Sample Preparation Solutions - Fisher Sci
1.51.00.5Thermo ScientificTechnical Resources DocumentSample preparationsolutions
Sample PreparationSection ContentsSPE Cartridge Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-002SPE Phase Selection by Manufacturer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-003Removing Uncertainty by Applying the Science to SPE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-004SPE Procedure - Six Steps for a Clean Extract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-005Method Optimization in SPE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-006Syringes and Filtration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-007Thermo Scientific Chromatography Columns and Consumables 2016-2017Syringe Filter Membrane Compatibility Chart . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-0081-001
SPE Cartridge SelectionThermo Scientific SPE products have been developed to meet the requirements of today’s sample preparationchallenges. Thermo Scientific SPE cartridges and 96-well plates are offered in a range of phases and bed weights,ideal for use in application areas such as: Pharmaceutical, Biochemical, Environmental, and Food and Beverage.The following flow chart is designed to help in selecting the correct phase for the compound of interest.SPE Phase SelectionSampleMatrixSolubilityof SamplePolarityof SampleSeparationModeSample Preparation Thermo Scientific SPE ProductsRecommendedHyperSep Sorbent(s)SOLA HRP, C18, C8,Phenyl, Retain PEPSOLA HRP, C18, C8,Retain PEPSOLA HRP, HypercarbRetain PEPSCXSAXAminopropylSOLA SCX/WCX, Verify CXRetain CXSOLA SAX/WAX, Verify AXRetain AXSOLA HRP, C18, C8,Phenyl, Retain PEPSilica, AminopropylCyano, DiolSilica, Florisil, AminopropylCyano, DiolSCXSAXAminopropylVerify CXVerify AXVisit our online product selector tool .com/chromatography1-002
Sample PreparationSPE Phase Selection by ManufacturerThermo Scientific HyperSep ProductHyperSep Retain PEPOasis HLB (Waters)Strata-X (Phenomenex)Bond Elut Plexa (Agilent)Easy (Macherey Nagel)ISOLUTE ENV (Biotage)SampliQ OPTStyreScreen DVB (UCT)H2 O-philic DVB (JT Baker)HyperSep Retain CXOasis MCX (Waters)Strata-X-C (Phenomenex)Bond Elut Plexa PCX (Agilent)StyreScreen DBX (UCT)H2 O-philic SC-DVB (JT Baker)HyperSep Retain AXOasis MAX (Waters)Bond Elut Plexa PAX (Agilent)StyreScreen QAX (UCT)H2 O-philic SA-DVB (JT Baker)HyperSep HypercarbSupelClean ENVI-Carb Bond Elut CarbonHyperSep C18CLEAN-UP C18-USupelclean ENVI-18 / LC-18BAKERBOND SPE Polar Plus ISOLUTE C18CHROMABOND C18Bond Elut C18Strata C18-USep-Pak C18HyperSep C8Thermo Scientific Chromatography Columns and Consumables 2016-2017HyperSep Phenyl1-003Alternative ToHyperSep SilicaCLEAN-UP C8Supelclean ENVI-8 / LC-8BAKERBOND SPE Octyl C8ISOLUTE C8CHROMABOND C8Bond Elut C8Strata C8Sep-Pak C8CLEAN-UP PhenylSupelclean LC-PhBAKERBOND SPE PhenylISOLUTE PhBond Elut PhStrata Phenyl (PH)CLEAN-UP SilicaSupelclean LC-SiBAKERBOND SPE Silica GelISOLUTE SilicaCHROMABOND SiOHBond Elut SiStrata Si-1Sep-Pak SiThermo ScientificHyperSep ProductAlternative ToHyperSep SCXCLEAN-UP BCXSupelclean LC-SCXBAKERBOND SPE Aromatic Sulfonic AcidISOLUTE SCXCHROMABOND SABond Elut SCXStrata SCXHyperSep SAXCLEAN-UP QAXSupelclean LC-SAXBAKERBOND SPE Quaternary amineISOLUTE SAXCHROMABOND SBBond Elut SAXStrata SAXHyperSep Verify CXCLEAN-UP DAUDiscovery DSC-MCAXISOLUTE HCXCHROMABOND DrugBond Elut Certify IStrata Screen-CHyperSep Verify AXCLEAN-UP THCISOLUTE HAXBond Elut Certify IIStrata Screen-AHyperSep FlorisilCLEAN-UP FlorisilSupelclean ENVI-Florisil / LC-FlorisilBAKERBOND SPE FlorisilISOLUTE FlorisilCHROMABOND FlorisilBond Elut FlorisilStrata FL-PRSep-Pak FlorisilHyperSep AminopropylCLEAN-UP AminopropylSupelclean LC-NH 2BAKERBOND SPE AminoISOLUTE NH2CHROMABOND NH 2Bond Elut NH2Strata NH2Sep-Pak NH2HyperSep CyanoSupelclean LC-CNBAKERBOND SPE CyanoCHROMABOND CNBond Elut CyanoSep-Pak CyanopropylStrata CNHyperSep DiolSupelclean LC-DiolBAKERBOND SPE DiolCHROMABOND OHBond Elut DiolSep-Pak Diol
