AmiconStirredUltrafiltrationCells(Models8050,8400)Description-Membranes-Assembly-Operation-Limits-Sterilization-Maintenance-Troubleshooting
Description
Forproteinconcentration,gaspressureisapplieddirectlytoultrafiltrationcell.Solutesabovethemembrane"smolecularweight(MW)cut-offareretainedincell,whilewaterandsolutesbelowthecut-offpassintothefiltrateandoutofcell. | 8050 | 8400 |
| Cellcapacity(ml)Stirredminimumvolume¹(ml)Membranediameter(mm)Effectivemembranearea(cm²)Hold-upvolume²(ml) | 502.54313.40.5 | 40010.07641.81.5 |
| Maximumoperatingpressure:75psi(5.3kg/cm²)Reliefvalvesetting:90psi(6.3kg/cm²)Maximumdiafiltrationoperatingpressure:55psi(3.9kg/cm²) |
| ¹Minimumworkingvolume.²Non-recoverablevolume(belowmembranesurface). |
Membranes
- YM10Ø43mm(for8050),Amicon#13622,10pack:$108YM10Ø76mm(for8400),Amicon#13642,10pack:$139
- Prewash:floatmembranewithglossysidedowninbeakerwithdistilledwaterforatleast1hourchangingwater3times.Allmembranesarepretreatedwithglycerinetopreventdrying,YMmembranesaretreatedwithsodiumazideasapreservative.
- Cleaning:rinsewith0.1MNaOH,followedbythoroughflushingwithdistilledwater.Forstronglyabsorbedprotein,soakin0.1%proteasesolutionandrinsethoroughly.
- Storage:forreuse,storemembranesin10%ethanolwatersolutionat4°C.
Assembly
- Placemembraneinholder,shinysideup;thenplaceO-ringontopofmembrane.GentlypushO-ringdownsothatitcontactsandseatsmembraneevenlyinbottomofholder.Handlemembranebyitsedgestoavoidscratchingorcontaminatingsurface.
- Fitmembraneholderintocellbody,aligningtabsonsidesofholderwithslotsinbaseofcellbody.
- Invertcellbodyandmembraneholder;screwbasefirmlyintobottomofcellbody.Adefinite"stop"willbefeltwhenbaseandbodyarefullyengaged.Topofmembraneholderwillbeflushwithbottomofslotsincellbody.
- Pushfiltrateexittubingontoexitspoutofmembranesupport.
- Placestirrerassemblyintocellbody.Whenproperlyinstalled,armsofstirrerassemblywillrestonsmallridgeinsidetopofcellbody.
- Introducesampleintocell.
- Withatwistingmotion,pushcapdownontocellbody,orientinggasinletportoncapoppositefiltrateexitportonholder.Ifcapassemblydoesnotslideeasily,lubricateO-ringlightlywithpetroleumjelly.Donotallowpetroleumjellytocontactmembrane!
- Setpressure-reliefvalvetohorizontal(open)position.
- Slidecellintoretainingstand,fittingringoncellbaseintoholeinstand.Flattenededgesonbottomflangeofcapensuresthatcellisinsertedproperlyandpreventsrotationofcellonceinsidestand.
- Turnpressure-reliefvalvetovertical(closed)position.
- Attachgaspressureline.
Operation
- Placecellonmagneticstirringtable.
- Connectinletlinetoregulatedgaspressuresource(Nitrogengasisrecommendedforpressurizingcell!UseofcompressedaircancauselargepHshiftsduetodissolutionofcarbondioxide.Withsensitivesolutions,oxidationcanalsooccur,leADIngtootherpotentialproblems.)
- Holdcellsteadyonthestirringtableandpressurizeaccordingtoinstructionsinmembranepackage.Generally55psi(3.7kg/cm2)isoptimal,maximal70psi(4.7kg/cm2)nitrogengaspressure.Capassemblymovesupward,formingasecurelockwithretainingstandoncesystemispressurized.
- Turnonstirringtableandadjuststirringrateuntilthevortexcreatedisapproximatelyone-thirdthedepthoftheliquidvolume.
