Thermo Scientific Chromatography Columns and Consumables

Thermo Scientific Chromatography Columns and Consumables 2014-2015

2014-2015

Your world of chromatography 4 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC ColumnsandAccessories and equipmentforfastLC,analyticalHPLCbiomolecule separations. accessories columns, innovative of range showcase ourlatestandmostcomprehensive 35 years,youcanrelyonthequalityofThermoScientific HPLCproducts. Herewe over for packing column and carbon manufacturing, bonded phase production As aleaderinLCcolumntechnologyincludingsilica,polymer andporousgraphitic predictability LC Columns and Accessories

Section Contents

Introduction ...... 4-001 LC Accessories...... 4-158 Column Selection...... 4-002 LC Equipment ...... 4-186 Column Protection ...... 4-016 LC Reagents ...... 4-193 Columns for Fast and Analytical LC . . 4-022 LC and LC-MS Instrument Parts . . 4-209 Columns for Biomolecules ...... 4-110 Technical Information...... 4-231

Featured Products

Accucore Columns Syncronis Columns Hypersil GOLD Columns Ultimate core performance - Consistent, reproducible separations, Outstanding peak shape for speed and selectivity combined column after column, time after time your separations

PAGE 4-042 PAGE 4-074 PAGE 4-024

MAbPac Columns Acclaim PepMap Columns Viper Connectors Unrivalled resolution and ef ciency The standard for peptide Simple, dead volume free plumbing in the analysis of protein variants separations in proteomics of HPLC and UHPLC systems

PAGE 4-126 PAGE 4-149 PAGE 4-158

www.thermoscienti c.com/chromatography 4-001 4 -002 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Common HPLC Phases and Uses HPLC Their Common outside ofwhichcolumnlifetimeswilldiminish. recommended pHRangeforthecolumnisillustrated, comparable retentionandresolution.The a needforhigheraqueousmobilephasestoachieve content inthemobilephase.Alowervalueindicates of neutralcompoundsandthehigherorganic higher thehydrophobicity, thegreaterretention the relativeretentiononcolumn.Generally, the The AUGwillshowyouHydrophobicitywhichgives column selection. for morehelpandinformationongeneralpurpose corresponding productpage(illustratedtotheright) Refer totheAdvancedUserGraphic(AUG)on Scientific alternativesforotherpopularcolumns. of ThermoScientificphaseswithspecifications,aswellrecommended Thermo properties, LC-MS requirementsorUSPspecifications. You will also findausefultable appropriate HPLCcolumnforyourapplication,basedonstationary phaseuse,analyte Information inthefollowingsectionwillhelpyoumakeaninformed decisiononthe LC ColumnSelection carbon graphitic Porous Anion Strong AX SAX Diol rophenyl Penta uo- APS Amino CN CPS, Cyano C18 Butyl C8 SAS C4 C1 Silica Silica Silica Silica Name Common SCX Strong Cation Cation Strong SCX Phenyl PGC (PEI) Polyethyleneimine Anion Exchanger Exchanger PFP ODS MOS Name Alternative Exchanger 100% carbon 100% -(CH -RN+(CH -(CH -C -(CH -(CH -C -C -C -(CH CH -OH Group Functional -RSO -(CH 18 6 8 4 2 F H H H (CH 2 5 2 2 3 3 2 17 9 CH )C ) ) ) 3 ) 37 3 3 3

2O H- H- NH CN CN

6 2 2 H OH) NH-) 3 ) 5 2

3

2 n Phase Normal

• • • • • • • • • • • the bestchoiceforyourapplication. Support andtapintoourexpertisetohelpmake see thebackcovertocontactourexpertTechnical For additionalhelpincolumnselection,please most appropriatestationaryphaseforyouranalysis. stationary phases,allowingyoutochoosequicklythe to quicklycomparethemaincharacteristicsofmultiple USP code.Thesegraphicsaredesignedtoallowyou will seetheparticlesizesavailable,aswell is relatedtothehydrophobicity. Belowtheicon,you proteins orpeptides.ThepercentageCarbonLoad being moreapplicabletolargeranalytessuchas The PoreSizeisshown,withlargerporesizecolumns Phase Reverse • • • • • • Exchange Ion • • • HILIC

• • diastereoisomers). isomers, (e.g., compounds similar structurally of separation columns; based silica conventional using dif are retain to cult that compounds polar highly of separation the for useful Particularly oligonucleotides. and nucleotides acids, Organic nucleosides. and nucleotides acids, Organic Organic bases. Normal Phase: Reversed Phase: and isomers. compounds polar halogenated, for retention and selectivity Extra polar and compounds moderately compounds. Aromatic Normal Phase: acids. organic and anions exchange: anion Weak compounds. polar other and analysis Carbohydrate HILIC: solutes. different very of mixtures for and applications pharmaceutical of range wide a for Useful C18 phases. and C8 the of that to different is selectivity the phase, reversed in used When separations. gradient phase normal for silica base than suitable more compounds, polar for selectivity Unique pharmaceuticals, steroids, acids, fatty phthalates, environmental etc. for widely Used phases. alkyl-bonded the of retentive Most proteins, pharmaceuticals, steroids, environmental samples. and peptides small for used normally C18; than retentive Less proteins. and peptides of Separation C18. C8, than retention Shorter compounds. multi-functional and polar moderately for used Typically solutes. non-polar for phases bonded group alkyl all of retentive Least compounds. organic polar moderately and Non-polar Application Alternative selectivity to silica. Good for aromatics. for Good silica. to selectivity Alternative Similar selectivity to silica, but less polar. less but silica, Similar to selectivity Proteins, peptides. Advanced UserGraphic(AUG) Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 02 01 0 Low 2.6µm 80Å 5 2 to8 USP High L11 300 5 4 HPLC Stationary Phase Column Selection LC Columns and Accessories Before beginning a new analysis, consider the physical and chemical properties of the analyte(s), the mode of analysis and how the analyte(s) will interact with the surface of the chromatographic phase. To aid column selection, the following guide may be useful.

Non-polar

Analyte Solubilty Analyte Chemical Properties Mode of Separation Recommended Phase

Acclaim Mixed-Mode WAX-1 4-069 Acidic pK > 2 Anion exchange / Reversed-phase mixed mode a Acclaim Trinity P1 4-091 Accucore C18 4-045 Accucore RP-MS 4-044 Accucore C8 4-046 Accucore C30 4-052 Accucore Phenyl-Hexyl 4-049 Acclaim 120 C18 4-061

Acclaim 120 C8 4-062 Non-polar Reversed phase >> Acclaim C30 4-064 Column Selection Acclaim Phenyl-1 4-063 Hypersil GOLD 4-026 Hypersil GOLD C8 4-027 Hypersil GOLD C4 4-028 Neutral Syncronis C18 4-075 Syncronis C8 4-076 Accucore aQ 4-047 Accucore Polar Premium 4-048 Accucore PFP 4-051 Accucore Phenyl-X 4-050 Acclaim PolarAdvantage 4-065 Polar Moderately-polar Reversed phase solvents Acclaim PolarAdvantage II 4-066 Hypersil GOLD aQ 4-029 Hypersil GOLD PFP 4-030 Hypersil GOLD Phenyl 4-031 Syncronis aQ 4-077 Syncronis Phenyl 4-078 Accucore C18 4-045 Accucore RP-MS 4-044 Accucore C8 4-046 Accucore C30 4-052 Accucore Phenyl-Hexyl 4-049 Acclaim 120 C18 4-061 Acclaim 120 C8 4-062 Reversed phase Acclaim C30 4-064 Basic pK < 10 a Acclaim Phenyl-1 4-063 Hypersil GOLD 4-026 Hypersil GOLD C8 4-027 Hypersil GOLD C4 4-028 Syncronis C18 4-075 Syncronis C8 4-076 Acclaim Mixed-Mode WCX-1 4-070 Cation exchange / Reversed-phase mixed mode Acclaim Trinity P1 4-091

www.thermoscienti c.com/chromatography 4-003 4 -004 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories separation ofbiomoleculescanbefoundinthetableonpage The abovetablerecommendscolumnsfortheseparationofsmallmoleculeanalytes. Columnrecommendationsforthe Phase Selection Column Stationary HPLC Polar Solubilty Analyte solvents Polar solvents Non-polar Analyte ChemicalProperties Neutral Basic Acidic pK pK a a >10 <2 4-111 Mode ofSeparation HILIC Polar retentioneffectongraphite HILIC Anion exchange Polar retentioneffectongraphite Normal phase HILIC Cation exchange Polar retentioneffectongraphite continued Hypercarb Hypersil GOLD AX GOLD Hypersil Hypersil GOLD SAX GOLD Hypersil Accucore Urea-HILIC Accucore Accucore 150-Amide-HILIC Accucore Acclaim HILIC-10 HILIC-10 Acclaim Syncronis HILIC HILIC Syncronis Hypercarb Accucore HILIC HILIC Accucore Acclaim Mixed-Mode HILIC-1 Mixed-Mode Acclaim Acclaim Mixed-Mode WAX-1 Mixed-Mode Acclaim Acclaim Mixed-Mode WCX-1 Acclaim Trinity P1 Trinity Acclaim Hypersil GOLD Amino GOLD Hypersil Hypersil GOLD HILIC GOLD Hypersil Syncronis Amino Amino Syncronis Syncronis HILIC HILIC Syncronis Hypercarb Acclaim Mixed-Mode WCX-1 Acclaim Trinity P1 Trinity Acclaim Accucore Urea-HILIC Accucore Accucore 150-Amide-HILIC Accucore Acclaim HILIC-10 HILIC-10 Acclaim Syncronis HILIC HILIC Syncronis Hypersil GOLD Amino GOLD Hypersil Hypersil GOLD CN GOLD Hypersil Hypersil GOLD Silica GOLD Hypersil Syncronis Amino Amino Syncronis Syncronis Silica Silica Syncronis Recommended Phase 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 -084 -034 -035 -054 -146 -067 -081 -084 -053 -068 -069 -070 -091 -033 -037 -079 -081 -084 -070 -091 -054 -146 -067 -081 -033 -032 -036 -079 -080 Column Format Selection for LC-MS LC Columns and Accessories The Thermo Scientific range offers a broad array of column designs and stationary phases optimized for LC-MS applications. Use the following table to help you choose your column format to best meet your application needs. A variety of HPLC column hardware configurations are available, designed to give superior results for high speed, high sensitivity, high efficiency and convenience. A wide range of particle and monolithic stationary phases allows choices for optimized selectivity.

Column Hardware Selection for LC-MS LC-MS Application Column Hardware Design Description High throughput analysis Javelin HTS columns Direct-connection columns Slim design, 20mm length, 1mm to 4.6mm ID High sensitivity analysis Acclaim PepMap nano, capillary Nano, capillary and micro columns 0.075mm to 1mm ID; 50 to 250mm length and micro columns nanoViper format offers ngertight dead volume free connection to 1000 bar Proteomics analysis EASY-Spray columns Combined column/emitter design with nanoViper connection. Heated  exible silica columns 50µm and 75µm ID; 150 to 500mm length Acclaim PepMap nano columns, nanoViper offers ngertight dead volume free connection to 1000 bar Accucore nano columns, nanoViper Flexible silica columns 50μm and 75μm ID; 50 to 500mm length Trap column 20mm x 100μm ID

Acclaim PepMap nano columns, Flexible silica columns 50μm and 75μm ID; 50 to 500mm length >>

classic Trap column 20mm x 100μm ID Column Selection EASY-Column Flexible silica columns 100mm x 75μm Trap column 20mm x 100μm ID PepSwift monolithic columns Flexible fused silica columns, nanoViper connections 100µm to 500µm ID, 50 to 250mm length Trap columns 5mm x 200µm ID

Various HPLC columns, throughout this LC section, can also be used for LC-MS application. Typical flow rates and MS source compatibility for these columns are shown in the table on the next page

www.thermoscienti c.com/chromatography 4-005 4 -006 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories ProSwift isalsoavailableinlargerIDsforhighthroughputapplications. Monolith columnsforLC-MS Packed columnselectionforLC-MS LC-MS for Selection Column nlt oeua egtColumnID(mm) Analyte MolecularWeight Weight Molecular Analyte > 1000Da < 1000Da > 1000Da < 1000Da Polarity Sample Low (or ionizable) High Medium Ionization Interface APCI ESI ESI ESI APCI Sensitivity Relative 1.0 0.1, 0.2,0.5 High Low High Low High Low High Low High Low (mm) Column ID 0.2 –0.05 1.0 –0.3 2.1, 1.0 0.2 –0.05 1.0 –0.3 2.1, 1.0 0.2 –0.05 1.0 –0.3 2.1, 1.0 2.1, 1.0 4.6, 4.0,3.0 2.1, 1.0 4.6, 4.0,3.0 continued lwRt µ/i)ColumnHardware Flow Rate(µL/min) 40 –200 0.7 –25 (µL/min) Flow Rate 200 –50 200 –50 200 –50 200 –50 2000 –200 200 –50 2000 –200 2 –0.2 50 –5 2 –0.2 50 –5 2 –0.2 50 –5 Column Hardware EASY-Spray column,EASY-Column, AcclaimPepMap(RSLC)nano,Accucorenano Acclaim PepMap(RSLC)capillaryandmicro Analytical, JavelinHTS EASY-Spray column,EASY-Column, AcclaimPepMap(RSLC)nano,Accucorenano Acclaim PepMap(RSLC)capillaryandmicro Analytical, JavelinHTS EASY-Spray column,EASY-Column, AcclaimPepMap(RSLC)nano,Accucorenano Acclaim PepMap(RSLC)capillaryandmicro Analytical, JavelinHTS Analytical, JavelinHTS Javelin HTS,Analytical Analytical, JavelinHTS Javelin HTS,Analytical ProSwift Monolith PepSwift Monolith HPLC Column Selection by U.S. Pharmacopeia Speci cations* LC Columns and Accessories

USP Code Description Recommended Phase Page L1 Octadecyl silane (C18) chemically bonded to porous Acclaim 120 C18 4-061 or ceramic microparticles, 1.5 to 10μm in diameter, Acclaim 300 C18 4-142 or a monolithic rod Accucore C18 4-045 Accucore aQ 4-047 Accucore 150-C18 4-144 Accucore XL C18 4-056 BioBasic 18 4-140 Hypersil GOLD 4-026 Hypersil GOLD aQ 4-029 Acclaim PepMap 100 C18 4-149 Syncronis C18 4-075 Syncronis aQ 4-077 L3 Porous silica particles, 1.5 to 10μm in diameter, Accucore HILIC 4-053

or a monolithic rod Hypersil GOLD Silica 4-036

Syncronis Silica 4-080 >>

L7 Octyl silane chemically bonded to totally Acclaim 120 C8 4-061 Column Selection or super cially porous silica particles, Accucore C8 4-046 1.5 to 10μm in diameter, or a monolithic rod Accucore XL C8 4-057 BioBasic 8 4-140 Hypersil GOLD C8 4-027 Acclaim PepMap 100 C8 4-150 Syncronis C8 4-076 L8 An essentially monomolecular layer of Hypersil GOLD Amino 4-033 aminopropylsilane chemically bonded to totally Syncronis Amino 4-079 porous silica gel support, 1.5 to 10μm in diameter L10 Nitrile groups chemically bonded to porous silica Hypersil GOLD CN 4-032 particles, 1.5 to 10μm in diameter L11 Phenyl groups chemically bonded to porous silica Accucore Phenyl-Hexyl 4-049 particles, 1.5 to 10μm in diameter Hypersil GOLD Phenyl 4-031 Syncronis Phenyl 4-078 L13 Trimethylsilane chemically bonded to porous silica BETASIL C1 Inquire particles, 3 to 10μm in diameter Hypersil SAS 4-107 L14 Silica gel having a chemically bonded, strongly basic Hypersil GOLD SAX 4-035 quaternary ammonium anion exchange coating, Hypersil SAX 4-109 5 to 10μm in diameter L15 Hexylsilane (C6) chemically bonded to totally porous BETASIL C6 Inquire silica particles, 3 to 10μm in diameter

www.thermoscienti c.com/chromatography 4-007 4 -008 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories foreachcategoryarealso available. * These aretherecommendedThermo ScientificHPLCcolumnsforvariousUSPcategoriesalthoughother HPLC Column Selection by U.S. Pharmacopeia U.S. Speci by Selection Column HPLC cations L34 L78 L50 L33 L62 L58 L46 L26 L60 L17 L52 L43 L20 L59 L19 USP Code lead form, 7 to 9μm in diameter in 9μm 7to form, lead the in copolymer styrene-divinylbenzene cross-linked sulfonated of consisting resin exchange cation Strong 1.0 to 50μm in diameter or a monolithic rod amonolithic or diameter in 1.0 50μm to micro-particles, ceramic or non-porous or porous to bonded chemically groups functional groups) amino quaternary or tertiary secondary, (primary, anion-exchange and C8) than longer chain alkyl (an reversed-phase both of consists that ligand A silane a surface area of not less than 350m than less not of area a surface and diameter, in 15µm 3to copolymer, divinylbenzene with cross-linked 55% ethylvinylbenzene, of consists anion-exchangestrong functionalities. resin The and retention reverse-phase with resin Multifunction stability pH provide to processed and silica-based, spherical, is It daltons. 500,000 to 4,000 of arange over size molecular by dextrans separate to capacity the having Packing silica, 3 to 15µm in diameter in 15µm 3to silica, spherical porous afully on phase bonded silane C30 sodium form, about 6 to 30μm in diameter in 30μm 6to about form, sodium the in copolymer styrene-divinylbenzene cross-linked sulfonated of consisting resin exchange cation Strong about 9 to 11um 9to about diameter in beads latex functionalized amine quaternary with agglomerated substrate Polystyrene/divinylbenzene diameter in 1.5 10μm to particles, silica porous totally to bonded chemically silane Butyl with alkyl amide groups and endcapped and groups amide alkyl with modi covalently ed been has which of surface the diameter, in less or 10μm gel, silica porous Spherical, hydrogen form, 6 to 12μm in diameter in 12μm 6to form, hydrogen the in copolymer styrene-divinylbenzene cross-linked sulfonated of consisting resin exchange cation Strong with sulfopropyl groups 5 to 10μm in diameter in 10μm 5to groups sulfopropyl with silica porous of made resin exchange cation A strong particles by a propyl spacer, 1.5 to 10μm in diameter in 1.5 10μm to spacer, apropyl by particles Penta groups chemicallyuorophenyl to bonded silica diameter in 1.5 10μm to particles, hybrid or silica porous to bonded chemically groups Dihydroxypropane silica or hybrid packing with a hydrophilic coating ahydrophilic with packing hybrid or silica 1.5 10um, to spherical, is packing The kDa. 7,000 5 to of range the over weight) molecular by (separation proteins of separation exclusion size the for Packing calcium form, about 9μm in diameter in 9μm about form, calcium the in copolymer styrene-divinylbenzene cross-linked sulfonated of consisting resin exchange cation Strong Description cross-linked with divinylbenzene styrene of consisting particles latex functionalized ammonium quaternary with coated is Substrate 2 per g. g. per HyperREZ XP Carbohydrate Pb Carbohydrate XP HyperREZ Acclaim Mixed-Mode WAX-1 Mixed-Mode Acclaim MAbPac SEC-1 1000 SEC BioBasic 300 SEC BioBasic 120 SEC BioBasic Accucore C30 Accucore C30 Acclaim HyperREZ XP Carbohydrate Na Carbohydrate XP HyperREZ OmniPac PAX-100 OmniPac C4 300 PepMap Acclaim C4 GOLD Hypersil 4 BioBasic 150-C4 Accucore Accucore Polar Premium Polar Accucore II PolarAdvantage Acclaim PolarAdvantage Acclaim HyperREZ XP Carbohydrate H Carbohydrate XP HyperREZ BioBasic SCX BioBasic Hypersil GOLD PFP PFP GOLD Hypersil PFP Accucore Diol BETASIL MAbPac SEC-1 (5μm) 300 SEC BioBasic Ca Carbohydrate XP HyperREZ Recommended Phase Recommended OmniPac PAX-500 OmniPac HyperREZ XP Sugar Alcohols Sugar XP HyperREZ HyperREZ XP Organic Acids Organic XP HyperREZ 2+ 2+ + +

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Inquire 4 4 Page 4 4 4 4 4 4 4 -069 -129 -139 -139 -139 -052 -064 -103 -149 -028 -140 -145 -048 -066 -065 -141 -030 -051 -129 -139 -103 -156 -156 -156 -156 -156 -156 *

continued Thermo Scienti c HPLC Phases LC Columns and Accessories The tables below list Thermo Scientific HPLC sorbents offered for small molecules separations. For Thermo Scientific phases for the separations of biomolecules see page 4-112. Please also refer to the Advanced User Graphic (AUG) for each HPLC phase on the pages indicated.

Particle Size Pore Nominal Surface % USP Phase Phase Particle Type Endcapping Page (µm) Size (Å) Area (m2/g) Carbon Code Code Acclaim Phases 120 C18 Spherical, fully porous silica 2.2, 3, 5 120 300 18 Yes L1 – 4-061 300 C18 Spherical, fully porous silica 3 300 100 8 Yes L1 – 4-142 120 C8 Spherical, fully porous silica 2.2, 3, 5 120 300 11 Yes L7 – 4-062 Phenyl-1 Spherical, fully porous silica 3 120 300 13 Yes L11 – 4-063 C30 Spherical, fully porous silica 3, 5 200 200 13 Proprietary L62 – 4-064 PA Spherical, fully porous silica 2.2, 3, 5 120 300 16 Yes L60 – 4-065 PA II Spherical, fully porous silica 2.2, 3, 5 120 300 16 Yes L60 – 4-066 HILIC-10 Spherical, fully porous silica 3 120 300 8 Yes –– 4-067

Trinity P1 Nano polymer silica hybrid 3 300 100 – Proprietary –– 4-091 Trinity P2 Nano polymer silica hybrid 3 300 100 – Proprietary –– 4-092 >>

Trinity Q1 Nano polymer silica hybrid 3 300 300 – Proprietary –– 4-099 Column Selection Mixed Mode HILIC-1 Spherical, fully porous silica 3 120 300 – Proprietary –– 4-068 Mixed Mode WAX-1 Spherical, fully porous silica 3, 5 120 300 – Proprietary –– 4-069 Mixed Mode WCX-1 Spherical, fully porous silica 3, 5 120 300 – Proprietary L78 – 4-070 Organic Acid Spherical, fully porous silica 3, 5 120 300 – Yes –– 4-093 Surfactant Spherical, fully porous silica 3, 5 120 300 – Yes –– 4-094 Surfactant Plus Spherical, fully porous silica 3, 5 120 300 – Yes –– 4-095 Explosives Spherical, fully porous silica 3, 5 120 300 – Yes –– 4-097 Carbamate Spherical, fully porous silica 3, 5 120 300 – Yes –– 4-100 Carbonyl C18 Spherical, fully porous silica 2.2 120 300 – Yes –– 4-101 SEC Spherical, resin 5, 7 300 1000 –––– 4-088 Accucore Phases RP-MS Spherical, solid core silica 2.6 80 130 7 Yes – 176 4-044 C18 Spherical, solid core silica 2.6 80 130 9 Yes L1 171 4-045 C8 Spherical, solid core silica 2.6 80 130 5 Yes L7 172 4-046 aQ Spherical, solid core silica 2.6 80 130 9 Polar L1 173 4-047 Polar Premium Spherical, solid core silica 2.6 150 90 8 Yes L60 280 4-048 Phenyl-Hexyl Spherical, solid core silica 2.6 80 130 5 Yes L11 179 4-049 Phenyl-X Spherical, solid core silica 2.6 80 130 6 Yes – 279 4-050 PFP Spherical, solid core silica 2.6 80 130 5 Yes L43 174 4-051 C30 Spherical, solid core silica 2.6 150 90 5 Yes L62 278 4-052 HILIC Spherical, solid core silica 2.6 80 130 –– L3 175 4-053 Urea-HILIC Spherical, solid core silica 2.6 80 130 ––– 277 4-054 Accucore XL Phases C18 Spherical, solid core silica 4 80 90 7 Yes L1 741 4-056 C8 Spherical, solid core silica 4 80 90 4 Yes L7 742 4-057

www.thermoscienti c.com/chromatography 4-009 4 -010 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Thermo Scienti Thermo Phases c HPLC Amino Phenyl aQ C8 C18 Syncronis Phases PAH Green Phase Hypersil HILIC Silica SAX AX Amino CN (Cyano) CN Phenyl PFP aQ C4 C8 C18 selectivity Phases GOLD Hypersil Cyano Phenyl C8 C18 BDS Phases Hypersil SAX Silica APS-2 CPS-2 CPS Phenyl-2 Phenyl SAS (C1) SAS MOS-2 (C8) MOS-2 MOS (C8) MOS ODS-2 (C18) HILIC ODS (C18) Phases Hypersil Silica Phase Hypercarb Phase Hypercarb Spherical, fully porous silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, Spherical, fully porous silica porous fully Spherical, Spherical, fully porous silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, peia,flypru iia1.9, 5 3, silica porous fully Spherical, Spherical, fully porous silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, Spherical, fully porous silica 2.4, 3, 5 3, 2.4, silica porous fully Spherical, Spherical, fully porous silica 2.4, 3, 5 3, 2.4, silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia1.7, 5 3, silica porous fully Spherical, Particle Type Particle Spherical, porous graphitic carbon graphitic porous Spherical, 1.9, 5 3, 1.9, 5 3, 1.9, 12 5, 3, (µm) Size Particle 1.7, 5 3, 3, 5 3, continued 100 100 100 100 120 175 175 175 175 175 175 175 175 175 175 175 175 130 130 130 130 120 120 120 120 120 120 120 120 120 120 80 100 120 100 Size (Å) Size Pore 10 250 0 320 320 320 320 170 220 220 220 220 220 220 220 220 220 220 220 220 170 170 170 170 170 170 170 170 170 170 170 170 170 170 220 320 170 320 Area (m Nominal Surface 120 320 2 /g) 11 19 10 16 13.5 6 – – 2.5 6 2 4 8.5 8 12 5 8 10 4 5 7 11 2.5 – – 1.9 4 4 5 5 2.5 6.5 6.5 11 – – – 5 10 – – 100 4 Carbon % Yes Polar Yes Yes Yes No Yes No Yes Yes Yes Yes Yes Polar Yes Yes Yes Yes Yes Yes Yes Yes No Yes No Yes No Yes Yes No Yes Yes – – Yes Endcapping L11 L1 L7 L1 – – L3 L14 – L8 L10 L11 L43 L1 L26 L7 L1 L10 L11 L7 L1 L14 L3 L8 L10 L10 L11 L11 L13 L7 L7 L1 L1 L3 L8 Code USP 979 973 972 971 311 265 251 263 261 257 258 259 259 254 253 255 252 250 288 289 282 281 341 300 307 318 308 319 309 305 303 302 316 975 301 970 350 977 Code Phase 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Page -078 -077 -076 -075 -102 -037 -036 -035 -034 -033 -032 -031 -030 -029 -028 -027 -026 -105 -105 -105 -105 -109 -109 -108 -108 -108 -108 -108 -107 -107 -107 -107 -081 -106 -080 -084 -079 HPLC Column Selection by Manufacturer LC Columns and Accessories

To find a suitable Thermo Scientific alternative provide the same retention or selectivity, but The following table is not complete in terms to another manufacturer’s columns, refer to the should be suitably similar in character to allow of manufacturer or products offered. Although selection guide below. The Thermo Scientific a similar or improved separation to be achieved every effort is made to ensure that the product alternative phases are selected based on a with some method optimization. The user should information provided is as accurate as possible, combination of physical and chemical refer to the individual phase information to some errors may occur in collation and transcription. similarities as well as mode of retention. ensure that the characteristics of the alternative We cannot accept any responsibility for the use These alternatives are not guaranteed to match the requirements of their separation. of the following information.

Phase Manufacturer Pore Size (Å) Area (m2/g) % Carbon Recommended Thermo Scienti c Alternative Page ACE C18 ACT 100 300 15.5 Syncronis C18 4-075 ACE C8 ACT 100 300 9 Syncronis C8 4-076 ACE Phenyl ACT 100 300 9.5 Syncronis Phenyl 4-078 ACE AQ ACT 100 300 14 Syncronis aQ 4-077 ACE C18-300 ACT 300 100 9 BioBasic 18 4-140

ACE C8-300 ACT 300 100 5 BioBasic 8 4-140

ACE C4-300 ACT 300 100 2.6 BioBasic 4 4-140 >> ™

ACQUITY UPLC BEH HILIC Waters 130 185 – Hypersil GOLD Silica (1.9µm) 4-036 Column Selection ACQUITY UPLC HSS C18 Waters 100 230 15 Hypersil GOLD (1.9µm) 4-026 ACQUITY UPLC BEH C18 Waters 130 185 18 Hypersil GOLD (1.9µm) 4-026 ACQUITY UPLC BEH C8 Waters 130 185 13 Hypersil GOLD C8 (1.9µm) 4-027 ACQUITY UPLC BEH Phenyl Waters 130 185 15 Hypersil GOLD Phenyl (1.9µm) 4-031 ACQUITY UPLC HSS T3 Waters 100 230 11 Hypersil GOLD aQ (1.9µm) 4-029 Aeris PEPTIDE XB-C18 Phenomenex 100 200 – Accucore 150-C18 4-144 Aeris WIDEPORE XB-C18 Phenomenex 200 25 – Accucore 150-C18 4-144 Aeris WIDEPORE XB-C8 Phenomenex 200 25 – Accuccore 150-C4 4-145 Aeris WIDEPORE XB-C4 Phenomenex 200 25 – Accuccore 150-C4 4-145 Alltima™ HP C18 Grace 190 200 12 Hypersil GOLD 4-026 Alltima HP C18 AQ Grace 100 450 20 Hypersil GOLD aQ 4-029 Alltima HP C18 HiLoad Grace 100 450 24 Syncronis C18 4-075 Alltima HP C8 Grace 190 200 8 Hypersil GOLD C8 4-027 Alltima HP CN Grace 190 200 4 Hypersil GOLD CN 4-032 Alltima HP Silica Grace 190 200 – Hypersil GOLD Silica 4-036 Aminex™ HPX42C Bio-Rad – – – HyperREZ XP Carbohydrate Ca2+ 4-156 Aminex HPX72S Bio-Rad – – – HyperREZ XP Carbohydrate H+ 4-156 Aminex HPX87C Bio-Rad – – – HyperREZ XP Carbohydrate Ca2+ 4-156 Aminex HPX87H Bio-Rad – – – HyperREZ XP Carbohydrate H+ 4-156 Aminex HPX87N Bio-Rad – – – HyperREZ XP Carbohydrate Na+ 4-156 Aminex HPX87P Bio-Rad – – – HyperREZ XP Carbohydrate Pb2+ 4-156 AQUA™ C18 Phenomenex 125 320 15 Hypersil GOLD aQ 4-029 Ascentis™ C18 Supelco 100 450 25 Syncronis C18 4-075 Ascentis C8 Supelco 100 450 15 Syncronis C8 4-076 Ascentis Express C18 Supelco 90 150 – Accucore C18 4-045 Ascentis Express C8 Supelco 90 150 – Accucore C8 4-046 Ascentis Express F5 Supelco 90 150 – Accucore PFP 4-051 Ascentis Express HILIC Supelco 90 150 – Accucore HILIC 4-053 Ascentis Express Supelco 90 150 – Accucore Phenyl-Hexyl 4-049 Phenyl-Hexyl Ascentis Express Supelco 90 150 – Accucore Polar Premium 4-048 RP-Amide Ascentis Express Peptide Supelco 160 80 – Accucore 150-C18 4-144 ES-C18 Ascentis Phenyl Supelco 100 450 19 Syncronis Phenyl 4-078

www.thermoscienti c.com/chromatography 4-011 4 -012 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories HPLC Column Selection by Manufacturer by Selection Column HPLC Luna PFP (2) PFP Luna NH2 Luna Luna HILIC CN Luna (2) C8 Luna LiChrospher Silica Kromasil C4 Kromasil C18 Kromasil XB-C18 Kinetex iee hnlHxlPhenomenex Kinetex Phenyl-Hexyl PFP Kinetex Kinetex HILIC C8 Kinetex C18 Kinetex C4 Jupiter Inertsil ES-C18 Peptide HALO Halo RP-Amide Halo Phenyl-Hexyl PFP Halo HILIC Halo C8 Halo C18 Halo Gemini Discovery Cyano Discovery C8 C18 Discovery C8 Pore Wide BIO Discovery LiChrospher NH LiChrospher Diol LiChrospher Luna ihope P8 Merck RP-8e LiChrospher ihope P8Merck RP-8 LiChrospher ihope P1eMerck RP-18e LiChrospher 18 RP LiChrospher Atlantis Jupiter M80 J’Sphere Silica Inertsil Inertsil Phenyl Inertsil ODS3V C8 Inertsil Capcell Pak C18 AQ Pak Capcell C18 Core Capcell Atlantis dC18 Atlantis HILICSilica Atlantis T3 Discovery HILIC Cortecs C18+ Cortecs C18 Cortecs Phase ™ C18 (2) C18 ™ ™ ™ ™ C18 C4 C18 ™ dC18 BIO Wide Pore C18 Pore Wide BIO ™ CN 2 Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Akzo-Nobel Akzo-Nobel Akzo-Nobel Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex AMT AMT AMT AMT AMT AMT AMT Phenomenex Supelco Supelco Supelco Supelco Supelco Merck Merck Merck Merck YMC GL Sciences GL Sciences GL Sciences GL Sciences GL Sciences Shiseido Shiseido Shiseido Waters Waters Waters Waters Waters Waters Waters Manufacturer 100 100 200 100 100 100 100 100 100 100 100 100 100 100 100 300 300 160 90 90 90 90 90 90 110 180 180 180 300 300 100 100 100 100 100 100 100 80 150 150 100 150 150 120 90 100 100 100 100 90 90 90 (Å) Size Pore 400 400 200 400 400 400 340 340 340 – – – – – – 170 170 80 150 150 150 150 150 150 375 200 200 200 – – 350 350 350 350 350 350 350 – 320 320 450 320 320 300 150 330 300 300 330 100 100 100 Area (m 2 continued /g) – Hypersil GOLD CN GOLD Hypersil 5.7 10 5.7 – 14 18 8 Hypersil GOLD C4 GOLD Hypersil – 8 19 12 – 9 Accucore PFP Accucore 9 – 10 12 5 13 – Accucore Polar Premium Polar Accucore PFP Accucore – – C8 Accucore – – – – – 14 – Hypersil GOLD CN GOLD Hypersil C8 GOLD Hypersil – – 14 – – 7 Hypersil GOLD CN GOLD Hypersil – 7 13 13 22 21 5 % Carbon 14 – C4 GOLD Hypersil 10 15 11 8 11 7 12 – 14 12 – 5.7 6.6 Hypersil GOLD PFP GOLD Hypersil Amino Syncronis Diol BETASIL C8 Syncronis C18 Syncronis Syncronis Silica Syncronis C18 Syncronis C18 Accucore Phenyl-Hexyl Accucore Accucore HILIC Accucore C8 Accucore C18 Accucore C4 BioBasic 18 BioBasic Accucore 150-C18 Accucore Phenyl-Hexyl Accucore HILIC Accucore C18 Accucore Hypersil GOLD Hypersil GOLD 8 BioBasic 18 BioBasic BETASIL Diol BETASIL Syncronis C8 Syncronis Syncronis C8 Syncronis Syncronis C18 Syncronis Syncronis C18 Syncronis Amino Syncronis Recommended Thermo Scienti Recommended Thermo c Alternative Acclaim PolarAdvantage II PolarAdvantage Acclaim Silica Syncronis Phenyl Syncronis C18 Syncronis C8 Syncronis Acclaim Polar Advantage II Advantage Polar Acclaim C18 Accucore aQ GOLD Hypersil Silica GOLD Hypersil Hypersil GOLD II Advantage Polar Acclaim Accucore HILIC Accucore Premium Polar Accucore C18 Accucore 4 4 Inquire 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Inquire 4 4 4 4 4 4 Page 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 -030 -079 -076 -075 -032 -080 -075 -045 -049 -028 -053 -046 -045 -140 -140 -051 -144 -049 -053 -045 -026 -048 -051 -046 -026 -140 -140 -032 -027 -032 -076 -076 -075 -075 -079 -066 -080 -078 -075 -076 -028 -066 -045 -029 -036 -026 -066 -053 -048 -045 LC Columns and Accessories

Phase Manufacturer Pore Size (Å) Area (m2/g) % Carbon Recommended Thermo Scienti c Alternative Page Luna SCX Phenomenex 100 400 – BioBasic SCX 4-141 Luna Silica (2) Phenomenex 100 400 – Syncronis Silica 4-080 µBondapak™ C18 Waters 125 330 10 Hypersil GOLD 4-026 µBondapak CN Waters 125 330 – Hypersil GOLD CN 4-032

µBondapak NH2 Waters 125 330 4 Hypersil APS-2 4-108 µBondapak Phenyl Waters 125 330 – Hypersil GOLD Phenyl 4-031 Nova-Pak™ (HR) C18 Waters 60 120 7 Hypersil GOLD 4-026 Nova-Pak C8 Waters 60 120 – Hypersil GOLD C8 4-027 Nova-Pak CN Waters 60 120 – Hypersil GOLD CN 4-032 Nova-Pak Phenyl Waters 60 120 5 Hypersil GOLD Phenyl 4-031 Nova-Pak Silica Waters 60 120 – Hypersil GOLD Silica 4-036 NUCLEODUR™ C18 EC Macherey-Nagel 110 340 18 Syncronis C18 4-075 NUCLEODUR C18 Gravity Macherey-Nagel 110 340 18 Syncronis C18 4-075 NUCLEODUR CN Macherey-Nagel 110 340 7 Hypersil GOLD CN 4-032

NUCLEODUR Pyramid Macherey-Nagel 110 340 14 Syncronis aQ 4-077 ™ Nucleoshell RP 18 Macherey-Nagel 90 130 7.5 Accucore C18 4-045 >>

Nucleoshell Phenyl-Hexyl Macherey-Nagel 90 130 4.5 Accucore Phenyl-Hexyl 4-049 Column Selection Nucleoshell PFP Macherey-Nagel 90 130 3 Accucore PFP 4-051 Nucleosil™ 100 C18 Macherey-Nagel 100 350 17 Syncronis C18 4-075 Nucleosil 100 C18 AB Macherey-Nagel 100 350 24 Syncronis C18 4-075

Nucleosil 100 C6H5 Macherey-Nagel 100 350 – Syncronis Phenyl 4-078 Nucleosil 100 C8 Macherey-Nagel 100 350 9 Syncronis C8 4-076 Nucleosil 100 CN Macherey-Nagel 100 350 – Hypersil GOLD CN 4-032

Nucleosil 100 N(CH3)2 Macherey-Nagel 100 350 – Hypersil SAX 4-109

Nucleosil 100 NH2 Macherey-Nagel 100 350 4 Syncronis Amino 4-079 Nucleosil 100 OH Macherey-Nagel 100 350 – BETASIL Diol Inquire Nucleosil 100 SA Macherey-Nagel 100 350 7 BioBasic SCX 4-141 Nucleosil 100 SB Macherey-Nagel 100 350 10 Hypersil GOLD SAX 4-035 Nucleosil 300 C18 Macherey-Nagel 300 100 7 BioBasic 18 4-140 Nucleosil 300 C4 Macherey-Nagel 300 100 – BioBasic 4 4-140

Nucleosil 300 C6H5 Macherey-Nagel 300 100 – BioBasic Phenyl Inquire Nucleosil 300 C8 Macherey-Nagel 300 100 – BioBasic 8 4-140 Nucleosil 300 CN Macherey-Nagel 300 100 – BioBasic CN Inquire Pinnacle™ C1 Restek 120 170 2 Hypersil SAS 4-107 Pinnacle C18 Restek 120 170 10 Hypersil GOLD 4-026 Pinnacle C4 Restek 120 170 4 Hypersil GOLD C4 4-028 Pinnacle CN Restek 120 170 5 Hypersil GOLD CN 4-032 Pinnacle DB C18 Restek 140 – 11 Hypersil GOLD 4-026 Pinnacle DB C18 1.9µm Restek 140 – 11 Hypersil GOLD (1.9µm) 4-026 Pinnacle DB C8 Restek 140 – 6 Hypersil GOLD C8 4-027 Pinnacle DB Cyano Restek 140 – 4 Hypersil GOLD CN 4-032 Pinnacle DB Phenyl Restek 140 – 5 Hypersil GOLD Phenyl 4-031 Pinnacle IBD Restek 120 170 – Hypersil GOLD 4-026

Pinnacle NH2 Restek 120 170 2 Hypersil GOLD Amino 4-033 Pinnacle Phenyl Restek 120 170 5 Hypersil GOLD Phenyl 4-031 Pinnacle SAX Restek 120 170 3 Hypersil GOLD SAX 4-035 Pinnacle Silica Restek 120 170 – Hypersil GOLD Silica 4-036 Pinnacle Ultra C18 Restek 100 – 20 Syncronis C18 4-075 Pinnacle Wide Pore C4 Restek 300 – 2 BioBasic 4 4-140 Poroshell™ 120 EC-C18 Agilent 120 120 8 Accucore C18 4-045 Poroshell 120 EC-C8 Agilent 120 120 5 Accucore C8 4-046 Poroshell 120 SB-C18 Agilent 120 120 7.5 Accucore C18 4-045 Poroshell 120 SB-Aq Agilent 120 130 – Accucore aQ 4-047

www.thermoscienti c.com/chromatography 4-013 4 -014 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories HPLC Column Selection by Manufacturer by Selection Column HPLC TSKgel TSKgel SuperSW3000 TSKgel Phenyl Super TSKgel ODS Super TSKgel Octyl Super TSKgel TSKgel ODS-80TM TSKgel ODS-120T TSKgel ODS-120A TSKgel Octyl-80TS Synergi C8 Symmetry SunShell C8 SunFire Symmetry Supelcosil LC-Si SunFire W(silica) Spherisorb Waters SCX Spherisorb Waters Waters Spherisorb SAX Phenyl Spherisorb Waters ODSB Spherisorb Waters ODS2 Spherisorb Waters ODS1 Spherisorb Waters Prodigy Poroshell 300 SB-C3 300 Poroshell SB-C8 300 Poroshell Poroshell Extend-C18 SB-C18 Poroshell 120Bonus-RP Poroshell Poroshell 120 Phenyl-Hexyl Supelcosil SunShell PFP SunShell C8 Supelcosil LC-NH Supelcosil LC-CN Supelcosil LC-8 Supelcosil LC-18DB Supelcosil LC-18 Waters Spherisorb NH Spherisorb Waters CN Spherisorb Waters C8 Spherisorb Waters C6 Spherisorb Waters Waters RP-18e STAR Purospher uope TR8 Merck STAR-8e Purospher Purospher Phenyl-3 Prodigy Prodigy ODS-3V Prodigy ODS-3 Prodigy ODS2 Primesep Phase ™ ™ ™ ™ ™ G2000SW (incl XL) (incl G2000SW C8 C18 Hydro-RP Spherisorb ™ ™ C18 ™ ™ ™ RP-18 C18 LC-1 2 ™ C1 2 Phenomenex Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Waters Waters Waters Waters Supelco Supelco Waters Waters Waters Waters Waters Waters Waters Waters SieLC Agilent Agilent Agilent Agilent Agilent Agilent ChromaNik ChromaNik ChromaNik Supelco Supelco Supelco Supelco Supelco Waters Waters Waters Waters Merck Merck Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Manufacturer 80 250 110 110 110 80 120 120 80 125 100 100 90 90 120 120 80 80 80 80 80 80 80 80 – 300 300 300 300 120 120 90 90 90 120 120 120 120 120 80 80 80 80 120 120 60 100 100 100 150 150 Pore Size (Å) Size Pore 475 – – – – 200 200 200 200 – 335 335 340 340 170 170 200 200 200 200 200 200 200 200 – 45 45 45 45 130 130 150 150 150 170 170 170 170 170 200 200 200 200 300 300 500 450 450 450 310 310 Area (m 2 continued /g) 19 – BioBasic SEC 300 SEC BioBasic Phenyl GOLD Hypersil C8 GOLD Hypersil – 3 8 5 15 120 SEC BioBasic 22 22 11 – 12 19 16 16 – Hypersil GOLD Amino GOLD Hypersil – – Phenyl GOLD Hypersil – – – 3 12 12 6 2 – 1.1 1.5 4 2.8 9.5 9 % Carbon 4.5 4.5 7 – Hypersil GOLD CN GOLD Hypersil C8 GOLD Hypersil – – 11 11 – 3 Hypersil GOLD CN GOLD Hypersil C8 GOLD Hypersil 3 6 5 2 – Hypersil GOLD C8 GOLD Hypersil – – – 10 16 16 18 13 Syncronis aQ Syncronis Hypersil GOLD Hypersil GOLD C18 Syncronis Hypersil GOLD C8 GOLD Hypersil Syncronis C8 Syncronis C18 Syncronis Syncronis C8 Syncronis C18 Syncronis Hypersil GOLD Silica GOLD Hypersil Silica GOLD Hypersil SCX BioBasic Hypersil SAX Hypersil GOLD Hypersil GOLD Hypersil GOLD APS-2 Hypersil Acclaim Columns Mixed-Mode Accucore 150-C4 Accucore 150-C4 Accucore 150-C18 Accucore 150-C18 Accucore Premium Polar Accucore Phenyl-Hexyl Accucore Scienti Recommended Thermo c Alternative Accucore PFP Accucore C8 Accucore C18 Accucore Hypersil GOLD Hypersil GOLD Hypersil SAS BETASIL C6 BETASIL Hypersil SAS Hypersil GOLD Hypersil GOLD Syncronis Phenyl Syncronis Hypersil GOLD C18 Syncronis C18 Syncronis C8 Syncronis 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Page 4 4 4 4 4 4 4 4 Inquire 4 4 4 4 4 4 4 4 4 4 4 -077 -026 -026 -075 -026 -027 -139 -031 -027 -139 -076 -075 -076 -075 -036 -036 -141 -109 -026 -026 -026 -108 -068 -033 -031 -145 -145 -144 -144 -048 -049 -051 -046 -045 -026 -026 -107 -032 -027 -107 -032 -027 -026 -027 -026 -078 -026 -075 -075 -076 LC Columns and Accessories

Phase Manufacturer Pore Size (Å) Area (m2/g) % Carbon Recommended Thermo Scienti c Alternative Page Vydac™ 201SP C18 Grace 90 – – Hypersil GOLD 4-026 Vydac 201SP Selectapore Grace 90 250 – Hypersil GOLD 4-026 90M C18 Vydac 201TP C18 Grace 300 – – BioBasic 18 4-140 Vydac 202TP C18 Grace 300 – – BioBasic 18 4-140 Vydac 208TP C8 Grace 300 – – BioBasic 8 4-140 Vydac 214TP Grace 300 – – BioBasic 4 4-140 Vydac 218TP Grace 300 – – BioBasic 18 4-140 Vydac 218WP Selectapore Grace 300 70 – BioBasic 18 4-140 300M C18 XBridge™ C18 Waters – – – Hypersil GOLD 4-026 XBridge C8 Waters – – – Hypersil GOLD C8 4-027 XBridge HILIC Waters 130 185 – Hypersil GOLD Silica 4-036 XBridge Phenyl Waters – – – Hypersil GOLD Phenyl 4-031 XTerra™ MS C18 Waters 125 180 16 Hypersil GOLD 4-026 XTerra MS C8 Waters 125 180 12 Hypersil GOLD C8 4-027 >>

YMCbasic™ YMC – – – Hypersil GOLD C8 4-027 Column Selection YMC-Pack™ C4 YMC 120 300 7 HyPURITY C4 4-109 YMC-Pack C8 YMC 120 300 10 Acclaim C8 4-062 YMC-Pack CN YMC 120 300 7 Hypersil GOLD CN 4-032 YMC-Pack Diol YMC 120 300 – BETASIL Diol Inquire

YMC-Pack NH2 YMC 120 – – Hypersil GOLD Amino 4-033 YMC-Pack ODS AQ YMC 120 300 16 Syncronis aQ 4-077 YMC-Pack ODS-A YMC 120 300 17 Syncronis C18 4-075 YMC-Pack ODS-A YMC 300 150 6 BioBasic 18 4-140 YMC-Pack Phenyl YMC 120 300 9 Syncronis Phenyl 4-078 YMC-Pack Pro C18 YMC 120 350 16 Syncronis C18 4-075 YMC-Pack Silica YMC 120 – – Syncronis Silica 4-080 YMC-Pack TMS (C1) YMC 120 300 4 BETASIL C1 Inquire Zorbax™ Eclipse XDB C18 Agilent 80 180 10 Hypersil GOLD 4-026 Zorbax Eclipse XDB C8 Agilent 80 180 8 Hypersil GOLD C8 4-027 Zorbax Eclipse XDB Phenyl Agilent 80 180 8 Hypersil GOLD Phenyl 4-031 Zorbax Eclipse Plus C18 Agilent 95 160 8 Hypersil GOLD 4-026 Zorbax Eclipse Plus C8 Agilent 95 160 6 Hypersil GOLD C8 4-027 Zorbax RRHT Eclipse Plus C18 Agilent 95 160 8 Hypersil GOLD (1.9µm) 4-026 Zorbax RRHT Eclipse Plus C8 Agilent 95 160 6 Hypersil GOLD C8 (1.9µm) 4-027 Zorbax RRHT Eclipse Agilent 80 180 10 Hypersil GOLD (1.9µm) 4-026 XDB-C18 Zorbax RRHT Eclipse Agilent 80 180 7.5 Hypersil GOLD C8 (1.9µm) 4-027 XDB-C8 Zorbax RRHT SB-CN Agilent 80 180 4 Hypersil GOLD CN (1.9µm) 4-032 Zorbax SB Aq Agilent 80 180 – Hypersil GOLD aQ 4-029 Zorbax SB C18 Agilent 80 180 10 Hypersil GOLD 4-026 Zorbax SB C18 Agilent 300 45 3 BioBasic 18 4-140 Zorbax SB C8 Agilent 80 180 6 Hypersil GOLD C8 4-027 Zorbax SB C8 Agilent 300 45 2 BioBasic 8 4-140 Zorbax SB CN Agilent 80 180 4 Hypersil GOLD CN 4-032 Zorbax SB Phenyl Agilent 80 180 6 Hypersil GOLD Phenyl 4-031

www.thermoscienti c.com/chromatography 4-015 4 -016 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Choosing aguardorfilterbasedonsamplemake-up should as possible,andthepackingmaterial The internaldiametersshouldmatchas closely choose onethatmatchesyouranalyticalcolumn. simple ruleofthumbinguardselectionisto the samplereachesanalyticalcolumn.The strongly adsorbedsamplecomponentsbefore injector andtheanalyticalcolumn,removing Guard columnsarepositionedbetweenthe Guard andFilterSelection during theanalysisofprocainamides. unaffected bytheadditionofaguardunit column’s chromatographicperformanceis chromatogram showndemonstrateshowthe a guard or filter unittotheHPLCsystem.The should notbeaffectedbyadding performance of afilterorpackedguards.Column use these unwantedsystemcomponentsisby most cost-effectiveandefficientwayoftrapping and interferencesfromthesamplematrix.The contamination bysampleandsolventdebris recommend thattheybeprotectedfrom analytical andpreparativecolumns,we your To extendthelifetimeandperformanceof • • • • • Extend columnlifetimeandimproveperformance Protection Column Thermo Scienti c LCColumns UHPLC FiltercartridgesandholdertoprotectHypersilGOLD1.9µmcolumns Drop-in designsforquickandeasyguard lter replacement Multiple formatsforoptimumperformanceandef ciency Economical extensionofcolumnlifetime Guards and lterstoprotectyouranalyticalcolumn Sample Particulates Interferences Particulates andInterferences be thesameparticlesizeandchemistryas to anylengthcolumn. increase inbackpressure,socanbefitted dispersion. TheUHPLCfilteraddsaminimal causes minimalefficiencylossthrough and shape performance without degrading peak dead volumedesignmaintainschromatographic particles, enhancingcolumnlifetime.Itslow and Syncronis1.7µmcolumnsagainst filter cartridgesprotectHypersilGOLD1.9µm Replaceable 0.2µmThermoScientificUHLPC replaceable filtersinare-useableholder. designed tobewhollydisposableorhave dissolved inthefluidpath.Theseunitsare interferences orcontaminantsthatare the fluidpath.Theydonotremovesample are designedtotrapparticulatematterfrom solvent inletfilterandthecolumninlet.They Pre-column filtersarepositionedbetweenthe should besimpleandfast. is used,thereplacementofpackedcartridges analytical column.Ifaguardcartridgesystem be thesameparticlesizeandchemistryas Filter Samplethrougha0.45µm lter Isolate interferencesoranalytesusingSPE and/or and/or of aguardcolumn Peak resolutionandshapeareunaffectedbytheaddition Without aguard Without to trapparticulates Use apre-column lter any remaininginterferences Use apre-columnguardtotrap Analytes: Procainamides Analytes: Analytes: Procainamides Analytes: With C18 guard C18 With Analysis Reproducible Successful, Drop-in Guard Cartridges LC Columns and Accessories Drop-in guard cartridges offer convenience, economy and effective protection for extending column lifetime

• The 10mm design offers maximum protection with minimal increase in retention • Fit Thermo Scienti c™ UNIGUARD™ direct connection and stand alone holders

Drop-in Guard Cartridges Ordering Guide Particle Size (µm) Length (mm) 1.0mm ID 2.1mm ID 3.0mm ID 4.0/4.6mm ID Quantity Hypersil GOLD 3 10 25003-011001 25003-012101 25003-013001 25003-014001 4 Pack 5 10 25005-011001 25005-012101 25005-013001 25005-014001 4 Pack Hypersil GOLD C8 3 10 25203-011001 25203-012101 25203-013001 25203-014001 4 Pack

5 10 25205-011001 25205-012101 25205-013001 25205-014001 4 Pack

Hypersil GOLD aQ >> 3 10 25303-011001 25303-012101 25303-013001 25303-014001 4 Pack Column Protection 5 10 25305-011001 25305-012101 25305-013001 25305-014001 4 Pack Hypersil GOLD PFP 3 10 25403-011001 25403-012101 25403-013001 25403-014001 4 Pack 5 10 25405-011001 25405-012101 25405-013001 25405-014001 4 Pack Hypersil GOLD CN 3 10 25803-011001 25803-012101 25803-013001 25803-014001 4 Pack 5 10 25805-011001 25805-012101 25805-013001 25805-014001 4 Pack Hypersil GOLD Phenyl 3 10 25903-011001 25903-012101 25903-013001 25903-014001 4 Pack 5 10 25905-011001 25905-012101 25905-013001 25905-014001 4 Pack Accucore XL C18 4 10 – 74104-012101 74104-013001 74104-014001 4 Pack Accucore XL C8 4 10 – 74204-012101 74204-013001 74204-014001 4 Pack Syncronis C18 5 10 – 97105-012101 97105-013001 97105-014001 4 Pack Syncronis C8 5 10 – 97205-012101 97205-013001 97205-014001 4 Pack Syncronis aQ 5 10 – 97305-012101 97305-013001 97305-014001 4 Pack Syncronis Phenyl 5 10 – 97905-012101 97905-013001 97905-014001 4 Pack Hypercarb 3 10 35003-011001 35003-012101 35003-013001 35003-014001 2 Pack 5 10 35005-011001 35005-012101 35005-013001 35005-014001 2 Pack This table provides a sample of the guard cartridges for the most popular Thermo Scientific HPLC stationary phases. Drop-in guard cartridges are available in other Thermo Scientific phases. For information on guard cartridges for other Thermo Scientific phases, please consult the appropriate section of the catalogue or contact Customer Services for more information.

www.thermoscienti c.com/chromatography 4-017 4 -018 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories to workwithhighspeed,efficiencyseparations. • • eliminating requirementforextrafittings Reusable, stainlesssteelguardcartridgeholdersthatmatchdirectly totheanalyticalcolumninlet– Holders Guard Direct-Connection Cartridge UNIGUARD Thermo Scientific Cartridges Guard Defender Other guardcolumndimensionareavailableinsomeAccucorephases.PleasecallyourlocalCustomerServiceformoreinformation. 1/16in femaleinletcanbeusedwithvariousstandard ttings With PEEK1/16inmaleoutletthat tsmostcolumns Waters ColumnsWaters Replacement Tip Standard Replacement Tip Replacement Standard Guard Cartridge Holder Guard Cartridge UNIGUARD /Defender Drop-In Description Guide Ordering Holders Cartridge Guard Direct-Connection UNIGUARD Phenyl-X Accucore Phenyl-Hexyl Accucore Premium Polar Accucore aQ Accucore C8 Accucore C18 Accucore RP-MS Accucore Description Accucore Defender Guard Cartridges (4/pk) Ordering Guide Ordering (4/pk) Cartridges Guard Defender Accucore ™ Defender 2.6 2.6 2.6 2.6 2.6 2.6 2.6 (µm) Size Particle ™ GuardCartridgeshavebeendesignedspecifically 10 10 10 10 10 10 10 (mm) Length 5-T80W 5-T850-WT 850-WT 850-WT 850-WT 850-RT 851-00 1.0mm ID 27926-012105 17926-012105 28026-012105 17326-012105 17226-012105 17126-012105 17626-012105 2.1mm ID 850-RT 852-00 2.1mm ID 2.1mm Accucore 150-Amide-HILIC Accucore 150-C4 Accucore 150-C18 Accucore Accucore Urea-HILIC HILIC Accucore C30 Accucore PFP Accucore Description 850-RT 852-00 .m D4.0/4.6mm ID 3.0mm ID 2.6 2.6 2.6 2.6 2.6 2.6 2.6 (µm) Size Particle 850-RT 850-00 10 10 10 10 10 10 10 (mm) Length 16726-012105 16526-012105 16126-012105 27726-012105 17526-012105 27826-012105 17426-012105 2.1mm ID 1 Each 1 Each 1 Each Quantity Acclaim Guards Cartridges LC Columns and Accessories Thermo Scientific™ Acclaim™ Guard Cartridges are available in the same packing and internal diameters as the Acclaim analytical column chemistries. A re-usable holder allows for easy cartridge replacement; extending the life of the analytical column.

Acclaim Guard Cartridges Ordering Guide

Acclaim Guard Cartridge Ordering Guide (2/pk) Particle Size Length ID 120 C18 120 C8 Polar Polar C30 Mixed-Mode Mixed-Mode Mixed-Mode (µm) (mm) (mm) Advantage Advantage II HILIC-1 WAX-1 WCX-1 5 10 2.1 069689 069688 069691 069692 075722 069694 069686 085455 3.0 071981 071979 071983 071985 075721 071913 071909 071911 4.6 069695 069696 069698 069699 075720 069706 069704 069705

Acclaim Guard Cartridge Ordering Guide (2/pk) continued >>

Description Particle Size Length 2.1mm ID 3.0mm ID 4.6mm ID Column Protection (µm) (mm) Phenyl-1 3 10 079934 071974 071973 HILIC-10 5 10 074263 074261 074262

Acclaim Specialty Column Guard Cartridge Ordering Guide (2/pk) Description Particle Size Length 2.1mm ID 3.0mm ID 4.6mm ID (µm) (mm) Acclaim Trinity P1 3 10 071391 071390 – Acclaim Trinity P2 3 10 085435 085436 – Acclaim Trinity Q1 5 10 079719 079720 – Acclaim Organic Acid 5 10 – 071987 069700 Acclaim Surfactant Plus 5 10 082773 078959 078960 Acclaim Surfactant 5 10 069693 071991 069701 Acclaim Explosives E1 5 10 – – 069702 Acclaim Explosives E2 5 10 – 071989 069703 Acclaim Carbamate 5 10 072930 072929 072928 Acclaim Carbonyl C18 5 10 079012 079013 079014

Acclaim Guard Holder Ordering Guide Format Cat. No. Acclaim Guard Cartridge Holder V-2 069580 Acclaim Guard Kit (Holder and coupler) V-2 069707 Guard to Analytical Column Coupler V-2 074188

www.thermoscienti c.com/chromatography 4-019 4 -020 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • without compromisingperformance Column protectionforHypersilGOLD1.9µmandSyncronis 1.7µm columns Filter UHPLC • • Quickly replacedforminimaldowntime Systems Filter HPLC Direct-Connection UNIFILTER Filter meshsize2μm ColumnSaver Precolumn Filters • • • • • One-piece filterprotectsHPLCsystems Filters Column Javelin Direct-Connection UHPLC Filter Ordering Guide Ordering Filter UHPLC Guide Ordering Systems Filter HPLC Direct-Connection UNIFILTER Guide Ordering Filters Precolumn ColumnSaver Guide Ordering Filter Column Direct-Connection Javelin Minimal ef ciencylossthrough dispersion Maintain peakshape Low volume ltercartridge design Holder attacheddirectlytotheinletofyouranalyticalsystemformaximumconvenience Replaceable 0.5µmdrop-in lterenhancescolumnlifetimeandimprovedperformance porosity 0.5µm 1/16in CPItipattachesdirectlytoHPLCcolumninletwithouttubingorwrenches Recommended foruseasdedicated ltersforacolumnratherthantheHPLCsystem Replace entiredisposable lterunitforeasychanges Direct-connection designformaximumef ciency 1.0mm ID Replacement Filter Cartridge, 0.2μm Cartridge, Filter Replacement ID 1.0mm 0.2μm Cartridge, Filter 2.1mm Replacement ID Holder Filter Connect Direct UHPLC Description End- Waters Tip, tting Replacement Standard CPI, Tip, Replacement 0.5µm Filter, Replacement 0.5µm Filter, Replacement Holder Connection Direct UNIFILTER Description 2 (µm) Size Mesh Filter Description Javelin Column Filter Filter Column Javelin 2.1mm ID 2.1mm 80 88700 88200 850-WT 850-RT 22017 4.0/4.6mm ID 22016 27002 2.1/3.0mm ID .m D4.0/4.6mm ID 3.0mm ID 850-WT 850-RT 22155 22150 27000 22185 22180 27006 No. Cat. 60140-412 No Cat. 88400 1 Each 1 Each 5 Pack 1 Each 1 Each Quantity 5 Pack 5 Pack 1 Each Quantity Pack 10 Quantity Quantity 4 Pack Notes LC Columns and Accessories

>> Column Protection

www.thermoscienti c.com/chromatography 4-021 4 -022 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories for yourseparationneeds. are achieved.Usethistableandtheinformationinsubsequentcataloguepages tofindthebestThermoScientificLCcolumns The tablebelowshowsourmajorcolumnfamilieswithdetailsofrelativehydrophobicity, keyattributesandhowthe for fastandanalyticalHPLC. The ThermoScientificLCcolumnportfoliooffersacomprehensive rangeofhighqualityoptions for FastandAnalyticalHPLC Thermo Scienti c Columns Hydrophobicity Very High Very High Medium/High Low Page 4 Columns Acclaim Page 4 Columns GOLD Hypersil Page 4 Columns Hypercarb Page 4 Syncronis Columns Page 4 Accucore Columns -060 -024 -084 -074 -042 Application Speci c & Speciality Traditional, Shape Peak & Selectivity Capabilities Separation Extended Reproducibility & Ruggedness Performance Ultimate Key Attribute • • • • • • • • • • Achieved through Achieved Enhanced Core core solid 4µm and 2.6 Polar retention effect effect retention Polar porous 100% 5µm 3 and double bonding, Dense 1.7, high 5µm 3and unique and Innovative ultra-pure 5µm 3and 2.2, of range Wide 12µm 1.9, 5and 3, Technology particles silica on graphite (PREG) graphite on particles carbon graphitic rigorous endcapping, silica area surface bonded phases particles silica conventional phases silica purity ultra-high Technology particles silica on graphite (PREG) graphite on particles carbon graphitic testing rigorous endcapping, particles silica area surface bonded phases particles silica conventional phases particles silica purity ultra-high 15,000 Impedance Plot

3µm

5µm LC Columns and Accessories Principles of Fast LC Accucore 2.6µm

Effect of Increasing Impedance particle size and type An alternative to small diameter particles Pt0 It is now well established that columns packed is the Core Enhanced Technology used in E = 2 N with smaller diameter particles generate higher Accucore HPLC columns. As shown in the Ε Impedance efficiencies over1,500 a wider range of flow rates impedance plot below, Accucore columns ΔP Backpressure 45,000generate higher efficiencies in shorter times 4,500 than larger particle columns – as shown in t Retention time of unretained peak the plot below. than columns packed with 5µm or 3µm particles 0 Ν Ef ciency Increasing Ef ciencyand does so at low backpressures. η Mobile phase viscosity

15 15,000 Impedance Plot Advantage of Small Particles 5µm

u opt 3µm 10 3µm

u opt 5µm H (µm)

5 1.9µm u opt Accucore 2.6µm Increasing Column Ef ciency N) Increasing Impedance Pt0 >> E = 2 N 1,500 Columns for Analytical LC 0 1 2 3 4 5 6 45,000 4,500 u (mm/s) Increasing Flow Rate Increasing Ef ciency

Speed and Resolution Peak Capacity Miniaturization The general chromatographic resolution As an alternative to speeding up analysis the The sensitivity formula also shows that peak equation shows that resolution is directly excellent resolution offered by high efficiency concentration can be increased through the proportional to efficiency. High efficiencies columns can also be used to improve complex use of shorter columns and more importantly, across a wider range of linear velocities mean separations through an increase in peak with narrower column internal diameters. that shorter columns and/or faster flow rates capacity – the number of peaks that can be When transferring a method to a different can be used to increase the speed of separated in a given gradient time. column geometry adjustments must be made separations without sacrificing resolution. t to the following parameters: 1 + g The resolution equation also shows that a nc = ( ) • Flow Rate wide range of different bonded phases, each W • Injection Volume offering a different selectivity, is a useful way n Peak capacity c • Gradient Pro le to improve resolution. Gradient time tg A convenient method transfer tool is available 1 (α − 1) k' W Peak width (10% height) at the Chromatography Resource Center RS = N 4 α 1 + k' (www.thermoscientific.com/crc). α Selectivity Sensitivity Optimization N Ef ciency According to the formula shown below, sensitivity In order to preserve high efficiency, and k’ Capacity factor is increased in high efficiency separations by therefore resolution and sensitivity, the HPLC increasing the concentration of the peak and system in use should be optimized to reduce Poor Resolution thus the detector signal to noise ratio. any potential causes of peak broadening. See page 4-231 for details of this optimization. N Vi cmax∝ 2 L dc (1 + k' )

cmax Concentration at peak apex N Ef ciency Vi Injection volume L Column length

dc Column internal diameter k’ Capacity factor

Good Resolution

www.thermoscienti c.com/chromatography 4-023 4 -024 -024 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories extensive qualitycontroltesting. using arobustmanufacturingprocedureand accredited laboratoriesunderstrictprotocols columns aremanufacturedinISO9001:2008 for improvedchromatography. HypersilGOLD state-of-the-art familyofcolumns,designed continue toextendthecapabilitiesofthis HPLC mediaandcolumns,wesuccessfully product developmentandmanufacturingof Built onmorethan35yearsofexperiencein them an ideal choice for new method development. robust andstableforthelifeofassay, making developed withHypersilGOLDcolumnswillbe allows theusertobeconfidentthatassays chromatography, columnaftercolumn.This are exceptionallyreproducibleforreliable and maximizeproductivity: array ofchemistriestooptimizeseparations Hypersil GOLDcolumnsareavailableinan and Productivity Improved Selectivity, Resolution Thermo Scientific • • • • • Outstanding peakshapeforyourseparations Thermo Scienti c HypersilGOLDHPLCColumns 1.9 to12µmparticles Variety ofchemistries Increased sensitivityandimprovedresolution Narrow peaksforoutstandingef ciency Excellent peaksymmetry ™ HypersilGOLD ™ columns • • • • • • • •

reversed phaseandnormalphase. three modes:weakanionexchange, excellent chromatographicpropertiesin Hypersil GOLDAminodemonstrates retention length, lowhydrophobicitycolumnforless Hypersil GOLDC4hasshortalkylchain reversed andnormalphaseseparations Hypersil GOLDCNcanbeusedforboth selectivity inreversephaseapplications Hypersil GOLDPFPcanofferalternative employing highlyaqueousmobilephases challenging reversephaseseparations Hypersil GOLDaQcanbeusedfor but withlessretention Hypersil GOLDC8offerssimilarselectivity using genericgradientswithC18selectivity Hypersil GOLDoffersoutstandingpeakshape aromatic andmoderatelypolarcompounds selectivity andisparticularlysuitablefor Hypersil GOLDPhenyloffersalternative defines allThermoScientificcolumns. same careandattentiontoqualitythat development. Eachphaseismadewiththe elution orderforflexibilityinmethod providing enhancedretentionorchangesin selectivities inthesamecolumnfamily, These chemistriesofferalternative • • • •

mobile phase exchange column,designedforaqueous silica-based quarternaryaminestronganion Hypersil GOLDSAXisahighlystable species andpolarmolecules separate proteins,peptides,otheranionic Hypersil GOLDAXcanbeusedto phase columns analytes thatareproblematicusingreversed Hypersil GOLDHILICcolumnsretainpolar compounds bynormalphase non-polar andmoderatelypolarorganic ef cienttoolinthechromatographyof Hypersil GOLDSilicaisapowerfuland 1. Catechin Sample: 280nm at UV Temperature: 25°C 1mL/min 15 Bin min 50% to 20 Detection: Rate: Flow Gradient: acid +0.1% Formic H2O A: Phase: Mobile x4.6mm 150 5µm, GOLD, Hypersil 6. Catechin Gallate Catechin 6. 5. Epicatechin Gallate 4. Gallocatechin Gallate 3. Epicatechin Gallate Epigallocatechin 2. acid +0.1% Formic MeOH B: 11 LC Columns and Accessories Hypersil GOLD Signal 4 6 Competitor 2 8 300 3 2 7 9 10 5 Competitor 1 1 Competitor 18 column 200 0510 15 20 25 11 100

7 9 10 4 3 8 5 6 2 1 Hypersil GOLD column The improved peak symmetry provides additional 0510 15 20 25 peak height to increase sensitivity of analysis of trace components.

