev shorter durat depur phase period half durat uptak phase suffici reduct bodi burden substanc occur annex explan estim time requir reach loss impract long exceed normal durat uptak phase dai shorter period concentr test substanc steadi state concentr substanc complex pattern uptak depur repres compart fish model yield order kinet longer depur phase determin loss rate constant period govern period concentr test substanc fish remain analyt detect limit number test fish select number fish test concentr minimum fish sampl sampl greater statist power requir fish sampl adult fish report male femal experi sex differ lipid content sex document signific start exposur pool male femal fish test fish similar weight select smallest smaller third weight largest year class sourc weight ag fish signific effect bcf valu detail record accur recommend sampl stock fish weigh test order estim weight load high water fish ratio order minimis reduct cw caus addit fish start test avoid decreas dissolv oxygen concentr import load rate test speci case load rate fish wet weight litr water dai recommend high load rate shown requir concentr test substanc maintain limit concentr dissolv oxygen fall satur choos load regim account normal habitat fish speci bottom live fish demand larger bottom area aquarium volum water pelag fish speci feed acclim test period fish fed diet lipid total protein content amount suffici healthi condit maintain bodi weight fish fed daili acclim test period level approxim bodi weight dai lipid concentr speci fish constant level test amount feed calcul week order maintain consist bodi weight lipid content calcul weight fish test chamber estim weight fish sampl recent chamber weigh fish remain chamber uneaten food faec siphon daili test chamber shortli feed minut hour chamber clean test concentr organ matter low presenc organ carbon limit bioavail test substanc feed deriv fishmeal feed analys test substanc desir analys feed pesticid heavi metal light temperatur photoperiod hour temperatur test speci annex type characterist illumin caution phototransform test substanc irradi condit studi illumin avoid exposur fish unnatur photoproduct case filter screen uv irradi nm test concentr fish expos flow condit concentr test substanc water higher highest concentr test substanc select acut asymptot lc ten fold higher detect limit water analyt method highest test concentr determin divid acut lc acut chronic ratio ratio chemic choos concentr differ factor lc criterion analyt limit lower factor label test substanc consid concentr solubl test substanc solubilis agent concentr greater ml test vessel contribut test substanc content organ carbon test water effort made avoid materi control dilut water control relev control solubilis agent run addit test seri provid establish agent effect fish control set frequenc water qualiti measur test dissolv oxygen toc ph temperatur measur vessel total hard salin relev measur control vessel higher highest concentr minimum dissolv oxygen salin relev measur time begin middl end uptak period week depur period toc measur begin test hour hour prior test initi uptak phase addit fish week uptak depur phase temperatur measur daili ph begin end period hard test temperatur prefer monitor continu vessel sampl analysi fish water fish water sampl schedul water test chamber determin test substanc concentr sampl addit fish uptak depur phase minimum water sampl time fish feed uptak phase concentr test substanc determin order check complianc valid criteria fish sampl occas uptak phase occas depur phase occas difficult calcul precis estim bcf base number sampl simpl order depur kinet advis sampl higher frequenc period annex extra sampl store analys result round analys prove inadequ calcul bcf desir precis accept sampl schedul annex schedul readili calcul assum valu pow calcul exposur time uptak sampl continu uptak phase steadi state establish dai whichev shorter steadi state reach dai sampl continu steadi state attain dai whichev shorter begin depur phase fish transfer clean tank sampl sampl prepar water sampl analysi obtain siphon inert tube central point test chamber filtrat centrifug separ bioavail fraction test substanc bioavail super lipophil chemic chemic log pow gt sampl subject treatment measur tank clean content total organ carbon monitor uptak depur phase number fish minimum remov test chamber sampl time sampl fish rins quickli water blot dry kill instantli human method weigh prefer analys fish water immedi sampl order prevent degrad loss calcul approxim uptak depur rate test proce analysi avoid delai determin plateau reach fail analysi sampl store method begin studi inform proper method storag test substanc deep freez hold durat storag extract obtain qualiti analyt method procedur govern essenti accuraci precis sensit analyt method test substanc check experiment precis reproduc