commiss direct ec april establish commun method analysi determin amprolium diclazuril carbadox feedingstuff amend direct eec eec repeal direct eec text eea relev commiss european commun regard treati establish european commun regard council direct eec juli introduct commun method sampl analysi offici control feedingstuff amend act access austria finland sweden articl thereof direct eec stipul offici control feedingstuff purpos check complianc requir aris law regul administr provis govern qualiti composit carri commun method sampl analysi council direct eec novemb addit feedingstuff amend commiss regul stipul amprolium diclazuril content label substanc ad premixtur feedingstuff authoris carbadox feed addit withdrawn commiss regul decemb amend council direct eec addit feedingstuff withdraw authoris growth promot offici control illeg prohibit substanc commun method analysi establish check substanc commiss direct eec june establish commun method analysi offici control feedingstuff amend direct ec set method analysi inter alia determin mustard oil asnd theobromin method longer valid light advanc scientif technic knowledg intend purpos delet method fourth commiss direct eec decemb establish commun method analysi offici control feedingstuff amend direct ec set method analysi inter alia determin retinol vitamin method longer valid light advanc scientif technic knowledg intend purpos delet method retinol commiss direct eec march establish commun method analysi offici control feedingstuff amend direct eec set method analysi determin starch starch degrad product high molecular weight feedingstuff beet cossett beet pulp dri beet top leav potato pulp dri yeast product rich inulin greav amprolium ethopab dinitolmid nicarbazin menadion vitamin method direct longer valid light advanc scientif technic knowledg intend purpos repeal direct measur provid direct accord opinion stand committe feedingstuff adopt direct articl member state provid analys conduct view offici control amprolium diclazuril carbadox content feedginstuff premixtur carri method set annex hereto articl direct eec amend articl word quot mustard oil quot quot theobromin quot delet point annex delet articl direct eec amend articl delet annex ii delet articl direct eec repeal articl member state bring forc octob law regul administr provis compli provis direct immedi inform commiss thereof appli measur novemb member state adopt measur refer direct accompani refer time offici public procedur refer adopt member state articl direct enter forc dai public offici journal european commun articl direct address member state brussel april commiss franz fischler member commiss oj oj oj oj oj oj oj oj oj annex part determin amprolium amino propylpyrimidin yl methyl methyl pyridinium chlorid hydrochlorid purpos scope method determin amprolium feedingstuff premixtur detect limit mg kg limit determin mg kg principl sampl extract methanol water mixtur dilut mobil phase membran filtrat content amprolium determin cation exchang high perform liquid chromatographi hplc uv detector reagent methanol acetonitril hplc grade water hplc grade sodium dihydrogen phosphat solut mol dissolv sodium dihydrogen phosphat monohydr water ml graduat flask make mark water mix sodium perchlor solut mol dissolv sodium perchlor monohydr water ml graduat flask make mark water mix mobil phase hplc observ mixtur acetonitril sodium dihydrogen phosphat solut sodium perchlor solut prior filter μm membran filter dega solut ultrason bath minut standard substanc pure amprolium amino propylpyrimidin yl methyl mehyl pyridinium chlorid hydrochlorid amprolium stock standard solut μg ml weigh nearest mg mg amprolium ml graduat flask dissolv ml methanol place flask min ultrason bath ultrason treatment bring solut room temperatur make mark water mix temperatur amp lt solut stabl month amprolium intermedi standard solut ìg ml pipett ml stock standard solut ml graduat flask make mark extract solvent mix temperatur amp lt solut stabl month calibr solut transfer ml intermedi standard solut seri ml graduat flask make mark mobil phase mix solut correspond ìg amprolium ml solut prepar freshli extract solvent methanol water mixtur apparatu hplc equip inject system suitabl inject volum ìl liquid chromatograph column mm mm cation exchang nucleosil sa ìm pack equival uv detector variabl