Section Contents - Anchem Plus

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SPE Column Selection . . . . . . . . . . 1-002SPE Procedure . . . . . . . . . . . . . . . . 1-005SOLA SPE Cartridges and Plates . . 1-007SPE Cartridges and Plates . . . . . . . 1-012Automated SPE . . . . . . . . . . . . . . . 1-048SPE Accessories. . . . . . . . . . . . . . . 1-051eVol XR Dispensing System. . . . . . 1-062QuEChERS Products . . . . . . . . . . . . 1-065Syringe Filters . . . . . . . . . . . . . . . . 1-072SOLA SPESyringe FiltersMaximizing reproducibilityand throughput for bio-analysisReliable elimination of particlesand micro organismsPAGE 1-007PAGE 1-072QuEChERS –Dispersive SPEEfficient pesticide samplepreparation from food matricesFeatured ProductsFeatured ProductsSample Preparation Section ContentsPAGE 1-065SLEHyperSep SPEProtein PrecipitationFast and effective sampleclean up of biological samplesReproducible and cleansample extractionFast removal of proteinsfrom biological samplesPAGE 1-035PAGE 1-012PAGE 1-034www.thermoscientific.com/chromatography1-001

Sample PreparationThermo Scientific SPE ProductsSPE 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 SampleSeparationModeRecommendedHyperSep 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 AXThermo Scientific Chromatography Columns and Consumables 2014-2015SOLA HRP, C18, C8,Phenyl, Retain PEP1-002Silica, AminopropylCyano, DiolSilica, Florisil, AminopropylCyano, DiolSCXSAXAminopropylVerify CXVerify AXVisit our online product selector tool atwww.thermoscientific.com/crc

Thermo Scientific HyperSep ProductAlternative ToHyperSep Retain CXPage 1-013Oasis MCX (Waters)Strata-X-C (Phenomenex)Bond Elut Plexa PCX (Agilent)StyreScreen DBX (UCT)H2 O-philic SC-DVB (JT Baker)HyperSep Retain AXPage 1-014Oasis MAX (Waters)Bond Elut Plexa PAX (Agilent)StyreScreen QAX (UCT)H2 O-philic SA-DVB (JT Baker)HyperSep HypercarbPage 1-015SupelClean ENVI-Carb Bond Elut CarbonHyperSep C18Page 1-022CLEAN-UP C18-USupelclean ENVI-18 / LC-18BAKERBOND SPE Polar Plus ISOLUTE C18CHROMABOND C18Bond Elut C18Strata C18-USep-Pak C18HyperSep C8Page 1-023HyperSep PhenylPage 1-024HyperSep SilicaPage 1-025CLEAN-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 SiAlternative ToHyperSep SCXPage 1-026CLEAN-UP BCXSupelclean LC-SCXBAKERBOND SPE Aromatic Sulfonic AcidISOLUTE SCXCHROMABOND SABond Elut SCXStrata SCXHyperSep SAXPage 1-027CLEAN-UP QAXSupelclean LC-SAXBAKERBOND SPE Quaternary amineISOLUTE SAXCHROMABOND SBBond Elut SAXStrata SAXHyperSep Verify CXPage 1-028CLEAN-UP DAUDiscovery DSC-MCAXISOLUTE HCXCHROMABOND DrugBond Elut Certify IStrata Screen-CHyperSep Verify AXPage 1-029CLEAN-UP THCISOLUTE HAXBond Elut Certify IIStrata Screen-AHyperSep FlorisilPage 1-030CLEAN-UP FlorisilSupelclean ENVI-Florisil / LC-FlorisilBAKERBOND SPE FlorisilISOLUTE FlorisilCHROMABOND FlorisilBond Elut FlorisilStrata FL-PRSep-Pak FlorisilHyperSep AminopropylPage 1-031CLEAN-UP AminopropylSupelclean LC-NH 2BAKERBOND SPE AminoISOLUTE NH2CHROMABOND NH 2Bond Elut NH2Strata NH2Sep-Pak NH2HyperSep CyanoPage 1-032Supelclean LC-CNBAKERBOND SPE CyanoCHROMABOND CNBond Elut CyanoSep-Pak CyanopropylStrata CNHyperSep DiolPage 1-033Supelclean LC-DiolBAKERBOND SPE DiolCHROMABOND OHBond Elut DiolSep-Pak Diolwww.thermoscientific.com/chromatographySPE Column SelectionHyperSep Retain PEPPage 1-012Oasis 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)Thermo ScientificHyperSep ProductSample Preparation SPE Phase Selection by Manufacturer1-003

Sample PreparationRemoving Uncertainty by Applying the Science to SPEOur 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 SPEThermo Scientific Chromatography Columns and Consumables 2014-2015Reducing 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:1-004Maximize detection selectivity Reduce ion suppression Reduce protein binding Reduce matrix interferences by eliminationof matrix and particulates Compatibility of solvent with analyticaltechniqueImprove 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 processingMatrix 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.

