commiss direct june establish commun method analysi offici control feed stuff eec commiss european commun regard treati establish european econom commun regard council direct juli introduct commun method sampl analysi offici control feed stuff articl thereof direct requir offici control feed stuff carri commun method sampl analysi purpos check complianc requir aris provis laid law regul administr action qualiti composit feed stuff method analysi establish establish method determin hydrocyan acid calcium carbon crude ash ash insolubl hcl chlorin chlorid mustard oil lactos potassium sodium sugar theobromin urea determin alkaloid lupin estim ureas activ product deriv soya stage process measur provid direct accord opinion stand committe feed stuff adopt direct articl member state requir analys offici control feed stuff level hydrocyan acid calcium carbon crude ash ash insolubl hcl chlorin chlorid mustard oil lactos potassium sodium sugar theobromin urea determin alkaloid lupin estim ureas activ product deriv soya carri method set annex direct oj articl member state juli bring forc law regul administr provis compli direct forthwith inform commiss thereof articl direct address member state brussel june commiss presid franco malfatti annex method analysi compon feed stuff introduct method analysi compon feed stuff gener applic straight compound feed stuff feed stuff reason inher composit characterist requir individu method analysi case provid head quot observ quot descript method method determin compon feed stuff choic method left test laboratori concern method certif analysi prepar sampl analysi essenti chemic analysi carri homogen sampl carri macroscop microscop determin determin moistur sampl state reach laboratori order satisfi requir sampl divid part part prepar chemic analysi divid sampl mean mechan apparatu hand carefulli mix clean dry surfac case advis method quarter consist take sampl turn opposit section final remov analysi portion weigh approxim crush sampl pass siev round mm mesh transfer sampl immedi dry contain air tight fit seal sampl moist pre dri bring moistur content level dry sampl suitabl temperatur adequ length time reagent apparatu special instrument apparatu requir special standard mention descript method analysi consid mention apparatu instrument form part everydai equip test laboratori water mention connect dilut wash mean distil water similarli solut reagent mention indic mean solut distil water express result result certif analysi averag obtain basi test subject special provis express percentag origin sampl reach laboratori result signific figur accuraci method analysi determin hydrocyan acid purpos scope method make determin level hydrocyan acid free combin form glycosid feed stuff product deriv flax seed manioc flour speci bean principl sampl suspend water hydrocyan acid releas action enzym entrain steam distil collect specif volum acidifi silver nitrat solut silver cyanid separ filtrat excess silver nitrat titrat solut ammonium thiocyan reagent suspens sweet almond crush twenti blanch sweet almond ml water ºc check hydrocyan acid ml suspens sodium picrot paper carri blank test paragraph solut sodium acet neutral phenolphthalein anti foam emuls silicon nitric acid silver nitrat solut ammonium thiocyan solut satur solut ammonium ferric sulphat ammonia apparatu oven thermostat set ºc apparatu distil entrain steam fit condens curv extens piec ml flat bottom flask ground glass stopper oil bath burett graduat ml procedur weigh sampl nearest mg place litr flat bottom flask add ml water ml sweet almond suspens stopper flask transfer oven sixteen hour ºc cool room temperatur add ml water ml sodium acet solut drop anti foam emuls connect flask steam distil apparatu place oilbath brought temperatur slightli ºc distil ml liquid pass power current steam flask gentli heat oil bath collect distil erlenmey flask protect light ml silver nitrat solut ml nitric acid make condens extens piec immers silver nitrat solut transfer content erlenmey flask ml volumetr flask make volum water stir filter remov ml filtrat add approxim ml solut ammonium ferric sulphat back titrat excess silver nitrat solut ammonium thiocyan burett graduat ml blank test requir carri appli procedur ml sweet almond suspens omit sampl analys calcul result blank test silver nitrat solut consum subtract volum consum distil sampl ml agno correspond mg hcn express result percentag sampl observ sampl larg quantiti