美国药典USP32测试溶液配制

TESTSOLUTIONS(TS)Certainofthefollowingtestsolutionsareintendedforuseasacid-baseindicatorsinvolumetricanalyses.Suchsolutionsshouldbesoadjustedthatwhen0.15mLoftheindicatorsolutionisaddedto25mLofcarbondioxide-freewater,0.25mLof0.02Nacidoralkali,respectively,willproducethecharacteristiccolorchange.SimilarsolutionsareintendedforuseinpHmeasurement.Wherenospecialdirectionsfortheirpreparationaregiven,thesamesolutionissuitableforbothpurposes.Whereitisdirectedthatavolumetricsolutionbeusedasthetestsolution,standardizationofthesolutionusedasTSisnotrequired.Ingeneral,thedirectivetoprepareasolution“fresh”indicatesthatthesolutionisoflimitedstabilityandmustbepreparedonthedayofuse.ForthepreparationofTestSolutions,usereagentsofthequalitydescribedunderReagents.AcetateBufferTS—Dissolve320gofammoniumacetatein500mLofwater,add5mLofglacialaceticacid,dilutewithwaterto1000.0mL,andmix.ThissolutionhasapHbetween5.9and6.0.AceticAcid,Glacial,TS—DeterminethewatercontentofaspecimenofglacialaceticacidbytheTitrimetricMethod(seeWaterDetermination921).Iftheacidcontainsmorethan0.05%ofwater,addafewmLofaceticanhydride,mix,allowtostandovernight,andagaindeterminethewatercontent.Iftheacidcontainslessthan0.02%ofwater,addsufficientwatertomakethefinalconcentrationbetween0.02%and0.05%,mix,allowtostandovernight,andagaindeterminethewatercontent.Repeattheadjustmentwithaceticanhydrideorwater,asnecessary,untiltheresultingsolutionshowsawatercontentbetween0.02%and0.05%.AceticAcid,Strong,TS—Add300.0mLofglacialaceticacid,anddilutewithwaterto1000mL.Thissolutioncontainsabout30%(v/v)ofCH3COOHandhasaconcentrationofabout5N.AceticAcid–AmmoniumAcetateBufferTS—Dissolve77.1gofammoniumacetateinwater,add57mLofglacialaceticacid,anddilutewithwaterto1000mL.Acetone,Buffered,TS—Dissolve8.15gofsodiumacetateand42gofsodiumchlorideinabout100mLofwater,andadd68mLof0.1Nhydrochloricacidand150mLofacetone.Mix,anddilutewithwaterto500mL.AcidFerricChlorideTS—Mix60mLofglacialaceticacidwith5mLofsulfuricacid,add1mLofferricchlorideTS,mix,andcool.AcidFerrousSulfateTS—SeeFerrousSulfate,Acid,TS.AcidStannousChlorideTS—SeeStannousChloride,Acid,TS.AcidStannousChlorideTS,Stronger—SeeStannousChloride,Acid,TS.AlbumenTS—Carefullyseparatethewhitefromtheyolkofastrictlyfreshhen'segg.Shakethewhitewith100mLofwateruntilmixedandallbutthechalazahasundergonesolution;thenfilter.Preparethesolutionfresh.Alcohol–PhenolTS—Dissolve780mgofphenolinalcoholtomake100mL.AlcoholicAmmoniaTS—SeeAmmoniaTS,Alcoholic.AlcoholicMercuricBromideTS—SeeMercuricBromideTS,Alcoholic.AlcoholicPotassiumHydroxideTS—SeePotassiumHydroxideTS,Alcoholic.AlkalineCupricCitrateTS—SeeCupricCitrateTS,Alkaline.AlkalineCupricCitrateTS2—SeeCupricCitrateTS2,Alkaline.AlkalineCupricIodideTS—SeeCupricIodideTS,Alkaline.AlkalineCupricTartrateTS(Fehling'sSolution)—SeeCupricTartrateTS,Alkaline.AlkalineMercuric–PotassiumIodideTS—SeeMercuric–PotassiumIodideTS,Alkaline.AlkalinePicrateTS—SeePicrateTS,Alkaline.AlkalineSodiumHydrosulfiteTS—SeeSodiumHydrosulfiteTS,Alkaline.AmaranthTS—Dissolve20mgofamaranthin10mLofwater.AminonaphtholsulfonicAcidTS—Accuratelyweigh5gofsodiumsulfite,94.3gofsodiumbisulfite,and700mgof1,2,4-aminonaphtholsulfonicacid,andmix.PrepareaminonaphtholsulfonicacidTSfreshonthedayofusebydissolving1.5gofthedrymixturein10mLofwater.Ammonia–AmmoniumChlorideBufferTS—Dissolve67.5gofammoniumchlorideinwater,add570mLofammoniumhydroxide,anddilutewithwaterto1000mL.Ammonia–CyanideTS—Dissolve2gofpotassiumcyanidein15mLofammoniumhydroxide,anddilutewithwaterto100mL.AmmoniaTS—Itcontainsbetween9.5%and10.5%ofNH3.Preparebydiluting350mLofAmmoniaWater,Stronger(seeinthesection,Reagents)withwatertomake1000mL.AmmoniaTS,Alcoholic—Asolutionofammoniagasinalcohol.Clear,colorlessliquidhavingastrongodorofammonia.Specificgravity:about0.80.Itcontainsbetween9%and11%ofNH3.Storeitinalkali-resistantcontainers,inacoldplace.AmmoniaTS,Stronger—UseAmmoniaWater,Stronger(seeinthesectionReagents).AmmoniacalPotassiumFerricyanideTS—Dissolve2gofpotassiumferricyanidein75mLofwater,add25mLofammoniumhydroxide,andmix.AmmoniatedCupricOxideTS—SeeCupricOxide,Ammoniated,TS.AmmoniumAcetateTS—Dissolve10gofammoniumacetateinwatertomake100mL.AmmoniumCarbonateTS—Dissolve20gofammoniumcarbonateand20mLofammoniaTSinwatertomake100mL.AmmoniumChlorideTS—Dissolve10.5gofammoniumchlorideinwatertomake100mL.AmmoniumChloride–AmmoniumHydroxideTS—Mixequalvolumesofwaterandammoniumhydroxide,andsaturatewithammoniumchloride.AmmoniumMolybdateTS—Dissolve6.5goffinelypowderedmolybdicacidinamixtureof14mLofwaterand14.5mLofammoniumhydroxide.Coolthesolution,andadditslowly,withstirring,toawell-cooledmixtureof32mLofnitricacidand40mLofwater.Allowtostandfor48hours,andfilterthroughafine-porosity,sintered-glasscrucible.Thissolutiondeterioratesuponstandingandisunsuitableforuseif,upontheadditionof2mLofdibasicsodiumphosphateTSto5mLofthesolution,anabundantyellowprecipitatedoesnotformatonceorafterslightwarming.Storeitinthedark.Ifaprecipitateformsduringstorage,useonlytheclearsupernatant.AmmoniumOxalateTS—Dissolve3.5gofammoniumoxalateinwatertomake100mL.AmmoniumPhosphate,Dibasic,TS(AmmoniumPhosphateTS)—Dissolve13gofdibasicammoniumphosphateinwatertomake100mL.AmmoniumPolysulfideTS—Yellowliquid,madebysaturatingammoniumsulfideTSwithsulfur.AmmoniumPyrrolidinedithiocarbamate,Saturated,TS—Addabout10gofammoniumpyrrolidinedithiocarbamatetoa1000-mLvolumetricflask,anddilutewithwatertovolume.AmmoniumReineckateTS—Shakeabout500mgofammoniumreineckatewith20mLofwaterfrequentlyduring1hour,andfilter.Usewithin2days.AmmoniumSulfideTS—SaturateammoniaTSwithhydrogensulfidebybubblinghydrogensulfidegasthroughthesolutionfor1minute.Thissolutionmustbefreshlyprepared.ThesolutionisnotrenderedturbideitherbymagnesiumsulfateTSorbycalciumchlorideTS(carbonate).Thissolutionisunstableforuseifanabundantprecipitateofsulfurispresent.Residueonignition:notmorethan0.05%.AmmoniumThiocyanateTS—Dissolve8gofammoniumthiocyanateinwatertomake100mL.AmmoniumVanadateTS—Dissolve2.5gofammoniumvanadatein500mLofboilingwater,cool,andadd20mLofnitricacid.Mix,cool,andaddwatertomake1L.Storeinpolyethylenecontainers.AnthroneTS—Within12hoursofuse,rapidlydissolve35mgofanthroneinahotmixtureof35mLofwaterand65mLofsulfuricacid.Immediatelycoolinanicebathtoroomtemperature,andfilterthroughglasswool.Allowthesolutiontostandatroomtemperaturefor30minutesbeforeuse.AntimonyTrichlorideTS—Dissolve20gofantimonytrichlorideinchloroformtomake100mL.Filterifnecessary.BariumChlorideTS—Dissolve12gofbariumchlorideinwatertomake100mL.BariumHydroxideTS—Asaturatedsolutionofbariumhydroxideinrecentlyboiledwater.Preparethesolutionfresh.BariumNitrateTS—Dissolve6.5gofbariumnitrateinwatertomake100mL.BetanaphtholTS—See2-NaphtholTS.BiuretReagentTS—Dissolve1.5gofcupricsulfateand6.0gofpotassiumsodiumtartratein500mLofwaterina1000-mLvolumetricflask.Add300mLofcarbonate-freesodiumhydroxidesolution(1in10),dilutewithcarbonate-freesodiumhydroxidesolution(1in10)to1000mL,andmix.BlueTetrazoliumTS—Dissolve500mgofbluetetrazoliuminalcoholtomake100mL.BrilliantBlueGTS—Transfer25mgofbrilliantblueGtoa100-mLvolumetricflask,add12.5mLofalcoholand25mLofphosphoricacid,dilutewithwatertovolume,andmix.BromineTS(BromineWater)—Asaturatedsolutionofbromine,preparedbyagitating2to3mLofbrominewith100mLofcoldwaterinaglass-stopperedbottle,thestopperofwhichshouldbelubricatedwithpetrolatum.Storeitinacoldplace,protectedfromlight.Bromine–SodiumAcetateTS—Dissolve100gofsodiumacetatein1000mLofglacialaceticacid,add50mLofbromine,andmix.p-BromoanilineTS—Add8gofp-bromoanilinetoamixtureof380mLofthiourea-saturatedglacialaceticacid,10mLofsodiumchloridesolution(1in5),5mLofoxalicacidsolution(1in20),and5mLofdibasicsodiumphosphatesolution(1in10)inalow-actinicglassbottle.Mix,andallowtostandovernightbeforeusing.Protectfromlight,andusewithin7days.BromocresolBlueTS—UseBromocresolGreenTS.BromocresolGreenTS—Dissolve50mgofbromocresolgreenin100mLofalcohol,andfilterifnecessary.BromocresolGreen–MethylRedTS—Dissolve0.15gofbromocresolgreenand0.1gofmethylredin180mLofalcohol,anddilutewithwaterto200mL.BromocresolPurpleTS—Dissolve250mgofbromocresolpurplein20mLof0.05Nsodiumhydroxide,anddilutewithwaterto250mL.BromophenolBlueTS—Dissolve100mgofbromophenolbluein100mLofdilutedalcohol,andfilterifnecessary.BromothymolBlueTS—Dissolve100mgofbromothymolbluein100mLofdilutedalcohol,andfilterifnecessary.BufferedAcetoneTS—SeeAcetone,Buffered,TS.CalciumChlorideTS—Dissolve7.5gofcalciumchlorideinwatertomake100mL.CalciumHydroxideTS—UseCalciumHydroxideTopicalSolution(USPmonograph).CalciumSulfateTS—Asaturatedsolutionofcalciumsulfateinwater.CericAmmoniumNitrateTS—Dissolve6.25gofcericammoniumnitratein10mLof0.25Nnitricacid.Usewithin3days.ChloralHydrateTS—Dissolve50gofchloralhydrateinamixtureof15mLofwaterand10mLofglycerin.ChlorineTS(ChlorineWater)—Asaturatedsolutionofchlorineinwater.Placethesolutioninsmall,completelyfilled,light-resistantcontainers.ChlorineTS,evenwhenkeptfromlightandair,isapttodeteriorate.Storeitinacold,darkplace.Forfullstrength,preparethissolutionfresh.ChromotropicAcidTS—Dissolve50mgofchromotropicacidoritsdisodiumsaltin100mLof75%sulfuricacid,whichmaybemadebycautiouslyadding75mLofsulfuricacidto33.3mLofwater.Cobalt–UranylAcetateTS—Dissolve,withwarming,40gofuranylacetateinamixtureof30gofglacialaceticacidandsufficientwatertomake500mL.Similarly,prepareasolutioncontaining200gofcobaltousacetateinamixtureof30gofglacialaceticacidandsufficientwatertomake500mL.Mixthetwosolutionswhilestillwarm,andcoolto20.Maintainthetemperatureat20forabout2hourstoseparatetheexcesssaltsfromsolution,andthenpassthroughadryfilter.CobaltousChlorideTS—Dissolve2gofcobaltouschloridein1mLofhydrochloricacidandsufficientwatertomake100mL.CongoRedTS—Dissolve500mgofcongoredinamixtureof10mLofalcoholand90mLofwater.m-CresolPurpleTS—Dissolve0.10gofmetacresolpurplein13mLof0.01Nsodiumhydroxide,dilutewithwaterto100mL,andmix.CresolRedTS—Triturate100mgofcresolredinamortarwith26.2mLof0.01Nsodiumhydroxideuntilsolutioniscomplete,thendilutethesolutionwithwaterto250mL.CresolRed–ThymolBlueTS—Add15mLofthymolblueTSto5mLofcresolredTS,andmix.CrystalVioletTS—Dissolve100mgofcrystalvioletin10mLofglacialaceticacid.CupricAcetateTS—Dissolve100mgofcupricacetateinabout5mLofwatertowhichafewdropsofaceticacidhavebeenadded.Diluteto100mL,andfilter,ifnecessary.CupricAcetateTS,Stronger(Barfoed'sReagent)—Dissolve13.3gofcupricacetateinamixtureof195mLofwaterand5mLofaceticacid.Cupric-AmmoniumSulfateTS—TocupricsulfateTSaddammoniaTS,dropwise,untiltheprecipitateinitiallyformedisnearlybutnotcompletelydissolved.Allowtosettle,anddecanttheclearsolution.Preparethissolutionfresh.CupricCitrateTS—Dissolve25gofcupricsulfate,50gofcitricacid,and144gofanhydroussodiumcarbonateinwater,anddilutewithwaterto1000mL.CupricCitrateTS,Alkaline—Withtheaidofheat,dissolve173gofdihydratedsodiumcitrateand117gofmonohydratedsodiumcarbonateinabout700mLofwater,andfilterthroughpaper,ifnecessary,toobtainaclearsolution.Inaseparatecontainerdissolve17.3gofcupricsulfateinabout100mLofwater,andslowlyaddthissolution,withconstantstirring,tothefirstsolution.Coolthemixture,addwatertomake1000mL,andmix.CupricCitrateTS2,Alkaline—Withtheaidofheat,dissolveabout173gofsodiumcitratedihydrateand117gofsodiumcarbonatemonohydrateinabout700mLofwater,andfilter.Inasecondflask,dissolveabout27.06gofcupricsulfate(Cu2O4·5H2O)inabout100mLofwater.Slowlycombinethetwosolutionswhilestirring,anddilutewithwaterto1000mL.CupricIodideTS,Alkaline—Dissolve7.5gofcupricsulfate(CuSO4·5H2O)inabout100mLofwater.Inaseparatecontainerdissolve25gofanhydroussodiumcarbonate,20gofsodiumbicarbonate,and25gofpotassiumsodiumtartrateinabout600mLofwater.Withconstantstirring,addthecupricsulfatesolutiontothebottomofthealkalinetartratesolutionbymeansofafunnelthattouchesthebottomofthecontainer.Add1.5gofpotassiumiodide,200gofanhydroussodiumsulfate,50to150mLof0.02Mpotassiumiodate,andsufficientwatertomake1000mL.CupricOxide,Ammoniated,TS(Schweitzer'sReagent)—Dissolve10gofcupricsulfatein100mLofwater,addsufficientsodiumhydroxidesolution(1in5)toprecipitatethecopperhydroxide,collectthelatteronafilter,andwashfreefromsulfatewithcoldwater.Dissolvetheprecipitate,whichmustbekeptwetduringtheentireprocess,intheminimumquantityofammoniaTSnecessaryforcompletesolution.CupricSulfateTS—Dissolve12.5gofcupricsulfateinwatertomake100mL.CupricTartrateTS,Alkaline(Fehling'sSolution)—TheCopperSolution(A)—Dissolve34.66gofcarefullyselected,smallcrystalsofcupricsulfate,showingnotraceofefflorescenceofadheringmoisture,inwatertomake500mL.Storethissolutioninsmall,tightcontainers.TheAlkalineTartrateSolution(B)—Dissolve173gofcrystallizedpotassiumsodiumtartrateand50gofsodiumhydroxideinwatertomake500mL.Storethissolutioninsmall,alkali-resistantcontainers.Foruse,mixexactlyequalvolumesofSolutionsAandBatthetimerequired.Delafield'sHematoxylinTS—Prepare400mLofasaturatedsolutionofammoniumalum(SolutionA).Dissolve4gofhematoxylinin25mLofalcohol,mixitwithSolutionA,andallowittostandfor4daysinaflaskclosedwithapledgetofpurifiedcottonandexposedtolightandair(SolutionB).ThenfilterSolutionB,andaddtoitaSolutionCconsistingofamixtureof100mLofglycerinand100mLofmethanol.Mix,andallowthemixturetostandinawarmplace,exposedtolight,for6weeksuntilitbecomesdark-colored.Storeintightlystopperedbottles.Foruseinstainingendocrinetissue,dilutethistestsolutionwithanequalvolumeofwater.Denigès'Reagent—SeeMercuricSulfateTS.DiazobenzenesulfonicAcidTS—Placeinabeaker1.57gofsulfanilicacid,previouslydriedat105for3hours,add80mLofwaterand10mLofdilutedhydrochloricacid,andwarmonasteambathuntildissolved.Coolto15(someofthesulfanilicacidmayseparatebutwilldissolvelater),andaddslowly,withconstantstirring,6.5mLofsodiumnitritesolution(1in10).Thendilutewithwaterto100mL.DichlorofluoresceinTS—Dissolve100mgofdichlorofluoresceinin60mLofalcohol,add2.5mLof0.1Nsodiumhydroxide,mix,anddilutewithwaterto100mL.2,7-DihydroxynaphthaleneTS—Dissolve100mgof2,7-dihydroxynaphthalenein1000mLofsulfuricacid,andallowthesolutiontostanduntiltheyellowcolordisappears.Ifthesolutionisverydark,discarditandprepareanewsolutionfromadifferentsupplyofsulfuricacid.Thissolutionisstableforapproximately1monthifstoredinadarkbottle.DiiodofluoresceinTS—Dissolve500mgofdiiodofluoresceininamixtureof75mLofalcoholand30mLofwater.DilutedLeadSubacetateTS—SeeLeadSubacetateTS,Diluted.p-DimethylaminobenzaldehydeTS—Dissolve125mgofp-dimethylaminobenzaldehydeinacooledmixtureof65mLofsulfuricacidand35mLofwater,andadd0.05mLofferricchlorideTS.Usewithin7days.DinitrophenylhydrazineTS—Carefullymix10mLofwaterand10mLofsulfuricacid,andcool.Tothemixture,containedinaglass-stopperedflask,add2gof2,4-dinitrophenylhydrazine,andshakeuntildissolved.Tothesolutionadd35mLofwater,mix,cool,andfilter.DiphenylamineTS—Dissolve1.0gofdiphenylaminein100mLofsulfuricacid.Thesolutionshouldbecolorless.DiphenylcarbazoneTS—Dissolve1gofcrystallinediphenylcarbazonein75mLofalcohol,thenaddalcoholtomake100mL.Storeinabrownbottle.DithizoneTS—Dissolve25.6mgofdithizonein100mLofalcohol.Storeinacoldplace,andusewithin2months.Dragendorff'sTS—Mix850mgofbismuthsubnitratewith40mLofwaterand10mLofglacialaceticacid(SolutionA).Dissolve8gofpotassiumiodidein20mLofwater(SolutionB).MixequalportionsofSolutionAandSolutionBtoobtainastocksolution,whichcanbestoredforseveralmonthsinadarkbottle.Mix10mLofthestocksolutionwith20mLofglacialaceticacid,anddilutewithwatertomake100mL.EdetateDisodiumTS—Dissolve1gofedetatedisodiumin950mLofwater,add50mLofalcohol,andmix.EosinYTS(adsorptionindicator)—Dissolve50mgofeosinYin10mLofwater.EriochromeBlackTS—Dissolve200mgoferiochromeblackTand2gofhydroxylaminehydrochlorideinmethanoltomake50mL.EriochromeCyanineTS—Dissolve750mgoferiochromecyanineRin200mLofwater,add25gofsodiumchloride,25gofammoniumnitrate,and2mLofnitricacid,anddilutewithwaterto1000mL.Fehling'sSolution—SeeCupricTartrateTS,Alkaline.FerricAmmoniumSulfateTS—Dissolve8gofferricammoniumsulfateinwatertomake100mL.FerricChlorideTS—Dissolve9gofferricchlorideinwatertomake100mL.FerroinTS—Dissolve0.7gofferroussulfateand1.76gofo-phenanthrolinemonohydrochloridemonohydrateinwater,anddilutewithwaterto100mL.FerrousSulfateTS—Dissolve8gofclearcrystalsofferroussulfateinabout100mLofrecentlyboiledandthoroughlycooledwater.Preparethissolutionfresh.FerrousSulfate,Acid,TS—Dissolve7gofferroussulfatecrystalsin90mLofrecentlyboiledandthoroughlycooledwater,andaddsulfuricacidtomake100mL.Preparethissolutionimmediatelypriortouse.Folin-CiocalteuPhenolTS—Intoa1500-mLflaskintroduce100gofsodiumtungstate,25gofsodiummolybdate,700mLofwater,50mLofphosphoricacid,and100mLofhydrochloricacid.Gentlyrefluxthemixtureforabout10hours,andadd150goflithiumsulfate,50mLofwater,andafewdropsofbromine.Boilthemixture,withoutthecondenser,forabout15minutes,oruntiltheexcessbromineisexpelled.Cool,dilutewithwaterto1L,andfilter:thefiltratehasnogreenishtint.Beforeuse,dilute1partofthefiltratewith1partofwater.Whenusedforproteindetermination(i.e.,Lowryassay),thisreagentmustbefurtherdiluted(1:5)withwater.SeeMethod2inTotalProteinAssayunderBiotechnology-DerivedArticles—TotalProteinAssay1057.FormaldehydeTS—UseFormaldehydeSolution(seeinthesectionReagents).Fuchsin–PyrogallolTS—Dissolve100mgofbasicfuchsinin50mLofwaterthatpreviouslyhasbeenboiledfor15minutesandallowedtocoolslightly.Cool,add2mLofasaturatedsolutionofsodiumbisulfite,mix,andallowtostandfornotlessthan3hours.Add0.9mLofhydrochloricacid,mix,andallowtostandovernight.Add100mgofpyrogallol,shakeuntilsolutioniseffected,anddilutewithwaterto100mL.Storeinanamber-coloredglassbottleinarefrigerator.Fuchsin–SulfurousAcidTS—Dissolve200mgofbasicfuchsinin120mLofhotwater,andallowthesolutiontocool.Addasolutionof2gofanhydroussodiumsulfitein20mLofwater,thenadd2mLofhydrochloricacid.Dilutethesolutionwithwaterto200mL,andallowtostandforatleast1hour.Preparethissolutionfresh.GastricFluid,Simulated,TS—Dissolve2.0gofsodiumchlorideand3.2gofpurifiedpepsin,thatisderivedfromporcinestomachmucosa,withanactivityof800to2500unitspermgofprotein,in7.0mLofhydrochloricacidandsufficientwatertomake1000mL.