fourth commiss direct decemb establish commun method analysi offici control feedingstuff eec commiss european commun regard treati establish european econom commun regard council direct juli introduct commun method sampl analysi offici control feedingstuff amend direct eec juli 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 direct eec june eec novemb eec april establish number commun method analysi progress work make advis adopt fourth set method measur provid direct accord opinion manag committe feedingstuff adopt direct articl member state requir analys offici control feedingstuff moistur content anim veget fat oil magnesium crude fibr content feedingstuff carri method annex direct gener provis set part introduct annex commiss direct eec june applic method annex direct articl member state requir analys offici control feedingstuff content retinol vitamin thiamin aneurin vitamin ascorb dehydroascorb acid vitamin carri method annex ii direct gener provis set part introduct annex commiss direct eec june except part deal prepar sampl analys applic method annex ii direct articl member state januari bring forc law regul administr provis compli direct forthwith notifi commiss thereof articl direct address member state brussel decemb commiss presid mansholt oj oj oj oj oj annex determin moistur anim veget fat oil purpos scope method make determin water volatil substanc content anim veget fat oil principl sampl dri constant weight ºc loss mass determin weigh apparatu flat bottom dish corros resist materi cm diamet approxim cm high mercuri thermomet strengthen bulb expans tube top end graduat approxim ºc ºc approxim cm length sand bath electr hot plate desicc effici dry agent analyt balanc procedur weigh nearest mg approxim homogen sampl dry weigh dish thermomet heat sand bath hot plate stir continu thermomet temperatur reach ºc minut reduc heat watch frequenc bubbl rise bottom dish temperatur exce ºc continu stir scrape bottom dish bubbl stop form order ensur complet elimin moistur reheat time ºc ºc cool ºc success heat leav cool room temperatur desicc weigh repeat oper loss mass success weigh longer exce mg increas mass sampl repeat heat oxid fat case calcul result weigh carri immedi mass began increas calcul result moistur content percentag sampl formula gt pic file quot quot gt mass gramm test sampl mass gramm dish content heat mass gramm dish content heat result lower record quot lower quot repeat differ moistur result parallel determin carri sampl exce absolut determin magnesium atom absorpt spectrophotometri purpos scope method make determin quantiti magnesium feed stuff determin magnesium content lower principl sampl ash dissolv dilut hydrochlor acid organ substanc dissolv directli dilut hydrochlor acid solut dilut magnesium content determin atom absorpt spectrophotometri nm comparison standard solut reagent hydrochlor acid concentr hydrochlor acid magnesium ribbon wire magnesium sulphat heptahydr dri room temperatur strontium salt solut chlorid nitrat strontium src sr standard magnesium solut weigh nearest mg magnesium previous oxid coat carefulli remov quantiti magnesium sulphat heptahydr place ml graduat flask add ml hydrochlor acid leav dissolv make ml water ml solut mg magnesium apparatu platinum silica porcelain ash crucibl thermostat control electr muffl furnac atom absorpt spectrophotomet procedur prepar sampl solut feed stuff compos exclus miner substanc weigh nearest mg sampl ml graduat flask ml water add ml hydrochlor acid bring boil liquid gentli boil minut leav cool make volum water mix filter dry beaker dry pleat filter discard ml filtrat presenc silica treat sampl suffici quantiti ml hydrochlor acid evapor dryness water bath transfer oven ºc hour proce sentenc feed stuff compos predominantli miner substanc weigh nearest mg sampl crucibl ash ºc muffl furnac ash free carbonac particl obtain leav cool order elimin silica add ash suffici quantiti ml hydrochlor acid evapor dryness water bath transfer oven ºc hour treat residu ml hydrochlor acid transfer ml graduat flask warm water leav cool make volum water mix filter dry beaker dry pleat filter discard ml filtrat feedingstuff compos predominantli organ substanc weigh nearest mg sampl crucibl ash ºc muffl furnac