chemic analysi recoveri test substanc water fish satisfactori method check test substanc detect dilut water valu cw cf obtain test correct recoveri background valu control fish water sampl handl manner minimis contamin loss result adsorpt sampl devic analysi fish sampl radiolabel materi test analys total radio label parent metabolit sampl clean parent compound analys separ major metabolit characteris steadi state end uptak phase whichev sooner bcf term total radiolabel residu amp ge advis categori chemic pesticid strongli recommend identifi quantifi degrad repres amp ge total residu fish tissu steadi state degrad repres amp ge total radiolabel residu fish tissu identifi quantifi recommend identifi quantifi degrad test water concentr test substanc determin weigh individu fish pool sampl sampl occas pool restrict statist procedur appli data specif statist procedur power import consider adequ number fish accommod desir pool procedur power includ test bcf express function total wet weight high lipophil substanc function lipid content lipid content fish determin sampl occas suitabl method determin lipid content refer annex chloroform methanol extract techniqu recommend standard method method give ident valu import give detail method analysi lipid made extract produc analysi test substanc lipid remov extract analys chromatograph lipid content fish mg kg wet weight end experi differ start tissu percent solid report convers lipid concentr wet dry basi data treatment result uptak curv test substanc obtain plot concentr fish tissu uptak phase time arithmet scale curv reach plateau approxim asymptot time axi steadi state bcfss calcul gt num gt cf steadi state gt den gt cw steadi state steadi state reach calcul bcfss suffici precis hazard assess steadi state equilibrium concentr factor bcfk determin ratio order kinet constant depur rate constant determin depur curv plot decreas test substanc concentr fish time uptak rate constant calcul cf deriv uptak curv annex prefer method obtain bcfk rate constant linear paramet estim method comput graphic method calcul depur curv order complex model emploi refer annex advic biostatistician sought interpret result result interpret caution measur concentr test solut occur level detect limit analyt method defin uptak loss curv indic good qualiti bioconcentr data variat uptak depur constant test concentr observ signific differ uptak depur rate appli test concentr record explan gener confid limit bcf design studi approach report test report includ inform test substanc physic natur relev physicochem properti chemic identif data includ organ carbon content radio label precis posit label atom percentag radioact impur test speci scientif strain sourc pre treatment acclim ag size rang test condit test procedur flow semi static type characterist illumin photoperiod test design number size test chamber water volum replac rate number replic number fish replic number test concentr length uptak depur phase sampl frequenc fish water sampl method prepar stock solut frequenc renew solubilis agent concentr contribut organ carbon content test water nomin test concentr mean measur valu standard deviat test vessel method attain sourc dilut water descript pre treatment result demonstr abil test fish live water water characterist ph hard temperatur dissolv oxygen concentr residu chlorin level measur total organ carbon suspend solid salin test medium measur made water qualiti test vessel ph hard toc temperatur dissolv oxygen concentr detail inform feed type food sourc composit lipid protein content amount frequenc inform treatment fish water sampl includ detail prepar storag extract analyt procedur precis test substanc lipid content measur result result preliminari studi perform mortal control fish fish exposur chamber observ abnorm behaviour lipid content fish determin test occas curv includ measur data show uptak depur test chemic fish time steadi state cf cw standard deviat rang sampl time cf express ìg wet weight ppm bodi tissu thereof lipid cw ìg ml ppm cw valu control seri background report steadi state bioconcentr factor bcfss kinet concentr factor bcfk applic confid limit uptak depur loss rate constant express relat bodi total lipid content measur anim tissu thereof confid limit standard deviat method comput data analysi concentr test substanc radio label substanc requir accumul detect metabolit present unusu test deviat procedur relev inform minimis result detect limit detect pre test method develop experiment design result rate constant calcul refer connel bioaccumul behaviour persist chemic aquat organ rev environ contam toxicol pp bintein devil karcher linear depend fish bioconcentr octanol water partit coeffici sar qsar environment research pp oecd pari direct phototransform chemic water environment health safeti guidanc document seri test assess chemic kristensen