commiss direct june establish commun method analysi offici control feedingstuff eec commiss european commun regard treati establish european econom commun regard council direct eec juli introduct commun method sampl analysi offici control feedingstuff amend act access articl thereof direct requir offici control feedingstuff carri commun method sampl analysi purpos check complianc requir aris provis laid law regul administr action qualiti composit feedingstuff commiss direct eec june eec novemb eec april eec decemb eec march eec decemb eec march establish number commun method analysi progress work make advis adopt eighth set method measur provid direct accord opinion stand committe feedingstuff adopt direct articl member state requir analys offici control feedingstuff content zinc bacitracin flavophospholipol iron copper manganes zinc carri accord method annex direct gener provis set part quot introduct quot annex direct eec except part deal prepar sampl analyz appli method annex direct articl member state januari bring forc law regul administr provis compli direct forthwith notifi commiss thereof articl direct address member state brussel june commiss finn gundelach vice presid oj oj oj oj oj oj oj oj annex determin zinc bacitracin diffus agar medium purpos scope method determin zinc bacitracin feedingstuff concentr premix lower limit determin mg kg ppm method valid presenc interf substanc larg amount copper ascorb acid principl sampl extract ph mixtur methanol water hydrochlor acid extract brought ph concentr dilut antibiot activ determin measur diffus zinc bacitracin agar medium inocul micrococcu luteu syn flavu diffus shown format zone inhibit micro organ diamet zone direct proport logarithm antibiot concentr rang antibiot concentr emploi micro organ micrococcu luteu atcc mainten stock cultur inocul micrococcu luteu agar slope cultur medium incub hour ºc store refriger ºc reinocul week agar slope prepar bacteri suspens gt pic file quot quot gt cultur media reagent cultur medium gt pic file quot quot gt assai medium gt pic file quot quot gt sodium chlorid solut dissolv sodium chlorid water dilut ml steril mixtur methanol pure water hydrochlor acid mg feedingstuff grade zinc bacitracin equival intern unit method provid establish give similar bacteri suspens commerci cultur medium similar composit give result phosphat buffer ph potassium hydrogen phosphat hpo potassium dihydrogen phosphat kh po water ml hydrochlor acid hydrochlor acid sodium hydroxid solut bromocresol purpl solut dissolv bromocresol purpl ml sodium hydroxid solut make volum ml water mix standard substanc zinc bacitracin activ standard solut weigh quantiti standard zinc bacitracin activ add ml hydrochlor acid leav stand minut add ml water adjust ph phosphat buffer ph ml make volum ml water mix ml solut prepar success dilut phosphat buffer ph solut gt pic file quot quot gt prepar extract extract concentr premix miner feed weigh quantiti sampl add ml mixtur shake briefli check ph shake minut add ml phosphat buffer ph shake minut dilut phosphat buffer obtain expect zinc bacitracin content ml protein concentr weigh quantiti sampl add ml mixtur shake briefli check ph shake minut add ml phosphat buffer ph shake minut dilut phosphat buffer obtain expect zinc bacitracin content ml feed weigh quantiti sampl expect zinc bacitracin content mg kg add ml mixtur homogen minut add ml phosphat buffer ph shake minut centrifug ml supernat solut adjust ph mean sodium hydroxid solut bromocresol purpl solut indic evapor ml rotari evapor temperatur exceed ºc dilut residu water obtain expect zinc bacitracin content ml assai solut solut prepar solut expect content ml expect content ml expect content ml mean success dilut phosphat buffer assai procedur inocul assai medium inocul assai medium bacteri suspens ºc preliminari trial plate assai medium determin quantiti bacteri suspens requir give largest clearest zone inhibit concentr zinc bacitracin prepar plate diffus agar carri plate concentr standard solut concentr assai solut concentr extract standard necessarili plate effect select plate big hole diamet mm mm centr made agar medium test carri plate consist sheet glass face aluminium plastic ring top mm diamet mm high pour plate quantiti medium inocul point give layer mm thick ml plate mm diamet set level posit bore hole place measur volum assai standard solut ml hole diamet appli concentr time determin subject evalu zone inhibit incub incub plate hour ºc evalu measur diamet zone inhibit nearest mm record measur concentr semi logarithm graph paper show logarithm concentr relat diamet zone inhibit plot fit line standard solut extract determin quot fit quot point standard lowest level sl formula gt pic file quot quot gt determin quot fit quot point standard highest level sh formula gt pic file quot quot gt similarli calcul quot fit quot point extract lowest level ul extract highest level uh substitut formula record calcul sl sh valu graph paper join give quot fit quot line standard solut similarli record ul uh join give quot fit quot line extract absenc interfer line parallel practic purpos line consid parallel valu sh sl uh ul differ line found parallel discard sl sh ul uh calcul altern formula give altern quot fit quot line gt pic file quot quot gt similarli ul uh altern fit line check parallel fact result calcul level note final