wavelength adjust diod arrai detector membran filter ptfe materi ìm membran filter ìm ultrason bath mechan shaker magnet stirrer procedur gener blank feed perform recoveri test blank feed analys check amprolium interf substanc present blank feed similar type sampl amprolium interf substanc detect recoveri test recoveri test carri analys blank feed fortifi addit quantiti amprolium similar present sampl fortifi level mg kg transfer ml stock standard solut ml conic flask evapor solut approxim ml add blank feed mix leav minut mix time proceed extract step altern blank feed similar type sampl recoveri test perform mean standard addit method case sampl analys fortifi quantiti amprolium similar present sampl sampl analys unfortifi sampl recoveri calcul subtract extract premixtur content amp lt amprolium feedingstuff weigh nearest sampl depend amprolium content ml conic flask add ml extract solvent place flask ultrason bath leav minut remov flask ultrason bath shake hour shaker stir magnet stirrer dilut aliquot extract mobil phase amprolium content ìg ml mix observ filter ml dilut solut membran filter proce hplc determin premixtur content amp gt amprolium weigh nearest premixtur depend amprolium content ml conic flask add ml extract solvent place flask ultrason bath leav minut remov flask ultrason bath shake hour shaker stir magnet stirrer dilut aliquot extract mobil phase amprolium content μg ml mix filter ml dilut solut membran filter proce hplc determin hplc determin paramet condit offer guidanc condit provid give equival result gt tabl gt check stabil chromatograph system inject time calibr solut μg ml constant peak height retent time achiev calibr graph inject calibr solut time determin peak height area concentr plot calibr graph peak height area calibr solut ordin concentr μg ml abscissa sampl solut inject sampl extract time volum calibr solut determin peak height area amprolium peak calcul result height area amprolium peak sampl solut determin concentr sampl solut μg ml refer calibr graph amprolium content mg kg sampl formula gt refer graphic gt mg kg volum extract solvent ml ml amprolium concentr sampl extract μg ml dilut factor mass test portion valid result ident ident analyt confirm chromatographi diod arrai detector spectra sampl extract calibr solut μg ml compar chromatographi sampl extract fortifi addit amount calibr solut amount ad amprolium similar amount amprolium found sampl extract height amprolium peak enhanc take account amount ad dilut extract peak width half height origin width amprolium peak unfortifi sampl extract diod arrai detect result evalu criteria wavelength maximum absorpt sampl standard spectra record peak apex chromatogram margin determin resolv power detect system diod arrai detect typic nm nm sampl standard spectra record peak apex chromatogram part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb standard analyt nm spectra upslop apex downslop peak produc sampl extract part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb spectrum peak apex criteria met presenc analyt confirm repeat differ result parallel determin carri sampl exce rel higher amprolium content mg kg mg kg mg kg amprolium content mg kg mg kg rel higher amprolium content mg kg recoveri fortifi blank sampl recoveri result collabor studi collabor studi arrang poultri feed sampl miner feed sampl premix sampl analys result tabl gt tabl gt number laboratori number singl valu sr standard deviat repeat cvr coeffici variat repeat sr standard deviat reproduc cvr coeffici variat reproduc observ sampl thiamin thiamin peak chromatogram appear shortli amprolium peak method amprolium thiamin separ amprolium thiamin separ column method replac acetonitril portion mobil phase methanol british pharmacopoeia spectrum amprolium solut mol hydrochlor acid mol show maxima nm nm absorb amount nm nm extract dilut mobil phase retent time amprolium peak shift significantli due ionic strength part determin diclazuril chlorphenyl dichloro tetrahydro dioxo triazin yl phenyl acetonitril purpos scope method determin diclazuril feedingstuff premixtur limit detect mg kg limit determin mg kg principl addit intern standard sampl extract acidifi methanol feedingstuff aliquot extract purifi solid phase extract cartridg diclazuril elut cartridg mixtur acidifi methanol water evapor residu dissolv dmf water