Resolution of analytes is improved using a Hypersil GOLD column. Data courtesy of M. Euerby, AstraZeneca, Charnwood, UK.

Solutions for High Throughput Screening, Enhanced peak height can be particularly critical Capillary to Preparative Analysis when low concentrations of an analyte are present, Hypersil GOLD columns are available in particle for example in an impurity assay. The increase sizes and column designs to meet all separation in sensitivity gained with the Hypersil GOLD >> needs, including improved resolution, enhanced columns over competitor C18 columns is Columns for Analytical LC sensitivity and faster analyses. From 1.9µm illustrated above. to 12µm particles, Hypersil GOLD columns offer Enhanced Resolution Excellent reproducibility is illustrated with the chromatographic solutions with consistent percent carbon on the Hypersil GOLD media Robust assay development requires a clear separations and performance. Specialized definition of resolution expectations. Narrow hardware includes Thermo Scientific™ KAPPA™ symmetrical chromato graphic peaks ensure capillary columns, PicoFrit™ and IntegraFrit that optimum resolution is achieved. Obtaining nanobore columns, Thermo Scientific™ Javelin™ narrow peak widths is especially challenging HTS direct-connection columns and for basic pharmaceutical compounds. The figure Thermo Scientific™ DASH™ HTS columns, above shows how Hypersil GOLD columns provide designed for high throughput screening. excellent resolution between critical pairs, aiding Improved Sensitivity in separation of closely related species. Good peak shape means greater sensitivity. pH Stability When peaks exhibit tailing, peak height is Hypersil GOLD columns are well suited to reduced causing the sensitivity of the analysis Stability of Hypersil GOLD columns at low pH. extended pH applications. Hypersil GOLD to be compromised. The more symmetrical the No loss of retention after 28L of mobile phase in columns have been shown to produce robust 19.5 days of analysis. chromatographic peaks, the more confidence assays at high pH. At low pH, excellent column you derive from your data. Using Hypersil GOLD stability and reproducibility are illustrated. columns, peak height is enhanced and peak integration calculations are optimized.

3 1. Uracil 2. Acetaminophen 3. N-Acetyl procainamide 4. n-Propionyl procainamide 2 H250-001 Dimensions: 5µm, 150 x 4.6mm

4 Part Number: 25005-154630 Mobile Phase: A: 0.1% ammonia pH 10.6 B: MeOH + 0.1% ammonia Gradient: 5 – 100% B in 15 min 5 6 Flow: 1.0mL/min 5. Oxazepam 6. Diazepam 7. Diisopropylphthalate 8. Di-n-propylphthalate Injection: 10µL 1 Detection: UV at 254nm 7 Temperature: 30°C 8

High pH stability assay (pH 10.6) of Hypersil GOLD columns

www.thermoscienti c.com/chromatography 4-025 4 -026 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • shape andresolutionforHPLCLC/MS Endcapped, ultrapure,silica-basedcolumnswithexceptionalpeak GOLD Hypersil Con denceintheaccuracyandqualityofanalyticaldata Excellent resolution,ef ciencyandsensitivity Signi cantreductioninpeaktailingwhileretainingC18selectivity

Relative Abundance 100 10 20 30 40 50 60 70 80 90 0 0.0 RT: 0.00-1.00 Seven β-blockersin1minute H2O /MeCN+0.1%foricgrad15to100%in1min;0.5ml/min;30C;1.9umGold20x2.1mm550C;3.5kV;20V Blockers24 MS Base Peak 1.02E5 NL: 0.1 0.2 SM: 7G 1 0.3 2 3 0.4 Minutes 0.5 4 5 6 H250-101 7 1.00.90.80.70.6 ytm Thermo Scienti c 0.5ml/min System: 1min Bin 100% 15 to Detection: +ESI Temperature: 30°C rate: Flow Gradient: oiepae A–H phase: Mobile 25002-022130 Number: Part x2.1mm 20 GOLD Hypersil 1.9µm Column: 1. Atenolol Analytes: B – MeCN+0.1%formic acid B –MeCN+0.1%formic 7. Propanolol 6. Oxprenolol 5. Metoprolol 4. Timolol 3. Pindolol 2. Nadolol 2 O+0.1%formic acid O+0.1%formic Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low ™ Surveyor ™ MSQ 1.9μm, 3μm,5μm,12μm 10 1 to11 175 USP High 300 L1 25 14 LC Columns and Accessories Hypersil GOLD C8 Hydrophobicity

Recommended for analytes with medium hydrophobicity or when a Low High less hydrophobic phase is required to obtain optimum retention pH Range

0 2 to 9 14 Pore Size • Similar selectivity to C18 columns but with reduced retention • Lower hydrophobicity, allowing compounds to elute quicker 0 175 300 Carbon Load (%) • Faster separations • Excellent peak shape 0 8 25 • High ef ciency Particle Size 1.9μm, 3μm, 5μm USP L7 • Outstanding sensitivity

β-carotene

600 1 Column: Hypersil GOLD C8, 5µm, 150 x 4.6mm

Part Number: 25205-154630

Mobile phase: MeOH >> 500 Flow rate: 1.5mL/min

Temperature: 25°C Columns for Analytical LC 400 Detection: UV @ 450 nm Analytes: 1. Lutein 2. Lycopene 300 3. β-Carotene mAU 200

2 3 100

0

0 1 2 3 4 5 6 7 Minutes H252-009

www.thermoscienti c.com/chromatography 4-027 4 -028 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • • analytes hydrophobic Lower hydrophobicitythanC18orC8recommendedforvery C4 GOLD Hypersil 120 160 200 mAU sensitivity Outstanding High ef ciency Excellent peakshape separations Faster hydrophobicity Lower 40 80 0 0.0 Fatty acids 0.2 0.4 0.6 1 2 0.8 Minutes 1.0 1.2 1.4 1.6 1.8 H255-002 2.0 ape 1. acid Linolenic nm 200 1µL Sample: volume: 0.55mL/min Injection Detection: Temperature: 30°C rate: Flow oiepae H phase: Mobile 25502-102130 Number: Part x2.1mm 100 1.9µm, C4 GOLD Hypersil Column: 2. acid Linoleic 2 O / MeCN (20:80) O /MeCN Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 Low 0 5 1.9μm, 3μm,5μm 2 to8 175 USP L26 High 300 25 14 LC Columns and Accessories Hypersil GOLD aQ Hydrophobicity

Hypersil GOLD aQ polar endcapped C18 columns provide a controlled Low High interaction mechanism by which polar analytes can be retained pH Range and resolved 0 2 to 9 14 Pore Size

• Polar endcapped C18 phase for alternative selectivity 0 175 300 Carbon Load (%) • Retention and resolution of polar analytes

• Excellent peak shape 0 12 25 • Stable in 100% aqueous mobile phases Particle Size 1.9μm, 3μm, 5μm USP L1

Water soluble vitamins

1200 3 Column: Hypersil GOLD aQ, 5µm, 150 x 4.6mm 2 Part Number: 25305-154630 8 6 Mobile phase: A: 50 mM KH2PO4, pH 3.5

1000 B: MeOH

Gradient: 0 – 100% B in 15 min >> 800 Flow Rate: 1mL/min

1 Detection: UV @ 205nm Columns for Analytical LC Sample: 1. Vitamin B1 (thiamine) 600 2. Vitamin B6 (pyridoxine) 3. Vitamin B3 (nicotinamide) mAU 4. Vitamin B5 (pantothenic acid) 400 5. Folic Acid 6. Vitamin B12 (cyanocobalamin) 4 200 7 7. Vitamin H (biotin) 5 8. Vitamin B2 (ribo avin)

0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Minutes H253-003

www.thermoscienti c.com/chromatography 4-029 4 -030 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • • significant changesinsolute-stationaryphaseinteraction Introduction ofafluorinegroupintothestationaryphasecauses PFP GOLD Hypersil Excellent peakshapeandsensitivity Selectivity fornon-halogenatedpolarcompounds Extra retentionforhalogenatedspecies Alternative selectivitytoC18 The  uorineatomsaroundthephenylringenhancepi-piinteractions

uAU with aromaticmolecules 10000 20000 30000 40000 50000 60000 70000 0 0.0 Polyphenols PPP3 HGOLDPF UV Channel A 6.87E4 NL: 0.5 1 1.0 2 1.5 Minutes 3 2.0 2.5 3.0 4 3.5 H254-012 ape 1. Pyrogallol Cell) Flow (2µL @280nm UV Sample: 0.5µL 0.5mL/min Injection Volume: Detection: Temperature: Acid 25°C 0.1% Acetic rate: Flow phase: Mobile 25402-052130 Number: Part x2.1mm 50 1.9µm, PFP GOLD Hypersil Column: 4. Phenol 3. Resorcinol 2. Hydroquinone Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 1.9μm, 3μm,5μm 8 2 to8 175 USP High L43 300 25 14 LC Columns and Accessories Hypersil GOLD Phenyl Hydrophobicity

Contains a C4 linker which allows for superior alignment of the Low High phenyl ring with aromatic molecules pH Range

0 2 to 8 14 Pore Size • Enhanced pi-pi interactions with aromatics • Moderate hydrophobicity 0 175 300 Carbon Load (%) • Outstanding peak shape and sensitivity

0 8 25 Particle Size 1.9μm, 3μm, 5μm USP L11 Antidepressants

100 Column: Hypersil GOLD Phenyl 1.9µm, 50 x 2.1mm 1 80 3 Part Number: 25902-052130 Mobile phase: A – 0.1% Formic acid 5 B – 0.1% Formic acid in MeCN 60

Gradient: 10 – 60% B in 3.4mins, 60 - 90% B in 0.24 min >> 6 Flow rate: 0.5mL/min

mAU 2

40 Temperature: 60°C Columns for Analytical LC Injection Volume: 0.7µL 7 8 Detection: UV @ 225 and 254 nm 20 4 Analytes: 1. Uracil imp 2. Acetaminophen 3. p-Hydroxybenzoic acid 0 4. o-Hydroxybenzoic acid 0.0 1.0 2.0 3.0 4.0 5. Oxazepam Minutes H259-006 6. Diazepam 7. Di-isopropyl phthalate 8. Di-n-propyl phthalate

www.thermoscienti c.com/chromatography 4-031 4 -032 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • reversed phaseseparations Hypersil GOLDCNcolumnscanbeusedforbothnormalphaseand CN GOLD Hypersil Less retentionforfasteranalysis senstivity Outstanding Excellent peakshape Provide alternativeselectivitywithlowerhydrophobicity 70 50 55 mVolts60 65 0.0 Organic acids 2.5 5.0 1 2 7.5 3 Minutes 10.0 4 5 12.5 15.0 H258-002 17.5 1. 4-Fluorobenzoic @230nm UV Sample: 1.5mL/min 25°C B 5% A: 95% Detection: Temperature: Rate: Flow Isocratic: Mobile Phase: A: 25 mM KH mM 25 A: Phase: Mobile 25805-154630 Number: Part x4.6mm 150 5µm, CN, Column: GOLD Hypersil B: MeOH 5. 2,5-Dimethylbenzoic Acid 2,5-Dimethylbenzoic 5. 4. 2,4,6-Trimethylbenzoic Acid Acid p-Toluic 3. Acid o-Toluic 2. 2 PO 4 pH2 Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 4 1.9μm, 3μm,5μm 2 to8 175 USP High L10 300 25 14 LC Columns and Accessories Hypersil GOLD Amino Hydrophobicity

A high performance aminopropyl phase that gives excellent Low High chromatographic properties in three modes: weak anion exchange, pH Range reversed phase and normal phase 0 2 to 8 14 Pore Size

• Retains anions and organic acids in weak anion exchange 0 175 300 Carbon Load (%) • Excellent for carbohydrate analysis in reversed phase

• Alternative selectivity to silica columns in normal phase chromatography 0 2 25 • Outstanding peak shape and sensitivity Particle Size 1.9μm, 3μm, 5μm USP L8

Sugars

14 Column: Hypersil GOLD Amino, 5µm, 150 x 4.6mm 1 Part Number: 25705-154630

12 >> Mobile Phase: MeCN/Water (80:20) Flow Rate: 1.2mL/min

10 Columns for Analytical LC Temperature: 35°C Detection: RI 8 Injection volume: 20 µL 3 Sample: 1. Fructose

uRIU 6 2 2. Glucose 4 4 3. Sucrose 4. Maltose 2 5. Lactose 5 0

0 2 4 6 8 10 12 14 16 18 20 Minutes H257-001

www.thermoscienti c.com/chromatography 4-033 4 -034 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • silica deactivated A novelpolymericamineligandbondedtohighlypurebase AX GOLD Hypersil 100 Outstanding peakshapeandselectivity Higher ef ciencythanpolymerbasedionexchangecolumns Suitable forHILICretentionandseparationofhighlypolarmolecules Weak anionexchangephaseformultiplechargedspecies

20 40 mAU 60 80 0 0 Vitamin C 5 10 Minutes 15 20 H261-002 25 Detection: UV @240nm UV 50µL Injection volume: 0.5mL/min 30°C Detection: Temperature: Rate: Flow 6.8/ pH Acetate Ammonium mM 100 Phase: Mobile 26105-104630 Number: Part x4.6mm 100 5µm, AX, GOLD Hypersil Column: MeCN (30:70) Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 6 1.9μm, 3μm,5μm 2 to8 175 High 300 25 14 LC Columns and Accessories Hypersil GOLD SAX Hydrophobicity

A highly stable quaternary amine strong anion exchange column for Low High aqueous and low pH mobile phases pH Range

0 2 to 8 14 Pore Size • High stability to aqueous and low pH mobile phases • Ideally suited to the analysis of smaller organic molecules including 0 175 300 nucleotides and organic acids Carbon Load (%) • Outstanding peak shape and sensitivity 0 2.5 25 Particle Size 1.9μm, 3μm, 5μm USP L14

Monophosphates

200 Column: Hypersil GOLD SAX, 5µm, 150 x 4.6mm Part Number: 26105-154630 175

Mobile Phase: Aqueous KH2PO4 (50 mM, pH 3) Flow Rate: 1.0mL min-1 150 >> Temperature: 40°C 125

Detection: UV @ 254nm Columns for Analytical LC Injection volume: 10µL 100

75 mAU 50

25

0

0 1 2 3 4 5 6 7 8 Minutes H263-001

www.thermoscienti c.com/chromatography 4-035 4 -036 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • backbone oftheHypersilGOLDrangecolumns Unbonded, highlypurebasedeactivatedsilicamediathatisthe Silica GOLD Hypersil

mVolts Outstanding peakshapeandsenstivity Highly purebasedeactivatedsilicamedia 10 20 30 40 50 60 70 80 90 0 0 Steroids 1 2 3 4 5 6 Minutes 7 8 9 10 11 12 13 H251-002 14 15 1. Progesterone @254nm UV Sample: 5µL Injection volume: min-1 1.5mL 30°C Detection: Temperature: n-C6H14/EtOH (v/v) 19:1 Rate: Flow Phase: Mobile 25105-154630 Number: Part x4.6mm 150 5µm, Silica, GOLD Hypersil Column: www.thermoscienti www.thermoscienti c.com/hypersilgold c.com/hypersilgold column range.Visit: downloads fortheHypersilGOLDHPLC the latestnews,applicationsand The HypersilGOLDwebpagecontains 7. Hydrocortisone 6. Corticosterone 5. 11-a-Hydroxyprogesterone 4. Cortisone 3. 17-a-Hydroxyprogesterone 2. 21-Hydroxyprogesterone-21-acetate Particle Size Pore Size pH Range Hydrophobicity 0 Low 0 1.9μm, 3μm,5μm 2 to8 175 USP High 300 L3 14 LC Columns and Accessories Hypersil GOLD HILIC Hydrophobicity

Hypersil GOLD HILIC retains and separates polar analytes that are Low High problematic using reversed phase columns pH Range

0 2 to 8 14 Pore Size • Alternative selectivity to C18 • Improved sensitivity for MS detection 0 175 300 Carbon Load (%) • Alternative to ion-pair or derivatisation

• Outstanding peak shape and selectivity 0 6 25 Particle Size 1.9μm, 3μm, 5μm USP L14

Urea

100 Column: Hypersil GOLD HILIC, 5µm, 150 x 4.6mm Part Number: 26505-154630

80 Mobile Phase: H2O/MeCN (10:90) + 0.1% formic adic Flow Rate: 0.6mL/min

60 Temperature: 30°C >> Detection: +ESI

Injection volume: 1µL (made up in mobile phase) Columns for Analytical LC 40 Sample: 1. Urea

20 Relative Abundance

0 0 1 2 3 4 5 6 7 8 9 10 Minutes H265-005

www.thermoscienti c.com/chromatography 4-037 4 -038 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Hypersil GOLD Ordering Guide Ordering GOLD Hypersil 5 3 1.9 (µm) Size Particle HPLC Column HPLC Column HPLC Drop-in Guard (4/pk) Drop-in Guard (4/pk) UHPLC Column Format 250 150 100 50 150 100 50 200 150 100 50 30 30 30 10 10 20 (mm) ID (mm) Length 2.1 2.1 2.1 2.1 1.0 1.0 2.1 2.1 2.1 1.0 1.0 1.0 2.1 2.1 2.1 1.0 2.1 4.6 4.0 3.0 4.6 4.0 3.0 4.6 3.0 4.6 3.0 4.6 3.0 4.0/4.6 3.0 4.6 4.0 3.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 3.0 4.6 3.0 4.0/4.6 3.0 2.1 3.0 2.1 4.6 3.0 2.1 2.1 50-510225223 50-51025405-252130 25305-252130 25205-252130 25005-252130 25405-152130 25305-152130 25205-152130 25005-152130 25405-102130 25305-102130 25205-102130 25005-102130 25405-052130 25305-052130 25205-052130 25005-052130 25203-151030 25003-151030 25003-101030 25003-052130 25002-202130 25402-101030 25002-152130 25302-101030 25202-101030 25002-101030 25402-051030 25302-051030 25202-051030 25002-051030 25002-031030 25005-032130 25003-032130 50-111225020 50-11125405-012101 25305-012101 25205-012101 25005-012101 25403-011001 25303-011001 25203-011001 25003-011001 25402-022130 25302-022130 25202-022130 25002-022130 GOLD 50-560225243 50-56025405-254630 25305-254630 25405-254030 25205-254630 25305-254030 25405-253030 25005-254630 25205-254030 25305-253030 25005-254030 25205-253030 25405-154630 25005-253030 25305-154630 25405-154030 25205-154630 25305-154030 25005-154630 25205-154030 25005-154030 25205-153030 25405-104630 25005-153030 25305-104630 25205-104630 25005-104630 25405-054630 25005-103030 25305-054630 25405-053030 25205-054630 25305-053030 25005-054630 25205-053030 25005-053030 25005-034630 25405-014001 25005-033030 25305-014001 25405-013001 25205-014001 25305-013001 25005-014001 25205-013001 25005-013001 25003-154630 25003-154030 25003-153030 25203-152130 25003-152130 25003-104630 25203-104030 25003-104030 25203-103030 25003-103030 25403-054630 25003-102130 25303-054630 25203-054630 25003-054630 25003-054030 25003-053030 25403-014001 25003-034630 25203-033030 25403-013001 25303-014001 25003-033030 25403-012101 25303-013001 25203-014001 25303-012101 25203-013001 25003-014001 25203-012101 25003-013001 25003-012101 25402-102130 25302-102130 25002-103030 25202-102130 25402-054630 25002-102130 25302-054630 25202-054630 25402-052130 25002-054630 25202-053030 25302-052130 25002-053030 25202-052130 25402-032130 25002-052130 25302-032130 25202-032130 25002-032130 25203-101030 25203-052130 25202-202130 25202-152130 – – 25203-032130 C8 25205-103030 – – 25203-154630 25203-154030 25203-153030 25203-104630 25203-102130 25203-054030 25203-053030 25203-034630 25202-103030 25303-151030 25403-101030 25303-101030 25403-052130 25303-052130 25402-202130 25302-202130 25302-152130 – – 25403-032130 25303-032130 aQ 25305-153030 25305-103030 – – 25303-154630 25303-154030 25403-153030 25303-153030 25303-152130 25303-104630 25303-104030 25303-103030 25303-102130 25403-054030 25303-054030 25303-053030 25303-034630 25303-033030 25402-103030 25302-103030 25302-053030 25403-151030 25402-152130 – – PFP 25405-153030 25405-103030 – – 25403-154630 25403-154030 25403-152130 25403-104630 25403-104030 25403-103030 25403-102130 25403-053030 25403-034630 25403-033030 25402-053030 50-560295243 50-560275243 60-560235243 25105-254630 26305-254630 25105-254030 26105-254630 26305-254030 25705-254630 26105-254030 25105-252130 25805-254630 25705-254030 26105-253030 26305-252130 25905-254630 25805-254030 25705-253030 26105-252130 25505-254630 25905-254030 25805-253030 25705-252130 25505-254030 25905-253030 25805-252130 25705-154630 25505-253030 25905-252130 25805-154630 25505-252130 25905-154630 25505-154630 – 25805-152130 25705-104630 – 25905-152130 25805-104630 25505-152130 25905-104630 25505-104630 25905-103030 25805-102130 25705-054630 25505-103030 25905-102130 25805-054630 25505-102130 25905-054630 25705-052130 25505-054630 25805-052130 – 25905-052130 25505-052130 – – 25805-014001 – 25905-014001 25805-013001 25505-014001 25905-013001 25805-012101 25505-013001 25905-012101 25505-012101 25903-154630 25503-154630 – 25903-153030 25503-153030 25903-152130 25503-152130 25903-151030 26103-103030 25503-151030 25903-104630 25703-103030 25503-104630 25803-103030 – 25903-103030 25503-103030 25903-102130 25503-102130 – – – – 25903-052130 25503-052130 25903-034630 25503-034630 – 25903-032130 25503-032130 25903-014001 25503-014001 25903-013001 25503-013001 25903-012101 25503-012101 25903-011001 25503-011001 25902-202130 25502-202130 25902-152130 25502-152130 – 25902-102130 25502-102130 – – 25802-052130 – 25902-052130 25502-052130 – – – – C4 – – – – – – – – – – – – – – – – – – – – – Phenyl – – 25805-103030 – – – – 25103-153030 26303-153030 25803-154630 26103-153030 – 25703-153030 25803-153030 25803-152130 25703-151030 25803-151030 25803-104630 – 25803-102130 – – – 26503-014001 – 26503-013001 25103-014001 25803-052130 25103-013001 26303-014001 25803-034630 26303-013001 26103-014001 – 26103-013001 25703-014001 25803-032130 25703-013001 25803-014001 25703-012101 25803-013001 25703-011001 25803-012101 25803-011001 25802-202130 25802-152130 – 25802-102130 – – – – – – – CN 25705-152130 25705-102130 26305-014001 26305-013001 26105-014001 26105-013001 25705-014001 25705-013001 25705-012101 25702-052130 – – 25705-103030 – – – – 26103-154630 25703-154630 – 25703-152130 25703-104630 – 25703-102130 – – – – 25703-052130 25703-034630 – 25703-032130 25702-202130 25702-152130 – 25702-102130 – – – – – – – Amino 26105-152130 26105-102130 26105-012101 26102-052130 26105-154630 – – 26105-104630 26105-103030 26305-054630 26105-054630 – 26305-052130 26105-052130 – – – – 26103-152130 26103-151030 26103-104630 – 26103-102130 – – – – 26103-052130 26303-034630 26103-034630 – 26103-032130 26103-012101 26103-011001 26302-202130 26102-202130 26102-152130 – 26102-102130 – – – – – – – AX 26305-253030 26305-152130 26305-102130 26305-012101 26303-103030 26302-052130 26305-154630 – – 26305-104630 26305-103030 – – – – 26303-154630 – 26303-152130 26303-151030 26303-104630 – 26303-102130 – – – – 26303-052130 – 26303-032130 26303-012101 26303-011001 26302-152130 – 26302-102130 – – – – – – – SAX 25105-014001 25105-013001 25105-253030 25105-152130 25105-102130 25105-012101 26503-103030 25103-103030 25102-052130 25105-154630 – – 25105-104630 25105-103030 25105-054630 – 25105-052130 – – – 25103-154630 – 25103-152130 25103-151030 25103-104630 – 25103-102130 – – – – 25103-052130 25103-034630 – 25103-032130 25103-012101 25103-011001 25102-202130 25102-152130 – 25102-102130 – – – – – – – Silica 26505-014001 26505-013001 26505-254630 26505-254030 26505-253030 26505-252130 26505-152130 26505-102130 26505-012101 26503-153030 26502-052130 26505-154630 – – 26505-104630 26505-103030 26505-054630 – 26505-052130 – – – 26503-154630 – 26503-152130 26503-151030 26503-104630 – 26503-102130 – – – – 26503-052130 26503-034630 – 26503-032130 26503-012101 26503-011001 26502-202130 26502-152130 – 26502-102130 – – – – – – – HILIC LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) GOLD C8 aQ PFP C4 Phenyl CN Amino AX SAX Silica HILIC 1.9 UHPLC Column 20 2.1 25002-022130 25202-022130 25302-022130 25402-022130 – – – – – – – – 30 1.0 25002-031030 – – – – – – – – – – – 2.1 25002-032130 25202-032130 25302-032130 25402-032130 – – – – – – – – 50 1.0 25002-051030 25202-051030 25302-051030 25402-051030 – – – – – – – – 2.1 25002-052130 25202-052130 25302-052130 25402-052130 25502-052130 25902-052130 25802-052130 25702-052130 26102-052130 26302-052130 25102-052130 26502-052130 3.0 25002-053030 25202-053030 25302-053030 25402-053030 – – – – – – – – 4.6 25002-054630 25202-054630 25302-054630 25402-054630 – – – – – – – – 100 1.0 25002-101030 25202-101030 25302-101030 25402-101030 – – – – – – – – 2.1 25002-102130 25202-102130 25302-102130 25402-102130 25502-102130 25902-102130 25802-102130 25702-102130 26102-102130 26302-102130 25102-102130 26502-102130 3.0 25002-103030 25202-103030 25302-103030 25402-103030 – – – – – – – – 150 2.1 25002-152130 25202-152130 25302-152130 25402-152130 25502-152130 25902-152130 25802-152130 25702-152130 26102-152130 26302-152130 25102-152130 26502-152130 200 2.1 25002-202130 25202-202130 25302-202130 25402-202130 25502-202130 25902-202130 25802-202130 25702-202130 26102-202130 26302-202130 25102-202130 26502-202130 3 Drop-in Guard (4/pk) 10 1.0 25003-011001 25203-011001 25303-011001 25403-011001 25503-011001 25903-011001 25803-011001 25703-011001 26103-011001 26303-011001 25103-011001 26503-011001

2.1 25003-012101 25203-012101 25303-012101 25403-012101 25503-012101 25903-012101 25803-012101 25703-012101 26103-012101 26303-012101 25103-012101 26503-012101

3.0 25003-013001 25203-013001 25303-013001 25403-013001 25503-013001 25903-013001 25803-013001 25703-013001 26103-013001 26303-013001 25103-013001 26503-013001 >> 4.0/4.6 25003-014001 25203-014001 25303-014001 25403-014001 25503-014001 25903-014001 25803-014001 25703-014001 26103-014001 26303-014001 25103-014001 26503-014001 Columns for Analytical LC HPLC Column 30 2.1 25003-032130 25203-032130 25303-032130 25403-032130 25503-032130 25903-032130 25803-032130 25703-032130 26103-032130 26303-032130 25103-032130 26503-032130 3.0 25003-033030 25203-033030 25303-033030 25403-033030 – – – – – – – – 4.6 25003-034630 25203-034630 25303-034630 25403-034630 25503-034630 25903-034630 25803-034630 25703-034630 26103-034630 26303-034630 25103-034630 26503-034630 50 2.1 25003-052130 25203-052130 25303-052130 25403-052130 25503-052130 25903-052130 25803-052130 25703-052130 26103-052130 26303-052130 25103-052130 26503-052130 3.0 25003-053030 25203-053030 25303-053030 25403-053030 – – – – – – – – 4.0 25003-054030 25203-054030 25303-054030 25403-054030 – – – – – – – – 4.6 25003-054630 25203-054630 25303-054630 25403-054630 – – – – – – – – 100 1.0 25003-101030 25203-101030 25303-101030 25403-101030 – – – – – – – – 2.1 25003-102130 25203-102130 25303-102130 25403-102130 25503-102130 25903-102130 25803-102130 25703-102130 26103-102130 26303-102130 25103-102130 26503-102130 3.0 25003-103030 25203-103030 25303-103030 25403-103030 25503-103030 25903-103030 25803-103030 25703-103030 26103-103030 26303-103030 25103-103030 26503-103030 4.0 25003-104030 25203-104030 25303-104030 25403-104030 – – – – – – – – 4.6 25003-104630 25203-104630 25303-104630 25403-104630 25503-104630 25903-104630 25803-104630 25703-104630 26103-104630 26303-104630 25103-104630 26503-104630 150 1.0 25003-151030 25203-151030 25303-151030 25403-151030 25503-151030 25903-151030 25803-151030 25703-151030 26103-151030 26303-151030 25103-151030 26503-151030 2.1 25003-152130 25203-152130 25303-152130 25403-152130 25503-152130 25903-152130 25803-152130 25703-152130 26103-152130 26303-152130 25103-152130 26503-152130 3.0 25003-153030 25203-153030 25303-153030 25403-153030 25503-153030 25903-153030 25803-153030 25703-153030 26103-153030 26303-153030 25103-153030 26503-153030 4.0 25003-154030 25203-154030 25303-154030 25403-154030 – – – – – – – – 4.6 25003-154630 25203-154630 25303-154630 25403-154630 25503-154630 25903-154630 25803-154630 25703-154630 26103-154630 26303-154630 25103-154630 26503-154630 5 Drop-in Guard (4/pk) 10 2.1 25005-012101 25205-012101 25305-012101 25405-012101 25505-012101 25905-012101 25805-012101 25705-012101 26105-012101 26305-012101 25105-012101 26505-012101 3.0 25005-013001 25205-013001 25305-013001 25405-013001 25505-013001 25905-013001 25805-013001 25705-013001 26105-013001 26305-013001 25105-013001 26505-013001 4.0/4.6 25005-014001 25205-014001 25305-014001 25405-014001 25505-014001 25905-014001 25805-014001 25705-014001 26105-014001 26305-014001 25105-014001 26505-014001 HPLC Column 30 2.1 25005-032130 – – – – – – – – – – – 3.0 25005-033030 – – – – – – – – – – – 4.6 25005-034630 – – – – – – – – – – – 50 2.1 25005-052130 25205-052130 25305-052130 25405-052130 25505-052130 25905-052130 25805-052130 25705-052130 26105-052130 26305-052130 25105-052130 26505-052130 3.0 25005-053030 25205-053030 25305-053030 25405-053030 – – – – – – – – 4.6 25005-054630 25205-054630 25305-054630 25405-054630 25505-054630 25905-054630 25805-054630 25705-054630 26105-054630 26305-054630 25105-054630 26505-054630 100 2.1 25005-102130 25205-102130 25305-102130 25405-102130 25505-102130 25905-102130 25805-102130 25705-102130 26105-102130 26305-102130 25105-102130 26505-102130 3.0 25005-103030 25205-103030 25305-103030 25405-103030 25505-103030 25905-103030 25805-103030 25705-103030 26105-103030 26305-103030 25105-103030 26505-103030 4.6 25005-104630 25205-104630 25305-104630 25405-104630 25505-104630 25905-104630 25805-104630 25705-104630 26105-104630 26305-104630 25105-104630 26505-104630 150 2.1 25005-152130 25205-152130 25305-152130 25405-152130 25505-152130 25905-152130 25805-152130 25705-152130 26105-152130 26305-152130 25105-152130 26505-152130 3.0 25005-153030 25205-153030 25305-153030 25405-153030 – – – – – – – – 4.0 25005-154030 25205-154030 25305-154030 25405-154030 – – – – – – – – 4.6 25005-154630 25205-154630 25305-154630 25405-154630 25505-154630 25905-154630 25805-154630 25705-154630 26105-154630 26305-154630 25105-154630 26505-154630 250 2.1 25005-252130 25205-252130 25305-252130 25405-252130 25505-252130 25905-252130 25805-252130 25705-252130 26105-252130 26305-252130 25105-252130 26505-252130 3.0 25005-253030 25205-253030 25305-253030 25405-253030 25505-253030 25905-253030 25805-253030 25705-253030 26105-253030 26305-253030 25105-253030 26505-253030 4.0 25005-254030 25205-254030 25305-254030 25405-254030 25505-254030 25905-254030 25805-254030 25705-254030 26105-254030 26305-254030 25105-254030 26505-254030 4.6 25005-254630 25205-254630 25305-254630 25405-254630 25505-254630 25905-254630 25805-254630 25705-254630 26105-254630 26305-254630 25105-254630 26505-254630

www.thermoscienti c.com/chromatography 4-039 4 -040 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Hypersil GOLD Ordering Guide GOLD Hypersil Preparative Guard Cartridge Holder Guard Cartridge Preparative Format (µm) Size Particle (µm) Size Particle Holder Cartridge Guard UNIGUARD Format 1.9 5 5 12 Format Format Column (3/pk) Column HTS Javelin Column (3/pk) Column HTS Javelin Cartridge (3/pk) Cartridge Guard Preparative Column Preparative HPLC Cartridge (3/pk) Cartridge Guard Preparative Column Preparative HPLC 10 (mm) Length 10 (mm) Length Length (mm) ID (mm) ID (mm) Length (mm) ID (mm) Length 10 20 10 50 100 150 250 10 50 100 150 250 continued 20 10 (mm) ID 4.0/4.6 3.0 2.1 1.0 (mm) ID 2.1 4.0 10 20 10 21 30 50 10 21 30 50 10 21 30 50 10 21 30 50 10 20 10 21 30 50 10 21 30 50 10 21 30 50 10 21 30 50 854-00 C-1000 No. Cat. 850-00 852-00 852-00 851-00 No. Cat. GOLD GOLD 25002-012135 25005-024035 50-103235092 25405-019023 25305-019023 25005-019023 50-123235092 25405-019223 25305-019223 25005-019223 50-500235097 25405-059070 25305-059070 25005-059070 50-520235097 25405-059270 25305-059270 25005-059270 50-530235097 25405-059370 25305-059370 25005-059370 50-550235097 25405-059570 25305-059570 25005-059570 25005-109070 50-020235197 25405-109270 25305-109270 25005-109270 50-030235197 25405-109370 25305-109370 25005-109370 50-050235197 25405-109570 25305-109570 25005-109570 50-500235197 25405-159070 25305-159070 25005-159070 50-520235197 25405-159270 25305-159270 25005-159270 50-530235197 25405-159370 25305-159370 25005-159370 50-550235197 25405-159570 25305-159570 25005-159570 50-500235297 25405-259070 25305-259070 25005-259070 50-520235297 25405-259270 25305-259270 25005-259270 50-530235297 25405-259370 25305-259370 25005-259370 50-550235297 25405-259570 25305-259570 25005-259570 25012-019023 25012-019223 25012-059070 25012-059270 25012-059370 25012-059570 25012-109070 25012-109270 25012-109370 25012-109570 25012-159070 25012-159270 25012-159370 25012-159570 25012-259070 25012-259270 25012-259370 25012-259570 aQ 50-00025405-109070 25305-109070 – – – – – – – – – – – – – – – – – – PFP – – – – – – – – – – – – – – – – – – Notes LC Columns and Accessories

>> Columns for Analytical LC

www.thermoscienti c.com/chromatography 4-041 4 -042 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories phase bondingandpacking,ThermoScientific Technology Founded onstate-of-the-artCoreEnhanced Ultimate CorePerformance–SpeedandSelectivityCombined Thermo Scienti c Accucore HPLCColumns on investment. these columnsprovideanexcellentreturn and compatiblewithalmostanyinstrument, range ofstationaryphaseselectivities workflow andefficiency. Available inawide chromatography solutiontoenhancelaboratory Accucore highest quality that allcolumnsarepackedwiththe Enhanced automatedproceduresensure Automated PackingProcess excessive backpressure high resolutionseparationswithout layer, theseparticlesgeneratehighspeed, With asolidcentralcoreandporousouter Solid CoreParticles ™ HPLC columns provide a unique HPLCcolumnsprovideaunique ™ and utilizing vast experience in andutilizingvastexperiencein and applicationnotes productView information The key components of Core Enhanced Technology Core of keyThe components ™

selectivities tomeetallyourseparationneeds. rugged columnsthatofferawiderangeof automated processesresultinhighlyreproducible, technology andpackedwithhighlycontrolled different stationaryphasesbondedusingadvanced lower thanthoseassociatedwithUHPLC.Eleven separation, withbackpressuressignificantly columns allowshighspeed,resolution narrow particlesizedistribution;AccucoreHPLC engineered toadiameterof2.6μmandvery Containing solidcoreparticles,whichare www.thermoscienti c.com/accucore Visit itat: for theAccucoreHPLCcolumnrange. latest news,applicationsanddownloads The Accucorewebpagecontainsthe of highcoverage,robustphases Optimized phasebondingcreatesaseries Advanced BondingTechnology high efficiencycolumns size distributiontoaminimumandproduces Enhanced selectionprocesskeepsparticle Tight ControlofParticleDiameter LC Columns and Accessories

18.0 18.0 fully porous, 5µm 16.0 16.0 fully porous, 3µm 14.0 14.0 fully porous, <2µm 12.0 fully porous, <2µm 12.0 Decreasing Ef ciency Decreasing Ef 10.0 Accucore RP-MS 2.6µm

H 10.0 H 8.0 8.0 6.0 6.0 4.0 4.0 2.0 2.0 0.0 0.00.0 2.0 4.0 6.0 8.0 10.0 0.0 2.0 4.0 6.0 8.0 10.0 Accucore HPLC columns are more µ (mm/s) ef cient than 5μm and 3μm columns.

fully porous, 5µm

fully porous, 5µm 1000 1000

900 fully porous, 3µm >> 900 fully porous, 3µm 800 800 fully porous, <2µm Columns for Analytical LC 700 fully porous, <2µm 700

Increasing Pressure 600 Accucore RP-MS 2.6µm 600 Accucore RP-MS 2.6µm 500 500 Typical HPLC Pressure Limit 400 Typical HPLC Pressure Limit Pressure (bar) 400 Pressure (bar) 300 Column dimensions: 100 x 2.1mn 300 Column dimensions: 100 x 2.1mn 200 200 100 100 0 0 0 200 400 600 800 1000 0 200 400 600 800 1000 Accucore HPLC columns achieve this Flow Rate (µL/min) ef ciency without excessive backpressure

Accucore 2.6µm HPLC Columns Optimum Conditions and Ratings Column ID (mm) Optimum Flow Rate Maximum Inj. Volume Backpressure Rating Temperature Rating 2.1 400µL/min 1µL 1000 bar 70°C 3.0 800µL/min 3µL 1000 bar 70°C 4.6 1800µL/min 5µL 1000 bar 70°C

www.thermoscienti c.com/chromatography 4-043 4 -044 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories The selectivityofferedbyAccucoreRP-MSmatchesthatofC18columns. and lowpeaktailingmakethisthephaseofchoiceforusewithMSdetection. RP-MS offersslightlylowerretentionthanC18andthiscombinedwithhighefficiencies interactions andthushighlyefficientpeakswithverylowtailing. the silicasurface.Thiscoverageresultsinasignificantreductionnon-hydrophobic Accucore RP-MSusesanoptimizedalkylchainlengthformoreeffectivecoverage of • • • Accucore RP-MS

mAU mAU Excellent combinationofspeedandef ciency Excellent peakshapes Optimized forMSdetection 120 160 200 240 100 200 300 400 500 600 40 80 0 0 0.0 Bases 0 Testosterones 1 2 0.5 3 0.5 1 4 1.0 2 Minutes 1 1.5 Minutes 5 2.0 1.5 3 6 2.5 Tf =1.27 Tf 2 3.0 bar 232 1.Uracil 215nm at Analytes: UV pressure: Back 1µL 500µL/min Injection volume: Detection: Temperature: 30°C 25mM /35% rate: Flow Methanol 65% phase: Mobile x2.1mm 50mm 2.6µm, RP-MS Accucore nlts 1. 11-Ketotestosterone 254nm at UV Analytes: 1µL 0.6mL/min Injection volume: Detection: /Acetonitrile Temperature: Water (v/v) 40°C 60:40 rate: Flow phase: Mobile x2.1mm 100mm 2.6µm, RP-MS Accucore 2.5 6. Amitriptyline 5. Acenaphthene 4. Naphthalene 3. Butylparaben 2. Propranolol pH7.0Potassium Phosphate 3. Epitestosterone (Nandrolone)2. 19-Nortestosterone Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 Low 02 01 2.6µm 80Å 2 to9 7 High 300 5 4 LC Columns and Accessories Accucore C18 Hydrophobicity • Optimum retention of non-polar compounds Low High pH Range • Hydrophobic interaction mechanism • Separates a broad range of analytes 011 to 11 4 Pore Size

The carbon loading of Accucore C18 phase provides high retention of non-polar 0 80Å 300 analytes via a predominantly hydrophobic interaction mechanism. Carbon Load (%) The highly retentive nature of Accucore C18 phase means that it can be used 029 5 to separate a broad range of analytes. Particle Size 2.6µm USP L1

Triazines

100 Accucore C18 2.6µm, 50mm x 2.1mm 3 Mobile phase: A – Water ; B – Acetonitrile Gradient: Time (min) %B 80 1.0 35 2 2.5 70

4 6 Flow rate: 600µL/min 60 Temperature: 25°C 1 Detection: UV at 280nm >> mAU Injection volume: 2µL Columns for Analytical LC 40 Backpressure: 298 bar 5 Analytes: 1. Simazine 2. Simetryn 3. Atrazine 20 4. Ametryn 5. Propazine 6. Prometryn 0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 Minutes

www.thermoscienti c.com/chromatography 4-045 222 2 4 -046 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories HPLC columnstobenefitfromexcellentpeakshapes. are theresultofexcellentbondedphasecoverageandallowusersAccucoreC8 The lowlevelsofsecondaryinteractionsdemonstratedinthephasecharacterization phase providesoptimumretention. recommended foranalyteswithmediumhydrophobicityorwhenalesshydrophobic with longeralkylchainlengthmaterial,suchasC18,andaretherefore Accucore C8HPLCcolumnsofferlowerhydrophobicretentionthanpacked • • • • Accucore C8

Relative Abundance Recommended formoderatelypolaranalytes Low levelsofsecondaryinteractions Complementary stericselectivitytoC18 Lower hydrophobicretention 10 40 50 60 70 80 90 20 30 10 0 0 0. Testosterone 00 Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low Testosterone .2 5 80 Å 2.6µm 0. 4 2 to9 0. 6 Minutes 0. 8 1. 0 USP L7 1. High 300 2 14 25 1. 4 222 2 Data fromsixinjections Mobile phaseB: Mobile phaseA: Column Detection: Injection: Te Flow: Gradient: mperature:

:

Retention time(t

%RSD Area eeto: ESI-MS/MS%RSD Peak area Detection: °C 60 Injection: 5μL Temperature: Flow: Gradient: Mobile phase B: Mobile phase A: x2.1mm 50 2.6μm, C8 Accucore Data fromsixinjections.

Relative Abundance %RSD t 10

70 80 90 20 30 40 50 60 10 water +0.1%formicacid Accucore C82.6µm,50x2.1mm ESI-MS/M 5 µL 60 °C 1500 µL/min 5–95 %Bin0.8minutes acetonitrile +0.1%formicacid Retention time (tR /min) (tR time Retention 0 0

0. 00 R Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low Testosterone %RSD Area R 5–95 % B in 0.8 minutes 0.8 %Bin 5–95 %RSD tR %RSD /min)0 S . acetonitrile + 0.1% formic acid +0.1% formic acetonitrile water + 0.1% formic acid +0.1% formic water .2 1500 μL/min 1500 5 80 Å 2.6µm 0. 4

2 to9 0.22 3.01 0.73 0. 6 0.22 3.01 Minutes .73 0. 8 1. 0 USP L7 1. High 300 2 14 25 1. 4 Data fromsixinjections Column Detection: Injection: Te Flow: Gradient: Mobile phaseB: Mobile phaseA: mperature:

:

Retention time(t

%RSD Area

%RSD t

ESI-MS/M 5 µL 60 °C 1500 µL/min 5–95 %Bin0.8minutes acetonitrile +0.1%formicacid water +0.1%formicacid Accucore C82.6µm,50x2.1mm R R /min)0 S .

0.22 3.01 .73 LC Columns and Accessories Hydrophobicity Accucore aQ Hydrophobicity

Low High • Retention and resolution of polar analytes Low High pH Range pH Range • Polar endcapped C18 stationary phase for alternative selectivity

0 2 to 9 14 • Ideal for highly aqueous mobile phases 012 to 9 4 Pore Size Pore Size

0 80 Å 300 The polar functional group used to endcap Accucore aQ phase provides an 0 80Å 300 Carbon Load (% ) additional controlled interaction mechanism by which polar compounds can be Carbon Load (%) retained and resolved, making Accucore aQ phase ideal for the quantitative 0 5 25 analysis of trace levels of polar analytes. 029 5 Particle Size 2.6µm USP L1 Particle Size 2.6µm USP L7 The wettability of reversed phase media can be increased by the introduction of polar functional groups. The polar endcapping of Accucore aQ media also makes it usable in 100% aqueous mobile phases without the risk of loss of performance or poor stability.

Testosterone Lamivudine (USP) >> 100 Column: Accucore C8 2.6 µm,140 50 x 2.1 mm Accucore aQ 2.6µm, 50mm x 2.1mm 90 Mobile phase A: water + 0.1% formic acid Mobile phase: 95:5 (v/v) Ammonium Acetate, Columns for Analytical LC 120 pH 3.80 / Methanol 80 Mobile phase B: acetonitrile + 0.1% formic acid Flow rate: 200µL/min 70 Gradient: 5–95 % B in 0.8 minutes100 Temperature: 35°C 60 Flow: 1500 µL/min Detection: UV at 277nm

mAU 80 50 Temperature: 60 °C Injection volume: 1µL Analytes: Lamivudine 40 Injection: 5 µL 60 Asymmetry 1.36

30 Detection: ESI-MS/MS %RSD tr 0.00 Relative Abundance 40 20 %RSD Peak area 1.72 (%RSD calculated from 6 replicate injections) 10 20 USP acceptance criteria: % RSD (t , Peak Area) <2.0 Retention time (t /min)0.73 r 0 R %RSD t 0.22 0.00.2 0.4 0.6 0.8 1.0 1.2 1.4 R 0 %RSD Area 3.01 Minutes 0.0 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 Data from six injections. Minutes

2222

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4 -048 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories

mAU undergo phasecollapsein100%aqueousmobilephase. The speciallydesignedbondedphaseisstablefrompH1.5to10.5andwillnot complementary tostandardC18phases. material thatoffershighefficiency, wideroperatingpHrangeanduniqueselectivity Accucore PolarPremiumisanexceptionallyruggedpolarembeddedreversephase • • • Accucore Polar Premium 600 100 200 300 400 500 Stable overawidepHrangeandcompatiblewith100%aqueous Selectivity complementarytoconventionalC18phases Rugged amide-embeddedC18phase -5 mobile phase 0 Particle Size Carbon Load(% ) Pore Size pH Range H 0 0 0 Low 0.00 y Curcuminoids (Tumeric) d rop h o bi c i ty 2.6 1,2,3 1, µ 2, m 8 3 1.5 to10 1.00 1 150 Å Minutes 2 Fused coreC18 2.00 3 USP L60 Ac cuco High 300 14 25 re Fu

Polar se d Co

Premium re

C1 3.00 8

while theC18columnsfailtoseparatethesecomponents. under simpleisocraticconditionsinlessthanthreeminutes, selectivity thatresolvesthemajorandminorcomponent The AccucorePolarPremiumHPLCcolumnprovides nlts 1. Curcumin 428nm at UV Analytes: Detection: °C 40 Injection: 6μL Temperature: Flow: Mobile phase: mm x3.0 100 C18, Core Fused x3.0mm 100 2.6μm, Premium Polar Accucore

methanol : 10mM phosphoric acid, 80 :20 80 acid, phosphoric :10mM methanol 800 μL/min 800 3. Bis-desmethoxycurcumin 2. Desmethoxycurcumin mAU 100 200 300 400 500 600 -5 0 Particle Size Carbon Load(% ) Pore Size pH Range H 0 0 0 Low 0.00 y d rop h o bi c i ty 2.6 1,2,3 1, µ 2, m 8 3 1.5 to10 1.00 1 150 Å desirable Minutes 2 Fused coreC18 2.00 3 USP L60 Ac cuco High 300 14 25 re Fu

Polar se d Co

Premium re

C1 3.00 8

LC Columns and Accessories Accucore Phenyl-Hexyl Hydrophobicity • Mixed-mode selectivity for aromatic and moderately polar analytes Low High pH Range • Enhanced Pi-pi interactions with aromatics • Moderate hydrophobicity 012 to 8 4 Pore Size

The C6 chain in Accucore Phenyl-Hexyl phase exhibits classical RP retention 0 80Å 300 and selectivity, while the phenyl ring can add special selectivity by interacting Carbon Load (%) with polar groups within the solutes. This results in a mixed-mode separation mechanism. The reduced hydrophobicity of this phase makes it ideal for the 025 5 separation of very non-polar compounds. Particle Size 2.6µm USP L11 The Phenyl-Hexyl phase should be selected for complex samples where some peaks are well resolved on a conventional alkyl phases, but are not well resolved on a conventional phenyl phase, or when other peaks are well resolved on a phenyl phase, but not well resolved on a conventional alkyl phase.

Beta-agonists >>

Columns for Analytical LC 100 2.89 NL: 9.22E5 Accucore Phenyl-Hexyl 2.6µm, 100mm x 2.1mm m/z = 225.60-226.60 Mobile phase: A – Ammonium acetate 5mM, pH 4 Terbutaline B – Acetonitrile 0 Gradient: Time (min) %B 100 3.04 NL: 6.51E5 0 5 m/z = 239.60-240.60 MS 1 5 Cimaterol 10 100 0 Flow rate: 0.25mL/min 100 NL: 3.52E6 3.16 Temperature: 40°C m/z = 219.60-220.60 Detection: +ESI-MS (45ºC, 4.5kV, 60V, Salbutamol scan 150 – 350) 0 Injection volume: 1µL

100 NL: 2.23E6 4.72 Backpressure: 120 bar (at t0) m/z = 301.60-302.60 MS Ractopamine 0

100 NL: 2.51E6 4.80 m/z = 213.60-214.60 MS Clorprenaline 0 100 NL: 1.71E6 5.13 m/z = 276.60-277.60 MS Clenbuterol 0 0 1 2 3 4 5 6 7 Minutes

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mAU mAU mobile phasesandrobust,demonstratingverylowbleed. The advanceddesignofthebondedphasemakesitcompatiblewithhighlyaqueous Phenyl-X exhibitsparticularlyhigharomaticselectivity. selectivity whencomparedtootherreversedphasematerialssuchasC18orPhenyl. The proprietaryAccucorePhenyl-Xalkylaromaticbondedphaseprovidesaunique • • • • Phenyl-X Accucore 10 12 15 10 12 Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low 25 50 75 20 40 60 80 0 5 0 0 0 0 0 Robust, high-ef ciency, lowcolumnbleed Compatible withhighlyaqueousmobilephases Enhanced selectivityforaromaticcompounds Unique reversed-phaseshapeselectivity 0. 0. Estrogens 000 00 0. 51. .5 51. .5 5 1 6 80 Å 2.6µm 1. 1 02 0 02 2 to8 2. 52. .5 52.02 .5 5 2. 2 0 02 3 Minutes 4 Minutes 2. 53. .5 2 53. .5 5 3 3. 0 03 03 4 2.04 3. 54.05 .5 54. .5 5 High 4.0 300 Accucore Phenyl-X 05 14 25 Fused CoreC18 5. .5 56. .5 5 6. 6. 0 0 0 Detection: UV at 220nm at phase UV mobile as Same solvent: Wash Detection: °C 40 Injection: 1μL Temperature: Flow: Mobile phase: x2.1mm 100 C18, Core Fused x2.1mm 100 2.6μm, Phenyl-X Accucore

3. Estrone(E1) 1. Estriol(E3) HO HO

HH

HH H H 15:40:45 (v/v) acetonitrile: methanol :water methanol acetonitrile: (v/v) 15:40:45

3 C H H 3 OH C 400 μL/min 400 OH OH 4. Ethynylestradiol 2. Estradiol(E2) HO HO

mAU mAU 10 12 15 10 12 HH Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low 25 50 75 20 40 60 80 0 5 0 0 0 0 0 H HH H 0. 0. 3 C 000 00 H H

OH 3 C OH CH 0. 51. .5 51. .5 5 1 6 80 Å 2.6µm 1. 1 02 0 02 2 to8 2. 52. .5 52.02 .5 5 2. 2 0 02 3 Minutes 4 Minutes 2. 53. .5 2 53. .5 5 3 3. 0 03 03 4 2.04 3. 54.05 .5 54. .5 5 High 4.0 300 Accucore Phenyl-X 05 14 25 Fused CoreC18 5. .5 56. .5 5 6. 6. 0 0 0

3. Estrone(E1) 1. Estriol(E3) HO HO HH

HH H H

3 C H H 3 OH C OH OH 4. Ethynylestradiol 2. Estradiol(E2) HO HO

HH H HH H 3 C H H

OH 3 C OH CH LC Columns and Accessories Accucore PFP Hydrophobicity • Alternative selectivity to C18 Low High pH Range • Extra retention for halogenated species • Unique selectivity for non-halogenated polar compounds 012 to 8 4 Pore Size

The introduction of fluorine groups into the Accucore PFP (pentafluorophenyl) 0 80Å 300 stationary phase causes significant changes in solute-stationary phase interactions. Carbon Load (%) This can lead to extra retention and selectivity for positional isomers of halogenated compounds. 025 5 Particle Size 2.6µm USP L43 PFP Columns are also well suited to the selective analysis of non-halogenated compounds, in particular polar compounds containing hydroxyl, carboxyl, nitro, or other polar groups. High selectivity is often most apparent when the functional groups are located on an aromatic or other rigid ring system.

Positional isomers

50 Accucore PFP 2.6µm, 50mm x 2.1mm >> 5 Mobile phase: A – 0.1% Formic Acid in Water Mobile phase: A – 0.1% Formic Acid in Acetonitrile Columns for Analytical LC 40 Gradient: 15-30%B in 7 minutes Flow rate: 600µL/min 7 Temperature: 50°C Detection: UV at 270nm 30 1 Injection volume: 2µL 8 9 1. 3,4 – Dimethoxyphenol 10 2. 2,6 – Dimethoxyphenol 20 mAU 2 12 3. 2,6 – Di uorophenol 11 4. 3,5 – Dimethoxyphenol 13 14 5. 2,4 – Di uorophenol 6 6. 2,3 – Di uorophenol 10 3 4 7. 3,4 – Di uorophenol 8. 3,5 – Dimethylphenol 9. 2,6 – Dimethylphenol 0 10. 2,6 – Dichlorophenol 11. 4 – Chloro-3-Methylphenol 12. 4 – Chloro-2-Methylphenol 0 1 2 3 54 6 7 8 13. 3,4 – Dichlorophenol Minutes 14. 3,5 – Dichlorophenol

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28

4 -052 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories

mAU particle resultinaphasethatisstableevenhighlyaqueousmobilephases. optimized bondingdensityofthelongalkylchainsfacilitatedbyawiderporediameter It isalsoanexcellentalternativetonormal-phasecolumnsforlipidanalysis.The of shapeselectivityfromthatmeasuredintheSSphasecharacterisationtest. related isomers,forexamplecarotenoidsandsteroids.Thisisadifferentform Accucore C30offershighshapeselectivityforhydrophobic,longchain,structurally • • • Accucore C30 Excellent aqueous-compatibility Excellent High shapeselectivityforstructurallyrelatedisomers Ideal forseparationofhydrophobic,longalkylchaincompounds 10 15 20 25 Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low -5 50 0 0 0 0 0 0 0.00 Vitamin Kisomers 5 1.00 2.6µm 2 to8 2.00 150 Å 1 Minutes 1 3.00

4.00 USP L62 2+ 2’

High

300 14 25 5.00 Fu Ac 2 se cuco d 2’ Co

re re

C3

C1 6.00 0 8

Minutes 1-Vitamin K2, 2-Vitamin K1 (trans isomer), 2’-Vitamin K1 (cis isomer) K1 (cis 2’-Vitamin isomer), K1 (trans 2-Vitamin K2, 1-Vitamin Minutes Kcompounds Vitamin of separation the showing Chromatogram 250nm at UV the C18 than column. K1 isomers vitamin for separation better shows C30 Accucore Detection: °C 20 Injection: 5μL Temperature: Flow: Mobile phase: x3.0mm 100 C18, Core Fused x3.0mm 100 2.6μm, C30 Accucore 28

650μL/min methanol: 2mM ammonium acetate, 98:2 acetate, ammonium 2mM methanol:

mAU 10 15 20 25 Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low -5 50 0 0 0 0 0 0 0.00 5

1.00 2.6µm 2 to8 2.00 150 Å 1 Minutes 1 3.00

4.00 USP L62 2+ 2’

High

300 14 25 5.00 Fu Ac 2 se cuco d 2’ Co

re re

C3

C1 6.00 0 8

LC Columns and Accessories Accucore HILIC Hydrophobicity • Enhanced retention of polar and hydrophilic analytes Low Very Low (5%) High pH Range • Alternative selectivity to C18 without ion-pair or derivatization

012 to 8 4 Pore Size In HILIC mode the separation occurs through two mechanisms. The primary mechanism is a partitioning effect due to the enriched water layer around the polar 0 80Å 300 or charged substrate material. The secondary mechanism involves interaction between the analyte and the active surface moiety. Particle Size 2.6µm USP L3 Analyte properties that govern retention with HILIC phases are acidity/basicity, which determines hydrogen bonding, and polarizability which determines dipole- dipole interactions. The highly organic mobile phases used with Accucore HILIC phase ensure efficient desolvation in ESI MS detection, which in turn leads to improved sensitivity.

Catecholamines

2 >> Accucore HILIC 2.6µm, 50mm x 2.1mm

140 Mobile phase: 85:15 Acetonitrile:100mM Columns for Analytical LC Ammonium Formate, pH 3.2 120 Flow rate: 2mL/min Temperature: 40°C 1 Detection: UV at 280nm 100 4 Injection volume: 5µL Backpressure: 157 bar 80 Analytes: 1. Catechol 3 2. 5-HIAA mAU 60 3. DOPAC 4. Serotonin 5. L-tyrosine 40 5 6. Dopamine 7. L-DOPA 6 20 7

0 0 0.5 1 1.5 2 2.5 3 3.5 4 Minutes

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mAU 20 40 60 analytes usingupto20%aqueousmobilephase. The bondedhydrophilicstationaryphaseprovidesretentionofbroadrangepolar than otherHILICphases. Accucore Urea-HILIChasanalternativeselectivityandlowerionexchangeactivity • • • Accucore Urea-HILIC 0 0 0 0 Low ionexchangeactivity Unique selectivitycomparedtootherHILICphases Bonded hydrophilicstationaryphase 0 Analgesic compounds Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 0. 2 80 Å 2.6µm 0. 4 2 to8 0. 6 Minutes 1 0. 8 2 1.

0 3 1. 2 High 300 14 1. 4

Retention time (t acid salicylic and aspirin acetaminophen, of amixture of analyses replicate eight from Data Detection: UV at 230 nm 230 at UV bar 71 water:acetonitrile 20:80 Backpressure: mode. loop partial 10 μL into 2μL Detection: solvent: wash 2minutes Injection °C 35 Injection: Temperature: Time: Run Flow: Mobile phase: x2.1mm 100 2.6μm, Urea-HILIC Accucore CV % CV Mean

Acetaminophen 0.760 0.00 t R composition 10:80:10, A:BC 10:80:10, composition R ), peak asymmetry (A asymmetry peak ), 1.474 300 μL/min 300 C: 100 mM ammonium acetate adjusted to pH 4.9 pH to adjusted acetate ammonium mM 100 C: B: acetonitrile water A: 1.17 A s 0.908 .809 0.49 0.92 0.48 mAU t 20 40 60 R 0 0 0 0 Salicylic acid Salicylic 0 s Analgesic compounds ), peak resolution (R resolution peak ), Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 1.303 A s 2.359 0. R 2 s 1.100 0.00 80 Å 2.6µm s t ) R 0. 4 2 to8

Aspirin 1.318 .30.48 0.63 A s 0. 3.264 6 R s Minutes 1 0. 8 2 1.

0 3 1. 2 High 300 14 1. 4

Accucore XL HPLC Columns LC Columns and Accessories

Based on Core Enhanced Technology using 4μm solid core particles, Accucore XL HPLC columns allow users of conventional HPLC methods to enjoy performance far beyond that of columns packed with 5μm, 4μm or even 3μm fully porous particles. Very high separation efficiencies using standard HPLC instruments and conditions provide increased peak resolution and lower limits of detection. An ultra-stable packed bed results in exceptionally robust columns that demonstrate excellent retention and response reproducibility.

4µm Solid Core Particles for all Users The 4µm solid core particles used in Accucore XL HPLC columns have been specifically designed to get the optimum chromatographic performance from conventional HPLC instruments.

• Very high ef ciencies

• Little decrease in ef ciency as  ow rate is increased >>

• Moderate backpressures Columns for Analytical LC

Ef ciency Accucore XL HPLC columns generate higher efficiencies than columns packed with 5µm and 3µm fully porous material – as shown in the van Deemter curve below.