report line consid parallel calcul logarithm rel activ log mean formula level gt pic file quot quot gt level gt pic file quot quot gt real activ suppos activ rel activ line consid parallel repeat determin parallel achiev calcul logarithm rel activ log mean formula result obtain consid approxim note final report repeat differ result determin carri sampl analyst exce relat highest content zinc bacitracin mg kg mg kg absolut content greater mg kg relat highest content mg kg determin flavophospholipol diffus agar medium purpos scope method determin flavophospholipol feedingstuff concentr premix lower limit determin mg kg ppm principl sampl extract dilut methanol heat reflux centrifug extract purifi treatment ion exchang resin dilut antibiot activ determin measur diffus flavophospholipol agar medium inocul staphylococcu aureu diffus shown format zone inhibit micro organ diamet zone direct proport logarithm antibiot concentr rang antibiot concentr emploi micro organ staphylococcu aureu atcc mainten stock cultur inocul staphylococcu aureu agar slope cultur medium incub hour ºc store refriger ºc arid reinocul month agar slope prepar bacteri suspens set tube stock cultur reinocul weekli incub hour ºc store refriger ºc hour assai inocul growth tube slope cultur medium incub hour ºc make suspens growth sodium chlorid solut light transmiss suspens measur nm cm cell sodium chlorid solut method provid establish give similar bacteri suspens gt pic file quot quot gt gt pic file quot quot gt prepar extract extract concentr premix miner feed weigh quantiti sampl add mg ascorb acid homogen ml methanol conic flask adjust ph mg tri hydroxymethyl aminomethan check ph indic paper stand minut readjust ph tri hydroxymethyl aminomethan boil minut reflux constant stir cool centrifug mixtur decant extract feed weigh quantiti saml flavophospholipol homegen ml methanol conic flask adjust ph mg tri hydroxymethyl aminomethan check ph indic paper stand minut readjust ph tri hydroxymethyl aminomethan boil minut reflux constant stir cool centrifug mixtur decant extract purif step omit concentr premix miner feed mix ml extract cation exchang boil minut reflux constant stir separ cation exchang centrifug filtrat mix ml extract ml methanol store solut hour ºc filter floccul mass cold insert glass wool plug bottom end glass tube pour tube ml anion exchang wash column ml methanol funnel transfer column volum filtrat ml expect flavophospholipol ml sampl feedingstuff ppm applic column dilut filtrat methanol obtain expect flavophospholipol content ml adjust flow rate ml minut discard effluent wash column ml methanol discard effluent elut flavophospholipol methanol solut potassium chlorid keep flow rate ml minut collect ml eluat graduat flask add ml water mix solut flavophospholipol content ml assai solut purif step omit dilut extract obtain point methanol obtain expect flavophospholipol content ml solut prepar solut expect content ml expect content ml expect content ml mean success dilut methanol assai procedur inocul assai medium inocul assai medium bacteri suspens ºc preliminari trial plate assai medium determin quantiti bacteri suspens requir give largest clearest zone inhibit concentr flavophospholipol ml litr prepar plate diffus agar carri plate concentr standard solut concentr assai solut concentr extract standard necessarili plate effect select plate big hole diamet mm mm centr made agar medium test carri plate consist sheet glass face aluminium plastic ring top mm diamet mm high pour plate quantiti medium give layer mm thick ml plate mm diamet set level posit layer quantiti medium inocul point give layer mm thick ml plate mm diamet set level posit bore hole place measur volum assai standard solut ml hole diamet appli concentr time determin subject evalu zone inhibit incub icub plate hour ºc evalu measur diamet zone inhibit nearest mm record measur concentr semi logarithm graph paper show logarithm concentr relat diamet zone inhibit plot fit line standard solut extract determin quot fit quot point standard lowest level sl formula gt pic file quot quot gt similarli calcul quot fit quot point extract lowest level ul extract highest level uh substitut formula record calcul sl sh valu graph paper join give quot fit quot line standard solut similarli record ul uh join give quot fit quot line extract absenc interfer line parallel practic purpos line consid parallel valu sh sl uh ul differ line found parallel discard sl sh ul uh calcul altern formula give altern quot fit quot line gt pic file quot quot gt similarli ul uh altern fit line check parallel fact result calcul level note final report line consid parallel calcul logarithm rel activ log mean formula level gt pic file quot quot gt level gt pic file quot quot gt real activ suppos activ rel activ line consid parallel repeat determin parallel achiev calcul logarithm rel activ log mean formula result obtain consid approxim note final report repeat differ result determin carri sampl analyst exce mg kg absolut content flavophospholipol mg kg relat highest content greater mg kg relat highest content greater mg kg mg kg absolut content greater mg kg relat highest content mg kg determin trace element iron cooper manganes zinc purpos scope method make determin trace element iron copper manganes zinc feedingstuff lower limit determin iron fe mg kg copper cu mg kg manganes