sulphid bean black precipit silver sulphid form filter silver cyanid deposit format precipit loss silver nitrat solut volum subtract volum calcul hcn content proce treat deposit left filter ml ammonia order dissolv silver cyanid wash residu dilut ammonia determin silver content convert obtain ml silver nitrat solut sampl hcn content determin titrat acidifi ammoniac filtrat nitric acid determin calcium purpos scope method make determin total calcium content feed stuff principl calcium ash ash treat hydrochlor acid calcium precipit calcium oxal dissolv precipit sulphur acid titrat oxal acid form solut potassium permangant reagent hydrochlor acid nitric acid sulphur acid ammonia cold satur solut ammonium oxal solut citric acid solut ammonium chlorid solut bromo cresol green solut potassium permangan apparatu electr muffl furnac air circul thermostat platinum silica porcelain crucibl ash glass filter crucibl poros procedur weigh approxim sampl nearest mg calcin ºc transfer ash ml beaker add ml hydrochlor acid ml water drop nitric acid bring boil boil point thirti minut cool transfer solut ml volumetr flask rins bring volum mark water homogenis filter pipett remov ml beaker aliquot amount mg calcium assum calcium content add ml citric acid solut ml ammonium chlorid solut make volum approxim ml water bring boil add ten drop bromo cresol green solut ml warm solut ammonium oxal precipit form dissolv ad drop hydrochlor acid neutralis slowli ammonia stir continu ph read obtain indic colour place beaker boil water bath thirti minut precipit form settl remov beaker water bath leav stand hour filter filter crucibl wash beaker crucibl water excess ammonium oxal complet remov absenc chlorid wash water suffici wash dissolv precipit filter ml warm sulphur acid rins crucibl warm water make filtrat approxim ml bring temperatur ºc titrat drop drop solut potassium permangan pink colour obtain last minut calcul result ml potassium permangan correspond mg calcium express result obtain percentag sampl observ low content calcium proce filter calcium oxal precipit ash free filter paper wash dry filter ash ºc platinum crucibl redissolv residu drop sulphur acid evapor dry calcin ºc weigh weight calcium sulphat obtain calcium content aliquot amount sampl sampl consist sole miner substanc dissolv hydrochlor acid ash case product calcium aluminium phosphat difficult dissolv acid melt alkalin process dissolv mix sampl analys platinum crucibl mixtur time weight consist equal amount potassium carbon sodium carbon heat carefulli mixtur complet melt cool dissolv hydrochlor acid magnesium content sampl high precipit calcium oxal time determin carbon purpos scope method make determin amount carbon convention express calcium carbon feed stuff case iron carbon special method principl carbon decompos hydrochlor acid carbon dioxid releas collect graduat tube volum compar releas condit quantiti calcium carbon reagent hydrochlor acid calcium carbon sulphur acid approxim colour methyl red apparatu scheibler dietrich apparatu diagram equival apparatu procedur sampl carbon content weigh portion sampl shown product carbon express calcium carbon product carbon express calcium carbon product place portion sampl special flask apparatu fit small tube unbreak materi ml hydrochlor acid connect flask apparatu turn cock tube connect mobil tube fill colour sulphur acid connect graduat tube bring level liquid mark turn cock order connect tube check level run hydrochlor acid slowli portion sampl tilt flask make pressur equal lower tube shake flask releas carbon dioxid stop complet restor pressur bring liquid back level tube minut volum ga constant read carri control test condit calcium carbon calcul result content gramm carbon express calcium carbon percentag sampl calcul formula gt pic file quot quot gt ml releas portion sampl ml releas caco weight gramm portion sampl observ portion sampl weigh place ml distil water flask mix begin test volum water control test apparatu volum scheibler dietrich apparatu portion sampl control substanc calcul result adapt gt pic file quot quot gt determin crude ash purpos scope method make determin crude ash content feed stuff principl sampl ash ºc residu weigh reagent solut ammonium nitrat apparatu hot plate electr muffl furnac thermostat crucibl ash made platinum alloi platinum gold pt au rectangular mm circular diamet mm height mm procedur weigh nearest mg approxim sampl case product tendenc swell