Our comprehensive range of SPE solutions offer unparalleled performancein purity of extract and reproducibility. Having a fundamental effect on the quality,time and analysis cost, SPE is a critical step during the sample analysis procedure.We are dedicated to supplying the highestquality SPE solutions, in combination withproviding our customers with the supportand resources to optimize their SPE solutionsand maximize their analysis.The Importance of SPEReducing the effects of the matrix on theseparation (GC/LC) and detection (UV, MS etc.)is beneficial. The use of SPE as a samplepreparation technique can significantly improveanalysis aiding robustness and generatingreproducible accurate, precise and sensitiveanalytical methods. This relies on the abilityof SPE to reproducibly:Maximize detection selectivity Reduce ion suppression Reduce protein binding Reduce matrix interferences by eliminationof matrix and particulates Compatibility of solvent with analyticaltechniqueMatrix effects are an issue in many areas ofanalytical chemistry, however, modification ofionization (ion suppression/enhancement) canbe a major problem in atmospheric pressureionization (API) mass spectrometry and, inparticular, electrospray based ion sources.Sample Preparation Removing Uncertainty by Applying the Science to SPEImprove analytical system performance Longer column lifetimes Less detector maintenance Autosampler syringes less likely to block Less contaminationImprove sensitivity by concentrationof the analytes Lower limits of detection More accurate quantitation Improved data -004
Sample PreparationSPE Procedure – Six Steps for a Clean Extract1. Sample Pre-treatment2. Column ConditioningRemove impurities bound less strongly thanthe compounds of interest. Use the same solvent as for samplepre-treatmentPrevent sorbent drying during columnequilibration4. Sample Loading4. Sample Loading5. Wash Away Interferences6. Elute Compounds of Interest1. Sample Pre-treatmentIt is important to optimize the samplefor effective analyte retention. The followingshould be considered: Thermo Scientific Chromatography Columns and Consumables 2016-20175. Wash Away InterferencesEquilibrate with weakly eluting solvent toprepare the phase for sample addition. 3. Column Pre-equilibration1-0053. Column Pre-equilibration Sample volume/analyte concentration/matrix complexityAdjust sample/matrix composition for properdilution/ionic strengthSample pH for optimum retentionConfirm that analytes are free in solutionRemove any unwanted particulates viafiltration or centrifugation2. Column ConditioningPrepare the sorbent for effective interaction(s)with the compounds of interest. Use appropriate solvent for columncondition/activationPrevent sorbent drying during conditioningAnalytes are retained on the sorbent. Apply samples at appropriate flow rate(1mL/minute typical)For Reversed-Phase Interactions Neutral compounds are not affected by pH For charged compounds, a pH at which thecompound is not charged is used. Neutralizethe molecule according to the following:– For basic compounds, the neutralmolecule exists at least 2pH units belowthe pKa of the compound– For acidic compounds, the neutralmolecule exists at least 2pH units abovethe pKa of the compoundFor Normal-Phase Interactions pH is not normally an issue in normal phaseinteractions, as the solvents used aretypically non-polar organic solvents, ratherthan water There is no need to verify the sampleapplication pHFor Ion-Exchange Interactions pH and pKa are important considerations Acidic compounds are extracted from asample solution at least 2pH units abovethe pKa of the analyte Basic compounds are extracted from asample solution 2 or more pH units belowthe pKa of the analyte For second (organic) wash, choose thestrongest solution where no compoundbreakthrough occurs For elution step, use a solution stronger thanwhere all the compound of interest is eluted NB: when choosing these solutions allowsome margin for error Select a strong enough wash solventto remove interferences but weak enoughto leave compounds of interest boundSelectively rinse away the less stronglybonded interferencesWash solvent selected according to phasemechanism/analyte properties6. Elute Compounds of InterestSelectively recover the analyte(s) by disruptingthe analyte-sorbent interaction. Selectively elute analytes of interest usingdifferent solventsSmaller elution volume produces a moreconcentrated extractSelect elution solvent that does not elutestrongly retained impuritiesSelect elution solvent according to phasemechanism/analyte propertiesIt is important to optimize the Wash andElution steps in order to obtain maximumlevels of recovery.Get your copy of our applicationnotebook, featuring over150 SPE applicationswww.thermoscientific.com/spe