premixtur extract evapor residu dissolv dmf water content diclazuril determin ternari gradient revers phase high perform liquid chromatographi hplc uv detector reagent water hplc grade ammonium acet tetrabutylammonium hydrogen sulphat tbh acetonitril hplc grade methanol hplc grade dimethylformamid dmf hydrochlor acid ml standard substanc diclazuril ii chlorphenyl dichloro tetrahydro dioxo triazin yl phenyl acetonitril guarante puriti diclazuril stock standard solut μg ml weigh nearest mg mg diclazuril standard substanc ml graduat flask dissolv dmf make mark dmf mix wrap flask aluminium foil amber flask store refriger temperatur amp lt solut stabl month diclazuril standard solut ìg ml transfer ml stock standard solut ml graduat flask make mark dmf mix wrap flask aluminium foil amber flask store refriger temperatur amp lt solut stabl month intern standard substanc dichloro chlorophenyl dihydro dioxo triazin yl methylbenzen acetonitril intern stock standard solut ìg ml weigh nearest mg mg intern standard substanc ml graduat flask dissolv dmf make mark dmf mix wrap flask aluminium foil amber flask store refriger temperatur amp lt solut stabl month intern standard solut ìg ml transfer ml intern stock standard solut ml graduat flask make mark dmf mix wrap flask aluminium foil amber flask store refriger temperatur amp lt solut stabl month intern standard solut premixtur mg ml nomin content diclazuril premixtur mg kg weigh nearest mg mg intern standard substanc ml graduat flask dissolv dmf ultrason bath make mark dmf mix wrap flask aluminium foil amber flask store refriger temperatur amp lt solut stabl month calibr solut ìg ml pipet ml diclazuril standard solut ml intern standard solut ml graduat flask add ml dmf make mark water mix solut prepar freshli solid phase extract cartridg bond elut size cc sorbent mass mg extract solvent acidifi methanol pipet ml hydrochlor acid ml methanol mix mobil phase hplc eluent ammonium acet tetrabutylammonium hydrogen sulphat solut dissolv ammonium acet tbh ml water mix eluent acetonitril eluent methanol apparatu mechan shaker equip ternari gradient hplc liquid chromatograph column hypersil od ìm pack mm mm equival uv detector variabl wavelength adjust diod arrai detector vacuum rotari evapor membran filter ìm vacuum manifold ultrason bath procedur gener blank feed blank feed analys check diclazuril interf substanc present blank feed similar type sampl analysi diclazuril interf substanc detect recoveri test recoveri test carri analys blank feed fortifi addit quantiti diclazuril similar present sampl fortifi level mg kg add ml stock standard solut blank feed mix leav min mix time proceed altern blank feed similar type sampl recoveri test perform mean standard addit method case sampl analys fortifi quantiti diclazuril similar present sampl sampl analys unfortifi sampl recoveri calcul subtract extract feedingstuff weigh nearest approxim sampl transfer ml conic flask add ml intern standard solut ml extract solvent stopper flask shake mixtur shaker overnight settl minut transfer ml aliquot supernat suitabl glass contain dilut ml water transfer solut extract cartridg pass appli vacuum wash cartridg ml mixtur extract solvent water discard collect fraction elut compound ml mixtur extract solvent water evapor fraction reach dryness mean rotari evapor dissolv residu ml dmf add ml water mix filter membran filter proce hplc determin premixtur weigh nearest approxim sampl transfer ml conic flask add ml intern standard solut ml extract solvent stopper flask shake mixtur overnight shaker settl minut transfer aliquot ml nomin content diclazuril premix mg kg supernat round bottom flask suitabl size evapor reach dryness reduc pressur mean rotari evapor redissolv residu ml dmf add ml water mix proce hplc determin hplc determin paramet condit offer guidanc condit provid give equival result gt tabl gt check stabil chromatograph system inject time calibr solut μg ml constant peak height retent time achiev calibr solut inject μl calibr solut time determin peak height area diclazuril intern standard peak sampl solut inject μl sampl solut time determin peak height area diclazuril intern standard peak calcul result feed diclazuril content mg kg sampl formula gt refer graphic gt mg kg hd peak height area diclazuril sampl solut peak height area intern standard sampl solut