18.0 Fully porous C18, 5μm

16.0 Fully porous C18, 3μm

14.0 Accucore XL C18, 4μm

12.0 Identical instrument and method 10.0 conditions for all columns Dimensions: 150 x 4.6mm 8.0 Mobile Phase: 50% water: 50% acetonitrile Plate Height (H, µm) Temperature: 30°C 6.0 Injection Volume: 1µL

Increasing Ef ciency Detection: UV at 254nm (0.1s rise time, 20 Hz) Sample: o-xylene 4.0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.04.5 Linear velocity (u, mm/s)

• 75% higher ef ciency than 5µm fully porous • 50% higher ef ciency than 3µm fully porous

www.thermoscienti c.com/chromatography 4-055 4 -056 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories of analytes. retentive natureofthephasemeansthatitcanbeusedtoseparateabroadrange analytes viaapredominantlyhydrophobicinteractionmechanism.Thehighly The carbonloadingofAccucoreXLC18provideshighretentionnon-polar • • • Accucore C18 XL • WithinconventionalHPLCinstrumentationpressurelimit–evenathighow rates • Backpressuresbetweenthosegeneratedby3µmand5µmfullyporousparticles porous 3µm,thatarecompatiblewithconventionalHPLCinstruments. Accucore XLHPLCcolumnsgeneratereasonablebackpressures,moderatelyhigher thanfullyporous5µmandlower Backpressure Ibuprofen andValerophenone (USP) mAU mAU Separates abroadrangeofanalytes Hydrophobic interactionmechanism Optimum retentionofnon-polarcompounds Pressure (bar) 10 15 20 25 30 10 15 20 25 30 -5 -5 100 200 300 400 500 600 0 5 0 5 2.5 0 2.5 0 0 0.0 5.0 5.0 0.5 7.5 7.5 10.0 10.0 400 barconventionalHPLCpressure 12.5 12.5 1.0 Flow rate(mL/min) Minutes Minutes 15.0 15.0 1 2 17.5 17.5 S/N =1202 N =18274plates 1 1.5 20.0 20.0 22.5 22.5 2 25.0 25.0 S/N =534 N =10538plates 2. 02 27.5 27.5 30.0 30.0 nlts 1. Valerophenone 214nm at UV Analytes: 5µL 2mL/min Injection volume: Detection: Temperature: 30°C with water (v/v) rate: Flow 66.3:33.7 phase: Mobile x4.6mm 150 5µm, C18 porous Fully x4.6mm 150 4µm, C18 XL Accucore .5 eprtr: 30°C acetonitrile :50% water 50% x4.6mm 150 Temperature: Phase: Mobile Dimensions: conditions for all columns method and instrument Identical • 125%highersensitivity • 73%higheref ciency 2. Ibuprofen 2.5:methanol pH acid, phosphoric

AccucoreXLC18,4μm FullyporousC18,3μm FullyporousC18,5μm Particle Size Carbon Load(% ) Pore Size pH Range Hydrophobicity 0 0 0 Low 4µm 80Å 7 1 to11 USP L1 High 300 14 25 LC Columns and Accessories Accucore XL C8 Hydrophobicity • Similar selectivity to C18 with lower retention Low High pH Range • Recommended for analytes with moderate hydrophobicity

0 2 to 9 14 Pore Size Accucore XL C8 offers lower hydrophobic retention than columns packed with

longer alkyl chain length material, such as C18. It is then therefore recommended 0 80Å 300 for analytes with moderate hydrophobicity, or when a less hydrophobic phase Carbon Load (%) provides optimum retention. 0 4 25 Particle Size 4µm USP L7

Endocrine Disruptors

Accucore XL C8 4µm, 150 x 4.6mm 400 Fully porous C8 5µm, 150 x 4.6mm

300 Mobile phase A: water Mobile phase B: acetonitrile >> 1 Gradient: Time (min) % B 200 3 Rs = 4.12

2 0.0 25 Columns for Analytical LC mAU 5 100 4 20.0 70 20.1 75 0 25.0 25 Flow rate: 1.5mL/min 0 1 2 3 4 5 6 7 8 9 10 Minutes Temperature: 25°C Detection: UV at 220nm Injection volume: 5µL 400 1 Analytes 1. Desethyl Atrazine 2. Simazine 300 3. Atrazine 3 4. Diuron 2 200 Rs = 5.14 5. Bisphenol A 5 mAU 4 100

0 0 1 2 3 4 5 6 7 8 9 10 Minutes

• 31% better resolution of critical pair • 37% narrower peaks • 226% higher sensitivity

www.thermoscienti c.com/chromatography 4-057 4 -058 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Accucore Guide Ordering 4 (µm) Size Particle 2.6 (µm) Size Particle HPLC Column HPLC HPLC Column HPLC Drop-in Guard (4/pk) Format (4/pk) Defender Guard Format 250 150 100 250 150 100 50 30 50 10 (mm) ID (mm) Length 10 (mm) ID (mm) Length 4.6 3.0 2.1 4.6 3.0 2.1 4.6 3.0 2.1 2.1 2.1 2.1 2.1 2.1 4.6 3.0 2.1 4.6 3.0 2.1 2.1 4.6 3.0 4.6 3.0 4.6 3.0 4.6 3.0 74104-254630 74204-253030 74104-253030 74104-252130 74104-154630 74204-153030 74104-153030 74104-152130 74104-104630 74104-103030 74104-102130 – 17626-152130 17626-102130 17626-052130 17626-032130 74104-054630 74204-053030 74104-053030 74204-014001 74104-052130 74204-013001 74104-014001 74104-013001 74104-012101 C18 17626-012105 RP-MS 72-560116143 17226-154630 17326-153030 17126-154630 17326-104630 17226-153030 17626-154630 17126-153030 17226-104630 17626-153030 17326-103030 17126-104630 17226-103030 17626-104630 17326-054630 17126-103030 17626-103030 17226-054630 17326-053030 17126-054630 17226-053030 17626-054630 17126-053030 17626-053030 17326-014005 17226-014005 17126-014005 17626-014005 17626-013005 74204-254630 74204-252130 74204-154630 74204-152130 74204-104630 74204-103030 74204-102130 – 17126-152130 17126-102130 17126-052130 17126-032130 74204-054630 74204-052130 74204-012101 C8 17126-012105 C18 17126-013005 – 17326-152130 17226-152130 17326-102130 17226-102130 17326-052130 17226-052130 17226-032130 72-11517326-012105 17226-012105 C8 72-10517326-013005 17226-013005 – 17326-032130 17326-154630 aQ 82-500196133 72-500146133 72-50017526-153030 27826-153030 17426-153030 27826-103030 27926-153030 17426-103030 28026-252130 17926-153030 28026-154630 27926-103030 28026-153030 28026-152130 17926-103030 17526-012105 28026-104630 28026-103030 27826-012105 28026-102130 28026-054630 17426-012105 28026-053030 28026-052130 27926-012105 – – 17926-012105 – 28026-012105 Premium Polar 72-56027926-154630 17526-054630 – 27926-152130 17926-154630 27826-054630 17526-052130 27926-104630 17926-152130 17426-054630 27826-052130 17926-104630 27926-102130 27926-054630 17426-052130 17926-102130 27926-053030 17926-054630 27926-052130 17926-053030 17926-052130 17926-032130 17926-014005 17926-013005 Phenyl-Hexyl 27926-252130 – – – Phenyl-X – 17426-154630 17426-152130 17426-104630 17426-102130 17426-053030 17426-032130 17426-014005 17426-013005 PFP 27826-252130 27826-154630 27826-152130 17526-102130 27826-104630 27826-102130 17526-053030 27826-053030 – – – C30 – 17526-154630 17526-152130 17526-104630 17526-032130 17526-014005 17526-013005 HILIC 17526-103030 27726-252130 27726-154630 27726-152130 27726-104630 27726-102130 27726-053030 – – – Urea-HILIC 27726-153030 27726-103030 27726-054630 27726-052130 27726-012105 LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) RP-MS C18 C8 aQ Polar Premium Phenyl-Hexyl Phenyl-X PFP C30 HILIC Urea-HILIC 2.6 Defender Guard 10 2.1 17626-012105 17126-012105 17226-012105 17326-012105 28026-012105 17926-012105 27926-012105 17426-012105 27826-012105 17526-012105 27726-012105 (4/pk) 3.0 17626-013005 17126-013005 17226-013005 17326-013005 – 17926-013005 – 17426-013005 – 17526-013005 – 4.6 17626-014005 17126-014005 17226-014005 17326-014005 – 17926-014005 – 17426-014005 – 17526-014005 – HPLC Column 30 2.1 17626-032130 17126-032130 17226-032130 17326-032130 – 17926-032130 – 17426-032130 – 17526-032130 – 50 2.1 17626-052130 17126-052130 17226-052130 17326-052130 28026-052130 17926-052130 27926-052130 17426-052130 27826-052130 17526-052130 27726-052130 3.0 17626-053030 17126-053030 17226-053030 17326-053030 28026-053030 17926-053030 27926-053030 17426-053030 27826-053030 17526-053030 27726-053030 4.6 17626-054630 17126-054630 17226-054630 17326-054630 28026-054630 17926-054630 27926-054630 17426-054630 27826-054630 17526-054630 27726-054630 100 2.1 17626-102130 17126-102130 17226-102130 17326-102130 28026-102130 17926-102130 27926-102130 17426-102130 27826-102130 17526-102130 27726-102130 3.0 17626-103030 17126-103030 17226-103030 17326-103030 28026-103030 17926-103030 27926-103030 17426-103030 27826-103030 17526-103030 27726-103030 4.6 17626-104630 17126-104630 17226-104630 17326-104630 28026-104630 17926-104630 27926-104630 17426-104630 27826-104630 17526-104630 27726-104630 150 2.1 17626-152130 17126-152130 17226-152130 17326-152130 28026-152130 17926-152130 27926-152130 17426-152130 27826-152130 17526-152130 27726-152130 3.0 17626-153030 17126-153030 17226-153030 17326-153030 28026-153030 17926-153030 27926-153030 17426-153030 27826-153030 17526-153030 27726-153030 4.6 17626-154630 17126-154630 17226-154630 17326-154630 28026-154630 17926-154630 27926-154630 17426-154630 27826-154630 17526-154630 27726-154630

250 2.1 – – – – 28026-252130 – 27926-252130 – 27826-252130 – 27726-252130 >> Columns for Analytical LC

Format Length (mm) ID (mm) Cat. No. 10 2.1 852-00 UNIGUARD Guard Cartridge Holder 3.0 852-00 4.6 850-00

Particle Size Length ID Format Validation Narrow Selectivity Wide Selectivity Polar Selectivity (µm) (mm) (mm) 2.6 50 2.1 3-column Kit 17126-052130-3V 17X26-052130-3VA 17X26-052130-3VB 17X26-052130-3VC 100 17126-102130-3V 17X26-102130-3VA 17X26-102130-3VB 17X26-102130-3VC 150 17126-152130-3V 17X26-152130-3VA 17X26-152130-3VB 17X26-152130-3VC Kit contains C18 C18 C18 aQ C18 RP-MS Phenyl-Hexyl PFP C18 aQ PFP HILIC

www.thermoscienti c.com/chromatography 4-059 4 -060 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories the actualchromatograms. test protocols,listthespecifications,andshow validation andcolumnperformanceexplainthe The qualityassurancereportsforsilicalot ensure thesamegreatperformanceeverytime. processes. Eachpackedcolumnistestedto stainless steelhardwareusinghighlyreliable silica ispackedinprecision-polished316 tests appropriatetoitsintendeduse.Thebonded of bondedsilicareceivesafullsuitevalidation unexpected changesinperformance.Eachbatch processes arecleanandrepeatablewithno of physicalparameters.Bydesign,thebonding only carefullyselectedlotswithnarrowranges starts withanultrapuresilicasubstrate,using can havefullconfidenceinthem.Manufacturing tested individually, sothatchromatographers analysis. TheAcclaimcolumnsarethoroughly Quality andreliabilityareessentialtoasuccessful Reliability andDurability Thermo Scientific • • • • • Optimal selectivitythroughinnovativechemistry Thermo Scienti c AcclaimHPLCColumns for usewithUHPLCsystems. with aparticlesizeof2.2μm,aredesigned phases areavailable.AcclaimRSLCcolumns, General reversed-phase,HILICandSpecialty beverage, chemical,andconsumerproducts. pharmaceutical, environmental,foodand as methods andareusedinapplicationssuch high standardssetbymodernHPLCandLC-MS symmetrical peaks.Acclaimcolumnsmeetthe selectivity,and highcolumnefficiencies, technologies. Thisprovidescomplementary advanced andinnovativecolumnbonding on high-purity, porous silicaparticles,with Reproducibleandreliablemanufacturingprocess Lowsilanolactivityforgoodbasicanalytepeakshapes Highef ciencies Ultrapure,porous,sphericalsilica Novelandproprietarysurfacechemistriesfordiversi edselectivities ™ Acclaim ™ columns are based columnsarebased enhanced hydrolyticstability. embedded reversed-phasecolumns with Acclaim PolarAdvantageII:Amide- variety ofanalytes. embedded columnforseparatingawide Acclaim PolarAdvantage:Sulfonamide- structurally relatedisomers. selectivity forseparationofhydrophobic Acclaim C30:Designedtoprovidehighshape compounds withenhancedhydrolyticstability. column forthesuperiorseparationofaromatic Acclaim Phenyl-1:Auniquereversed-phase reversed-phase column. Acclaim 120C8:High-densitymonolayerC8 resolution inavarietyofapplications. C18 reversed-phasecolumnsforexceptional Acclaim 120C18:High-density, monolayer Reversed-phase Columns provides high-performanceseparations. ensure thateachlotofbondedsilica to application-tested fortheirspecificanalysis, for bases.Thespecialtycolumnsarealso column hydrophobicity, andlowsilanolactivity contamination, stericselectivity, columnpolarity, surface coverageofthebondedphase,metal a numberofperformanceindicators,including products arethoroughlycharacterizedusing To ensureoptimalperformance,allAcclaim efficiencies forabroadrangeofanalytes. chemistries deliverexceptionalpeak laboratories today. Theinnovativesurface the highqualitystandardneededin Acclaim columnshavebeendesignedtomeet Performance Indicators of 100to1,000,000Daltons. polymers andoligomersintheMWrange designed forseparatingwatersoluble Acclaim SECColumns:Polymericcolumns Size-exclusion Columns and cationexchangecombinedinasinglecolumn. Acclaim Mixed-ModeWCX-1:Reversed-phase phase andweakanionexchangeproperties. monolayer thatincorporatesbothreversed- Acclaim Mixed-ModeWAX-1: High-density liquid chromatography(HILIC)properties. both reversed-phaseandhydrophilicinteraction Acclaim Mixed-ModeHILIC-1:Combines bases, zwitterionsandneutralmolecules. influence theseparationselectivityofacids, pH, ionicstrength,ororganicmodifierto adjustable selectivitytool,usingvariationin Mixed-mode columnsprovideaunique, Mixed-Mode Columns (see SpecialtyColumns). flexibility inadjustingselectivity chromatographic performanceandmaximum cation exchangecapabilityforunparalleled simultaneous reversed-phase,anionand Unique trimodalsurfacechemistryprovides Acclaim Trinity P1andP2columns: hydrophilic compounds Acclaim HILIC-10:Designedforseparating Hydrophilic InteractionColumns LC Columns and Accessories Acclaim 120 C18 Hydrophobicity

High performance reversed-phase columns for reproducible results Low High pH Range

0 2 to 8 14 • High hydrophobic retention Pore Size • Excellent ef ciencies for maximum resolution • Low silanol activity for excellent peak shapes for basic analytes 0 120 300 Carbon Load (%) • Reproducible manufacturing practices for reproducible column-to-column performance 0 18 25 • Extremely low bleed, fully compatible with MS Particle Size 2.2μm, 3μm, 5μm USP L1 • Available in 2.2, 3 and 5μm particle size

The Acclaim 120 columns are for high resolution reversed-phase separations. The very high surface coverage and very low metal content together result in columns with excellent efficiencies. These

columns provide exceptional performance for a variety of applications in the pharmaceutical, chemical,

environmental, and food separations areas. >> Columns for Analytical LC

160 250 220 Column: A: A: AcclaimAcclaim 120 C18,C18, 5µm, 5 µm, Detector: UV, 254 nm, 4 A 4 B 4 C 4.6 × 150mm 4.6 × 150B, C: Acclaim mm RSLC C18, .2µm, A: 1 Hz data rate B,2.1 × 50mm C: Acclaim RSLC C18, 2 µm, B: 5 Hz data rate Mobile Phase: 2.1 × 50 200mM HOAc mm C: 10 Hz data rate Mobile Phase: 200in 10% mM (v/v) HOAc MeOH Flow: A: 1.00mL/min Peaks: 1. p-Hydroxybenzoic acid

mAU inB: 10%0.41mL/min (v/v) MeOH 2. p-Hydroxybenzaldehyde 3 3 Standard 2 RSLC Column 3 Double Velocity Flow: A:C: 0.82mL/min1.00 mL/min 1 2 Column 1 6× Acceleration 2 12× Acceleration 3. Vanillic acid 1 Temperature: B: 20°C 0.41 mL/min 4. Vanillin Injection: C: A: 0.8210µL mL/min B: 1.2µL Sample: Commercial vanilla extract in 0 Temperature: 20C: 1.2µL°C 0 40% ethanol, ltered 0 Injection:Detector: A: UV, 10 254 µL nm, A: 1 Hz data rate Reference: AOAC Ofcial Method 990.25 0 10 20 30 40 50 60 0 2 4 6 8 10 0 1 2 3 4 5 B:B: 1.2 5 Hz µL data rate Minutes 25400 C:C: 1.210 Hz µL data rate Peaks: 1. p-Hydroxybenzoic acid 2. p-Hydroxybenzaldehyde 3. Vanillic acid 4. Vanillin Sample: Commercial vanilla extract in 40% ethanol, ltered Reference: AOAC Of cial Method 990.25 mAU

0

0.0 www.thermoscienti c.com/chromatography 4-061 4 -062 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 80

0 mAU pharmaceutical, environmental,foodandmanyotherindustrialsectors. activity. Thesecolumnsprovideexceptionalperformanceforavarietyofapplications inthe LC/MS compatibleC8phaseswithveryhighsurfacecoverageandextremelylowsilanol a high-purity, sphericalporoussilicasubstrate.Thecolumnsareawell-characterizedlineof Acclaim 120C8reversed-phasecolumnsfeaturedenselybondedmonolayer ligandson • • • • • hydrophobic retention High performancereversed-phasecolumnswithintermediate Acclaim C8 120 0.0 Triclosan intoothpaste Reproducible manufacturingpracticesforreproduciblecolumn-to-columnperformance compatible LC-MS Excellent columnef ciencies Low silanolactivityforexcellentpeakshapesbasicanalytes Available in2.2,3and5μmparticle size Triclosan SeparationinToothpasteUsinganAcclaimRSLCC8 Column 1.0 1 200 Minutes 281.1 nm 2.0 400 2 27999 3.0 ek: 1. Saccharin Peaks: triclosan 7.5mg/ of 0.3% +1.0mL containing Toothpaste 1.0g Toothpaste Sonicate and lter. 25mL. make to +methanol sulfate magnesium mL Preparation: 281nm, detector, array Diode Samples: 1.0µL Detection: 0.200mL/min Temperature: 50°C 5 pH acetate Ammonium 2mM Volume: Inj. 15% buffer, Rate: Flow Isocratic, RSLC 3000 Phase: Mobile UltiMate 2.1 ×100mm Buffer: 2.2µm C8, RSLC Acclaim System: HPLC Dimension: Column: Peaks: Preparation: 50°C Samples: Detection: Isocratic,15%buffer, 85%methanol(v/v) Temperature: Inj. Volume: Flow Rate: Mobile Phase: Buffer: PCSse: UltiMate HPLC System: Dimension: 2.1 Column: 2. Triclosan 200–400 nm spectra and time 0.1 sresp. 10Hz, (v/v) methanol 85% 1.0 gToothpaste+mLof7.5mg/mL 1.0 µL 2 mMAmmoniumacetatepH5 2. Triclosan 1. Saccharin 25 mL.Sonicateandlter. magnesium sulfate+methanoltomake Toothpaste containing0.3%triclosan 0.1 sresp.timeandspectra200–400nm Diode arraydetector,281nm,10Hz, 0.200 mL/min Acclaim × 100mm ® ® RSLCC8,2.2µm 3000RSLC Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 2.2μm, 3μm,5μm 2 to 8 2 to 120 11 USP High 300 L7 25 14 LC Columns and Accessories Acclaim Phenyl-1 Hydrophobicity

A unique reversed-phase column with high aromatic selectivity Low High pH Range

0 2 to 8 14 • High aromatic selectivity Pore Size • High hydrophobic retention • Unique and complementary selectivity compared to any other phenyl type column 0 120 300 Carbon Load (%) • Compatibility with highly aqueous mobile phase • High ef ciency and rugged packing 0 13 25 Particle Size 3µm, 5µm USP L11

Acclaim Phenyl-1 columns provide unique selectivity of aromatic compounds for superior chromatographic performance. This column has a higher π-π interaction than other phenyl phases and provides unique selectivity for aromatic compounds while maintaining sufficient hydrophobic interaction and aqueous compatibility

for superior chromatographic performance.

The Acclaim Phenyl-1 column can be used in a wide range of applications in pharmaceutical, environmental, >>

food testing and product-quality testing. This column is ideally suited for the analysis of aromatic Columns for Analytical LC analytes; some examples include glucocorticosteroids, estrogens, fat-soluble vitamins and phospholipids.

Separation of fat-soluble vitamins

60 Column: Acclaim Phenyl-1, 3μm 2 Dimension: 3 × 150mm 3 Mobile Phase: Methanol/water v/v 90/10 Temperature: 30°C Flow Rate: 0.5mL/min Inj. Volume: 2μL Detection: UV at 220nm 4 5 Peaks: (100 ppm each) mAU 6 9 8 1. Retinol acetate (vitamin A acetate) 2. Vitamin D2 3. Vitamin D3 1 10 7 4. delta-Tocopherol 5. gamma-Tocopherol 6. alpha-Tocopherol (vitamin E) 0 7. Impurity (unknown) 8. Vitamin E acetate 0 6 12 18 24 30 9. Vitamin K2 Minutes 27628 10. Vitamin K1

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0 pA 10 Omega fattyacids columns (e.g.C18). structural relatedisomersanduniqueselectivitycomplementarytootherreversed-phase The AcclaimC30isdesignedtoprovidehighshapeselectivityforseparatinghydrophobic • • • • Columns forseparatingstructurallyrelatedisomers Acclaim C30 B A High-quality: lowcolumnbleed,highef ciencyand ruggedpacking Compatibility withhighlyaqueousmobilephase Unique selectivitycomplementarytootherreversed-phasecolumns High shapeselectivity 6000 1 12

0 ng 0 LLA 2 Area 14 3 4 9 5 16 6 Minutes 7 8 9 10 18 11 12 13 20 22 14 28381 24 1. SDA isopropanol in Standards A. 15°C, high lter nebulizer ultra, Corona Peaks: 2µL Samples: Detection: 1.00mL/min 30°C Volume: Inj. Temperature: Rate: Flow %B %A (min) Time Gradient: 675:225:100:4(v/v) Gradient Dual B RS, phase 3000 acid:mobile UltiMate Water:formic A. Phases: Mobile System: LC ×150mm) (4.6 5µm C30, Acclaim Column: UltiMate3000RS,DualGradient AcclaimC30,5µm(4.6×150mm) A.Water:formic acid:mobilephaseB Mobile Phases: LC System: Column: 1.SDA Peaks: A.Standardsinisopropanol Coronaultra,nebulizer15°C, lterhigh Samples: 1000 Detection: 2µL 32 1000 Inj. Volume: 29 30°C 1.00mL/min Temperature: Flow Rate: %B %A Time (min) Gradient: 675:225:100:4(v/v) 7. LLA 6. Arach. 5. DHA 4. GLA 3. ALA 2. EPA B. Saponi edchickenfat B. Acetone:acetonitrile:THF:formicacid 900:3.6:100 (v/v) 7. LLA 7. 6. Arach. DHA 5. GLA 4. ALA 3. EPA 2. B. Saponi ed chicken fat 0 100 0 100 32 595 29 595 24 70 30 22 60 40 0 100 13 1 0 B. Acetone:acetonitrile:THF:formic acid (v/v) 900:3.6:100 4 595 595 24 3070 22 13 4060 1 1000 0 14. Erucic 13. EDA 12. Oleic 11. Adrenic 10. ETA 9.9E,14Z-CLA 8.DPA Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 3µm, 5µm 2 to 8 2 to 13 200 USP L62 High 300 25 14 Acclaim PolarAdvantage Hydrophobicity LC Columns and Accessories Novel polar-embedded reversed-phase columns with unique selectivity Low High pH Range

0 2 to 8 14 • Selectivity complementary to the C18 column Pore Size • Low silanol activity for excellent peak shape with basic compounds 0 120 300 • Compatible with 100% aqueous mobile phase Carbon Load (%) • High selectivity for hydrophobic aromatic molecules 0 16 25 • Wide range of applications Particle Size 2.2μm, 3μm, 5μm USP L60 • Available in 2.2, 3 and 5μm

Acclaim PolarAdvantage (PA) columns feature a patented bonding column chemistry that incorporates a polar sulfonamide group with an ether linkage near the silica surface. This unique chemistry provides low silanol activity, compatibility with 100% aqueous mobile phase. The Acclaim PA column offers great

separation power to resolve a wide variety of polar and nonpolar analytes and supports LC/MS analysis.

Acclaim PA columns provide unique selectivity, good peak shape for acidic, basic, and neutral analytes, >>

and full compatibility with 100% aqueous conditions. Applications include pharmaceutical, environmental, Columns for Analytical LC life science, food testing, and product-quality testing.

EPA 604 phenols

60 Column: Acclaim RSLC PolarAdvantage, 2.2μm 7 Dimensions: 3 × 50mm System: UltiMate 3000 RSLC Mobile Phases: A: 10mM formic acid + 10mM ammonium formate, pH 3.75 ± 0.05 5 B: Acetonitrile Gradient Time (min): -1.5 0.0 0.3 2.6 3.0 3 %A 70 70 70 10 10 2 %B 30 30 30 90 90

mAU Flow Rate: 1.25mL/min Temperature: 30°C 6 Injection Volume: 0.5μL 4 1 8 9 Detection: UV at 280nm, 10Hz, 0.5s resp. time 10 Sample: Calibration mix, 50μg/mL in water Peaks: 1. Phenol 2. 2,4-Dinitrophenol 11 3. 4-Nitrophenol 0 4. 2-Chlorophenol 5. 2-Nitrophenol 0 0.5 1.0 1.5 2.0 2.5 3.0 6. 2,4-Dimethylphenol Minutes 27652 7. 4,6-Dinitro-2-methylphenol 8. 4-Chloro-3-methylphenol 9. 2,4-Dichlorophenol 10. 2,4,6-Trichlorophenol 11. Pentachlorophenol

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0 mAU 0 Turmeric for theseparationofbothbasicandacidicanalytes. The AcclaimPA2 columnisspecificallydesignedtowithstandhighpHconditions,makingitagoodchoice and exhibitshighreversed-phase capacity, with selectivity complementarytoconventional C18columns. unique phases from pH1.5-10with100%aqueousmobile chemistrystability hydrolytic enhanced provides attachmentmulti-point and group polar amide-embedded between the ligands and the silica surface. This Acclaim PolarAdvantageII(PA2) columnsfeatureapatentedsurfacechemistrythatincorporatesan • • • • • • Complementary selectivityandenhancedhydrolyticstability Acclaim PolarAdvantage II Available in2.2,3and5μm Wide rangeofapplications Low bleedforMScompatibility Compatible with100%aqueousmobilephase Novel polar-embedded columnchemistryforhydrolyticstability Unique selectivitycomplementarytotheC18column PA2 C18 1 1 Minutes 1, 2,3 2 2 Total area:9.9mAU*Min. 3 Area: 9.7mAU*Min. 3 25402 4 ek: 1. Curcumin Turmeric extract 428nm UV, Peaks: Sample: Detector: C18: 70% B (v/v) Temperature: 30°C 0.41mL/min Flow: Isocratic: H 15mM A: 2.1 ×100mm 2.2μm, Phase: Mobile C18 120 RSLC Acclaim Dimensions: Columns: Acclaim RSLC PA2 RSLC Acclaim 3. Bis-demethoxycurcumin 2. Demethoxycurcumin B(v/v) 80% PA2: B: Methanol 3 PO 4 Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 2.2μm, 3μm,5μm 1.5 –10.5 120 16 USP High L60 300 25 14 Acclaim HILIC-10 Hydrophobicity LC Columns and Accessories Designed with unique selectivity for hydrophilic molecules Low High pH Range

0 2 to 8 14 • Retains highly polar molecules that are not retained by Pore Size reversed-phase chromatography • Unique selectivity, complementary to reversed-phase columns 0 120 300 Particle Size 3µm, 5µm • Hydrolytically stable • Rugged column packing • Broad application range

The Acclaim HILIC-10 column is designed for separating highly hydrophilic molecules by Hydrophilic Interaction Liquid Chromatography (HILIC). This column is based on high-purity spherical porous silica covalently modified with a proprietary hydrophilic layer.

>> Columns for Analytical LC

Glycerides

200 Column: Acclaim HILIC-10, 3μm Dimensions: 3 × 150mm Mobile Phase: A: Heptane B: 2-Propanol/acetic acid 99.5:0.5 4 Step Elution (min): –6.0 0.0 0.5 4.0 10.0 2 %A 99 99 96 87 87 1 3 %B 1 1 4 13 13 Flow Rate: 0.50mL/min pA 7 Temperature: 25°C 8 Inj. Volume: 4μL 5 Detector: Corona ultra, nebulizer 15°C Peaks: 1. Tristearin 2. Trilaurin 3. Distearin isomer 1 6 4. Dilaurin isomer 1 0 5. Distearin isomer 2 6. Dilaurin isomer 2 7. Monostearin 0 2 4 6 8 10 8. Monolaurin Minutes 27129

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AU 0 Cytosine andnaphthalene (PEGs), ethoxylatedsurfactants,andmore. including non-ionicethoxylatedsurfactants,drugmetabolites,lipids,polyethylene glycols be impossibleforaC18column.Thiscolumnissuitablebroadrangeofapplications, the adjustableselectivity, makingAcclaimMixed-ModeHILIC-1separatemixtures thatwould the terminaldiolgroupfacilitateshydrophilicinteractions.Thisuniquecombination resultsin a diolgroupattheterminus.Thehydrophobicmoietyprovidesreversed-phaseretention and The AcclaimMixed-ModeHILIC-1stationaryphaseconsistsofahydrophobicalkyl chainwith hydrophobic interactionandhydrophilictobeutilizedoptimizeseparations. interaction liquidchromatography(HILIC)properties.Thiscombinationallowsboth HPLC mixed-modestationaryphasethatcombinesbothreversed-phase(RP)andhydrophilic The AcclaimMixed-ModeHILIC-1columnfeaturesaunique,high-efficiency, silica-based • • • • • Uniquely designedforbothreversed-phaseandHILICoperations Acclaim HILIC-1 Mixed-Mode High-ef ciencycolumnforhigh-resolutionseparations Broader applicationrangecomparedwithconventionaldiol-basedcolumns Unique selectivitycomplementarytoRPcolumns Retains highlypolarmolecules Can operateinbothRPandHILICmodes 1 2 2 Reversed-Phase Mode HILIC Mode 4 Minutes 1 2 6 8 24254 10 oiePae CH ×150mm 4.6 Phase: Mobile Dimensions: 5μm HILIC-1, Mixed-Mode Acclaim Column: Peaks: 1. Cytosine (100 ppm) (100 1. Cytosine 254nm at UV 10μL Peaks: 1mL/min Detection: Volume: Inj. Rate: Flow Temperature: 30°C 2. Naphthalene (100 ppm) (100 Naphthalene 2. mode HILIC for 92/8 v/v mode RP for 52/48 v/v 3 CN/0.1 MNH CN/0.1 4 OAc, pH 5.2 pH OAc, Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 3µm, 5µm 2.5 to 7.5 to 2.5 120 High 300 14 Acclaim Mixed-Mode WAX-1 pH Range LC Columns and Accessories Designed for separating anionic molecules; with powerful adjustable 0 2.5 to 7.5 14 selectivity control Pore Size

0 120 300 Particle Size 3µm, 5µm USP L78 • Adjustable selectivity • Selectivity orthogonal to reversed-phase (RP) columns • Ideal selectivity for anionic molecules • Excellent column ef ciency and peak asymmetry • Multimode retention mechanisms: reversed-phase, weak anion exchange, and HILIC modes

The Acclaim Mixed-Mode WAX-1 is a novel, high-efficiency silica HPLC column that combines hydrophobic and weak anion exchange characteristics. Its unique chemistry results in a multimode separation mechanism that includes reversed-phase, anion exchange, and HILIC interactions. Selectivity can be adjusted by changing ionic strength, pH, or organic solvent content.

The Acclaim Mixed-Mode WAX-1 surface consists of a hydrophobic alkyl chain with a tertiary amine group >>

at the terminus. The hydrophobic moiety provides reversed-phase retention and the terminal amino group Columns for Analytical LC facilitates electrostatic interactions.

Pain relief medicine

Column: Acclaim Mixed-Mode WAX-1, 5μm Dimensions: 4.6 × 150mm 3 Mobile Phase: 40/60 v/v Acetonitrile/buffer (6.8 g potassium monophosphate and

0.5 g pyrophosphate in 1000 g D.I. H2O, pH is adjusted to 6.0 with NaOH) Temperature: 30°C 2 Flow Rate: 1mL/min Inj. Volume: 1μL AU 1 Detection: UV, 220nm 4 Peaks: 1. Caffeine 2. Acetaminophen O OH 3. Salicylamide OH 4. Acetyl salicylic acid (Aspirin) 5. Salicylic acid 5

0 10 20 Minutes

www.thermoscienti c.com/chromatography 4-069 4 -070 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories AU 0 Pharmaceutical counterions shapes andexcellentefficiency. molecules (fromhighlyhydrophilictohydrophobic),butalsoseparatesthem withsymmetricalpeak basic retains only not WCX-1 Mixed-Mode Acclaim The products, foodsandbeverages,more. consumer Basic compoundsareimportantinavariety of industrial applications, including pharmaceutical,chemical, normal-phase/ HILIC. and exchange, cation separation modes: reversed-phase, mobile phaseionicstrength,pHororganicmodifier. Thiscolumnthereforecanseparateusingmultiple Selectivity ofionizableandneutralcompoundscanbecontrolledindependentlyorsimultaneouslybytuning a speciallydesignedproprietaryligandwithbothhydrophobicandweakcationexchangeproperties. The AcclaimMixed-ModeWCX-1isanovel,high-efficiency, silica-basedcolumn,manufacturedbybonding • • • • selectivity control Designed forseparatingcationicmoleculeswithadjustable Acclaim WCX-1 Mixed-Mode Multimode separationmechanism:reversed-phase, weakcationexchange,anion-exclusionandHILIC Selectivity complementarytoC18RPcolumns Ideal selectivityforseparatingbasicmolecules selectivity Adjustable 1 2 3 4 Minutes 5 4 6 7 8 Mobile Phase: 40/60 v/v CH v/v 40/60 Phase: Mobile ×150mm 4.6 Dimension: 5μm WCX-1, Mixed-Mode Acclaim Column: 1. Maleate 50μg/mL nm) (225 UV 5µL Peaks: 1mL/min Detection: Volume: Inj. Rate: Flow Temperature: 30°C 7. Timolol 250μg/mL 7. Timolol 300μg/mL Oxprenolol 6. 5. Dexamethasone 60μg/mL 4. Hydrocortisone 60μg/mL 30μg/mL Naproxen 3. 2. Ketoprofen 30μg/mL total) mM (20 5.2 pH 3 CN/NH 4 OAc, Particle Size Pore Size pH Range 0 0 3µm, 5µm 2.5 to 7.5 to 2.5 120 300 14 Acclaim Ordering Guide LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) 120 C18 120 C8 PolarAdvantage PolarAdvantage II 2.2 RSLC Column 30 2.1 071400 072614 072621 071402 3.0 071606 072618 072625 071609 50 2.1 068981 072615 072622 068989 3.0 071605 072619 072626 071608 75 3.0 075697 075696 075698 075699 100 2.1 068982 072616 072623 068990 3.0 071604 072620 072627 071607 150 2.1 071399 072617 072624 071401 250 2.1 074812 074811 074813 074814 3 HPLC Column 33 3.0 066272 – 066274 066276 50 2.1 059128 059122 063174 077999 3.0 068971 – 068972 068973

4.6 059131 059125 – 063189

75 3.0 066273 – 066275 066277 >>

100 2.1 059129 059123 061316 077998 Columns for Analytical LC 3.0 076186 076184 076214 078000 4.6 059132 059126 076216 078001 150 2.1 059130 059124 061317 063187 3.0 063691 068970 063693 063705 4.6 059133 059127 061318 063191 250 2.1 076187 076185 076215 077997 3.0 070077 070078 070079 070080 5 Guard Cartridge 10 2.1 069689 069688 069691 069692 3.0 071981 071979 071983 071985 4.6 069695 069696 069698 069699 HPLC Column 50 2.1 059142 059134 – – 4.6 059146 059138 061319 – 100 2.1 059143 059135 – – 4.6 059147 059139 – – 150 2.1 059144 059136 – – 4.6 059148 059140 061320 063197 250 2.1 059145 059137 – – 4.6 059149 059141 061321 063199

Format Cat. No. Acclaim Guard Cartridge Holder 069580 Acclaim Guard Cartridge-Column Coupler 074188 Acclaim Guard Kit (Holder and Coupler) 069707

www.thermoscienti c.com/chromatography 4-071 4 -072 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Acclaim Ordering Guide 5 5 5 5 3 3 3 3 Particle Size (µm) Size Particle Acclaim Guide Ordering Mixed-Mode (µm) Size Particle Guide Ordering HILIC-10 Acclaim (µm) Size Particle Guide Ordering C30 Acclaim (µm) Size Particle Guide Ordering Phenyl-1 Acclaim HPLC Column HPLC HPLC Column HPLC Column HPLC Guard Cartridge Guard Cartridge Guard Guard Cartridge Guard HPLC Column HPLC HPLC Column HPLC Column HPLC Column HPLC Cartridge Guard Format Format Format Format continued 250 150 250 250 150 150 10 10 250 250 150 150 150 10 100 100 50 50 150 50 10 Length (mm) ID (mm) ID (mm) Length (mm) ID (mm) Length (mm) ID (mm) Length (mm) ID (mm) Length 4.6 4.6 2.1 4.6 4.6 4.6 3.0 4.6 4.6 2.1 2.1 4.6 3.0 2.1 3.0 2.1 3.0 3.0 2.1 4.6 3.0 2.1 2.1 4.6 3.0 2.1 4.6 3.0 2.1 4.6 3.0 2.1 4.6 2.1 4.6 3.0 2.1 3.0 4.6 3.0 2.1 4.6 3.0 2.1 4.6 3.0 2.1 066844 066843 066847 069706 075718 079697 071913 075719 088016 079698 069694 075720 075721 075722 074694 078014 070090 075726 078664 071969 071970 071971 070091 075723 075724 075725 074262 074261 074263 078017 074693 078015 078660 078665 078018 071972 078016 071912 074257 074258 074259 078661 078663 078666 071973 071974 079934 HILIC-1 HILIC-10 C30 Phenyl-1 064985 064984 067084 069704 071909 069686 070088 070089 071908 WAX-1 068352 068353 068371 069705 071911 085455 070092 070093 071910 WCX-1 Notes LC Columns and Accessories

>> Columns for Analytical LC

www.thermoscienti c.com/chromatography 4-073 4 -074 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • of SyncronisHPLCcolumns–columnaftercolumn. Extensive testingandstrongqualitycontrolproceduresensurethe consistency Consistent ReproducibleSeparations,ColumnafterColumn,Time afterTime. Thermo Scienti c Syncronis HPLCColumns Syncronis columnsarebasedonhighlypure Thermo Scienti c 100Å silica,withasurfaceareaof320m speci cations. production ifitmeetsthestringenttest the silicasupportandonlyreleasedfor silica istestedforthephysicalpropertiesof ISO9000 accreditedfacilities.Eachlotof are manufactured,packedandtestedin hydrophobicity, awayfromthesolventfront. good retentionofanalyteshavingarange material. Thisgreatersurfaceareaensures compared to200m ™ and applicationnotes productView information Syncronis 2 /g fortypicalsilicabased ™ HPLCcolumns 2 /g, • • • Syncronis reversedphasecolumnsare Available inthreeparticlesizes: 1.7μmfor available tointeractwithbasicanalytes. minimize thenumberofresidualsilanols densely bondedanddoubleendcappedto for themoretraditionalHPLCanalysis. rapid UHPLCseparationsplus3µmand5μm based onthosedevelopedbyTanaka a seriesofdiagnosticchromatographictests, packed intoSyncroniscolumnsisputthrough Each batchofchromatographicmedia ensure consistent,predictableseparations. www.thermoscienti c.com/syncronis of ourSyncronistechnicalguide range, pleaserequestorviewacopy For fulldetailsontheSyncroniscolumn 1 to • Enhanced, automatedpackingmethodsdrive the requiredquality. individually testedtoensurethatitmeets is consistency evenfurtherandeverycolumn and chelators. and secondaryinteractionswithbases,acids measuring hydrophobicity, shapeselectivity between analytesandthestationaryphase, These testsrigorouslyprobeinteractions LC Columns and Accessories Syncronis C18 Hydrophobicity Syncronis C18 columns deliver consistent predictable separations, Low High pH Range column after column, time after time

0 2 to 9 14 Pore Size • Highly pure, high surface area silica • High carbon load for increased retention 0 100 300 Carbon Load (%) • Double endcapped for extra surface coverage • Highly inert towards basic compounds 0 16 25 • Rigorously tested to ensure quality Particle Size 1.7μm, 3μm, 5μm USP L1

When developing a new method, one of the most important goals for the chromatographer is to achieve a consistent, reproducible separation. The selection of a highly reproducible HPLC column is essential to attaining this goal.

Syncronis C18 columns show excellent column to column reproducibility, >>

as illustrated here by the analysis of zidovudine using five separate columns. Columns for Analytical LC The reproducibility in terms of retention time and peak area is less than or equal to 0.5%, column to column.

Ziovudine

2400 Column: Syncronis C18, 5μm, 150mm x 4.6mm 2000 Mobile phase: Water:Methanol (4:1) Flow rate: 1.0mL min-1 1600 Temperature: 25°C Detection: 265nm 1200

mAU Injection volume: 10μL 800 1. Zidovudine

400

0 0 4 8 12 16 20 24 28 32 Minutes

www.thermoscienti c.com/chromatography 4-075 4 -076 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Uron herbicides • • • • optimum retention hydrophobicity orwhenalesshydrophobicphaseisrequiredtoobtain quicker fromthecolumn.Recommendedforanalyteswithmedium Reduces hydrophobicinteractionsallowingcompoundstoelute C8 Syncronis 1000 mAU Rigorously testedtoensurequality Double endcappedforextrasurfacecoverage Less hydrophobicthanSyncronisC18 Highly pure,highsurfaceareasilica 200 400 600 800 0 0 2 4 1 2 3 6 4 Minutes 5 8 10 6 12 14 16 20μL 1.0mL/min Injection volume.: 10 minutes Bin 60% to 35 Detection: 240nm Temperature: 30°C rate: Flow Gradient: Water A: phase: Mobile x4.6mm 150mm 5μm, C8, Syncronis Column: 6. Linuron Diuron 5. 4. Chlorotoluron 3. Monuron 2. Metoxuron 1. Tebuthiuron Acetonitrile B: Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 1.7µm, 3µm,5µm 100 2 to8 11 USP High 300 L7 25 14 LC Columns and Accessories Syncronis aQ Hydrophobicity

Polar endcapped Syncronis aQ columns provide a controlled Low High interaction mechanism that retains and resolves polar analytes. pH Range

Stable in 100% aqueous mobile phase 0 2 to 8 14 Pore Size

• Stable in 100% aqueous mobile phase 0 100 300 Carbon Load (%) • Enhanced retention of polar compounds

• Rigorously tested to ensure quality 0 19 25 Particle Size 1.7μm, 3μm, 5μm USP L1

In comparison to a conventionally endcapped C18, the Syncronis aQ polar end-capped C18 stationary phase exhibits superior stability towards aqueous mobile phase. Syncronis aQ shows no degradation in performance after 100 injections in a buffered 100% aqueous eluent.

DAD1 A, Sig=260,4 Ref=360,100 (110809B\R2000001.D)

Stability of Syncronis aQ in 100% aqueous mobile phase >>

80 Columns for Analytical LC Column: Syncronis aQ, 5μm, 100mm x 4.6mm

Mobile phase: 50mM Aqueous K2HPO4 (pH 6) 60 Flow rate: 0.7mL/min Temperature: 30°C Detection: 260nm 40 Injection volume: 2μL mAU 1. Cytidine-5’-diphosphate 2. Adenosine-5’-triphosphate 20 3. Adenosine-5’-diphosphate 4. Adenosine-5’-monophosphate 0

0DAD1 A, Sig=260,41 Ref=360,1002 (110809B\R2000085.D)3 4 5 6 7 8 Minutes 100% aqueous inj2 80

60

40 mAU

20

0

0 1 2 3 4 5 6 7 8 Minutes 100% aqueous inj100

www.thermoscienti c.com/chromatography 4-077 4 -078 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • for retentionofaromaticcompounds Provides analternativetoSyncronisC18andisparticularlyuseful Phenyl Syncronis Rigorously testedtoensurequality Highly inerttowardsbasiccompounds Double endcappedforextrasurfacecoverage Alternative selectivitytoSyncronisC18

mAU 100 120 140 20 40 60 80 0 0 Oxacillin sodium(USP) 1 2 3 4 5 6 Minutes 7 8 9 10 11 12 13 14 15 1. Oxacillin Sodium (0.11mg/mL) Sodium 1. Oxacillin Injection volume: 10μL Detection: 225nm Temperature: 25°C method) USP in (2mL/min 1.0mL/min rate: Flow (70:30:10) Mobile phase: Phosphate Buffer: MeCN:MeOH x4.0mm 300mm 5μm, Phenyl, Syncronis Column: Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 1.7µm, 3µm,5µm 100 2 to8 11 USP High L11 300 25 14 LC Columns and Accessories Syncronis Amino Hydrophobicity Provides a versatile aminopropyl phase that gives excellent Low High chromatographic properties in four modes: weak anion exchange, pH Range

reversed phase, normal phase and HILIC 0 2 to 8 14 Pore Size

• Highly pure, high surface area silica 0 175 300 Carbon Load (%) • Double endcapped for extra surface coverage

• Rigorously tested to ensure quality 0 4 25 Particle Size 1.7μm, 3μm, 5μm USP L8

Lactulose

Column: Syncronis Amino 5μm, 150mm x 4.6mm Mobile phase: Water: MeCN (30:70) 2.00 Flow rate: 1.0mL/min

Temperature: 35°C 1.50 Detection: RI Injection volume: 5μL >> 1. Lactulose

1.00 Columns for Analytical LC uRIU

0.50

0.00 -0.25 0 1 2 3 4 5 6 7 8 9 10 Minutes

www.thermoscienti c.com/chromatography 4-079 4 -080 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • of moderatelypolarorganiccompoundsbynormalphasechromatography Serves asapowerfulandefficienttoolforthechromatography Silica Syncronis Rigorously testedtoensurequality Excellent reproducibilityfornormalphasechromatography Highly pure,highsurfaceareasilica

mAU 10 12 0 2 4 6 8 0 Tocopherols 5 10 Minutes 15 025 20 δ γ -tocopherol β -tocopherol α following: the of ug/ml) (200-1000 standards of Mixture 10µL Sample: volume: 2.0mL/min Injection Detection: 254nm (IPA) propan-2-ol Temperature: +0.2% 40°C Hexane rate: Flow phase: Mobile Part Number: 97005-154630 x4.6mm 150 5µm, Silica Syncronis Column: -tocopherol -tocopherol Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 1.7µm, 3µm,5µm 100 2 to8 USP High 300 L3 14 LC Columns and Accessories Syncronis HILIC Hydrophobicity Provides enhanced retention of polar and hydrophilic analytes Low High pH Range

0 2 to 8 14 • Alternative selectivity to Syncronis C18 Pore Size • Improved sensitivity with MS detection 0 100 300 • No need for ion-pair or derivatisation Carbon Load (%) • Outstanding peak shape and sensitivity • Highly pure, high surface area silica particles 0 5 25 Particle Size 1.7μm, 3μm, 5μm • Neutral (uncharged), highly polar surface

Catecholamines Column: Syncronis HILIC, 5µm, 250 x 4.6mm 500 Part Number: 97505-254630 1 Mobile phase: water : acetonitrile : 200mM ammonium formate (10.5 : 84.5 : 5) 400 Flow rate: 1.0mL/min >> Temperature: 40°C 300

Detection: 280nm Columns for Analytical LC Injection volume: 5µL

mAU 2 200 Sample: 1. catechol 4 2. 5-HIAA 3 3. DOPAC 100 4. serotonin 5 6 5. tyrosine 7 6. dopamine 0 7. L-DOPA 0 5 10 15 20 Minutes

www.thermoscienti c.com/chromatography 4-081 4 -082 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Syncronis Ordering Guide Ordering Syncronis UNIGUARD Guard Cartridge Holder Cartridge Guard UNIGUARD Format 5 3 1.7 (µm) Size Particle HPLC Column HPLC Drop-in Guard (4/pk) Column HPLC UHPLC Column Format 10 (mm) Length 250 150 100 50 150 100 50 100 50 30 10 30 (mm) ID 30 (mm) Length 4.0/4.6 3.0 2.1 (mm) ID 4.0/4.6 3.0 4.6 4.6 3.0 4.6 3.0 3.0 4.6 3.0 4.6 4.0 3.0 4.6 4.0 3.0 4.6 3.0 4.6 3.0 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 70-101925040 70-10197905-014001 97905-013001 97305-014001 97305-013001 97205-014001 97205-013001 97105-014001 97105-013001 97203-154630 97103-154630 97103-104630 97203-103030 97103-103030 97103-054630 97103-053030 97102-103030 97102-054630 97102-053030 70-560925243 70-56097905-254630 97305-254630 97905-254030 97205-254630 97305-254030 97105-254630 97205-254030 97105-254030 97205-253030 97105-253030 97205-154630 97905-153030 97205-154030 97105-154630 97305-153030 97105-154030 97205-153030 97105-153030 97105-104630 97905-054630 97105-103030 97305-054630 97205-054630 97105-054630 97205-053030 97105-053030 97105-252130 97205-152130 97105-152130 97105-102130 97905-052130 97305-052130 97205-052130 97105-052130 97103-152130 97103-102130 97203-052130 97103-052130 97102-102130 97202-052130 97102-052130 70-310925023 70-31097905-032130 97305-032130 97205-032130 97105-032130 97105-012101 97103-032130 97102-032130 C18 850-00 852-00 852-00 No. Cat. 97203-104630 – – 97902-103030 97302-103030 97202-103030 97202-054630 – 70-000935133 97905-103030 97205-104630 97305-103030 97205-103030 – 97205-102130 – – 97902-102130 97302-102130 97202-102130 70-111935020 97905-012101 97305-012101 97205-012101 – – C8 97303-154630 97303-104630 97303-103030 – – 97302-054630 – 97305-253030 97305-154630 97305-154030 97305-104630 97305-053030 – 97305-152130 97305-102130 – – 97303-052130 97302-052130 – – aQ 97903-154630 97903-104630 - – – 97902-054630 – 97905-253030 97905-154630 97905-154030 97905-104630 97905-053030 – 97905-152130 97905-102130 – – 97903-052130 97902-052130 – – Phenyl 97705-254630 97705-254030 97705-253030 – 97705-154630 97705-154030 97705-153030 97705-152130 97705-104630 97705-103030 97705-102130 97705-054630 97505-014001 97705-053030 97505-013001 97705-052130 97005-014001 97505-012101 97705-032130 97005-013001 97705-014001 97005-012101 97705-013001 97705-012101 97703-154630 – 97703-104630 - – – – 97703-052130 – 97702-103030 97702-102130 97702-054630 – 97702-052130 – Amino 70-56097505-254630 97505-254030 97005-254630 97505-253030 97005-254030 97005-253030 – 97005-154630 97505-153030 97505-152130 97005-154030 97505-104630 97005-153030 97005-152130 97505-103030 97505-102130 97005-104630 97005-103030 97505-054630 97005-102130 97505-053030 97005-054630 97505-052130 97005-053030 97505-032130 97005-052130 97005-032130 97503-104630 97003-154630 – 97003-104630 – – – 97503-052130 – 97003-052130 97502-103030 – 97502-102130 97002-103030 97002-102130 97502-052130 97002-054630 – 97002-052130 – Silica – 97505-154630 97505-154030 97503-154630 97503-152130 – – – – – 97502-054630 – – HILIC LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) C18 C8 aQ Phenyl Amino Silica HILIC 1.7 UHPLC Column 30 2.1 97102-032130 – – – – – – 50 2.1 97102-052130 97202-052130 97302-052130 97902-052130 97702-052130 97002-052130 97502-052130 3.0 97102-053030 – – – – – – 4.6 97102-054630 97202-054630 97302-054630 97902-054630 97702-054630 97002-054630 97502-054630 100 2.1 97102-102130 97202-102130 97302-102130 97902-102130 97702-102130 97002-102130 97502-102130 3.0 97102-103030 97202-103030 97302-103030 97902-103030 97702-103030 97002-103030 97502-103030 3 HPLC Column 30 2.1 97103-032130 – – – – – – 50 2.1 97103-052130 97203-052130 97303-052130 97903-052130 97703-052130 97003-052130 97503-052130 3.0 97103-053030 – – – – – – 4.6 97103-054630 – – – – – – 100 2.1 97103-102130 – – – – – – 3.0 97103-103030 97203-103030 97303-103030 - - – – 4.6 97103-104630 97203-104630 97303-104630 97903-104630 97703-104630 97003-104630 97503-104630

150 2.1 97103-152130 – – – – – 97503-152130

4.6 97103-154630 97203-154630 97303-154630 97903-154630 97703-154630 97003-154630 97503-154630 >>

5 Drop-in Guard (4/pk) 10 2.1 97105-012101 97205-012101 97305-012101 97905-012101 97705-012101 97005-012101 97505-012101 Columns for Analytical LC 3.0 97105-013001 97205-013001 97305-013001 97905-013001 97705-013001 97005-013001 97505-013001 4.0/4.6 97105-014001 97205-014001 97305-014001 97905-014001 97705-014001 97005-014001 97505-014001 HPLC Column 30 2.1 97105-032130 97205-032130 97305-032130 97905-032130 97705-032130 97005-032130 97505-032130 50 2.1 97105-052130 97205-052130 97305-052130 97905-052130 97705-052130 97005-052130 97505-052130 3.0 97105-053030 97205-053030 97305-053030 97905-053030 97705-053030 97005-053030 97505-053030 4.6 97105-054630 97205-054630 97305-054630 97905-054630 97705-054630 97005-054630 97505-054630 100 2.1 97105-102130 97205-102130 97305-102130 97905-102130 97705-102130 97005-102130 97505-102130 3.0 97105-103030 97205-103030 97305-103030 97905-103030 97705-103030 97005-103030 97505-103030 4.6 97105-104630 97205-104630 97305-104630 97905-104630 97705-104630 97005-104630 97505-104630 150 2.1 97105-152130 97205-152130 97305-152130 97905-152130 97705-152130 97005-152130 97505-152130 3.0 97105-153030 97205-153030 97305-153030 97905-153030 97705-153030 97005-153030 97505-153030 4.0 97105-154030 97205-154030 97305-154030 97905-154030 97705-154030 97005-154030 97505-154030 4.6 97105-154630 97205-154630 97305-154630 97905-154630 97705-154630 97005-154630 97505-154630 250 2.1 97105-252130 – – – – – – 3.0 97105-253030 97205-253030 97305-253030 97905-253030 97705-253030 97005-253030 97505-253030 4.0 97105-254030 97205-254030 97305-254030 97905-254030 97705-254030 97005-254030 97505-254030 4.6 97105-254630 97205-254630 97305-254630 97905-254630 97705-254630 97005-254630 97505-254630

www.thermoscienti c.com/chromatography 4-083 4 -084 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Hypercarb suited a combinationoftwomechanisms: ) Adsorption: 1) The overallretentiononHypercarb with detectiontechniquessuchasMS. for polaranalytesallowsgreatercompatibility use ofincreasedorganicmodifierconcentration buffering systemsorion-pairreagents,and reversed phaseHPLC.Removalofcomplex of analytesthatcannotbeseparatedbytypical allow thesuccessfulretentionandresolution the solute.Thesespecificinteractionmechanisms upon boththepolarityandplanarity(shape)of The mechanismofinteractionisverydependent Retention andResolution aromatic molecule.ThermoScientific satisfied valence,asinaverylargepolynuclear hexagonally arrangedcarbonatomswitha stationary phasecomposedofflatsheets Porous GraphiticCarbon(PGC)isaunique • • • • 100% porousgraphiticcarbonforextendedseparationcapabilities Thermo Scienti c Hypercarb HPLCColumns LC/MS applications. reversed phaseandnormalHPLC polar and theretentionseparationofhighly retentive properties,allowingfortotalpHstability bonded phasesinbothitsstructureand Idealforhightemperatureapplications pHstablefrom0to14 Separatesstructurallyrelatedsubstances Exceptionalretentionofverypolaranalytes contact spatial arrangement. rigid moleculeswitha3-dimensional than planar molecules will showmoreretention contact with theflatgraphitesurface.More molecular areathatisabletocome in the depends uponthesizeandorientation of above. Thestrengthoftheinteraction Hypercarb surface is showninthediagrams non-planar moleculetothe a and planar the points of contact.Theapproacha relationtothegraphitesurfaceat in groups type andpositioningofthefunctional the on largely interactions withHypercarbcolumnsis species. Hypercarbcolumnsareideally to solve “problem”separations,inboth ™ with thegraphitesurface,andalso dependent onthemolecularareain columns are unlike traditional silica silica columnsareunliketraditional The strengthofanalyte ™ columns is

2) Charge inducedinteractionsofapolar Schematic representationofapointchargeapproachingtheHypercarbsurface Schematic representationofmolecularareaaplanarandnon-planarmoleculeinteractingwiththeHypercarbsurface by polaranalytes. molecule withthegraphitesurface. of theelectrostaticchargepolar mechanism isstrictlyduetotheinteraction pH conditions.Thecharge-induceddipole such asabasiccompoundionizedin acidic with theoverallionicchargeofmolecule, surface. These chargesshouldnotbeconfused attraction betweentheanalyteandgraphite an induceddipoleisformed,increasingthe permanent dipoleapproachesthesurface, and accountsforthe charge-induced dipole,isillustratedabove of graphite:Thesecondmechanism, analyte withthepolarizablesurface Asthepolargroupwitha strong retention exhibited strong retentionexhibited development process. Inmostcases,100mm development for aseparation. length columns or shorteraresufficient columns tobeusedduringthemethod Hypercarb columnsusuallyallowsforshorter The strongmechanismsofinteractionwith Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 3µm, 5µm,7µm 250 High 100 300

14 LC Columns and Accessories Increased Retention of Polar Analytes PS-DVB Hypercarb In typical reversed phase chromatography, Courtesy V. Coquart and the retention of an analyte is directly related M-C. Henion, J. Chrom., 1992 to its hydrophobicity: the more hydrophobic the analyte, the longer its retention. Conversely, as the polarity of the analyte increases, analyte-solvent interactions begin to dominate and retention is reduced. This observation holds true for the majority of reversed phase systems. An exception to this rule is Hypercarb columns, for which retention may in some cases increase as the polarity of the analyte increases, illustrated to the right. This phenomenon is referred to as the “polar retention effect Retention on Hypercarb columns increases as polarity of the analyte increases, which is the opposite of typical reversed on graphite” (PREG). This property makes phase materials such as PS-DVB Hypercarb columns particularly useful for the Extended pH Range with typical silica-based columns. Hypercarb separation of highly polar compounds (with columns offer more choice in buffer selection One of the other key benefits of Hypercarb logP as low as -4) that are normally difficult

columns is the extreme stability of the phase while handling both high temperature and >> to retain and resolve on silica-based alkyl to chemical or physical attack. Due to the high pressure. chain phases. The retention of very polar Columns for Analytical LC unique characteristics of the media, it can solutes on Hypercarb columns can be withstand chemical attack across the entire achieved without ion pair reagents or complex pH range of 0 to 14, allowing applications to mobile phase conditions, as illustrated be run at pH levels that are incompatible in the chromatogram below.

Hypercarb column, 5µm, 100 x 0.32mm Part Number: 35005-100365

Mobile Phase: A: H O + 0.1% formic acid columns 2 B: ACN + 0.1% formic acid Gradient: 0 to 25% B in 15 minutes Flow Rate: 8µL/min Temperature: 25°C Detection: UV at 254nm Analytes: 1. Cytosine 2. Uracil 3. Guanine 4. Adenine 5. Xanthine 6. Thymine Additional retention is achieved for polar compounds using a Hypercarb column compared to a polar endcapped C18. Note also the change in elution order.

columns

1 1 4 2

2 Day 1 at pH 12 Day 93 at pH 12 Hypercarb column, 5µm, 100 x 4.6mm 4 Part Number: 35005-104630 Mobile Phase: MeOH:H O 3 2 Isocratic: 70:30 3 Flow Rate: 0.7mL/min Detection: UV at 254nm 5 Sample: 1. Acetone 5 2. Phenol 3. p-Cresol 4. Anisol 6 6 5. Phenetole 6. 3,5 -Xylenol

PGCpH12

0 10 20 Min 0 10 20 Min

Hypercarb column stability at pH 12: retention and selectivity do not change even after 93 days of storage in 0.1M NaOH/MeOH

www.thermoscienti c.com/chromatography 4-085 4 -086 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories antibiotic Axetil.TheHypercarbcolumn with theresolutionofdiastereomers are verysimilarinstructure chromatographic resolutionofcompoundsthat to thegraphitesurface,allowingfor differentiation ofanalytesisbasedontheirfit Hypercarb columns,asshownabove.The considerable resolvingpowerisobservedwith observed usingatraditionalC18column, between methyleneandmethylgroupsis homologous series.Wherenodiscrimination related analytessuchasisomersand columns candifferentiatebetweenclosely way soluteshapeaffectsretention,Hypercarb By virtueofthenaturesurfaceand Related Compounds Resolution ofStructurally Separation ofgeometricisomersAxetil:comparisonaHypercarbandbondedsilicacolumn ACN:H 35005-104630 Phase: Mobile Number: Part x4.6mm 100 5µm, column, Hypercarb Comparison ofmethylandmethylenegroupselectivityonC18Hypercarbcolumns elution order. originally used,aswellachange in in separationoverthesilica-basedcolumn provides bothasignificantimprovement Detector: UV at 254nm at UV 1mL/min Detector: Rate: Flow 38:20:35:10 Isocratic: 01 20 15 10 5 0 Courtesy ofNormanSmith,Glaxo Group Research,Greenford,1988 1 2 Δ3 Δ2 2’ Hypercarb column 1’ 3 Min Anti Isomers 2 O:MeOH:Dioxan 3’ as shownabove 7022

of the

to resolveandquantifythesecompounds. challenges becauseit: A Hypercarbcolumnovercomesthese • • donotprovidethenecessaryreten detection hydrophobic stationaryphaseswhencombined compounds ischallengingbecausethetypical Reversed phase-LC/MSanalysisofverypolar of PolarCompounds Ideal forReversedPhaseLC/MS with themostsuitablemobilephasesforMS Canbeusedwithhighconcentrations of Retainsandseparatesverypolarcompounds sensitivity oftheanalysis. ionization techniques,improvingthe organic modi ers inthemobilephase, ammonium acetateorformate concentrations ofvolatilebufferssuchas 0.1% formicoraceticacidandlow using “MSfriendly”mobilephasessuchas improves nebulizationinatmospheric C18-silica Acid Free Detector: UV at 254nm at UV 1mL/min Detector: Rate: NH Flow MeOH:0.05M 38:62 Isocratic: 30505-204630 Phase: Mobile Number: Part x4.6mm 200 5µm, SAS, Hypersil 015 10 5 0 1 Min Courtesy ofJosef 1’ 2,2’ Kriz etal,1994 pressure pressure 3 which which

4 H 2 PO tion tion 3’ 4

20 • • Hypercarb porousgraphiticcarbon Surface comparisonbetweenC18bondedsilicaand Typical C18silica Hypercarb Is stable with any mobile phase and Isstablewithanymobilephaseand Allowsshortercolumnlengthsandsmaller surface isnotmodi ed. the Hypercarbcolumn’s porousgraphitic produces nophasebleedissuesbecause MS techniques. capillary columnsaremorecompatiblewith The  owratesusedwithnarrowboreand peak capacity, oftenwithincreasedsensitivity. diameters tobeusedwithoutcompromising Hypercarb Ordering Guide LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 3 Drop-in Guard (4/pk) 10 1.0 35003-011001 2.1 35003-012101 3.0 35003-013001 4.6 35003-014001 HPLC Column 30 1.0 35003-031030 2.1 35003-032130 3.0 35003-033030 50 1.0 35003-051030 2.1 35003-052130 3.0 35003-053030 4.6 35003-054630 100 1.0 35003-101030 2.1 35003-102130 3.0 35003-103030 4.6 35003-104630 150 2.1 35003-152130

3.0 35003-153030

4.6 35003-154630 >> High Temperature HPLC Column 30 2.1 35003-032146 50 2.1 35003-052146 Columns for Analytical LC 4.6 35003-054646 100 1.0 35003-101046 2.1 35003-102146 3.0 35003-103046 4.6 35003-104646 5 Drop-in Guard (4/pk) 10 1.0 35005-011001 2.1 35005-012101 3.0 35005-013001 4.6 35005-014001 HPLC Column 30 2.1 35005-032130 3.0 35005-033030 4.6 35005-034630 50 1.0 35005-051030 2.1 35005-052130 3.0 35005-053030 4.6 35005-054630 100 1.0 35005-101030 2.1 35005-102130 3.0 35005-103030 4.6 35005-104630 150 1.0 35005-151030 2.1 35005-152130 3.0 35005-153030 4.6 35005-154630 High Temperature HPLC Column 30 2.1 35005-032146 4.6 35005-034646 50 2.1 35005-052146 4.6 35005-054646 100 2.1 35005-102146 4.6 35005-104646 Javelin HTS Column 20 2.1 35005-022135 Preparative HPLC Column 100 10 35005-109070 21.2 35005-109270 30 35005-109370 150 10 35005-159070 21.2 35005-159270

Format Length (mm) ID (mm) Cat. No. UNIGUARD Guard Cartridge Holder 10 1.0 851-00 2.1 852-00 3.0 852-00 4.6 850-00

www.thermoscienti c.com/chromatography 4-087 4 -088 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 5μm, 300ÅforAcclaimSEC-300and7μm,1000SEC-1000. and oligomers.AcclaimSECcolumnsareavailableintwosizestocoverabroad molecularweightrange: exclusion chromatographycolumnsspecificallydesignedfortheseparationofwater solublepolymers Thermo Scientific • • • • • High PerformanceSECColumnsforAnalysisofWater SolublePolymers Acclaim Size Exclusion (SEC) Chromatography

Molecular Weight (Da) Stable surfacebondingwithlowcolumnbleedandcompatibilityUV, RI,MS,ELSDand Availability ofsmallparticlesizes packedin4.6x300mmdimensionallowsfor high-resolutionanalysis SEC-1000 calibratedfrom1,000to1,000,000Daltons SEC-300 calibratedfrom100to50,000Daltons Proprietary mono-dispersedmulti-porehydrophilicresin:noin ectionpointsincalibrationcurve Thermo Scienti c at reducedsolventconsumption 100,000,000 10,000,000 1,000,000 100,000 10,000 1000 100 10 1.50 ™ Acclaim 2.00 ™ Dionex 2.50 ™ Elution Volume(mL) ™ SEC-300andSEC-1000areafamilyofresinbased,high performancesize Corona 3.00 ™ ChargedAerosolDetectors 3.50 SEC-300 SEC-1000 .04.50 4.00

oun: Acam SEC-300 Acclaim Columns: ape: phase) -0.1% mobile in (0.03% RI Samples: 0.35mL/min Detection: 50μL Injection Volume: Temperature: 25°C perchlorate sodium 10mM x300mm 4.6 rate: Flow Phases: Mobile Dimensions: AcclaimSEC-1000 and ethylene glycol (MW 62) 62) (MW glycol ethylene and 106 (MW glycol diethylene 200), and 400 600, 1,500, 3,000, 8,300, (MW PEG 20,000), and 43,000, 580,000, 272,000, 185,000, 80,000, 40,000,000), PEO (MW 895,000, dextran (MW 5,000,000- Polyacrylic acid using size-exclusion chromatography with charged-aerosol detection LC Columns and Accessories

2.5 HPLC Conditions pA Column: Acclaim SEC-300 PAA Dimensions: 5µm, 4.6 x 300mm System: UltiMate 3000 RS Mobile Phases: A: Acetonitrile 400 µg/L B: Water Isocratic: 10:90 200 µg/L Flow rate: 0.35mL/min 100 µg/L Injection: 35µL Temperature: 30°C 50 µg/L Detection: Corona III; evaporator 55°C, Engine 40 °C, 2 Hz, lter 5, power function 25 µg/L -0.0 1.20 Peaks: 1. PAA standards in water Polyacrylic acid 0.450 0 5 10 15 Minutes Area [ pA *min]

0.200

Concentration [ µg/L]

0.00c0 0 125 250 >> Columns for Analytical LC

Acclaim SEC Ordering Guide Description Particle Size (µm) Format Length (mm) 4.6 mm ID 7.8 mm ID Acclaim SEC-300 5 Guard 33 082740 – HPLC Column 150 – 079726 300 079723 079725 Acclaim SEC-1000 7 Guard 33 082739 – HPLC Column 150 – 079722 300 079724 079721

www.thermoscienti c.com/chromatography 4-089 4 -090 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 8330, withcomplementaryselectivity. separation ofall14explosivesinEPA Method Optimized columnchemistryforbaseline Acclaim Explosives classes ofsurfactants. column specificallyfortheseparationofall The mostversatilecommercially-available Acclaim SurfactantandPlus aliphatic, andaromaticorganicacids. Designed forseparationofhydrophilic, Acclaim OrganicAcid neutral drugs. as wellmixturesofacidic,basic,and drug substancesandcounterionanalysis, simultaneous separationofpharmaceutical Application specificcolumnsdesignedforthe Acclaim Trinity P1andP2 provide superiorresolutionwithease-of-use. Thermo Scientificapplicationspecificcolumnsarebasedonnovelanduniquechemistries and • • • • • • • • Innovative chemistriestailoredforchallengingandcriticallyimportant applications Thermo Scienti c ApplicationSpeci c LC Columns Hypersil GreenPAH forpolyaromatichydrocarbonanalysis Acclaim CarbonylC18foraldehydeandketoneseparation Acclaim Carbamatefortheseparationofcarbamateinsecticides Acclaim Trinity Q1fordiquatandparaquatanalysis Acclaim Explosivesforseparationofexplosiveresidues Acclaim SurfactantandPluscolumnsforseparationofsurfactants Acclaim OrganicAcidforfastorganicacidanalysis Acclaim Trinity P1andP2forAPIcounterionanalysis specified inEPA Method610. analysis ofpolyaromatichydrocarbons(PAHs), carbon loading,designedspecificallyforthe Specially tailoredalkylbondedsilicawithahigh Hypersil GreenPAH EPA Method8315andrelatedmethods. using CARBMethod1004,EPA Method554, andketonesinair, waterandsoil aldehydes of Designed forseparatingDNPHderivatives Acclaim CarbonylC18 Method 531.2. carbamate pesticidespecifiedinUSEPA A specificallycolumnfortheseparationof Acclaim Carbamate diquat andparaquat. designed fortheseparationofherbicides Unique, high-efficiency, silica-basedcolumns Acclaim Trinity Q1 Acclaim Trinity P1 LC Columns and Accessories Most innovative advancement in mixed mode column technology: reverse-phase, anion- and cation exchange functionality on a single support

• Ideal selectivity for simultaneous separation of API and counterion • Adjustable selectivity by mobile phase ionic strength, electrolyte type, pH, and organic solvent • Low bleed; compatible with MS, CAD and ELSD. • Retention of hydrophilic ionic and ionizable analytes without ion-pairing reagents • Greater  exibility in method development: each retention mechanisms can be controlled independently

The Thermo Scientific™ Acclaim™ Trinity™ P1 HPLC column is designed with unique multimode surface chemistry ideal for the simultaneous separation of drugs and their counterions. The surface chemistry concurrently provides reversed-phase, cation exchange, and anion exchange functionalities. The result is maximum flexibility in method development. Separations can be optimized easily by adjusting the chromatographic

parameters (mobile phase pH, ionic strength, and organic strength).