bioconcentr fish comparison bioconcentr factor deriv oecd astm test method influenc particul organ matter bioavail chemic water qualiti institut denmark epa great lake water qualiti initi technic support document procedur determin bioaccumul factor juli fda food drug administr revis pesticid analyt manual fisher lane rockvil md juli epa section analysi human anim adipos tissu analysi pesticid residu human environment sampl thompson ed research triangl park compaan determin effect chemic wast aquat environ degrad toxic bioaccumul ch part ii govern publish offic hagu netherland gardner al limn amp oceanogr pp randal lee ozretich lake pruell evalu select lipid method normalis pollut bioaccumul envir toxicol chem pp cec bioconcentr chemic substanc fish flow method ring test programm final report march author kristensen nyholm astm reapprov standard practic conduct bioconcentr test fish saltwat bivalv mollusc annex gt tabl gt annex gt tabl gt estuarin marin speci countri gt tabl gt collect fresh water fish list tabl easi rear wide year avail marin estuarin speci partial confin respect countri capabl bred cultiv fish farm laboratori diseas parasit control condit test anim healthi parentag fish part world annex predict durat uptak depur phase predict durat uptak phase perform test estim percentag time need reach steadi state obtain empir relationship octanol water partit coeffici pow aqueou solubl estim dai obtain empir relationship log log pow equat relationship refer partit coeffici pow estim made knowledg aqueou solubl substanc log pow log equat solubl mole relationship appli chemic log pow valu time reach percentag steadi state obtain appli estim gener kinet equat describ uptak depur order kinet gt num gt dcf gt den gt dt cw cf cw constant cf gt num gt gt den gt cw equat steadi state approach amp infin equat reduc cf gt num gt gt den gt cw cf cw bcf cw approach concentr fish steadi state cf equat transcrib cf cf gt num gt cf gt den gt cf equat appli equat time reach percentag steadi state predict pre estim equat guidelin statist optim durat uptak phase product statist accept data bcfk period requir curv logarithm concentr test substanc fish plot linear time reach mid point steadi state steadi state time reach steadi state equat gt num gt gt den gt equat similarli steadi state gt num gt gt den gt equat durat uptak phase test substanc log pow equat log dai dai hour orup dai hour similarli test substanc mol log durat equat log pow log dai dai hour orup dai hour altern express teq pow hour calcul time effect steadi state reach predict durat depur phase predict time need reduc bodi burden percentag initi concentr obtain gener equat describ uptak depur order kinet depur phase cw assum equat reduc gt num gt dcf gt den gt dt cf cf cf kwhere cf concentr start depur period depur reach time gt num gt cf gt den gt cf gt num gt gt den gt gt num gt gt den gt similarli depur reach gt num gt gt den gt uptak period loss depur phase depur phase approxim durat uptak phase import note estim base assumpt uptak depur pattern follow order kinet order kinet obei complex model emploi refer literatur annex spaci hamelink altern model describ bioconcentr organ fish environ toxicol chem pp kristensen bioconcentr fish comparison bcf deriv oecd astm test method influenc particul matter bioavail chemic danish water qualiti institut chiou schmed partit organ compound octanol water system environ sci technol pp hawker connel influenc partit coeffici lipophil compound bioconcentr kinet fish wat re pp branson blau alexand neeli transact american fisheri societi pp ernst accumul aquat organ apprais test predict environment behaviour chemic ed sheehman kort klein bourdeau part pp scope john wilei amp son reilli bajramov blau branson sauerhoff guidelin optim design experi estim paramet order kenet model chem eng pp könemann van leeuwen toxicokinet fish accumul elimin chlorobenzen guppi chemospher pp annex gt tabl gt annex model discrimin bioconcentr data assum simpl compart paramet model rectilinear curv approxim point concentr fish depur phase plot semi log paper point rectilinear curv complex model emploi spaci hamelink refer annex graphic method determin depur loss rate constant plot concentr test substanc found sampl fish sampl time semi log paper slope line gt num gt ln cf cf gt den gt gt refer film gt note deviat straight line complex depur pattern order kinet graphic method appli resolv type depur deviat order kinet graphic method determin uptak rate constant calcul gt num gt cfk gt den gt cw equat cf read midpoint smooth uptak curv produc data log concentr plot versu time arithmet scale comput method calcul uptak depur loss rate constant prefer mean obtain bioconcentr factor rate constant linear paramet estim method comput program find valu set sequenti time concentr data model cf cw gt num gt gt den gt num gt gt den gt