ash free carbonac particl obtain treat ash ml hydrochlor acid evapor dryness water bath dry hour oven ºc order render silica insolubl treat ash ml hydrochlor acid transfer ml graduat flask warm water bring boil leav cool make volum water mix filter dry beaker dry pleat filter discard ml filtrat measur atom absorpt dilut standard solut water prepar refer solut increas concentr optim measur rang spectrophotomet add solut ml strontium salt solut make volum ml water dilut water aliquot part filtrat obtain obtain magnesium concentr limit concentr refer solut hydrochlor acid concentr solut exce add ml strontium salt solut make volum ml water measur absorpt solut determin refer solut nm calcul result calcul quantiti magnesium sampl relat refer solut express result percentag sampl repeat differ result parallel determin carri sampl exce rel determin crude fibr purpos scope method make determin feed stuff quantiti fat free organ substanc insolubl acid alkalin media convention crude fibr principl sampl defat treat success boil solut sulphur acid potassium hydroxid concentr residu separ filtrat presenc asbesto wash dri weigh ash ºc loss weight result ash correspond crude fibr present test sampl reagent sulphur acid treat asbesto add asbesto type gooch crucibl approxim time weight dilut hydrochlor acid volum hydrochlor acid volum water boil mixtur approxim minut leav cool filter buchner funnel wash residu water hydrochlor acid disappear wash water aceton dry asbesto dry oven ash hour ºc leav cool stopper flask asbesto treat time meet specif blank test antifoam emuls silicon potassium hydroxid solut hydrochlor acid aceton diethyl ether apparatu beaker ml capac measur mark ml level porcelain disc approxim mm diamet approxim mm thick perfor approxim hole approxim mm diamet rubber stopper vacuum flask approxim litr capac measur mark ml level fit glass funnel mm diamet filter plate approxim mm diamet approxim mm thick slant edg fit cone funnel perfor approxim hole approxim mm diamet cover wire mesh mesh size approxim mm plate wire mesh resist acid alkali platinum silica ash crucibl thermostat control electr muffl furnac desicc asbesto filter suspend asbesto ml water filter vacuum filter plate cover wire mesh funnel vacuum flask collect filtrat filter filter discard filtrat procedur weigh nearest mg sampl treat asbesto beaker add ml sulphur acid drop antifoam emuls bring rapidli boil leav boil minut constant volum cover beaker cool devic ml round bottom flask cold water circul stop boil ad approxim ml cold water filter immedi vacuum asbesto filter previous prepar shown wash residu lot approxim ml hot water obtain final volum filtrat ml transfer residu quantit beaker fit porcelain disc regul boil add ml potassium hydroxid solut bring rapidli boil leav boil minut add approxim ml cold water filter immedi vacuum fresh asbesto filter previous prepar shown wash residu hot water wash water neutral test litmu paper time aceton approxim ml aceton transfer residu quantit ash crucibl break dry constant weight dry oven ºc leav cool desicc weigh rapidli place crucibl muffl furnac leav ash minut ºc leav cool desicc weigh rapidli carri blank test appli procedur treat asbesto sampl loss weight result ash asbesto exce mg calcul result crude fibr content percentag sampl ratio gt pic file quot quot gt loss weight ash determin loss weight ash blank test repeat differ result parallel determin carri sampl exce absolut crude fibr content lower rel crude fibr content equal greater observ feedingstuff oil fat defat prior analysi diethyl ether place test sampl weigh nearest mg asbesto filter cover time approxim ml diethyl ether time filter carefulli vacuum transfer defat test sampl asbesto quantit beaker continu analysi shown feedingstuff oil fat extract directli defat shown defat time acid attack wash residu wash attack residu time aceton ml time ml diethyl ether transfer residu quantit beaker continu analysi shown paragraph treatment potassium hydroxid solut feedingstuff rich calcium calcium place test sampl weigh nearest mg beaker ml hydrochlor acid leav stand cool temperatur minut filter immedi wash cold water filtrat aid asbesto boil sulphur acid filtrat prove difficult dilut suspens aceton proce shown annex ii determin