The Acclaim Trinity P1 stationary phase, based on this Nanopolymer Silica Hybrid (NSH) technology, consists of high-purity porous, spherical 3μm silica >>

particles, coated with charged nanopolymer beads. The unique surface chemistry includes an inner-pore area modified with an organic layer that Columns for Analytical LC provides both reversed phase and anion exchange properties. The outer-pore surface, conversely, is modified with cation exchange functionality.

Acclaim Trinity P1 Ordering Guide Particle Size (µm) Format Length (mm) 2.1mm ID 3.0mm ID 3 Guard Cartridges 10 071391 071390 (2/pk) HPLC Column 50 075565 071388 100 071389 071387 150 075564 075563

Acclaim Guard Holder Ordering Guide Format Cat. No. Acclaim SST Guard Cartridge Holder V-2 069580 Acclaim Guard Kit (Holder and coupler) V-2 069707 Guard to Analytical Column Coupler V-2 074188

Simultaneous separation of pharmaceutical counterions

300 Column: Acclaim Trinity P1, 3μm Dimensions: 3.0 × 100mm

Mobile Phase: 60/40 v/v CH3CN/20mM (total)

NH4OAc, pH 5 Temperature: 30°C Flow Rate: 0.5mL/min 3 Inj. Volume: 2μL Detection: Corona ultra (Gain = 100 pA;

mV 1 Filter = med; Neb Temp = 30°C) 4 Peaks: (50 to 100ppm) 2 1. Choline 2. Tromethamine 3. Sodium 8 4. Potassium 5 6 5. Meglumine 7 9 10 6. Mesylate 7. Nitrate 0 8. Chloride 9. Bromide 0 3 6 9 12 15 10. Iodide Minutes

www.thermoscienti c.com/chromatography 4-091 4 -092 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories pharmaceutical counterionanalysisbyHPLC. exchange andcation-exchangeregions.AcclaimTrinity P2columnisaimedtocomplementAcclaimTrinity P1toprovideatotalsolutionfor retention, whiletheoutersurfaceismodifiedwithanion-exchangenano-polymer beads.Thischemistryensuresspatialseparationoftheanion- nanopolymer particles.Theinner-pore areaofthesilicabeadismodifiedwitha covalentlybondedorganiclayerthatprovidescation-exchange The AcclaimTrinity P2columnisbasedonNSHtechnology, whichconsistsofhigh-purityporoussphericalsilicaparticlescoatedwithcharged monovalent anddivalentcationsoranions. The AcclaimTrinity P2isaunique,high-efficiency, silica-basedcolumnspecificallydesignedforseparationofpharmaceuticalcounterions,including • • • • • anion exchangeandcationfunctionalities Most innovativemixed-modecolumntechnology;hydrophilicinteraction, HILIC, Acclaim P2 Trinity 5.0 High ef ciency stable Hydrolytically Low columnbleed,compatiblewithCADandMS Selectivity complementarytotheTrinity P1column Ideal forseparatingpharmaceuticalcounterions,includingmonovalentanddivalent pA 3 Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard Acclaim Format Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering P2 Trinity Acclaim cations oranions 0 Pharmaceutical-related anionsandcations 0 1 2 3 63 4 5 (2/pk) Cartridges Guard Format HPLC Column HPLC 6 7 12 Minutes 8 9 18 10 11

10 Length (mm) Length 100 50 24 12

0 085435 2.1 mm ID 3.0 mm ID mm 3.0 2.1 ID mm 085432 085431 Time (min) Time 1. Phosphate water D.I. in each –0.10 mg/mL 0.02 Detector Aerosol Charged Veo Corona Peaks: mL/min 0.60 Samples: Detection: 2μL Injection volume: rate: Flow pH Temperature: NH4OFm, mM 100 30°C and water D.I. mm x100 3.0 phase: Mobile Dimensions: 3µm P2, Trinity Acclaim Column: 074188 069707 069580 No. Cat. -10 22 30 2 0 0.760 H 80 80 0 0 2 O 12. Calcium 12. 11. Magnesium 10. Sulfate Fumarate 9. 8. Citrate Nitrate 7. 6. Bromide 5. Malate 4. Chloride 3. Potassium 2. Sodium gradient 3.65 0.1 M Ammonium formate, pH3.65 formate, 0.1 MAmmonium 085436 085434 085433 1.474 100 100 20 20 Acclaim Organic Acid LC Columns and Accessories Optimized and application-tested for the analysis of hydrophilic organic acids

• Use-tested to guarantee consistent hydrophilic organic acid separations • Compatible with 100% aqueous mobile phases • Hydrolytic stability at low-pH conditions • Ideal selectivity for separating a wide spectrum of organic acids • Excellent column ef ciency and peak shapes for organic acids

The Acclaim Organic Acid (OA) is a silica-based reversed-phase column designed for high-efficiency, high-throughput organic acids analysis. It offers unparalleled performance for separating hydroxyl aliphatic and aromatic organic acids. The Acclaim OA is the recommended column for determining small hydrophilic organic acids, C1 to C7 aliphatic acids, and hydrophilic aromatic acid and is also valuable for the analysis and quality assurance of food and beverage products, pharmaceutical preparations, plating baths, and manufacturing chemicals, chemical intermediates, and environmental samples.

>> Columns for Analytical LC

Acclaim Organic Acid Ordering Guide Particle Size (µm) Format Length (mm) 2.1mm ID 3.0mm ID 4.0mm ID 3 HPLC Column 150 070087 070086 – 5 Guard Cartridges 10 – 071987 069700 (2/pk) HPLC Column 150 – – 062903 250 – – 062902

Acclaim Guard Holder Ordering Guide Format Cat. No. Acclaim SST Guard Cartridge Holder V-2 069580 Acclaim Guard Kit (Holder and coupler) V-2 069707 Guard to Analytical Column Coupler V-2 074188

Hydrophilic organic acids Column: Acclaim OA, 5μm, 4 × 250mm 0.025 Mobile phase: 100mM Na2SO4, pH 2.65 10 (adjusted with methanesulfonic acid) Temperature: 30°C Flow rate: 0.6mL/min Injection volume: 5μL Detection: UV, 210nm

AU 2 Peaks: 1. Oxalic acid 15mg/L (ppm) 2. Tartaric acid 120 3. Formic acid 180 1 3 4 12 4. Malic acid 120 8 5. iso-Citric acid 120 6 7 5 9 11 6. Lactic acid 180 7. Acetic acid 120 0 8. Citric acid 120 0 5 10 15 9. Succinic acid 120 Minutes 20011 10. Fumaric acid 7 11. cis-Aconitic acid * 12. trans-Aconitic acid * * 7ppm total for cis and trans isomers

www.thermoscienti c.com/chromatography 4-093 4 -094 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories

mV of thesesurfactants. petroleum products,cosmetics,andpharmaceuticals.TheAcclaimSurfactantcolumn wasdesignedspecificallyforHPLCseparation Surfactants arewidelyusedinindustrial,agricultural,andpharmaceuticalmarkets, inproductsasdiversepesticides,detergentspowders, resolution betweenstronglyhydrophiliccompounds,suchasisomersofxylene sulfonate. Acclaim Surfactantcolumnsarealsoresistanttodewettingunderhighlyaqueous mobilephaseconditions,andthuscanbeusedtoprovideexcellent oligomers ofethoxylatedsurfactantscomparedwithconventionalC18columns. As aconsequenceofitsnovelchemistry, thiscolumnexhibitsauniquepolarity that providessignificantlyimprovedresolutionforindividual surfactants, includinganionic,cationic,nonionic,andamphotericsurfactantsusing UV, ELSDorRIdetection. The AcclaimSurfactantcolumnsarethefirstgenerationhigh-efficiency, silica-basedcolumnsdesignedspecificallyforseparatingawidevarietyof • • • • • Excellent performanceforseparatingabroadrangeofsurfactants Acclaim Surfactant 5 0 Inorganic anion,hydrotropes,cationic,nonionic,amphoteric,and anionicsurfactants Rugged separationsunderavarietyofconditions Improved resolutionforethoxylatedsurfactants Compatible withhighlyaqueousmobilephases Excellent peakshapes,especiallyforcationicsurfactants Ideal selectivityforseparationofanionic,nonionic,cationicand Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard SST Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering Surfactant Acclaim 3 5 amphoteric surfactants 1 2 3 4 10 5 6 Format HPLC Column HPLC (2/pk) Cartridges Guard HPLC Column HPLC 7 15 Minutes 8 20 9 10 11 25 Length (mm) Length 150 10 150 250 12 13 14 30 22793 35 074188 069707 069580 No. Cat. 2.1mm ID 2.1mm 070085 069693 068123 – 11. C 11. 10. Dodecyl sulfate sulfate Decyl 9. 8. betaine Cetyl X-100 7. Triton 6. chloride Lauryldimethylbenzyl-ammonium chloride Laurylpyridinium 5. sulfonate Xylene 4. Nitrate 3. 2. Bromide ELS detector 1. Chloride 25µL Peaks: 1mL/min Detection: Volume: Inj. 30°C 25min, Ain 85% to 25% Rate: Flow Temperature: Gradient:

12. C 12. Mobile Phase: (A) CH (A) ×150mm 4.6 5µm Surfactant, Acclaim Phase: Mobile Dimensions: Column: 13. C 13. 14. C 14. 3.0mm ID 070084 071991 – – 10 12 11 13 -LAS -LAS -LAS -LAS then hold 85% A for 10min Afor 85% hold then (B) 0.1 MNH (B) 3 CN, CN, 4 OAc, pH 5.4 pH OAc, 4.6mm ID 4.6mm – 069701 063201 063203 Acclaim Surfactant Plus LC Columns and Accessories Column of Choice for Surfactant Analysis using Higher Sensitivity Detection: Performance, Versatility, Throughput

• Ideal selectivity for simultaneous separation of anionic, nonionic, cationic, and amphoteric surfactants • Compatible with multiple detectors including MS, CAD, ELSD and UV • Well suited for the determination of cationic surfactants • High ef ciency and fast analysis • Rugged separations under a variety of conditions

Acclaim Surfactant Plus is a new generation of columns offering improved performance and higher throughput for analyzing surfactants. They have similar selectivity to the Acclaim Surfactant columns, but have been designed with exceptionally low bleed for use with charged aerosol detectors

(CAD) and mass spectrometers (MS). These columns can be used to separate a wide variety of surfactants including anionic, cationic, nonionic and

amphoteric surfactants. >>

Additionally, these columns can be used with evaporative light scattering detectors (ELSD), suppressed conductivity detectors (SCD), and UV-Vis Columns for Analytical LC detectors (UV). They are resistant to dewetting under highly aqueous mobile phase conditions, and thus can be used to provide excellent resolution between strongly hydrophilic compounds, such as isomers of xylene sulfonate. Non-metallic PEEK hardware is available for best compatibility with Dionex ion chromatography systems.

Cationic surfactants Column: Acclaim Surfactant Plus, 3μm Dimensions: 3.0 × 150 mm 10 1 System: ICS 3000 Mobile phases: A: Acetonitrile µS B: 100mM Formic acid C: Water Gradient: Time 2 3 %A %B %C 4 (min) -12 5 5 90 0 5 5 90 0 12 40 5 55 20 40 5 55

Flow Rate: 0.5 mL/min Injection: 5 μL 6 5 7 Temperature: 25°C 5 Detection: Conductivity with blank subtraction µS 8 Suppressor: CSRS300-2mm (external water 1.0 mL/min, current = 8 mA) Peaks: 1. Tetrabutylammonium 2. Tetrapentylammonium 3. Tetrahexylammonium 0 4. Tetraheptylammonium 5. Decyl-trimethylammonium 6. Dodecyl-trimethylammonium 7. Tetradecyl-trimethylammonium 0 5 10 15 8. Hexadecyl-trimethylammonium Minutes

www.thermoscienti c.com/chromatography 4-095 4 -096 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard SST Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering Plus Surfactant Acclaim 3 5 Format HPLC Column HPLC (2/pk) Cartridges Guard HPLC Column HPLC Length (mm) Length 100 150 250 10 250 150 2.1mm 078955 078954 078953 078960 – – 3.0 mm 078952 078951 – 078959 – – 074188 069707 069580 No. Cat. 4.6 mm 4.6 – 078950 – 082773 082767 082768 4.0 mm PEEK mm 4.0 – – – – – 078956 Acclaim Explosives LC Columns and Accessories The best solution for explosives analysis (EPA Method 8330)

• Both Acclaim E1 and E2 columns provide baseline resolution of all 14 compounds targeted by EPA Method 8330 • E2 columns available in 2.2, 3 and 5μm particle size • The E1 and E2 columns have mutually complementary selectivity • Simple isocratic elution conditions • Rugged columns with good lot-to-lot reproducibility

Acclaim Explosives E1 and E2 columns are specifically designed to resolve all 14 explosives listed in EPA SW-846 Method 8330: Nitroaromatics and Nitramines by HPLC. The novel and unique chemistries of these columns provide superior resolution with complementary selectivities. The Acclaim Explosives E1 is recommended for use as a direct replacement for ODS columns for the primary analysis. The Acclaim Explosives E2

may be used as either a primary or a confirmatory column. The unique selectivity and versatility of Acclaim Explosives E2 column provides

a wider application range, including the analysis of explosives beyond U.S. EPA Method 8330 (ISO22478). >> Columns for Analytical LC

Complimentary baseline separation of 14 target compounds listed in EPA SW-846 method 8330

4 3 Acclaim Explosives E1 Column: 5µm, 4.6 x 250mm 1 Column: 5µm, 4.6 x 250mm 2 5 7 11 Eluent: EPA 8330 mix, 50 ppm each 6 9 8 10 Temperature: 30 °C 12 14 13 Flow Rate: 1 mL/min Inj. Volume: 5µL 4 Detection: UV, 254nm AU 3 Acclaim Explosives E2 Peaks: 1. HMX 2. RDX 5 11 3. 1,3,5-Trinitrobenzene 7 4. 1,3-Dinitrobenzene 2 1 6 10 5. Nitrobenzene 12 9 13 14 8 6. Tetryl 7. 2,4,6-Trinitrotoluene 8. 4-Amino-2,6-Dinitrotoluene 9. 2-Amino-4,6-Dinitrotoluene 0 9 18 27 36 45 0. 2,6-Dinitrotoluene Minutes 22787 11. 2,4-Dinitrotoluene 12. 2-Nitrotoluene 13. 4-Nitrotoluene 14. 3-Nitrotoluene

Rapid determination of EPA 8330A explosives Column: Acclaim RSLC Explosives E2, 2.2µm 60 4 Dimension: 2.1 × 100mm HPLC System: UltiMate 3000 RSLC HPG Mobile Phases: Methanol:water 48:52 (v/v) 3 Flow Rate: 0.34mL/min (293 bar) Injection Vol.: 1µL Temperature: 31°C 9 5 Detection: UV at 254nm, 10 Hz, 0.4 s resp. time 6 Sample: Calibration mix, 25µg/mL

mAU in 50% acetontrile 2 8 Peaks: 1. HMX 8. 2,6-DNT 1 7 14 2. RDX 9. 2,4-DNT 10 12 13 3. 1,3,5-TNB 10. 2-NT 11 4. 3,5-DNB 11. 4-NT 5. NB 12. 3-NT 6. 2,4,6-TNT 13. 4-Am-2,6-DNT 0 7. Tetryl 14. 2-Am-4,6-DNT

0 2.5 5 7.5 10 Minutes 28815

www.thermoscienti c.com/chromatography 4-097 4 -098 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Acclaim Explosives Ordering GuideAcclaim Explosives 5 3 2.2 5 E1 and E2 Guard Cartridges (4.6 x 10mm), pkg of 2 of pkg x10mm), (4.6 Cartridges Guard E2 E1 and x250mm) (4.6 Columns HPLC E2 E1 and Description Guide Ordering Kit Explosives Acclaim V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard SST Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering E2 Explosives Acclaim (µm) Size Particle Guide Ordering E1 Explosives Acclaim HPLC Column HPLC (2/pk) Cartridges Guard HPLC Column HPLC HPLC Column HPLC RSLC Column RSLC (2/pk) Cartridges Guard Format Format 250 10 250 150 250 150 100 10 Length (mm) Length (mm) Length – – – 070083 – 076226 076225 – 2.1mm ID 2.1mm ID 2.1mm – 071989 070081 070082 – – 076227 – 3.0mm ID 3.0mm ID 074188 069707 069580 No. Cat. 064312 No. Cat. 064309 069703 – – 064305 – – 069702 4.6mm ID 4.6mm ID 4.6mm Acclaim Trinity Q1 LC Columns and Accessories For Trace Analysis of Diquat and Paraquat

• Excellent resolution of diquat and paraquat • Good peak shape • Fast analysis • LC/MS compatibility • No ion-pairing reagent needed • Easy to use

Acclaim Trinity Q1 columns are unique, high-efficiency, silica-based columns designed for the separation of the herbicides diquat and paraquat. These herbicides are toxic and residues are monitored in drinking water, wastewater and agricultural products. The Acclaim Trinity Q1 column is a tri-mode (WCX, WAX, RP), column based on Nano-polymer Silica Hybrid technology. It offers unmatched high-resolution and high-throughput trace analysis of the herbicides diquat and paraquat by LC-MS/MS and LC-UV methods.

>> Columns for Analytical LC Diquat and paraquat

Chromatographic Conditions Paraquat: Q-SRM System: 1.19E5 RSLC system Column: Acclaim Trinity Q1, 3µm Column Temp: Ambient 185/169 Mobile Phase: 25% Ammonium Acetate (100 mM, pH 5.0); 75% Acetonitrile Paraquat: C-SRM Flow Rate: 0.5 mL/min 5.46E4 Injection:

Mass Spectrometric Conditions 185/170 System: MAX Quadrupole Mass Spectrometer Interface: Heated Electrospary Ionization Paraquat-IS with HESI II probe 5.47E4 Spray Voltage: 1500 V Vaporizer Temp: 400 ºC Sheath Gas Pressure: 70 193/178 Aux Gas Pressure: 10 Capillary Temp: 350 ºC Quantitation Mode: Selected Reaction Monitoring (SRM) Diquat: Q-SRM 9.67E5 Scan Events Precursor Quantitative SRM (CID) SRM (CID) Relative Abundance 183/157 Paraquat 185 169 (27) 170 (17) Paraquat-d6 193 178 (17) Diquat 183 157 (22) 130 (31) Diquat: C-SRM Diquat-d 186 158 (22) 3.11E5 3

183/130

Diquat-IS 9.98E5

186/158

0.50.0 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Time (min)

Acclaim Trinity Q1 Ordering Guide Particle Size Length (mm) 3.0 mm ID 2.1 mm ID 3 HPLC Column 50 079716 079718 5 Guard Cartridges 10 079719 079720 (2/pk)

www.thermoscienti c.com/chromatography 4-099 4 -100 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories EPA Method 531.2 that uses HPLC with postcolumn derivatization. LC-MS is the method of choice fortheultimatesensitivity. of method the is LC-MS EPA Method531.2thatusesHPLCwithpostcolumnderivatization. Drinking waterandrawsurfaceismonitoredforthepresenceofcarbamatepesticidesrelatedcompoundsusinganestablished specified inUSEPA Method531.2.Carbamatepesticidesarewidelyusedthroughouttheworld. Acclaim Carbamatecolumnsaredesignedforbaselineseparationofcarbamates(N • • • • • specified inUSEPA Method531.2 Designed forbaselineseparationofcarbamatepesticides Acclaim Carbamate counts counts 2000 4000 -500 -300 High-ef ciency, extremelylowcolumnbleed,andruggedpacking Compatible withbothbinary(methanol/water)andternary(acetonitrile/methanol/water) Use witheitherLC/postcolumnderivatization/ uorescence orLC/MSdetection Baseline separationofcarbamatepesticidesspeci edinUSEPA Method531.2 5 3 2.2 Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard SST Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering Carbamate Acclaim Available in2.2,3and5μmparticle size mobile phasegradients 0 0 Carbamate standard-spikedricesamples B: withdispersiveSPEusingPSA A: withoutdispersiveSPE 2 1 1 1.3 2 2.5 HPLC Column HPLC HPLC Column HPLC (2/pk) Cartridges Guard RSLC Column RSLC Format 3 1 1 3.8 2 2 4 3 3 4 4 5 5 250 150 10 150 100 Length (mm) Length 6 6.3 5 5 7 7.5 Minutes 6 6 8 8.8 9 – 072927 072930 075596 075597 2.1mm ID 2.1mm 10 10 8 7 7 8 11 11.3 9 9 10 10 12 12.5 – 072926 072929 – – 3.0mm ID -methylcarbamate and N -methylcarbamate and 074188 069707 069580 No. Cat. 13 13.8 14 11 11 REF: 28175 12 12 15 15 A B 16 16 072924 072925 072928 – – 4.6mm ID 4.6mm oiePae Methanol–H Mobile Phase: 3μm ×150mm, 3.0 Carbamate, Acclaim Column: Peaks: and Excitation/330nm Detection: Fluorescence 2: 1 and Reagents of Rate Flow reaction second regent, OPA 2: Reagent Postcolumn coil NaOH, reaction 0.2% rst 1: 250μL Reagent Postcolumn ºC 50 for each 0.4μg/L component Standard Concentration: Injection Volume: Rate: Flow Temperature: Gradient: In -methylcarbamoyloxime pesticides) 12. BDMC (I.S.) BDMC 12. 11. Methiocarb 10. 1-Naphthol 9. Carbaryl 8. Carbofuran 7. Propoxur 6. Aldicarb carbofuran 3-Hydroxy 5. 4. Methomyl Oxamyl 3. sulfone Aldicarb 2. sulfoxide 1. Aldicarb Emission/465nm 0.3mL/min temperature room at coil ºC 100 at 0.9mL/min 70% min, 13.6–16 40%; min, 8.0 20%; min, 2.0 14%; min, -4.0–0.0 Methanol, 2 O Acclaim Carbonyl C18 LC Columns and Accessories A silica-based, reversed-phase column designed specifically for separating DNPH derivatives of aldehydes and ketones

• Ideal selectivity for baseline resolution of DNPH derivatives of aldehydes and ketones regulated by various of cial methods, including EPA 554, EPA 8315, EPA 1667, EPA TO-11, and CARB 1004 • High ef ciency for UHPLC performance • Rugged columns with good lot-to-lot reproducibility • Proven robust methods

Acclaim Carbonyl C18 columns are silica-based reversed phase columns designed specifically for separating DNPH derivatives of aldehydes and ketones. They exhibit superior resolution compared with other commercially available columns. Aldehydes and ketones are common pollutants in air and water. The analytical difficulties that need to be overcome include their volatility, their reactivity, and their modest UV absorption. The reaction with dinitrophenylhydrazine (DNPH) is a convenient means of trapping, stabilizing, >> and tagging these substances. Several standard methods have been developed to apply this chemistry to various environmental situations. Some of the better known ones include CARB 1004 for vehicle exhaust, EPA 554 for drinking water, EPA 1667 for pharmaceutical wastewater, Columns for Analytical LC and EPA 8315 for general wastewater.

Acclaim Carbonyl C18 Ordering Guide Particle Size (µm) Format Length (mm) 2.1mm ID 3.0mm ID 4.6mm ID 2.2 RSLC Column 100 077972 077974 – 150 077973 – – 3 HPLC Column 150 079011 079010 – 250 – 079009 – 5 Guard Cartridge 10 079012 079013 079014 (2/pk) HPLC Column 150 – – 079008 250 – – 083214

DNPH aldehydes and ketones

300 Column: Acclaim Carbonyl RSLC, 2.2μm 10 11 Dimension: 2.1 × 150mm 9 12 8 Mobile Phases: A: D.I. water B: Acetonitrile 1 7 Gradient (min): -4.5 0.0 8.3 15.0 18.0 %A 48 48 48 0 0 6 %B 52 52 52 100 100 2 Flow Rate: 0.400mL/min Injection: 1μL 5 Temperature: 28°C 3 Detection: UV at 360nm 4 (data collection rate at 25Hz) Samples: Calibration mixes diluted in methanol Peaks: 1. Formaldehyde DNPH mAU EPA 2. Acetaldehyde DNPH 3. Propionaldehyde DNPH 554 4. Crotonaldehyde DNPH 5. Butyraldehyde DNPH 13 14 15 16 17 19 6. Cyclohexanone DNPH 7. Valeraldehyde DNPH CARB 8. Hexanal DNPH 1004 9. Heptanal DNPH 10. Octanal DNPH 11. Nonanal DNPH 18 19 20 12. Decanal DNPH EPA 13. Acetone DNPH TO-11 14. Acrolein DNPH 15. Butanone DNPH 0 16. Methacrolein DNPH 0 5 10 15 18 17. Benzaldehyde DNPH 18. Isovaleraldehyde DNPH Minutes 19. Tolualdehyde DNPH 20. Xylylaldehyde DNPH www.thermoscienti c.com/chromatography 4-101 4 -102 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • specifically fortheanalysisofpolyaromatichydrocarbons(PAHs) Specially tailoredalkylbondedsilicawithahighcarbonloading,designed Green PAH Hypersil Available in3µmand5µmparticlesizevarietyofcolumndimensions Rapid analysisof16PAHs in4minutesusingshort,fastcolumns Optimized forEPA Method610 5 3 Particle Size (µm) Size Particle Guide Ordering PAH Green Hypersil HPLC Column Column HPLC Guard Cartridge Guard HPLC Column HPLC HPLC Column HPLC Guard Cartridge Guard Format 250 150 100 10 150 100 10 Length (mm) ID (mm) ID (mm) Length 4.6 4.6 4.6 4.6 4.6 2.1 4.6 3.0 2.1 4.6 3.0 2.1 www.youtube.com/chromatographyvideos chromatography channelonYouTube Subscribe toour 31105-254630 31105-154630 31105-104630 31105-014001 31103-154630 31103-152130 31103-104630 31103-103030 31103-102130 31103-014001 31103-013001 31103-012101 Cat. No. Cat. Dionex OmniPac LC Columns and Accessories DVB polymer columns for combined ion exchange and reversed-phase separations

• Acid, base and solvent compatible, pH 0 to 14 • Ideal for the separation of high molecular weight organic acids • Delivers optimal separation of very hydrophobic anions • Delivers optimal separation of halogenated anions • Provides simultaneous separation of neutral and ionic species • Unique selectivity for polar and ionic organic analytes • Delivers optimal separation of organic, hydrophobic, and halogenated cations

Thermo Scientific™ Dionex™ OmniPac™ PAX-100 columns are used to separate hydrophobic anionic analytes such as larger organic acids. The Dionex OmniPac PAX-500 column simultaneously separates anionic and neutral species. The Dionex OmniPac PCX-100 column separates low-molecular-weight hydrophobic cations. The Dionex OmniPac PCX-500 column simultaneously separates cationic and neutral >> species in a single run. Columns for Analytical LC The Dionex OmniPac PAX- and PCX-100 and 500 are latex-based columns. Both PAX columns have an ion exchange capacity of about 40 μeq per column, providing equivalent anion exchange separations. The PCX columns have a capacity of approximately 120 μeq per column. The PAX- and PCX-500 columns separate analytes through both ion exchange and reversed-phase mechanisms, due to their higher reversed-phase capacity relative to the PAX- and PCX-100 columns.

Dionex OmniPac Anion Exchange Ordering Guide Description Porosity Length (mm) 4.0mm ID PAX-100 microporous 50 042151 250 042150 PAX-500 macroporous 50 042153 250 042152

Dionex OmniPac Cation Exchange Ordering Guide Description Porosity Length (mm) 4.0mm ID PCX-100 microporous 50 042193 250 042189 PCX-500 macroporous 50 042195 250 042191

Gradient separation of nitrogen-containing compounds 16 Column: Dionex OmniPac PCX-500 (4 × 250mm) 12 Eluent: Acetonitrile/Sodium Chloride/Hydrochloric Acid Gradient 13 9 Flow Rate: 1.0mL/min Detection: UV, 254nm 6,7 Peaks: 1. Orotic Acid 2. 4-Hydroxybenzamide 1 2 4,5 3. Luminol Impurity 10 11 4. Luminol 5. Pyridine 17 6. PABA 15 7. 2,2’-Bipyridine 8 14 3 8. p-Phenylenediamine 9. Naphthylamine 10. Nitrobenzoic Acid 11. Tribenzylamine 0 5110 5 12. p-Nitroaniline Minutes 5410 13. 2,4-Dinitroaniline 14. Dibenzylamine 15. N-Methyl-N-nitrosoaniline 16. 4-Chloro-2-nitroaniline 17. 2,6-Dichloro-4-nitroaniline

www.thermoscienti c.com/chromatography 4-103 4 -104 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 01 • • • • • • The ThermoScientific of Hydrophobic,IonizableCompounds Polymeric Reversed-PhaseColumnIdealfortheSeparation NS2 and IonPac NS1 Dionex Gradient separationofinositolmono-,di-,tri-,andtetraphosphates performance analysisforhydrophobicaminesandacidsusingsuppressed conductivitydetection. trifluoroacetic acid(TFA), heptafluorobutyricacid(HFBA),ortetrabutylammoniumborate (TBAB).Itprovideshigh IonPac NS2isasilica-basedcolumnformobile-phaseionchromatography(MPIC)applications usingeluentscontaining column ofchoiceforroutineionpairchromatography. to solvents,acids,andbases,permitstheuseofeluentfrompH014.The DionexIonPacNS1columnisthe high-surface-area, ethylvinylbenzenepolymercrosslinkedwith55%divinylbenzene. ThisresinmakestheNS1resistant Utilizeion-pairchromatographyfordifficultseparations Canalsobeusedfortraditionalpolymericreversed-phaseapplications Compatiblewithacids,bases,andsolventfrompH0to14 Idealforseparationoflargemoleculesthatcarrylocalizedcharges,suchassurfactants Goodpeakshape Excellentresolution IonPac NS2 IonPac IonPac NS1IonPac Description Particle Size (µm) Size Particle Description Guide Ordering NS2 and NS1 IonPac Dionex 1 2 02 3 03 5 10 5 ™ Dionex Minutes 04 4 ™ 5 IonPac 6 05 ™ NS1-10μmandNS1-5μmcolumnsarepackedwithaneutral,macroporous, HPLC Column HPLC Guard Column HPLC Column HPLC Guard Column HPLC Column HPLC Format 7 8 9 06 10 5678 0 250 150 35 250 35 150 Length (mm) Length P (2) 1. Ins Suppressed Conductivity Peaks: 1.0mL/min Detection: Rate: Flow Gradient Hydroxide /Sodium Isopropanol Eluent: x250mm) (4 PAX-100 OmniPac Dionex Column: 10. Ins (1,4,5,6) P4 (1,4,5,6) Ins 10. P3 (2,4,5) Ins 9. P3 (1,4,5) Ins 8. P3 7. (1,3,4) Ins P2 (4,5) Ins 6. P2 (2,4) Ins 5. P2 (1,4) Ins 4. P (4) Ins 3. (1) P Ins 2. (Baseline Subtracted) – – – SP4354 SP4356 – 2.0mm ID 4.0mm ID 4.0mm ID 2.0mm SP6905 SP6906 SP6907 035321 039567 039568 Hypersil BDS LC Columns and Accessories A good choice for robust, general-purpose columns

Hypersil BDS Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) C18 C8 Phenyl CN 2.4 Drop-in Guard 10 2.1 28102-012101 28202-012101 – – (4/pk) 4.0/4.6 28102-014001 28202-014001 – – 50 2.1 28102-052130 28202-052130 – – 4.6 28102-054630 28202-054630 – – 100 2.1 28102-102130 28202-102130 – – 4.6 28102-104630 28202-104630 – – 150 2.1 28102-152130 28202-152130 – – 4.6 28102-154630 28202-154630 – – 3 Drop-in Guard 10 2.1 28103-012101 28203-012101 28903-012101 28803-012101 (4/pk) 3.0 28103-013001 28203-013001 – – 4.0/4.6 28103-014001 28203-014001 28903-014001 28803-014001 HPLC Column 30 2.1 28103-032130 – – – 50 2.1 28103-052130 28203-052130 28903-052130 28803-052130 >>

3.0 28103-053030 – – – Columns for Analytical LC 4.6 28103-054630 28203-054630 – – 100 2.1 28103-102130 – – – 3.0 28103-103030 – – – 4.0 28103-104030 – – – 4.6 28103-104630 28203-104630 – – 150 2.1 28103-152130 – – – 3.0 28103-153030 28203-153030 – – 4.0 28103-154030 – – – 4.6 28103-154630 28203-154630 28903-154630 28803-154630 5 Drop-in Guard 10 2.1 28105-012101 28205-012101 – – (4/pk) 3.0 28105-013001 28205-013001 – – 4.0/4.6 28105-014001 28205-014001 28905-014001 28805-014001 HPLC Column 50 2.1 28105-052130 28205-052130 – – 3.0 28105-053030 28205-053030 – – 4.6 28105-054630 28205-054630 – – 100 2.1 28105-102130 28205-102130 – – 3.0 28105-103030 – – – 4.0 28105-104030 – – – 4.6 28105-104630 28205-104630 – – 125 3.0 28105-123030 – – – 4.0 28105-124030 – – – 4.6 28105-124630 – – – 150 2.1 28105-152130 – – – 3.0 28105-153030 – – – 4.0 28105-154030 28205-154030 – – 4.6 28105-154630 28205-154630 28905-154630 28805-154630 250 2.1 28105-252130 – – – 3.0 28105-253030 – – – 4.0 28105-254030 28205-254030 28905-254030 – 4.6 28105-254630 28205-254630 28905-254630 28805-254630 Format Length (mm) ID (mm) Cat. No. UNIGUARD Guard 10 1.0 851-00 Cartridge Holder 2.1 852-00 3.0 852-00 4.0/4.6 850-00

www.thermoscienti c.com/chromatography 4-105 4 -106 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories A globalstandardformanyexistingmethods ClassicalHypersil Particle Size (µm) Size Particle Guide Ordering ODS Hypersil 3 5 Format Drop-in Guard (4/pk) HPLC Column HPLC Drop-in Guard (4/pk) HPLC Column HPLC Length (mm) ID (mm) ID (mm) Length 10 50 100 125 150 250 10 50 100 125 150 200 250 4.0/4.6 3.0 2.1 4.6 3.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 3.0 2.1 4.0/4.6 3.0 2.1 4.6 3.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 3.0 2.1 4.6 4.0 2.1 4.6 4.0 3.0 2.1 Cat. No. Cat. 30103-014001 30103-013001 30103-012101 30103-054630 30103-053030 30103-052130 30103-104630 30103-104030 30103-103030 30103-102130 30103-124630 30103-124030 30103-122130 30103-154630 30103-154030 30103-153030 30103-152130 30103-254630 30103-254030 30103-253030 30103-252130 30105-014001 30105-013001 30105-012101 30105-054630 30105-053030 30105-052130 30105-104630 30105-104030 30105-103030 30105-102130 30105-124630 30105-124030 30105-123030 30105-122130 30105-154630 30105-154030 30105-153030 30105-152130 30105-204630 30105-204030 30105-202130 30105-254630 30105-254030 30105-253030 30105-252130 LC Columns and Accessories

Hypersil ODS-2 Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 3 Drop-in Guard (4/pk) 10 4.0/4.6 31603-014001 HPLC Column 50 4.6 31603-054630 100 4.0 31603-104030 4.6 31603-104630 150 4.6 31603-154630 5 Drop-in Guard (4/pk) 10 4.0/4.6 31605-014001 HPLC Column 50 4.6 31605-054630 100 4.6 31605-104630 150 4.6 31605-154630 250 4.0 31605-254030 4.6 31605-254630

>> Hypersil MOS (C8) Ordering Guide Columns for Analytical LC Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 5 Drop-in Guard (4/pk) 10 4.0/4.6 30205-014001 HPLC Column 50 4.6 30205-054630 100 4.6 30205-104630 150 4.6 30205-154630 250 4.0 30205-254030 4.6 30205-254630

Hypersil MOS-2 (C8) Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 5 Drop-in Guard (4/pk) 10 4.0/4.6 30305-014001 HPLC Column 150 4.6 30305-154630 250 4.0 30305-254030 4.6 30305-254630

Hypersil SAS (C1) Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 5 Drop-in Guard (4/pk) 10 4.0/4.6 30505-014001 HPLC Column 150 4.6 30505-154630 250 4.6 30505-254630

www.thermoscienti c.com/chromatography 4-107 4 -108 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Hypersil ClassicalHypersil Particle Size (µm) Size Particle Guide Ordering APS-2 Hypersil (µm) Size Particle Guide Ordering CPS-2 Hypersil (µm) Size Particle Guide Ordering CPS Hypersil (µm) Size Particle Guide Ordering Phenyl-2 Hypersil (µm) Size Particle Guide Ordering Phenyl Hypersil 3 5 3 5 5 5 5 Format Format Format Format Format Drop-in Guard (4/pk) Drop-in Guard (4/pk) Drop-in Guard (4/pk) Drop-in Guard (4/pk) Drop-in Guard (4/pk) HPLC Column HPLC Column HPLC Column HPLC Column HPLC Column HPLC Drop-in Guard (4/pk) HPLC Column HPLC Drop-in Guard (4/pk) HPLC Column HPLC continued Length (mm) ID (mm) ID (mm) Length (mm) ID (mm) Length (mm) ID (mm) Length (mm) ID (mm) Length (mm) ID (mm) Length 10 10 10 10 10 50 150 150 150 50 10 100 250 250 100 150 150 150 250 10 250 100 150 250 4.0/4.6 2.1 4.0/4.6 4.0/4.6 4.0/4.6 4.0/4.6 4.6 2.1 4.6 4.6 4.6 4.6 4.0/4.6 2.1 4.6 4.6 4.6 4.6 4.6 2.1 4.6 4.6 4.0 4.0/4.6 3.0 4.6 4.0 4.6 3.0 4.6 4.6 4.0 Cat. No. Cat. No. Cat. No. Cat. No. Cat. No. Cat. 30703-014001 30703-012101 31905-014001 30803-014001 31905-014001 30905-014001 30703-054630 30703-052130 31905-154630 30803-154630 31905-154630 30905-054630 30805-014001 30703-102130 31905-254630 31905-254630 30905-104630 30805-154630 30703-154630 30703-152130 30905-154630 30805-254630 30805-254030 30705-014001 30705-013001 30905-254630 30905-254030 30705-104630 30705-103030 30705-154630 30705-254630 30705-254030 LC Columns and Accessories

Hypersil Silica Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 3 Drop-in Guard (4/pk) 10 2.1 30003-012101 3.0 30003-013001 4.0/4.6 30003-014001 HPLC Column 50 2.1 30003-052130 3.0 30003-053030 4.6 30003-054630 150 4.6 30003-154630 5 Drop-in Guard (4/pk) 10 3.0 30005-013001 4.0/4.6 30005-014001 HPLC Column 50 4.6 30005-054630 100 4.6 30005-104630 150 4.6 30005-154630

250 4.0 30005-254030

4.6 30005-254630 >> Columns for Analytical LC Hypersil SAX Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. 5 Drop-in Guard (4/pk) 10 2.1 34105-012101 3.0 34105-013001 4.0/4.6 34105-014001 HPLC Column 50 2.1 34105-052130 150 4.6 34105-154630 250 3.0 34105-253030 4.6 34105-254630

Other HPLC Columns

Aquasil HPLC Columns Thermo Scientific™ Aquasil™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details.

BetaBasic HPLC Columns Thermo Scientific™ BetaBasic™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details.

BETASIL HPLC Columns Thermo Scientific™ BETASIL™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details.

HyPurity HPLC Columns Thermo Scientific™ HyPurity™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details.

www.thermoscienti c.com/chromatography 4-109 4 -110 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Thermo Scientific for fasterseparationanalysis. shallower gradientsonshortercolumns, particles supportincreasedflowratesand Compared to5μmcolumnpackings,thesmaller for rapidanalysisofcomplexproteindigests. Acclaim 300 C18 features 3μmsilicaparticles and C4, chemistries. reversed phase packings areavailableinC18,C8 reliable results.BioBasic for surface reproducible stable, highly a produce used forthesepackings dense bonding chemistry extra the because columns providesuperiorchromatography Thermo Scientific Ion Exchange for fastproteinseparationanalysis. highhigh resolutionatexceptionally flow rates columns uniquelyprovidethe advantages of Thermo Scientific Reversed Phase Columns forproteins of proteins,peptides,oligonucleotidesandotherbiomolecules. and polymericcolumnsspecificallydesignedtohandletheuniquerigors oftheanalysis characterization. ThermoScientificaddressestheseneedswitha rangeofsilica chemistries, columnconfigurationsanddetectiontechniquesfor their effective coupled withmatrixcomplexitydemandsarangeofsampleseparation modes,column The diversityofbiologicalsamplesintermscompoundstructure andproperties Thermo Scienti c LCColumnsforBiomolecules monoclonal antibodyvariants. specifically designedfortheanalysis of diary products.MAbPaccolumnsare matrices, suchasbiopharmaceuticalsand include proteinvariantsinavarietyof analytical characterization.Applications developed formonoclonalseparationand MAbPac SCXcolumnsarespecifically and and moderatecapacity.WCX ProPac separation and fastmasstransferforhigh-efficiency controlsurface providespH-basedselectivity interactions, andagraftedcationexchange A hydrophiliclayerpreventsunwantedsecondary residue. isoforms thatdifferbyasinglecharged for separationsofproteinvariants,resolving providing exceptionallyhighresolutionandefficiency based onapellicularnonporouscoreparticles Scientific ™ MAbPac ™ ™ ™ ProSwift BioBasic ProPac ™ ion exchange columns are ion exchangecolumnsare ™ andThermo ™ ™ reversed-phase RPmonolith the highlyaqueousconditionsused in HIC. provide exceptionalhydrolyticstabilityunder preparative applications.ProPacHICcolumns of proteinsandvariantsforanalytical that providesexcellenthighresolutionseparations high-capacity, 5µmsilica-basedHICcolumn 300Å, The ProPacHIC-10columnisahigh-resolution, Hydrophobic Interaction Fab andFcfragmentsresultingfromproteolysis. and theiraggregates,aswelltheanalysisof characterization ofmonoclonalantibodies(MAb) specifically designedforseparationand exclusion chromatography(SEC)column MAbPac SEC-1(300Å 5µmsilica)isasize used forsampleclean-uppriortootheranalyses. and watersolublepolymers.Theycanalsobe proteins molecular weightdeterminationofpeptides, 1000Å) areavailable,makingthemidealfor 300Å, 120Å, based columns.Fourporesizes(60Å, with higherefficienciesthanthatofpolymer- stability ofsilica-basedsizeexclusioncolumns proprietary polymericcoating,offerthemechanical BioBasic SECcolumns,basedonsilicawitha Size Exclusion to traditionalporousionexchangemedia. combined withexcellentresolutioncompared loading capacitycomparedtopellicularphases ion exchangemedia.Theyofferincreased excellent alternativetoporousornon ProSwift ionexchangemonolithsprovidean conditions. exchange ion acids usingproteinfriendly peptides and nucleic proteins, for performance efficiency. Bothphasesprovidesuperior because the5μm,300Åsilicaprovideshigh both column-to-columnandbatch-to-batch columns demonstratesuperiorreproducibility, BioBasic AXandSCXionexchange containing high-mannoseregions. glycans, glycopeptides,andglycoproteins and purificationofConcanavalinA(ConA)binding is unsurpassedforfast,highlyefficientenrichment The ProSwiftConA-1Saffinitymonolithcolumn layer, thengraftedwithpoly(IDA)chains. polymeric beadscoatedwithahydrophilic chromatography.nonporous, 10μm, Itispackedwith affinity metal proteins andpeptidesbyimmobilized semipreparative columnforseparation of and is ahigh-resolutionanalytical IMAC-10 titer analysisfromharvestcellculture.TheProPac accurate determinationofmonoclonalantibody porous polymericcolumndesignedforfast, The MAbPacProteinAcolumnisauniquenon- Affinity Thermo Scientific possible withothercolumns. n–1, n+1,andotherfailurenot sequences can resolvefulllengtholigonucleotides from synthetic oligonucleotides.DNAPaccolumns resolution foranalysisandpurificationof industry-leading exchange columnsprovide flourescently labeledornativeglycans. by charge,sizeandpolarity. Separatingeither for thesimultaneousseparationofglycans AXR-1 columnsareHPLCdesigned Thermo Scientific Columns forcarbohydrates Thermo Scientific Columns foroligonucleotides column, withincreasedloadingcapacity. resolution andselectivityoftheDNAPac semipreparative columnincorporatesthehigh exceptionally higholigonucleotidepurity. This exchange monolithcolumnthatprovides ™ ™ ™ DNAPac GlycanPac DNASwift ™ strong anion stronganion ™ ™ astronganion AXH-1and LC Columns and Accessories Columns for proteomics before LC separation with MS detection. they deliver outstanding chromatographic Nano, capillary and micro columns The columns are designed to provide the highest performance on any nano LC system. Thermo Scientific™ Acclaim™ PepMap™ and efficiencies for one dimensional peptide Thermo Scientific™ EASY-Spray™ columns PepMap RSLC columns are specially designed for mapping experiments and 2D-LC analyses. provide a fully integrated and temperature ™ ™ high-resolution analyses of tryptic, natural, and Thermo Scientific PepSwift monolithic controlled combined column-emitter design synthetic peptides. The columns are often columns are specially designed for fast with only a single nanoViper connection applied for LC-MS/MS peptide mapping for separation and identification of proteins between the LC and the MS ion source. This protein identification, biomarker discovery, and and peptides using nano and capillary LC dead volume reduction is a critical component systems biology. Due to their high loading coupled to MS. in helping to deliver state of the art capacity, the columns are exceptionally suitable Using highly pure chromatographic media and performance with ease of use. for the analysis of low abundant peptides in biocompatible, metal-free fused silica capillaries, complex proteomics samples. Acclaim PepMap Thermo Scientific™ EASY-Column™ capillary LC Trap columns are typically applied for the columns are produced with a focus on design desalting and preconcentration of peptides simplicity and strict quality control. As a result,

Bio Columns Selection Guide >> Columns for Biomolecules

Analyte Mode of Analysis Recommended Column

BioBasic SEC 4-139 Size Exclusion MAbPac SEC-1 4-129 BioBasic AX 4-141 ProPac SCX-10, WCX-10, SAX-10, WAX-10, SCX-20 4-116 Ion Exchange MAbPac SCX-10, MAbPac SCX-10 RS 4-127 Monoclonal ProSwift IEX 4-123 antibodies and BioBasic 18, 8, 4 4-140 Proteins Acclaim 300 C18 4-142 Reversed Phase View product information Accucore 150-C18, 150-C4 4-144 ProSwift RP 4-124 and application notes Hydrophobic Interaction ProPac HIC-10 4-121 MAbPac Protein A 4-126 Af nity ProPac IMAC-10 4-122 ProSwift ConA-1S 4-125 Acclaim PepMap 4-149 PepSwift 4-154 Proteomics EASY-Column 4-155 Peptides EASY-Spray Columns 4-153 BioBasic 18, 8, 4 4-140 Analytical Acclaim 300 4-142 Preparative BioBasic Inquire Amino Acids Ion Exchange AminoPac PA10 Inquire (derivatized) Reversed phase Hypersil GOLD 4-026 Amino Acids Ion Exchange AminoPac PA10 Inquire (underivatized) Reversed phase Hypercarb 4-084 Anion Exchange BioBasic AX 4-141 Nucleotides Polar retention Hypercarb 4-084 BioBasic AX 4-141 Oligonucleotides Ion Exchange DNAPac PA100, PA200, PA200 RS 4-134 DNASwift 4-137 Ligand Exchange HyperREZ XP 4-056 Ion Exchange CarboPac Inquire Mixed Mode GlycanPac AXH-1 4-032 Acclaim HILIC 4-067 Carbohydrates Hypersil GOLD HILIC 4-037 HILIC Syncronis HILIC 4-081 Accucore 150-Amide-HILIC 4-146 Polar retention Hypercarb 4-084

www.thermoscienti c.com/chromatography 4-111 4 -112 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Silica-based Hydrophobic Interaction Chromatography Phases Chromatography Silica-based Hydrophobic Interaction Phases Chromatography Exclusion Size Silica-based Separations Protein for Columns Phases Exchange Ion and Phase Reversed Silica-based Phases Biomolecules for HPLC SCX AX 4 8 18 Phases BioBasic 150-Amide-HILIC 150-C4 150-C18 Accucore Phases 300 C4 300 100 C8 300 C18 300 100 C18 100 Acclaim PepMap Phases Column SEC 1000 300 SEC 120 SEC 60 SEC Phases BioBasic Phase Phase 300 C18 300 Acclaim Phase HIC-10 ProPac Phase Interaction Interaction Hydrophobic Aqueous Aqueous Aqueous Aqueous SEC Type peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, solid core silica core solid Spherical, Spherical, solid core silica core solid Spherical, Spherical, solid core silica core solid Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, fully porous silica 3, 5 3, silica porous fully Spherical, peia,flypru iia5 silica porous fully Spherical, Spherical, fully porous silica 2, 3, 5 3, 2, silica porous fully Spherical, Particle Type Particle peia,flypru iia3 silica porous fully Spherical, Target Applications protein variants protein proteins and of separations resolution High peia,flypru iia5 silica porous fully Spherical, 5 silica porous fully Spherical, 5 silica porous fully Spherical, 5 silica porous fully Spherical, Type Particle Material Base Matrix surface chemistry surface amide/ethyl with silica ultrapure Spherical, porous 2.6 2.6 2.6 (µm) Size Particle (µm) Size Particle Size (µm) Size Particle 5 www.thermoscienti c.com/chromatography thousands ofapplications Search ourdatabaseof 300 300 300 300 300 150 150 150 0 300 300 100 300 100 Size (Å) Size Pore 300 Size (Å) Size Pore 300 1000 300 120 60 (Å) Size Pore 100 100 100 100 100 80 80 80 300 100 300 Area (m Nominal Surface 100 Area (m Nominal Surface 100 20 –4000 20 1 –500 0.1 –50 0.1 –6 Operating Range (kDa) Limit Exclusion 2 /g) 2 /g) 3 3 4 5 9 – – – 2 7 3 9 9 15 8 Carbon % column 7.8per x75mm lysozyme 340mg Capacity Breakthrough No No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Endcapping L33 L59 L33, L33 – Code USP L52 – L26 L7 L1 L26 L1 L26 L7 L1 L1 L1 Code USP solvent for cleanup for solvent organic salts, sulfate/phosphate Ammonium Compatibility Solvent 733 731 723 736 735 734 733 Code Phase 722 721 167 165 161 – – – – – Code Phase 4 4 4 4 4 4 4 Page 4 4 4 4 4 4 4 4 4 4 Page -141 -141 -140 -139 -139 -139 -139 -140 -140 -146 -145 -144 -149 -149 -149 -149 -142 2.5-7.5 Range pH Polymeric Ion Exchange, Reversed Phase and Af nity Columns LC Columns and Accessories

Target Base Matrix Functional Breakthrough Recommended Solvent Maximum pH Column Phase Applications Material Groups Capacity Flow Rate Compatibility Backpressure Range ProPac Weak High resolution Ethylvinylbenzene Carboxylate 6mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 WCX-10 Cation separations of cross linked with lysozyme mL/min acetone. (21 MPa) Exchange proteins and protein 55% divinylbenzene Incompatable variants 10µm nonporous with alcohols particles and MeOH ProPac Strong High resolution Ethylvinylbenzene Sulfonate 3mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 SCX-10 Cation separations of cross linked with lysozyme mL/min acetone, (21 MPa) Exchange proteins and protein 55% divinylbenzene MeOH variants 10µm nonporous particles ProPac Strong High Resolution Divinlybenzene Sulfonic 20μg/mL 0.2-2.0 50% 3000psi 2.0-12 SCX-20 Cation separations of 10μm nonporous Dynamic mL/min acetonitrile (21 MPa) Exchange proteins and protein particles capacity variants ProPac Weak High resolution Ethylvinylbenzene Tertiary 5mg/mL BSA 0.2-2.0 80% ACN, 3000psi 2.0-12 WAX-10 Anion separations of cross linked with amine mL/min acetone, (21 MPa) Exchange proteins and protein 55% divinylbenzene MeOH,

variants 10μm nonporous particles >> ProPac Strong High resolution Ethylvinylbenzene Quaternary 15mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 SAX-10 Anion separations of cross linked with ammonium BSA mL/min acetone, (21 MPa) Columns for Biomolecules Exchange proteins and protein 55% divinylbenzene MeOH variants 10µm nonporous particles ProSwift Reversed Fast protein analysis Monolith; Phenyl 5.5mg/mL 2.0-4 .0 Most common 2800psi 1.0-14 RP-1S Phase with high resolution polystyrene- Insulin mL/min organic (19.2 MPa) of large peptides to divinylbenzene solvents medium proteins ProSwift Reversed Fast protein analysis Monolith; Phenyl 1.0mg/mL 1.0-10 Most common 2800psi 1.0-14 RP-2H Phase with high resolution polystyrene- Lysozyme mL/min organic (19.3 MPa) over a wide size range divinylbenzene solvents ProSwift Reversed Fast protein analysis Monolith; Phenyl 0.5mg/mL 1.0-16 Most common 2800psi 1.0-14 RP-3U Phase with high resolution polystyrene- Lysozyme mL/min organic (19.3 MPa) of large proteins divinylbenzene solvents ProSwift Reversed Fast protein analysis Monolith; Phenyl 2.3mg/mL 0.1-0.3 Most common 1500psi 1.0-14 RP-4H Phase with high resolution polystyrene- Lysozyme mL/min organic divinylbenzene Solvents ProSwift Strong Fast protein analysis Monolith; Quaternary 18mg/mL 0.5-1.5 Most common 1000psi 2.0-12 SAX-1S Anion with high resolution polymethacrylate amine BSA (4.6mm) organic (4.6mm) Exchange solvents 2000psi (1.0mm) ProSwift Strong Fast protein analysis Monolith; Sulfonic 30mg/mL 0.5-1.5 Most common 1000psi 2.0-12 SCX-1S Cation with high resolution polymethacrylate acid Lysozyme mL/min organic (4.6mm) Exchange (4.6mm) solvents ProSwift Weak Fast protein analysis Monolith; Tertiary 18mg/mL 0.5-1.5 Most common 1000psi 2.0-12 WAX-1S Anion with high resolution polymethacrylate amine BSA mL/min organic (4.6mm) Exchange (DEAE) (4.6mm) solvents 2000psi (1.0mm) ProSwift Weak Fast protein analysis Monolith; Carboxylic 23mg/mL 0.5-1.5mL/min Most common 1000psi 2.0-12 WCX-1S Cation with high resolution polymethacrylate acid Lysozyme (4.6mm), organic (4.6mm) Exchange 0.05-0.20 solvents 2000psi (1.0mm) ProPac Immobilized High resolution Polystyrene Poly (IDA) >60mg 1.0mL/min EtOH, urea, 3000psi 2.0-12 IMAC-10 Metal separation of divinylbenzene grafts lysozyme/mL NaCl, non-ionic (21MPa) Af nity certain metal- 10μm nonporous gel detergents, binding proteins particles (4 x 250mm) glycerol, and peptides acetic acid, guanidine HCl ProSwift Af nity Concanavalin A Monolith; Concanavalin 12-16mg of 0-1.0mL/min Up to 10% 2000psi 5.0-8 ConA-1S binding (high- polymethacrylate A ligands protein methanol mannose) glycans, glycopeptides and proteins

www.thermoscienti c.com/chromatography 4-113 4 -114 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Silica-based HILIC and Mixed-Mode Columns and Mixed-Mode Silica-based HILIC Columns Exchange Ligand Polymeric Separations Carbohydrate for Columns Separations Monoclonal for Antibody Columns Phase Phase Af Columnsnity Protein A Protein MAbPac Sugar Alcohol 150-Amide-HILIC Accucore Phase Organic Acid Organic Carbohydrate Na Carbohydrate Ca Carbohydrate Pb AXH-1 Phase Phase Phase Columns Chromatography Exclusion Size Silica-based Phase Columns Exchange Ion Polymeric AXH-1 GlycanPac Phases Carbohydrate H Phases XP HyperREZ MAbPac SEC-1 WCX-10 ProPac SCX-10 MAbPac Af ligandnity Protein A Protein IEX Type IEX (carboxylate) exchange cation Weak (sulfonic) exchange cationStrong + 2+ 2+ + SEC Type Aqueous Spherical, polymer Spherical, solid core silica core solid Spherical, Spherical, polymer peia,flypru iia3 silica porous fully Spherical, Particle Type Particle Spherical, polymer Spherical, polymer Spherical, polymer Spherical, polymer Type Particle Spherical, fully porous silica porous fully Spherical, Particle Type Type Particle Polymeric Particle Type Type Particle Non-porous DVB Polymeric, crosslinked DVB Polymeric, Highly Particle Type Particle Size (μm) Pore Size (Å) Size Pore (μm) Size Particle Type Particle porous silica porous fully Spherical, Size (μm) Size Particle 10 10 5, 3, Size (μm) Size Particle 12 2.6 10 8 8 (µm) Size Particle 8 8 8 (µm) Size Particle 1.9 5 (Å) Size Pore non-porous non-porous (Å) Size Pore non-porous – – 150 – – 120 Size (Å) Size Pore – – – – – – (Å) Size Pore – – 175 Capacity Capacity Dynamic 6mg/mL µg/mL 40 5µm: SCX-10 RS MAbPac 20µg/mL 10µm: 40µg/mL 5µm: 60µg/mL 3µm: PEEK SCX-10 MAbPac Capacity Capacity column column IgG/ 100µg 300 80 300 Area (m Nominal Surface Area (m Nominal Surface 220 Flow Rate Rate Flow Recommended < 2.5 Flow Rate Rate Flow Recommended pressure limits) pressure column and size particle the (Depending on 0.2-2.0mL/min 0.2-2.0mL/min 2 2 /g) /g) Range (kDa) Range Operating Limit Exclusion 1-500 – – - - – – - – – – – – – – – – - Carbon % Carbon % temperature Maximum 30 Compatibility Compatibility Solvent Acetonitrile 50% methanol including alcohols with Incompatible acetone. ACN, 80% Yes Yes Endcapping Endcapping Backpressure Maximum 1000 Backpressure Maximum 7000psi columns RS (21MPa) 3000psi (21MPa) 3000psi L19 - L17 - - - L19 L34 L17 - - Code USP Code USP USP Code USP L33, L59 L33, 697 167 696 692 691 690 693 Code Phase Code Phase Range Range pH 2.5-7.5 Range Range pH 2.0-12 2.0-12 2.0-12 4 4 4 4 4 4 4 4 4 Page Page -156 -156 -156 -132 -156 -156 -156 -156 -132 Page 4 Page 4 Page 4 -126 -127 -129 Monolithic Af nity Columns LC Columns and Accessories Particle Size Pore Nominal Surface % USP Phase Phase Particle Type Endcapping Page (µm) Size (Å) Area (m2/g) Carbon Code Code ProSwift Phases ConA-1S Concanavalin A binding (high "Monolith; 175 220 - Yes - - 4-125 mannose) glycans, glycopeptides and proteins

Columns for Oligonucleotide Separations Column Target Applications Matrix Base Material Substrate Crosslinking Latex Crosslinking Capacity Recommended Eluents Recommended Rate Flow Solvent Compatibility Maximum Backpressure pH Range DNAPac High resolution 13µm diameter 55% 5% 40µeq Hydroxide or 1.5mL/min 0-100% 4000psi 2-12.5 PA100 separations of nonporous substrate sodium and (28MPa)

single and double agglomerated with lithium salts >> stranded DNA or alkyl quaternary of Chloride or RNA ammonium Perchlorate Columns for Biomolecules oligonucleotides functionalized latex 100nm MicroBeads DNAPac Improved high 8µm diameter 55% 5% 40µeq Hydroxide, 1.2mL/min 0-100% 4000psi 2-12.5 PA200 resolution nonporous substrate acetate/ or (28MPa) separations of agglomerated with sodium and single and double alkyl quaternary lithium salts stranded DNA or ammonium of Chloride or RNA orthogonal to functionalized latex Perchlorate DNAPac PA100 130nm MicroBeads DNAPac UHPLC-resolution 4µm diameter 55% 5% 40µeq Hydroxide, 1.3mL/min 0-100% 10,000 2-12.5 PA200 separations of nonporous substrate acetate/ psi RS single and double agglomerated with hydroxide or (69MPa) stranded DNA or alkyl quaternary sodium and RNA Best ammonium lithium salts available functionalized latex of Chloride or resolution 130nm MicroBeads Perchlorate

DNASwift High resolution Monolith; 70% 3% 8mg, NaClO4 and 0.5-2.5mL Most 1500psi, 6.0-12.4 separations for polymethacrylate of a 20 mer NaCl Common 10MPa puri cation of substrate agglomerated oligonucleotide Organic oligonucleotides with quaternary amine Solvents Highest functionalized latex latex-based capacity

www.thermoscienti c.com/chromatography 4-115 4 -116 -116 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories control andfastmasstransferforhigh-efficiencyseparationmoderatecapacity. selectivity prevents unwantedsecondaryinteractions,andagraftedcationexchangesurfaceprovidespH-based layer ProPac WCX-10andSCX-10columnscanresolveisoformsthatdifferbyasinglechargedresidue.Ahydrophilic • • • • • • • providing exceptionally highresolutionandefficiencyforseparations ofproteinvariants Weak andstrongcationexchangecolumnsarebasedonanonporouscoreparticle ProPac WCX-10 and SCX-10 ProPac SCX-10isastrongcationexchangecolumnwithsulfonatefunctionalgroups ProPac WCX-10isaweakcationexchangerwithcarboxylatefunctionalgroups Useful forcharacterizationofrelatedproteinvariantsincludingdeamidationandMAblysine Unmatched column-to-columnandlot-to-lotreproducibility High-ef ciencypeaks Unequalled highresolutionseparations Characterization andqualitycontrolassessmentofmonoclonalantibodiesotherproteins 10 10 Particle Size (µm) Size Particle Guide Ordering SCX-10 ProPac (µm) Size Particle Guide Ordering WCX-10 ProPac truncation variants HPLC Column HPLC Column HPLC Guard Column Guard Column Format Format 250 150 100 250 50 50 50 Length (mm) Length (mm) Length 063472 – – 063456 – 063462 063480 2.0mm ID 2.0mm ID 2.0mm 054993 SP6703 SP5829 054995 074600 079930 054994 4.0mm ID 4.0mm ID 4.0mm 063474 – – 063700 – – – 9.0mm ID9.0mm ID9.0mm SP5482 – – SP5522 – – – 22.0mm ID 22.0mm ID 22.0mm LC Columns and Accessories ProPac WCX-10 and SCX-10 continued

MAb lysine truncation variants

Column: ProPac WCX-10 , 4 x 250mm Lysine truncation Eluents: (E1) 20mM MES variants* + 115mM NaCl + 1mM EDTA, pH 5.5 K (E2) 20mM MES + 145mM NaCl KK + 1mM EDTA, pH 5.5 Gradient: t (min) %E1 %E2 0 100 0 AU MAb sample as received 2 100 0 40 0 100 60 0 100 Acidic charge Basic charge Sample: MAb variants variants Flow Rate: 1.0mL/min λ: 280nm * Peak assignment supported by data from R.J. Harris, After digestion with carboxypeptidase B et.al, J.Chromatogr., A 1995,705, 129–134. and Carboxypeptidase B digest. 0 10 20 30 40 Minutes 21104

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Hemoglobin variants Columns for Biomolecules

Column: ProPac SCX-10, 4 × 250mm 1 1.60 Eluent: A) 20mM Sodium phosphate, 4mM Potassium cyanide, pH 6 B) 1 M Sodium chloride in water C) Water 2 Gradient: Time %A %B %C 3 Init 50 0 50 AU 4 30 min 50 50 0 Flow Rate: 1mL/min Inj. Vol.: 10μL Detection: 220nm 0 Sample: 1. Fetal hemoglobin 0 5 10 15 20 25 2. Hemoglobin Minutes 14067 3. Sickle cell hemoglobin 4. Hemoglobin C

ProPac Kits Ordering Guide Part Number Phase Description Set Contents Column Dimensions SP5731 ProPac WAX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm SP5732 ProPac WAX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm SP5729 ProPac SAX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm SP5730 ProPac SAX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm SP5512 ProPac WCX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm SP5513 ProPac WCX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm SP5727 ProPac SCX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm SP5728 ProPac SCX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm

www.thermoscienti c.com/chromatography 4-117 4 -118 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Snake venomsfromnajaandRussell’s viper • • interactions betweenthebeadsurfaceandbiopolymer. compatible resinbeadsthatareuniformlycoatedwithahighlyhydrophiliclayer toreducenon-specific resolution separationsofproteins.Thestationaryphaseiscomposed10μm, non-porous, solvent The ProPacSCX-20columnisastrongcationexchange(SCX)designedspecifically toprovidehigh- Columns ProPac SCX-20 100 200 300 400 100 350 200 Fast masstransferforhigh-ef ciencyseparation Grafted cation-exchangesurfaceprovidespH-basedselectivitycontrol -50 -50 mAU mAU 10 Description (µm) Size Particle Guide Ordering SCX-20 ProPac 0 5 HPLC Column HPLC Guard Column Minutes 10 250 50 Length (mm) Length B A 1715 Samples: A. Snake Venom (Naja naja) 1mg/mL naja) (Naja Venom Snake A. 214nm UV, Samples: 30°C 10μL 10 min Bin 1–100% Detection: A Eluent in NaCl 0.5M Temperature: 7.3 pH Tris 20mM Volume: Inj. Gradient: B: Eluent A: Eluent 4×250mm SCX-20, ProPac Column: 074628 074643 4.0mm ID 4.0mm B. Snake Venom (Russell’s viper) viper) B. (Russell’s Venom Snake 1mg/mL LC Columns and Accessories ProPac WAX-10 and SAX-10 Weak and strong anion exchange columns are based on a nonporous core particle providing unequalled high resolution and efficiency in the separations of protein variants

• Unequalled resolution • High-ef ciency peaks • Useful for characterization and quality control assessment of closely-related protein variants • Supports separation of proteins that differ by as little as one amino acid residue • Neutral hydrophilic coat that eliminates protein-resin hydrophobic interactions • Superior lot-to-lot and column-to-column reproducibility • ProPac WAX-10 column is a weak anion exchanger with a tertiary amine functional group • ProPac SAX-10 is a strong anion exchange column with quaternary amine functional group

These columns can resolve protein isoforms that differ by as little as one charged residue. A hydrophilic layer prevents >> unwanted secondary interactions and a grafted anion exchange surface provides pH based selectivity control and fast mass transfer for high peak efficiency and resolution separations. Columns for Biomolecules

Resolution of phosphorylation variants of ovalbumin

Column: ProPac SAX-10, 4 x 250mm Eluents: A) Water B) 2.0 M NaCl C) 0.1 M Tris/HCl (pH 8.5) Gradient: Time %A %B %C Init 80 0 20

mAU 15 min 67.5 12.5 20 Flow Rate: 1.0mL/min Inj.Volume: 30μL Ovalbumin + alk. phos. Detection: 214nm Sample: Ovalbumin before and after Ovalbumin alkaline phosphatase treatment

0 2 4 6 8 10 12 14 16 Minutes 16445

Effect of sialylation on transferrin chromatography

Columns: ProPac SAX-10, 4 × 250mm Eluents: A) Water B) 2.0 M NaCl C) 0.2 M Tris/HCl (pH 9) HOLO Gradient: Time %A %B %C Init 87 3 10 30 min 83 7 10 mAU HOLO + neur Flow Rate: 1.0mL/min APO Lot #1 Inj. Volume: 50μL APO Lot #1+neur Detection: 214nm Samples: HOLO (iron rich) and APO (iron poor) human transferrin samples before and after neuraminidase treatment. Digestions were carried out APO Lot #2 overnight at 37 C in sodium acetate APO Lot #2 + neur buffer at pH 5. 0 4 8 12 16 20 24 28 32 Minutes 16446

www.thermoscienti c.com/chromatography 4-119 4 -120 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories

mAU 0 Gradient separationofproteinstandards The ProPacPA1 columnsupportstheanalysisandpurificationofhydrophilicanionicproteinspeptides. • • • • For hydrophilicanionicproteinseparations PA1ProPac 1 Pellicular packingensureshigh-ef ciencyandfastmasstransport Available insemipreparativeformat Ideal forhigh-resolutionseparationsofproteinswithpIvaluesfrom3to11 Good forhydrophilicproteinsandpeptides 10 10 Particle Size (µm) Size Particle Guide Ordering SAX-10 ProPac (µm) Size Particle Guide Ordering WAX-10 ProPac 10 Particle Size (µm) Size Particle Guide PA1 Ordering ProPac 2 50 HPLC Column HPLC Column HPLC Guard Column Guard Column Format Format HPLC Column HPLC Guard Column Format 3,4 Seconds 5 100 250 250 50 50 Length (mm) Length (mm) Length 250 50 Length (mm) Length 6 3828 150 063448 063464 063454 063470 2.0mm ID 2.0mm ID 2.0mm 039658 039657 4.0mm ID 4.0mm Peaks: 1. Horseheart Myoglobin 33μg Myoglobin 1. Horseheart 280nm UV, 5mL/min NaCl 350mM 10 to Peaks: Detection: Rate: Flow Eluent: ×50mm) (4 PA1 ProPac Column: 6. Soybean Trysin Inhibitor 66 Inhibitor Trysin Soybean 6. 66 Ovalbumin 5. 66 Conalbumin 3,4. – Contaminant 2. 8.2 pH Tris, 1.0mM in 054997 054999 054998 055150 4.0mm ID 4.0mm ID 4.0mm 040137 – 9.0mm ID9.0mm 063703 063707 – – 9.0mm ID9.0mm ID9.0mm SP5594 SP5598 – – 22.0mm ID 22.0mm ID 22.0mm ProPac HIC-10 LC Columns and Accessories Hydrophobic Interaction Chromatography columns for the high resolution separation of proteins and peptides

• High-resolution HPLC separation of proteins, protein variants and peptides • Proteins are separated under non-denaturing conditions • High protein loading capacity for protein puri cation applications • Wide range of applications • Based on 5μm ultra high purity spherical silica gel particles with 300Å pores

The ProPac HIC-10 column is a high-resolution, high-capacity, silica-based HIC column that provides excellent separations of proteins and variants for analytical and preparative applications. ProPac HIC columns provide exceptional hydrolytic stability under the highly aqueous conditions used in HIC.