place crucibl ash calcin tare place crucibl hot plate heat gradual substanc carbonis put crucibl muffl furnac set ºc ºc temperatur white light grei reddish ash obtain appear free carbonac particl place crucibl desicc leav cool weigh immedi calcul result calcul weight residu deduct tare express result percentag sampl observ ash substanc difficult ash subject initi ash hour cool drop solut ammonium nitrat ad carefulli avoid dispers ash format lump continu calcin dry oven repeat oper ash complet case substanc resist treatment proce ash hour place ash warm water filter small ash free filter ash filter content origin crucibl place filtrat cool crucibl evapor dry ash weigh case oil fat weigh accur sampl approxim suitabl size crucibl carbonis set light substanc strip ash free filter paper combust moisten water dry ash determin ash insolubl hydrocholor acid purpos scope method make determin level feed stuff miner substanc insolubl hydrochlor acid method depend natur sampl method applic straight organ feed stuff compound feed stuff method applic miner compound mixtur compound feed stuff content substanc insolubl hydrochlor acid determin method greater principl method sampl ash ash boil hydrochlor acid insolubl residu filter weigh method sampl treat hydrochlor acid solut filter residu ash ash obtain treat accord method reagent hydrochlor acid solut trichloroacet acid solut trichloroacet acid apparatu hot plate electr muffl furnac thermostat crucibl ash made platinum alloi platinum gold pt au rectangular mm circular diamet mm height mm procedur method ash sampl method determin crude ash ash obtain analysi place ash ml beaker ml hydrochlor acid bring slowli boil boil gentli fifteen minut filter warm solut ash free filter paper wash residu warm water acid reaction longer visibl dry filter residu ash tare crucibl temperatur ºc ºc cool desicc weigh method weigh sampl nearest mg place ml beaker add ml water ml hydrochlor acid success mix wait effervesc ceas add ml hydrochlor acid wait releas ga ceas place beaker boil water bath thirti minut longer order hydrolys starch present filter warm ash free filter wash filter ml warm water observ place filter residu crucibl ash dry ash temperatur ºc ºc place ash ml beaker ml hydrochlor acid continu subparagraph calcul result calcul weight residu deduct tare express result percentag sampl observ filtrat prove difficult recomm analysi replac ml hydrochlor acid ml trichloroacet acid wash filter warm solut trichloroacet acid determin chlorin chlorid purpos scope method make determin amount chlorin chlorid solubl water convention express sodium chlorid applic feed stuff principl chlorid dissolv water product organ matter clarifi solut slightli acidifi nitric acid chlorid precipit form silver chlorid mean solut silver nitrat excess silver nitrat titrat solut ammonium thiocyan volhard method reagent solut ammonium thiocyan solut silver nitrat satur solut ammonium ferric sulphat nitric acid diethyl ether aceton carrez solut dissolv water zinc acet zn ch coo glacial acet acid make ml water carrez ii solut dissolv water potassium ferrocynaid fe cn make ml water activ carbon free chlorid absorb apparatu mixer tumbler approxim rpm procedur prepar solut natur sampl prepar solut shown time carri blank test omit sampl analys sampl free organ matter weigh nearest mg sampl chlorin form chlorid place ml water ml volumetr flask approxim ºc mix thirti minut tumbler bring volum homogenis filter sampl organ matter exclud product list weigh approxim sampl nearest mg place activ carbon ml volumetr flask add ml water approxim ºc ml carrez solut stir add ml carrez solut ii mix thirti minut tumbler bring volum homogenis filter cook feed stuff flax cake flour product rich flax flour product rich mucilag colloid substanc dextrin starch prepar solut filter decant centrifug remov ml supernat liquid transfer ml measur flask mix aceton bring volum solvent homogenis filter titrat pipett transfer erlenmay flask ml ml filtrat assum chlorin content obtain aliquot portion mg chlorin cl dilut ml water add ml nitric acid ml satur solut ammonium ferric sulphat drop ammonium thiocyan solut transfer mean burett fill mark burett transfer silver nitrat solut excess ml obtain add ml diethyl ether shake hard coagul precipit titrat excess silver nitrat ammonium thiocyan solut reddish brown tint last minut calcul result amount chlorin express sodium chlorid present volum filtrat titrat calcul formula