Sample Preparation Method Development Optimization in SPEFor cleaner extracts, optimization of the SPE process can be important.By optimizing the load, wash, and elution steps of the SPE process,a cleaner sample extract can be obtained.An example of this is in the development of an SPE method for testosterone.HyperSep Retain PEP SPE Cartridge(Part Number 60107-201)1. Condition with 1mL methanol followed by 1mL water2. Load 1mL of 500ng/mL sample in water3. Sequentially wash with increasing strengths of methanol in water,collecting the eluent 0% methanol/100% water to 90% methanol/10% water,increasing methanol content by 10% each time4. Elution with multiple volumes of methanolLogP 3.6 Pka 2.99Elution Profile of Testosterone3 x 100%2 x 100%1 x 009080706050403020100By following this procedure the optimum wash and elution steps can be determined. In subsequent analyses a single washstage would be used at 50% methanol followed by a single elution step at 90% methanol. This ensures simplicity of theprocess, optimum recovery and cleanliness of extract.An alternative would be to use a HyperSep Retain AX SPE cartridge with the added benefit of Phospholipid removal.For more information on how to optimize you SPE methodologyand specific applications visit www.thermoscientific.com/speand request your free copy of the Thermo Scientific HyperSepColumns Application 006
Sample PreparationThermo Scientific Syringes and FiltersTitan3 and Target2 Syringe FiltersSample preparation with Thermo Scientific Titan3 and Thermo Scientific Target2 syringe filters helpprovide consistent and reliable experimental results. Both these products provide high quality filtrationsolutions for a range of samples and applications. The premium Titan3 range provides even higher levelsof confidence due to the robust design characteristics (burst pressures of 120psi for the 30mm range)cleaner extracts due to the inclusion of a pre-filter (most 30mm products) and ease of membraneselection via the color coded ring.Membrane Selection GuideChoose a filter or membrane based on:1.2.3.4.Chemical compatibility of the membrane and housing with your sample matrixSize and amount of particulates in the samplePotential interactions (binding) between the membrane and sample componentsSpecial considerations such as requirement for pre-filter or inorganic ion certificationHousings Titan3 and Target2 filter housings are manufactured from solvent-resistant, low-extractablepolypropylene resins specifically selected for wide compatibility with common HPLC sample matricesSolutions at temperatures up to 100 C can be filtered using Target2 and Titan3 syringe filters.Syringe filters can be sterilized by autoclave at 125 C for 15 minutesThe inlet connection is an enhanced female Luer Lock fitting designed for extra security whenattached to a Luer Lock syringeThe outlet fitting is a standard size male Luer-slip fitting for ease of filtrate collectionTarget2 polypropylene syringe filter housings meet the requirements of 21 CFR 177.1520This table offers general guidelines for membrane characteristics and compatible applications.Membrane TypeCellulose AcetateThermo Scientific Chromatography Columns and Consumables 2016-2017Glass MicroFiberNylonPolyethersulfone (PES)PolypropylenePTFEPVDFRegenerated Cellulose1-007Membrane CharacteristicsLow protein binding, ideal for aqueous-based samples;high protein recovery from filtrate; lower protein bindingcompared to PVDFLarger porosity; able to remove large particulateswithout cloggingMost frequently selected membrane; broad compatibilitywith aqueous and organic; naturally hydrophilic membrane;extremely low in