retinol vitamin purpos scope method make determin quantiti retinol vitamin feedingstuff concentr premix lower limit determin iu kg highli pigment feed iu kg product classifi group presum retinol content group content lower iu kg group content equal greater iu kg principl sampl hydrolis hot potassium hydroxid solut ethanol medium presenc antioxid nitrogen atmospher mixtur extract dichlorethan extract evapor dryness treat light petroleum solut chromatograph column aluminium oxid iu retinol group product chromatographi requir case group product retinol determin spectrophotometri nm develop colour complex carr price reaction group product spectrophotometri uv nm reagent analys product group ethanol sodium ascorb solut purifi nitrogen potassium hydroxid solut potassium hydroxid solut potassium hydroxid solut dichlorethan light petroleum boil rang ºc purifi stir ml light petroleum ml lot concentr sulphur acid acid remain colourless remov acid wash light petroleum success ml water ml sodium hydroxid solut time ml water remov aqueou layer dry light petroleum hour activ carbon anhydr sodium sulphat filter distil aluminium oxid standard brockmann ash hour ºc cool desicc brown glass bottl fit ground glass stopper chromatographi moisten place brown glass bottl aluminium oxid ml water seal stopper reheat minut boil water bath shake leav cool verifi activ aluminium prepar subject quantiti retinol ca iu procedur check recoveri basic aluminium oxid degre activ woelm merck equival pure diethyl ether remov peroxid trace water chromatographi column basic aluminium oxid aluminium oxid ml diethyl ether light petroleum solut diethyl ether sodium sulphid solut molar glycerin prepar sodium sulphid exclus analys group product crystalliz benzen chloroform remov ethanol phosgen trace water chromatographi column basic aluminium oxid aluminium oxid ml chloroform advis chromatograph ml eluat time carr price reagent stir approxim antimoni trichlorid desicc ml chloroform solut satur slight deposit antimoni trichlorid problem add ml acet anhydrid refriger brown glass bottl ground glass stopper solut week retinol standard spectrophotometr exclus analys group product isopropanol chromatographi apparatu water bath vacuum evapor apparatu round flask capac glass chromatographi tube length mm intern diamet mm spectrophotomet mm cell measur uv requir silica cell uv lamp suitabl nm procedur oper carri direct light brown glass equip test sampl fine divid sampl test sampl proport presum retinol content concentr content greater iu premix content iu miner mixtur group product immedi place test sampl ml flask ground glass stopper hydrolysi extract add success test sampl ml ethanol ml sodium ascorb solut ml potassium hydroxid solut ml sodium sulphid solut heat minut ºc reflux condens leav cool stream water add ml ethanol ml dichlorethan pipett shake vigor decant supernat liquid decant contain add contain ml potassium hydroxid solut shake second leav stand layer separ collect dichlorethan layer lower layer decant contain add ml potassium hydroxid solut shake second leav stand layer separ collect dichlorethan layer decant contain wash time ml lot water free alkali phenolphthalein test collect dichlorethan layer remov trace water strip filter paper evapor dryness aliquot part solut vacuum water bath ºc rapidli treat residu ml light petroleum group product chromatograph shown group product transfer solut ml graduat flask make volum light petroleum mix measur optic densiti shown chromatographi group product fill chromatographi tube height mm aluminium oxid previous slurri light petroleum place tube solut obtain immedi add ml light petroleum elut success ml lot light petroleum solut diethyl ether pressur partial vacuum rate flow drop milk feed product tendenc agglomer swell doubl quantiti reagent shown paragraph sodium ascorb ad hydrolysi carri nitrogen atmospher caroten elut retinol gener elut light petroleum solut diethyl ether elut uv light irradi column mercuri lamp fluoresc zone retinol separ yellow xanthophyl zone collect eluat fraction retinol erlenmey flask group product chromatographi carri optic densiti measur obtain conform requir chromatographi prove place chromatographi column aliquot part solut light petroleum