mg ml silver nitrat solut ad ml ammonium thiocyan solut titrat blank test silver nitrat solut consum deduct volum observ titrat carri potentiometri case product rich oil fat de fat diethyl ether light petroleum case fish meal titrat carri mohr method determin mustard oil purpos scope method make determin amount mustard oil contain cake made brassica sinapi speci compound feed stuff cake made speci express allyl isothiocyan steam entrain principl sampl suspend water mustard oil releas action enzym entrain distil ethanol collect dilut ammonia solut treat warm volum silver nitrat solut cool filter excess silver nitrat titrat solut ammonium thiocyan reagent white mustard sinapi alba ethanol anti foam emuls silicon ammonia silver nitrat solut ammonium thiocyan solut nitric acid satur solut ammonium ferric sulphat apparatu flat bottom ml flask ground glass stopper distil apparatu fit condens equip prevent entrain droplet procedur weigh sampl nearest mg place ml flat bottom flask add fine ground white mustard enzym sourc ml water ºc stopper flask ºc approxim hour shake frequent add ml ethanol drop anti foam emuls distil approxim ml collect distil ml volumetr flask ml ammonia make end condens immers liquid add ammoniac solut ml silver nitrat solut place small funnel volumetr flask heat mixtur boil water bath hour leav cool bring volum water stir filter remov ml clear filtrat add ml nitric acid approxim ml ammonium ferric sulphat solut titrat excess silver nitrat return ammonium thiocyan solut carri blank test appli procedur fine ground white mustard omit sampl analysi calcul result subtract volum silver nitrat solut consum blank test consum sampl solut obtain number ml silver nitrat solut consum mustard oil sampl ml agno correspond mg allyl isothiocyan express result percentag sampl determin lactos purpos scope method make determin level lactos feed stuff lactos principl sugar dissolv water solut subject ferment yeast saccharomyc cerevisia leav lactos intact clarif filtrat lactos content filtrat determin luff schoorl method reagent suspens saccharomyc cerevisia suspend fresh yeast ml water suspens maximum period week refriger carrez solut dissolv water zinc acet zn ch coo glacial acet acid make ml water carrez solut ii dissolv water potassium ferrocyanid fe cn make ml water luff schoorl reagent stir carefulli pour citric acid solut sodium carbon solut add copper sulphat solut make litr water leav settl overnight filter check normal reagent obtain cu na solut ph approxim copper sulphat solut dissolv copper sulphat cu free iron ml water citric acid solut dissolv citric acid ml water sodium carbon solut dissolv anhydr sodium carbon approxim ml warm water leav cool granul pumic stone boil hydrochlor acid wash water dri solut sodium iodid sulphur acid solut sodium thiosulph starch solut add mixtur solubl starch ml water litr boil water boil minut leav cool add mg mercur iodid preserv apparatu water bath thermostat set ºc procedur weigh sampl nearest mg place portion sampl ml volumetr flask add ml water place flask boil water bath thirti minut cool approxim ºc add ml yeast suspens homogenis leav flask stand hour water bath temperatur ºc cool approxim ºc add ml carrez solut stir thirti second add ml carrez solut ii stir thirti second make ml water mix filter pipett remov amount filtrat exce ml prefer mg lactos transfer ml erlenmey flask make ml water carri blank test ml yeast suspens determin lactos content luff schoorl add ml luff schoorl reagent granul pumic stone stir hand heat free flame medium height bring liquid boil approxim minut place erlenmey immedi asbesto coat wire gauz hole approxim cm diamet flame lit flame regul base erlenmey heat fit reflux condens erlenmey flask boil ten minut cool immedi cold water approxim minut titrat add ml potassium iodid solut immedi carefulli risk abund foam add ml sulphur acid titrat sodium thiosulph solut dull yellow colour appear add starch indic complet titrat carri titrat accur measur mixtur ml luff schoorl reagent ml water ad ml potassium iodid solut ml sulphur acid boil calcul result attach tabl establish amount lactos mg correspond differ result titrat express ml sodium thiosulph express result part anhydr lactos percentag sampl observ product ferment sugar ml yeast suspens gt pic file quot quot gt determin potassium purpos scope method make determin level potassium feed stuff principl sampl ash ash dissolv hydrochlor acid sodium content solut