extractables; excellent flowrate with mostsample matrices; not compatible with strong acids or basesHigh flowrates with good throughput volume; low proteinbinding; compatible with high temperature liquids;mechanically strong membrane low in inorganicextractable ionsHydrophobic membrane has wide chemical compatibilitywith organic solvents; low nonspecific protein bindingHydrophobic membrane is resistant to nearly all solvents,acids, and bases; membrane is mechanically strong andwill withstand exposure to high temperature liquids; low inextractables; PTFE blocks water vapor; can be used to filteraqueous solutions after prewetting with an alcohol. Thehydrophilic PTFE option provides the same application andperformance characteristic, but does not require prewettingof the membrane when filtering aqueous samplesHydrophilic membrane with good solvent resistance; lowUV absorbing extractables and low nonspecific bindingHydrophilic membrane with good solvent resistance,extremely low nonspecific binding; compatible with nearlyall common HPLC solvents; tolerates aqueous samplesin pH range of 3 to 12ApplicationsTissue culture media filtration, sensitive biological samplesDissolution testing, general filtrationGeneral laboratory filtration; filtration for most samples;HPLC samplesNOTE: Nylon binds protein, do not use when high proteinrecovery is desiredPES is certified for ion chromatography; tissue culturefiltration; filtration of proteins and nucleic acidsFiltration of biological samples; filtration of aggressiveorganic solutionsFiltration of aggressive organic, highly basic or hot solutions,ideal for transducer protectorsGeneral biological filtration; filtration of samples wherehigh protein recovery is desiredMembrane of choice for low nonspecific binding applications;Tissue Culture media filtration and general biologicalsample filtration
Sample Preparation Syringe Filter Membrane Compatibility ChartUse the information in this table to determine the ability of a specific syringe filtermembrane to withstand exposure to a solvent. All concentrations are 100% unless noted.LegendC CompatibleLC Limited Compatibility (Membrane may swell and shrink)IC Incompatible (Not Recommended)ND No Compatibility Data Currently AvailablePTFE PolytetrafluoroethylenePVDF Polyvinylidene FluoridePES PolyethersulfoneCA Cellulose AcetateRC Regenerated CellulosePP PolypropyleneGMF Glass MicroFiberChemicalAcidsAcetic, GlacialAcetic, 25%Hydrochloric, ConcentratedHydrochloric, 25%Sulfuric, ConcentratedSulfuric, 25%Nitric, ConcentratedNitric, 25%Phosphoric, 25%Formic, 25%Trichloroacetic, 10%AlcoholsMethanol, 98%Ethanol, 98%Ethanol, 70%Isopropanoln-PropanolAmyl Alcohol (Butanol)Benzyl AlcoholEthylene GlycolPropylene GlycolGlycerolAlkaliesAmmonium Hydroxide, 25%Sodium Hydroxide, 3NAmines and y1-008
Sample PreparationThermo Scientific Chromatography Columns and Consumables 2016-20171-009ChemicalEstersEthyl Acetate/Methyl AcetateAmyl Acetate/Butyl AcetatePropyl AcetatePropylene Glycol Acetate2-Ethoxyethyl AcetateMethyl CellusolveBenzyl BenzoateIsopropyl MyristateTricresyl PhosphateHalogenated HydrocarbonsMethylene eon Carbon CyclohexanoneMethyl Ethyl KetoneIsopropylacetoneMethyl Isobutyl KetoneOrganic OxidesEthyl sulfoxide (DMSO)Isopropyl EtherMiscellaneousPhenol, Aqueous Solution, 10%Formaldehyde AqueousSolution, 30%Hydrogen Peroxide, 30%Silicone Oil/Mineral CCCCNDCNDC
Resourcesfor ChromatographersFor more information visit: www.thermoscientific.com/chromatography 2015 Thermo Fisher Scientific Inc. All rights reserved.Specifications, terms and pricing are subject to change. Not all products are available in all countries.Please consult your local sales representative for details.BR21473-EN 1215S
HyperSep Hypercarb SupelClean . Columns Application Notebook. LogP 3.6 Pk a 2.99. Thermo Scientific Chromatography Columns and Consumables 2016-2017 1-007 Sample Preparation Titan3 and Target2 Syringe Filters Sample preparation with Thermo Scientific Titan3 and Thermo Scientific Target2 syringe filters help provide consistent and reliable experimental results . Both these .
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