hd peak height area diclazuril calibr solut peak height area intern standard calibr solut βd diclazuril concentr calibr solut μg ml mass test portion volum sampl extract ml premixtur diclazuril content mg kg sampl formula gt refer graphic gt mg kg hd peak height area diclazuril calibr solut peak height area intern standard calibr solut hd peak height area diclazuril sampl solut peak height area intern standard sampl solut βd diclazuril concentr calibr solut mass test portion volum sampl extract ml nomin content diclazuril mg kg premixtur valid result ident ident analyt confirm chromatographi diod arrai detector spectra sampl extract calibr solut compar chromatographi sampl extract fortifi addit amount calibr solut amount ad diclazuril similar amount diclazuril found sampl extract height diclazuril peak intern standard peak enhanc take account amount ad dilut extract peak width half height origin width diclazuril peak intern standard peak unfortifi sampl extract diod arrai detect result evalu criteria wavelength maximum absorpt sampl standard spectra record peak apex chromatogram margin determin resolv power detect system diod arrai detect typic nm nm sampl standard spectra record peak apex chromatogram part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb standard analyt nm spectra upslop apex downslop peak produc sampl extract part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb spectrum peak apex criteria met presenc analyt confirm repeat differ result parallel determin carri sampl exce rel higher diclazuril content mg kg mg kg mg kg diclazuril content mg kg mg kg rel higher diclazuril content mg kg recoveri fortifi blank sampl recoveri result collabor studi collabor studi arrang sampl analys laboratori sampl consist premixtur mix organ matrix inorgan matrix theoret content mg diclazuril kg mix feed poultri made produc nl theoret content mg diclazuril kg laboratori instruct analys sampl duplic detail inform collabor studi found journal aoac intern volum result tabl gt tabl gt number laboratori number singl valu sr standard deviat repeat cvr coeffici variat repeat sr standard deviat reproduc cvr coeffici variat reproduct observ diclazuril respons previous demonstr linear rang concentr measur part determin carbadox methyl quinoxalinylmethylen carbaz dioxid purpos scope method determin carbadox feedingstuff premixtur prepar detect limit mg kg limit determin mg kg principl sampl equilibr water extract methanol acetonitril feedingstuff aliquot portion filter extract subject clean aluminium oxid column premixtur prepar aliquot portion filter extract dilut concentr water methanol acetonitril content carbadox determin revers phase highperform liquid chromatographi hplc uv detector reagent methanol acetonitril hplc grade acet acid aluminium oxid neutral activ grade methanol acetonitril mix ml methanol ml acetonitril acet acid dilut ml acet acid ml water sodium acet ch coona water hplc grade acet buffer solut mol ph dissolv sodium acet ml water adjust ph acet acid transfer ml graduat flask make mark water mix mobil phase hplc mix ml acet buffer solut ml acetonitril filter μm filter dega solut ultrasonif minut standard substanc pure carbadox methyl quinoxalinylmethylen carbaz dioxid carbadox stock standard solut μg ml point procedur weigh nearest mg mg carbadox standard substanc ml graduat flask dissolv methanol acetonitril ultrasonif ultrason treatment bring solut room temperatur make mark methanol acetonitril mix wrap flask aluminium foil amber glasswar store refriger temperatur amp lt solut stabl month calibr solut transfer ml stock standard solut seri ml calibr flask add ml water make mark methanol acetonitril mix wrap flask aluminium foil solut correspond ìg ml carbadox calibr solut freshli prepar note determin carbadox feedingstuff mg kg calibr solut concentr ìg ml prepar water methanol acetonitril mixtur mix ml water ml mixtur methanol acetonitril apparatu laboratori shaker magnet stirrer glass fibr filter paper whatman gf equival glass column length mm intern diamet approxim mm sinter glass frit draw valv note glass column fit stopcock glass column taper end case small glass wool plug insert lower end tamp glass rod hplc equip inject system suitabl inject volum ìl liquid chromatograph column mm mm ìm pack equival uv detector variabl wavelength adjust diod