obtain approxim iu retinol chromatograph shown measur optic densiti group product evapor dryness vacuum eluat retinol obtain treat residu ml benzen ml solut add ml carr price reagent blue colour develop measur optic densiti spectrophotomet nm second reaction begun determin retinol content refer standard curv obtain benzen solut increas retinol standard concentr treat carr price reagent iu retinol standard ml benzen ml carr price reagent standard curv check regularli frequent standard freshli prepar carr price reagent solut group product aliquot part solut light petroleum obtain approxim iu retinol evapor dryness vacuum treat residu ml isopropanol measur optic densiti spectrophotomet nm absorpt maximum locat nm retinol content solut calcul iu retinol ml ratio optic densiti ratio differ appreci measur optic densiti preced chromatographi accord method measur optic densiti carri chromatographi show abovement ratio differ appreci determin carri accord method group product calcul result calcul retinol content sampl take account weight test sampl dilut carri analysi express result iu retinol kg feedingstuff kg concentr premix gt pic file quot quot gt repeat differ result parallel determin carri sampl exce rel retinol content lower iu kg iu content iu kg rel content iu kg iu content iu kg rel content greater iu kg determin thiamin vitamin aneurin purpos scope method make determin quantiti thiamin aneurin vitamin feed stuff concentr premix lower limit determin ppm principl solut treat hot dilut sulphur acid hydrolys enzymat solut obtain subject alkalin oxid thiochrom form extract isobutanol determin fluorimetri reagent ml standard thiamin solut dissolv mg thiamin hydrochlorid previoulsli dri vacuum constant weight ml sulphur acid store cool dark place solut month sulphur acid pure sodium bisulphit potassium ferricyanid solut potassium hydroxid solut oxid mixtur mix ml potassium ferricyanid solut ml potassium hydroxid solut mixtur hour isobutanol sodium acet solut multienzymat prepar proteas phosphatas amylas claras ethanol apparatu water bath centrifug rpm ml tube fit ground glass stopper fluorimet procedur enzymat hydrolysi place ml graduat flask ident amount fine divid sampl approxim thiamin ml sulphur acid add flask ml standard solut intern standard shake flask vigor place boil water bath minut shake occasion leav cool approxim ºc add flask ml sodium acet solut multienzymat prepar leav stand minut room temperatur add ml sodium acet solut make volum water homogen filter collect filtrat discard ml prepar solut refer solut place centrifug tube ml filtrat approxim mg sodium bisulphit immers tube boil water bath minut leav cool room temperatur solut intern standard sampl place ml filtrat centrifug tube ml filtrat centrifug tube oxid add solut ml oxidis mixtur minut ml isobutanol stopper tube shake vigor second leav stand minut centrifug separ layer tube transfer ml supernat isobutanol layer ml graduat flask make volum ethanol homogen extract measur fluoresc carri measur wavelength fluorimet optim respons fluoresc thiochrom irradi approxim nm adjust instrument extract measur intens fluoresc extract calcul result thiamin content mg kg sampl ratio gt pic file quot quot gt intens fluoresc extract intern standard intens fluoresc extract sampl weight test sampl amount thiamin ad test sampl intern standard repeat differ result parallel determin carri sampl exce rel content lower mg kg rel content equal greater mg kg determin ascorb acid dehydroascorb acid vitamin purpos scope method make determin total quantiti ascorb dehydroascorb acid vitamin feedingstuff concentr premix lower limit determin ppm product classifi group presum vitamin content group content lower kg group content equal greater kg principl sampl suspend dilut solut metaphosphor acid extract chloroform aqueou phase treat solut dichlorophenol indophenol order transform ascorb acid dehydroascorb acid solut dinitrophenylhydrazin hydrazon form extract mixtur ethyl acet glacial acet acid aceton solut chromatograph column silica gel eluat evapor dryness residu dissolv dilut sulphur acid optic densiti solut measur spectrophotomet nm group product eluat result chromatographi column subject thin layer chromatographi isol hydrazon reagent