- Monitorconcentration.Donotallowretentatetorunthrough.
- Collectpermeate.
- Highlyviscoussolutionsfilterslowly,asdosolutionscontainingparticulatematter,suchascolloids.Wheretheviscousagent(sucrose,glycerin,etc.)istoberemoved,flowcanoftenbeincreasedbypredilution.
- Prefilterorcentrifugeanysolutioncontainingparticulatematter,suchascelldebrisorprecipitates.
- Tomaximizerecoveryofretainedsubstances,continuestirringforafewminutesafterdepressurization.ThiswillresUSPendthepolarizedlayeratthemembranesurface.
- Whenfinished,turnoffnitrogenpressuresourceandstirringtable.
- Ventpressureinsidecellbyslowlyturningpressure-reliefknobtohorizontalposition.Pushcapdown,thenslidecelloutfromretainingstand.Note:Overlyrapiddepressurizationcancausethemembranetobuckleupandrupture.
- Usingatwistingmotion,removecellcapandthemagneticstirrerassembly.Alwaysremovethecelltopwiththepressure-reliefvalvesettothehorizontal(open)position.Removalinvertical(closed)positioncancreateapartialvacuumwhichcanrupturethemembrane!Pouroutsolution.
- Disassemblecell,thenwashallcomponentswithamilddetergent/watersolution,thenrinsethoroughly.Causticcleaningsolutionsmaydamagetheanodizedaluminumretainingstand.Leaveultrafiltrationcelldisassembledwheneveritisunlikelytobeusedforseveralweeks.
Limits
- max.celloperatingpressure:75psi(5.3kg/cm2)
- max.membraneoperatingpressure:70psi(4.7kg/cm2)
- pressure-reliefvalvepreset:90psi(6.3kg/cm2)
- AlthoughbriefexposuretohighertemperaturesispossIBLe,donotoperatecellcontinuouslyabove85°C(185°F).
- pH2to10
- Donotusethefollowingchemicals:Ketones(includingacetone),Aromatichydrocarbons(includingtoluene),Cellosolvers,Halogenatedhydrocarbons,DMF,Aliphaticesters,DMSO,PolarAromatics.Note:Thespringinthepressure-reliefvalveisnotcompatiblewith0.1NNaOH.
Sterilization
Amiconstirredultrafiltrationcellscanbeautoclavedat121°C(250°F)for30minutes.Theyarecompatiblewithstandardsterilizinggasmixtures,70%ethanol,isopropanolor5%formalin.CAUTION:Tightenthebasepartiallybeforeautoclaving.
Maintenance
- ReplaceO-ringsatthefirstsignofdamageorwear.
- Ifinstallationorremovalofcapassemblybecomesdifficult,lubricatecapO-ringwithasmallamountofpetroleumjelly.Donotallowpetroleumjellytocomeincontactwithmembrane!
- Checkstirrerperiodicallyforroughedgesthatcouldpossiblydamagethemembrane.
- Replacetransparentbodyimmediatelyshoulditbecomecrackedorcrazed.
Troubleshooting
Littleornofiltrateobtained:- Rotatepressure-reliefvalvetocheckforpressure.Ifnotpressurized,checknitrogensourceandregulator.
- Makesureglossysideofdiscmembranefacesup.
- Ifsamplesolutionishighlyviscousduetomicrosolutes,eitherdiluteordiafiltertoincreaseflowrate.
- Checkmembraneholderandfiltrateportforblockage.
Filtraterateabnormallyhigh:- Checkmembraneforlesions,scratches,orroughness.
- Makesurecorrectmembranetypeisbeingused.
- Checkstirrerassemblytoensurethatstirringbarisnotcontactingthemembranesurface.
Cellleaks:- BesurelowerO-ringrestsentirelyonthemembraneperipheralsurface.
- CheckO-ringsfornicksandcuts.
- Makesuremembranesupportisseatedproperly,andthatthebaseisscrewedinfirmly.
- MakesureO-ringsarenotsqueezedoutofslots.
- Checkfittingsongasinlettubingforcorrectorderandposition.
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