1. Sample Pre-treatment3. Column Pre-equilibration5. Wash Away InterferencesEquilibrate with weakly eluting solvent toprepare the phase for sample addition.Remove impurities bound less strongly thanthe compounds of interest. 2. Column Conditioning Use the same solvent as for samplepre-treatmentPrevent sorbent drying during columnequilibration4. Sample Loading3. Column Pre-equilibration5. Wash Away Interferences6. Elute Compounds of Interest1. Sample Pre-treatmentIt is important to optimize the samplefor effective analyte retention. The followingshould be considered: 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 conditioningApply 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 errorSelect 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 propertiesSPE Procedure4. Sample Loading Analytes are retained on the sorbent. Sample Preparation SPE Procedure – Six Steps for a Clean ExtractIt is important to optimize the Wash andElution steps in order to obtain maximumlevels of recovery.Get your copy of our applicationnotebook, featuring over150 SPE scientific.com/chromatography1-005

Sample PreparationMethod 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 100%90%80%70%60%50%40%30%20%10%LoadBlankThermo Scientific Chromatography Columns and Consumables 2014-20151-006EluteWash1009080706050403020100By 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 Notebook.

Join the revolutionThermo Scientific SOLA cartridges and plates provide clean, highly reproducible sample extractswith lower elution volumes due to the unique and innovative frit-less SPE design.SOLA SPE products provide unparalleled performance characteristics compared to conventional SPE,phospholipid removal and protein precipitation products. This includes: Higher levels of reproducibilityLower elution volumesHigh extract cleanlinessThis increased performance gives higher confidence in analytical results and lowers cost withoutcompromising ease of use or requiring complex method development.Technical InformationThe following information highlights the advantages associated with SOLA SPE products overconventional loose packed SPE products.Improved Reproducibility and RecoverySOLA SPE Cartridges and PlatesSOLA SPE products provide reduced failure rates, higher analysis speeds and lower solvent requirementswhich are critical in today’s high throughput bio-analytical and clinical research laboratory environments.Sample Preparation Thermo Scientific SOLA SPE Cartridges and PlatesThe data in Figure 1 and Table 1 shows that SOLA SPE products out perform conventional loose packedcompetitor SPE products even when utilizing the competitor recommended generic methodology.The information presented shows lower variability sample to sample, and higher recovery levels forSOLA SPE products compared to conventional loose packed competitor SPE products, ensuring you achievethe correct result time after time.100%SOLA SPECompetitor (i)Competitor (ii)% 200µL MeOH200µL Water1mL Sample200µL 5% MeOH in Water200µL MeOHFigure 1: SOLA SPE shows significantly higher reproducibility and recovery sionRecovery(%RSD n 30)Competitor (i)PrecisionRecovery(%RSD n 30)Competitor (ii)PrecisionRecovery(%RSD n 1%7.79%Table 1: SOLA SPE provides higher reproducibility and recovery levelswww.thermoscientific.com/chromatography1-007

Figure 2 shows that SOLA SPE products have consistent recoveries (across all 30 test samples). Theconventional loose packed SPE product from competitor (i) shows that on average one in every foursamples gives a significantly lower recovery. This results in inconsistencies in results which potentiallywould result in costly re-analysis. In comparison SOLA SPE products provide significantly higher levelsof reproducibility which is vitally important for high throughput studies.120CaffeineSOLA SPECompetitor (i)100% RecoverySample PreparationImproved reproducibility80604020012005101520Sample number253035101520Sample number253035101520Sample number253035Hydrocortisone% Recovery10080604020012005Carbamazepine% Recovery10080604020005Thermo Scientific Chromatography Columns and Consumables 2014-2015Figure 2: Shows inconsistency of loose packed products compared to SOLA SPE.1-008View product informationand application notes

Figure 3 shows that SOLA SPE products achieve excellent recovery levels even with low volumes ofextract solvents, resulting in a more concentrated analyte and increased sensitivity. Additional cost andtime saving benefits can be achieved from reduced sample dry down time and solvent usage. These lowvolume extractions would be significantly compromised when using a conventional loose packed,low bed weight, SPE product.Caffeine100SOLA SPECompetitor (i)Competitor (ii)% Recovery8060400150µL-20200µLSOLA SPE Cartridges and Plates20250µLElution VolumeHydrocortisone100% Recovery806040200150µL-20100200µLSample Preparation Higher sensitivity and lower solvent consumption250µLElution VolumeCarbamazepine% Recovery806040200-20150µL200µL250µLElution VolumeFigure 3: SOLA SPE products recovery and reproducibility at low extraction 9