>> ProPac HIC-10 Ordering Guide Columns for Biomolecules Particle Size (µm) Format Length (mm) 2.1mm ID 4.6mm ID 7.8mm ID 5 HPLC Column 75 – – 063665 100 063653 063655 – 250 – 074197 –

Protein mixture

600 Column: 7.5 × 75mm (Competitors A and B); 7.8 × 75mm (ProPac HIC-10) Flow: 1.0mL/min 4 100 WVL: 214nm 2 5 Eluents: (A) 2 M (NH )2 SO in 0.1 M NaH PO (pH 7.0) mAU 1 4 4 2 4 3 (B) 0.1 M NaH2PO4 (pH 7.0) Sample: Mixture of proteins 0 (1mg/mL each nal after 1:1 dilution with eluent A) Competitor A Inj. Volume: 20μL Competitor B Order of elution: 1. Cytochrome c -300 2. Myoglobin 0 10 20 30 40 50 60 70 3. Ribonuclease A Minutes 21286 4. Lysozyme 5. Chymotrypsinogen

Monoclonal antibody

1000 ® 10% CH3CN in eluent B Column: ProPac HIC-10, 4.6 × 100mm 100 Eluents: (A) 0.5 M (NH4)2 SO4

in 0.1M NaH2PO4 (pH 7.0) (B) 0.1 M NaH PO (pH 7.0) 70 2 4 Gradient: 70–100% B in 15 min Flow: 1mL/min mAU Inj. Volume: 5μL (25μg) Detection: UV, 214nm %B Sample: MAb 50μL (50mg/mL) + 450μL Eluent B 0 -100 0 2.0 4.0 6.0 8.0 10.0 12.2 Minutes 21538

www.thermoscienti c.com/chromatography 4-121 4 -122 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories and peptidesbyimmobilizedmetalaffinitychromatography The ProPacIMAC-10isahigh-resolutionanalyticalandsemipreparativecolumnforseparationofproteins • • • • • • semipreparative proteinandpeptideapplications Immobilized MetalAffinityChromatography(IMAC)columnforanalyticaland IMAC-10 ProPac -0.002 -0.007 Wide rangeofmetal-speci capplications High loadingcapacityforproteinpuri cationapplications Retention controlbyimidazoleconcentrationorpHgradient Resolve targetproteinsusingasinglecolumninhigh-resolutiongradientrun State-of-the-art technologyforreusablecolumnswithmetaltailoredspeci city High-purity separationsofmetal-bindingproteins 0.000 0.020 0.000 0.070 10 Particle Size (µm) Size Particle Guide Ordering IMAC-10 ProPac mAU mAU 01 02 30 25 20 15 10 5 0 0 Prion-related peptides Standard proteinswithsurface-exposedhistidines 1 repeat 1 1 2 01 20 15 10 5 3i 2 repeats 3 2 4 4i 5i 5 4 repeats 6i Minutes Minutes 6 7 3 HPLC Column HPLC Loading Column Format 53 540 35 30 25 20420 20416 7 6 5 4 3 2 1 Proteins withsurfaceexposedhistidine ioulaeAmyoglobin ribonuclease A 250 50 250 50 Length (mm) Length – 063617 – – 1.0mm ID 2.0mm ID 2.0mm 1.0mm ID X = sarcosine (N-methyglycine) X = sarcosine 7 6 5 4 3 2 1 0.5mL/min peptides Prion-derived Peak 280nm Wavelength: 15µL Rate: Flow Sample: %A Volume: Inj. t (min) =7.5 pH MNaCl, + 0.5 HEPES (E1) 20mM Gradient: Eluent: ×250mm) (4 IMAC-10 ProPac Column: 0.5mL/min A 280nm Wavelength: ribonuclease 1. 0.25mg/mL Rate: Flow 15µL Sample: %E1 Vol: Inject t (min) Gradient: =7.5 pH imidazole, E1 +100mM E2: =7.5 pH MNaCl, +0.5 MES E1: 20mM x250mm) (4mm IMAC-10 ProPac Column: 624063278 063274 063272 – – WGA(PHGGGWGA) WGQ(PHGGGWGQ) KKRPKPWGQ(PHGGGWGQ) KKRP(PHGXGWGQ) KKRPKP(PHGGGWGQ) PHGXGWGQ PHGGGWGQ # Sequence 0 7 0curve 95 30 10 0 =7.5 pH imidazole, E1 +50mM (E2) anhydrase carbonic 1.50mg/mL 3. 2. 1.00mg/mL myoglobin 0 7 0curve 96 40 15 0 063276 – 063667 4.0mm ID 4.0mm 4 2 4 2 4 063280 063615 063718 063710 9.0mm ID 22.0mm ID 22.0mm ID 9.0mm 063282 – – – ProSwift IEX LC Columns and Accessories Monolith IEX columns for high-resolution and fast protein analysis

ProSwift IEX monoliths provide excellent resolving power at high mass loading combined with fast analysis capability • High resolution • High loading capacity • Fast analysis • Wide range of operational  ow rates • Excellent stability over a wide pH range • Outstanding reproducibility and ruggedness

ProSwift polymer monolith (poly(Meth)acrylate) media are uniquely suited for separation of proteins.

Each monolith is a single cylindrical, sponge-like polymer rod containing an uninterrupted, interconnected

network of flow-through channels of a specific pore size. These large channels combined with the >>

monolith’s nonporous surfaces result in fast mass-transfer, high-resolution, and fast protein separations. Columns for Biomolecules The unique globular morphology of the polymer medium provides its high capacity.

ProSwift Ion Exchange Monolith Ordering Guide Functional Group Length (mm) 1.0mm ID 4.6mm ID WAX-1S 50 066642 064294 WCX-1S 50 066643 064295 SAX-1S 50 068459 064293 SCX-1S 50 071977 066765

Dynamic protein loading capacity of ProSwift WCX-1S 1 × 50mm

2 Column: ProSwift WCX-1S, 1 × 50mm 3 Eluents: A. 10mM Sodium phosphate (pH 7.6) B. 1 M NaCl in eluent A Gradient: 0% B for 2 min, 0–85% B in 7.5 min, 1 85% B for 3 min Flow Rate: 0.2mL/min Inj. Volume: 1–24μL

mAU Temperature: 30°C 72µg Detection: UV, 280nm 48µg Sample: Protein mix, 1mg/mL each 24µg Peaks: 1. Ribonuclease A 12µg 6µg 2. Cytochrome C 3µg 3. Lysozyme

0 4 8 12 Minutes

Protein separation

Column: ProSwift WAX-1S, 1 × 50mm Eluents: A. 10mM Tris, pH 7.6 B. 1 M NaCl in 10mM Tris, pH 7.6 Gradient: 5 to 55% of B in 13 min, hold for 2 min Flow Rate: 0.2mL/min

mAU Temperature: 30°C Detection: UV, 280nm Inj. Volume: 1.3μL Sample: 1.25mg/mL E. coli protein

0 5 10 15 Minutes 24270

www.thermoscienti c.com/chromatography 4-123 4 -124 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories -300 600 -100 450 700 mAU 0 Competitive comparison 0 0 0 Proteins minimal lossofresolution. The channelsalsoproducelowbackpressure,allowingtheuseofhigherlinearvelocities with nonporous surfacesresultinfastmasstransferforhigh-resolutionandprotein separations. medium sizechannels(2H&4H)toverylargechannel(3U)sizes.These andthemonolith’s interconnected networkofflow-throughchannelsaspecificporesize;ranging fromsmallchannel(1S), separation ofproteins.Eachmonolithisasinglecylindricalpolymerrodcontaining anuninterrupted, ProSwift polymerreversed-phasemonolithmediaare(polyStyrene-co-DVB)uniquely suitedforthe • • • • • • • and analysisthananybeadbasedcolumnsavailable. Deliver theoutstandingresolvingpowerofnonporousanalyticalmediacombined withfasterseparations resolution atexceptionallyhighflowratesforfastproteinseparationsandanalysis Reversed-phase monolithcolumnsthatuniquelyprovidetheadvantagesofhigh RP ProSwift 8mL/min High loadingcapacity High stringentwashcompatible,forexample,1MNaOH Outstanding reproducibilityandruggedness Excellent stabilityoverawidepHrangeof1to14 High throughputandimprovedproductivity Highest operational owratesavailable High resolutionathighspeed RP-4H RP-4H RP-3U RP-2H RP-1S Functional Group Guide Ordering RP ProSwift 1 ProSwift RP-2H 4mL/min 2 3 2mL/min 250 50 50 50 50 (mm) Length 4 1 4 5 Minutes Minutes 1 2 6 3 1 4 1mL/min 7 066640 069477 – – – 1.0mm ID 2 mL/min 8 mL/min 5 2 8 3 9 Competitor A ProSwift RP-2H 4 – – 064298 064296 064297 ID 4.6mm 2 8 10 5 22904 22903 11 ek: .Rbnces 1.5mg/mL A 1. Ribonuclease of proteins ve Mixture 214nm UV, Peaks: Sample: Detection: 5μL Temperature: 8mL/min 2or 30°C Volume: Inj. 1-75% B in 2mL/min: 6 min Rate: Flow Gradient:

Eluents: (A) DI H DI (A) Eluents: ×50mm 4.6 RP-2H, ProSwift 1. Columns: H DI (A) 8mL/min or 1, 4, 2, Eluents: Rate: Flow ×50mm) (4.6 RP-2H ProSwift Column: 1.5mg/mL of proteins ve Mixture 5μL A 1. Ribonuclease Elution: of Order Volume: Inj. 214nm Sample: Detection: UV 1-75% 1mL/min: B in 12 min Gradient:

.Oabmn 1.5 Ovalbumin 5. 0.9 0.5 anhydrase Carbonic 4. BSA 3. C Cytochrome 2. 1. Bin min 1-75% 8mL/min: (B) DI H DI (B) 2. Competitor A, 4.6 × 100mm, 15μm ×100mm, 4.6 A, Competitor 2. .Oabmn 1.5 Ovalbumin 5. C0.5 0.9 anhydrase Carbonic 4. BSA 3. Cytochrome 2. min 1.5 Bin 1-75% 8mL/min: 3min Bin 1-75% 4mL/min: 6min Bin 1-75% 2mL/min: (B) DI H DI (B) 2 2 2 2 O/CH O/CH O/CH O/CH 3 3 3 3 CN (95:5; V/V) +0.1% TFA V/V) (95:5; CN CN (5:95; V/V) +0.1% TFA V/V) (5:95; CN CN (5:95; V/V) + 0.1% TFA V/V) (5:95; CN CN (95:5; V/V) +0.1% TFA V/V) (95:5; CN 1.5 1.5 ProSwift ConA-1S LC Columns and Accessories For the highly efficient enrichment and purification of Con A binding glycans and glycoconjugates

The ProSwift ConA-1S affinity monolith column is unsurpassed for fast, highly efficient enrichment and purification of Concanavalin A (Con A) binding glycans, glycopeptides, and glycoproteins. • Highly ef cient enrichment and puri cation through high-mannose-Con A binding • Highly puri ed glycan and glycoconjugate products • High capacity and ligand density • High sample recovery • Low elution volumes • Fast separations • HPLC compatible

• Reusable for over one hundred puri cations >> The HPLC compatible ProSwift ConA-1S affinity column maintains the specificity and selectivity for Con A binding glycans and glycoconjugates. The high capacity monolith (poly(meth)acrylate) and ligand density Columns for Biomolecules of the ProSwift ConA-1S column facilitates the highly efficient enrichment of samples and can purify up to 2mg of glycoproteins. The high peak efficiency monolith (poly(meth)acrylate) column produces sharp, narrow peaks resulting in low elution volumes of highly concentrated products. The column is reusable, and many enrichments and purifications are possible with minimal loss of capacity.

ProSwift ConA-1S Ordering Guide Functional Group Length (mm) 5.0mm ID ConA-1S 50 074148

Fast, highly efficient enrichment and purification of a group of fluorescently labeled glycans

Column: ProSwift ConA-1S (5 x 50mm) Flow rate: 1.0mL/min Eluent A: 50mM NaOAc, 0.2M NaCl, pH 5.3,

with 1mM CaCl2 and 1mM MnCl2 Eluent B: 100mM α-methyl mannoside in eluent A Gradient: 0% B for 2 min, 0-100% B in 0.5 min, 100% B for 3.5 min Temperature: 25°C mV Sample: 2-AB labeled serum protein N-glycans Detection: Fluorescence, EM at 420nm

0 1 2 3 4 5 6 Minutes 27456

Glycopeptide enrichment

Column: ProSwift ConA (5 × 50mm) Eluent A: 50mM NaOAc, 0.2 M NaCl, pH 5.3,

with 1mM CaCl2, 1mM MgCl2 Eluent B: 100mM α-methyl-mannopyranoside in eluent A Flow Rate: 0.5mL/min Temperature: 25°C Detection: UV 214nm

mAU Sample: Trypsin digested HRP peptides Inj. Volume: 40μL Gradient: 100% A from 0–5 min, 100% B from 6–15 min, 100% A from 16–25 min

0 4 8 12 16 Minutes 27452 www.thermoscienti c.com/chromatography 4-125 4 -126 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • consisting ofadivinylbenzenecoreandhydrophilicsurface,optimizedforaffinityseparation. Thermo Scientificbiocompatiblesystems.Thecolumnisbasedonanovelnon-porouspolymericresin MAbPac ProteinAcolumnformatallowsrapidautomationofloading,binding,elutionandcollectionusing throughput andaccurateanalysisthroughacombinationoflowbackpressurehighefficiency. The antibody (MAb)titeranalysisofsamplessuchasharvestcellcultures(HCC).ThisHPLCcolumnoffershigh The ThermoScientificMAbPacProteinAcolumnisanaffinitydesignedtoprovidefastmonoclonal Fast MAbtiteranalysis AColumn Protein MAbPac Designed foreaseofuseandautomation Excellent samplerecovery Rugged High ef ciencycolumn Particle Size (µm) Size Particle Guide AOrdering Protein MAbPac 12 mAu 1,000 2,000 3,000 4,000 5,200 Harvest cellculturetiteranalysis -200 0.0 Unbound 0.2 Format HPLC Column HPLC 0.4 0.6 IgG 0.8 Length (mm) Length 35 Minutes

1.0 Area,mAu.min 10 20 30 40 0 02 1.2 04 4.0 mm ID mm 4.0 082539 1.4 Amount rabbitIgG,μg 06 1.6 08 1.8 100 0 2.0 Sample: MAb B, 5mg/mL Harvest Cell Culture Cell Harvest 5mg/mL B, MAb nm 280 Sample: 10μL Injection volume: 0.60 Bfor 100% mins, 0.2 Bfor 0% Detection: Temperature: 30°C 150mM Phosphate, Sodium 50mM Gradient: 150mM Phosphate, Sodium 50mM B: Eluent 2mL/min A: Eluent Rate: Flow ×35mm) (4 A, Protein MAbPac Column: mins,0% B for 1.20 mins 1.20 Bfor mins,0% 2.5 pH acetonitrile, NaCl,5% 7.5 pH acetonitrile, 5% NaCl, MAbPac SCX-10 LC Columns and Accessories Strong cation exchange column designed specifically for the high-resolution, high efficiency analysis of monoclonal antibodies and associated variants

• Exceptionally high resolution for monoclonal antibody variants • High ef ciency • Ideal for characterization and quality control assessment of monoclonal antibodies • Unmatched column-to-column and lot-to-lot reproducibility • Hydrophobic interactions essentially eliminated • Ideal for stability studies • Meets the regulatory requirements for biopharmaceutical characterization The unique nonporous pellicular resin provides exceptionally high resolving power, permitting the separation of monoclonal antibody variants that differ by as little as one charged residue. Hydrophobic interactions

with the resin are essentially eliminated for very efficient peaks. >>

MAbPac SCX-10 Ordering Guide Columns for Biomolecules Particle Size (µm) Format Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 3 HPLC Column 50 – 077907 – 5 HPLC Column 50 – 078656 – 150 – 085198 – 250 – 078655 – 10 Guard Column 50 075749 074631 – HPLC Column 50 – 075603 – 150 – 075602 – 250 075604 074625 SP6866

High resolution separations of monoclonal antibody variants 6 Column: MAbPac SCX-10 (4 × 250mm) Eluents: A. 20mM MES (pH 5.6) + 60mM NaCl 8 B. 20mM MES (pH 5.6) + 300mM NaCl Gradient: 15–36% B in 50 min Flow Rate: 1mL/min 11 Temperature: 30°C mAU Inj. Volume: 10μL Detection: UV at 280nm 5 Sample: MAb B, 5mg/mL 4 7 9 Peaks 1–5: Acidic variants 10 2 3 Peaks 6, 8, 11: C-Terminal Lys variants 1 12 13 14 15 16 17 Peaks 12–17: Basic variants 0

0 10 20 30 40 50 58 Minutes 27562

Baseline resolution of C-terminal lysine variants of a monoclonal antibody

27.5 Column: MAbPac SCX-10 (4 × 250mm) 6 Eluents: A. 20mM MES (pH 5.6) + 60mM NaCl B. 20mM MES (pH 5.6) + 300mM NaCl Gradient: 15–36% B in 50 min Flow Rate: 1mL/min Temperature: 30°C Inj. Volume: 5μL

mAU 6 Detection: UV at 280nm 7 Samples: 1. MAb B, 900μg in 100μL (no carboxypeptidase) 4 5 8 2. MAb B, 900μg in 100μL + 2 9 1 3 carboxypeptidase, 50μg, 5 2 0 4 incubation at 37°C for 3 h 2 3 9 10 1 11 1 Both Chromatograms: Peaks 1–5: Acidic variants Sample 1: Peaks 6-8: C-Terminal lysine truncation 0 10 20 30 40 50 58 variants of main peak Minutes 27564 Peaks 9–11: C-Terminal lysine truncation variants of minor variant peak Sample 2: Peak 6 results from peaks 6, 7, and 8 after CBP treatment. Peak 9 results from peaks 9, 10, and 11 afer CBP treatment www.thermoscienti c.com/chromatography 4-127 4 -128 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Improved MAbresolution rates forfasterseparation and aresuitableforoperationupto7,000psi.Higherpressurecompatibilityofthecolumnhardwareallowsusehighflow take advantageofsmallerresinsizeaswelllongercolumnlengthtomaximizetheresolutionMAbvariantseparation, strong cation-exchangephasewithspeciallydevelopedbio-inertPEEKlinedstainlesssteelcolumnhardware.Thesecolumns Higher resolutionandthroughputofMAbchargevariantUHPLCseparationscanbeachievedusingthesmallparticleMAbPac • • • • BioRS, strongcationexchangecolumndesignedformonoclonalantibodies andassociatedvariants SCX-10MAbPac Columns RS Suitable foroperationupto7,000psi High pressurecompatibility Specially developedbio-inertPEEKlinedstainlesssteelcolumnhardware UHPLC, highthroughputanalysis

5 (µm) Size Particle Guide Ordering RS SCX-10 MAbPac mAU 10.0 20.0 30.0 4 0 0.0 . 0

0.0 4.0 8.0

Acidic variants 12.0 BioRS Column BioRS Format 16.0 2.00 Lysine variants 2.94 20.0 n.a. Minutes 2.44 Other basicvariants 24.0 3.55 n.a. 250 150 50 (mm) Length 28.0 32.0 velocity andsampleloadwasused. truncation peaks.Equivalentlinear Peak resolutionisshownforlysine A 36.0 B 082673 085209 082674 ID mm 4.6 40.0 44.0 46.0 B: B: ~8900Psi A: mL/min 1.5 NaCl mM +300 5.6 pH MES mM 20 backpressure: mM +60 5.6 pH MES mM 20 Column Rate: Flow 5mg/mL MAb B: Eluent A: Eluent ×250mm 4.6 15μL Injection Volume: Sample: Dimension: 5μm SCX, MAbPac Column: Gradient: 33-53% in 20 min 20 in 33-53% Gradient: min 30 Bin 33-53% Gradient: MAbPac SEC-1 LC Columns and Accessories A size exclusion chromatography (SEC) column specifically designed for the high resolution separation and characterization of monoclonal antibodies (MAb) and their aggregates

• Anaylsis of monoclonal antibodies (MAb) and their aggregates • Analysis of MAb Fab and Fc fragments. • Hydrophilic bonded layer for minimal nondesired interactions between the biomolecules and the stationary phase • Nonmetallic and Biocompatible PEEK housing eliminates metal contamination from the column hardware • Stable surface bonding leads to low column bleed and compatibility with MS, ELSD and Corona CAD detection • Reproducible and rugged • Superior performance for the analysis of monoclonal antibodies, even using high and low-salt concentrations

The stationary phase is packed in bio-inert PEEK format and is compatible with different eluent conditions containing both

low and high concentrations of salt in mobile phases, and mass spectrometry friendly volatile eluents. >> Columns for Biomolecules MAbPac SEC-1 Ordering Guide Particle Size (µm) Format Length (mm) 4.0mm ID 5 Guard Column 50 074697 HPLC Column 150 075592 300 074696

Monoclonal antibody aggregate separation

200 Column: MAbPac SEC-1, 5μm Dimension: 4.0 × 300mm (PEEK) MAb Monomer Mobile Phase: 0.3 M NaCl in 50mM phosphate buffer pH 6.8 Temperature: 30°C Flow Rate: 0.20mL/min Inj. Volume: 2μL Detection: UV, 280nm mAU Sample: MAb (10mg/mL)

Aggregates

0

0 5 10 15 20 25 Minutes 27614

www.thermoscienti c.com/chromatography 4-129 4 -130 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • Included inthekit: column. ThiskitisaconvenientstarterandcolumnreplacementforMAbanalysis projects. The MAbAnalysisIEXandSECColumnKitincludestwocolumns:anion-exchange (IEX)columnandasize-exclusion(SEC) Kit and Column SEC IEX Analysis MAb • • Included intheKit: screening oftwocolumnsfordeterminationthebestcolumntheirspecificmonoclonal antibodysample. analysis. ThiskitisaconvenientstarterforresearchersatthebeginningofMAb analysisprojects,andfacilitatesthe MAb ChargeVariant AnalysisIEXColumnKitincludestwoion-exchange(IEX)specialty columnsforMAbchargevariants MAb ChargeVariant AnalysisIEXColumnKit Kits and Analysis Characterization Monoclonal Antibody MAbPac SEC-1Analyticalcolumn,4×300mm(P/N074696),asize-exclusion column designedforseparatingmonoclonal MAbPac SCX-10Analyticalcolumn,4×250mm(P/N074625),astrongcation-exchange columndesignedspeci callyfor MAbPac SCX-10Analyticalcolumn,4×250mm(P/N074625),astrongcation-exchange columndesignedspeci callyfor ProPac WCX-10Analyticalcolumn,4×250mm(P/N054993),aweakcation-exchange column,theindustrystandardfor MAb Analysis IEX and SEC Column Kit Column SEC and IEX Analysis MAb Description Guide Ordering Kit Analysis MAb Kit Column IEX Analysis Variants Charge MAb Description Guide Ordering Kit Variants Charge MAb antibody (MAb)monomers,aggregates,andfragments. high-resolution, high-ef ciencyanalysisofmonoclonalantibodiesandassociated variants. high-resolution, high-ef ciencyanalysisofmonoclonalantibodiesandassociated variants high-resolution, high-ef ciencyanalysisofmonoclonalantibodiesandassociated variants 076197 No. Cat. 076196 No. Cat. Thermo Scienti c pH Gradient Buffers LC Columns and Accessories Simple method development for charge variant characterization

The Thermo Scientific pH gradient platform accelerates method development and facilitates method transfer to QA/QC for a wide range of protein and MAb charge variants through a generic LC-based approach to charge variant characterization. • Patented buffer formulations enable fast, robust and reproducible pH gradients that are simple to optimize and easily automated • Ready to use with existing LC columns and systems, without the need for time consuming mobile phase adjustments • Applicable to the majority of MAbs

Thermo Scienti c pH buffer concentrates used in the pH gradient platform can be purchased individually or as a pair, in quantities of 125mL or 250mL. For added convenience, the 125mL

buffers can also be bundled with columns in a number of speci cally precon gured kits

• The CX-1pH gradient starter kit contains 125mL each of buffers A and B, plus a MAbPac >>

SCX-10, 10μm, 4 × 250mm column Columns for Biomolecules • The CX-1 pH gradient high throughput kit contains 125mL each of buffers A and B, plus a MAbPac SCX-10, 5μm, 4 × 50mm column • The CX-1 pH gradient high resolution kit contains 125mL each of buffers A and B, plus a MAbPac SCX-10, 5μm, 4 × 250mm column For the ultimate  exibility, the precon gured kits are also available as platforms, including the pH Designer Software.The options are listed in the table below:

pH Gradient Buffers Ordering Guide Description Cat. No. CX-1 pH Gradient Buffer A (pH 5.6), 125mL 083273 CX-1 pH Gradient Buffer B (pH 10.2), 125mL 083275 CX-1 pH Gradient Buffer Kit (pH 5.6 to 10.2), 125mL 083274 CX-1 pH Gradient Buffer A (pH 5.6), 250mL 085346 CX-1 pH Gradient Buffer B (pH 10.2), 250mL 085348 CX-1 pH Gradient Buffer Kit (pH 5.6 to 10.2), 250mL 085349 CX-1 pH Gradient Starter Kit (pH 5.6 to 10.2), 125mL 083381 CX-1 pH Gradient High Throughput Kit (pH 5.6 to 10.2), 125mL 083378 CX-1 pH Gradient High Resolution Kit (pH 5.6 to 10.2), 125mL 083272 CX-1 pH Gradient Starter Platform (pH 5.6 to 10.2), 125mL 083380 CX-1 pH Gradient High Throughput Platform (pH 5.6 to 10.2), 125mL 083376 CX-1 pH Gradient High Resolution Platform (pH 5.6 to 10.2), 125mL 083270 pH Designer Software 085022

Optimiztion of MAb charge variant separation using a linear pH gradient: 25% B (pH 6.75) to 50% B (pH 7.9)

16.0 8.00 pH trace

7.75 ]

10.0 mAU 7.50

7.25 pH 5.0

Absorbance [ 7.00

-2.0 6.60 0 5 10 15 20 25 30 35 40 Minutes www.thermoscienti c.com/chromatography 4-131 4 -132 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Separation of2ABlabeledN-glycansfrombovinefetuinbycharge,sizeandpolarity and LC-MSmethods. analysis withuniqueselectivityforbiologicallyimportantglycans,eitherlabeled ornative,byLC-fluorescence separation ofglycansbycharge,sizeandpolarity. Itisdesignedforhigh-resolutionandhigh-throughput 7E5 Thermo Scientific • • • • • High-Resolution ColumnsforGlycanAnalysis GlycanPac Column AXH-1 Highchromatographicef ciencyandexcellentcolumnstability Compatiblewith uorescenceandMSdetectionmethods Usefulforbothhigh-resolutionglycanpro lecharacterizationandeasyquanti cation Excellentresolutionforbothnativeandlabeledglycans Uniqueglycanselectivitybasedoncharge,sizeandpolarity 0 3 1.9 Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering AXH-1 GlycanPac

Fluorescence Counts of glycansbasedoncharge 0 neutral 1 2 ™ GlycanPac 36 monosialylated HPLC Column HPLC (2/pk) Cartridges Guard UHPLC Column Format 4 10 5 7 ™ 8 AXH-1isahigh-performance,silica-basedHPLC columnforsimultaneous 9 10 11 disialylated 12 13 150 10 250 150 100 Length (mm) Length 14 Minutes 15 20 16 17 trisialylated 18 19 20 21 082470 846082475 082476 082521 082472 082473 2.1 mm ID 3.0 mm ID 4.6 mm ID mm 4.6 ID mm 3.0 2.1 ID mm tetrasialylated 22 23 30 24 pentasialylated 25 26 849082468 082469 – – – 074188 069707 069580 No. Cat. 40 Column: GlycanPac AXH-1 (1.9μm) AXH-1 GlycanPac Column: Sample: 2AB labeled N-glycan from bovine fetuin bovine from N-glycan labeled 2AB at Fluorescence 320/420nm Sample: 30 °C Detection: 0.4mL/min Pmoles 50 Temperature: Vol.: Injection Rate: Flow (100%) Acetonitrile A: 2.1 ×150mm Phase Mobile Dimension: 082474 – – – Time (min) -10 40 30 35 0 %C %B % A 50 60 78 78 70 C: Ammonium formate (100mM, pH =4.4) pH (100mM, formate Ammonium C: Water B: 20 20 20 020 20 20 30 10 2 2 (mL/min) Flow 0.4 0.4 0.4 0.4 0.4 Curve 5 5 5 5 5 GlycanPac AXR-1 LC Columns and Accessories Ultra-high resolution UHPLC columns for glycan analysis

• Excellent glycan selectivity based on hydrophobicity, charge, size, and isomerization • High resolution for isomeric glycans • Compatibility with  uorescence and MS detection methods • High column ef ciency and stability • Ideal tool for qualitative, quantitative and structural analysis of glycans

The GlycanPac AXR-1 column, based on 1.9µm, high-purity and spherical silica substrates, combines both weak anion-exchange (WAX) and reversed-phase (RP) retention mechanisms for optimal selectivity and ultra-high resolution for glycan separation. The WAX functionality separates glycans based on charge, and RP property facilitates high resolution for glycans of the same charge according to their hydrophobicity, branching and isomerization. As the result, the GlycanPac AXR-1 column provides unparalleled resolutions for complex charged glycans. >> Columns for Biomolecules

Separation of 2AB labeled N-glycans from bovine fetuin using GlycanPac AXR-1

No. of detected trisialylated Column: GlycanPac AXR-1, 1.9µm peaks: ~80 disialylated Dimension: 2.1 × 150mm 7E6 Mobile phase: A: acetonitrile B: D.I. water C: ammonium formate (100 mM, pH =4.4) Flow Rate: 0.4 mL/min Temperature: 40 °C Injection Vol.: 100 pmoles tetrasialylated Detection:  uorescence at 320/420nm Sample: 2AB labeled N-glycan from bovine fetuin monosialylated

Fluorescence Counts Time % A %B %C Curve 2AB (minTime) % A % B % C Curve -10(min) 0 95 5 5 0 0 95 5 5 neutral -10 0 95 5 5 1 0 95 5 5 0 0 95 5 5 30 1 74 25 5 651 20 0 50 3095 5 5 5 0102030405060 65 30 1 74 25 5 Minutes 65 20 50 30 5

GlycanPac AXR-1 Ordering Guide Particle Size (µm) Format Length (mm) 2.1 mm ID 1.9 UHPLC Column 150 088136 250 088135

www.thermoscienti c.com/chromatography 4-133 4 -134 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Oligonucleotides functionalized ThermoScientific The DNAPacPA100 isa13μmpellicular, nonporouspolymericresinwithboundquaternaryamine- resolution. Itisstableunderdenaturingconditions,rugged,reliable,andcanbereadilyscaledup. The DNAPacPA100 isahigh-resolutionanion-exchangecolumnthatprovidessingle-base • • • • • • and purificationofsyntheticoligonucleotides A stronganionexchangecolumndevelopedtoprovidehigh-resolutionanalysis PA100 DNAPac DNAPac PA100 canresolvefulllengthfromn–1,n+1,andotherfailuresequences. of thisresinresultinveryhigh-resolutionoligonucleotideseparations. Provides easyscale-upfrom2.0mmto22mmIDcolumn(>100x) Analyzes phosphorothioate-basedclinicalsamples Compatible withsolvent,highpHortemperatures Resolves oligonucleotideswithsecondarystructures Capable ofn,n-1resolutionforoligonucleotides High-resolution oligonucleotideseparations 13 Particle Size (µm) Size Particle Guide Ordering PA100 DNAPac -0.001 0.004

mAU 5 HPLC Column HPLC Guard Column Format 10 ™ Dionex Minutes 15 ™ 250 50 Length (mm) Length MicroBeads 20 ™ . The rapid mass-transport characteristics . Therapidmass-transportcharacteristics SP3686 SP4016 2.0mm ID 2.0mm 10954 25 Detection: UV, 260nm UV, 1.5mL/min NaCl 410-510mM Sample: pd(A) Detection: Rate: Flow Eluent: PA100 DNAPac Column: 043010 043018 4.0mm ID 4.0mm in 25mM Tris-Cl, pH 8.0 pH Tris-Cl, 25mM in 40–60 043011 SP4511 9.0mm ID9.0mm

SP2091 SP4513 22.0mm ID 22.0mm

>> LC Columns and Accessories and Columns LC Columns for Biomolecules for Columns LC Columns and Accessories DNAPac PA200 LC Columns and Accessories The DNAPac PA200 is a strong anion exchange column developed to provide best- resolution for analysis and purification of synthetic oligonucleotides

• Industry leading resolution for oligonucleotide separations • Achieve n, n-1 resolution for oligonucleotides • Resolve oligonucleotides with secondary structures • Assay phosphorothioate purity • Selectivity control with eluent pH, salt, and solvent • Resolve RNA with aberrant (2’, 5’) links from normal SS-RNA • Separate individual phosphorothioate diastereoisomers • HR/AM AXLC/MS via automated desalting

The DNAPac PA200 is packed with a pellicular anion-exchange resin composed of an 8 μm diameter nonporous >> polymeric substrate to which quaternary amine-functionalized Dionex MicroBeads are bound. The rapid mass >> Columns for Biomolecules transport characteristics of this resin result in high-resolution oligonucleotide separations. DNAPac PA200 can Columns for Biomolecules resolve full length from n−1, n+1, and other failure sequences not possible with other columns.

DNAPac PA200 Ordering Guide Particle Size (µm) Format Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID 8 Guard Column 50 063423 062998 063419 SP6731 HPLC Column 250 063425 063000 063421 SP6734

Target, failure and trityl-on oligonucleotides Separation of oligonucleotides by length

15 80 30.1% 87.5% Oligonucleotide length: Full-length 21 22 23 24 25 Trityl-off Dx98 Dx96 Dx94 Dx91 Dx90 Dx88 Dx86 Dx87 D84 D80 55.8% 260 260 mA mA Failure Full-length 1.25 M NaCl: sequences 22.8% Trityl-off Trityl-on 22.8%

0.33 M NaClO4: 22.8%

0 0 0 8 16 24 3 4 5 6 7 Minutes 21022 Minutes 21024

Column: DNAPac® PA200

Eluent: NaClO4, pH 6.5

with 20% CH3CN Flow Rate: 1.2mL/min Inj. Volume: 8µL Detection: UV, 260nm

www.thermoscienti c.com/chromatography 4-135 4 -136 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories mA • • • • • to resolvefulllengtholigonucleotidesfromn-1andn+1other failuresequences. columns offerexceptionalresolutionofoligonucleotides,includingisomerseparations; andareable unwanted interactionsofeluentsandanalyteswithmetals,whilemaintaining10,000 psistability. They These columnsarepackedinbioinertPEEK-linedstainlesssteel(SST)bodies,designed toprotectfrom performance. Thesmallerparticlesalsoallowshortercolumnstoprovidesignificantly higherthroughput. DNAPac PA200 RScolumnsarepackedwithsmaller, 4μmparticles,forimprovedresolutionandbetter UHPLC SolutionsforNucleicAcidAnalysis RS PA200 DNAPac 16 10,000 psistable Ruggedness consistentwiththeDNAPacPA200 columnline. throughput Improved Higher ef ciencytoimproveresolution Provide singlebaseresolutionofoligonucleotides 4 Particle Size (µm) Size Particle Guide Ordering RS PA200 DNAPac 0

260 Partial resolutionof46oligonucleotides 0

dA12 PdA12 BioRS column Format PdA13 PdA14 1 PdA15 PdA16 PdA17 PdA18 dA20 PdA19 dA21 PdA20 PdA21 dA22 PdA22 250 150 50 Length (mm) Length dA23 PdA23 dA24 PdA24 PdA25 PdA26 PdA27 PdA28 Minutes PdA29 PdA30 2

PdA40 PdA41 PdA42 PdA43 PdA44 082510 082509 082508 4.6 mm ID mm 4.6 PdA45 PdA46 PdA47 PdA48 PdA49 PdA50 PdA51 3 PdA52 PdA53 PdA54 PdA55 PdA56 PdA57 PdA58 PdA59 PdA60 4 Chromeleon 6.8. concentration. Programed in salt amaximum approaching 40–60 PdA12–30, changing gradient, asymptotically continuously 3 indicates **Curve volumes =column *CV 4CV* Bin μL 2.5 28–43% Sample: mL/min 1.30 Detection: MNaCl A+1.25 (μL): Inj. 8 pH Tris mM 20 Rate: Flow Temp: B: Eluent A Eluent: x50mm 4.6 RS, PA200 DNAPac Column: (2.56 min) curve 3** curve min) (2.56 °C 30

>> LC Columns and Accessories and Columns LC Columns for Biomolecules for Columns LC Columns and Accessories DNASwift SAX-1S LC Columns and Accessories A strong anion exchange monolith column that provides improved capacity and industry-leading oligonucleotide yield-purity performance.

This semipreparative column incorporates selectivity control of the DNAPac column, providing unsurpassed purity and yields. • Micromole puri cations in a 5cm column body • Substantial capacity in a small format • Tunable selectivity control, the DNAPac columns, for high resolution • Compatible with high pH mobile phases, solvents, or high temperatures • Ideal for therapeutic and diagnostic oligonucleotide research • Purify dif cult oligonucleotide products • Use in combination with the DNAPac for industry leading puri cation and characterisation

of oligonucleotide products

>> The DNASwift column is a unique porous polymer monolith coated with functionalized latex >>

Columns for Biomolecules nanobeads, optimized for oligonucleotide separations. The monolith, a polymer cylinder with Columns for Biomolecules interconnected  ow through channels, provides fast mass transfer, low back pressure (for increased  ow rates), very high capacity, and re ned selectivity control.

DNASwift SAX-1S Ordering Guide Functional Group Length (mm) 5.0mm ID SAX-1S 150 066766

Tritylated oligonucleotide

Mixed-Base Tritylated 2800 20-mer Mixed-Base Column: DNASwift SAX-1S, 5 × 150mm 20-mer Mobile Phases: A: 15mM Tris, pH 8 B: 15mM Tris, pH 8, 1.25 M NaCl Gradient: 8–64% B in 10 min Flow Rate: 1.5mL/min Inj. Volume: 20µL mAU Detection: UV at 260nm Prep Cell (2mm path length) Sample: Derivatized mixed-base 20mer, 20mg/mL

0 0 4 8 12 Minutes 27019

Purification of a 21-base RNA Sample with aberrant 2' -5" linkages at the 1 and 3 positions from the 3" end

1200 Column: DNASwift SAX-1S, 5 × 150mm Eluents: A. 40mM Tris, pH 7 B. 40mM Tris, pH 7 + 1.25 M NaCl A Gradient: 26–42% B in 10 column volumes Dio-11: 5´-AUG AAC UUC AGG GUC AGC* UU*G-3´ Flow Rate: 1.5mL/min Inj. Amount: 125µg Temperature: 30°C mAU Detection: UV at 260nm

Fr 20 -200 0 5 10 15 20 Minutes

www.thermoscienti c.com/chromatography 4-137 4 -138 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Notes BioBasic SEC Columns LC Columns and Accessories Superior separation of water soluble compounds

• Covers separation of analytes over a wide molecular weight range • Long column life and high column ef ciencies • Simple mechanism of interaction based on molecular size and shape • Ideal for sample clean-up • Straightforward method development, simple mobile phases

BioBasic SEC Ordering Guide Pore Size (Å) Description ID (mm) Length (mm) Cat. No. BioBasic SEC 60 60 Guard 7.8 30 73305-037821 HPLC Column 7.8 150 73305-157846 HPLC Column 7.8 300 73305-307846 >>

BioBasic SEC 120 Columns for Biomolecules 120 Guard 7.8 30 73405-037821 HPLC Column 7.8 150 73405-157846 HPLC Column 7.8 300 73405-307846 BioBasic SEC 300 300 Guard 7.8 30 73505-037821 HPLC Column 7.8 150 73505-157846 HPLC Column 7.8 300 73505-307846 BioBasic SEC 1000 1000 Guard 7.8 30 73605-037821 HPLC Column 7.8 150 73605-157846 HPLC Column 7.8 300 73605-307846

1/2

4 Columns: BioBasic SEC, 5µm, 300 x 7.8mm 3 60Å Eluent: 0.1M KH2PO4 pH 7 Flow: 1.0mL/min 1 Detector: UV at 254nm 2 Injection: 20µL 4 Sample: 1. Thyroglobulin (MW669,00) 3 2. Ovalbumin (MW 45,000) 120Å 3. Angiotensin II (MW 1,046) 4. PABA (Vm) (MW 137) 4

2 3 1 300Å

3/4

2 1 1000Å

0 2 4 6 8 10 MIN

Molecular weight calibration curves Effect of pore size on SEC resolution

www.thermoscienti c.com/chromatography 4-139 4 -140 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • BioBasic 4 • • • • Optimized fortheseparationofawiderangepeptides BioBasic 8 • • • • Outstanding separationofsmalltomediumpeptides BioBasic 18 Excellent forLC-MSseparations Outstanding reproducibility, ef ciencyandcolumnlifetime Lower carbonloadforoptimalretentionoflargerpeptidesandproteins Based on300Åsilicaforoutstandingbiomoleculeperformance Excellent forLC-MSseparations Outstanding reproducibility, ef ciencyandcolumnlifetime An excellentstartingcolumnforproteinandpeptideanalysis 300Å poresizeforimprovedbiomoleculeanalysis Excellent forLC-MSseparations Outstanding reproducibility, ef ciencyandcolumnlifetime High peakcapacitystationaryphase 300Å poresizeformaximumperformancewithbiomolecules Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 5µm 5µ 5µm, 10µm mL 9 2 to9 9 9 USP L26 High 300 7 L1 25 14 BioBasic AX LC Columns and Accessories Optimized for the separation of proteins, peptides, other anionic species and polar molecules

• Weak anion exchange phase for multiple charged species • 300Å pore size for enhanced protein and peptide separations • Suitable for HILIC retention and separation of highly polar molecules • Superb stability under demanding pH conditions 5µmN/A

BioBasic SCX For the separation of proteins, peptides, and other cationic species

• Strong cation exchange phase based on sulfonic acid chemistry >> • Separation and retention of basic and other cationic species Columns for Biomolecules • 300Å pore size for enhanced protein and peptide separations • Outstanding stability under demanding pH conditions

5µm L52

BioBasic Ordering Guide Particle Length Format ID (mm) C18 C8 C4 AX SCX Size (µm) (mm) 5 Drop-in Guard 10 1.0 72105-011001 72205-011001 72305-011001 73105-011001 73205-011001 (4/pk) 2.1 72105-012101 72205-012101 72305-012101 73105-012101 73205-012101 3.0 72105-013001 72205-013001 72305-013001 73105-013001 73205-013001 4.0/4.6 72105-014001 72205-014001 72305-014001 73105-014001 73205-014001 HPLC Column 50 1.0 72105-051030 72205-051030 72305-051030 73105-051030 73205-051030 2.1 72105-052130 72205-052130 72305-052130 73105-052130 73205-052130 3.0 – – – 73105-053030 73205-053030 4.6 – – – 73105-054630 73205-054630 100 1.0 72105-101030 72205-101030 72305-101030 73105-101030 73205-101030 2.1 72105-102130 72205-102130 72305-102130 73105-102130 73205-102130 3.0 72105-103030 72205-103030 72305-103030 73105-103030 73205-103030 4.6 72105-104630 72205-104630 72305-104630 73105-104630 73205-104630 150 1.0 72105-151030 72205-151030 72305-151030 73105-151030 73205-151030 2.1 72105-152130 72205-152130 72305-152130 73105-152130 73205-152130 3.0 – – – 73105-153030 73205-153030 4.6 72105-154630 72205-154630 72305-154630 73105-154630 73205-154630 250 1.0 72105-251030 72205-251030 72305-251030 73105-251030 73205-251030 2.1 72105-252130 72205-252130 72305-252130 73105-252130 73205-252130 4.6 72105-254630 72205-254630 72305-254630 73105-254630 73205-254630

Format Length (mm) ID (mm) Cat. No. UNIGUARD Guard 10 1.0 851-00 Cartridge Holder 2.1 852-00 3.0 852-00 4.0/4.6 850-00

www.thermoscienti c.com/chromatography 4-141 4 -142 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 0 0 Budesonide andrelatedsubstances • • • • • • • High-resolution reversed-phaseseparationofproteinsandpeptides C18 Acclaim 300 Ultrapure silicawithlowmetalcontentandexhaustivebondingendcapping compatible LC-MS Reproducible fordependableresults Application testedforsuitabilityinpeptidemapping High-performance bondingchemistryon300Åporesilica High-ef ciency3μmsphericalsilicasubstrate Designed forhigh-resolutionpeptidemappingandproteinseparations 5 3 Guard to Analytical Column Coupler V-2 Coupler Column Analytical to Guard V-2 coupler) and (Holder Kit Guard Acclaim V-2 Holder Cartridge Guard SST Acclaim Description Guide Ordering Holder Guard Acclaim (µm) Size Particle Guide Ordering C18 300 Acclaim 0–400 mAUFS 0–24 mAUFS 5 1 2 (2/pk) Cartridges Guard HPLC Column HPLC Format 3 4 Minutes 10 7 5 6 10 150 50 Length (mm) Length 9 15 8 0 1 21523 20 069690 060264 060263 2.1mm ID 2.1mm Peaks: 7, 8. Budesonide epimers, 99% epimers, 7, Budesonide 8. 2001 Analy. Biomed. Pharm. J. PR; Byron M, Hindle S, Hou Reference: 500μg/mL Budesonide, 240nm at UV 340U; UVD Peaks: ASI-100 autosampler, 15μL Sample: Detector: Injection: Temperature: 30°C 1.0mL/min Flow: 15:1 Acetonitrile:ethanol (A) Phase: Mobile Pump: x150mm 4.6 3.0µm, C18 300 Acclaim Column: after three days three after B 66% Isocratic acid 0.1%(B) phosphoric HPG/4 P580 Summit 075721 063684 – 3.0mm ID 074188 069707 069580 No. Cat. 24:371-80. Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 069697 060266 060265 4.6mm ID 4.6mm 3µm 8 2.5 to 7.5 to 2.5 USP High 300 L1 25 14 Accucore Columns for Biomolecules LC Columns and Accessories Ultimate Core Performance for Biomolecule Separations

The range of Accucore HPLC columns packed with 150Å pore diameter particles allow biomolecule separations to benefit from the superb resolution and high speed enabled by the Core Enhanced Technology™ that Accucore columns are based on. The 150Å pore diameter solid core particles used in the Accucore 150-C18, 150-C4 and 150-Amide-HILIC columns are designed specifically to provide the optimum combination of retention and resolution for biomolecules. The chromatograms below show the improved resolution and peak shapes achieved with Accucore 150-C18 compared to a C18 solid core phase with an 80Å pore diameter.

>> Columns for Biomolecules

1.12 Accucore 150-C18

8.74 3.76 7.09 8.91 13.46 11.20

1.01 Conventional Solid Core C18

8.52

3.60 6.98 13.13

10.84

Improved resolution Improved peak shape

150 140 130 120 110 100 Peak Capacity Comparison 90

Peak Capacity Sharper peaks result in a significantly higher 80 peak capacity for 150-C18 compared to the 70 60 conventional solid core C18 column. Conventional Accucore 150-C18 Solid Core C18 Stationary Phase

Accucore 2.6µm 150Å HPLC and nanoLC Columns Optimum Conditions and Ratings Column ID Optimum Flow Rate Backpressure Rating Temperature Rating 75µm 300nL/min 800 bar 70°C 2.1mm 400µL/min 1000 bar 70°C 3.0mm 800µL/min 1000 bar 70°C 4.6mm 1800µL/min 1000 bar 70°C www.thermoscienti c.com/chromatography 4-143 4 -144 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Peptide separations column exhibitsexcellentretentiontimereproducibility. Precision ofretentiontimesiscriticalforreliableanalysis.TheAccucore150-C18 than acolumnpackedwith<2μmwideporefullyporousC18. provides muchnarrowerpeakwidths,thereforesignificantlyhighercapacity Higher peakcapacitiesfacilitateincreasedpeptideidentifications.Accucore150-C18 • • • Accucore 150-C18

mAU 150Åporediametermaterial Outstandingresolution Designedfortheseparationofpeptides 100000 120000 140000 160000 180000 200000 220000 240000 -40000 -20000 20000 40000 60000 80000 0 0 1 1 2 3 2 4 5 6 Minutes 7 3 8 4 9 5 10 11 6 12 13 7 14 15 1. Glycine-Tyrosine (220nm) UV 5µL Peaks: Detection: 35°C Injection: Temperature: 2min; for hold 15 min; Bover 0-50% Flow: acetonitrile:water 70:30 in 0.1% TFA Gradient: acetonitrile:water 10:90 in 0.1% TFA B: phase Mobile A: phase Mobile x2.1mm 100 2.6µm, 150-C18 Accucore Column: 7. Insulin A Ribonuclease 6. 5. Leu-Enkephalin III Angiotensin 4. 3. Met-Enkephalin 2 . Valine-Tyrosine-Valine 300µL/min 5min for B 0% at hold 0.1 min; in 0% to drop Particle Size Carbon Load(%) Pore Size pH Range Hydrophobicity 0 0 0 Low 2.6µm 7 1 to11 150Å USP 300 L1 25 14 LC Columns and Accessories Accucore 150-C4 Hydrophobicity • Signi cantly lower hydrophobic retention than C18 Low pH Range • Ideal for retention of proteins and larger peptides 0 2 to 9 14 Pore Size Accucore 150-C4 provides significantly sharper and higher peaks than a column packed with 5μm wide pore fully porous C4, thus offering better resolution and 0 150Å 300 sensitivity. The Accucore 150-C4 also performs better than a column packed with Carbon Load (%) <2μm wide pore fully porous C4 and generates only a fraction of the backpressure. 0 2 25 Particle Size 2.6µm USP L26 Intact proteins

Column: Accucore 150-C4 2.6µm, 100 x 2.1mm 130000 Mobile phase A: 0.1% TFA in 30:70 acetonitrile:water 120000 Mobile phase B: 0.1% TFA in 98:2 acetonitrile:water Gradient: 0-30% B over 8 mins 110000 5 30-95% B over 2 mins

95% B hold for 1 min 100000 0% B hold for 4 mins Flow: 400µL/min >> 90000 Temperature: 40°C Columns for Biomolecules 80000 3 Injection: 2µL 10pmol/µL solution Detection: UV (214 and 280nm) 70000 Peaks: 1. Insulin 2. Cytochrome C 60000 3. Lysozyme mAU 4. Myoglobin 50000 2 6 5. Carbonic anhydrase 40000 6. Ovalbumin 1 Carbonic anhydrase impurity 30000 4 * 20000 * 10000

0

-10000

012 3 4 5 6 7 8 9 10 Minutes

www.thermoscienti c.com/chromatography 4-145 4 -146 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories is anexcellentchoiceforglycanseparations. including carbohydratesandpeptides.AsaresulttheAccucore150-Amide-HILIC 150-Amide-HILIC iswellsuitedforseparatingavarietyofhydrophilicmolecules, HILIC phases.Combinedwithlargerporesizeofthesolidcoreparticles,Accucore stationary phaseandtheanalytes,resultinginuniqueselectivitycomparedtoother The amidebondedphasesprovidestronghydrogenbondinginteractionbetween the • • • Accucore 150-Amide-HILIC mAU Counts Recommendedforhydrophilicbiomoleculessuchasglycans HighretentionofabroadrangehydrophilicanalytesinHILICmode Amidephasebondedonto150Åporediametersolidcoreparticles 10 11 12 1000000 1100000 1200000 -200000 -100000 A further10glycansweredetected. (B) 5µLinjectionofsample,zoomed-intothelaterpartgradientrise. (A) 2µLinjectionofsample,where11glycanswereseparated. Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 0 1 2 3 4 5 6 7 8 9 100000 200000 300000 400000 500000 600000 700000 800000 900000 1 0 2-AB labelleddextranladder 2-AB labelleddextranladder 530 25 1 5 2.6µm 2 2 10 2 to8 3 3 15 AB 4 150 Å 54 45 40 35 4 5 20 100000 150000 200000 300000 350000 250000 50000 Minutes 6 5 0 Minutes 10 7 30 25 6 11 8 7 12 9 13 8 30 10 14 35 9 11 Minutes 15 10 16 50 35 11 High 300 17 14 12 18 13 40 19 14 20 40 55 15 21 16 17 18 45 45

solvent: 80:20 (v/v) acetonitrile:water. (v/v) 80:20 420nm wavelength; excitation 330nm parameters: acquisition sample. of 5μL to 2μL detector Fluorescence solvent: wash minutes 50 Injection 60 °C Injection: Temperature: time: Run conditions:110starting bar at Backpressure Flow: minutes 40.0 %Bin 20–50 4.5 pH formate, ammonium mM 50 Gradient: B: phase acetonitrile Mobile A: phase Mobile x2.1mm 100 2.6μm, Column: Accucore 150-Amide-HILIC, Detection: UV (240 and 33 nm) 33 and (240 UV bar 280 Detection: at 1μL μL/min 20 at 0.5μL Loading: Sample A) (100% bar 60 Pick-up: Sample 40 °C Backpressure: minutes 50 in %B 0–50 Temperature: Flow: water acetonitrile: (v/v) 2:98 Gradient: water acetonitrile: B: (v/v) phase 98:2 Mobile A: phase Mobile x 75μm 150mm 2.6μm, Column: Accucore 150-Amide-HILIC

500 μL/min 500 minutes 4.5 %Bfor 50 minutes 0.5 %Bin 50–20 minutes 5.0 %Bfor 50 200 nL/min 200 8minutes %Bfor 50 after 3 min from gradient start. gradient from 3min after start acquisition wavelength; emission

Counts 1000000 1100000 1200000

-200000 -100000 A further10glycansweredetected. (B) 5µLinjectionofsample,zoomed-intothelaterpartgradientrise. (A) 2µLinjectionofsample,where11glycanswereseparated. Particle Size Pore Size pH Range Hydrophobicity 0 0 Low 100000 200000 300000 400000 500000 600000 700000 800000 900000

0

1 5

2.6µm 2 10 2 to8 3

15 AB 4 150 Å 5 20 100000 150000 200000 300000 350000 250000 50000 6 35 0 Minutes 10 37 7 30 25 11 8 12 9 13 30 10 14 35 11 Minutes 15 16 35 High 300 17 14 18 40 19 20 40 21 45 45

35 Accucore Columns for Biomolecules Ordering Guide LC Columns and Accessories

Particle Size (µm) Format Length (mm) ID (mm) 150-C18 150-C4 150-Amide-HILIC 2.6 Defender Guard 10 2.1 16126-012105 16526-012105 16726-012105 (4/pk) 3.0 16126-013005 16526-013005 16726-013005 4.6 16126-014005 16526-014005 16726-014005 HPLC Column 30 2.1 16126-032130 16526-032130 – 3.0 16126-033030 16526-033030 – 4.6 16126-034630 16526-034630 – 50 2.1 16126-052130 16526-052130 16726-052130 3.0 16126-053030 16526-053030 16726-053030 4.6 16126-054630 16526-054630 16726-054630 100 2.1 16126-102130 16526-102130 16726-102130 3.0 16126-103030 16526-103030 16726-103030 4.6 16126-104630 16526-104630 16726-104630

150 2.1 16126-152130 16526-152130 16726-152130

3.0 16126-153030 16526-153030 16726-153030 >>

4.6 16126-154630 16526-154630 16726-154630 Columns for Biomolecules 250 2.1 – – 16726-252130

Format Length (mm) ID (mm) Cat. No. UNIGUARD Guard Cartridge Holder 10 2.1 852-00 3.0 852-00 4.6 850-00

www.thermoscienti c.com/chromatography 4-147 4 -148 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories The cartridgebasedtrapcolumnsare useat samples orwithhydrophilic peptides. for highresolutionseparationofrelatively clean with thetrapcolumnatend.These areideal the nanotrapdesign,featuresasinglecapillary Our trapcolumnscomeintwoformatsaswell, performance withimprovedeaseofuse. 1000 bar, offeringthesamelevels of are capableofwithstandingpressuresuptoa allowing tool-freenano-LCconnections.They the assemblyofPEEKsleeveconnections, nanoViper fingertightconnectionseliminate for ournano,capillary, andmicroLCcolumns. has nowbecomethestandardcolumnformat However mostimportantly, ThermoScientific sensitivity with50cmlongnanocolumns. short microcolumnsorhighresolutionand available toallowtuningforthroughputwith diameter. Inaddition,differentlengthsare capillary (300µm)andmicro(1000µm)internal We offermostchemistriesinnano (75µm), Our columnscomeinavarietyofformats. Formats with alowinternalvolume.Thiswillresult limited sampleamountsinanenvironment LC columnsisobtainedbyintroducingthe The (theoretical)sensitivityincreaseofthese in caseswherethesampleamountislimited. Nano, capillaryandmicrocolumnsareideal Introduction Nano, and Columns Capillary Micro LC Dionex Sensitivity ™ Nano Capillary Micro Narrowbore Conventional Name nanoViper ™ fingertightfittingsystem 75 300 1.0mm 2.1mm 4.6mm Diameter Internal µm µm ™

performance anddata. easier toachievestate-of-the-artnanospray Eliminating imperfectconnectionsmakesitfar incorporates ananoViperconnector. integrated inaready-madeassemblywhich high-voltage electrode,andemitterare which theseparationcolumn,columnheater, the useofspecificallydesignedcolumnsin minimize theimpactofdeadvolumesthrough Thermo ScientificEASY-Spray columns RSLCnano standardapplicationsmanual. more detailsontrapcolumns,pleasecheckthe sample qualitydemandsmorerobustness.For are idealforlargesampleamountsandwhen higher flowratesandhaveawiderID.These or facilitationofLC-MSinterfacing. columns arebeneficialine.g.metabolomics concentrating effectsofcapillaryandmicro LC columnsinproteomics,butcertainlythe The bestknownuseistheapplicationofnano internal diameter, asshowninthetablebelow. in asensitivitygainthatisdependentonthe chemistries forpeptidesandproteins. column portfoliohasastrongfocus on column reason thatournano,capillaryandmicro analysis ofpeptidesandproteins.Itisforthat are frombiologicaloriginandinvolvethe The mostcommon“samplelimited”sources Chemistries 3761 235 21 5 1 Sensitivity Theoretical Proteomics Peptide prefractionation Protein prefractionation LC-MS Metabolomics, DMPK, pharma molecules, Small application area Predominant dimension inamultidimensionalapproach. columns thatareapplicableasthefirst portfolio iscompletedbyaselectionof and peptideswithveryhighefficiency. Our that arecapableofseparatingbothproteins our rangealsoincludesmonolithiccolumns proteins. Inadditiontothesehallmarkcolumns accommodate awiderangeofpeptidesand available in100and300Åporesizeto capabilities. AcclaimPepMapcolumnsare heritage improvingpeptideseparation RSLC (C18,2µmparticle)rangebuildsonthis analysis formanyyears.TheAcclaimPepMap been thebenchmarkfornanoLCpeptide The AcclaimPepMaprange(C18,C8,C4)has Loadability Acclaim PepMap C18 100Å LC Columns and Accessories The standard for peptide separations in proteomics

• High resolution protein identi cation for biomarker discovery, veri cation or any other analysis • Highest sensitivity in LC-MS • Quality control by factory testing all columns

Acclaim PepMap and Acclaim PepMap RSLC columns are specially designed for high-resolution analyses of tryptic, natural, and synthetic peptides. The columns are often applied for LC-MS/MS peptide mapping for protein identification, biomarker discovery, and systems biology. Due to their high loading capacity, the columns are exceptionally suitable for the analysis of low abundant peptides in complex proteomics samples.

Protein mixture digest

Column: Acclaim PepMap C18 3µm

10.0 (75um ID x 50cm)

Flow Rate: 300 nL/min >> Mobile Phase: A: water + 0.1% formic acid 8.0

B: water/acetonitrile 20/80 v/v% Columns for Biomolecules + 0.08% formic acid Gradient: 4–55% B in 120min, 5 min wash 6.0 Detection: UV, 214nm Sample: Protein mixture digest Temperature: 35°C 4.0 mAU

2.0

0.0

-1.5 15 30 40 50 60 70 80 90 100 110 120 130 Minutes

Acclaim PepMap C8, 100Å An excellent alternative to Acclaim PepMap 100Å C18, when separating very hydrophobic peptides (e.g., non-tryptic peptides)

Acclaim PepMap C4, 300Å Ideal for the separation of larger hydrophobic peptides and proteins, providing higher recoveries

Acclaim PepMap C18, 300Å Wide pore version allows larger peptides and proteins

Accucore 150-C18 Designed to give narrow peaks and high peak capacity for peptide separations

Accucore 150-C4 Ideal for high resolution protein separations

Accucore 150-Amide-HILIC Well suited for separating a variety of hydrophilic molecules, including carbohydrates and peptides

www.thermoscienti c.com/chromatography 4-149 4 -150 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Separation Columns Separation 5 (µm) size Particle 5 (µm) size Particle 3 (µm) size Particle 5 3 2 (µm) size Particle Accucore 150-Amide-HILIC Ordering Guide Ordering 150-Amide-HILIC Accucore Guide Ordering 150-C4 Accucore Guide Ordering 150-C18 Accucore Guide ÅOrdering 300 C18 PepMap Acclaim Guide Ordering 300Å C4 PepMap Acclaim Guide Ordering 100Å C8 PepMap Acclaim Guide Ordering 100Å C18 PepMap Acclaim 2.6 2.6 2.6 Particle size (µm) size Particle (µm) size Particle (µm) size Particle 150 (mm) Length 150 (mm) Length 150 (mm) Length 250 150 50 150 50 150 50 (mm) Length 500 250 500 250 150 500 150 500 150 Length (mm) Length (mm) Length (mm) Length 50µm ID 75µm ID 300µm ID 300µm ID 75µm – ID 50µm ID 300µm ID 75µm – ID 50µm ID 300µm ID 75µm – ID 50µm – – – 164454 164537 164713 164560 164712 ID 300µm 164534 164563 164562 ID 75µm 164561 ID 50µm 164715 164714 164710 164709 677142 671163579 164708 164721 164720 164707 164723 164722 164706 – 164572 – – 164571 164568 164567 164570 164569 164540 164536 16726-157569 16526-507569 16526-157569 16126-507569 16126-157569 75µm ID 75µm ID 75µm ID 75µm nanoViper Column nanoViper Column nanoViper Column nanoViper nanoViper Column nanoViper Column nanoViper Column nanoViper Column nanoViper 164718 164900 – 164902 164901 164716 – – – – 164719 – 164899 164717 1000µm ID 1000µm ID 1000µm ID – – – – 164711 1000µm ID 163574 160326 160323 – – 75µm ID 75µm ID 75µm 161185 ID 75µm – 164261 160321 – – – ID 75µm Classic Column Classic Column Classic Column Classic Column Trap columns – nano trap design LC Columns and Accessories

Acclaim PepMap C18 100Å Ordering Guide Type Part number Particle Size ID Bed Length (mm) Total Length Quantity nanoViper Column 164535 3 75 20 150 2 164705 3 75 20 70 2 164564 5 100 20 150 2 Classic Column 164197 5 100 10 150 2 164199 5 100 20 150 2 164213 5 200 20 150 2 Note: 164705 is a shorter total length used for vented column set up for example with EASY nLC 1000

Trap Columns – Cartridge (Set of 5)

>> Acclaim PepMap C18 100Å Ordering Guide Columns for Biomolecules Length (mm) 300µm ID 1000µm ID 5 160454 160434 15 – 160438

Acclaim PepMap C8 100Å Ordering Guide Length (mm) 300µm ID 5 161194

Acclaim PepMap C4 300Å Ordering Guide Length (mm) 300µm ID 5 163591

Acclaim PepMap C18 300Å Ordering Guide Length (mm) 300µm ID 5 163589

Accucore 150-C18 Ordering Guide Length (mm) 300µm ID 5 16126-900379

Accucore 150-C4 Ordering Guide Length (mm) 300µm ID 5 16526-900379

Accucore 150-Amide-HILIC Ordering Guide Length (mm) 300µm ID 5 16726-900379

μ-Precolumn™ holder, 5mm, with 30μm ID connecting tubing, nanoViper ttings Length (mm) 300µm ID 5 164649

www.thermoscienti c.com/chromatography 4-151 4 -152 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories on 2Dseparations. RSLCnano standardapplicationsmanualontheThermoScientificwebsiteformore information A TiO2 trappingcolumncanbeaddedtotheapplicationforphosphopeptideanalysis. Checkthe analysis. TherearededicatedapplicationkitswhichutilizeeitherSCXorRPasthe firstdimension. Improving separationefficiencycanbeachievedbyaddingacolumntothesetup formultidimensional 1 x10 x10 st TiO2 RP SCX Chemistry 1 0 1 2 3 4 0 1 2 3 4 8 8 st dimension columns for 2D separations 2D columns dimension for Isolation oftwosynthethicphosphopeptidesfromaBSAtrypticdigest dimension columns Ordering Guide Ordering columns dimension Bound fraction Breakthrough ENIMRpSENSESQLTSK 1931.8 Da 164215 164205 164592 164566 164565 Number Part Minutes SVENLPEAGlpTHEQR 1758.8 Da 200µm ID x 1cm packed with TiO2 with x1cm ID packed 200µm TiO2 with x1cm ID packed 100µm column nanoViper PA2, Acclaim with packed x15cm, ID 300μm column nanoViper 300Å, 5μm, ASP, Polysulfoethyl with packed x15cm, ID 1.0mm column nanoViper 10 S, Poros with packed x10cm, ID 300μm Description 23731 A 45403530252015105 B Mobile Phases: (A) 0.05% TFA in DI H DI in TFA 0.05% (A) Phases: Mobile Gradient: 3–40% acetronitrile in 30 min 30 in acetronitrile 3–40% TiO Gradient:

Wash Solvent: 0.01% HFBA in DI H DI in HFBA 0.01% Solvent: Wash n.Vlm: 5μL Detection: MS 300nL/min 8μL/min Volume: Inj. Flow: Loading Rate: Flow Dimensions: 75μm x15cm 75μm H DI in HFBA 0.05% Solvent: Loading Dimensions: 3μm C18, PepMap Acclaim Column: Separation TiO with packed x1cm, ID 200μm Column: Trap 100μm ID x 1cm, packed with Acclaim PepMap C18, 5μm C18, PepMap Acclaim with packed x1cm, ID 100μm 2 Trap Eluent: 250mM NH 250mM Eluent: Trap DI H DI acetronitrile/ in TFA 0.04% (B) 2 O (80:20 v/v) O (80:20 4 HCO 3 in DI H DI in 2 2 O Phosphopeptide analysisPhosphopeptide analysisPhosphopeptide (6720.0335) (acidic) RP Nan x (basic) RP Cap LC, 2D off-line Automated (6720.0330) RP xnano SCX micro Peptides, of LC 2D off-line Automated (6720.0325) plugs Salt 2D Application O 2 O 2 O, pH 9.0 pH O, 2 , 5μm EASY-Spray Columns LC Columns and Accessories Nanoflow LC-MS utilizes very narrow fused silica columns and as a result, even the slightest inconsistency in column connection can result in leaks and dead volumes and are a frequent source of poor data, causing unstable spray and a significant loss in performance. Trouble-shooting connection issues can be difficult and time consuming, even for experts. Thermo Scientific™ EASY-Spray™ columns address the aforementioned issues by incorporating high-performance nanobore columns and integrating them within a radical new design. Packed with either C18 for peptide separations or with selected wide pore phases and monoliths for the separation of intact proteins, e.g. top-down proteomics or the structural identification of biotherapeutics, the “plug and spray” approach delivers state of the art performance in a simple to use format, making research routine.

Acclaim PepMap C18 100Å Ordering Guide Particle size (µm) Length (mm) 50µm ID 75µm ID

2 150 ES801 –

250 – ES802 >> 500 – ES803 Columns for Biomolecules 3 150 – ES800

Accucore C4 150Å Ordering Guide Particle size (µm) Length (mm) 75µm ID 2.6 150 – ES811

Acclaim PepMap C18 300Å Ordering Guide Particle size (µm) Length (mm) 75µm ID 5 150 – ES812

PepSwift Ordering Guide Length (mm) 200µm ID 250 – ES810

EASY-Spray Emitters EASY-Spray emitters are integrated devices consisting of a sprayer coupled with a transfer line. The EASY-Spray benefits of pre-made connections, as well as the easy installation in the MS ion source can now be exploited in cases where no chromatography is required, for example when tuning the mass spectrometer.

EASY-Spray Emitters Ordering Guide Description Cat. No. EASY-Spray emitter, nano ow (Emitter ID 7µm, Transfer line ID ES791 20µm, Transfer line length 500mm) EASY-Spray emitter, micro ow (Emitter ID 20µm, Transfer line ID ES792 75µm, Transfer line length 500mm)

www.thermoscienti c.com/chromatography 4-153 4 -154 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories of theseparationorimprovetrappingefficiency. Various ion-pairingagentscanbeusedintheloadingsolventand/ormobilephasestochangeselectivity and proteinswithoutnegativeimpactonthechromatographicperformanceorrecovery ofthecompounds. PepSwift Precolumnscanbeusedforpreconcentrationanddesaltingofsamples consistingofpeptides High-sensitivity proteomicsandbiotechapplicationsareeasilyperformedusingthese columns. microparticulate sorbents,providingimportantadvantagestothechromatographic separation. divinylbenzene copolymer, themonolithicstructureoffersahigh-qualityalternative totraditional analysis inproteinidentification,biomarkerdiscovery, andsystemsbiology. Basedonapolystyrene PepSwift andProSwiftmonolithiccolumnsarespeciallydesignedforfasthigh-resolution LC/MS • • • • • • • and Columns Micro HPLC Capillary (PS-DVB) and ProSwift PepSwift nanoViper ttingsforeasycolumninstallation lifetime Superior Wide rangeofcolumnIDsandlengthsavailable Highest column-to-columnreproducibility Highest sensitivityforLC/MS High-speed peptideandproteinseparations(<15min) High-resolution forproteinidenti cation,biomarkerdiscovery, andsystemsbiology 250 50 5 Length (mm) Length PepSwift and ProSwift, nanoViper Ordering Guide Ordering nanoViper ProSwift, and PepSwift 164543 164584 – 100µm ID 100µm 164542 164557 164558 200µm ID 200µm PepSwift – 164585 – 500µm ID 500µm – 164586 – 1000µm ID ProSwift EASY-Columns LC Columns and Accessories Excellence in nanoscale separations

• Compatible with any nanoscale HPLC system • Optimized for online LC-MS • Quality control on every column • Simple,  exible design

Using highly pure chromatographic media and biocompatible, metal-free fused silica capillaries, Thermo Scientific™ EASY-Column™ capillary LC columns are produced with a focus on design simplicity and strict quality control. As a result, EASY-Column capillary LC columns deliver outstanding chromatographic performance on any nano LC system.

EASY-Columns Ordering Guide

Description Quantity Cat. No. EASY-Column, 2cm, ID 100µm, 5µm, C18-A1 (Trap column) 3 SC001 >>

EASY-Column, 10cm, ID 75µm, 3µm, C18-A2 (Analytical column) 1 SC200 Columns for Biomolecules EASY-Column, 10cm, ID 75µm, 3µm, C18-A2 (Analytical column) 3 SC2003 HPLC-to-Column Connector kit Zero-dead-volume union 1 SC600 (1/32in OD tubing), 10x SC603 A/B mixing tee & Venting Tee for two-column setup Nanoliter-dead-volume tee 1 SC601 (1/32in OD tubing), 10 sleeves for 360µm OD fused silica (10xSC603). Connector Kit for two-column setup Zero-dead-volume union 1 SC602 (1/32in OD tubing), Nanoliter-dead-volume tee (1/32 inch OD tubing), 10x SC603 Sleeves (2cm, 1/32in OD) for 360µm OD fused silica 30 SC603

www.thermoscienti c.com/chromatography 4-155 4 -156 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories columns areavailable groups ontheanalyte ideal medium for theanalysisofcarbohydrates Thermo Scientific • • • Polymer-based columnsforcarbohydrateanalysis XP HyperREZ organic acidandsugaralcoholforms. XP columnsarealsoavailablein dedicated application areaorretention below tohelpchoosethebest tables the to Refer application requirements. counter-ion tomeetyour appropriate forms, enablingyoutochoosethe different counter-ion forms differences arisefromthe with thesulfonatedresin.Selectivity interactions withthemetalionassociated ligandoperation inadditiontothe exchange of mode exclusion of thegelprovidesasize lifetime. Controlofthedegree of cross-linking column increased for easily beregenerated analytes. Thecolumnscan eluting closely some and improvedresolutionof analysis faster can alsoberunatelevatedtemperatures,for dilute acidasamobilephase.Thecolumns they are stable at low pH,allowingtheuseof and organicacids.Unlikesilicabasedcolumns a 4 or 8% divinylbenzene an provide and content, a monodisperseresinwith on based are columns StableforlongcolumnlifetimesevenatlowpHandhightemperatures Ef cientandreproduciblemonodisperseparticles Designedforthedeterminationofcarbohydrates,saccharides, HyperREZ XP Carbohydrate Ca HyperREZ XP Sugar Alcohols Alcohols Sugar XP HyperREZ Acids Organic XP HyperREZ HyperREZ XP Carbohydrate Pb Carbohydrate XP HyperREZ HyperREZ XP Ca Type Column H Carbohydrate XP HyperREZ Phase HyperREZ XP Carbohydrate Na Carbohydrate XP HyperREZ HyperREZ XP Na XP HyperREZ H XP HyperREZ Pb XP HyperREZ organic acids,andalcohols ™ + HyperREZ 2+ 2+ + Application Areas Application Corn syrup Oligosaccharides, organic acids, glycoproteinconstituents, products fermentation wine fermentation, products, dairy Alcohols, monosaccharides juice, Fruit Anomer separation wine fermentation, products, dairy Alcohols, Adulteration of food & beverages, confectionary, disaccharides, food additives in H molecules. HyperREZXP interactions of the the interactions of + , Ca withthehydroxyl ™ times.HyperREZ XP Carbohydrate XPCarbohydrate column basedon + 2+ Counter-ion 2+ 2+ + , Pb Counter-ion Counter-ion Counter-ion 2+ , andNa

+

Acids column and alcohols,canbeseparatedusingaHyperREZXPOrganic Products offermentation,includingorganicacids,sugars 4MIN 0 8 8 8 10 8 Particle Size (µm) Size Particle 8 1 2 3 4 5 6 7 1. Maltotriose RI Sample: H 0.0001mM 0.7mL/min 57°C Detector: Temperature: Rate: Flow 69608-107780 Eluent: Number: Part x7.7mm 100 8µm, Acids, Organic XP HyperREZ 8 8% cross linkage cross 8% 8% cross linkage cross 8% linkage cross 8% linkage 4% cross 8% cross linkage cross 8% Porosity 8% cross linkage cross 8% REZ-9025 8. Ethanol 8. Acid 7. Acetic Glycerol 6. Acid Lactic 5. Acid Succinic 4. 3. Glucose 2. Maltose 2 SO 4 Na Analysis ofsportsdrinkusingaHyperREZXPCarbohydrate 0 + column 8µm, 10µm 0 0 30MIN 20 10 1 2 3 4 1. Dp8 RI Sample: 0.3mL/min 80°C Detector: Temperature: Rate: Flow let H 69310-307780 Eluent: Number: Part Na XP Carbohydrate HyperREZ 10µm, 300 x7.7mm 300 10µm, 5 6 7 8 9 9. Fucose 8. Dpl (Glucose) Dp2 7. Dp3 6. Dp4 5. Dp5 4. 3. Dp6 Dp7 2. 2 O REZ-9023 + , LC Columns and Accessories HyperREZ XP Ordering Guide Particle Size (µm) Format Length (mm) ID (mm) Cat. No. HyperREZ XP Carbohydrate H+ 8 Guard Cartridge (2/pk) 5 3.0 69008-903027 Guard Column 50 7.7 69008-057726 HPLC Column 300 7.7 69008-307780 HyperREZ XP Carbohydrate Ca2+ 8 Guard Cartridge (2/pk) 5 3.0 69208-903027 Guard Column 50 7.7 69208-057726 HPLC Column 300 7.7 69208-307780 HyperREZ XP Carbohydrate Pb2+ 8 Guard Cartridge (2/pk) 5 3.0 69108-903027 Guard Column 50 7.7 69108-057726 HPLC Column 300 7.7 69108-307780 HyperREZ XP Carbohydrate Na+ 10 Guard Cartridge (2/pk) 5 3.0 69310-903027 Guard Column 50 7.7 69310-057726 HPLC Column 300 7.7 69310-307780 HyperREZ XP Organic Acids 8 Guard Cartridge (2/pk) 5 3.0 69008-903027 Guard Column – – Inquire

HPLC Column 100 7.7 69608-107780

HyperREZ XP Sugar Alcohols 8 Guard Cartridge (2/pk) 5 3.0 69208-903027 >>

Guard Column – – Inquire Columns for Biomolecules HPLC Column 300 7.7 69708-254030

Format Length (mm) ID (mm) Cat. No. HyperRez Guard Cartridge Holder 5 3 60002-354

Retention Times of Common Saccharides (min) Conditions: Column: 300 x 7.7mm Saccharide H+ Ca2+ Pb2+ Mobile Phase: H2O Adonitol 11.5 14.9 20.4 Flow Rate: 0.6mL/min Detection: RI Arabinose 11.4 13.6 19.4 Temperature: 75°C (H+) Erythritol 12.7 15.6 20.3 85°C (Ca2+) 80°C (Pb2+) Fructose 10.6 13.5 19.3 Note: partial hydrolysis may occur with some saccharides using H+. Fucose 12.2 13.7 17.1 Galactose 1.07 12.2 15.6 Glucose 9.9 11.1 13.9 Glycerol 14.1 16.1 19.5 Lactose 8.6 9.7 12.8 Maltose 8.4 9.5 12.5 Maltotriose 7.7 8.7 11.9 Mannitol 11.0 17.3 28.9 Mannose 1.5 12.5 16.7 Raf nose 8.2 8.6 11.4 Sorbitol 11.1 20.7 N/A Sucrose 9.8 9.4 11.9 Xylose 10.6 12.0 15.0

www.thermoscienti c.com/chromatography 4-157 4 -158 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories system backpressures.ConnectingLCmodules, of variousdifferentconnectiongeometriesand improves chromatographic results, independent in liquidchromatography. TheVipersystem The ThermoScientific • • • • • • • UHPLC system and HPLC Provides easeofuseanddead-volumefreeplumbingeveryconventional Fittings Fingertight Viper Thermo Scienti c LCAccessories MP35N flexible stainlesssteel(SST)orbiocompatible HPLC andmodernUHPLCsystem.Together with dead-volume freeplumbingofeveryconventional fitting systemprovideseaseofuseandvirtually Fits narrowconnectionssuchas10-portvalvesandenablesmixedusewithdifferentdesigns Works withvirtuallyanyvalveandcolumnfrommanufacturer Easy touseduestainlesssteelorbiocompatibleMP35N Available indifferentinnerdiameters:0.1mm(0.004in),0.13mm(0.005in)or0.18mm(0.007in) Available indifferentlengths:65mmandfrom150to950mm100mmsteps Supports operatingpressuresupto1250bar(18,130psi) Provides virtuallyzero-deadvolume ngertightconnections 950 850 750 650 550 450 350 250 150 Union, SST 65 Plug, SST Plug, Description Viper Accessories (mm) Length MP35N Biocompatible and –SST Systems Fitting Fingertight Viper ngertightdesign ® capillaries, it opens a new dimension capillaries,itopensanewdimension ™ Dionex – – – 6040.2265 6040.2255 6040.2245 6040.2235 6040.2225 6040.2215 6040.2207 .m D S .m D P5 .3mI,ST01m D S 0.18mm ID, MP35N 0.18mm ID, SST 0.13mm ID, SST 0.1mm ID, MP35N 0.1mm ID, SST ™ Viper ™ fingertight fingertight The Viper fitting system overcomes these The Viperfittingsystemovercomesthese drawbacks whichcancompromiseefficiency. hand orusingtoolshaveconsiderable minimized. Conventionalfittingstightenedby LC systemandmustbe an of efficiency detrimental effectsontheseparation Extra columnvolumesinHPLChavethemost tools issimplewiththeVipersystem. valves, andcolumnsquicklyeasilywithout – – – – 6042.2360 6042.2350 6042.2340 6042.2330 6042.2320 6042.2306 6040.2304 6040.2303 Cat. No. Cat. ™ capillaries(1/32inOD)and 6040.2350 6040.2330 6040.2320 6040.2310 6040.2305 6040.2345 6040.2335 6040.2325 6040.2315 6040.2307 1 Each 1 Each Quantity 6040.2390 6040.2380 6040.2370 6040.2395 6040.2355 6040.2365 6040.2375 6040.2385 6040.2360 6040.2357 Scientific HPLC systemusers.AllThermo for columns and compatibility withvirtuallyalldifferent valves acceptable lifetime,anduniversal unifies robustperformance,easeofuse, connection tovirtuallyzero.TheVipersystem to reducethedeadvolumeofanyfluidic drawbacks by design, working withoutferrules fingertight fittingsystemasastandard. BioRS andSDsystemsareequippedwithViper ™ Dionex ™ UltiMate 6042.2375 – 6042.2355 – 6042.2365 6042.2337 6042.2327 6042.2315 – – ™ 3000 XRS, RS, 3000XRS,RS, LC Columns and Accessories

>> LC Accessories

Viper Fingertight Fittings Kits for UltiMate 3000 Systems

Viper Fingertight Fitting Kits for UltiMate 3000 systems Description SD Systems RS Systems Viper Capillary Kit for ISO, LPG or DGP pumps 6040.2302 6040.2301 Viper Capillary Kit for HPG pumps 6040.2309 6040.2308 Viper Capillary Kit for biocompatible RSLC systems – 6841.2301 On-line SPE Solution Kit for x2 Dual Standard or RSLC systems 6040.2802 6040.2801 Tandem Operation Solution Kit for x2 Dual Standard or RSLC systems 6040.2804 6040.2803 Application Switching Solution Kit for x2 Dual Standard or RSLC systems 6040.2806 6040.2805 Parallel Setup Solution Kit for x2 Dual Standard or RSLC systems 6040.2810 6040.2809 Inverse Gradient Kit for x2 Dual Standard or RSLC systems 6040.2819 6040.2820 Automated Method Scouting Solution Kit for Standard or RSLC systems 6040.2808 6040.2807 MS Connection Kit for MS-frontends with WPS autosampler, excluding UV detection 6720.0355 6720.0370 MS Connection Kit for MS-frontends with WPS autosampler, including UV detection 6720.0365 6720.0375 MS Connection Kit for MS-frontends with HPG-RS pump and OAS autosampler excluding UV detection – 6720.0372 MS Connection Kit for MS-frontends with HPG-RS pump and OAS autosampler including UV detection – 6720.0377

Description XRS Systems Viper Capillary Kit for standalone system with with WPS autosampler 6043.2301 Viper Capillary Kit for standalone system with with OAS autosampler 6845.2301A MS Connection Kit for MS-frontends with WPS autosampler excluding UV detection 6720.0380 MS Connection Kit for MS-frontends with WPS autosampler including UV detection 6720.0385 MS Connection Kit for MS-frontends with LPG-XRS pump and OAS autosampler excluding UV detection 6720.0372 MS Connection Kit for MS-frontends with LPG-XRS pump and OAS autosampler including UV detection 6720.0377 An overview of all UltiMate 3000 systems can be found in the Instruments spare parts section on 4-209.

www.thermoscienti c.com/chromatography 4-159 4 -160 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories below isabout20%increaseinpeakcapacity. their UHPLC,virtuallydead-volume-freecapabilities.Thetheoreticalperformance effectbetweenthetwoexamplechromatograms It sealsatthetipandnotwithaferrule.Nordoesit“grab”tubingsingle point.ItisthisfeaturethatgivesnanoViperfittings The nanoViperconnectionscomepreassembledandhighpressuretested.fittings donot“grab”thetubingatasinglepoint. of theconnection. fused silicatubingorcolumnatthepointwhereferruleiscompressed.The hastoholdthetubinginsleeveandseal potential deadvolume.Apartfromtheriskofvolume(anextremecaseindicated above),thereisalsoariskofdamagingthe sleeve, andfusedsilica,whichallneedtobeassembledcorrectly. Thesealingisattheferrule,whichtheoreticallyleavesalarge Classical connectionsinnanoLCinvolveeitherPEEKsil • • • • • • • The ThermoScientific Fittings Fingertight nanoViper as wellthemostexperiencedusers. by design.ItoffersnanoLCconnectionsthatneverfailandbringsa peaceofmindappreciatedbynovices, nanoViper ngertight ttingsallowyoutocon gureyournano,capillary, Paves thewayforeasy, tool-freenano,capillaryandmicroLCsetupofanyapplication Works withvirtuallyanyvalve,andcolumn,frommanufacturer Easy touse1/32"PEEKsheathedfusedsilicaand ngertightdesign Suitable fortemperaturesupto80°C Compatible withbackpressuresupto1000bar(14,500psi) Provides virtuallyzero-dead-volumeconnections and microLCapplicationcompletelytoolfree. ™ Dionex ™ nanoViper ™ fingertightUHPLCfittingsystemisvirtuallydeadvolume-free ™ orfusedsilicainPEEK ™ sleeves.Theseconnectionsconsistofanut,ferrule, LC Columns and Accessories nanoViper Fingertight Fittings Length (mm) Orange Brown Black Red Purple 20µm ID 50µm ID 75µm ID 100µm ID 150µm ID 70 6041.5120 6041.5123 6041.5126 6041.5820 6041.5817 150 6041.5121 6041.5124 6041.5127 6041.5811 6041.5818 250 – – 6041.5730 6041.5812 6041.5819 350 6041.5240 6041.5540 6041.5735 6041.5813 6041.5820 450 – – – 6041.5814 6041.5821 550 6041.5260 6041.5560 6041.5760 6041.5815 6041.5822 650 6041.5275 6041.5575 6041.5775 – – 750 6041.5280 6041.5580 6041.5780 6041.5816 6041.5823 950 6041.5122 6041.5125 6041.5128 – –

Trap Column Cartridges Holders with nanoViper Fittings Description Cartridge Length (mm) Fitting ID (µm) Cat. No. µ-Precolumn holder for trap cartridges with 2 x 100mm 5 30 164649

length nanoViper ttings 15 75 164650 >>

Sample Loops with nanoViper Fittings LC Accessories Volume (µL) Cat. No. Quantity 1 6826.2401 1 Each 5 6826.2405 1 Each 10 6826.2410 1 Each 20 6826.2420 1 Each 50 6826.2450 1 Each 125 6826.2412 1 Each nanoViper Application Kits nanoViper fittings are also included in tubing and application kits designed for use with the UltiMate 3000 RSLCnano systems.

nanoViper Fitting Kits for UltiMate 3000 RSLCnano Systems RSLCnano Description Tubing Samples Trap Column Separation Column Systems Direct Injection nano LC Kit YY– Y 6720.0300 Direct Injection capillary LC Kit YY– Y 6720.0305 Preconcentration nano LC kit YYY Y 6720.0310 Preconcentration capillary LC kit Y Y Y Y 6720.0315 Preconcentration monolithic LC kit YYY Y 6720.0320 2D salt plugs kit YYY Y 6720.0325 Automated off line SCX-RP peptides kit Y Y Y Y 6720.0330 Automated off line RP-RP peptides kit Y Y Y Y 6720.0340 Tandem nano LC kit YYY Y 6720.0335 FLM nanoViper tubing kit Y–– – 6041.5100 (All tubing for nano LC preconcentration) MS connection kit Y–– – 6720.0345 EASY-Spray connection kit YYY – 6720.0395 (Supports direct injection as well as preconcetration)

www.thermoscienti c.com/chromatography 4-161 4 -162 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • connections self-adjusting Universal ConnectorsSLIPFREE Fingertight connectionsto10,000psi threads Stainless-steel Compatible withallcolumnend- ttings Void-free andleak-freebypushingtubingferrule Double Double Connectors, SLIPFREE Single Flexible Single Single Connectors, SLIPFREE Double Long-neck Double PEEKCollared Double Flexible Single Long-neck Single PEEKCollared into theend- tting 15 10.5 50 30 20 10 6 (cm) Length 20 10 20 10 6 40 28 15 10.5 40 30 20 10 6 (cm) Length 20 10 20 10 6 60 50 40 28 SLIPFREE FerrulesSLIPFREE Kel-F Vespel PEEK Material 01-LX315FE – – 39115-FLEX 39111-FLEX 30115-FLEX 30111-FLEX 30150 30130 30120 30110 30106 31720 31710 – – 30420 – 30410 39240-FLEX – 30406 39228-FLEX 39215-FLEX 30240-FLEX 39211-FLEX 30228-FLEX 30215-FLEX 30211-FLEX 30240 30230 30220 30210 30206 30520 30510 30320 – 30310 – – 30306 – 39140-FLEX 30160-FLEX 39128-FLEX 30150-FLEX 30140-FLEX 30128-FLEX 0.005in ID 0.005in 0.005in ID 0.005in – 31130 31120 31110 31106 32720 32710 31420 31410 31406 – 31230 31220 31210 31206 31520 31510 31320 31310 31306 0.010in ID 0.010in 0.010in ID 0.010in 36025 36024 36023 No. Cat. – 32130 32120 32110 32106 – – 32420 32410 32406 – 32230 32220 32210 32206 32520 – 32320 32310 32306 – – 0.020in ID 0.020in 0.020in ID 0.020in 1 Each 1 Each 1 Each Quantity PTFE One-Piece Column Connector LC Columns and Accessories Excellent for high-throughput screening and quick connection

• Fingertight, leak-free connection of analytical and guard columns with 10-32 threads • Minimizes dead volume • Inert and biocompatible material

PEEK One-Piece Column Connector Description Cat. No. Quantity One Piece Coupler 60170-370 1 Each

Solvent Inlet Filters >>

Feature a large surface area for a long lifetime LC Accessories

• 10μm inlet lters for longer lifetime • No tools required for replacement

Solvent Inlet Filters for UltiMate 3000 Systems Description Model Cat. No. Quantity Filter frit, porosity 10µm, V4A UltiMate 3000 pumps 6268.0110 10 Each stainless steel Filter frit, porosity 10µm, titanium UltiMate 3000 pumps 6268.0111 10 Each Filter holder UltiMate 3000 pumps 6268.0115 6 Each Solvent supply line, 1.5 x 3.0 x UltiMate 3000 pumps 6030.2548 1 Each 1000mm

Bottom-of-the-Bottle solvent lters • Ef cient draw • 100% PTFE polymer, including 2µm lters • Built-in helium sparge port and frit Solvent Filters for HPLC Systems Type For Use with Cat. No. Quantity

Stainless Steel Fit 1/16in OD tube to 1/8in OD plastic tubing A-302 1 Each

Stainless Steel Fit to 1/8in OD plastic tubing using 1/8in PP nut A-302A 1 Each

Bottom-of-the-Bottle 3/16in OD plastic tubing A-436 1 Each

Bottom-of-the-Bottle 1/8in OD tubing A-437 1 Each

www.thermoscienti c.com/chromatography 4-163 4 -164 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • Connects 30to50mmIDpreparativecolumns1/16intubing Reducing Columns Union Preparative for • • Accommodate awiderangeofconfigurations and Ferrules Nuts Steel High Stainless Pressure Reducing Union for Preparative Column Reducing Union forPreparative Thermo Scienti c High Pressure Stainless Steel Nuts and Ferrules and Nuts Steel Stainless Scienti Thermo Pressure c High ihu frit Without bore 1.0mm Stainless steelconstruction Burr andcontaminantfree Designed for10-32portcon gurations 1/8in to 1/16in Reducing Union for Preparative Column Preparative for Union 1/16in to Reducing 1/8in Description Male hex nut, Waters compatible Waters nut, hex Male 0.625in ferrules, Valco thread 10-32 nut, hex male Valco 0.625in ferrules, Universal nut hex Male Ferrules PEEK Replacement ferrule with nut thread 10-32 Type 60182-357 Cat. No. Cat. U-410x U-321x U-320x U-401x U-400x F-192x F-190 No. Cat. 1 Each Quantity 10 Pack 10 Pack 10 Pack 10 Pack 10 Pack 10 Pack 10 1 Each Quantity PEEK Fingertight Fittings LC Columns and Accessories Machined for reliability and ease of use

• Resist cracking, breaking, thread stripping and leaking in both low and high pressure applications • Biocompatible for a broad range of applications

PEEK Fingertight Fittings Type Cat. No. Quantity One-piece Fingertight Fitting, 1/16in, 0.37in head F-120x 10 Pack

One-Piece Long Fingertight Fitting, 1/16in, 0.37in head F-130x 10 Pack

One-Piece PEEK Fingertight Fitting, 1/32in, 0.25in head M-645x 10 Pack

Two-Piece Fingertight Wing Nut with Ferrule, 1/16in F-300x 10 Pack

Replacement PEEK Ferrules F-142x 10 Pack

Column End Plugs, 1/16in, 10-32 coned, Delrin, Black U-467BLKx 10 Pack >>

Column End Plugs, 1/16in, 10-32 coned, Delrin, Red U-467Rx 10 Pack LC Accessories

Viper Unions Suitable only for direct connection of two Viper capillaries

Description Cat. No. Quantity Viper union, stainless steel 6040.2304 1 Each

Stainless Steel Unions, Tees and Crosses Well-suited to high pressure applications

• Absolute zero or low dead volume formats • Includes two stainless steel nuts and ferrules

Stainless Steel Unions, Tees and Crosses Description Through Hole (in) Swept Volume (µL) Cat. No. Quantity Union, stainless steel, Upchurch Scienti c/Parker ttings 0.010 0.025 U-435 1 Each compatible, includes 2 stainless steel nuts and ferrules

Union, stainless steel, Upchurch Scienti c/Parker ttings 0.020 0.134 U-402 1 Each compatible, includes 2 stainless steel nuts and ferrules

Union, stainless steel, Upchurch Scienti c/Parker ttings 0.050 0.836 U-437 1 Each compatible, includes 2 stainless steel nuts and ferrules

Union, stainless steel, Upchurch Scienti c/Parker ttings 0.062 ~0.0 U-438 1 Each compatible, includes 2 stainless steel nuts and ferrules

Union, stainless steel, Waters ttings compatible, 0.020 0.129 U-412 1 Each includes 2 stainless steel nuts and ferrules

Union, stainless steel, Valco ttings compatible, 0.020 0.103 U-322 1 Each includes 2 stainless steel nuts and ferrules

Tee, stainless steel, 10-32 ttings 0.020 0.57 U-428 1 Each for use with 1/16in OD tubing

Cross, stainless steel, 10-32 ttings 0.020 0.72 U-430 1 Each for use with 1/16in OD tubing

www.thermoscienti c.com/chromatography 4-165 4 -166 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • Well-suited tohighpressureapplications Tees Unions, PEEK and Crosses PEEK and PEEK Lined Unions, Tees Crosses and Unions, Lined PEEK and PEEK Biocompatible Absolute zeroorlowdeadvolumeformats Union, PEEK polymer, includes two PEEK 2-piece 2-piece PEEK ttings two includes polymer, PEEK Union, four 10-32 PEEK double-winged nuts double-winged PEEK 10-32 four includes 1/16in tubing, OD with 10-32 use for PEEK, ttings nuts double-winged PEEK 10-32 three includes 1/16in tubing, OD with 10-32 use for Tee, PEEK, ttings 2-piece PEEK ttings two includes polymer, PEEK Union, Description 0.020 0.020 0.020 0.010 (in) Hole Through 0.28 0.72 0.57 0.070 (µL) Volume Swept P-704 P-729 P-727 P-742 No. Cat. 1 Each 1 Each 1 Each 1 Each Quantity PEEKsil Capillary Tubing LC Columns and Accessories Excellent chemical compatibility and very low carryover

• Precision-bore fused silica tubing coated with 1/16in OD PEEK covering • Usable in most standard chromatography systems • Withstands high pressures • Smooth internal surface for excellent  ow characteristics • Tubing is stiff: not recommended for uses requiring tubing bends • Precut lengths only: cutting in the lab may damage tubing

PEEKsil Capillary Tubing ID (in) Length (cm) Cat. No. Quantity

0.002 10 60182-500 5 Pack Applications: >> 20 60182-501 5 Pack • HPLC

50 60182-502 2 Pack LC Accessories • LC-MS 0.004 10 60182-503 5 Pack 20 60182-504 5 Pack 50 60182-505 2 Pack 0.007 10 60182-506 5 Pack 20 60182-507 5 Pack 50 60182-508 2 Pack

PEEK Sleeves for Fused Silica Capillary Tubing Withstands high pressures

1/16in OD PEEK Sleeves for Fused Silica Capillary Tubing ID (in) Color Cat. No. Quantity 0.008 Yellow F-227 1 Each 0.010 Blue F-228 1 Each 0.012 Natural F-229 1 Each 0.015 Orange F-230 1 Each 0.021 Natural F-231 1 Each 0.030 Natural F-232 1 Each

www.thermoscienti c.com/chromatography 4-167 4 -168 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • • Cleaned, polished,passivatedandready-to-use Tubing Capillary Steel 316 Stainless 1/16in 316 Stainless Steel Tubing, 5-Foot Coil Tubing, 5-Foot Steel Stainless 316 1/16in Tubing Capillary Steel Stainless 316 Rough internalsurfacemayleadtosamplecarryover Not recommendedforbiologicalsamples Pre nished,square,burr-free endsandinteriorstominimizedeadvolumeconnections Wide chemicalcompatibilty Suitable forultrahighpressureapplications 0.007 0.005 Sleeves 1/16in with Tubing Precut OD 1/32in 0.010 0.007 0.005 Tubing Precut OD 1/16in 0.046 0.040 0.030 0.020 0.010 0.007 0.005 (in) ID (in) ID 40 28 15 10.5 40 28 15 10.5 100 50 30 20 10 5 100 50 30 20 10 5 100 50 30 20 10 5 Length (cm) Length Yellow Yellow Yellow Yellow Red Red Red Red Blue Blue Blue Blue Blue Blue Black Black Black Black Black Black Red Red Red Red Red Red Color U-151 U-144 U-107 U-105 U-106 U-108 U-158 No. Cat. 39040-FLEX 39028-FLEX 39015-FLEX 39011-FLEX 30040-FLEX 30028-FLEX 30015-FLEX 30011-FLEX U-133 U-132 U-114 U-113 U-112 U-111 U-131 U-130 U-129 U-128 U-127 U-126 U-157 U-156 U-155 U-154 U-153 U-152 Cat. No. Cat. 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity PEEK Capillary Tubing LC Columns and Accessories Pre-cut and color-coded for easy identification and use

• Broad chemical compatibility • Biocompatible • Easily cut to desired length • Appropriate for many HPLC applications • Resistant to most organic solvents, but nitric acid, sulfuric acid, dichloromethane, THF and DMSO are not recommended

1/16in OD Precut PEEK Tubing ID (in) Length (cm) Color Cat. No. Quantity 0.003 5 Natural 37003-5 1 Each 10 Natural 37003-10 1 Each

20 Natural 37003-20 1 Each

30 Natural 37003-30 1 Each >>

50 Natural 37003-50 1 Each LC Accessories 100 Natural 37003-100 1 Each 0.005 5 Red 37005-5 1 Each 10 Red 37005-10 1 Each 20 Red 37005-20 1 Each 30 Red 37005-30 1 Each 50 Red 37005-50 1 Each 100 Red 37005-100 1 Each 0.007 5 Yellow 37007-5 1 Each 10 Yellow 37007-10 1 Each 20 Yellow 37007-20 1 Each 30 Yellow 37007-30 1 Each 50 Yellow 37007-50 1 Each 100 Yellow 37007-100 1 Each 0.010 5 Blue 37010-5 1 Each 10 Blue 37010-10 1 Each 20 Blue 37010-20 1 Each 30 Blue 37010-30 1 Each 50 Blue 37010-50 1 Each 100 Blue 37010-100 1 Each 0.020 5 Orange 37020-5 1 Each 10 Orange 37020-10 1 Each 20 Orange 37020-20 1 Each 30 Orange 37020-30 1 Each 50 Orange 37020-50 1 Each 100 Orange 37020-100 1 Each

1/16in OD PEEK Tubing, 5-Foot Coil ID (in) Cat. No. Quantity 0.003 37003 1 Each 0.005 37005 1 Each 0.007 37007 1 Each 0.010 37010 1 Each 0.020 37020 1 Each 0.030 37030 1 Each 0.040 37040 1 Each

www.thermoscienti c.com/chromatography 4-169 4 -170 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Fused SilicaFused Cutter Produce clean,90°cutsofstainlesssteeltubing Terry Tool Tubing Cutters • • Produces aflat,90°,burr-free end Tubing Cutter Polymer Fused Silica Cutter Silica Fused ToolTerry Tubing Cutters Tubing Cutter Polymer Guide holesfor1/16inand1/8intubing Compatible withrigidpolymerictubing Cutter for fused silica capillaries silica fused for Cutter Description tubing steel stainless 1/8in tubing steel 1/16in stainless Description blades Replacement Tubing Cutter Polymeric Description 6720.0016 No. Cat. 60182-510 60182-509 No. Cat. A-328 A-327 No. Cat. 1 Each Quantity 1 Each 1 Each Quantity 5 Pack 1 Each Quantity Rheodyne 7725 and 7725i Sample Injectors LC Columns and Accessories Allow continuous flow between the load and inject positions to protect against pressure shock

• Stainless steel construction • Make-Before-Break (MBB) design • Can use partial lling for zero sample waste or complete lling for better reproducibility • Inject 1µL to 5mL with high accuracy and precision • 7725i features a position sensing switch for a reproducible start signal

Rheodyne 7725 and 7725i Sample Injectors Model Mode Features Cat. No. Quantity 7725 Dual C o n t i n u o u s  o w 7725 1 Each 7725i Dual Continuous  ow, position sensing switch 7725i 1 Each

>> LC Accessories Rheodyne 9725 and 9725i Sample Injectors Allow continuous flow between the load and inject positions to protect against pressure shock

• Biocompatible PEEK construction • Make-Before-Break (MBB) design • Can use partial lling for zero sample waste or complete lling for better reproducibility • Inject 1µL to 5mL with high accuracy and precision • 9725i features a position sensing switch for a reproducible start signal

Rheodyne 9725 and 9725i Sample Injectors Model Mode Features Cat. No. Quantity 9725 Dual C o n t i n u o u s  o w 9725 1 Each 9725i Dual Continuous  ow, position sensing switch 9725i 1 Each

Rheodyne 8125 Low-dispersion Microscale Injector Designed for use with 1 and 2mm ID HPLC columns

• Can use partial lling for zero sample waste or complete lling for better reproducibility • Position sensing switch provides reproducible start signal • Suitable for use with 5 to 50µL sample loops

Rheodyne 8125 Low-dispersion Microscale Injector Model Mode Features Cat. No. Quantity 8125 Dual C o n t i n u o u s  o w 8125 1 Each

www.thermoscienti c.com/chromatography 4-171 4 -172 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Suitable forpopularRheodyneinjectormodels Injectors for Ports Rheodyne • • • Single-mode sampleinjectordesignedforthecompletefillingmethod Injector Sample 9010 Rheodyne Single-mode sampleinjectordesignedforthecompletefillingmethod Injector Sample 7010 Rheodyne Rheodyne Ports for Rheodyne Injectors Models 7010 and 9010 and 7010 Models Injectors Rheodyne for Ports Rheodyne Injector Sample 9010 Rheodyne Injector Sample 7010 Rheodyne Position sensingswitchprovidesareproduciblestartsignal stator PEEK Compatible withsampleloopsizes5µLto10mL 9010 7010 9010 Needle Port, PEEK Port, Needle 9010 PEEK Port, Filler 9010 Steel Stainless Port, Filler 7010 Model Rheodyne with Use For Model Model Single Single Mode Mode  switch Continuous sensing ow, position Complete lling method Features Features 9010 7010 Cat. No. Cat. No. Cat. 9013 9012 7012 No. Cat. 1 Each 1 Each 1 Each Quantity 1 Each 1 Each Quantity Quantity sample loopsizes5µLto20mL Compatible with: Rheodyne Sample Loops LC Columns and Accessories For Rheodyne sample injectors in stainless steel or biocompatible PEEK

PEEK and Stainless Steel Sample Loops Description Volume ID (mm / in) Cat. No. Quantity Sample loops for 5μL 0.18 / 0.007 7020 1 Each 7010 and 7125 10μL 0.30 / 0.012 7021 1 Each injectors, Stainless Steel 20μL 0.30 / 0.012 7022 1 Each 50μL 0.51 / 0.020 7023 1 Each 100μL 0.51 / 0.020 7024 1 Each 200μL 0.76 / 0.030 7025 1 Each 500μL 0.76 / 0.030 7026 1 Each 1mL 0.76 / 0.030 7027 1 Each 5mL 1.0 / 0.040 7029 1 Each Sample loops for 5μL 0.18 / 0.007 7755-020 1 Each

7725 and 7725i 10μL 0.30 / 0.012 7755-021 1 Each injectors Stainless Steel 20μL 0.30 / 0.012 7755-022 1 Each >>

50μL 0.51 / 0.020 7755-023 1 Each LC Accessories Sample loops for 5μL 0.20 / 0.008 8020 1 Each 8125 injectors, 10μL 0.20 / 0.008 8021 1 Each Stainless Steel 20μL 0.25 / 0.010 8022 1 Each 50μL 0.30 / 0.012 8023 1 Each Sample loops for 2µL Internal 7755-015 1 Each 9010 and 9725 5µL 0.18 / 0.007 9055-020 1 Each injectors, PEEK 10µL 0.25 / 0.010 9055-021 1 Each 20µL 0.25 / 0.010 9055-022 1 Each 50µL 0.51 / 0.020 9055-023 1 Each 100µL 0.51 / 0.020 9055-024 1 Each 200µL 0.51 / 0.020 9055-025 1 Each 500µL 0.76 / 0.030 9055-026 1 Each 1mL 0.76 / 0.030 9055-027 1 Each 5mL 0.76 / 0.030 9055-029 1 Each Sample loops for 5μL 0.18 / 0.007 9055-020 1 Each 9725 and 9725i 10μL 0.25 / 0.010 9055-021 1 Each injectors, PEEK 20μL 0.25 / 0.010 9055-022 1 Each 50μL 0.51 / 0.020 9055-023 1 Each

RheBuild Kits Maintain Rheodyne valves and injectors

RheBuild Kits For Use with Rheodyne Models Cat. No. Quantity 3725/3725i/3725-038/3725i-038 3725-999 1 Each 7010/7000 7010-999 1 Each 7125/7126 7125-999 1 Each 7410 7410-999 1 Each 7520/7526 7520-999 1 Each 7725/7725i/7726 7725-999 1 Each 8125/8126 8125-999 1 Each 9125/9126 9125-999 1 Each

www.thermoscienti c.com/chromatography 4-173 4 -174 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Suitable forpopularRheodyneinjectormodels ReplacementRheodyne Seals Injectors for Rotor For usewithRheodynesampleinjectors Adapter Needle Suction Rheodyne Rheodyne Replacement Rotor Seals for Injectors for Seals Rotor Replacement Rheodyne Needle Suction Adapter Rheodyne 3725/3725i/3725-038/3725i-038 Seals PEEK 9125 9010 8125 7125/7126 7410 7060/7066/9060 7030 7000/7010/7040 Seals Tefzel 8125/8126 7413 7410 7125/7126 7060/7066 7030 7000/7010/7040/7067 Seals Vespel Models Rheodyne with Use For 9725i and 9725 Models Injector Rheodyne with Use For 3725-018 9125-082 9010-051 8125-097 7125-079 7410-075 7060-074 7030-015 7010-071 8125-038 7413-013 7410-038 7125-047 7060-070 7030-003 7010-039 No. Cat. 9125-076 No. Cat. 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity 1 Each Quantity Rheodyne Stators LC Columns and Accessories Suitable for popular Rheodyne injector models

Rheodyne Stators For Use with Rheodyne Models Cat. No. Quantity 7000/7010/7030/7040/7125 7010-040 1 Each 7010-087/7125-081 7010-066 1 Each 7060/7066 7060-039 1 Each 7410/7413 7410-041 1 Each 9010/9030/9125 9125-043 1 Each 9060 9060-016 1 Each 7725 7725-010 1 Each 8125/8126 8125-098 1 Each

Rheodyne Stator Face Assemblies

For Use with Rheodyne Models Cat. No. Quantity >> 3725/3725i/3725-038/3725i-038 3725-039 1 Each LC Accessories 7125 7125-067 1 Each 8125 8125-074 1 Each 9125/9010/9030 8125-094 1 Each 9060 9060-015 1 Each 9725 7725-026 1 Each

Rheodyne Injection Port Needle Cleaner For use with Rheodyne sample injectors

Rheodyne Injection Port Needle Cleaner For Use with Cat. No. Quantity Rheodyne injectors 7125-054 1 Each

Rheodyne Valve Mounting Brackets For use with Rheodyne sample injectors

Rheodyne Injection Port Needle Cleaner For Use with Cat. No. Quantity Angle bracket 7160-010 1 Each Mouting panel 7160 1 Each

www.thermoscienti c.com/chromatography 4-175 4 -176 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Precision machinedfrom316stainlesssteel Fittings Pressure High RheFlex • • • connections metal-free inert, Provide Fittings PEEK Two-Piece RheFlex RheFlex Two-Piece PEEK Fittings PEEK Two-Piece RheFlex RheFlex High Pressure Fittings Pressure High RheFlex Reusable ferruleandnut May beusedon1/16inmetalorplastictubingreliablyupto5000psi Slotted back-sideoftheferruleissqueezeddownontotubeby Replacement ferrules X-long set, Fitting short set, Fitting length standard set, Fitting Type mating conicalsurfaceofthenut 1/16in Ferrule 1/16in Ferrule Type Short Fittings Set Fittings Short Short Fittings Set Fittings Short Extra Long Fittings Set Fittings Long Extra Set Fittings Long Extra Long Fittings Set Fittings Long Set Fittings Long 0.5mm Ferrule for Model 8125 Model for Ferrule 0.5mm 6000-051 6000-066 6000-055 6000-054 No. Cat. Cat. No. Cat. 8125-084 6000-110 6000-210 6000-211 6000-111 6000-262 6000-162 6000-109 6000-209 10 Pack 10 10 Pack 10 10 Pack 10 1 Each 10 Pack 10 5 Pack 1 Each 5 Pack 5 Pack Quantity Quantity 5 Pack 5 Pack 5 Pack 5 Pack Cheminert Model C2 Microbore Injector LC Columns and Accessories Can be used as an injector or switching valve

• 1/16in ttings • 0.010in ports • Available in 6-port or 10-port con gurations • Available with manual or microelectric actuator

Cheminert Model C2 Microbore Injector Description Sample Volume Cat. No. Quantity Model C2 injector, N60 stainless stator, 5μLloop, manual 6 ports C2-1006 1 Each 10 ports C2-1000 1 Each Model C2 injector, N60 stainless stator, 5μL loop, 6 ports C2-1006EH 1 Each microelectric actuator 10 ports C2-1000EP 1 Each

Sample injector loops, stainless steel 2μL CSL2 1 Each

5μL CSL5 1 Each >>

10μL CSL10 1 Each LC Accessories 20μL CSL20 1 Each 50μL CSL50 1 Each 100μL CSL100 1 Each Model C4 injector, PAEK stator, 5μL loop, 6 ports C2-1346EH 1 Each microelectric actuator 10 ports C2-1340EP 1 Each Sample injector loops, PAEK 5μL CZSL5PK 1 Each 10μL CZSL10PK 1 Each 50μL CZSL50PK 1 Each 100μL CZSL100PK 1 Each

Valco Injector Model C6W

Description Volume Cat. No. Quantity Model C6W injector, six 0.016in ports, manual 20μL loop C6W 1 Each Model EPC6W injector, six 0.016in ports, microelectric actuator 20μL loop EPC6W 1 Each Replacement rotor – SSAC6W 1 Each Sample injector loop, stainless steel 2μL SL2CW 1 Each Sample injector loop, stainless steel 5μL SL5CW 1 Each Sample injector loop, stainless steel 10μL SL10CW 1 Each Sample injector loop, stainless steel 20μL SL20CW 1 Each Sample injector loop, stainless steel 50μL SL50CW 1 Each Sample injector loop, stainless steel 100μL SL100CW 1 Each

Valco Accessories

Description Volume Cat. No. Quantity Valco syringe ports 22 ga. needles; 1/16in ttings VISF-1 1 Each Valco syringe ports 22 ga. 2in needles VISF-2 1 Each Valco Nuts and Ferrules 1/16in standard nut ZN1-10 10 Pack Valco Nuts and Ferrules 1/16in long nut LZN1-10 10 Pack Valco Nuts and Ferrules 1/16in SS ferrule ZF1-10 10 Pack

www.thermoscienti c.com/chromatography 4-177 4 -178 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Syringes andneedlesforThermoScientificLCinstruments Syringes HPLC LCQ XSQ LCQ MS LCQ Advantage XP, DECA, LCQ, Instrument Needles Probe ESI Replacement Spectrometry Mass Volume (µL) Scienti Thermo for c Instruments Syringes 1000 500 250 100 25 Volume (µL) Instruments 3000 UltiMate for Syringes 250 500 250 500 2500 Needle Type Needle WPS-3000TXRS the except variants, ACC-3000 and WPS-3000 All WPS-3000TXRS the except variants, WPS-3000 All WPS-3000TXRS the except variants, ACC-3000 and WPS-3000 All variants WPS-3000 All variants WPS-3000 All Instrument Removable Removable – – – Length (mm) Length 50 50 – – – ag Instrument Gauge 22 22 – – – 365RNLT3 365RNLT2 365RNLT1 No. Cat. LCQ LCQ AS1000, AS3000 AS1000, AS3000 AS3000, AS3500 1 Each 1 Each 1 Each Quantity 6822.0005 6822.0004 6822.0003 6822.0002 6822.0001 No. Cat. Cat. No. Cat. 365ILT21 365JLT41 365ILT91 365JLT61 365LLT81 1 Each 1 Each 1 Each 1 Each 1 Each Quantity Quantity 1 Each 1 Each 1 Each 1 Each 1 Each LC Syringes for Manual Injection valves LC Columns and Accessories Easy, accurate and reproducible manual injection

• Square tip to prevent damage to the injector • Wide range of volumes • Precision made from borosilicate glass and stainless steel • Robust design and easy-to-read markings

Gas Tight Syringes for Rheodyne / Valco Injectors Volume (µL) Needle Type Length (mm) Gauge Tip Style Cat. No. Quantity 10 Removable 50 22 90° Blunt End 365DLG21 1 Each 25 Removable 50 22 90° Blunt End 365FLG31 1 Each 50 Removable 50 22 90° Blunt End 365GLG41 1 Each 100 Removable 50 22 90° Blunt End 365HLG51 1 Each

250 Removable 50 22 90° Blunt End 365ILG61 1 Each 500 Removable 50 22 90° Blunt End 365JLG71 1 Each >> LC Accessories Replacement Needles Replacement Needle for Syringe Cat. No. Quantity 365DLG21 365RNL15 5 Pack 365FLG31 365RNL25 5 Pack 365GLG41 365RNL25 5 Pack 365HLG51 365RNL25 5 Pack 365ILG61 365RNL25 5 Pack 365JLG71 365RNL25 5 Pack

Gas Tight Syringes for Rheodyne / Valco Injectors Volume (µL) Needle Type Length (mm) Gauge Tip Style Cat. No. Quantity 10 Fixed 50 22 90° Blunt End 365DL263 1 Each 25 Fixed 50 22 90° Blunt End 365F6315 1 Each 50 Fixed 50 22 90° Blunt End 365G6316 1 Each 100 Fixed 50 22 90° Blunt End 365H6317 1 Each 250 Fixed 50 22 90° Blunt End 365ILG61 1 Each 500 Fixed 50 22 90° Blunt End 365J6319 1 Each

Syringes for Rheodyne / Valco Injectors Volume (µL) Needle Type Length (mm) Gauge Tip Style Cat. No. Quantity 5 Fixed 50 22 90° Blunt End 365CL221 1 Each 10 Fixed 50 22 90° Blunt End 365DL231 1 Each 25 Fixed 50 22 90° Blunt End 365FL241 1 Each 50 Fixed 50 22 90° Blunt End 365GL251 1 Each 100 Fixed 50 22 90° Blunt End 365HL261 1 Each 250 Fixed 50 22 90° Blunt End 365IL271 1 Each 500 Fixed 50 22 90° Blunt End 365JL281 1 Each

www.thermoscienti c.com/chromatography 4-179 4 -180 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • Syringes LC Other Macrovolumesamplingsyringessuitableforliquidorgassamples Luer-Lok syringesidealforpriming Instrumentspeci csyringes Volume (mL) Volume Sampling Volume Macro for Syringes Syringes Priming Luer-Lok for Needles (mL) Volume Syringes Luer-Lok PTFE Volume (µL) Instruments Waters for Syringes 365FL715 365DL991 Replacement Plunger for Syringe Plungers Replacement Volume (µL) Instruments CTC for Syringes 1 1 1 10 2.5 2.5 1 25 5 5 50 10 10 100 25 Needle Type Needle Type Needle Type Needle Type Needle Fxed – Fixed Fixed Fxed – Fixed Fixed Fxed – Fixed Fxed – Fixed – Length (mm) Length Length (mm) Length (mm) Length (mm) Length (mm) Length 50 50 – 70 50 50 – 70 50 50 – 50 50 – 50 – 365RP922 365RP532 No. Cat. Gauge Gauge Gauge Gauge Gauge 22 22s – 26s 22 22s – 26s 22 22s – 22 22s – 22 – 1 Each 1 Each Quantity Tip Style Tip Tip Style Tip Style Tip Style Tip Style Tip 90° Blunt End Blunt 90° Bevel – Bevel – Bevel – End Blunt 90° – Bevel – – Bevel – – – Cat. No. Cat. Cat. No. Cat. No. Cat. No. Cat. No. Cat. 365RNL22 365K3051 365KL531 365BL443 365DL991 365LL375 365LL541 365BL447 365FL715 365M5212 365ML551 365GL810 365N5214 365NL561 365HL331 365PL571 Quantity Quantity Quantity Quantity Quantity 2 Pack 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Detector Lamps LC Columns and Accessories

Detector Lamps for UltiMate 3000 Instruments Description Model Cat. No. Quantity Deuterium Lamp MWD-3000(RS), DAD-3000(RS), VWD-3100 6074.1110 1 Each and VWD-3400RS Tungsten Lamp MWD-3000(RS), DAD-3000(RS), VWD-3100 6074.2000 1 Each and VWD-3400RS

Detector Lamps for Thermo Scienti c Instruments Description Model Cat. No. Quantity Deuterium Lamp SP8400/SP8430/SP8440/SP8450/SP8480/SP8490 DSP-901 1 Each SP8480XR/SP8773XR DSP-907 1 Each Linear UV100/UV200/UV1000/UV2000/UV3000/ DSP-908 1 Each Focus/Spectrochrom

Detector Lamps for Agilent Instruments >>

Description Model Cat. No. Quantity LC Accessories Deuterium Lamp Agilent HP1040/HP1050 (G1306A) DAD/HP 1050 DA DHP-901 1 Each (1050 MWD)/ HP MW (79854A) / HP 1090 (75880A) DAD HP 1080/HP 1081/HP1081B/HP1082B/HP1084/HP1084B DHP-902 1 Each HP 1050 VW (79853C) DHP-903 1 Each HP 8450/8450A DHP-909 1 Each HP 1100 (G1314) VW DHP-910 1 Each HP 1100 (G1315A) DAD DHP-911 1 Each HP 8453 DHP-912 1 Each HP 8452 A DAD/HP 8452A Opt 002 DHP-913 1 Each Xenon Lamp HP 1046/HP1046A DHP-906 1 Each LL Deuterium Agilent 1100 VWD long life DHP-910LL 1 Each Lamp Agilent 1100 DAD long life DHP-911LL 1 Each

Detector Lamps for Merck-Hitachi Instruments Description Model Cat. No. Quantity Deuterium Lamp 101/102/111 DHI-901 1 Each 100-10/100-40/100-50/100-60 DHI-902 1 Each 150-20/200/220/300/330/340/2000/3000/4000/ DHI-903 1 Each L2500/L3000/L4000/L-4500 L4200/L4250/L4500 DHI-908 1 Each LaChrom L4720/L4520/L7400/L450 DHI-910 1 Each Xenon Lamp Hitachi  uorescence detectors F1000/2000/4000 Series DHI-911 1 Each

Our applications team is regularly presenting at key events, download one of our recent posters. www.thermoscienti c.com/chromatography

www.thermoscienti c.com/chromatography 4-181 4 -182 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Detector Lamps for Waters Instruments Waters for Lamps Detector Instruments Varian for Lamps Detector Instruments Shimadzu for Lamps Detector Instruments PerkinElmer for Lamps Detector Lamp Deuterium LL Xenon Lamp Xenon Zinc Lamp Zinc Cadmium Lamp Tungsten Lamp Lamp Deuterium LL Deuterium Lamp Deuterium Lamp Xenon Lamp Tungsten Mercury Lamp Mercury Lamp Deuterium Lamp Deuterium Lamp Deuterium Description Description Description Description Waters Alliance 2487/2488 Alliance Waters 996 Waters 474 only Lamp 470/475/2475 440/441/490 440/441/490 RI/R401/R403/R404 life long Series SPD-10 Shimadzu 2487 Dual Wavelength/2488 Dual 2487 990/991/994 PDA 996 PDA/2996 2486 486 484 480/481/480LC/481LC/Lambda Max/LC1 RF10AX RF10A Shimadzu RF1501.5301/5000 Shimadzu RF540/RF535/RF551/RF500 Shimadzu RF530/RF510 2/2S/10/11Lambda and others 440/441/490 UV/Vis 340/345 ProStar 9050 Star LC5000/LC5500 UV5/2550 UV100/UV200 50/Varichrom UV UV 2050 SPD-M10AVP PDA SPD 10A/SPD 10AS/SPD-10AV/SPD-10AVP SPD 6A/SPD-6AV D300L/UV200S SPD-2A/SPD-3/SPD-4 UV120/UV160/UV160A/UV240/UV260/UV265 DAD 200 Series 2/2S/10/11Lambda and others LC-90/LC-290 Integral 2000/Integral 4000/LC55/LC65/LC85/LC95 360/460/560 3/7/9 Lambda Model Model Model Model DWA-930LL DWA-921LL DWA-929 DWA-923 DWA-913 DWA-912 DWA-911 DSH-918LL DWA-930 DWA-926 DWA-921 DWA-918LC DWA-918 DWA-915 DWA-910 DSH-915 DSH-914 DSH-913 DSH-912 DPE-908 DWA-901 DVA-909 DVA-907 DVA-906 DVA-905 DVA-904 DVA-903 DVA-901 DSH-918 DSH-917 DSH-916 DSH-903 DSH-902 DSH-901 DPE-915 DPE-914 DPE-913 DPE-911 DPE-906 DPE-903 Cat. No. Cat. No. Cat. No. Cat. No. Cat. 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity Quantity Quantity Quantity Pump Spares LC Columns and Accessories

Pump Spares for UltiMate 3000 Instruments Description Model Cat. No. Quantity Maintenance Kits Maintenance Kit for ISO-3100SD ISO-3100SD 6040.1950 1 Each Maintenance Kit for LPG-3400SD LPG-3400SD 6040.1951 1 Each Maintenance Kit for DGP-3600SD DGP-3600SD 6040.1952 1 Each Maintenance Kit for HPG-3x00SD HPG-3x00SD 6040.1953 1 Each Maintenance Kit for LPG-3400RS LPG-3400RS 6040.1954A 1 Each Maintenance Kit for DGP-3600RS DGP-3600RS 6040.1955A 1 Each Maintenance Kit for HPG-3x00RS HPG-3x00RS 6040.1956A 1 Each Maintenance Kit for LPG-3400XRS LPG-3400XRS 6043.1954 1 Each Pistons Pistons, sapphire SD, RS, BM pumps 6040.0042 2 Each Pistons, sapphire LPG-3400XRS pump 6043.0169 1 Each Piston Seals >> Piston Seals, RP SD, RS pumps 6040.0304 2 Each Piston Seals, NP SD, RS pumps 6040.0306 2 Each LC Accessories Piston Seal LPG-3400XRS pump 6043.0295 1 Each Check Valves Check valve, sapphire, cardridge type SD, RS, BX, BM pumps 6041.2300 1 Each Check valve, ZrO ceramics, cardridge type SD, RS, BX, BM pumps 6041.2301 1 Each Check valve, sapphire, cardridge type LPG-3400XRS pump 6043.0145 1 Each

Pump Spares for Thermo Scienti c Instruments Description Model Cat. No. Quantity Pistons Piston Accela pump 00201-11328 2 Each Piston, sapphire Accela pump 00950-01-00126 2 Each Piston Seals Secondary piston seal (wash seal) Accela pump 00950-01-00128 2 Each Piston Seal Black Surveyor LC SFS-220 1 Each Piston Seal Yellow Surveyor LC SFS-220G 1 Each Wash Seal White Surveyor LC SFS-230 1 Each Piston Seal Black Surveyor MS SFS-320 1 Each Piston Seal Clear Surveyor MS SFS-320U 1 Each Wash Seal clear Surveyor MS SFS-330 1 Each Check Valves Outlet Check Valve Accela pump 00950-01-00131 1 Each Inlet Check Valve Accela pump 00950-01-00130 1 Each Inlet Check Valve Assembly – Cartridge Type Surveyor LC SFS-3001 1 Each Outlet Check Valve Assembly – Cartridge Type Surveyor LC SFS-3002 1 Each Inlet/Outlet Check Valve Cartridge Surveyor MS SFS-6001C 1 Each

www.thermoscienti c.com/chromatography 4-183 4 -184 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Pump Spares for Varian Instruments Varian for Spares Pump Instruments PerkinElmer for Spares Pump Instruments Agilent for Spares Pump Outlet Check Valve Assembly Valve Check Outlet Assembly Valve Check Inlet Spares and Valves Check Yellow Seal Piston Black Seal Piston Black Seal Piston Seals Piston Piston Assembly Sapphire Piston Assembly Sapphire Pistons Description Assembly Valve Check Outlet Inlet/Intermediate Check Valve Assembly Spares and Valves Check Yellow Seal Piston LP Black Seal Piston LP Yellow Seal Piston HP Grey Seal Piston HP Seals Piston Sapphire Assembly Piston HP Sapphire Assembly Piston HP Pistons Description Assembly Valve Check Inlet/Outlet Cartridge Valve Check Inlet/Outlet Replacement Spares and Valves Check Black Seal Piston Yellow Seal Piston Seals Piston Piston Assembly Sapphire Piston Assembly Sapphire Pistons Description Series 200, 400, 410, 620, Model 250, Integral 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series 4000 Integral 250, Model 410, 620, 400, 200, Series Model 1090 1090 1100 and 1050 1100 and 1090 1050, 1100 and 1050 1090 Model 2010, 2210, 2510 2210, 2010, 2510 2210, 2010, 2510 2210, 2010, 2510 2210, 2010, 5000, 5500, 5600 2510 2210, 2010, 5000, 5500, 5600 Model SOT-PE3002 SOT-PE3001 SOT-PE320G SOT-PE320 SOT-PE220G SOT-PE220 SOT-PE500 SOT-PE600 No. Cat. SHP-5001 SHP-5002 SHP-420K SHP-220G SHP-400 SHP-200 No. Cat. SVA-3002 SVA-3001 SOT-VA320G SOT-VA320 SOT-VA220 SOT-VA400 SOT-VA200 No. Cat. 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Quantity LC Columns and Accessories Pump Spares continued

Pump Spares for Shimadzu Instruments Description Model Cat. No. Quantity Pistons Piston Assembly Sapphire LC-10 AS, LC-6, LC-6A SOT-SH200 1 Each Piston Assembly Sapphire LC-9, LC-10AD, LC-600 SOT-SH202 1 Each Piston Seals Piston Seal Yellow LC-10 AT SOT-SH-100-01 1 Each Wash Seal White LC-10 AT SOT-SH-100-02 1 Each Piston Seal Grey LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS SOT-SH220 1 Each Wash Seal White LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS SOT-SH220G 1 Each Piston Seal Yellow LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS SOT-SH520G 1 Each Piston Seal Grey LC-9, LC-10AD, LC-600 SOT-SH420 1 Each Piston Seal Black LC-10 ATvp SOT-SH520 1 Each Check Valves and Spares Inlet Check Valve Assembly LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS SOT-SSH3001 1 Each >> Outlet Check Valve Assembly LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS SOT-SSH3002 1 Each Inlet Check Valve Assembly – Cartridge Type LC-9, LC-10AD, LC-600 SSH-6001 1 Each LC Accessories Outlet Check Valve Assembly – Cartridge Type LC-9, LC-10AD, LC-600 SSH-6002 1 Each