determin flame photometri presenc caesium chlorid aluminium nitrat addit substanc larg elimin interfer disturb element reagent hydrochlor acid caesium chlorid aluminium nitrat gener purpos reagent potassium chlorid anhydr load agent dissolv water caesium chlorid aluminium nitrat make litr water homogenis store plastic bottl standard solut potassium dissolv water potassium chlorid add ml hydrochlor acid make litr water homogenis store plastic bottl ml solut mg potassium apparatu platinum silica porcelain crucibl ash provid lid electr muffl furnac thermostat flame photomet procedur analysi sampl gener rule weigh sampl nearest mg place crucibl ash ºc hour cool transfer ash quantit ml graduat flask ml water ml hydrochlor acid releas carbon dioxid ceas heat solut temperatur ºc hour stir occasion cool room temperatur make mark water shake filter transfer ml graduat flask aliquot part filtrat maximum mg potassium add ml load agent make mark water homogenis case higher level potassium dilut solut analys suitabl proport ad load agent tabl guid sampl gt pic file quot quot gt measur flame photometri wavelength nm calcul result mean calibr curv calibr curv place ml standard solut ml graduat flask make mark water homogenis place ml graduat flask ml solut amount potassium mg complet seri blank flask standard solut add ml load agent flask make mark water homogenis carri measur calibr curv gener linear potassium concentr mg ml solut calcul result express result percentag sampl observ add load agent order elimin interfer disturb element determin sodium purpos scope method make determin level sodium feed stuff principl sampl ash ash dissolv hydrochlor acid sodium content solut determin flame photometri presenc caesium chlorid aluminium nitrat addit substanc larg elimin interfer disturb element reagent hydrochlor acid caesium chlorid aluminium nitrat gener purpos reagent sodium chlorid anhydr load agent dissolv water caesium chlorid aluminium nitrat make litr water homogenis store plastic bottl standard solut sodium dissolv water sodium chlorid add ml hydrochlor acid make litr water homogenis store plastic bottl ml solut mg sodium apparatu platinum silica porcelain crucibl ash provid lid electr muffl furnac thermostat flame photomet procedur analysi sampl gener rule weigh sampl nearest mg place crucibl ash ºc hour avoid overh ignit cool transfer ash quantit ml graduat flask ml water ml hydrochlor acid releas carbon dioxid ceas heat solut temperatur ºc hour stir occasion cool room temperatur make mark water shake filter transfer ml graduat flask aliquot part filtrat maximum mg sodium add ml load agent make mark water homogenis case higher level sodium dilut solut analys suitabl proport ad load agent tabl guid sampl gt pic file quot quot gt measur flame photometri wavelength nm calcul result mean calibr curv calibr curv place ml standard solut ml graduat flask make mark water homogenis place ml graduat flask ml solut amount sodium mg complet seri blank flask standard solut add ml load agent flask make mark water homogenis carri measur calibr curv gener linear sodium concentr mg ml solut calcul result express result percentag sampl observ product sodium prefer ash substanc hour crucibl lid cool add water bring ash suspens mean platinum wire dry ash hour crucibl lid sampl consist sole miner substanc dissolv prior ash determin sugar purpos scope method make determin amount reduc sugar total sugar invers express glucos sucros convert factor applic compound feed stuff special method provid feed stuff lactos measur separ account calcul result principl sugar extract dilut ethanol solut clarifi carrez solut ii elimin ethanol quantiti invers determin luff schoorl method reagent ethanol ºc neutralis phenolphthalein carrez solut dissolv water zinc acet zn ch coo glacial acet acid make ml water carrez solut ii dissolv water potassium ferrocyanid fe cn make ml water solut methyl orang hydrochlor acid hydrochlor acid sodium hydroxid solut luff schoorl reagent stir carefulli pour citric acid solut sodium carbon solut add copper sulphat solut make litr water leav settl overnight filter check normal reagent obtain cu na solut ph approxim copper sulphat solut dissolv copper sulphat cu free iron ml water citric acid solut dissolv citric acid ml water sodium carbon solut dissolv anhydr sodium carbon approxim ml warm water leav cool sodium thiosulph solut starch solut add mixtur solubl starch ml water litr boil water boil