commiss direct decemb amend direct eec eec eec 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 greec articl thereof commiss direct eec eec eec set method analysi crude oil fat virginiamycin zinc bacitracin replac method method reflect advanc scientif technic knowledg measur provid direct accord opinion stand committe feedingstuff adopt direct articl annex direct eec part iv determin crude oil fat replac annex direct articl annex ii direct eec part determin virginiamycin diffus agar medium replac annex ii direct articl annex direct eec part determin zinc bacitracin diffus agar medium replac annex iii direct articl member state june latest bring forc law regul administr provis compli direct forthwith inform commiss thereof articl direct address member state brussel decemb commiss poul dalsag member commiss oj oj oj oj annex determin crude oil fat purpos scope method make determin content crude oil fat feedingstuff cover analysi oil seed oleagin fruit defin council regul eec septemb depend natur feedingstuff method method applic sraight feedingstuff plant origin except oil fat total extract light petroleum prior hydrolysi gluten yeast soya potato protein method applic compound feedingstuff except milk powder oil fat total extract light petroleum prior hydrolysi method applic straight feedingstuff anim origin feedingstuff mention point except method principl method sampl extract light petroleum solvent distil residu dri weigh method sampl treat heat hydrochlor acid mixtur cool filter residu wash dri submit determin method reagent light petroleum boil rang bromin residu evapor mg ml sodium sulphat anhydr hydrochlor acid filtrat aid kieselgur hyflo supercel apparatu extract apparatu fit siphon soxhlet apparatu reflux rate produc cycl hour siphon type reflux rate ml minut extract thimbl free matter solubl light petroleum poros consist requir point dry oven vacuum oven set air oven set procedur method point weigh sampl nearest mg transfer extract thimbl cover fat free wad cotton wool place thimbl extractor extract hour light petroleum collect light petroleum extract dry weigh flask fragment pumic stone distil solvent dry evapor residu maintain flask half hour dry oven leav cool dessicc weigh dry minut ensur weight oil fat remain constant loss weight success weigh mg method weigh sampl nearest mg point place ml beaker ml conic flask add ml hydrochlor acid fragment pumic stone cover beaker watch glass fit conic flask reflux condens bring mixtur gentl boil low flame hot plate hour product stick side contain cool add quantiti filtrat aid suffici prevent loss oil fat filtrat filter moisten fat free doubl filter paper wash residu cold water neutral filtrat obtain check filtrat oil fat presenc sampl extract light petroleum method hydrolysi place doubl filter paper residu watch glass dry half hour oven place doubl filter paper dry residu extract thimbl cover fat free wad cotton wool place thimbl extractor proce paragraph point express result express weight residu percentag sampl repeat differ result parallel determin carri sampl analyst exce absolut content crude oil fat lower relat highest result content absolut content observ product high content oil fat difficult crush unsuit draw homogen reduc test sampl proce weigh sampl nearest mg mix anhydr sodium sulphat extract light petroleum point make extract obtain ml light petroleum homogen ml solut place small dry weigh flask fragment pumic stone distil solvent dry proce paragraph point elimin solvent extract residu left thimbl crush residu fine mm return extract thimbl add sodium sulphat proce paragraph point calcul content oil fat percentag sampl formula mass gram residu extract aliquot part extract mass gram residu extract product low oil fat test sampl increas oil fat undergo subsequ qualiti test replac fragment pumic stone glass bead oil fat undergo subsequ qualiti test replac fragment pumic stone glass bead annex ii determin virginiamycin diffus agar medium purpos scope method determin virginiamycin feedingstuff premix lower limit determin mg kg ppm principl sampl extract methanol solut tween extract decant centrifug dilut antibiot activ determin measur diffus virginiamycin agar medium inocul micrococcu luteu diffus shown format zone inhibit micro organ diamet zone direct proport logarithm antibiot concentr rang antibiot concentr emploi micro organ micrococcu luteu atcc nctc ncib mainten stock cultur inocul tube slope cultur medium micrococcu luteu incub hour store cultur refriger reinocul week prepar bacteri suspens harvest growth recent prepar agar slope ml sodium clorid solut suspens inocul ml cultur