Pump Spares for Waters Instruments Description Model Cat. No. Quantity Pistons Piston Assembly Sapphire M510, M590, M600, M610 M6000 SWA-WA200 1 Each Piston Assembly Ruby M510, M590, M600, M610 M6000 SWA-WA200R 1 Each Piston Assembly Sapphire M45, M501 SWA-WA205 1 Each Piston Assembly Sapphire M515 SWA-WA800 1 Each Piston Assembly Sapphire Alliance 2690 SWA-WA900 1 Each Piston Seals Piston Seal Black M45, M501, M510, M590, M600, M610 M6000 SWA-WA220 1 Each Piston Seal Yellow M45, M501, M510, M590, M600, M610 M6000 SWA-WA220G 1 Each Piston Seal Grey M510EF, M590EF, M600EF, M610EF, M6000EF SWA-WA600S 1 Each Piston Seal Black M515 SWA-WA820 1 Each Piston Seal Yellow M515 SWA-WA820G 1 Each Piston Seal Black Alliance 2690 SWA-WA920 1 Each Piston Seal Yellow Alliance 2690 SWA-WA920G 1 Each Check Valves and Spares Inlet Check Valve Assembly M45, M501, M510, M590, M600, M610 M6000 SWA-3201 1 Each Outlet Check Valve Assembly Actuator Type M45, M501, M510, M590, M600, M610 M6000 SWA-3202 1 Each Outlet Check Valve M45, M501, M510, M590, M600, M610 M6000 SWA-3202B 1 Each Inlet Check Valve Repair Kit M510, M590, M600, M610 M6000 SWA-3212 1 Each Outlet Check Valve Assembly Actuator Type M45, M501, M510, M590, M600, M610 M6000 SWA-3402 1 Each Outlet Check Valve Assembly Ball & Seat Type M45, M501, M510, M590, M600, M610 M6000 SWA-3402B 1 Each Inlet Check Valve Assembly M510EF, M590EF, M600EF, M610EF, M6000EF SWA-4107 1 Each Inlet Check Valve Repair Kit M510EF, M590EF, M600EF, M610EF, M6000EF SWA-4123 1 Each Inlet Check Valve Assembly M515 SWA-8001 1 Each Outlet Check Valve Assembly M515 SWA-8002 1 Each Check Valve Cartridge Alliance 2690 SWA-9001 1 Each

www.thermoscienti c.com/chromatography 4-185 4 -186 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories a temperature-controlledconductivitycell. 100 -240VAC, 50–60Hz,autoranging Power Requirements: Weight 10kg(22lbs) 33 ×2440cm(139.515.75in) Dimensions (h×wd): Dionex ICS-900PhysicalSpeci cations • • • • • • Dionex ICS-900KeyFeatures: Thermo Scientific and cationapplications.Thesystemuses designed torunspecificisocraticanion single-channel ionchromatographysystem The DionexICS-900isanintegrated, • • • Routinely AnalyzeMultipleAnionsorCationsin10-15Minutes Systems IonChromatography Dionex ICS-900 damper, electricallyactuatedPEEK reliable dualpistonpump,high-pressurepulse system hasanall-polymericflowpathwitha low noiseandstablebaselines.EachICS-900 Chemical Regeneration(DCR)technologyfor membrane suppressionwithDisplacement USB connectivityforfast,trouble-free Chromeleon ChromatographyDataSystem All-polymeric  owpathwaytoeliminate Dual-piston, serial-pumpingsystemwith Wide pump owraterangetosupport2,3, Dionex MMS300membranesuppression Thermo Scienti c polymeric separation of columns for unparalleled application range broad a with Compatibility Sensitive, stable,heatedconductivitydetectionforpreciseresults instrument connectionandcon guration processing needs and versatilereportstoexceedallyourdata software forfullcontrol,qualityintegration, contamination andcorrosion PEEK and 4mmisocraticanioncationcolumns and ease-of-use with DCRtechnologyfordrift-freebaseline  exibilityandreliability and maximumup-time with anautosampler ™  owpathforlowmaintenancecosts ™ Dionex ™ Dionex ™ MMS ™ Chromeleon ™ ™ 300 valve,and ™ SEallowscontrolofasingleThermoScienti c or loadingconcentratorcolumns. injection parametersforfillingloops provides highflexibilitytoselecttheoptimum chromatograph. Itsnewsoftwarecontrol automated sampleintroductiontothe preparation capabilitiesprovideeasily applications. Therandomaccessandsample designed especiallyforionchromatography free, rugged,automatedsampleloadingdevice The AS-DVautosamplerisalow-cost,metal- 100 -240VAC, 50–60Hz,autoranging Power Requirements: Weight: 16kg(35lbs) 23 ×4556cm(917.522in) Dimensions (h×wd): AS-DV AutosamplerSpeci cations • • • • • • • AS-DV KeyFeatures: Sample Overlapinjectionsforincreased Automatic switchingpowersupplyfor 0.5mL and5.0mLpolymericvialswith Chromeleon controlprovideshigh exibility Random access Optional 6-portor10-portvalveforautomated Full loopandconcentratorloading productivity universal inputvoltage optional ltercaps to selecttheoptimuminjectionparameters sample preparationorinjection

Dionex ICS-900 LC Columns and Accessories Dionex ICS-900 Ion Chromatography System Bundles Description Cat. No. Dionex ICS-900 IC System for Anions with AS-DV Autosampler and Chromeleon Software 078028 Dionex ICS-900 IC System for Cations with AS-DV Autosampler and Chromeleon Software 078029 Contact Customer Service for details of replacement columns and reagents for the Dionex ICS-900

The Anion Bundle includes: The Cation Bundle includes: • Dionex ICS-900 Ion Chromatography System • Dionex ICS-900 Ion Chromatography System • AS-DV Autosampler • AS-DV Autosampler • Chromeleon 7 SE Software • Chromeleon 7 SE Software • AS22 4mm Consumables Bundle: • CS12A 4mm Consumables Bundle: 1- Dionex IonPac AS22 4mm, 064141 1- Dionex IonPac CS12A 4mm, 046073 1- Dionex IonPac AG22 4mm, 064139 1- Dionex IonPac CG12A 4mm, 046074 ™ ™ ™ 1- Thermo Scientific™ Dionex™ AMMS™ 1- Thermo Scientific Dionex CMMS 300 Anion MicroMembrane™ 300 Cation MicroMembrane Suppressor

Suppressor 4mm, 064558 4mm, 064560 >> 1- Dionex AMMS III Regenerant 1- Dionex CMMS III Regenerant LC Accessories Concentrate, 4 pack, 057555 Concentrate, 4 pack, 057556 1- Dionex ICS-900 DCR Anion Regenerant 1- Dionex ICS-900 DCR Cation Regenerant 2L Bottle, 057712 2L Bottle, 057713 1- AS22 Eluent Concentrate, 0670 1- CS12A Eluent Concentrate, 057562

www.thermoscienti c.com/chromatography 4-187 4 -188 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Thermo Scientific short HOTPOCKETmodel.The columns upto150mmcanbeusedwiththe accepts columnlengthsupto300mm,and vertical, orslantposition.Thestandardsize directly ontothecolumn,insitu,horizontal, around sealingmantle.Themantleiswrapped column temperatureusinganovel,soft,wrap- pre-heater. Theinsertsare the actualtemperatureand permanently attachedtothe Temperature ControllerUnit,whichis diameter columns.Thetemperatureissetonthe point aresimultaneously the useofguardcolumnsor your specificcolumn them tobeeasilyremovedorrearrangedfor and ashortversionforcolumnsupto150mm. column combinationsupto300mminlength, available inastandardsizetoaccommodate to temperaturechanges.TheHOTPOCKETis by exploringthesensitivityofaseparation HPLC methodruggednesscanbeinvestigated of HPLCmethodsataccuratetemperatures. 85°C withexcellentcontrol,allowingvalidation temperature rangeofjustaboveambientto The HOTPOCKETColumnHeaterhasa HOT POCKETColumnHeater saving designfortheefficientcontrolofHPLC Temperature Controllershaveaunique,space The ThermoScientific Ef cient, CompactDesign Column HeatingandCoolinginan • • • • • Wrap-around columntemperaturecontrolsystems Temperature Column Controllers and POCKET COOL POCKET HOT inserts areavailablefor controller display. Dualdisplayofbothactualandsetpointtemperature Easytoinstallandusewithavarietyofcolumnlengths rangefromjustaboveambientto85°C HOTPOCKET Explore sampleselectivityandstability rangefrom5°Cto55°C COOL POCKET ™ COOL POCKET configuration. Special ™ HOTPOCKET largerorsmaller displayedontheLED modular, allowing

the userselectedset

the optionaleluent heater/cooler. Both inserts alsoallow ™ Column ™ and onbothsidesofambient

which Columns aresimplyplacedintotheinserts, ispositionedintheopenpartofchannel. around thecolumn or removedthroughthespecialslot of theinsertsmayhavetoberearranged as aguardcolumnoreluentpre-heater, some upon columnlengthandauxiliaryfixturessuch HOT POCKETorCOOLPOCKET. Depending It iseasytoinstallacolumnintothe Rotate theinsertssothatgroovein temperature changesonbothsidesofambient. exploring thesensitivityofyourseparationto and checkHPLCmethodruggednessby methods ataccuratetemperaturesnearambient compounds. ItalsoallowsyoutovalidateHPLC of enantiomersorothercloselyrelated a lowertemperaturemaygivebetterseparation Controller isidealforchiralapplicationswhere of 5°Cto55°C.TheCOOLPOCKETTemperature ambient, withanoperationaltemperaturerange of HPLCcolumnsbothaboveandbelow provides efficientcontrolofthetemperature The COOLPOCKETTemperature Controller COOL POCKETTemperature Controller the channel. are thenrotatedtolockthecolumninto The insulatedmantleiswrapped with aVelcro ™ closure. at oneend. each HOT POCKET Column Heaters, Eluent Preheater/Precooler LC Columns and Accessories and COOL POCKET Chiller Column heating or cooling in a compact, efficient design

• The Eluent Pre-Heater provides a dramatic improvement in chromatography as demonstrated by the USP method for enalapril maleate at 80°C • For preheating or precooling the mobile phase before it enters column • Use in temperatures above 50°C or below 15°C • 0.005in ID

Product Speci cations HOT POCKET COOL POCKET Operating Range 5°C above ambient to 85°C 5°C to 55°C

Display Dual LED displays of actual and set point temperatures in °C

Temperature Accuracy ± 2°C over entire range ± 2°C over entire range >>

Temperature Repeatability ± 1°C ± 1°C LC Equipment Temperature Stability ± 0.1°C ± 0.1°C Time to Stabilization (from ambient) 85°C in less than 30 minutes 55°C in 25 minutes, 5°C in 20 minutes Column Capacity Standard: up to 3/8in OD and up to 300mm in length and end- ttings up to 19mm OD (250mm length column with guard or eluent pre-heater in addition to column) Short (HOT POCKET only): up to 150mm total length (100mm column plus guard or pre-heater) Controller Dimensions 2.8 x 4.0 x 6.5in 2.8 x 4.0 x 6.5in Mantle Dimensions Standard: 1.5 x 1.5 x 17in Standard: 1.5 x 4.0 x 17in Short: 1.5 x 1.5 x 12in Power Cord 3 foot retractable coil cord 3 foot retractable coil cord Weight 1lb enclosure (3lb total with power supply) 2lb enclosure (4lb total with power supply) Power 24 VAC, 25 Watts maximum 15 VDC, 20 Watts maximum

With Eluent Pre-heater Without Eluent Pre-heater

80°C 80°C

oven4 oven3

0 1 2 3 4 5 MIN 0 1 2 3 4 5 MIN

Effect of eluent pre-heater on efficiency Data courtesy of Dr. Richard F. Myer, Quantitative Technologies, Inc., Whitehouse, NJ

HOT POCKET Column Heaters, Eluent Preheater/Precooler and COOL POCKET Chiller Description Cat. No. Quantity HOT POCKET Column Heater 92016 1 Each HOT POCKET Column Heater – short version 92016-150 1 Each COOL POCKET Column Chiller 92017 1 Each Eluent Preheater/Precooler 92018 1 Each

www.thermoscienti c.com/chromatography 4-189 4 -190 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories with lengths upto250mm. with accommodate upto4analyticalcolumns to 150mmwhileColumnOven300can analytical HPLCcolumnswithlengthsup 4 ColumnOven 200canaccommodateupto rates toacceleratethechromatography. particle sizes,allowingtheuseofhigherflow when usingcolumnspackedwithsub2µm to 90°Ccansignificantlyreducebackpressure reproducibility andstability. Temperatures up ColumnOven providesthehighestlevelsof With exacttemperaturecontrolto+/-0.1°C, efficient temperaturecontrolofHPLCcolumns. The ThermoScientificColumnOvendelivers Product Description • • • Exact temperaturecontrolforLCcolumns ColumnOven Thermo Scienti Thermo c ColumnOven Product Speci cations Compatible withCTCautosamplersandThermoScienti cMassSpectrometerionsources Fits uptofourcolumnsinonecompartment Temperature controlfromambientto90°C ColumnOven 300 ColumnOven drivers Software volume) (low Exchanger Heat Exchanger Heat Mount MS ABI Mount MS Thermo Mount CTC 200 ColumnOven Description stability: Temperature Temperature control: supply Power control Remote compartment: x52mm column x62mm 212mm Thermostated shield): heat mounted (with dimensions oven Column Column oven dimensions: Description and driversforThermoScientific to aCTCPAL Optional mountsallowColumnOventobesecured Options standalone version(Windows The optionalsoftwarepackageincludesa Scientific MassSpectrometerionsources. port andcolumn. connection /transferlinebetweeninjection An optionalheatexchangercanbeusedasa and otherinstrumentcontrolsoftware. 66001-030 66001-026 66001-025 66001-024 66001-023 66001-022 66001-021 66001-020 No. Cat. 0.1°C in increments 90°C to Ambient 0.1°C +/- 0.1°C in increments 90°C to Ambient 110 VAC -240 port serial or LAN x21mm x21mm 200mm x 46mm x40mm 212mm 200 ColumnOven ® autosamplerandmanyThermo ™ compatible) ™ XCalibur ™

+/- 0.1°C +/- 110 VAC -240 port serial or LAN x21mm x 21mm 300mm x46mm x40mm 312mm 300 ColumnOven 312mm x 62mm x52mm x62mm 312mm SRS Pro Solvent Recycling System LC Columns and Accessories Reduce mobile phase consumption by up to 90%

• Continuously monitors the output signal of the chromatographic detector, recycling the mobile phase to the solvent reservoir when the baseline is below a certain preset threshold • Easy-to-use software is provided to con gure system parameters, perform on-line monitoring and audit trail facilities • No power adapter is required as the solvent saver is powered directly from the chromatography data system PC through a USB connection • Recycles the mobile phase only if switched on: in case of power failure the valve remains in the waste position and the mobile phase in the reservoir remains uncontaminated • Analog input allows unipolar or bipolar operation of the device within a range of ±1V with (a) - At start of 1st waste run an analog-to-digital converter (b) - After delay • TTL/contact closure for the device can be con gured as start, auto-zero or valve position control input >>

Operational Principle LC Equipment • If the input signal level exceeds this threshold value, the SRS Pro redirects the eluent  ow to waste (a), taking account of the transport time from the detector to the switching valve • When the signal returns below the threshold (b), the SRS Pro again waits for the transport delay and then switches the mobile phase back to the reservoir • Autosampler injection marker connected to the SRS Pro zeroes signal input at the moment of injection

Product Speci cations

Data Rate 1Hz Compatible with: Wetted Material PEEK Any HPLC detector Power Source USB port of PC Max. Pressure 30psi/0.2MPa Requires 1 free USB port, MS-Windows XP/2000/Vista

SRS Pro Solvent Recycling System Description Cat. No. Quantity SRS Pro solvent recycling system 66001-001 1 Each

www.thermoscienti c.com/chromatography 4-191 4 -192 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • • For gas-freeHPLCsolvents Degasser Solvent Pro SDG SDG Pro Used todegasthemobilephaseforHPLCandcanbeemployedinotherapplicationswhere Removes dissolvedgasesfromsolvents Quick equilibrationandshortstartuptimes Reliable, continuousoperation High ef ciencyin-linesystem SDG Pro degasser Pro SDG Description Weight Dimensions Physical Altitude Humidity Relative Ambient Temperature Ambient Conditions Storage Pollution Degree Indoor Use vs. Outdoor Altitude (RH) Humidity Relative Ambient Temperature Ambient Conditions Operating Accuracy Signal Validation Output Installation Over-Voltage Category Sockets Wall Adapter AC supplied using not if Requirement Power Adapter AC supplied using if Requirement Power Power Materials Contacting Solvents Volume Dead Degassing Capacity Maximum Flow Rate Degassing of Mode Channels General Product Speci cations gases mayinterferewiththeuseofsystem(suchasanautotitrator) 2.7kg (6lb). Atypical) (0.5 Amaximum 0.85 at 15 –24 VDC (±3Hz) 60Hz to VAC (±10%), 50 –240 1A, 100 Glass- TePEEK, PTFE, lled AF on channel standard for channel per ~480µL 1mL/min at ~2ppm 10mL/min a tube through uoropolymer permeation Gas 4 independent 250mm (9.8in) 250mm Depth: (2.8in) 73mm Width: (5.0in) 127mm Height: 12000M 0 to condensation) (without RH 80% to 20 +60°C to -20 2 Indoor Meters 2000 0 to condensation) (without %RH 80 to 20 10 35°C to Hg 800mm to 20 from VDC ±0.010 ±1.0% reading of Hg) 800mm at VDC 4.000 Hg; 20mm at (0.100 VDC Hg 800mm to 20 from /1mm absolute Hg 5mVDC II Australia Europe, Continental U.K., America/Japan, North interchangeable: adapter, AC with 4 supplied 66001-010 No. Cat. 1 Each Quantity Thermo Scienti c Liquid Chromatography Reagents LC Columns and Accessories

Introduction to HPLC Ion Pair Reagents

High-purity reagents with the selectivity needed for good separation.

In the past, reverse-phase HPLC analysis of Reverse-phase ion pair highly charged acidic and basic compounds chromatography theories was frustrating and resulted in poor resolution. Two principal theories have been proposed Important biomolecules such as amino acids, to explain reverse-phase ion pair peptides, organic acids, polyamines and chromatography. In the first theory, small catecholamines had to be separated by ion polar ion pair reagents react with the ionized exchange or by suppression techniques. solute, forming neutral ion pairs. The second Thermo Scientific Ion Pair Reagents enable theorizes that an active ion exchange surface you to quickly and efficiently analyze charged is produced in which long chain, nonpolar compounds using reverse-phase techniques. anions and cations are absorbed by the >>

Our ion pair reagents are simply dissolved hydrophobic stationary phase. LC Reagents in the HPLC solvent system, resulting To optimize chromatographic separations in in the formation of stable chromatographic ion pair elution systems, high-purity reagents complexes that can be separated using of exceptional optical transparency are needed. reverse-phase columns. Ion Pair Reagents are specially purified for By using the correct ion pair reagents, ion pair chromatography and provide the you achieve: selectivity needed for good separations. • Increased or decreased retention, permitting controlled selectivity • Resolution of complex ionic mixtures without using ion exchange columns • Improved peak symmetry

www.thermoscienti c.com/chromatography 4-193 4 -194 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 5. The reactionbyproducts(ifany)shouldnot 4. The reactionshouldproceedquicklyand 3. The reactionshouldbereproducibleunder 2. A singlederivativeshouldbeformedper 1. At leastoneacidic,polarfunctionalgroup derivatization protocol: There areseveralrequirementsfor both selectivityandimprovedsensitivity. analytical toolthatcanbeusedtoprovide for aspecificanalyticalprocedure.Itisan compound thathaspropertiesmoresuitable of anexistingcompound,producinganew Derivatization isthechemicalmodification for suitablederivatizationprocedures. degree ofselectivity)establishedtheneed provides highsensitivity(aswellassome The lackofauniversalHPLCdetectorthat Designed toprovideselectivityandimprovesensitivity. HPLC for Reagents and Visualization Derivatization detection ofthesample. interfere withthechromatography, orwith easily undermildconditions. the giventimeandreactionconditions. parent compound. parent compound. must beavailableforreactiononthe artifact formation. are purifiedforchromatography, minimizing techniques. Allcompoundsandformulations reagents forpre-andpost-chromatographic We offeravarietyofHPLCdetection to provideselectivityandimprovesensitivity. post-column) techniquesareusedprimarily acids (Figure2).Post-chromatographic(or and quantificationofopticalisomersamino FDAA (Marfey’s Reagent)allowsseparation and increaseordecreaseretentionofsolutes. improve resolution,peaksymmetry techniques canbeusedtoenhancestability, sensitivity indetection.Pre-column offer morethangreaterselectivityand chromatographic (orpre-columntechniques) been employedwithbothtechniques.Pre- addition, off-lineandon-linereactionshave are bothusedinHPLCderivatization.In Pre- andpost-chromatographictechniques 2.0mL/minute, 25˚C, 340nm 25˚C, 2.0mL/minute, minutes, 45 Bin 40% to 10 gradient: Linear B–acetonitrile. 3.0; pH phosphate, Mtriethylamine A –0.05 Conditions: C18 x4.6mm 100mm on L-DOPA and D- of Separation Derivatization Reagents for HPLC LC Columns and Accessories

Functional Group Description Detection* Page Comments Carboxylic Acid p-Bromophenacylate UV 4-201 Formulation of 1.0mmol/ml p-bromophenacyl O O bromide and 0.005mmol/ml crown ether in acetonitrile; pre-column; nanomole detection R C OH Br C CH2 Br 1-7 levels: λmax = 260nm Primary Amine Dabsyl Chloride Vis 4-204 4-N, N-dimethylaminoazobenzene-4’-sulfonyl p-Bromophenacylate MW 277.94 chloride (dabsyl chloride); pre-column; nanomole N HR 8-14 N N O detection levels: λmax = 436nm H N S O Cl Dabsyl Chloride FDAA, Marfey’s MW Reagent 323.80 UV 4-200 1- uoro-2,4-dinitrophenyl-5-L-alanine amide O O– (FDAA); pre-column; nanomole detection levels: N+ O λmax = 340nm. H 15, 28-29 N For chiral separations of amino acids. NH2

–O N+

O F >> Ninhydrin FDAAO Vis 4-203 Post-column; nanomole detection levels: (Marfey's Reagent) 22

λ = 570nm LC Reagents MW 272.19 OH max Ninhydrin MW 178.14 OH

O

PITC N UV 4-204 Phenylisothiocyanate (PITC); pre-column; C PITC 23-24 S Edman‘s Reagent picomole detection levels: λmax = 254nm MW 135.19

TNBSA EC, UV 4-201 2,4,6-Trinitrobenzene-sulfonic acid (TNBSA); O O OH pre- or post-column; nanomole detection levels – S O O with EC and UV, GC - 0.85V; λ = 250nm25-26 + max N N+ TNBSA O O– MW 293.17

N+ –OO Secondary Amine Ninhydrin Vis 4-203 Post-column; nanomole detection levels: 22 R NH R' (see structure above) λmax = 440nm PITC UV 4-204 Phenylisothiocyanate (PITC); pre-column; 23-24 (see structure above) picomole detection levels: λmax = 254nm *EC = electrochemical; F = fluorescence; UV = ultraviolet; Vis = visible.

References 1. Durst, H.D., et al. (1975). Anal. Chem. 47, 1797. 14. Chang, J.Y., et al. (1981). FEBS Letters 132, 117-120. 24. Scholze, H. (1985). J. Chromatogr. 350, 453-460. 2. Borch, R.F., et al. (1975). Anal. Chem. 47, 2437. 15. Marfey, P. (1984). Carlsberg Res. Comm. 49, 591-596. 25. Caudill, W.L., et al. (1982). J. Chromatogr. 227, 331. 3. Grushka, E. et al. (1975). J. Chromatogr. 112, 673. 16. Jones, B.N. and Gilligan, J.P. (1983). Amer. 26. Caudill, W.L., et al. (1982). Bioanalytical System’s 4. Fitzpatrick, F.A. (1976). Anal. Chem. 48, 499. Biotech. Lab. 12, 46-51. Current Separations 4(4), 59. 5. Nagels, L., et al. (1980). J. Chromatogr. 190, 411. 17. Fiedler, H.P., et al. (1986). J. Chromatogr. 353, 201-206. 27. Andrews, J.L., et al. (1982). Arch. Biochem. 6. Ahmed, M.S., et al. (1980). J. Chromatogr. 192, 387. 18. Fried, V.A., et al. (1985). Anal. Biochem. 146, 271-276. Biophys. 214, 386-396. 7. Pierce Technical Bulletin: Preparation of Phenacyl 19. Jones, B.N., et al. (1983). J. Chromatogr. 266, 471-482. 28. Aberhart, D.J., et al. (1985). Anal. Biochem. 151, and p-Bromophenacyl Derivatives for HPLC. 20. Seaver, S.S., et al. (Sept./Oct. 1984). 88-91. 8. Stocchi, V., et al. (1985). J. Chromatogr. 349, 77-82. Biotechniques 254-260. 29. Szokan, G., et al. (1988). J. Chromatogr. 444, 115-122. 9. Chang, J.Y., et al. (1981). Biochem. J. 199, 547-555. 21. Böhlen, P., et al. (1979). Anal. Biochem. 94, 313-321. 10. Chang, J.Y., et al. (1982). Biochem. J. 203, 803-806. 22. Stein, W.H. and Moore, S. (1949). Cold Spring 11. Vendrell, J., et al. (1986). J. Chromatogr. 358, 401-413. Harbor Symp. Quant. Bio. 14, 179. 12. Chang, J.Y., et al. (1984). J. Chromatogr. 295, 193-200. 23. Heinrickson, R.L. and Meredith, S.C. (1984). Anal. 13. Lin J.K., et al. (1980). Anal. Chem. 52, 630-635. Biochem. 137, 65-74.

www.thermoscienti c.com/chromatography 4-195 4 -196 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories high-purity reagentsforbufferpreparation. consistent analyses,itisessentialtouse acid contamination.Therefore,toachieve acid isasignificantcontributortoamino Unfortunately, forhigh-sensitivitywork,citric alkali salt. prepared eitherwithcitricacidoran Note: Pleaserefertopage3-122fordetailsofreferences gradient elution). introduced totheresin(stepchangesor or lithium)andbythemethodofbufferchanges determined bythecounterionused(sodium physiological fluids.Buffersystemsare acids foundinproteinhydrolyzatesor single- ortwo-columnanalysisofamino The buffersystemsarecompatiblewith displace theaminoacidsintodiscretebands. buffer pHorionicstrength)thatworkto Systems havebeendeveloped(byvarying analytical system,aswelltheinstrumentation. exchange chromatographyhaveinvolvedthe Improvements inaminoacidanalysisbyion Acid inAnalysis Amino Developments for solutionsbelowpH7 used bufferingcomponent,citrate,issuitable 9-15 Themostcommonly .16 Buffersare quantitation aredesirable. and sensitivemethodsforseparation protein chemistrytoolsavailable,morerapid amino acidanalysisisoneofthebasic for theseparationofaminoacids.Because Alternatives toionexchangeareavailable o-phthalaldehyde (OPA) eliminate in-linederivatization. These derivativesarestableenoughto phenylthiocarbonyl derivativesviaHPLC. separation andquantitationoftheresulting with phenylisothiocyanate(PITC)andthe quantitative derivatizationofaminoacids techniques. Othermethodsincludethe analysis timethanpost-columnderivatization methods offergreatersensitivityandshorter amino acidspriortoHPLCanalysis.Both involve theformationofdansyl Two ofthemostwidelyusedthesemethods reverse-phase HPLChavebeendeveloped. column derivatizationmethodsusing 20-24 17 derivativesof Severalpre- 18-19 or Sample Preparation and Hydrolysis LC Columns and Accessories

The extraction and purification of proteins play Hydrolysis and chlorotyrosine.27 This procedure minimizes an important role in determining amino acid Most protein samples require some form of decomposition of reduced S-carboxymethylcysteine content. These methods are based on one or chemical treatment before their component and analyzes S-carboxymethylated proteins. more of their physical characteristics (e.g., amino acids are suitable for analysis. Protein Hydrolysis is conducted at 110°C (with the solubility, molecular size, charge, polarity and and peptide samples must be hydrolyzed to temperature accurately controlled) for 20-70 specific covalent or noncovalent interactions). free amino acids from peptide linkages. Acids hours by Moore and Stein’s method.28 After hydrolysis, residual HCI is removed in a rotary The techniques commonly used to (usually HCI) are the most widely used agents evaporator. The residue is dissolved in water separate proteins and peptides include: for hydrolyzing proteins. A simplified hydrolysis procedure involves and brought to the appropriate pH for addition • Reverse-phase HPLC 28 refluxing the protein with excess HCI, then to the analyzer column. • Polyacrylamide gel electrophoresis removing the excess acid in vacuum.26 The • Gel ltration lyophilized protein then is suspended in • Ion exchange chromatography constant boiling 6 N HCI and introduced into • Af nity chromatography the hydrolysis tube. The sample is frozen by

• The table below provides a more detailed list immersing the tube in dry ice and acetone. >> of methods for fractionating peptide mixtures.25 Before sealing, the tube is evacuated to avoid formation of cysteic acid, methionine sulfoxide LC Reagents

Methods for the fractionation of peptide mixtures. Technique Properties of Peptide Molecules Exploited Centrifugation Solubility Size exclusion chromatography Size Ion exchange chromatography Charge, with some in uence of polarity Paper electrophoresis Charge and size Paper chromatography Polarity Thin layer electrophoresis Charge and size Thin layer chromatography Polarity Polyacrylamide gel electrophoresis Charge and size High-performance liquid chromatography (HPLC) Polarity Gas chromatography Volatility of derivatives Counter-current extraction Polarity; sometimes speci c interactions Af nity chromatography Speci c interactions Covalent chromatography or irreversible binding Disul de bond formation; reactivity of homoserine lactone

References 1. Braconnot, H. (1820). Ann. Chem. Phys. 13, 113. 12. Thomson, A.R. and Miles, B.J. (1964). Nature 203, 483. 22. Turnell, D.C. and Cooper, J.D.H. (1982). CIin. Chem. 28, 2. Mulder, G.J. (1848). J. Prakt. Chem. 38, 294. 13. Robinson, G.W. (1975). J. Chromatogr. 3, 416. 527-531. 3. Kjeldahl, J.G.C.T. (1883). Z. Anal. Chem. 22, 366. 14. Benson, J.V. (1972). Anal. Biochem. 50, 477. 23. Umagat, H., et al. (1982). J. Chromatogr. 241, 324-334. 4. Rattenbury, J.M. (1981). Amino Acid Analysis, Ellow 15. Benson, J.V. and Patterson, J.A. (1965). Anal. Chem. 24. Jones, B.N. and Gilligan, J.P. (1983). J. Chromatogr. 266, Horwood, publisher, Chicester, England, Chapter 1. 37, 1108. 471-482. 5. Martin, A.J.P. and Synge, R.L.M. (1941). Biochem. J. 16. Bates, R.G. and Pinching, G.D. (1949). J. Amer. 25. AlIen, G. (1981). Laboratory Techniques in 35, 1358. Chem. Soc. 71, 1274. Biochemistry and Molecular Biology, T.S. Work 6. Elsden, S.R. and Synge, R.L.M. (1944). Proc. 17. Jones, B.N. and Gilligan, J.P. (1983). American and R.H. Burdon, Eds., Elsevier/North-Holland, Biochem. Soc. IX. Biotech. Lab. December, 46-51. Biomedical Press. 7. Synge, R.L.M. (1944). Biochem. J. 38, 285. 18. Weidmeier, V.T., et al. (1982). J. Chromatogr. 231, 410-417 26. Moore, S. and Stein, W.H. (1963). Meth. Enzymol. 6, 819. 8. Stein, W.H. and Moore, S. (1949). Cold Spring Harbor 19. DeJong, C., et al. (1982). J. Chromatogr. 241, 345-359. 27. Eveleigh, J.W. and Winter, G.D. (1970). Protein Symposia Quant. Biol. 14, 179. 20. Lindroth, P. and Mopper, K. (1979). Anal. Chem. 51, Sequence Determination, S.B. Needleman, Ed., 9. Liao, T.H., et al. (1973). Anal. Chem. 45, 2286. 1667-1674. Springer-Verlag, pp. 92-95. 10. Hare, P.E. (1972). Space Life Sciences 3, 354. 21. Jones, B.N., et al. (1981). J. Liq. Chromatogr. 4, 565-586. 28. Moore and Stein, op. cit. 11. Dus, K., et al. (1966). Anal. Biochem. 14, 41.

www.thermoscienti c.com/chromatography 4-197 4 -198 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories X intheorderingtableindicatesthathazardousshippingcharges apply. • • • • Properties ofTriethylamine • • • in improvedpeakresolution. HPLC separations.Bypairingwithpeptides,iteffectivelysharpenspeaks,resulting Triethylamine isanion-pairingreagentthataltersselectivityinreverse-phase Ideal forHPLCseparationandanalysisofpeptides (TEA) Triethylamine X intheorderingtableindicatesthathazardousshippingchargesapply. 3. Bennett,H.P., etal.(1981).Biochemistry. 20, 2. Bennett,H.P.J., etal.(1980).J.LiquidChromatogr. 3, 1. Hearn,M.T.W. andHancock,W.S. (1979).Trends4, Biochem.Sci. • • • • • • Ion-pair reagentforthereverse-phaseHPLCseparationofproteins andpeptides Acid Heptauorobutyric Ion PairHPLC Reagents Triethylamine (TEA) Triethylamine Hepta Acid uorobutyric Density 0.726g/mL Density Molecular weight101.19 Molecular formulaC Alternate namesTEA,Diethylethanamine Has alowUVabsorbancetoprovidethemostsensitivedetectionacross Packaged inamberglassbottleswithprotectivePTFE-lined uorocarboncaps 99.5% triethylaminepurity, allowingsensitivepeptidedetectionatlowUV Clear, colorlessliquid Packaged undernitrogeninamberglassampulesorbottles B.P.120°C 1.645 Density: Sequencing reagentforclassicalandautomatedEdmandegradationofpeptides andproteins Typical purityis99.7%byGC;<0.1%water Triethylamine, Sequencing Grade Hepta Acid, Grade HPLC uorobutyric Triethylamine, HPLC Grade HPLC Triethylamine, Hepta Acid, Sequencing Grade uorobutyric Description Description all wavelengths for reagentintegrity wavelengths inreverse-phaseHPLCpeptideseparationsystems 6 H 15 N 4530-4538. 2(a);0.000MTEA2(b);0.010M2(c);0. 0 Min3 1353-1366. 100g 10 x1mL ampules 25g 100mL Quantity Quantity

2,4-dimethylaniline N58-N62. X X 0 Min3 TS-25108 TS-53104 TS-53101 TS-25003 Cat. No. Cat. No. Cat.

2,4-dimethylaniline 1 Each 1 Pack 1 Each 1 Each Quantity Quantity 2(a);0.000MTEA2(b);0.010M2(c);0.020M2(d);0.030M 0 Min30 0 Min3 020M TEA2(d);0.030M

2,4-dimethylaniline 2,4-dimethylaniline 0 Min3 Min3

2,4-dimethylaniline 2,4-dimethylaniline 0 Min30

2,4-dimethylaniline Min3

2,4-dimethylaniline Formic Acid Ampules LC Columns and Accessories Well-suited for HPLC and mass spectrometry applications

Formic acid is a component found in reverse-phase mobile phases to provide protons for LC/MS analysis. The presence of a low concentration of formic acid in the mobile phase is also know to improve the peak shapes of the resulting separation. Unlike tri uoroacetic acid (TFA), formic acid is not an ion-pairing reagent, and it does not suppress MS ionization of polypeptides when used as a mobile phase component. • Prescored, nitrogen- ushed, amber glass to protect formic acid from light and moisture • 99% purity for consistent LC baselines and no interference introduced into LC and mass spectrometry applications • Convenient format simpli es preparation of gradient and isocratic mobile phases containing 0.1% (v/v) formic acid in water or acetonitrile • Contents of a single vial in a nal volume of 1L solvent yields a mobile phase of the most

common formic acid concentration >> LC Reagents Formic Acid Ampules Description Quantity Cat. No. Quantity Formic Acid 99+% 10 x 1mL ampules TS-28905 1 Each

For complex peptide separations, the key to success can be to vary selectivity. Varying mobile phase composition on the same column can change selectivity enough to resolve peptide that would otherwise overlap. The TFA concentration is usually specified as 0.1% for reverse-phase HPLC of peptides. For reproducible separations from run-to-run or from lab-to-lab, it is essential to make concentrations the same.

www.thermoscienti c.com/chromatography 4-199 4 -200 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • chromatography quickandeasy Makes separationandquantitationofopticalisomersaminoacids byreverse-phase Reagent Marfey’s FDAA, X intheorderingtableindicatesthathazardousshippingchargesapply. • • • Routinely usedion-pairingagentinreversed-phasepeptideseparations Triuoracetic Acid (TFA) HPLC for Reagents and Visualization Derivation FDAA, Marfey’s Reagent Marfey’s FDAA, Tri (TFA) Acid uoracetic Derivatives canbedetectedbyUVat340nmwithpicomolesensitivity Derivatives haveanabsorptioncoef cientof~3x10 Optical isomersofaminoacidsderivatizationcompleteinjust90minutes Choice offormatsforconvenience:1mLampulescanprepare1L0.1%v/vTFA solutionfor High-performance packaging:PackagedundernitrogeninamberglasswithprotectiveTFE-lined Purity: >99.5%TFA andexceptionalclarityallowssensitive,nondestructivepeptidedetection Tri Acid,uoracetic Sequencing Grade Tri Acid,uoracetic Sequencing Grade Tri Acid,uoracetic Sequencing Grade FDAA, Marfey’s Reagent FDAA, Tri Acid,uoracetic Sequencing Grade Description Description Absorbance @ 340 nm the mobilephaseinreverse-phasechromatographymoments  uorocarboncapstoensureTFA integrity at lowUVwavelengths 0 0 3 40 30 20 10

L-Aspartic Acid L-Glutamic Acid D-Aspartic Acid D-Glutamic Acid L-Alanine Excess Reagent D-Alanine L-Methionine

D-Methionine L-Phenylalanine 1g 10 ×1mL 100g 50mg 500mL Quantity Quantity D-Phenyalanine 4

X X X X TS-28902 TS-28904 TS-28903 TS-48895 TS-28901 Cat. No. Cat. No. Cat. 1 Each 1 Pack 1 Each 1 Each 1 Each Quantity Quantity p-Bromophenacylate Reagent LC Columns and Accessories Gives quantitative yields with few or no side reactions

• Premixing of phenacylbromide and crown ether is not necessary • Derivatization is both rapid and quantitative, with yields of >95% in 15 to 20 minutes at 80°C • Excess reactants do not interfere • Large excess of alkylating reagent is not necessary • Small amounts of water or alcohol do not interfere • If isolation is desired, products are usually crystalline

p-Bromophenacylate Reagent Description Quantity Cat. No. Quantity p-Bromophenacylate Reagent 10mL TS-48891 1 Each 1. Durst, H.D., et al. (1975). Anal. Chem. 47, 1797.

2. Borch, R.F., et al. 1975). Anal. Chem. 47, 2437. 3. Grushka, E., et al. (1975). J. Chromatogr. 112, 673. >> 4. Fitzpatrick, F.A., et al. (1976). Anal. Chem. 48, 499. LC Reagents

TNBSA (Trinitrobenzene Sulfonic Acid) An excellent choice for spectrophotometric detection

• Couples with primary amines, sulfhydryls and hydrazides in aqueous solution at pH 8, without undesirable side reactions • Excellent for solution or solid phase analysis • Suitable for qualitative and quantitative estimation of biomolecules; including amino acids, eptides or proteins • Chromogenic, Omax = 335nm • Reacts readily with primary amino groups of amino acids in aqueous format at pH 8 to form yellow adducts • No colored derivatives are formed with secondary amino acids proline and hydroxyproline • Colored derivatives are monitored at 345nm and have extinction coef cients in range of 1-1.5 x 104

TNBSA Description Quantity Cat. No. Quantity TNBSA 100mL X TS-28997 1 Each 1. Goodwin, J.F. and Choi, S-Y. (1970). Clinical Chemistry. 16, 24-31. 2. Snyder, S.L. and Sobocinski, P.Z. (1975). Anal. Biochem. 64, 284-288. 3. Drozdovskaya, N.R., et. al. (1982). FEBS Lett. 150, 385. 4. Takahashi, S., et al. (1984). Chem. Lett. (Jpn). 1, 127 X in the ordering table indicates that hazardous shipping charges apply.

www.thermoscienti c.com/chromatography 4-201 4 -202 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories maximum storagelifeofsixmonths.Storefrozenbetweenuses. When keptfrozen,anunopenedvialhasindefinitestoragelife.Oncetheseal isbroken,thereagenthasa L-Phenylalanine, L-Proline,L-Serine,L-Threonine,L-Tyrosine, L-Valine. Glycine, L-Histidine,L-Isoleucine,L-Leucine, L-alanine, Ammonia[(NH4)2SO4],L-Arginine,L-AsparticAcid,L-Cystine,L-Glutamic The followingaminoacidsareincluded inAminoAcidStandardH: • • • High-purity calibrationstandardforproteinhydrolysates Acid H Standard Amino 4. Hurley, J.B.,etal.(1984).IsolationandcharacterizationofacDNAcloneforthesubunitbovine retinaltransducin.Proc.Natl. 3. Blankenship, etal.(1989).High-sensitivityaminoacidanalysisbyderivatizationwitho-Phthaldialdehyde and9-Fluorescence 2. Eveleigh, J.W. andWinter,1. G.D.(1970).ProteinSequenceDetermination,EdNeedleham,S.B.,Springer-Verlag, pp.92-95. • • • Sequencing-grade reagentfortotalproteinhydrolysis Acid Hydrochloric (6N) Boiling Constant Reagents Hydrolysis Amino Acid Standard H Standard Acid Amino Acid Hydrochloric (6N) Boiling Constant With theexceptionofcystine,eachaminoacidissuppliedataconcentration Molar concentrationveri edbyconventionalaminoacidanalysismethods Uses L-formcon gurationtopermitstandardizationofmicrobialandotherassays Packaged inprescoredampulestoeliminatecontaminationandensureproductintegrity Specially puri edtogiveninhydrin-negativeblankonhydrolysis Hydrolyzes peptidesin6hoursat150°C Amino Acid Standard H Standard Acid Amino 6N Acid Hydrochloric Description Description of 2.5µmoles/mLin0.1NHCl mercaptoethanol reagent.J.Biol.Chem.July25,6248-6251. Lee, K.etal.(1979).Derivatizationofcysteineandcystineforfluorescenceaminoacidanalysiswith theo-Phthaldialdehyde/2- Acad. Sci.USA.81,6948-6952. detection: applicationsinproteinstructuredetermination.Anal.Biochem. 10 ×1mL 10 ×1mL Quantity Quantity L-Lysine•HCl, L-Methionine, 178, 227-232. TS-20088 TS-24308 Cat. No. Cat. No. Cat. 1 Pack 1 Pack Quantity Quantity Amino Acid Detection Reagents LC Columns and Accessories Ninhydrin The reagent of choice for detection of amino acids

• Used in amino acid chromatography • Offers superb color response and low blank • Inde nitely stable and requires no refrigeration

Ninhydrin Description Quantity Cat. No. Quantity Ninhydrin 500g TS-21003 1 Each 1. Stein, W.H. and Moore, S. (1949). Cold Spring Harbor Symp. Quant. Biol. 14, 179.

2. Moore, S. (1968). Amino acid analysis: aqueous dimethyl sulfoxide as solvent for the ninhydrin reaction. J. Biol. Chem. 243(23), 6281-6283.

3. James, L.B. (1978). Amino acid analysis: ninhydrin reaction with titanous chloride. J. Chromatogr. 152, 298-300. >> Indefinitely stable. No refrigeration required. Keep bottle tightly sealed. Avoid exposure to direct sunlight and ammonia. LC Reagents

www.thermoscienti c.com/chromatography 4-203 4 -204 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Evert,R.F. (1986).Anal.Biochem.154,431-435. 3. Janssen,etal.(1986).Chromatogr. 22(7-12). 2. Scholze,H.(1985).J.Chromatogr. 350,453-460. 1. Heinrikson,R.L.andMeridith,S.C.(1984).Anal.Biochem.136,65-74. • • • by reverse-phaseHPLC High-purity reagentforpre-columnquantitativederivatizationof amino acids (Phenylisothiocyanate) PITC X intheorderingtableindicatesthathazardousshippingchargesapply. 8. Stocchi,V., etal.(1985).J.Chromatogr.349, 77-82. 7. Chang,J.Y., etal.(1983).Methods.Enzymol.92,41-48. 6. Lin,J.K.,etal.(1980).Clin.Chem.26,579-583. 5. Vendrell, J.etal.(1986).Chromatogr. 358,401-413. 4. Chang,J.Y., etal.(1981).FEBSLett.132,117-120. 3. Chang,J.Y. (1984).J.Chromatogr. 295,193-200. 2. Chang,J.Y., etal.(1982).Biochem.J.199,803-806. 1. Chang,J.Y., etal.(1981).Biochem.J.199,547-555. • • • • • Recrystallized twice Chloride Dabsyl Reagents Derivatization Pre-Column High-Purity Resulting phenylthiocarbamylderivativescanbeseparatedandquanti edin30minutes Reacts readilywithaminoacidsin5to10minutesatroomtemperature Also knownasEdman’s Reagent Analysis ofaminoacidneurotransmittersinmousebrain Analysis ofphosphoaminoandaminoacidamides Analysis ofpeptidesanddeterminationC-terminalsequencepolypeptides Analysis of10-30ngproteinhydrolysates For theprecolumnderivatizationanddetectionofaminoacidsinvisiblelightdown sulfonyl chloride) Dimethylaminoazobenzene-4- (4-N,N Chloride Dabsyl PITC (Edman’sPITC Reagent) Description (Phenylisothiocyanate) PITC Description Chloride Dabsyl using reverse-phaseHPLCtoproducestableproductswithallaminoacidsincludingproline to sub-picomolarlevels 500g Quantity 10 ×1mL Quantity X TS-21720 Cat. No. Cat. TS-26922 No. Cat. 1 Each Quantity 1 Pack Quantity HPLC and Spectrophotometric Grade Solvents LC Columns and Accessories Ultrapure solvents are carefully packed for thorough protection

• Distilled in glass, ltered through 0.2µm TFE membranes and packed in solvent-rinsed, amber glass bottles • TFE-lined screw caps seal bottles

Acetonitrile, HPLC Grade, Physical Dimethylsulfoxide (DMSO), Sequencing Properties Grade, Physical Properties

• UV Cutoff: 190nm • C2H6OS • Optical Absorbance: <0.02 at 220nm • Purity (GC): >99.5% • Refractive Index at 25°C: 1.342 • MW: 78.13 Water, HPLC Grade, Physical Properties • Density: 1.101 • • UV Cutoff: 190nm Water: ≤0.2%

• Optical Absorbance: <0.005 at 220nm Pyridine >> LC Reagents • Refractive Index at 25°C: 1.332 • C5H5N Dimethylformamide (DMF), Sequencing • Purity (GC): ≥99% Grade, Physical Properties • MW: 79.10

• HCON(CH3)2 • Density: 0.978 • Purity (GC): ≥99% • B.P. 115°C • MW: 73.09 • Density: 0.944 • B.P. 153°C • Water: 0.1%

HPLC and Spectrophotometric Grade Solvents Description Quantity Cat. No. Quantity Acetonitrile 1L X TS-51101 1 Each Water 1L TS-51140 1 Each Dimethylformamide (DMF) 50mL X TS-20673 1 Each Dimethyl sulfoxide (DMSO) 950mL X TS-20688 1 Each Pyridine 100g X TS-25104 1 Each X in the ordering table indicates that hazardous shipping charges apply.

www.thermoscienti c.com/chromatography 4-205 4 -206 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • • • • • Heavy peptidemixtureforcolumnassessmentandpredictionof retentiontimes Mixture Calibration Time Retention Peptide 5. Guo,D.,etal.(1987).J.Chromatogr. 386,205-222. 4. Mant,C.T. andHodges,R.S.(1986).L.C.MagazineLiq.Chrom.HPLC4(3),250. 3. Guo,D.,etal.(1986).J.Chromatogr. 359,519-532. 2. Guo,D.,etal.(1986).J.Chromatogr. 359,499-517. 1. Guo,D.,etal.(1985).ProceedingsoftheNinthAmericanPeptideSymposium,PublishedbyThermoFisherScientific, variation, columnaging,n-alkylchainlengthvariationandliganddensity. decreases beyond20residues.To ensureaccuracy, apeptide standard is used to correct for instrument sizethe with increases of the molecule. The accuracy of predictingsignificantly time retention peptide weight ofthepeptide.Theeffectonretentionisrelativelyunimportantwithasmall peptide,butit amino acidresidueandthepeptideterminalgroups.Retentiontimeisdependent uponthemolecular Retention timesarepredictedbytotalingthevalues that represent the contribution in minutes ofeach • • • • • • • • composition upto20residuesinlength Allows accuratepredictionofelutiontimeforpeptidesknown amino acid Standard Retention Peptide Standards Peptide Evaluate reverse-phasesupportsofvaryingn-alkylchainlengthsandliganddensities Compare reverse-phasecolumnsfromdifferentmanufacturers Monitor columnperformance:ef ciency, selectivityandresolutionduringcolumnageing Predict theHPLCretentiontimeforpeptidesofknownaminoacidcompositiononreverse Determine therelativeorderofpeptideelutionacomplexmixture Increase theef ciencyofpredicting peptideelutionpro les Simplify identi cationofspeci cpeptidesinacomplexmixture Save timeinpeptidepuri cation Internal standardtonormalisefor variationinretentiontimesandpeakintensitiesbetweenruns Optimization ofscheduledMSacquisition windowsforimprovingquanti cationandincreasedmultiplexing Prediction ofpeptideretentiontime fromsequenceusingcalculatedhydrophobicityfactor Prediction ofpeptideretentionacross multipleinstrumentplatforms Assessment ofchromatographyand MSinstrumentperformance Peptide Retention Standard, S1-S5 Standard, Retention Peptide Rockford, Illinois,page23. Mixture, 5pmol/µL Mixture, Calibration Time Retention Peptide 0.5pmol/µL Mixture, Calibration Time Retention Peptide Description Mixture Calibration Time Scienti Retention Thermo c Peptide Description Standard Retention Peptide phase HPLCcolumns 1 vial 1 Quantity 200µL 50µL Quantity TS-31700 Cat. No. Cat. TS-88321 TS-88320 No. Cat. 1 Each Quantity 1 Each 1 Each Quantity Dionex Ion Standard Concentrates LC Columns and Accessories • The Dionex IonPac ready-to-use ion standards are designed for routine anion or cation determinations • All standards are traceable to NIST Standard Reference Materials • Shipped with a Certi cate of Analysis verifying the concentration

Description Volume (mL) Cat. No. Combined Five Anion Standard 100 037157 Combined Seven Anion Standard I 50 056933 Combined Seven Anion Standard II 100 057590 Fluoride Standard (1000mg/L) 100 037158 Chloride Standard (1000mg/L) 100 037159 Sulfate Standard (1000mg/L) 100 037160 Combined Six Cation Standard-I 50 040187 Combined Six Cation Standard-II 50 046070

>> LC Reagents MS Standard Concentrates • MS standards for validation and calibration of the MSQ System

Description Cat. No. Kit of 2 standards for performance validation in ESI and APCI modes 061496 Standard for Mass Calibration (All MSQ Models) 062917

Perchlorate Internal Standard • For the quanti cation of perchlorate at low parts-per-trillion levels using mass spectrometric detection. Contains stable-labeled perchlorate (1mg/L).

Description Cat. No. Perchlorate-ISTD. 18O Internal Standard for Perchlorate 062923

www.thermoscienti c.com/chromatography 4-207 4 -208 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories • Reagents Pairing Ion • • Haloacetic Acid Standards Internal Highly puri edion-pairingreagentsareusedinmobilephaseionchromatography(MPIC), Stable-labeled internalstandardspreparedinMTBE(methyl-t-butylether),1mLampuleplus Ready-to-use internalstandardsforhaloaceticacidanalysisusingelectrospray-mass Octanesulfonic acid (OSA) acid Octanesulfonic (HSA) acid Hexanesulfonic (TPAOH) hydroxide Tetrapropylammonium (TBAOH) hydroxide Tetrabutylammonium Description 1 1 MtBE in 1000mg/L Standard, -2-13C Acid Internal Trichloroacetic 1 MtBE in 1000mg/L Standard, -2-13C Acid Internal Dichloroacetic 1 MtBE in 1000mg/L -1-13C Standard, Acid Internal Monobromoacetic MtBE in 1000mg/L Standard, -2-13C Acid Internal Monochloroacetic Description combining reversed-phaseion-pairchromatographywithchemicalsuppression empty vial spectrometric detection 500 500 500 500 (mL) Volume (mL) Volume 035362 035361 035363 035360 No. Cat. No. Cat. 069409 069408 069407 069406 LC and LC-MS Instrument Accessories, LC Columns and Accessories Consumables and Spare Parts

Our liquid chromatographs and mass spectrometers advance scientific knowledge, enable drug discovery, improve manufacturing processes, and protect people and the environment. When your instrument is maintained with Thermo Scientific parts, you can expect the best results and highest productivity, keeping your research or processes moving smoothly.

>> LC and LC-MS Instrument Parts

www.thermoscienti c.com/chromatography 4-209 4 -210 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories keeping fullycompatibletoconventionalLC. Systems offersindustryleadingversatilitycoveringthemaximumrange ofUHPLC applications,while all chromatographicperformancechallenges.Withanoutstandingproduct portfolio, UltiMate3000RS Precision-engineered instrumentation,advanceddataprocessing,andhighly optimized chemistriesmeet UltiMate 3000RapidSeparation(RS)Systemsprovidesunrivaledperformance and flexibilityinUHPLC. Systems Rapid LC (RS) Separation applications andprovideUHPLCcompatibility–allowingyoutomovewhenever youareready. gradient responsetimes,aswellsuperiormixingperformance.Theyofferfullsupport ofallHPLC components providelowextra-columnandgradientdelayvolumeforhighseparation efficiencyandlow UltiMate 3000Standard(SD)Systemsaredesignedtomeetcurrentandfuturechallenges. System (SD) Systems Standard LC The ThermoScientific Basic Automated System Portfolio Scienti Systems Thermo 3000 c Dionex UltiMate a ruggedsampleinjectionprinciplewithpowerfulcolumnoven. Autosampler ColumnCompartmentisattheheartofsystem.Itsuniqueinstrument designcombines and ease-of-usewithroutineLCapplications,alsooffersfullUHPLCcompatibility. TheACC-3000 ™ Dionex ™ UltiMate ™ 3000BasicAutomatedSystemisoptimizedforreliability Biocompatible Rapid Separation (BioRS) LC Systems LC Columns and Accessories UltiMate 3000 Biocompatible Rapid Separation (BioRS) Systems are powered by UltiMate 3000 Rapid Separation technology to support the high pressures required for the separation of bioanalytes on high resolution bio UHPLC columns. This state-of-the-art technology combined with a biocompatible, low-dispersion flow path, provides the highest peak capacity and sensitivity for complex samples, whether proteins, peptides, or biotherapeutics. Bio UHPLC enables faster gradients and shorter run times compared to conventional Bio LC analysis. This results in increased sample throughput in typical modes of biochromatography.

x2 Dual Rapid Separation LC Systems >>

UltiMate 3000 x2 Dual LC Systems are the ideal choice to maximize sample throughput of conventional LC and LC-MS Instrument Parts HPLC and UHPLC separations by combining the flexibility of ternary solvent selection with the productivity and performance of x2 Dual technology. Advanced chromatographic techniques such as parallel, tandem, and two-dimensional LC become a realistic and practical option, even at ultrahigh speed. Operate conventional or ultrahigh speed applications, increase sample throughput, achieve higher chromatographic resolutions, or automate tedious sample preparation steps such as analyte enrichment or matrix elimination.

XRS LC Systems UltiMate 3000 XRS Systems are highly LC/MS optimized quaternary UHPLC systems and the best choice for ultrahigh resolution and high speed UHPLC MS-frontend solutions. This system is the ideal solution for high sample throughput and ultra high resolution chromatography.

www.thermoscienti c.com/chromatography 4-211 4 -212 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories UltiMate 3000 Solvent Racks Solvent 3000 UltiMate Instruments 3000 UltiMate Accessories, Consumables for and Spare Parts HPLC Troubleshooting Guide poster Guide Troubleshooting HPLC toothing torque with capillaries Viper for tool Installation Menu pen systems 3000 UltiMate for kit Drainage Description Instruments 3000 UltiMate with Use the for Accessories External power supply unit supply power External SRD for Kit Upgrade Channel Degasser guides) 10 retaining and caps (5 bottles solvent for guides retaining and Caps reservoirs solvent for cap Bottle head pump to degasser analytical from line supply Solvent (analytical) line supply Solvent Description Racks Solvent 3000 UltiMate for Parts Spare and Consumables for iPhone, Android and Windows phones. for iPhone,Android andWindows Also useourHPLCTroubleshooting GuideasaMobileApp 1 1 1 1 Quantity 1 1 1 4 1 1 Quantity 6040.0050 6040.2314 6300.0100 6040.0005 No. Cat. 6510.0004 6035.0089 6030.9101 6270.0013 6030.2546 6030.2548 No. Cat. UltiMate 3000 Pumps LC Columns and Accessories

Accessories for the Use with UltiMate 3000 Pumps For use with SD RS BM XRS BX Description Quantity Cat. No. pumps pumps pumps pumps pumps Filter holder for solvent lter 6 6268.0115 XXX Filter holder for solvent lter 2 6268.0116 X Filter frit (SST, porosity: 10µm) 10 6268.0110 XX Filter frit (Ti, biocompatible, porosity: 10µm) 10 6828.0111 XX X Filter frit (PEEK, porosity: 10µm) 10 6828.0117 X Solvent lines kit with lter frits (SST, porosity: 20µm) 1 6043.8109 X

Spare Parts for UltiMate 3000 Pumps For use with SD RS BM XRS BX

Description Quantity Cat. No. >> pumps pumps pumps pumps pumps

Check Valves LC and LC-MS Instrument Parts Check valve, cartridge (ceramics) for the inlet and outlet valve 1 6041.2301 XXX X Check valve cartridge for the inlet and outlet valve 1 6043.0145 X Pistons Pistons (sapphire) 2 6040.0042 XXX Pistons (ceramics) 2 6040.0842 X Pistons (sapphire) 2 6043.0169 X Piston Seals Piston seal (in plate of seal wash system) Normal Phase 2 6040.0306 XXX Piston seal kit (in plate of seal wash system) Reversed 1 6040.9010 X Phase (2 seals and 1 support ring) Piston seal (main piston seal) Reversed Phase 2 6040.0304 X Piston seal (main piston seal) Reversed Phase 2 6266.0305 X Piston seal kit (main piston seal) Reversed Phase 1 6025.2012 X (2 seals and 1 support ring) Piston seal kit (main piston seal) Reversed Phase 1 6040.9010 X (2 seals and 1 support ring) Piston seal kit (main piston seal) Normal Phase 1 6040.9011 X (2 seals and 1 support ring) Piston seal kit Reversed phase (2 primary piston seals 1 6043.0295 X and 2 secondary piston seals) Support Rings Support ring for pistons seals 2 6040.0012 XX Support ring/pistons seal kit Reversed Phase 1 6025.2012 X (2 piston seals and 1 support ring) Support ring/pistons seal kit Reversed Phase 1 6040.9010 X (2 seals and 1 support ring) Support ring/pistons seal kit Normal phase 1 6040.9011 X (2 seals and 1 support ring)

www.thermoscienti c.com/chromatography 4-213 4 -214 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Normal Phase Kit for SD pumps SD for Kit Phase Normal HPG-3200BX for Kit Phase Normal Description Pumps 3000 UltiMate for Kits Upgrade Phase Normal 1 Pumps 3000 UltiMate with Use the for Systems Mixing syringe 1mL 5mL syringe syringe 100µL valve injection manual the with use 1 for Syringe valve injection manual for kit Mounting kit valve injection manual Motorized MPa >34 pressures biocompatible, kit, valve injection Manual MPa >50 pressures kit, valve injection Manual Description Pumps 3000 UltiMate with Use the for Valves Injection Sample Manual Description ie o 5Lmxn oue1 volume mixing 35µL for Mixer Description Pumps 3000 UltiMate for Kits Maintenance ie o 0µ iigvlm 1 volume mixing 100µL for Mixer aneac i o S-101 ISO-3100 for kit Maintenance ie o 0µ iigvlm 1 1 volume mixing 400µL for Mixer volume mixing 200µL for Mixer Maintenance kit for HPG-3x00 ie o 50Lmxn oue1 1 volume mixing 1550µL for Mixer volume mixing 800µL for Mixer aneac i o P-401 LPG-3400 for kit Maintenance aneac i o G-601 DGP-3600 for kit Maintenance except ISO-3100 except Quantity 1 Quantity SD pumps RS pumps BM pumps XRS pumps XRS pumps BM pumps RS pumps SD 6040.1950 0015 6040.1956A 6040.1953 0015 0015A64.9164.946042.1954 6043.1954 6042.1951 6040.1954A 6040.1951 0015 0015A6042.1952 6040.1955A 6040.1952 SD pumps 0050 6042.5000 6040.5000 0050 6042.5100 6040.5100 1 1 Quantity 1 1 1 1 1 1 Quantity 1 – 6040.5450 6040.5750 6040.5310 6040.5110 Cat. No. Cat. RS pumps 6040.1972 6040.1975 No. Cat. 6040.0620 6035.0670 6035.0665 6040.0611 6040.0110 6040.0610 No. Cat. 6042.0600 6042.1950 – – – – BX pumps – – 6042.1953 – (RP) (NP) UltiMate 3000 Autosamplers LC Columns and Accessories

Sample Racks for the Use with UltiMate 3000 WPS Autosamplers and UltiMate 3000 ACC Autosampler Column Compartments For use with Description Quantity Cat. No. WPS ACC Sample Racks Sample Rack (10 position and/or normal well plates) for 10mL vials, 22.5mm×46mm 1 6820.4086 XX Sample Rack (22 position) for 4mL vials, 15mm×45mm, cylindrical 1 6820.4084 XX Sample Rack (40 position) for 2mL vials, 12mm×32mm, cylindrical 1 6820.4070 XX Sample Rack (40 position and/or normal well plates) for 1.1mL vials, 12mm×32mm, conical 1 6820.4087 XX Sample Rack (40 position) for 0.5mL Eppendorf tubes 1 6820.4096 XX Sample Rack (40 position) for 1.5mL Eppendorf tubes 1 6820.4094 XX Sample Rack (72 position and/or normal well plates) for 1.2mL vials, 8mm×40mm, cylindrical 1 6820.4090 XX Sample Rack (72 position and/or normal well plates) for 0.3mL vials, 8mm OD, cylindrical 1 6820.4091 XX Sample Rack (72 position) for 0.3mL micro dialysis vials 1 6820.4097 XX

Support Racks for Well Plates

Support rack for deep well plates (34 – 46mm high) 1 6820.4079 X >>

Support rack for deep well plates (30 – 36mm high, 96 or 384 wells) 1 6820.4083 X LC and LC-MS Instrument Parts Support rack for deep well plates (20 – 32mm high, 96 or 384 wells) 1 6820.4089 X Find the Thermo Fisher Scienti c Vials and Closures portfolio on page 2-001.

www.thermoscienti c.com/chromatography 4-215 4 -216 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories neto ouekt(00L 1 (1000µL) kit volume Injection neto ouekt(0µ) ie 1 Viper (500µL), kit volume Injection neto ouekt(0µ) ie 1 Viper (500µL), kit volume Injection neto ouekt(5µ) ie 1 Viper (250µL), kit volume Injection Needle (SST) Viper compatible Viper (SST) Needle Needles Description Injection Volume Kits for the Use with UltiMate 3000 WPS-3000 Split Loop Autosamplers Loop Split WPS-3000 3000 UltiMate with Use the for Kits Volume Injection Autosamplers Loop Split WPS-3000 3000 UltiMate with Use the for Accessories Optional neto ouekt(5µ) ie 1 Viper (250µL), kit volume Injection Description edesa ailr,1L ie M3N 1 (MP35N) Viper 1µL, capillary, seat Needle ede(eai)Vprcmail 1 compatible Viper (ceramic) Needle 25µL Sample loop, Viper loop, Sample 25µL 1 -SST Loops 1 Sample (SST) Viper 24µL, capillary, seat Needle (SST) Viper 1µL, capillary, seat Needle edesa ailr,31L ie ST 1 (SST) 3.1µL, Viper capillary, seat Needle Capillaries Seat Needle (SST) Needle (SST) Needle 100µL Sample loop, Viper Viper loop, Sample 100µL (MP35N 1 3.1µL, Viper capillary, seat Needle (PEEK) 3.1µL, Viper capillary, seat Needle 250µL Sample loop, Viper Viper loop, Sample 250µL 500µL Sample loop, Viper loop, Sample 500µL 25µL Sample loop, Viper Viper loop, Sample 25µL –MP35N Loops Sample loop Sample 1000µL 100µL Sample loop, Viper Viper loop, Sample 100µL 250µL Sample loop, Viper loop, Sample 250µL 500µL Sample loop, Viper loop, Sample 500µL Buffer loop, 100µL, Viper (SST) Viper 100µL, loop, Buffer Loops Buffer loop Sample 100µL –PEEK Loops Sample ufrlo,10L ie M3N 1 (MP35N) Viper 100µL, loop, Buffer ufrlo,>20L ie M3N 1 1 (MP35N) Viper >250µL, loop, Buffer (SST) Viper >250µL, loop, Buffer 25µL Syringe Syringe 25µL Syringes 100µL Syringe Syringe 100µL 250µL Syringe Syringe 250µL 500µL Syringe Syringe 500µL 1000µL Syringe Syringe 1000µL 2500µL Syringe 2500µL ™ ) Quantity 1 1 Quantity 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 6822.2436 6841.2443 6822.2443 6841.2442 6822.2442 Cat. No. Cat. 6841.2464 6820.2432 No. Cat. 6841.2472 6841.2420 6820.2452 6820.2409 6820.2472 6820.2464 6820.2419 6820.2403 6820.2451 6827.2408 6820.2453 6820.2454 6841.2452 6820.2429 6841.2451 6841.2453 6841.2454 6820.2466 6820.2431 6841.2466 6841.2468 6820.2468 6822.0001 6822.0002 6822.0003 6822.0004 6822.0005 6822.0006 WPS-3000(T) 3000TXRS/ P-00TR WPS-3000TBRS WPS-3000(T)RS WPS- X/X X/X X/X X/X X/X X/X X/X X/X X/X X/X X/X –/X –/X X/– RS X X 3000TBRS WPS- X X X X X X X X X X X X X X X X X WPS-3000(T) SL Analytical SL For use with use For For use with use For X X X X X X X X X X X X X WPS-3000(T)SL Analytical X X 3000TBSL WPS- X X X X X X X X X X WPS-3000(T)SL WPS-3000(T) Semiprep SL Semiprep SL X X X X X X X X X X X LC Columns and Accessories Optional Accessories for the Use with UltiMate 3000 WPS-3000 Pulled Loop Autosamplers For use with WPS-3000(T)PL WPS-3000TBPL Description Quantity Cat. No. WPS-3000TFC WPS-3000TBFC RS Nano/Cap Analytical Modi cation Kits Nano/Cap modi cation kit 1 6824.0030 X Nano/Cap modi cation kit 1 6825.0030 X Upgrade kit PAEK 1 6821.0045 X Front cover, transparent 1 6820.1427 X X X X Needles 2.4µL Needle, nanoViper 1 6820.3010 X X1 X1 3.6µL Needle, Viper 1 6820.3023 X X X1 X1 15µL Needle, Viper 1 6820.3025 X2 X X X 15µL Needle, nanoViper 1 6820.3115 X2 X X X Sample Loops – nanoViper 1µL Sample loop, nanoViper 1 6826.2401 X X1 X1 5µL Sample loop, nanoViper 1 6826.2405 X X1 X1