minut leav cool add mg mercur iodid preserv sulphur acid solut potassium iodid granul pumic stone boil hydrochlor acid wash water dri methylbutan ol apparatu mixer tumbler approxim rpm procedur extract sampl weigh sampl nearest mg place ml volumetr flask add ml ethanol mix tumbler hour add ml carrez solut stir minut add ml carrez solut ii stir minut make volum ethanol homogenis filter remov ml filtrat evapor approxim half volum order elimin ethanol transfer evapor residu quantit ml volumetr flask warm water cool bring volum water homogenis filter solut determin amount reduc sugar invers total sugar determin reduc sugar pipett remov ml solut mg reduc sugar express glucos make ml distil water determin content reduc sugar luff schoorl method result express percentag content glucos sampl determin total sugar invers pipett ml solut transfer ml volumetr flask add drop methyl orang solut carefulli stir continu add hydrochlor acid liquid turn definit red add ml hydrochlor acid immers flask fast boil water bath thirti minut cool rapidli approxim ºc add ml sodium hydroxid solut make ml water homogenis remov ml mg reduc sugar express glucos make ml distil water determin content reduc sugar luff schoorl method result express percentag glucos sucros multipli factor titrat luff schoorl method pipett ml luff schoorl reagent transfer ml erlenmey flask add ml clarifi sugar solut add granul pumic stone heat stir hand free flame medium height bring liquid boil approxim minut place erlenmey immedi asbesto coat wire gauz hole approxim cm diamet flame lit flame regul base erlenmey heat fit reflux condens erlenmey flask boil ten minut cool immedi cold water approxim minut titrat add ml potassium iodid solut immedi carefulli risk abund foam add ml sulphur acid titrat sodium thiosulph solut dull yellow colour appear add starch indic complet titrat carri titrat accur measur mixtur ml luff schoorl reagent ml water ad ml potassium iodid solut ml sulphur acid boil calcul result tabl establish amount glucos mg correspond differ valu titrat express mg sodium thiosulph express result percentag sampl special procedur case feed stuff rich molass feed stuff homogen weigh place ml water litr volumetr flask mix hour tumbler clarifi carrez ii reagent time time quantiti reagent bring volum ethanol homogenis filter elimin ethanol dextrinis starch bring volum distil water case molass straight feed stuff rich sugar starch free carob dri beetroot cossett weigh place ml volumetr flask add ml distil water mix tumbler hour clarifi carrez ii reagent bring volum cold water homogenis filter order determin amount total sugar continu observ order prevent foam advis add irrespect volum approxim ml methylbutan ol boil luff schoorl reagent differ content total sugar invers express glucos content reduc sugar express glucos multipli percentag content sucros order determin content reduc sugar exclud lactos method adopt approxim calcul multipli lactos content establish method analysi subtact result obtain content reduc sugar accur calcul reduc sugar exclud lactos sampl final determin analys carri part solut obtain part solut obtain determin lactos method laid purpos ferment type sugar clarifi case amount sugar present determin luff schoorl method calcul mg glucos valu subtract differ express percentag sampl volum correspond determin sampl mg case ml sodium thiosulph solut mg glucos consum ml mg glucos differ mg glucos content reduc sugar exclud lactos calcul glucos gt pic file quot quot gt gt pic file quot quot gt determin theobromin purpos scope method make determin amount theobromin product process cocoa bean principl theobromin extract chloroform extract evapor dry dissolv water treat specif quantiti silver nitrat solut nitric acid liber titrat solut sodium hydroxid reagent chloroform ammonia sodium sulphat anhydr solut sodium hydroxid solut silver nitrat ethanol solut phenol red light petroleum ºc apparatu flat bottom ml flask ground glass stopper procedur weigh nearest mg sampl mg theobromin place ml flat bottom flask ground glass stopper add ml chloroform ml ammonia stopper flask shake vigor minut add anhydr sodium sulphat shake leav settl dai filter ml erlenmey wash residu ml chloroform distil solvent elimin trace boil water bath redissolv extract ml water bring boil cool neutralis sodium hydroxid solut ml phenol red solut add ml silver nitrat solut titrat nitric acid liber solut sodium hydroxid indic colour ph calcul result ml