standard solut ascorb acid dissolv mg ascorb acid approxim ml metaphosphor acid solut make ml water prepar immedi metaphosphor acid solut grind mortar dissolv water metaphosphor acid make ml water ºc stabl week chloroform solut dichlorophenol indophenol prepar immedi filtrat aid equival acid solut dinitrophenylhydrazin dissolv dinitrophenylhydrazin ml dilut sulphur acid ml sulphur acid dilut make ml water store cool temperatur solut week nitrogen carbon dioxid mixtur ethyl acet glacial acet acid aceton volum mixtur dichloromethan glacial acet acid volum silica gel particl size mm stahl grade silica gel thin layer chromatographi dilut sulphur acid place ml water ml graduat flask make volum sulphur acid elut solvent thin layer chromatographi mix ml diethyl ether ml ethyl acet ml acet acid renew chromatographi apparatu water bath fit thermostat set ºc centrifug rpm ml tube fit ground glass stopper rotari vacuum evapor ml flask glass chromatographi tube length mm intern diamet mm sinter disc allihn tube spectrophotomet colorimet filter mm cell apparatu thin layer chromatographi silica gel plate coat depth mm readi made plate dry plate hour dry oven ºc leav cool desicc hour dry oven set ºc procedur extract place ml graduat flask ground glass stopper ident quantiti fine divid sampl vitamin add flask ml standard solut mix shake gentli intern standard add flask ml chloroform ml metaphosphor acid solut ºc shake briefli leav stand minut add ml water stopper flask shake vigor second leav stand minut water bath centrifug separ aqueou phase chloroform phase carri oper simultan aqueou extract intern standard oxid pipett transfer ml supernat aqueou solut slightli cloudi obtain reaction tube fit ground glass stopper add ml dichlorophenol indophenol solut mix red colour develop remain minut add approxim mg filtrat aid shake filter dry pleat filter filtrat necessarili clear reaction dinitrophenylhydrazin hydrazon extract pipett transfer ml filtrat obtain centrifug tube add ml dinitrophenylhydrazin solut mix pass stream nitrogen carbon dioxid rapidli tube stopper tube immers approxim hour overnight water bath add ml water ml ethyl acet glacial acet acid aceton mixtur approxim mg filtrat aid stopper tube shake vigor second centrifug place ml supernat phase evapor flask evapor reduc pressur rotari evapor oili residu obtain dissolv residu ml ethyl acet glacial acet acid aceton mixtur heat ºc leav cool add ml dichloromethan glacial acet acid mixtur mix chromatographi column fill chromatographi tube level mm dichloromethan glacial acet acid mixtur suspend shake vigor silica gel ml dichloromethan glacial acet acid mixtur pour suspens tube leav stand compress nitrogen low pressur decant tube solut obtain rins flask small quantiti dichloromethan glacial acet acid mixtur decant tube fill mixtur proce wash column mixtur lot approxim ml colourless eluat obtain discard part eluat colour yellow elut reddish zone top column ethyl acet glacial acet acid aceton mixtur collect eluat evapor dryness group product content vitamin lower kg dissolv residu ml ethyl acet glacial acet acid aceton mixtur chromatograph immedi thin layer shown group product content vitamin equal greater kg treat oili residu ml dilut sulphur acid shake vigor dissolv residu complet measur optic densiti shown thin layer chromatographi carri duplic oper place thin line plate ml solut obtain elut solvent develop minut tank satur solvent vapour pinkcolour hydrazon zone separ leav dry open mark limit pink zone scrape zone spatula quantit transfer powder chromatographi tube elut success ml ml ethyl acet glacial acet acid aceton mixtur collect eluat small flask part colourless evapor dryness treat oili residu ml dilut sulphur acid shake vigor dissolv residu complet measur optic densiti measur optic densiti measur optic densiti spectrophotomet nm minut dissolv residu sulphur acid carri measur comparison dilut sulphur acid blank test carri blank test appli procedur sampl calcul result vitamin content sampl kg ratio gt pic file quot quot gt optic densiti blank optic densiti intern standard solut optic densiti sampl solut weight gramm test sampl repeat differ result parallel determin carri sampl exce rel vitamin content lower kg rel content equal greater kg