[note—PepsinactivityisdescribedintheFoodChemicalsCodexspecificationsunderGeneralTestsandAssays.]ThistestsolutionhasapHofabout1.2.GelatinTS(fortheassayofCorticotropinInjection)—Dissolve340gofacid-treatedprecursorgelatin(TypeA)inwatertomake1000mL.Heatthesolutioninanautoclaveat115for30minutesaftertheexhaustlinetemperaturehasreached115.Coolthesolution,andadd10gofphenoland1000mLofwater.Storeintightcontainersinarefrigerator.GlacialAceticAcidTS—SeeAceticAcid,Glacial,TS.GlucoseOxidase–ChromogenTS—Asolutioncontaining,ineachmL,0.5µmolof4-aminoantipyrine,22.0µmolofsodiump-hydroxybenzoate,notlessthan7.0unitsofglucoseoxidase,andnotlessthan0.5unitsofperoxidase,andbufferedtoapHof7.0±0.1.Suitability—WhenusedfordeterminingglucoseinInulin,ascertainthatnosignificantcolorresultsbyreactionwithfructose,andthatasuitableabsorbance-versus-concentrationslopeisobtainedwithglucose.[note—Asuitablegradeisavailable,asaconcentrate,fromWorthingtonDiagnostics,DivisionofMilliporeCorp.,www.millipore.com.]GlycerinBaseTS—To200gofglycerinaddwatertobringthetotalweightto235g.Add140mLof1Nsodiumhydroxideand50mLofwater.GoldChlorideTS—Dissolve1gofgoldchloridein35mLofwater.HydrogenPeroxideTS—UseHydrogenPeroxideTopicalSolution(USPmonograph).HydrogenSulfideTS—Asaturatedsolutionofhydrogensulfide,madebypassingH2Sintocoldwater.Storeitinsmall,darkamber-coloredbottles,fillednearlytothetop.ItisunsuitableunlessitpossessesastrongodorofH2S,andunlessitproducesatonceacopiousprecipitateofsulfurwhenaddedtoanequalvolumeofferricchlorideTS.Storeinacold,darkplace.HydroxylamineHydrochlorideTS—Dissolve3.5gofhydroxylaminehydrochloridein95mLof60%alcohol,andadd0.5mLofbromophenolbluesolution(1in1000ofalcohol)and0.5Nalcoholicpotassiumhydroxideuntilagreenishtintdevelopsinthesolution.Thenadd60%alcoholtomake100mL.8-HydroxyquinolineTS—Dissolve5gof8-hydroxyquinolineinalcoholtomake100mL.IndigoCarmineTS(SodiumIndigotindisulfonateTS)—Dissolveaquantityofsodiumindigotindisulfonate,equivalentto180mgofC16H8N2O2(SO3Na)2,inwatertomake100mL.Usewithin60days.Indophenol–AcetateTS(fortheassayofCorticotropinInjection)—To60mLofstandarddichlorophenol-indophenolsolution(seeinthesectionVolumetricSolutions)addwatertomake250mL.Addtotheresultingsolutionanequalvolumeofsodiumacetatesolutionfreshlypreparedbydissolving13.66gofanhydroussodiumacetateinwatertomake500mLandadjustingwith0.5NaceticacidtoapHof7.Storeinarefrigerator,andusewithin2weeks.IntestinalFluid,Simulated,TS—Dissolve6.8gofmonobasicpotassiumphosphatein250mLofwater,mix,andadd77mLof0.2Nsodiumhydroxideand500mLofwater.Add10.0gofpancreatin,mix,andadjusttheresultingsolutionwitheither0.2Nsodiumhydroxideor0.2NhydrochloricacidtoapHof6.8±0.1.Dilutewithwaterto1000mL.IodineTS—Use0.1NIodine(seeinthesectionVolumetricSolutions).Iodine,DilutedTS—Transfer10.0mLof0.1NiodineVStoa100-mLvolumetricflask,dilutewithwatertovolume,andmix.IodineMonochlorideTS—Dissolve10gofpotassiumiodideand6.44gofpotassiumiodatein75mLofwaterinaglass-stopperedcontainer.Add75mLofhydrochloricacidand5mLofchloroform,andadjusttoafaintiodinecolor(inthechloroform)byaddingdilutepotassiumiodideorpotassiumiodatesolution.Ifmuchiodineisliberated,useastrongersolutionofpotassiumiodatethan0.01Matfirst,makingthefinaladjustmentwiththe0.01Mpotassiumiodate.Storeinadarkplace,andreadjusttoafaintiodinecolorasnecessary.IodineandPotassiumIodideTS1—Dissolve500mgofiodineand1.5gofpotassiumiodidein25mLofwater.IodineandPotassiumIodideTS2—Dissolve12.7gofiodineand20gofpotassiumiodideinwater,anddilutewithwaterto1000.0mL.To10.0mLofthissolution,add0.6gofpotassiumiodide,anddilutewithwaterto100.0mL.Prepareimmediatelybeforeuse.IodobromideTS—Dissolve20gofiodinemonobromideinglacialaceticacidtomake1000mL.Storeinglasscontainers,protectedfromlight.IodochlorideTS—Dissolve16.5gofiodinemonochloridein1000mLofglacialaceticacid.IodoplatinateTS—Dissolve300mgofplatinicchloridein97mLofwater.Immediatelypriortouse,add3.5mLofpotassiumiodideTS,andmix.Iron–PhenolTS(Iron-KoberReagent)—Dissolve1.054gofferrousammoniumsulfatein20mLofwater,andadd1mLofsulfuricacidand1mLof30percenthydrogenperoxide.Mix,heatuntileffervescenceceases,anddilutewithwaterto50mL.To3volumesofthissolutioncontainedinavolumetricflaskaddsulfuricacid,withcooling,tomake100volumes.Purifyphenolbydistillation,discardingthefirst10%andthelast5%,collectingthedistillate,withexclusionofmoisture,inadry,taredglass-stopperedflaskofabouttwicethevolumeofthephenol.Solidifythephenolinanicebath,breakingthetopcrustwithaglassrodtoensurecompletecrystallization.Weightheflaskanditscontents,addtothephenol1.13timesitsweightoftheiron–sulfuricacidsolutionpreparedasdirected,insertthestopperintheflask,andallowtostand,withoutcoolingbutwithoccasionalmixing,untilthephenolisliquefied.Shakethemixturevigorouslyuntilmixed,allowtostandinthedarkfor16to24hours,andagainweightheflaskanditscontents.Tothemixtureadd23.5%ofitsweightofasolutionof100volumesofsulfuricacidin110volumesofwater,mix,transfertodryglass-stopperedbottles,andstoreinthedark,protectedfromatmosphericmoisture.Usewithin6months.Dispensethereagentfromasmall-boreburet,arrangedtoexcludemoisture,capableofdelivering1mLin30secondsorless,andhavingnolubricant,otherthanreagent,onitsstopcock.Wipetheburettipwithtissuebeforeeachaddition.IronSalicylateTS—Dissolve500mgofferricammoniumsulfatein250mLofwatercontaining10mLofdilutedsulfuricacid,andaddwatertomake500mL.To100mLoftheresultingsolutionadd50mLofa1.15%solutionofsodiumsalicylate,20mLofdilutedaceticacid,and80mLofa13.6%solutionofsodiumacetate,thenaddwatertomake500mL.Storeinawell-closedcontainer.Protectfromlight.Usewithin2weeks.LeadAcetateTS—Dissolve9.5gofclear,transparentcrystalsofleadacetateinrecentlyboiledwatertomake100mL.Storeinwell-stopperedbottles.LeadAcetateTS,Alcoholic—Dissolve2gofclear,transparentcrystalsofleadacetateinalcoholtomake100mL.Storeintightcontainers.LeadSubacetateTS—Triturate14gofleadmonoxidetoasmoothpastewith10mLofwater,andtransferthemixturetoabottle,usinganadditional10mLofwaterforrinsing.Dissolve22gofleadacetatein70mLofwater,andaddthesolutiontotheleadoxidemixture.Shakeitvigorouslyfor5minutes,thensetitaside,shakingitfrequently,during7days.Finallyfilter,andaddenoughrecentlyboiledwaterthroughthefiltertomake100mL.LeadSubacetateTS,Diluted—Dilute3.25mLofleadsubacetateTSwithwater,recentlyboiledandcooled,tomake100mL.Storeinsmall,well-filled,tightcontainers.LitmusTS—Digest25gofpowderedlitmuswiththreesuccessive100-mLportionsofboilingalcohol,continuingeachextractionforabout1hour.Filter,washwithalcohol,anddiscardthealcoholfiltrate.Maceratetheresiduewithabout25mLofcoldwaterfor4hours,filter,anddiscardthefiltrate.Finallydigesttheresiduewith125mLofboilingwaterfor1hour,cool,andfilter.Locke-Ringer'sSolution—SeeLocke-Ringer'sTS.Locke-Ringer'sTS(Locke-Ringer'sSolution)— SodiumChloride 9.0g PotassiumChloride 0.42g CalciumChloride 0.24g MagnesiumChloride 0.2g SodiumBicarbonate 0.5g Dextrose 0.5g Water,recentlydistilledfromahard-glassflask,asufficientquantitytomake 1000mLPreparefresheachday.Theconstituents(exceptthedextroseandthesodiumbicarbonate)maybemadeupinstocksolutionsanddilutedasneeded.MagnesiaMixtureTS—Dissolve5.5gofmagnesiumchlorideand7gofammoniumchloridein65mLofwater,add35mLofammoniaTS,setthemixtureasideforafewdaysinawell-stopperedbottle,andfilter.Ifthesolutionisnotperfectlyclear,filteritbeforeusing.MagnesiumSulfateTS—Dissolve12gofcrystalsofmagnesiumsulfate,selectedforfreedomfromefflorescence,inwatertomake100mL.MalachiteGreenTS—Dissolve1gofmalachitegreenoxalatein100mLofglacialaceticacid.Mallory'sStain—Dissolve500mgofwater-solubleanilineblue,2goforangeG,and2gofoxalicacidin100mLofwater.Mayer'sReagent—SeeMercuric–PotassiumIodideTS.MercuricAcetateTS—Dissolve6.0gofmercuricacetateinglacialaceticacidtomake100mL.Storeintightcontainers,protectedfromdirectsunlight.Mercuric–AmmoniumThiocyanateTS—Dissolve30gofammoniumthiocyanateand27gofmercuricchlorideinwatertomake1000mL.MercuricBromideTS,Alcoholic—Dissolve5gofmercuricbromidein100mLofalcohol,employinggentleheattofacilitatesolution.Storeinglasscontainers,protectedfromlight.MercuricChlorideTS—Dissolve6.5gofmercuricchlorideinwatertomake100mL.MercuricIodideTS(Valser'sReagent)—Slowlyaddpotassiumiodidesolution(1in10)toredmercuriciodideuntilalmostallofthelatterisdissolved,andfilterofftheexcess.Asolutioncontaining10gofpotassiumiodidein100mLdissolvesapproximately14gofHgI2at20.MercuricNitrateTS—Dissolve40gofmercuricoxide(redoryellow)inamixtureof32mLofnitricacidand15mLofwater.Storeinglasscontainers,protectedfromlight.Mercuric–PotassiumIodideTS(Mayer'sReagent)—Dissolve1.358gofmercuricchloridein60mLofwater.Dissolve5gofpotassiumiodidein10mLofwater.Mixthetwosolutions,anddilutewithwaterto100mL.Mercuric–PotassiumIodideTS,Alkaline(Nessler'sReagent)—Dissolve143gofsodiumhydroxidein700mLofwater.Dissolve50gofredmercuriciodideand40gofpotassiumiodidein200mLofwater.Pourtheiodidesolutionintothehydroxidesolution,anddilutewithwaterto1000mL.Allowtosettle,andusetheclearsupernatant.MercuricSulfateTS(Denigès'Reagent)—Mix5gofyellowmercuricoxidewith40mLofwater,andwhilestirringslowlyadd20mLofsulfuricacid,thenaddanother40mLofwater,andstiruntilcompletelydissolved.MercurousNitrateTS—Dissolve15gofmercurousnitrateinamixtureof90mLofwaterand10mLofdilutednitricacid.Storeindark,amber-coloredbottlesinwhichasmallglobuleofmercuryhasbeenplaced.MetaphenylenediamineHydrochlorideTS—Dissolve1gofmetaphenylenediaminehydrochloridein200mLofwater.Thesolutionmustbecolorlesswhenused.Ifnecessary,decolorizebyheatingwithactivatedcharcoal.Metaphosphoric–AceticAcidsTS—Dissolve15gofmetaphosphoricacidin40mLofglacialaceticacidandsufficientwatertomake500mL.Storeinacoldplace,andusewithin2days.3-Methyl-2-benzothiazolinoneHydrazoneHydrochlorideTS—Dissolve0.1gof3-methyl-2-benzothiazolinonehydrazonehydrochloridemonohydratein10mLofwater,dilutetheresultingsolutionwithmethanolto100mL,andmix.MethylOrangeTS—Dissolve100mgofmethylorangein100mLofwater,andfilterifnecessary.MethylPurpleTS—UseMethylRed–MethyleneBlueTS.MethylRedTS—Dissolve100mgofmethylredin100mLofalcohol,andfilterifnecessary.MethylRedTS,Methanolic—Dissolve1gofmethylredin100mLofmethanol,andfilter,ifnecessary.Storeprotectedfromlight,andusewithin21days.MethylRed–MethyleneBlueTS—Add10mLofmethylredTSto10mLofmethyleneblueTS,andmix.MethylVioletTS—UseCrystalVioletTS.MethylYellowTS—Prepareasolutioncontaining0.10mgpermLinalcohol.MethylYellow–MethyleneBlueTS—Dissolve1gofmethylyellowand100mgofmethylenebluein125mLofmethanol.MethyleneBlueTS—Dissolve125mgofmethylenebluein100mLofalcohol,anddilutewithalcoholto250mL.MethylthioninePerchlorateTS—To500mLofpotassiumperchloratesolution(1in1000)adddropwise,withconstantshaking,methylenebluesolution(1in100)untilaslight,permanentturbidityresults.Allowtheprecipitatetosettle,decantthesupernatantthroughpaper,anduseonlytheclearsolution.Millon'sReagent—To2mLofmercuryinaconicalflaskadd20mLofnitricacid.Shaketheflaskunderahoodtobreakupthemercuryintosmallglobules.Afterabout10minutes,add35mLofwater,and,ifaprecipitateorcrystalsappear,addsufficientdilutenitricacid(1in5,preparedfromnitricacidfromwhichtheoxideshavebeenremovedbyblowingairthroughituntilitiscolorless)todissolvetheseparatedsolid.Addsodiumhydroxidesolution(1in10)dropwise,withthoroughmixing,untilthecurdyprecipitatethatformsaftertheadditionofeachdropnolongerredissolvesbutisdispersedtoformasuspension.Add5mLmoreofthedilutenitricacid,andmix.Preparethissolutionfresh.Molybdo-phosphotungstateTS(Folin-DenisReagent)—Toabout350mLofwatercontainedinaround-bottomflaskadd50gofsodiumtungstate,12gofphosphomolybdicacid,and25mLofphosphoricacid.Boilthemixtureunderarefluxcondenserfor2hours,thencool,dilutewithwaterto500mL,andmix.Storeintightcontainers,protectedfromlight,andinacoldplace.1-NaphtholReagent—Dissolve1gof1-naphtholin25mLofmethanol.Preparethissolutionfresh.1-NaphtholTS—Use1-NaphtholReagent.2-NaphtholTS(BetanaphtholTS)—Dissolve1gof2-naphtholin100mLofsodiumhydroxidesolution(1in100).p-NaphtholbenzeinTS—Dissolve250mgofp-naphtholbenzeinin100mLofglacialaceticacid.N-(1-Naphthyl)ethylenediamineDihydrochlorideTS—Dissolve100mgofN-(1-naphthyl)ethylenediaminedihydrochloridein100mLofamixtureof7partsofacetoneand3partsofwater.Nessler'sReagent—SeeMercuric–PotassiumIodideTS,Alkaline.NeutralRedTS—Dissolve100mgofneutralredin100mLof50%alcohol.NickelStandardSolutionTS—Dissolve4.78gofnickel(II)sulfateheptahydrateinwater,anddilutewithwaterto1000mL.Immediatelypriortouse,dilute10.0mLofthesolutionsoobtainedwithwaterto1000mL.Suitablenickelstandardsolutionsarealsoavailablecommercially.NinhydrinTS—UseTriketohydrindeneHydrateTS.p-NitroanilineTS—To350mgofp-nitroanilineadd1.5mLofhydrochloricacid,andmix.Dilutewithwaterto50mL,mix,andallowtosettle.Place5mLoftheclearsupernatantina100-mLvolumetricflask,andimmerseitinanicebath.Whileitisintheicebath,add1mLofhydrochloricacid,thenadd,insmallportions,2mLofsodiumnitritesolution(1in100),dilutewithwatertovolume,andmix.NitrophenanthrolineTS—Dissolve150mgof5-nitro-1,10-phenanthrolinein15mLoffreshlypreparedferroussulfatesolution(1in140).OracetBlueBTS—A1in200solutionoforacetblueBinglacialaceticacid.OrthophenanthrolineTS—Dissolve150mgoforthophenanthrolinein10mLofasolutionofferroussulfate,preparedbydissolving700mgofclearcrystalsofferroussulfatein100mLofwater.Theferroussulfatesolutionmustbepreparedimmediatelybeforedissolvingtheorthophenanthroline.Storeinwell-closedcontainers.OxalicAcidTS—Dissolve6.3gofoxalicacidinwatertomake100mL.PalladiumChlorideTS,Buffered—Weigh500mgofpalladiumchlorideintoa250-mLbeaker,add5mLofconcentratedhydrochloricacid,andwarmthemixtureonasteambath.Add200mLofhotwaterinsmallincrementswithcontinuedheatinguntilsolutioniscomplete.Transferthesolutiontoa250-mLvolumetricflask,anddilutewithwatertovolume.Transfer50mLtoa100-mLvolumetricflask.Add10mLof1Msodiumacetateand9.6mLof1Nhydrochloricacid.Dilutewithwatertovolume.PerchloricAcidTS—Dilute8.5mLofperchloricacidwithwaterto100mL.PhenolTS—Dissolve1.2gofphenolinalcoholtomake10mL.Prepareweekly.PhenolRedTS(PhenolsulfonphthaleinTS)—Dissolve100mgofphenolsulfonphthaleinin100mLofalcohol,andfilterifnecessary.pH4.7PhenolRedTS—Dissolve33mgofphenolsulfonphthaleinin1.5mLof2Nsodiumhydroxidesolution,dilutewithwaterto100mL,andmix(SolutionA).Dissolve25mgofammoniumsulfatein235mLofwater,add105mLof2Nsodiumhydroxidesolutionand135mLof2Naceticacid,andmix(SolutionB).Add25mLofSolutionAtoSolutionB,andmix.Ifnecessary,adjustthepHofthissolutionto4.7.PhenoldisulfonicAcidTS—Dissolve2.5gofphenolin15mLofsulfuricacidinaflaskofsuitablecapacity.Add7.5mLoffumingsulfuricacid,stirwell,andheatat100for2hours.Transfertheproduct,whilestillfluid,toaglass-stopperedbottle,and,whendesiredforuse,warminawaterbathuntilliquefied.PhenolphthaleinTS—Dissolve1gofphenolphthaleinin100mLofalcohol.PhenylhydrazineAcetateTS—Dissolve10mLofphenylhydrazineand5mLofglacialaceticacidinwatertomake100mL.Phenylhydrazine–SulfuricAcidTS—Dissolve65mgofphenylhydrazinehydrochloridein100mLofacooledmixtureofequalvolumesofsulfuricacidandwater.PhloroglucinolTS—Dissolve500mgofphloroglucinolin25mLofalcohol.Storeintightcontainers,protectedfromlight.PhosphaticEnzymeTS—Dissolve5gofphosphaticenzymeinwatertomake50mL.Preparethissolutionfresh.PhosphomolybdicAcidTS—Dissolve20gofphosphomolybdicacidinalcoholtomake100mL.Filterthesolution,anduseonlytheclearfiltrate.PhosphotungsticAcidTS—Dissolve1gofphosphotungsticacidinwatertomake100mL.PicrateTS,Alkaline—Mix20mLoftrinitrophenolsolution(1in100)with10mLofsodiumhydroxidesolution(1in20),dilutewithwaterto100mL,andmix.Usewithin2days.PicricAcidTS—SeeTrinitrophenolTS.PlatinicChlorideTS—Dissolve2.6gofplatinicchlorideinwatertomake20mL.Platinum–CobaltTS—Dissolve1.246gofpotassiumchloroplatinate(K2PtCl6)and1.000gofcobaltchloride(CoCl2·6H2O)inwater,add100mLofhydrochloricacid,anddilutewithwaterto1L.PotassiumAcetateTS—Dissolve10gofpotassiumacetateinwatertomake100mL.Potassium–BismuthIodideTS—Dissolve12.5goftartaricacidin25mLofwater,thendissolve1.06gofbismuthsubnitrateinthismixture(SolutionA).Dissolve20gofpotassiumiodidein25mLofwater(SolutionB).Dissolve100goftartaricacidin450mLofwater(SolutionC).AddSolutionsAandBtoSolutionC,andmix.PotassiumCarbonateTS—Dissolve7gofanhydrouspotassiumcarbonateinwatertomake100mL.PotassiumChromateTS—Dissolve10gofpotassiumchromateinwatertomake100mL.PotassiumDichromateTS—Dissolve7.5gofpotassiumdichromateinwatertomake100mL.PotassiumFerricyanideTS—Dissolve1gofpotassiumferricyanidein10mLofwater.Preparethissolutionfresh.PotassiumFerrocyanideTS—Dissolve1gofpotassiumferrocyanidein10mLofwater.Preparethissolutionfresh.PotassiumHydroxideTS—Dissolve6.5gofpotassiumhydroxideinwatertomake100mL.PotassiumHydroxideTS,Alcoholic—Use0.5NPotassiumHydroxide,Alcoholic(seeinthesectionVolumetricSolutions).PotassiumIodideTS—Dissolve16.5gofpotassiumiodideinwatertomake100mL.Storeinlight-resistantcontainers.PotassiumIodideandStarchTS—Dissolve0.75gofpotassiumiodidein100mLofwater.Heattoboiling,andadd,withstirring,asolutionof0.5gofsolublestarchin35mLofwater.Boilfor2minutes,andallowtocool.Sensitivity—Mix15mLin0.05mLofglacialaceticacidand0.3mLofdilutediodineTS:abluecolorisproduced.PotassiumIodoplatinateTS—Dissolve200mgofplatinicchloridein2mLofwater,mixwith25mLofpotassiumiodidesolution(1in25),andaddwatertomake50mL.PotassiumPermanganateTS—Use0.1NPotassiumPermanganate(seeinthesectionVolumetricSolutions).PotassiumPyroantimonateTS—Dissolve2gofpotassiumpyroantimonatein85mLofhotwater.Coolquickly,andadd10mLofasolutionofpotassiumhydroxide(3in20).Allowtostandfor24hours,filter,anddilutewithwaterto100mL.PotassiumSulfateTS—Dissolve1gofpotassiumsulfateinwatertomake100mL.PotassiumThiocyanateTS—Dissolve9.7gofpotassiumthiocyanateinwatertomake100mL.Pyridine–PyrazoloneTS—To100mLofasaturatedsolutionof1-phenyl-3-methyl-2-pyrazoline-5-oneadd20mLofa1in1000solutionof3,3-dimethyl-1,1-diphenyl-[4,4-bi-2-pyrazoline]-5,5-dioneinpyridine.Storeinadarkbottle,andusewithin3days.PyrogallolTS,Alkaline—Dissolve500mgofpyrogallolin2mLofwater.Dissolve12gofpotassiumhydroxidein8mLofwater.Thesolutionsshouldbefreshlypreparedandmixedimmediatelybeforeuse.QuinaldineRedTS—Dissolve100mgofquinaldineredin100mLofalcohol.QuinoneTS—Dissolve500mgofp-benzoquinonein2.5mLofglacialaceticacid,anddilutewithalcoholto50mL.Preparethissolutionfreshdaily.ResorcinolTS—Dissolve1gofresorcinolinhydrochloricacidtomake100mL.RutheniumRedTS—Dissolve10gofleadacetateinwater,dilutewithwaterto100mL,andadd80mgofrutheniumred.Thesolutioniswine-redincolor.[note—Ifnecessary,addadditionalrutheniumredtoobtainawine-redcolor.]SalineTS—Dissolve9.0gofsodiumchlorideinwatertomake1000mL.note—Wherepyrogen-freesalineTSisspecifiedinthisPharmacopeia,salineTSthathasmettherequirementsofthePyrogenTest151istobeused.SalineTS,Pyrogen-Free—SeeSalineTS.Schweitzer'sReagent—SeeCupricOxide,Ammoniated,TS.Silver–Ammonia–NitrateTS—Dissolve1gofsilvernitratein20mLofwater.AddammoniaTS,dropwise,withconstantstirring,untiltheprecipitateisalmostbutnotentirelydissolved.Filter,andstoreintight,light-resistantcontainers.Silver–AmmoniumNitrateTS—SeeSilver–Ammonia–NitrateTS.SilverDiethyldithiocarbamateTS—Dissolve1gofsilverdiethyldithiocarbamatein200mLofpyridinefromafreshlyopenedbottleorthatwhichhasbeenrecentlydistilled.Storeinlight-resistantcontainers,andusewithin30days.SilverNitrateTS—Use0.1NSilverNitrate(seeinthesectionVolumetricSolutions).SimulatedGastricFluidTS—SeeGastricFluid,Simulated,TS.SimulatedIntestinalFluidTS—SeeIntestinalFluid,Simulated,TS.SodiumAcetateTS—Dissolve13.6gofsodiumacetateinwatertomake100mL.SodiumAlizarinsulfonateTS—Dissolve100mgofsodiumalizarinsulfonatein100mLofwater,andfilter.SodiumAminoacetateTS(SodiumGlycinateTS)—Dissolve3.75gofaminoaceticacidinabout500mLofwater,add2.1gofsodiumhydroxide,anddilutewithwaterto1000mL.Mix9mLoftheresultingsolutionwith1mLofdiluteglacialaceticacid(1in300).ThistestsolutionhasapHbetween10.4and10.5.SodiumBisulfiteTS—Dissolve10gofsodiumbisulfiteinwatertomake30mL.Preparethissolutionfresh.SodiumBitartrateTS—Dissolve1gofsodiumbitartrateinwatertomake10mL.Preparethissolutionfresh.SodiumCarbonateTS—Dissolve10.6gofanhydroussodiumcarbonateinwatertomake100mL.SodiumChlorideTS,Alkaline—Dissolve2gofsodiumhydroxidein100mLofwater,saturatethesolutionwithsodiumchloride,andfilter.SodiumCitrateTS—Dissolve73.5gofsodiumcitratedihydrateinwatertomake250mL.SodiumCitrateTS,Alkaline—Dissolve50gofsodiumcitratedihydrateand2.5gofsodiumhydroxideinwatertomake250mL.SodiumCobaltinitriteTS—Dissolve10gofsodiumcobaltinitriteinwatertomake50mL,andfilterifnecessary.SodiumFluorideTS—Dryabout500mgofsodiumfluorideat200for4hours.Accuratelyweigh222mgofthedriedmaterial,anddissolveinwatertomake100.0mL.Pipet10mLofthissolutionintoa1-Lvolumetricflask,anddilutewithwatertovolume.EachmLofthissolutioncorrespondsto0.01mgoffluorine(F).SodiumHydrosulfiteTS,Alkaline—Dissolve25gofpotassiumhydroxidein35mLofwater,and50gofsodiumhydrosulfitein250mLofwater.Whenthetestsolutionisrequired,mix40mLofthehydroxidesolutionwiththe250mLofthehydrosulfitesolution.Preparethissolutionfresh.SodiumHydroxideTS—Dissolve4.0gofsodiumhydroxideinwatertomake100mL.SodiumHydroxideTS2—Transfer8.5gofsodiumhydroxidetoa100-mLvolumetricflask,anddissolveinanddilutewithwatertovolume.SodiumHypobromiteTS—Toasolutionof20gofsodiumhydroxidein75mLofwateradd5mLofbromine.Aftersolutionhastakenplace,dilutewithwaterto100mL.Preparethissolutionfresh.SodiumHypochloriteTS—UseSodiumHypochloriteSolution(seeinthesectionReagentSpecifications).SodiumIodohydroxyquinolinesulfonateTS—Dissolve8.8gofiodohydroxyquinolinesulfonicacidin200mLofwater,andadd6.5mLof4Nsodiumhydroxide.Dilutewithwaterto250mL,mix,andfilter.SodiumNitroferricyanideTS—Dissolve1gofsodiumnitroferricyanideinwatertomake20mL.Preparethissolutionfresh.DibasicSodiumPhosphateTS—Dissolve12gofclearcrystalsofdibasicsodiumphosphateinwatertomake100mL.SodiumPhosphotungstateTS—Toasolutionof20gofsodiumtungstatein100mLofwateraddsufficientphosphoricacidtoimpartastronglyacidreactiontolitmus,andfilter.Whenrequiredforuse,decanttheclearsolutionfromanysedimentthatmaybepresent.Storeintight,light-resistantcontainers.SodiumSulfideTS—Dissolve1gofsodiumsulfideinwatertomake10mL.Preparethissolutionfresh.SodiumTartrateTS—Dissolve11.5gofsodiumtartrateinwatertomake100mL.SodiumTetraphenylboronTS—Dissolve1.2gofsodiumtetraphenylboroninwatertomake200mL.Ifnecessary,stirfor5minuteswith1gofaluminumoxide,andfiltertoclarify.SodiumThioglycolateTS—Dissolve1.5gofsodiumthioglycolatein450mLofwater,andadd50mLofalcohol.Usewithin3days.SodiumThiosulfateTS—Use0.1NSodiumThiosulfate(seeinthesectionVolumetricSolutions).StandardLeadSolution—SeeunderHeavyMetals231.StannousChloride,Acid,TS—Dissolve8gofstannouschloridein500mLofhydrochloricacid.Storeinglasscontainers,andusewithin3months.StannousChloride,Acid,Stronger,TS—Dissolve40gofstannouschloridein100mLofhydrochloricacid.Storeinglasscontainers,andusewithin3months.Iodide-FreeStarchTS—Mix1gofsolublestarchwithsufficientcoldwatertomakeathinpaste.Whilestirring,add100mLofboilingwater,andallowtocool.Preparethissolutionimmediatelybeforeuse.Iodide-freestarchTSshowsabluecolorwhen20mLofpotassiumiodidesolution(1in400)and0.05mLofaniodine–potassiumiodidesolution(preparedbydissolving127mgofiodineand800mgofpotassiumiodideinwateranddilutingwithwaterto100mL)areaddedto1mLoftheiodide-freestarchTS.StarchIodidePasteTS—Heat100mLofwaterina250-mLbeakertoboiling,addasolutionof0.75gofpotassiumiodidein5mLofwater,thenadd2gofzincchloridedissolvedin10mLofwater,and,whilethesolutionisboiling,add,withstirring,asmoothsuspensionof5gofsolublestarchin30mLofcoldwater.Continuetoboilfor2minutes,thencool.Storeinwell-closedcontainersinacoldplace.StarchiodidepasteTSmustshowadefinitebluestreakwhenaglassrod,dippedinamixtureof1mLof0.1Msodiumnitrite,500mLofwater,and10mLofhydrochloricacid,isstreakedonasmearofthepaste.Starch–PotassiumIodideTS—Dissolve500mgofpotassiumiodidein100mLoffreshlypreparedstarchTS.Preparethissolutionfresh.StarchTS—Mix1gofsolublestarchwith10mgofredmercuriciodideandsufficientcoldwatertomakeathinpaste.Add200mLofboilingwater,andboilfor1minutewithcontinuousstirring.Cool,anduseonlytheclearsolution.[note—Commerciallyavailable,stabilizedstarchindicatorsolutionsmaybeused.]StrongerCupricAcetateTS—SeeCupricAcetateTS,Stronger.SudanIIITS—Dissolve0.05gofSudanIIIin25mLofalcohol,withwarmingifnecessary.Cool,add25mLofglycerin,andmix.Filterifundissolvedmaterialpersists.SudanIVTS—Dissolve0.5gofSudanIVinchloroformtomake100mL.SulfanilicAcidTS—Dissolve800mgofsulfanilicacidin100mLofaceticacid.Storeintightcontainers.DiazotizedSulfanilicAcidTS—Dissolve0.9gofsulfanilicacidin9mLofhydrochloricacidwithwarming,anddilutewithwaterto100mL.Cool10mLofthissolutioninicedwater,andadd10mLofasodiumnitritesolution(4.5in100)previouslycooledinicedwater.Allowtostandat0foratleast15minutes(thesolutionmaybekeptfor3daysatthistemperature).Immediatelybeforeuse,add20mLofsodiumcarbonatesolution(1in10).Sulfanilic--NaphthylamineTS—SeeSulfanilic-1-NaphthylamineTS.Sulfanilic-1-NaphthylamineTS—Dissolve500mgofsulfanilicacidin150mLofaceticacid.Dissolve100mgof1-naphthylaminehydrochloridein150mLofaceticacid,andmixthetwosolutions.Thepinkcolorthatmaydeveloponstandingcanberemovedbytreatmentwithzinc.SulfomolybdicAcidTS—Dissolve,withtheaidofheat,2.5gofammoniummolybdatein20mLofwater,add50mLof12Nsulfuricacid,anddilutewithwaterto100mL.Storethissolutioninapolyethylenecontainer.SulfuricAcidTS—Addaquantityofsulfuricacidofknownconcentrationtosufficientwatertoadjustthefinalconcentrationtobetween94.5%and95.5%(w/w)ofH2SO4.note—Sincetheacidconcentrationmaychangeuponstandingoruponintermittentuse,theconcentrationshouldbecheckedfrequentlyandsolutionsassayingmorethan95.5%orlessthan94.5%discarded.SulfuricAcid–FormaldehydeTS—Add1dropofformaldehydeTStoeachmLofsulfuricacid,andmix.Preparethissolutionfresh.TannicAcidTS—Dissolve1goftannicacidin1mLofalcohol,anddilutewithwaterto10mL.Preparethissolutionfresh.TartaricAcidTS—Dissolve3goftartaricacidinwatertomake10mL.Preparethissolutionfresh.TetrabromophenolphthaleinEthylEsterTS—Dissolve100mgoftetrabromophenolphthaleinethylesterin90mLofglacialaceticacid,anddilutewithglacialaceticacidto100mL.Preparethissolutionfresh.TetramethylammoniumHydroxideTS—Useanaqueoussolutioncontaining,ineach100mL,theequivalentof10gofanhydroustetramethylammoniumhydroxide.ThioacetamideTS—Dissolve4gofthioacetamidein100mLofwater.Thioacetamide–GlycerinBaseTS—Mix0.2mLofthioacetamideTSand1mLofglycerinbaseTS,andheatinaboilingwaterbathfor20seconds.Usethemixtureimmediately.ThoriumNitrateTS—Dissolve1gofthoriumnitrateinwatertomake100mL.Filter,ifnecessary.ThymolBlueTS—Dissolve100mgofthymolbluein100mLofalcohol,andfilterifnecessary.ThymolphthaleinTS—Dissolve100mgofthymolphthaleinin100mLofalcohol,andfilterifnecessary.TitaniumTrichlorideTS—Dissolve15goftitaniumtrichloridein100mLof10%hydrochloricacidsolution.TitaniumTrichloride–SulfuricAcidTS—Mixcarefully20mLoftitaniumtrichlorideTSin13mLofsulfuricacid.Addsufficient30%hydrogenperoxidetoproduceayellowcolor.Heatuntilwhitefumesareevolved,allowtocool,anddilutewithwater.Repeattheevaporationandadditionofwateruntilacolorlesssolutionisobtained.Dilutewithwaterto100mL.p-ToluenesulfonicAcidTS—Dissolve2gofp-toluenesulfonicacidin10mLofamixtureof7partsofacetoneand3partsofwater.TriketohydrindeneHydrateTS(NinhydrinTS)—Dissolve200mgoftriketohydrindenehydrateinwatertomake10mL.Preparethissolutionfresh.TrinitrophenolTS(PicricAcidTS)—Dissolvetheequivalentof1gofanhydroustrinitrophenolin100mLofhotwater.Coolthesolution,andfilterifnecessary.TriphenyltetrazoliumChlorideTS—Dissolve500mgoftriphenyltetrazoliumchlorideindehydratedalcoholtomake100mL.XylenolOrangeTS—Dissolve100mgofxylenolorangein100mLofalcohol.ZincUranylAcetateTS—Dissolve50gofuranylacetateinamixtureof15mLofglacialaceticacidandwatertomake500mL.Thendissolve150gofzincacetateinamixtureof15mLofglacialaceticacidandwatertomake500mL.Mixthetwosolutions,allowtostandovernight,andpassthroughadryfilter,ifnecessary.