SOLA SPE products proprietary manufacturing process provides a cleaner product and, as a result,a cleaner sample extract. This is shown in Figure 4 where SOLA SPE products are compared againstcompetitor (i) conventional loose packed SPE product, which have both been extracted withacetonitrile, dichloromethane and methanol, respectively.1009080Relative AbundanceSample PreparationCleanliness of Extract70605040SOLA SPECompetitor (i)30201000.00.51.01.52.02.53.0Methanol extract comparison:SOLA products versus competitor (i)3.54.04.5Time (minutes)5.05.56.06.57.010090Relative Abundance807060504030SOLA SPECompetitor (i)201000.00.51.01.52.02.53.0Acetonitrile extract comparison:SOLA products versus competitor (i)3.54.04.5Time (minutes)5.05.56.06.57.0100901-010Relative AbundanceThermo Scientific Chromatography Columns and Consumables 2014-20158070605040SOLA SPECompetitor (i)30201000.00.51.01.52.02.53.0Dichloromethane extract comparison:SOLA products versus competitor (i)3.54.04.5Time (minutes)5.05.56.06.57.0Figure 4: Shows SOLA SPE products are significantly cleaner than the equivalent loose packed SPE product from competitor (i)In today’s demanding bio-analytical or clinical research laboratory environment where reproducibility,certainty of results and cost saving are fundamental requirements, SOLA SPE products are anindispensible tool to provide confidence and first time/every time success in the analytical process.

SOLA SPE cartridges and plates are available in 10mg/1mL cartridge and 10mg/2mL 96 well plate formats.SOLA SPE CartridgesDescriptionSOLA HRPSOLA SCXSOLA SAXSOLA WCXSOLA WAXBed Weight (mg)1010101010Cat antity100 Pack100 Pack100 Pack100 Pack100 PackWell Volume (mL)22222Cat antity1 Pack1 Pack1 Pack1 Pack1 PackSOLA 96 Well PlatesDescriptionSOLA HRPSOLA SCXSOLA SAXSOLA WCXSOLA WAXBed Weight (mg)1010101010To learn more about how SOLA SPE can benefit your workflow,method scale down, applications, testimonials and more ientific.com/chromatographySOLA SPE Cartridges and PlatesColumn Volume (mL)11111Sample Preparation Product Information1-011

Sample PreparationThermo Scientific HyperSep SPE Cartridges and PlatesHyperSep Retain PEP SPE Cartridges and PlatesVersatile polymeric material for retention of polar and non-polar analytes Exceptional recoveries for polar and non-polar analytes High and consistent recoveries High capacity, high purity, highly porous polystyrene DVB material modifiedwith urea functional groups to give balanced retention of polar and non-polar analytes Fast and easy sample preparation and method development pH stable 0 to 14HyperSep Retain PEP SPE CartridgesBed Weight lumn Volume (mL)1336366363625625Cat. 0660107-21560107-21860107-214Quantity100 Pack50 Pack50 Pack30 Pack50 Pack30 Pack30 Pack50 Pack30 Pack50 Pack30 Pack20 Pack30 Pack20 PackNote: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.Thermo Scientific Chromatography Columns and Consumables 2014-2015HyperSep Retain PEP 96 Fixed Well Plates1-012Bed Weight (mg)3060Well Volume (mL)11Cat. No.60306-20760306-208Quantity1 Each1 EachNote: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.HyperSep Retain PEP 96 Removable Well PlateBed Weight (mg)30Volume (mL)1Cat. No.60303-207Note: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.Quantity1 EachApplications: Drugs and metabolitesin biological fluids Peptides in serum, plasmaor biological fluids Environmental samples

Sample Preparation HyperSep Retain CX SPE Cartridges and PlatesVersatile polymeric material for retention of basic compounds Exceptional recoveries for basic analytes High and consistent recoveries High capacity, high purity, highly porous polystyrene DVB material partially modifiedwith sulfonic acid functional groups Fast and easy sample preparation and method development pH stable 0 to 14Bed Weight (mg)303060601001502002005005001,0002,000Column Volume (mL)13366636362525Cat. 1560107-312Quantity100 Pack50 Pack50 Pack30 Pack30 Pack30 Pack50 Pack30 Pack50 Pack30 Pack20 Pack20 PackApplications: Analysis of a wide rangeof drugs of abuse including basicand neutral drugsHyperSep SPE Cartridges and PlatesHyperSep Retain CX SPE CartridgesNote: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.HyperSep Retain CX 96 Fixed Well PlatesBed Weight (mg)3060Well Volume (mL)11Cat. No.60306-30360306-304Quantity1 Each1 EachNote: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.HyperSep Retain CX 96 Removable Well PlateBed Weight (mg)30Volume (mL)1Cat. No.60303-307Quantity1 EachNote: For bed weights less than 30mg please refer to SOLA SPE products on page 1-011.www.thermoscientific.com/chromatography1-013

Sample PreparationHyperS

Chromatography Columns and Consumables 2014-2015 1-002 Sample Preparation SPE Cartridge Selection Sample Matrix Solubility of Sample Polarity of Sample Separation Mode Recommended HyperSep Sorbent(s) SOLA HRP, C18, C8, Phenyl, Retain PEP SOLA HRP, C18, C8, Retain PEP SOLA HRP, Hypercarb Retain PEP SOLA SCX/WCX, Verify CX Retain CX SCX Verify CX .

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