medium contain roux flask incub hour harvest growth ml sodium chlorid solut mix dilut suspens sodium chlorid solut light transmiss suspens measur nm cm cell sodium chlorid solut suspens week cultur media reagent cultur assai medium meat pepton trypton yeast extract meat extract glucos agar water ml ph steril phosphat buffer ph potassium hydrogen phosphat hpo potassium dihydrogen phosphat kh po water ml sodium chlorid solut dissolv sodium chlorid water dilut ml steril methanol mixtur phosphat buffer methanol tween methanol solut dissolv tween methanol dilut methanol ml standard substanc virginiamycin activ standard solut dissolv accur weigh quantiti standard substanc methanol dilut methanol give stock solut mg virginiamycin ml store stopper flask solut stabl dai stock solut prepar success dilut mixtur solut mg ml mg ml mg ml mg ml prepar extract assai solut extract product virginiamycin content mg kg weigh quantiti sampl add ml solut shake minut leav settl centrifug ml supernat solut evapor ml rotari evapor temperatur exceed dilut residu mixtur obtain expect virginiamycin content mg ml product virginiamycin content greater mg kg weigh quantiti sampl exceed mg virginiamycin add ml solut shake minut leav settl centrifug dilut supernat solut mixtur obtain expect virginiamycin content ml assai solut solut prepar solut expect content ml expect content mg ml expect content mg ml mean success dilut mixtur assai procedur inocul assai medium inocul assai medium bacteri suspens preliminari trial plate medium determin quantiti bacteri suspens requir give largest clearest zone inhibit concentr virginiamycin 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 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 sl determin fit quot point standard highest level sh formula sh 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 sl sh similarli ul uh criteria parallel satisfi fact result calcul level note final report line consid parallel calcul logarithm rel activ log mean formula depend level assess parallel level log level log log activ sampl extract activ relev standard rel activ found rang repeat assai make adjust extract concentr standard solut rel activ brought requir rang result obtain consid approxim note final report line consid parallel repeat determin parallel achiev determin consid unsatisfactori express result milligram virginiamycin kilogram feedingstuff repeat differ result parallel determin carri sampl analyst exce mg kg absolut content virginiamycin mg kg relat highest content mg kg mg kg absolut content mg kg relat highest content mg kg mg virginiamycin equival uk unit method provid establish give similar bacteri suspens commerci cultur medium similar composit give result annex iii determin zinc bacitracin diffus agar medium purpos scope method determin zinc bacitracin feedingstuff premix lower limit determin mg kg ppm principl sampl extract ph mixtur methanol water hydrochlor acid sodium sulphid solut addit sodium sulphid precipit solubl copper salt interfer assai extract brought ph concentr dilut antibiot activ determin measur diffus zinc bacitracin agar medium inocul micrococcu luteu flavu diffus shown format zone inhibit micro organ diamet zone direct proport logarithm antibiot concentr rang antibiot concentr emploi micro organ micrococcu luteu flavu atcc mainten stock cultur inocul tube slope cultur medium micrococcu luteu flavu incub hour store cultur refriger reinocul week prepar bacteri suspens harvest growth recent prepar agar slope ml sodium chlorid solut suspens inocul ml cultur medium contain roux flask incub hour harvest growth ml sodium chlorid solut mix dilut suspens sodium chlorid solut light transmiss suspens measur nm cm cell sodium chlorid solut suspens week cultur media reagent cultur medium meat pepton trypton yeast extract meat extract glucos agar water ml ph steril assai medium trypton yeast extract meat extract glucos agar tween ml water ml ph steril sodium chlorid solut dissolv sodium chlorid water dilut ml steril mixtur methanol water hydrochlor acid phosphat buffer ph potassium hydrogen phosphat hpo potassium dihydrogen phosphat kh po water ml hydrochlor acid hydrochlor acid sodium hydroxid solut sodium sulphid 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 ml make volum ml water mix ml solut prepar success dilut phosphat buffer solut ml ml ml ml prepar extract assai solut extract premix miner feed weigh quantiti sampl add ml mixtur ml sodium sulphid solut shake briefli check ph shake minut add ml phosphat buffer shake minut centrifug suitabl aliquot supernat solut adjust ph mean hydroxid solut ph meter bromocresol