arrai detector oper rang nm membran filter ìm membran filter ìm ultrason bath procedur note carbadox light sensit carri procedur subdu light amber glasswar glasswar wrap aluminium foil gener blank feed perform recoveri test blank feed analys check carbadox interf substanc present blank feed similar type sampl analysi carbadox interf substanc detect recoveri test recoveri test carri analys blank feed fortifi addit quantiti carbadox similar present sampl fortifi level mg kg transfer ml stock standard solut ml conic flask evapor solut approxim ml stream nitrogen add blank feed mix wait minut proceed extract step altern blank feed similar type sampl recoveri test perform mean standard addit method case sampl fortifi quantiti carbadox similar present sampl sampl analys unfortifi sampl recoveri calcul subtract extract feedingstuff weigh nearest approxim sampl transfer ml conic flask add ml water mix equilibr minut add ml methanol acetonitril stopper shake minut shaker stir magnet stirrer filter solut glass fibr filter paper retain solut purif step premixtur weigh nearest approxim unground sampl transfer ml conic flask add ml water mix equilibr minut add ml methanol acetonitril stopper shake minut shaker stir magnet stirrer filter solut glass fibr filter paper pipet aliquot filtrat ml calibr flask add ml water make mark methanol acetonitril mix carbadox concentr final solut approxim ìg ml aliquot filter ìm filter proce hplc determin prepar amp gt weigh nearest approxim unground sampl transfer ml conic flask add ml water mix equilibr minut add ml methanol acetonitril stopper homogenis sonic sampl minut shake stir minut filter solut glass fibr filter paper dilut aliquot filtrat mixtur water methanol acetonitril final carbadox concentr μg ml prepar dilut factor aliquot filter μm filter proce hplc determin purif prepar aluminium oxid column weigh aluminium oxid transfer glass column sampl purif appli ml filter extract aluminium oxid column discard ml eluat collect ml filter aliquot μm filter proce hplc determin hplc determin paramet condit offer guidanc condit provid yield equival result gt tabl gt check stabil chromatograph system inject calibr solut μg ml time constant peak height area retent time achiev calibr graph inject calibr solut time measur peak height area concentr plot calibr curv peak height area calibr solut ordin concentr μg ml abscissa sampl solut inject sampl extract feedingstuff premixtur prepar time determin peak height area carbadox peak calcul result height area carbadox peak sampl solut determin concentr sampl solut μg ml refer calibr graph feedingstuff content carbadox mg kg sampl formula gt refer graphic gt mg kg carbadox concentr sampl extract μg ml extract volum ml mass test portion premixutr prepar content carbadox mg kg sampl formula gt refer graphic gt mg kg carbadox concentr sampl extract μg ml extract volum ml premixtur prepar dilut factor premixtur prepar mass test portion valid result ident ident analyt confirm chromatographi diod arrai detector spectra sampl extract calibr solut μg ml compar chromatographi sampl extract fortifi addit amount calibr solut amount ad carbadox similar estim amount carbadox found sampl extract height carbadox peak enhanc take account amount ad dilut extract peak width half maximum height approxim origin width diod arrai detect result evalu criteria wavelength maximum absorpt sampl standard spectra record peak apex chromatogram margin determin resolv power detect system diod arrai detect typic nm nm sampl standard spectra record peak apex chromatogram part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb standard analyt nm spectra upslop apex downslop peak produc sampl extract part spectrum rang rel absorb criterion met maxima present observ point deviat spectra exce absorb spectrum apex criteria met presenc analyt confirm repeat content mg kg higher differ result parallel determin carri sampl exce rel higher result recoveri fortifi blank sampl recoveri result collabor studi collabor studi arrang feedingstuff premixtur prepar analys laboratori duplic analys perform sampl detail inform collabor studi found journal aoac volum result exclud outlier shown number laboratori number singl valu sr standard deviat repeat cvr coeffici variat repeat sr standard deviat reproduct cvr coeffici variat reproduc gt tabl gt gt tabl gt