naoh mg theobromin express result percentag sampl observ product crude fatti matter defat hour extract light petroleum ºc determin urea purpos scope method make determin level urea feed stuff principl sampl suspend water clarifi agent suspens filter urea content filtrat determin addit dimethylaminobenzaldehyd dmab measur optic densiti wavelength nm reagent solut dimethylaminobenzaldehyd dissolv dmab ml ethanol add ml hydrochlor acid reagent maximum period week carrez solut dissolv water zinc acet ch coo glacial acet acid make ml water carrez solut ii dissolv water potassium ferrocyanid fe cn make ml water activ carbon absorb urea check solut urea apparatu mixer tumbler approxim rpm test tube mm ground glass stopper spectrophotomet procedur analysi sampl weigh sampl nearest mg place activ carbon ml volumetr flask add ml water ml carrez solut ii mix thirti minut tumbler make volum water shake filter remov ml transpar colourless filtrat place test tube ground glass stopper add ml dmab solut mix place tube hot water bath ºc fifteen minut measur optic densiti sampl solut spectrophotomet nm compar blank test solut reagent calibr curv remov volum ml urea solut place ml volumetr flask make volum water remov ml solut add ml dmab solut homogenis measur optic densiti shown comparison control solut ml dmab ml water free urea plot calibr curv calcul result determin amount urea sampl calibr curv express result percentag sampl observ case content urea exceed reduc sampl dilut origin solut mg urea ml case low content urea increas sampl long filtrat remain transpar colourless sampl simpl nitrogen compound amino acid optic densiti measur nm determin lupin alkaloid purpos scope method make determin level alkaloid lupin seed principl alkaloid dissolv mixtur diethyl ether chloroform extract hydrochlor acid alkaloid precipit silicotungst acid precipit ash residu weigh reagent diethyl ether chloroform sodium hydroxid solut hydrochlor acid sodium chlorid solut silicotungst acid wo apparatu mechan stirrer platinum silica porcelain crucibl ash electr muffl furnac procedur weigh sampl nearest mg place contain capac approxim ml fit ground glass stopper separ funnel add ml diethyl ether ml chloroform graduat pipett ml sodium hydroxid solut shake vigor prevent agglomer shake time leav stand overnight supernat liquid complet clear add drop water filter etherchloroform layer collect ml filtrat ml volumetr flask transfer quantit ml separ funnel ml diethyl ether extract time success ml hydrochlor acid leav decant collect acid extract extract collect acid extract ml beaker elimin trace ether chloroform warm gentli add approxim sodium chlorid leav cool precipit alkaloid silicotungst acid solut stir mechan stirrer thirti minut leav settl overnight filter ash free filter wash precipit success ml ml hydrochlor acid place filter precipit crucibl ash ºc leav cool weigh calcul result alkaloid content sampl obtain multipli weight ash factor express result percentag sampl estim ureas activ product deriv soya purpos scope test make estim ureas activ product deriv soya show product cook suffici length time principl ureas activ estim amount ammoniac nitrogen liber product minut ºc solut urea reagent hydrochlor acid solut sodium hydroxid load agent phosphat ml disodium phosphat na hpo monopotassium phosphat kh po freshli prepar urea load agent urea ml load agent ph apparatu potentiometr titrat apparatu high sensit ph meter ph magnet stirrer water bath fit thermostat set ºc test tube ground glass stopper mm procedur crush sampl coffe mill pass siev mesh mm weigh crush sampl nearest mg place test tube ground glass stopper add ml urea load agent stopper immedi shake vigor place tube water bath set ºc shake vigor place tube water bath set ºc thirti minut immedi add ml hydrochlor acid cool rapidli ºc transfer content tube quantit titrat vessel rins ml water glass electrod titrat immedi rapidli ph sodium hydroxid solut electrometri carri blank test quickli place sampl weigh nearest mg test tube ground glass stopper add ml hydrochlor acid ml urea load agent cool tube immedi ic water leav thirti minut condit transfer content tube titrat vessel sodium hydroxid solut ph calcul ureas activ calcul formula gt pic file quot quot gt ml sodium hydroxid solut consum sampl ml sodium hydroxid solut consum blank test weight sampl observ method suitabl ureas activ mg min ºc activ product size sampl reduc mg product crude fatti substanc defat cold