purpl solut indic dilut phosphat buffer obtain expect zinc bacitracin content ml protein concentr weigh quantiti sampl add ml mixtur ml sodium sulphid solut shake briefli check ph shake minut add ml phosphat buffer shake minut centrifug suitabl volum supernat solut adjust ph mean sodium hydroxid solut ph meter bromocresol purpl solut indic evapor approxim half volum rotari evapor temperatur exceed dilut phosphat buffer obtain expect zinc bacitracin content ml feed weigh quantiti sampl expect zinc bacitracin content mg kg add mixtur ml mixtur ml sodium sulphid solut homogen minut add ml phosphat buffer shake minut centrifug ml supernat solut adjust ph mean sodium hydroxid solut ph meter bromocresol purpl solut indic evapor ml rotari evapor temperatur exceed dilut residu phosphat buffer 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 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 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 sl determin fit quot point standard highest level sh formula sh 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 sl sh similarli ul uh criteria parallel satisfi fact result calcul level note final report line consid parallel calcul logarithm log rel activ mean formula depend level assess parallel level log level log log activ sampl extract activ relev standard rel activ found rang repeat assai make adjust extract concentr standard solut rel activ brought requir rang result obtain consid approxim note final report line consid parallel repeat determin parallel achiev satisfactori determin obtain express result milligram zinc bacitracin kilogram feedingstuff repeat differ result determin carri sampl analyst exce mg kg absolut content zinc bacitracin mg kg relat highest content mg kg mg kg absolut content mg kg relat highest content mg kg mg feedingstuff grade zinc bacitracin equival intern unit method provid establish give similar bacteri suspens commerci cultur medium similar composit give result 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 ml make volum ml water mix ml solut prepar success dilut phosphat buffer solut ml ml ml ml prepar extract assai solut extract premix miner feed weigh quantiti sampl add ml mixtur ml sodium sulphid solut shake briefli check ph shake minut add ml phosphat buffer shake minut centrifug suitabl aliquot supernat solut adjust ph mean hydroxid solut ph meter bromocresol purpl solut indic dilut phosphat buffer obtain expect zinc bacitracin content ml protein concentr weigh quantiti sampl add ml mixtur ml sodium sulphid solut shake briefli check ph shake minut add ml phosphat buffer shake minut centrifug suitabl volum supernat solut adjust ph mean sodium hydroxid solut ph meter bromocresol purpl solut indic evapor approxim half volum rotari evapor temperatur exceed dilut phosphat buffer obtain expect zinc bacitracin content ml feed weigh quantiti sampl expect zinc bacitracin content mg kg add mixtur ml mixtur ml sodium sulphid solut homogen minut add ml phosphat buffer shake minut centrifug ml supernat solut adjust ph mean sodium hydroxid solut ph meter bromocresol purpl solut indic evapor ml rotari evapor temperatur exceed dilut residu phosphat buffer 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 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 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 fit quot point standard lowest level sl formula sl determin fit quot point standard highest level sh formula sh similarli calcul fit quot point extract lowest level ul extract highest level uh substitut formula record calcul sl sh valu graph paper join give fit quot line standard solut similarli record ul uh join give 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 fit quot line sl sh similarli ul uh criteria parallel satisfi fact result calcul level note final report line consid parallel calcul logarithm log rel activ mean formula depend level assess parallel level log level log log activ sampl extract activ relev standard rel activ found rang repeat assai make adjust extract concentr standard solut rel activ brought requir rang result obtain consid approxim note final report line consid parallel repeat determin parallel achiev satisfactori determin obtain express result milligram zinc bacitracin kilogram feedingstuff repeat differ result determin carri sampl analyst exce mg kg absolut content zinc bacitracin mg kg relat highest content mg kg mg kg absolut content mg kg relat highest content mg kg mg feedingstuff grade zinc bacitracin equival intern unit method provid establish give similar bacteri suspens commerci cultur medium similar composit give result