mn mg kg zinc zn mg kg principl sampl brought solut hydrochlor acid destruct organ matter element iron copper manganes zinc determin dilut atom absorpt spectrometri reagent introductori comment prepar reagent analyt solut water free cation determin obtain doubl distil water borosil glass quartz doubl treatment ion exchang resin reagent analyt grade freedom element determin check blank experi reagent purifi place standard solut decrib commerci standard solut provid guarante check hydrochlor acid hydrochlor acid hydrochlor acid hydrofluor acid iron content mg fe litr residu evapor mg sulphat litr sulphur acid hydrogen peroxid approxim volum oxygen weight standard iron solut fe ml prepar dissolv iron wire ml hydrochlor acid add ml hydrogen peroxid make litr water work standard iron solut fe ml prepar dilut standard solut water standard copper solut cu ml prepar dissolv copper powder form ml hydrochlor acid add ml hydrogen peroxid make litr water work standard copper solut cu ml prepar dilut standard solut water dilut result solut water standard manganes solut mn ml prepar dissolv manganes powder form ml hydrochlor acid make litr water work standard manganes solut mn ml prepar dilut standard solut water dilut result solut water standard zinc solut zn ml prepar dissolv zinc strip leaf form ml hydrochlor acid make litr water work standard zinc solut zn ml prepar dilut standard solut water dilut result solut water lanthanum chorid solut prepar dissolv lanthanum oxid ml water add ml hydrochlor acid make litr water apparatu muffl furnac temperatur regul record glasswar resist borosil type recommend apparatu reserv exclus trace element determin platinum crucibl option quartz crucibl atom absorpt spectrophotomet meet requir method regard sensit precis requir rang procedur sampl organ matter ash prepar solut analysi place sampl weigh nearest mg quartz platinium crucibl note dry oven ºc introduc crucibl cold muffl furnac close furnac note gradual rais temperatur ºc minut maintain temperatur hour overnight remov carbonac materi open furnac cool note wash crucibl total ml hydrochlor acid add slowli carefulli beaker vigor reaction due green fodder fresh dri liabl larg amount veget silica retain trace element remov sampl feedingstuff modifi procedur carri oper filtrat wash filter paper insolubl residu boil water place platinum crucibl ignit muffl furnac temperatur ºc carbonac materi complet disappear cool add drop water ml hydrofluor add evapor dryness ºc silica remain residu redissolv millilitr hydrofluor add evapor dryness add drop sulphur acid heat white fume addit ml hydrochlor acid ml water heat filter solut ml volumetr flask make mark water hcl concentr proce determin point format add hydrochlor acid dropwis agit effervesc stop evapor dryness occasion stir glass rod add ml hydrochlor acid residu ml water stir glass rod left beaker cover beaker watchglass bring gentli boil maintain boil point ash dissolv filter ash free filter paper collect filtrat ml volumetr flask wash beaker filter ml hot hydrochlor acid boil water fill volumetr flask mark water hcl concentr ii residu filter appear black carbon put back furnac ash ºc ash requir hour hour complet ash appear white white dissolv residu ml hydrochlor acid evapor dryness add ml hydrochlor acid heat filter solut volumetr flask make mark water hcl concentr note determin trace element import alert risk contamin zinc copper iron reason equip prepar sampl free metal reduc gener risk contamin work dust free atmospher scrupul clean equip carefulli wash glasswar determin zinc sensit type contamin glasswar reagent dust weight sampl ash calcul approxim trace element content feedingstuff relat sensit spectrophotomet feedingstuff low trace element start sampl make final solut ml ash carri close furnac inject air oxygen temperatur pyromet exce ºc spectrophotometr determin prepar calibr solut element determin prepar work standard solut point rang calibr solut calibr solut hcl concentr case iron manganes zinc lanthanum chlorid concentr equival la trace element concentr select lie rang sensit spectrophotomet tabl show composit typic rang calibr solut depend type sensit spectrophotomet select concentr gt pic file quot quot gt gt pic file quot quot gt prepar solut analysi determin copper solut prepar point directli bring concentr rang calibr solut aliquot portion pipet ml volumetr flask made mark hydrochlor acid determin iron manganes zinc pipett aliquot portion solut prepar point ml volumetr flask add ml lanthanum chlorid solut make mark hydrochlor acid point quot observ quot blank experi blank experi includ prescrib step procedur sampl materi omit calibr solut quot quot blank measur atom absorpt measur atom absorpt calibr solut solut analyz oxid air acetylen flame wavelength gt pic file quot quot gt carri measur time miner feedingstuff sampl organ matter prior ash unnecessari proce point start paragraph evapor hydrofluor acid omit calcul result calibr curv calcul trace element concentr solut analyz express result milligram trace element kilogram sampl ppm repeat differ result zwo parallel determin carri sampl analyst exce mg kg absolut content trace element concern mg kg higher result content trace element concern mg kg mg kg absolut content trace element concern mg kg higher result content trace element concern mg kg observ gt pic file quot quot gt