10µL Sample loop, nanoViper 1 6826.2410 X X1 X1 >> 20µL Sample loop, nanoViper 1 6826.2420 X X1 X1 LC and LC-MS Instrument Parts 50µL Sample loop, nanoViper2 1 6826.2450 X X X X 125µL Sample loop, nanoViper2 1 6826.2412 X X X X Sample Loops – SST 50µL Sample loop 1 6824.0019 X 250µL Sample loop 1 6824.0020 X Sample Loops – PEEK 5µL Sample loop 1 6823.0016 X X X 10µL Sample loop 1 6823.0017 X X X 20µL Sample loop 1 6823.0018 X X X 50µL Sample loop 1 6823.0019 X2 X X 125µL Sample loop 1 6821.0032 X2 X X 250µL Sample loop 1 6823.0020 X X Buffer Tubings 50 µL Buffer tubing (SST) 1 6820.0019 X X1 50 µL Buffer tubing (PEEK/PTFE) 1 6821.0019 X3 X1 500 µL Buffer tubing (SST) 1 6820.0020 X X 500 µL Buffer tubing (PEEK/PTFE) 1 6821.0020 X3 X X 1000 µL Buffer tubing (SST) 1 6820.0056 X 1000 µL Buffer tubing (PEEK/PTFE) 1 6821.0022 X X 2000 µL Buffer tubing (SST) 1 6820.0021 X 2000 µL Buffer tubing (PEEK/PTFE) 1 6821.0021 X3 Syringes 25µl Syringe 1 6822.0001 X X1 X1 100µl Syringe 1 6822.0002 X X X X 250µl Syringe 1 6822.0003 X4 X X X 500µl Syringe 1 6822.0004 X4 1000µl Syringe 1 6822.0005 X4 Syringe upgrade kit (250µL)4 1 6820.0031 X Syringe upgrade kit (250µL, 1 6821.0031 X3 biocompatible version)4 Bridges 1µL Bridge (PEEKsil) 1 6824.0015 X1 1µL Bridge (PEEKsil) 1 6825.0015 X1 6.2µL Bridge (PEEK) 1 6824.0016 X 6.2µL Bridge (PEEK) 1 6825.0016 X Find recommended combinations for syringes, buffer tubings and sample loops on page 4-218. 1. Can be used for Nano/Cap/Mic applications only if Nano/Cap modi cation kit is previously installed 2. For analytical applications only, please use the smaller sample loops for Nano/Cap 3. Can be mounted only when PAEK Upgrade Kit is previously installed 4. For analytical applications only, please use the smaller syringes for Nano/Cap www.thermoscienti c.com/chromatography 4-217 4 -218 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Front cover, transparent Syringe 1000µL Syringe 250µL Viper loop, Sample 200µL Viper loop, Sample 50µL Viper loop, Sample 20µL Viper >250µL, tubing, Buffer Needle vials 10mL uncapped for pusher vial for Adapter vials 2mL for used be to holder 5-position for Adapter Description Autosampler Column Compartments ACC-3000(T) with Use the for Parts Spare and Accessories tray) per positions (54 vials 2mL for (VT54) tray Sample Syringe 100µL valve injection Cheminert for loop Sample 100µL valve injection Cheminert for loop Sample 20µL valve injection Cheminert for loop Sample 2µL (SST) loop Holding (FEP) kit loop Holding mounted adapter needle with (FEP) loop Holding Description Autosamplers Open OAS-3x00 3000 UltiMate with Use the for Parts Spare and Accessories Syringe Recommended Combinations Autosamplers WPS-3000TBFC and WPS-3000TFC Analytical, WPS-3000(T)BPL Nano/Cap, RS WPS-3000(T)PL 3000 UltiMate with Use the for Combinations Loops Sample Tubings and Buffer Syringes, Recommended 25µL WPS-3000(T)PL RS Nano/Cap 100µL 250µL 100µL WPS-3000TBPL Analytical 250µL 25µL WPS-3000TFC and WPS-3000TBFC Analytical (with Modi Nano/Cap Kit) cation 100μL 250µL 250µL Buffer Tubing Buffer 50µL 500µL 500µL 500μL 1000µL 50µL 500μL 500µL 1000µL 1 1 1 1 1 1 1 1 1 1 Quantity 6 1 1 1 1 1 1 1 Quantity Sample Loop 1µL, 5µL 10µL, 20µL 50µL, 125µL50µL, 5µL, 10µL, 50µL 20µL, 125µL, 250µL 1µL, 5µL 5µL, 10µL, 50µL 20µL, 50µL, 125µL50µL, 250µL 6820.1427 6822.0005 6822.0003 6830.2418 6830.2416 6830.2414 6820.2468 6820.2403 6820.2402 6820.4092 No. Cat. 6845.0055 6845.0062 755.CSL100 755.CSL20 755.CSL2 6845.0029 6845.0035 6845.0034 No. Cat. UltiMate 3000 Thermostatted Column LC Columns and Accessories Compartments

Accessories for the Use with UltiMate 3000 TCC-3000(RS) Thermostatted Column Compartments Description Quantity Cat. No. 1µL Pre-column heater, Viper (SST) 1 6722.0510 2µL Pre-column heater, Viper (SST) 1 6722.0530 7µL Pre-column heater, Viper (SST) 1 6722.0540 11µL Pre-column heater, Viper (SST) 1 6722.0550 Post-column cooler insert, Viper (SST) 1 6730.0008 Column clip for columns with an outer diameter < 8mm 6 6722.0290 Column clip for columns with an outer diameter of 8-12mm 6 6722.0280

Optional Column Switching Valves for the Use with UltiMate 3000 TCC-3000RS/SD Thermostatted Column Compartments >>

Biocompatible Non biocompatible LC and LC-MS Instrument Parts TitanHP™ TitanHT™ TitanHP™ TitanHT™ < 34 MPa < 41 MPa < 103 MPa < 41 MPa < 103 MPa <125 MPa Description Quantity (5000 psi) (6000 psi) (15000 psi) (6000 psi) (15000 psi) (18130 psi) Valve Drives Valve Drive (Actuator) 1 6730.0003 6730.0001 6730.0003 6730.0001 for right side installation Valve Drive (Actuator) 1 6730.0004 6730.0002 6730.0004 6730.0002 for left side installation Pods Pod, 2-position, 6-port 1 6723.9013 – 6730.0031 6722.9013 6730.0006 – (PEEK) (Ti) (SST) (SST) Pod, 2-position, 10-port 1 6723.9023 – 6730.0032 6722.9023 6730.0026 – (PEEK) (Ti) (SST) (SST) Pod, 6-position, 7-port 1 – 6722.9035 6730.0030 – 6730.0016 6730.0050 (Ti) (Ti) (SST) (SST)

www.thermoscienti c.com/chromatography 4-219 4 -220 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories UltiMate 3000 Detectors 3000 UltiMate Description Detectors Optical 3000 UltiMate for Parts Spare and Accessories Description Detectors FLD-3x00(RS) and VWD-3x00(RS) MWD-3000(RS), DAD-3000(RS), 3000 UltiMate for Cells Flow DAC cable (analog out) (analog cable DAC DAC board Tungsten lamp lamp Deuterium SST,4MPa) (2µL, micro cell, Flow SST,2MPa) (8µL, analytical cell, Flow 1 dummy cell, Flow MPa) 30 silica, fused (45nL, dispersion ultra-low monitor, UV MPa) 20 silica, fused (3nL, nano cell, Flow MPa) 20 silica, fused (45nL, capillary cell, Flow MPa) 20 silica, fused (180nL, Mic cell, Flow 10 MPa) PEEK, (0.7µL, semi-preparative cell, Flow 5MPa) PEEK, (2.5µL, semi-micro cell, Flow 5MPa) (11µL, PEEK, analytical cell, Flow SST,12 MPa) (2.5µL, semi-micro cell, Flow (11µL, SST,12 MPa) analytical cell, Flow  10 MPa) PEEK, (0.7µL, cell Semipreparative ow 5MPa)  PEEK, Semi-micro (2.5µL, cell ow  5MPa) PEEK, Analytical (13µL, cell ow  SST,12 MPa) Semi-micro (2.5µL, cell ow  SST,12 MPa) (5µL, cell Semi-analytical ow  SST,12Analytical MPa) (13µL, cell ow Quantity Quantity 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Cat. No. Cat. No. Cat. 6074.0002 6082.0305 6074.2000 6074.1110 6078.4330 6078.4230 6074.0190 6074.0285 6074.0270 6074.0280 6074.0290 6074.0320 6074.0300 6074.0200 6074.0360 6074.0250 6082.0600 6082.0500 6082.0400 6082.0300 6082.0200 6082.0100 MWD-3000(RS) MWD-3000(RS) DAD-3000(RS) DAD-3000(RS) X X X X X X X X X X VWD-3x00(RS) VWD-3x00(RS) X X X X X X X X X X X X X X FLD-3x00(RS) FLD-3x00(RS) X X X LC Columns and Accessories Accessories and Spare Parts for the Use with the UltiMate 3000 PCM-3000 pH and Conductivity Monitor Description Quantity Cat. No. pH  ow cell, 28µL (PEEK) 1 6082.2030 pH electrode for PCM-3000 1 6082.2020 Dummy electrode for PCM-3000 1 6082.2042 Conductivity  ow cell, 21µL (Ti) 1 6082.2060

Accessories and Spare Parts for the Use with the UltiMate 3000 ECD-3000RS Detectors Description Quantity Cat. No. In-Line Filter Kit with Graphite Filter Elements 1 70-0893 In-Line Filter Kit with PEEK Filter Elements 1 70-4093 Filter Element (Graphite) 5 70-0898 Filter Element (PEEK) 5 70-3824

Amperometric Cell 6041RS ultra (25nL or 50nL, 1 6070.3000 glassy carbon or boron-doped diamond, 1.4 MPa) >>

Gasket for Amperometric Cell 6041RS (25nL, boPET) 5 6070.2528 LC and LC-MS Instrument Parts Gasket for Amperometric Cell 6041RS (50nL, boPET) 5 6070.2529 Working Electrode for Amperometric Cell 6041RS 1 6070.3100 (boron-doped diamond, BDD) Working Electrode for Amperometric Cell 6041RS 1 6070.3200 (glassy carbon, GC) Coulometric Cell 6011RS ultra 1 6070.2400 (7.06µL, micro-porous graphite carbon, 4 MPa) Potentiostat Module, dual channel DC 1 6070.1400 Cell Simulator – SimulatorRS 1 6070.4100 Cell Simulator – Quali erRS 1 6070.4200

Accessories and Spare Parts for the Use with Charged Aerosol Detectors Description Quantity Cat. No. Corona CAD Corona ultra RS Gas Filter Assembly (HEPA/Carbon) 1 70-6224 X X Corona Waste Bottle Cap Assembly 1 70-7754 X X Bottle, 5.0 L Waste (Compatible with 70-7754 only) 1 70-7751 X X Filter-HPLC Inline 1 70-4538 X X Drain/Vent Tubing Assembly 1 70-7115 X X Gas Inlet Tubing (¼") 1 70-6260 X X Gas Exhaust Hose Assembly 1 70-6261 X X Maintenance Kit for Nitrogen Generator 1 70-6230 X X Corona ultra RS Waste Level Detector Assembly 1 70-9363R X

www.thermoscienti c.com/chromatography 4-221 4 -222 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories UltiMate 3000 Automated Fraction Collector Fraction Automated 3000 UltiMate Find the Vials and Closures portfolio on page 2 page on portfolio Closures and Vials the Find Description AFC-3000 3000 UltiMate the with Use the for Accessories and Racks Sample Sample Rack (10 position) for 10mL vials, 22mm ×45mm 22mm vials, 10mL for (10 position) Rack Sample Racks Sample Sample Rack (22 position) for 4mL vials, 15mm ×45mm 15mm vials, 4mL for position) (22 Rack Sample Sample Rack (40 position) vfr 2mL vials, 12mm×32mm vials, 2mL vfr position) (40 Rack Sample Sample Rack (21 position) for 50mL tubes, 30mm ×100mm 30mm tubes, 50mL for position) (21 Rack Sample Sample Rack (24 position) for 30mL tubes, 24mm ×100mm 24mm tubes, 30mL for position) (24 Rack Sample Sample Rack (40 position) for 20mL tubes, 20mm ×100mm 20mm tubes, 20mL for position) (40 Rack Sample Sample Rack (60 position) for 14mL tubes, 16mm ×100mm 16mm tubes, 14mL for position) (60 Rack Sample Sample Rack (90 position) for 8mL tubes, 13mm ×100mm 13mm tubes, 8mL for position) (90 Rack Sample apeRc 3 oiin o oyvas cnilto il,2m D1 OD 28mm vials, scintillation vials, Foxy for position) (36 Rack Sample Sample Rack (60 position) for Foxy vials, 1.5mL vials, Foxy for position) (60 Rack Sample Sample Rack (72 position) for Foxy mini tubes, 18mm OD 18mm tubes, mini Foxy for position) (72 Rack Sample Positioning pins for Foxy racks Foxy for pins Positioning Funnel rack Adapter for 4 WPS racks 4WPS for Adapter Adapter for 4 well plates 4well for Adapter Adapter for 2 well plates (Foxy Jr./R1) (Foxy plates 2well for Adapter (“Kit for low  low for (“Kit rates”) ow tubing and former, drop ID 0.4mm valve, Diverter Accessories (“Kit for normal-phase LC”) normal-phase for (“Kit cable grounding 1mm SST,including ID former, Drop -001 . Quantity 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 Cat. No. Cat. 6820.4086 6820.4084 6820.4070 6702.0021 6702.0024 6702.0040 6702.0060 6702.0090 5701.2025 5701.2023 5701.2024 6702.9006 6702.1021 6702.0100 6702.0200 5701.2021 6702.0300 6702.0400 UltiMate 3000 RSLC Nano System LC Columns and Accessories

The UltiMate 3000 RSLCnano System was developed with throughput in mind. The robust, continuous direct-flow delivery is designed for interruption-free analysis. The wide flowpressure footprint enables the application of UHPLC to the nano scale, allowing analysts to tune for the highest resolution or the fastest analysis time, even for tryptic peptide samples of utmost complexity. Configurable for speed, separation power, or sensitivity, the UltiMate 3000 RSLCnano is the only system to deliver all.

Consumables and Spare Parts for UltiMate 3000 RSLC Nano Systems Description Quantity Cat. No. Maintenance kit for NCS-3500RS loading pump 1 6042.1951 Check valve cartridge, ceramics (for both the inlet and outlet valve) for the NC pump and loading pump 1 6041.2301 Column switching valve; 2-position, 6-port valve 1 6041.0004 Column switching valve; 2-position, 10-port valve 1 6041.0001 Flow meter, biocompatible with  ow selector for nano LC (50 – 1000nL/min) 1 6041.7901A

Flow meter, biocompatible with  ow selector for capillary LC (0.5 – 10µL/min) 1 6041.7902A >> Flow meter, biocompatible with  ow selector for micro LC (5 – 50µL/min) 1 6041.7903A LC and LC-MS Instrument Parts Flow selector for nano LC (50 – 1000nL/min) 1 6041.0002 Flow selector for capillary LC (0.5 – 10µL/min) 1 6041.0003 Flow selector for micro LC (5 – 50µL/min) 1 6041.0014 Mixer kit (8µL) for capillary LC 1 6041.7130 Piston (sapphire) 2 6040.0042 Piston seal (in plate of seal wash system), NP, for NC pump and loading pump 1 6040.0306 Piston seal (main piston seal), RP, for NC pump and loading pump 2 6266.0305 Support ring for main piston seal for NC pump and loading pump 2 6040.0012 Pump head, entire assembly, for NC pump 1 6041.1901A Pump head, entire assembly, for loading pump 1 6041.1902 Rear seal wash system, seals 1 6040.2208 Column installation/capillary clips kit (6 column clips, 2 clips to attach capillaries routed 1 6041.0011 from top to bottom through the column chamber) Tubing from proportioning valve to pump head, loading pump (with ttings) 2 6040.3023

EASY-nLC 1000 Liquid Chromatograph

Thermo Scienti c™ EASY-nLC 1000 Liquid Chromatograph provides effortless split-free, nano- ow UHPLC performance up to 1000 bar with minimal investment. Dual in-line  ow sensors before mixing gives uncompromising gradient precision. Identify more peptides and increase productivity with higher pressure without compromising reliability and robustness.

Common replacement parts for EASY-nLC 1000 Systems Description Cat. No. Pump Piston Seal Replacement Kit (contains four spring-energized piston LC510 seals and a piston seal tool) Valve rotor seal (contains one rotor seal) LC228 Autosampler needle, ASC model LC302 Column Out solvent line LC560 Waste In solvent line LC562 Flow sensor lters (contains four  ow sensor lters) LC542

www.thermoscienti c.com/chromatography 4-223 4 -224 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Thermo Scienti Thermo c Accela High Speed LC Accessories, Consumables for and Spare Parts Tubing assembly, 2.5mL syringe 2.5mL Tubing assembly, valve injection Valco Rotor, Rotor Seal, injector Valve, Stripper 1000µL Loop, Sample 500µL Loop, Sample 100µL Loop, Sample 50µL Loop, Sample 25µL Loop, Sample 20µL Loop, Sample 10µL Loop, Sample 5µL Loop, Sample 2.5mL Syringe, ext) tubing needle (with 2500µL standard, Syringe, port) (injection port needle Retainer, Reservoir vials, 16mL Plunger, 500µL concentric, replacement, Plunger, 250µL concentric, replacement, Plunger, 100µL concentric, replacement, 500µL concentric, Syringe, 250µL concentric, Syringe, 100µL concentric, Syringe, needlePort, 1/16in tube OD 10-32, long, compression, Nut, Lubricant, Triow Swagelok Back, Ferrule Swagelok Front, Ferrule bottle wash Kit, tubing needle Kit, Kit, maintenance Accela Autosampler Silicon/Te screw) Grease, lead (for on Filter, solvent ush well 96 Adapter, Cooling ID 0.012in tube, transfer Assembly, Assembly, syringe valve kit bottle wash Assembly, tubing needle Assembly, inert needle, Assembly, Description Consumables Autosampler Accela Accela PDA Detector accessory kit Kit Accessory Autosampler Accela Pump Accela Seal, Kit, Pump Accela maintenance, Kit, kit interconnect solvent system LC/MS Accela Pump accessory kit Description Kits Accela Accessory 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Quantity 1 1 1 1 1 1 Quantity 60053-60005 00110-03-00019 00109-99-00021 00110-03-00013 F1034-010 00109-99-00014 00109-99-00013 00109-99-00012 00109-99-00011 00109-99-00010 00109-99-00009 00109-99-00008 00109-99-00007 60053-60006 60053-62002 60053-10035 00301-07527 F1123-030 F1123-020 F1123-010 F1100-030 F1100-020 F1100-010 60053-20031 00101-07-00001 1611-0030 00101-08-00002 00101-08-00001 60053-62009 60053-62043 60357-62002 00301-01910 A4258-010 60053-20002 60053-60014 A3692-010 60053-60041 60053-60102 60357-60017 No. Cat. 60257-62001 60357-62001 60157-62003 60157-62002 F5050-010 60157-62001 No. Cat. LC Columns and Accessories Accela Open Autosampler Consumables Kit Description Cat. No. Kit, asp tube, Accela Open A/S, RoHS PAL DilAspKit 00950-01-00311 Loop, hldg, dlw, Accela Open A/S, RoHS PAL DLWLoop 00950-01-00317 Screw, syringe, Accela Open A/S, RoHS PAL DLW Screw 00950-01-00322 Tool, needle, Accela Open A/S, RoHS PAL DLW NdTool 00950-01-00323 Holder, needle, dlw, Accela Open A/S, RoHS PAL DLWNHA 00950-01-00324 Syringe, dlw, Accela Open A/S, RoHS SyrC DLW 100-R 00950-01-00325 Plunger, syringe, dlw, Accela Open A/S, RoHS PLG DLW 100 00950-01-00326 Holder, needle, dlw, Accela Open A/S, RoHS PAL DLWPlg 00950-01-00327 Needle kit, dlw, 3PK, Accela Open A/S, RoHS PAL DLWNdl 00950-01-00328 Kit, dlw replament, Accela Open A/S, RoHS PAL DLW Option 00950-01-00330 Kit, insert, dlw, Accela Open A/S, RoHS PAL DLW Insert 00950-01-00334 Tube, waste, dlw, Accela Open A/S, RoHS PAL TubeWaste 00950-01-00335 Valve, inject, 18K, Accela Open A/S, RoH N/A 00950-01-00336 Rotor, Inj Valve, VALCO 18KPSI, RoHS C72-16R6 00110-03-00019 Stator Inj Valve, VALCO 18KPSI, RoHS C72V-6C96 00950-01-00337

Accela PDA Accessories >>

Description Quantity Cat. No. LC and LC-MS Instrument Parts Filter Wheel for linearity calibration, (5 position; 1 cuvette with 1 803264 perchloric acid blank and 4 cuvettes with different concentrations of potassium dichromate in perchloric acid solution, NIST traceable) 10mm  owcell assembly, with inlet/outlet tubing and ttings (1cm LightPipe) 1 803265-01 Flowcell Assembly, with inlet/outlet tubing and ttings (5cm Lightpipe) 1 803237-01 Backpressure regulator 1 802259 FingerTight PEEK Ferrule Nuts 1 2522-0285 Inlet tubing, with insulation, PEEK 1/16 x 0.005in ID (red) 1 803260 Outlet tubing, PEEK 1/16 x 0.01in ID (blue) 1 703950

Accela Pump Liquid Displacement Assembly Parts Description Material Quantity Cat.No. Seal, lm, PEEK, Accela pump PEEK 4 00950-01-00122 Seal, high pressure, GFP, Accela pump PTFE/GFP/Ti 2 00950-01-00129 Valve, check, outlet, Accela pump Ti/ruby/sapphire 1 00950-01-00131 Valve, check, inlet, Accela pump Ti/ruby/sapphire 1 00950-01-00130 Cartridge, blind, inlet, Accela pump PEEK 1 00950-01-00120 Piston, sapphire Sapphire 2 00950-01-00126 Secondary piston seal (wash seal) PE/SS 2 00950-01-00128

Accela 1250 Pump Liquid Displacement Assembly Description Quantity Cat. No. UHP PEEK sealing ring 5 00950-01-00284 Check valve, Titanium and ruby 2 00110-05110 Piston unit, sapphire 2 00950-01-00126 Secondary piston seal (GFP) 2 00950-01-00129 Primary piston seal (GFP) 2 00950-01-00129

Accela 600 Pump Spare Parts Description Quantity Cat. No. O-ring, 2.69mm x 1.14mm, per uor 1 00950-01-00286 Kit, inlet tubing, Accela 600 pump 1 60157-62008 Mixer, static, 65µL, Accela 600 pump, RoHS 1 00950-01-00292 Seal, H/P 2 00107-18110 Piston, 1/8", TZP 2 00201-11324 Valve, check 2 00110-05110 Seal, face, PEEK, check valve 5 00950-01-10013 Seal, piston, secondary 2 00107-18114

www.thermoscienti c.com/chromatography 4-225 4 -226 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories for high-resolution,accurate-massmeasurements. accommodate complexmatricesencounteredinavarietyofapplications. LC-MS, ourmassspectrometerscoupleperfectlywithHPLC,UHPLC,andnano-LCs toeasily in complexbiologicalmatrices. identification androutineHPLCdetectionthroughthemostdemandinganalysesof low-levelcomponents identification oflow-levelanalytes.Iontrapsystemssupportavarietyapplications fromcompound dependent analysisalongwithexcellentfull-scansensitivitytoprovideroutine detection andrapid and methodologies. The systemcanbeusedwithbothHPLCandICsystemsinawiderange ofapplications price andspacerequirements. Our singlequadrupoleLC-MSsystemofferssuperiorease-of-use andmodest Plus SingleMSQ Quadrupole LC-MS value-conscious ThermoScientific TSQ Seriestriple-stagequadrupoleLC-MSsystemsofferawiderangeofcapabilities. Fromthe quantitation oftargetcompoundsincomplexmatrices. Robust triple-stagequadrupoleLC-MSsystemsenableultra-sensitive Triple Series TSQ Quadrupole LC-MS application andbudget.IontrapLC-MSsystemsofferuniquecapabilitiessuchas MS With marketandtechnologyleadershipforover25years,weofferiontrapLC-MS systemsforevery capable ofprovidingallfourbenefitsatthesametime,withoutcompromise. superior dynamicrangeandunsurpassedsensitivity, Orbitrapsystemsaretheonlymassspectrometers most confidentproteinandmetaboliteidentification,characterizationquantitation. Combinedwith to reduceanalysistimesandincreaseconfidenceinresults.Thismakesittheplatform ofchoiceforthe Orbitrap LC-MStechnologyroutinelydeliversthesuperiormassresolutionand mass accuracynecessary Thermo Scientific Ion Traps Series and Orbitrap Scienti Thermo Systems c LC-MS Accessories, Consumables for and Spare Parts ™ Orbitrap ™ ™ LC-MSsystemsaretherecognizedstandards TSQ Endura ™ LC-MStothepreciseThermoScientific n anddata- ™ TSQQuantiva ™

Ion Traps and Orbitrap Series LC Columns and Accessories

API Source Components Fittings, Ferrules, Sample Loops and Tubing Description Cat. No. Description Cat. No. USI ESI probe OPTON-20011 Fitting, HPLC, adaptor 10-32 x 1/4 PEEK 00101-18080 USI APCI probe OPTON-20012 Fitting, nut, ngertight, HPLC, 10-32, PEEK 00101-18081 Capillary heater cage assembly, for LTQ & LTQ XL 97055-60040 Fitting, ngertight, 2, Upchurch 00101-18195 Capillary heater cage assembly, for the LXQ & LCQ Fleet 97055-60181 Fitting,  angeless, 1/8in, Delrin, green 00101-18198 Fitting,  angeless, 1/8in, Delrin, blue 00101-18200 Fitting, plug, 1/4-28, Tefzel 00101-18075 Accessory Kits Nut, LC, 1/16in, ss, Rheodyne 2522-0066 Description Cat. No. Ferrule, LC, 1/16in, ss, Rheodyne 2522-3830 TSQ Quantum forepump kit (used with LTQ series) 70111-62014 Ferrule, 0.008in ID, Kel-F HPLC 00101-18114 Hose and accessories kit, LTQ & LTQ XL 97055-62007 Ferrule, 0.012in ID, Kel-F HPLC 00101-18116 Hose, single mechanical pump accessory, LXQ & LCQ Fleet 97055-60135 Ferrule, 0.016in ID, PEEK, HPLC 00101-18120 LTQ series ship kit 70111-62033 Ferrule, Fingertight 2, Upchurch 00101-18196 Accessory Kit, LTQ/LTQ XL 97055-62044 Ferrule, 1/8in, Tefzel 00101-18199 >> Accessory Kit, LXQ & LCQ Fleet 97055-62045 Fitting, grounding union, 1/16, ss 00101-18182 Fitting, LC union, 0.010in ori ce, PEEK 00101-18202 LC and LC-MS Instrument Parts Fitting, LC TEE union, 0.020in ori ce, PEEK 00101-18204 Chemicals 5µL sample loop, ss 00110-22026 Description Cat. No. 10µL sample loop, ss 00110-22012 LCQ chemicals kit, for LTQ & LCQ series 97000-62042 20µL sample loop, ss 00110-22028 Met-Arg-Phe-Ala, 20mg 00301-07709 50µL sample loop, ss 207180 Ultramark 1621 00301-12200 100µL sample loop, ss 00110-22018 Caffeine, 1mg/mL 00301-12310 500µL sample loop, ss 00110-22020 Reserpine, 1g 00301-12901 1mL sample loop, ss 00110-22022 Tubing, 0.15mm ID x 0.39mm OD fused-silica capillary 00106-10498 for APCI sample tube Pressure Gauges Tubing, 0.10mm ID x 0.19mm OD fused-silica capillary 00106-10499 Description Cat. No. for ESI sample tube Ion gauge 00105-01525 Tubing, 0.05mm ID x 0.19mm OD fused-silica capillary 00106-10502 for ESI sample tube for low  ow up to 200µL/min Ion gauge o-ring, 0.737id 3/32 THK Viton 00107-10056 Tubing, 0.1mm ID x 0.363mm OD fused-silica capillary 00106-10504 for infusion line Metal Needle Kits Tubing, 0.075mm ID x 0.3193mm OD fused-silica capillary 00106-10511 for low  ow applications instead of metal needle Description Cat. No. Te on tube, .03in ID x 1/16in OD for syringe adapter 00301-22915 32-Gauge metal needle kit for high  ow LC  ow rates OPTON-53003 assembly between 5µL/min to 400µL/min Uses 32-gauge Tubing, 1in ID x 3/16in, Tygon 00301-22922 needle, (P/N 00950-00954) Hose, 1.5in ID, PVC reinforced 00301-24163 34-Gauge metal needle kit for low  ow LC  ow rates OPTON-30004 between 500nL/min to 10µL/min Uses 34-gauge needle, (P/N 97144-20040)

www.thermoscienti c.com/chromatography 4-227 4 -228 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories TSQ Series Triple Series TSQ Quadrupole LC-MS Description Kits Needle Metal Description O-rings and Seals Tubes, Transfer Ion Description Chemicals Endura) TSQ and Quantiva (TSQ Flow Standard Insert Needle Metal Contains Metal Needle PN 70005-20434 PN Needle Metal Contains 2000μL/min to 5μL/min between rates Flow LC 1 Insert Needle Metal 32-Gauge Endura) TSQ and Quantiva (TSQ Flow Low Insert Needle Metal Contains Metal Needle PN 70005-20483 PN Needle Metal Contains 10μL/min to 1μL/min between rates Flow LC Insert Needle Metal 34-Gauge High High kit needle metal ow Low Low kit needle metal ow High High kit needle metal ow Low Low kit needle metal ow High High kit needle metal ow Low Low kit needle metal ow H-ESI metal needle kit needle metal H-ESI TSQ Quantiva iodide Cesium TSQ Endura TSQ kit calibration AM Quantum TSQ TSQ Quantum Access Access Quantum TSQ Reserpine TSQ Quantum Ultra, Ultra AM & Ultra EMR EMR &Ultra AM Ultra Ultra, Quantum TSQ (liquid) solution calibration Polytyrosine-1,3,6 TSQ Quantum Discovery Max Max Discovery Quantum TSQ (solid) standard calibration Polytyrosine-1,3,6 S unu,QatmA unu icvr 1 Discovery &Quantum AM Quantum Quantum, TSQ liquid calibration FC43 TSQ Vantage, Vantage AM & Vantage EMR &Vantage AM Vantage Vantage, TSQ calibrant of infusion for Kit Adapter Syringe Seal for ion transfer tube (TSQ Quantiva) (TSQ tube transfer ion for Seal Graphite vespel o-ring (not TSQ Quantiva) Quantiva) TSQ (not o-ring vespel Graphite Quantity Quantity 1 1 x1g vial 1 1 1 1 x1g vial 1 1 x20mL 1 1 1 vial 1 1 1 1 1 Quantity 1 1 1 1 1 1 1 1 1 1 Cat. No. Cat. 80000-60152 80000-60317 OPTON-53010 OPTON-53011 OPTON-20004 OPTON-20005 OPTON-20016 OPTON-20017 OPTON-53003 OPTON-30004 OPTON-20034 Cat. No. Cat. No. Cat. 80100-20641 HAZMAT-01-00004 70005-20606 70111-62029S 70111-20972 00301-12901 97055-20199 00301-22924 70111-20396 00301-22925 70111-20100 50010-30059 70005-20423 70005-62011 70005-20922 97055-20442 LC Columns and Accessories MS Maintenance Kit Description Quantity Cat. No. MS maintenance kit 1 70111-62032 Ferrule, 0.008 ID, Kel-F (for fused silica sample tube) 2 00101-18114 Tubing, fused-silica, 0.10 ID x 0.19 OD (fused silica sample tube) 6ft 00106-10499 O-ring, Viton, 0.125 ID x 1/16 (for source mount assembly gas connections) 2 00107-02550 Fitting, HPLC, 10-32, short one-piece, 10/pk, RoHS 3 00109-99-00016 Oil, vacuum pump, 1 liter 3 00301-15101 Syringe, 500µL, Gastight 1 00301-19016 Tubing, PEEK, 0.005in ID x 1/16in OD, red 3ft 00301-22912 Tube, Te on, 0.03in ID x 1/16in OD (for syringe adapter) 0.1ft 00301-22915 Polytyrosine-1,3,6 calibration standard (liquid) 1 00301-22924 Screw, 6-32 x 3/8 (used to secure front panels to chassis) 2 00405-63266 Screw, 6-32 x 5/16 (used to secure side panels and EMI shield to chassis) 2 00407-63205 Screw, 4 x 6mm (used to secure PS2 and PS3 to chassis) 2 00407-90000 Screw, 8-32 x 3/8 (used to secure top cover to chassis) 2 00415-83206 O-ring, graphite Vespel (for ion transfer tube) 1 97055-20442

>> LC and LC-MS Instrument Parts ESI Needles and Needle Seal Description Quantity Cat. No. ESI needle 1 00950-00990 ESI needle seal 1 00950-00952 H-ESI needle 1 97055-20273 H-ESI needle seal 1 97055-20271

Viper Fittings (for grounded union to sprayer/probe) Description Cat. No. Viper Capillary, 65 µm x 150 mm PEEK 6041.5615 Viper Capillary, 130 µm x 150 mm PEEK 6041.5616 Viper Kit, 65 µm x 150mm PEEK (5 capillaries) 6041.5601 Viper Kit, 130 µm x 150mm PEEK (5 capillaries) 6041.5602

Pump Oil Description Cat. No. Pump oil for Sogevac sv65 mechanical pump HAZMAT-01-00063

www.thermoscienti c.com/chromatography 4-229 4 -230 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories * * The ConsumablesKitcontainsthenon-liquidpartsrequiredforupkeepofyourMSQPlusMSDetector including MSQ Plus SingleMSQ Quadrupole LC-MS source O-rings,entrancecones,esi/apcicapillaries,conewashassembly, esiproberepairsupplies,turbooilwick Validation MSQ Plus on Xcalibur 2.0 – MS only –MS 2.0 Xcalibur on Plus MSQ Validation 2.02 Devices LC includes 2.0 Xcalibur with 2.0 MSQ to Upgrade Description Software needle pin corona APCI 2m cable USB Hexapole screws – 3 required 1in blue, ID hose, exhaust Vacuum assembly heater Probe complete probe APCI ESI probe complete oil pump vacuum Edwards oil wick replacement turbo Pfeiffer Te6mm supply nitrogen for tubing on nitrogen x1/4in NPT 6mm tting for 6mm tubing nitrogen connector Swage cone (Extraction) Exit O-ring spare cone Entrance O-ring nozzle wash Cone assembly nozzle wash Cone O-ring with complete cone entrance Titanium 1.0 version classic MSQ solution Calibration higher 1.4 and version MSQ solution Calibration sleeve ceramic ESI ferrule 1/16in Vespel Graphite probe API For insert tube PEEK ESI probe capillary capillary probe APCI supplies: repair Probe Description Spares General and Consumables Test Kit MSQ Surveyor te –insulating Kit Repair parts on Heater Probe Kit* Maintenance Annual Description Kits Plus MSQ 1 1 1 Quantity 1 1 1 Per foot 1 1 1 liter 1 1 Per foot 1 1 1 (10) 1pk (10) 1pk 1 1 1 1 1 (10)1 pk (12)1 pk 1 1 Quantity OPTON-09015 OPTON-20432 No. Cat. FM104284 60111-62010 60111-62014 No. Cat. 70005-98033 00302-99-00008 60111-20055 00301-08301 60111-60023 FM102587 FM102595 00301-15102 00950-01116 00109-99-00004 00103-02-00001 00101-02-00006 FM102263 5711000 FM101464 FM102521 60111-60049 FM104285 60111-01001 FM103394 6070119 FM102591 FM102598 FM102594 No. Cat. LC Technical Information LC Columns and Accessories

Method Transfer to Accucore 2.6 μm Columns

Containing solid core particles, which are 1. Scale the Flow Rate 3. Adjust the Gradient Pro le engineered to a diameter of 2.6μm and a very To maintain an equivalent separation when Geometrical transfer of the gradient requires narrow particle size distribution; Accucore transferring a method it is important to keep calculation of the number of column volumes HPLC columns allows high speed, high the linear velocity constant between the original of mobile phase in each segment (time interval) resolution separation, with back pressures and new method. The linear velocity is related of the gradient in the original method to ensure significantly lower than those associated with to the flow rate, internal diameter of the column that the new calculated gradient takes place over sub-2μm fully porous particles. and particle size. A simple equation can be the same number of column volumes, for the

When transferring methods from conventional derived to calculate the flow rate (F2) required new column. HPLC to a solid core column several approaches for the new method. This is shown below, The following calculation should be performed can be taken, depending on the analytical needs. normalized for particle size. for each time segment of the gradient, including If column dimensions are maintained and the 2 2 F2 = F1 x (dc2 / dc1 ) x (dp1 / dp2) column re-equilibration. It takes into consideration particle characteristics, design and diameter, are the void volume of each column (Vc, calculation F1 – original flow rate (mL/min) changed then an improvement in efficiency and, described below), the flow rate in the original >> dc1 – original column internal diameter (mm) therefore, sensitivity, resolution and peak capacity method and the flow rate in the new method Technical Information dp1 – original column particle size (µm) is obtained. A second approach is to change not (calculated in step 1 above) and the time segment dc2 – new column internal diameter (mm) only particle characteristics but also column in the original method. dp2 – new column particle size (µm) dimensions, which has the benefit of further t = t x (V /V ) x (F /F ) reducing analysis time and increasing sensitivity. 2. Adjust the Injection Volume g2 g1 c2 c1 1 2 t – Time segment in original gradient (min) Because sub-3μm solid core based methods g1 An understanding of some practical calculations t – Time segment in new gradient (min) are most often transferred to smaller volume g2 can help to achieve the correct scaling and maintain V – Original column void volume (mL) columns, the same injection volume will take c1 a consistent assay profile between the original V – New column void volume (mL) up a larger proportion of the new column, c2 and transferred method. An identical approach F – Original flow rate (mL/min) possibly leading to band broadening or 1 can be used when transferring a conventional F – New flow rate (mL/min) HPLC analysis to a UHPLC method using potentially overloading the column. It is 2 The void volume of the column is the volume columns packed with sub-2μm fully porous therefore important to scale down the injection that is not taken up by the stationary phase particles such as Hypersil GOLD 1.9μm and volume to match the change in column volume. (approximately 68% of the column volume): Syncronis 1.7μm. Once again, a simple equation can be used to 2 calculate the injection volume (Vi2) required for V = 0.68 x π x r x L There are three main considerations when c the new method. transferring a method to a shorter column Vc – column volume (mL) V = V x (d 2 x L / d 2 x L ) using different particles: Scaling the flow rate, i2 i1 c2 2 c1 1 L – column length (cm) r – column radius (cm) adjusting the injection volume and adjusting Vi1 – original injection volume (µL) the gradient profile. These are discussed in dc1 – original column internal diameter (mm) An example of a method transferred following more detail below.* L1 – original column length (mm) steps 1 to 3 above is illustrated in the

Vi2 – new injection volume (µL) following table:

dc2 – new column internal diameter (mm)

L2 – new column length (mm)

www.thermoscienti c.com/chromatography 4-231 4 -232 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories results innarrowchromatographicpeaks can beoptimizedtogivefastanalysis.This With 1.9µmparticles,operatingparameters Detector SamplingRate column dispersion. making itevenmorecriticaltominimizeextra peaks whichhavealowerpeakvolume smaller columnvolumesandforlessretained for scaleddownseparationsbecauseofthe extra columneffectsbecomemoresignificant detector andintheflowcell.The tubing connectingthecolumntoinjectorand arise fromthecolumn,autosampler, the the chromatographicperformance.Thiscan broadening, whichhasadetrimentaleffecton Excess systemvolumegivesrisetoband Minimizing SystemVolume To obtainthebestdatausingfastchro System Considerations Resource Center (www.thermoscienti c.com/crc) Chromatography the at calculator transfer method aconvenient offer *We columns). x2.1mm, 2.6μm 50 and x2.1mm, (100 2.6μm Accucore to columns) 5μm x4.6mm, (150 HPLC from conditions transfer Method it iscriticalthattheLCinstrument minimal. gradients pumpdwellvolumeneedstobe carefully selected,andwhenrunningfast time constantandsamplingrateneedtobe cell volumeinUV)mustbeminimized,detector tubing IDandlength,injectionvolume,flow be considered.Systemvolume(connecting All systemcomponentsfortheassayshould optimized tooperateundertheseconditions. Solvent Used(mL) Solvent Run Time (minutes) Time Run Resolution Backpressure 17.0 11.0 10.0 0.0 Gradient Time (min) Time Gradient Inj. volume (µL) volume Inj. Flow rate (mL/min) rate Flow Column: Fully porous 5 µm, 150 x4.6mm 150 5µm, porous Fully systemis matography matography 17 17.0 2.6 59 506.0 35.0 35.0 60.0 500.0 35.0 %B 1 1.00 of theanalytes. hold attheendofgradienttoallowelution become necessarytointroduceanisocratic rapid gradientsthisistooslowanditmay get thegradienttocolumn.Whenrunning dwell volumeof800µLwilltake2minutesto will get immediately. Apumpwitha65µLdwellvolume theoretical gradientreachesthecolumn 0.4mL/min andafastgradientof1minute,the we consideramethodusingflowrateof gradient toreachtheheadofcolumn.If dwell volumeaffectsthetimeittakesfor packed columns.Thisisbecausethepump high throughputseparationsonsmallparticle applications usingfastgradients,typicalof important whenrunninghighspeed The HPLCpumpdwellvolumeisparticularly Dwell Volume will becompromised. resolution, efficiencyandanalytical seconds. to achieveoptimumpeakdefinition, detector (whetheritisUVorMS)fastenough or lessinwidth.Itisimportanttoscanthe which maybeoftheorder1-2seconds Solvent Used(mL) Solvent Run Time (minutes) Time Run Resolution Backpressure 3.8 3.5 Gradient Time (min) Time Gradient Inj. volume (µL) volume Inj. Flow rate (mL/min) rate Flow Column: the gradient ontothecolumnin9.75 Atraditionalquaternarypumpwitha 100 x2.1mm100 µm, 2.6 RP-MS Accucore accuracy otherwise 2.4 6.0 2.5 218 35.0 35.0 60.0 500.0 35.0 %B 1.4 0.4 Solvent Used(mL) Solvent Run Time (minutes) Time Run Resolution Backpressure 3.0 1.9 1.7 Gradient Time (min) Time Gradient Inj. volume (µL) volume Inj. Flow rate (mL/min) rate Flow Column: 50 x2.1mm50 2.6µm, RP-MS Accucore 1.2 3.0 1.5 120 35.0 35.0 60.0 35.0 %B 0.7 0.4 Scaling Down a Method LC Columns and Accessories

Reasons to Scale Down a HPLC Typical Flow Rates for Analytical, Narrowbore, Capillary and Nanobore or LC/MS Method Columns (5µm Particles) There are applications where it is desirable to Column ID Flow Rate Range Optimum Flow Recommended API Source scale down a method without transferring the (mm) (µL/min) Rate1 (µL/min) Injection Volume2 (µL) method to U-HPLC. These reasons may be to: 4.6 1000 – 1500 1250 30 APCI or High  ow ESI • Maximize sensitivity when small amounts of 3.0 400 – 600 500 10 APCI or High  ow ESI sample are available 2.1 200 – 300 250 5 APCI or Micro-ESI 1.0 40 – 60 50 1 Micro-ESI • Make flow rate compatible with ionization technique in MS detection 0.5 10 – 25 12 0.35 Micro-ESI 0.32 4 – 10 5 0.15 Micro-ESI • Reduce costs by reducing solvent consumption 0.18 1 – 3 2 0.05 Micro-ESI Transfer Method to a Narrower Column 0.1 0.4 – 1 0.5 0.015 Nanospray Reducing the scale of a separation by reducing 0.075 0.2 – 0.5 0.3 0.01 Nanospray the column internal diameter may be 1. Recommended for good ef ciency and moderate pressure. Higher  ow rates may lead to column voids. Lower necessary when transferring a method from  ow rates are recommended for washing column bed or changing solvents.

UV to MS detection, or when only very small 2. Estimates based on negligible loss of ef ciency and isocratic elution with sample solvent identical to mobile >> phase. Larger volumes can be introduced under gradient conditions or using weaker sample solvent. amounts of sample are available, such as in Technical Information drug discovery or proteomics. In the first case Transfer Method to a Shorter Column Dwell Volume ionization technique or source design In gradient elution, the simplest way to reduce Dwell volume is just as important when scaling determines the best flow rate range (see table the method cycle time is to reduce the column down a method as for method transfer to above) and in the latter case, method length. If all other method parameters (column U-HPLC. The effect of dwell volume on the sensitivity is maximized because solutes elute ID and particle size, column chemistry, mobile separation is more significant when narrow in more concentrated chromatographic bands. phase composition, gradient range, flow rate, columns are used at low flow rates. For If all other method parameters (column length separation temperature) are kept unchanged instance, if the system has a dwell volume of and particle size, column chemistry, mobile the only requirement is to change the gradient 2.0mL and a 4.6mm ID column is run at 1mL/ phase composition, gradient range and time, time using the equation below, where gradient min, it takes 2 minutes for the gradient to separation temperature) are kept unchanged, time is reduced by the same factor as the reach the head of the column; however, if a the change to a narrower column only requires reduction in column volume. 2.1mm ID column is used with a 0.4mL/min adjustment of the flow rate. flow rate it will take 5 minutes for the tg1/Vc1= tg2/Vc2 F = F x (d /d )2 gradient to reach the column. In high 2 1 c2 c1 t – gradient time in original method (min) F – original flow rate (to be reduced) g1 throughput gradient separations using small 1 t – gradient time in new method (min) F – new flow rate g2 volume columns, dwell volume causes an 2 V – original column volume (mL) d – original column internal diameter c1 increase in run times and longer re- c1 V – new column volume (mL) d – new column internal diameter c2 equilibration time between runs. c2 Column volume V (mL) can be estimated using: This is applicable to both isocratic and c Several approaches can be taken to minimize V = 0.68 x π x r2 x L gradient methods. The new method should c these effects: produce a chromatogram with identical Vc – column volume (mL) • Select a pump with a small gradient delay volume resolution and identical run time. If small L – column length (cm) (e.g., Thermo Scienti c Accela high speed changes in retention times and resolution are r – column radius (cm) LC system has a delay volume of only 65µL); observed this is generally caused by system • Delay sample injection until gradient has dwell volume (discussed below). reached the head of the column; • Set the pump at a higher  ow rate and split the  ow before the column.

www.thermoscienti c.com/chromatography 4-233 4 -234 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories supply ofhighquality • • • Reasons toScaleUpanHPLCMethod Scaling Up aMethod column, ascalingfactorcanbeapplied. Once themethodisfinalizedonsmaller can alsobeusedtodeveloptheseparation. x 4.6mmpackedwiththemediaofinterest 250 chromatography. Scoutcolumns,typically and facilitatethescaleuptopreparative in varioussizestocomplementlabscaleoperations families ofThermoScientificphasesareoffered larger particlesizepreparativemedia.Theleading size mediaofferingthesameselectivityas analytical columnpackedwithsmallerparticle Scale upiseasiestwhenstartingfroman the ThermoScientificHypersilphases. our popularanalyticalsilicarangessuchas levels ofperformanceandreproducibilityas bonded phases,designedtogivethesame a strongreputation quickly oreconomically. We haveestablished dimensions, togivetheresultsyouneedmost and loadabilityofthemediaaswellcolumn preparative application,considertheselectivity best columnandpackingmaterialforyour compounds frommixtures.Inchoosingthe preparative sizestoisolateandpurify Analytical methodsmayrequirescaleupto Increasesamplethroughput Isolation andpurificationoftarget Increasemethodcapacity compounds for themanufactureand preparativesilicasand Mobile phaseviscositychangeswithcomposition calculated usingthisfactor. required forthenewlargerIDcolumnscanbe The columnloadingcapacityand ow rate d can alsobeusedtoestimatetheloading column packingmaterial.Thisscalingfactor internal diameterandtheparticlesizeof rate throughacolumnisdependentonits internal diameters.Thetypicalsolventflow constant betweencolumnswithdifferent peak retentiontimestoremainrelatively d capacity ofagivencolumn.Assumingcolumn of thecolumn.Thescalingflowratesallows required whenchangingtheinternaldiameter Flow rateandcolumnloadscalingareonly Scaling UptoaPreparativeColumn Scale Factor=d calculated usingthefollowingformula: length isaconstant,thescalefactorcanbe c2 c1 –newcolumninternaldiameter(mm) –originalcolumninternaldiameter(mm) c2 2 /d c1 2 and accuratedispensing Discover eVol XRforprecise PAGE

1-062 chromatography instrumentationbeingused. maximizing throughputwithrespecttothe variability isacriticalcomponentin addition ofmethanoloracetonitrile.This shows howwaterviscosityvarieswiththe composition. Asanexample,thefigureabove The mobilephaseviscosityvarieswith η =mobilephaseviscosity(centipoises) d =columndiameter(mm) d columnlength(mm) L = (1000for4.6mmIDcolumns) Φ =columnimpedance Pressure (atm)= following equation: cartridges canbecalculatedusingthe operating backpressureforcolumnsor may varywithsolventcomposition.Typical by theuseofgradients,wherepressure and solventflowrate.Itcanalsobeaffected particle size,temperature,solventproperties column length,internaldiameter, media Column operatingbackpressureisaffectedby Column Backpressure p =particlediameter(µm) 2.1 xΦL d p 2 xd 2 η Selecting the Media Peak Shape – Hypersil GOLD Media Peptides and Proteins – LC Columns and Accessories Media selection for your preparative In analytical HPLC, the use of packings based BioBasic and Hypersil GOLD Media separation is important. Choose media that on highly pure silica has been shown to When it comes to the analysis of peptides, the has a narrow particle size distribution which improve peak shape. Our highly developed and correct selection of packing material becomes will provide high efficiency columns with a reproducible manufacturing processes ensure ever more important. When deciding on which low back pressure, since there are no ‘fines’ that our leading analytical brand of Hypersil pore size of packing material to use in the to block frits or impede flow. The uniformly GOLD media is also available in a range of analysis of a polypeptide mix, molecular spherical particles, with narrow pore size particle sizes suitable for preparative LC weight and hydrophobicity of the peptides distribution, apparent in Thermo Scientific without compromise on performance. must be taken into account. Our breadth of preparative columns, provide reproducible silica offerings allow the chromatographer to Polar Compounds and Isomers – performance and a longer column life. Media obtain the best resolution using materials with Hypercarb Columns, Hypersil GOLD aQ that is available in a range of particle sizes pore diameters in the 100 to 300Å range. A Often when dealing with very polar compounds, offers choice for scale up applications with general rule is that hydrophilic peptides with achieving sufficient retention can be a controlled selectivity. We offers a range of a molecular weight of less than 2000 daltons challenge. We are able to offer a variety of choices for preparative media in several can be analyzed using a lower pore volume choices to overcome this common problem: particle sizes to tailor the media to your media, such as Hypersil GOLD media. Above The polar endcapping on Hypersil GOLD aQ application. this molecular weight, access to small pores

provides a controlled interaction mechanism is restricted, and separations tend to be

High Load and High Retention – by which moderately polar compounds can be >> inefficient. For hydrophobic peptides with a HyperPrep HS retained. Hypersil GOLD AX can be used in molecular weight greater than 2000, a 300Å Technical Information Materials with higher surface area can offer HILIC mode to provide retention of polar media such as Thermo Scientific BioBasic is increased loadability. This drive to maximize compounds. Hypercarb columns offer truly recommended. For the separation of small or surface area must be undertaken in a orthogonal selectivity to C18 in reversed hydrophilic peptides, a 100Å material such as considered manner particularly with regard to phase LC and can be used to retain highly HyperPrep HS may give better resolution. particle pore diameter and pore volume. Too polar compounds. Hypercarb columns can also high a pore volume will compromise stability be used to differentiate between very closely and robustness of the bed and too small a related compounds including geometric and pore diameter will influence mass transfer at positional isomers. the expense of efficiency. The high surface area provides enhanced retention of polar compounds. A high carbon loading gives a robust, stable phase. Please contact Technical Support for more information on Thermo Scientific HyperPrep columns and media.

www.thermoscienti c.com/chromatography 4-235 4 -236 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories for your quickreference. for unplugged beforestarting.Eyeprotectionshouldalsobeworn. Sheets (MSDSs)readilyavailable.Allelectricallypoweredinstrumentsshouldbeshutdownand chemical andphysicalpropertiesofanysolventsusedhavetheappropriateMaterialSafetyData Before youstartanytroubleshooting,itisessentialtoobservesafelaboratorypractices.Knowthe TroubleshootingHPLC The followingtablelistscommonHPLCproblemsencountered,thepossiblecausesandsolutions Symptom Pressure Low Pressure Related Problems Pressure High Baseline Fluctuating Problems Related Baseline Baseline Sloping Peaks Broad Peak Shape Problems Peaks Double Peaks Negative Cause Air in solvent lines or pump. Ensure that the reservoirs and solvent lines are fully primed and the purge valve is fully closed. fully is valve purge the and primed fully are lines solvent and reservoirs the that Ensure pump. or lines solvent in Air system. in Leak Low viscosity mobile phase. Con rm expected pressure using the Kozeny-Carmen or similar equation. similar or Kozeny-Carmen the using pressure Con expected rm leaking. seals Piston phase. mobile viscosity Low Sample precipitation. Consider sample clari cation steps such as as clari such sample SPE. or steps Consider cation ltration blockage. Detector precipitation. Sample blocked. Guard Tubing blocked. High viscosity mobile phase. Con rm expected pressure using the Kozeny-Carmen or similar equation. similar or Kozeny-Carmen the using pressure Con expected rm Pump malfunction. ow-rate phase. mobile viscosity High Old detector lamp. detector Old Contaminated solvents. contamination.Detector column. Contaminated guard. Contaminated  in Bubbles cell. ow System not equilibrated. Old detector lamp. detector Old frits. reservoir solvent Blocked properly. solvents mixing not Pump column. Contaminated Temperature uctuations. system. in Leak System not equilibrated. Gradient mobile phase. Contaminated solvents. Voided column. Voided column. Contaminated Old column. overload. mass avoid to concentration sample the Reduce cartridge. Old guard Temperature uctuations. high. too volume column expected the of Extra <40% of volumes injection Typically overload. avoid to volume injection the Reduce high. too mass Injected high. too volume Injection Injection solvent too strong. System not equilibrated. Voided column. Voided column. Contaminated Old guard cartridge. Old guard Ion pair method. pair Ion optics. detector RI Unbalanced Wrongly wired detector. Contaminated solvents. Action Check for and replace any leaking tubing or or tubing leaking any ttings. replace and for Check necessary. where replace and wear or leaking of evidence for Check Clean the  ow cell according to the manufacturer’s instructions.  manufacturer’s the the to Clean according cell ow Replace guard cartridge. necessary. as item replace and blockage of source check outlet, detector from backwards Working manufacturer.Contact Replace the lamp, particularly when this has been in use for > 2000 hours. >2000 for use in been has this when particularly lamp, the Replace grade. HPLC of solvents prepared freshly Use instructions.  manufacturer’s the the to Clean according cell ow column. the replace problem, the resolve not does this If solvent. appropriate an using column the Wash cartridge. guard the Replace limit. cell’s pressure the  the exceed not Do through ow-cell. solvent degassed pass and phase mobile the Degas phase. mobile of 10 volumes with column the Equilibrate Replace the lamp, particularly when this has been in use for > 2000 hours. >2000 for use in been has this when particularly lamp, the Replace methanol. then and water in frits reservoir the Ultrasonicate manually. solvents the blend isocratic, is method the If correctly. solvents the mixing is valve proportioning the that ensure used, being Where end. the at period awash has method agradient that Ensure solvent. appropriate an using column the Wash oven. column controlled athermostatically Use or tubing leaking any ttings. replace and for Check phase. mobile of 10 volumes with column the Equilibrate normal. is this Otherwise, wavelengths. higher or solvents purer Consider grade. HPLC of solvents prepared freshly Use Replace the column. Do not use outside the recommended pH range. pH recommended the outside use not Do column. the Replace column. the replace problem, the resolve not does this If solvent. appropriate an using column the Wash necessary. if column the Replace ef columns. new lower than much give ciencies columns Old methods. reverse-phase for reagents ion-pair with used been have that columns use not Do cartridge. guard the Replace peaks. sharper produce will temperatures Higher oven. column controlled athermostatically Use  the of volume possible. the where cell Reduce ow tubing. connecting of length and diameter Reduce used. be should width peak phase. mobile the than strength weaker or same the is solvent injection the that Ensure phase. mobile of 10 volumes with column the Equilibrate Replace the column. Do not use outside the recommended pH range. pH recommended the outside use not Do column. the Replace column. the replace problem, the resolve not does this If solvent. appropriate an using column the Wash cartridge. guard the Replace Inject the sample in the mobile phase. mobile the in sample the Inject instructions. manufacturer’s to Refer device. recording the to detector the from polarity signal the Check grade. HPLC of solvents prepared freshly Use LC Columns and Accessories Symptom Cause Action Peak Shape Problems Flat topped Detector overload. Reduce the sample concentration. Peaks Detector set-up. Check the detector attenuation and re-zero. Tailing Old guard cartridge. Replace the guard cartridge. Peaks Injection solvent too strong. Ensure that the injection solvent is the same or weaker strength than the mobile phase. Injection volume too high. Reduce the injection volume to avoid overload. Typically injection volumes of < 40% of the expected peak width should be used. Injected mass too high. Reduce the sample concentration to avoid mass overload. Old column. Do not use columns that have been used with ion-pair reagents for reversed phase methods. Old columns give much lower ef ciencies than new columns. Replace the column if necessary. Contaminated column. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Voided column. Replace the column. Do not use outside the recommended pH range. Fronting Old or damaged column. Replace the column. Peaks Peak Size and Retention Problems Small Degraded sample. Inject a fresh sample. Peaks Low analyte concentration. Increase the analyte concentration.

Detector set-up. Check the detector attenuation and re-zero.

No wash solvent. Check that the solvent wash reservoir is lled with a miscible solvent and that the injector is set to >> wash between injections. Damaged or blocked syringe. Replace the syringe. Technical Information Incorrect amount injected. Check injector loop size and that no more than 50% of this volume is used for partial loop injections. Viscous injection solvent. Reduce syringe draw-time. Old detector lamp. Replace the lamp, particularly when this has been in use for > 2000 hours. No Peaks Sample vial empty. Fill sample vial. Leak in system. Check for and replace any leaking tubing or ttings. Pump not mixing solvents Where being used, ensure that the proportioning valve is mixing the solvents correctly. If the method is properly. isocratic, blend the solvents manually. Damaged or blocked syringe. Replace the syringe. Different dwell volume. For gradient methods, check that the dwell volume of any new system is not very different from any previous system. Apply a nal hold time if necessary. Old detector lamp. Replace the lamp, particularly when this has been in use for > 2000 hours. Missing Degraded sample. Inject a fresh sample. Peaks Immiscible mobile phase. Check that any solvent already in the column is miscible with the mobile phase. Flush with propan-2-ol or ethanol where necessary. Fluctuations in pH. Buffer the mobile phase so that retention of ionizable compounds is controlled. Extra Peaks Degraded sample. Inject a fresh sample. Contaminated solvents. Use freshly prepared solvents of HPLC grade. Gradient methods often show ‘ghost-peaks’. Immiscible mobile phase. Check that any solvent already in the column is miscible with the mobile phase. Flush with propan-2-ol or ethanol where necessary. Fluctuations in pH. Buffer the mobile phase so that retention of ionizable compounds is controlled. Contaminated guard cartridge. Replace the guard cartridge. Contaminated column. Wash the column using an appropriate solvent. If this does not resolve the problem , replace the column. Varying System not equilibrated. Equilibrate the column with 10 volumes of mobile phase. Retention Leak in system. Check for and replace any leaking tubing or ttings. Temperature  uctuations. Use a thermostatically controlled column oven. Contaminated column. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Blocked solvent reservoir frits. Ultrasonicate the reservoir frits in water and then methanol. Pump not mixing solvents Where being used, ensure that the proportioning valve is mixing the solvents correctly. If the method is properly. isocratic, blend the solvents manually. Contaminated solvents. Use freshly prepared solvents of HPLC grade. Different dwell volume. For gradient methods, check that the dwell volume of any new system is not very different from any previous system. Apply a nal hold time if necessary. Piston seals leaking. Check for evidence of leaking or wear and replace where necessary. Air in solvent lines or pump. Ensure that the reservoirs and solvent lines are fully primed and that the purge valve is fully closed.

For more information, please request Successful HPLC Operation – A Troubleshooting Guide, TG20094.

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will helpdetermine unretained peak.Ameasurementofcapacity shown first: with theonesmostsensitivetopeakshape asymmetry. Themostcommonareshownhere, methods, someofwhicharesensitivetopeak d time changes). (capacity factor with retentiontimechanges)orthesystem Half-Height Tangential or 5-Sigma of peakbandspreadingdeterminedbyvarious Also numberoftheoreticalplates.Ameasure t Ef ciency(N) peak, wheret retention ofananalyterelativetounretained Expression thatmeasuresthedegreeof Capacity Factor(k) d flow rate(mL/min) = F =viscosity η column length(cm) = L Pressure Drop(psi)= mobile phaseviscosity(η),flowrate(F),column phase throughthecolumn.Itisrelatedto The pressurerequiredtopumpthemobile Backpressure De HPLC and Equations nitions 4-Sigma due tothecolumn sample peakandt length (L)anddiameter(d (d c p p = column internaldiameter(cm) = = particle diameter(µm) = ) bythefollowingequation: k = t R R W = tangential peakwidthor = W W = = W peak width at 50% W = = W peak widthat4.4% = 5.54(t N = 25(t N = 16(t N -t istheretentiontimefor 0 remains constantwithretention 0 (capacityfactorischanging 0 13.4% peakheight peak height peak height istheretentiontimeforan whether retention shifts are whether retentionshiftsare 250L R R /W) /W) d R p /W) 2 c d ), andparticlesize 2 2 η c

2 2 F

t concentration (apex). the tangential widthaverageofthepeaks: N isthenumberoftheoreticalplates. where Liscolumnlengthinmillimetersand 0.03mm. separation of theapextwopeaksdivided separation It canalsobeexpressedintermsofthe decreasing particlesize,changingtemperature, improved byincreasingcolumnlength, chromatographic peaks.Resolutioncanbe The abilityofacolumntoseparate Resolution (R where Liscolumnlengthandt velocity isgivenbythefollowingequation is directlyrelatedtocolumnpressure.Linear section. Thiseffectscolumnperformanceand The flowratenormalizedbythecolumncross Linear Velocity (or packed HPLCcolumnwith5µmparticles,HETP of acolumn’s efficiency. Foratypicalwell- carryover fromdistillationtheory:ameasure Height equivalenttoatheoreticalplate.A HETP where Fisflowrateinvolume/timeandt to eluteasolutefromthecolumnatmaximum Refers tothevolumeofmobilephaserequired Elution Volume (V the retentiontimeforpeakofinterest. 0.5(W changing breakthrough timeofanunretainedpeak: H ) valuesareusuallybetween0.01and theeluentorstationaryphase µ = R s L = R s 0 = s ) HETP =L/N (t V R R =F ) 2 –t 1 +W • t 1 R ) 2 ) 0 isthe . R by by is

in chromatography. Theequationrepresents The Btermisforthecontributioncausedby passing overparticlesofdifferentsizes. different pathsasolutemayfollowwhen describe eddydiffusion,whichallowsforthe and hasthreeterms.TheAtermisusedto van DeemterEquation or totallysymmetrical. Ideally, peaksshouldbeGaussianinshape from peakfronttoapexpointat5%height. peak widthat5%heightandfisthedistance by thefollowingequationwhereW A measureofthesymmetryapeak,given k Tailing Factor(T) composition. Wherek a certainstationaryphaseandmobile relative retentionoftwosubstances,fixedby A thermodynamicfactorthatisameasureof Selectivity (α) molecular diffusionorlongitudinalof the heightequivalentofatheoreticalplate(H) An equationusedtoexplainbandbroadening the solutewhilepassingthroughcolumn. mobile phaseasitpassesthroughthecolumn. The Ctermisthecontributionofmasstransfer respective capacityfactors. mobile phase.uisthelinearvelocityof solute betweenthestationaryphaseand and allowsforthefiniterateoftransfer H =A+ T =W α

= k 0.05 2 1 /2f B u + Cu 1 andk 2 arethe 0.05 isthe

Selecting the Right Buffer LC Columns and Accessories

A partial list of common buffers and their Common Buffer Systems corresponding pH values is shown in the Common Buffer pK Useful pH Range MS-Compatible Buffer Systems table. Perhaps the most common a TFA 0.30 Yes HPLC buffer is some form of phosphoric acid. Phosphate pK 2.1 1.1 – 3.1 No Remember that a true buffer should have the 1 pK 7.2 6.2 – 8.2 No ability to resist pH change when a sample is 2 pK 12.3 11.3 – 13.3 No introduced at a different pH, and that buffer capacity 3 is only 100% at the pK value of the acid or base. Citrate pK1 3.1 2.1 – 4.1 No At pH 4, phosphate is a poor buffer and would pK2 4.7 3.7 – 5.7 No

pK3 5.4 4.4 – 6.4 No change rapidly toward one of its pKa values if a more acidic or basic sample were introduced. Formate 3.8 2.8 – 4.8 Yes As a rule, one should work within ±1pH unit of Acetate 4.8 3.8 – 5.8 Yes Tris Base (Trizma, THAM) 8.3 7.3 – 9.3 Yes the buffer pKa value for good pH control of the mobile phase. Adequate buffer concentrations Ammonia 9.2 8.2 – 10.2 Yes

for HPLC tend to be in the 10-100 millimolar Borate 9.2 8.2 – 10.2 No level depending on the size and nature of the Diethylamine 10.5 9.5 – 11.5 Yes >> Carbonate pK 6.4 5.4 – 7.4 Yes sample, as well as the column packing material. 1 Technical Information

Phases based on highly pure silica with robust pK2 10.3 9.3 – 11.3 Yes bondings such as the Hypersil GOLD range, are Triethanolamine 7.80 Yes often more compatible with dilute buffers than traditional packings. When control at a lower pH (2-3) is desired, 3 2 A: pH 2.1 B: pH 2.5 BETASIL C18, 5µm, 50 x 4.6mm 2 3 phosphate, or stronger organic acids such as Part Number: 70105-054630 1 7 Eluent: 50% ACN / 50% TFA or acetic acid, are commonly used. If control 1 7 25mM KH3PO4 at pH 4-5 is desired, an organic acid buffer at pH indicated Flow Rate: 0.8mL/min such as acetate or citrate should be considered Detector: UV @ 220nm in place of phosphate. Sample: 1. Uracil 2. Tolmetin The figure to the right shows the importance of 3. Naproxin 4 5 4. Fenoprofen choosing the correct pH for a separation. Even 4 5 . D i  u n i s a l 5 6. Indometacin slight changes in pH, either from measuring errors, 7. Ibruprofen mixing complications with the pump, or atmospheric 6 6 water adsorption into the mobile phase, can alter any method if not properly buffered. 701-085 701-086 Care should be taken when choosing a buffer and organic modifier mixture to ensure that a solution of the two does not produce a solid salt which 0 6 MIN 0 6 MIN could cause blockages and system contamination. Effect of small changes in pH on the separation of mildly ionizable compounds Buffers should always be flushed from the analytical column and instrument after use to avoid salts being deposited on delicate frits etc.

Buffer Selection for LC/MS

Buffer choice will be very dependent on the • Ammonium acetate/formate/hydrogen Note: There are LC/MS instruments available, for analyte and the instrumentation used. Ideally, carbonate (< 50mM) example the Thermo Scienti c Surveyor MSQ LC/MS, which incorporate a self-cleaning mechanism to LC/MS applications should use a volatile • Formic/acetic acid (0.01 – 1% v/v) reduce the build up of inorganic buffers on the source during routine use. Care should still be taken not to buffer as this will not form a contaminating • Tri uoroacetic acid (< 0.1% v/v) deposit on the source. Inorganic acids, involatile purposefully over-contaminate the instrument source • Trialkylamine (< 0.1% v/v) and aqueous as this will lead to operating dif culties. buffers and ion-pair reagents should all be ammonia type bases avoided. Typical LC/MS buffers include: • TRIS • BIS-TRIS propane

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Response (Millivolts) -20 20 40 0 0 Baseline noisefromgasinmobilephase store HPLCgradewateringlasscontainers. leach intothewaterandcontaminateit.Always plastic containers.Additivesinthemay Important: DonotstoreHPLCgradewaterin purchased fromsolventsuppliers. volumes. Preferably, HPLCgradewatercanbe to producewaterofsuitablequalityinhigh purification instrumentsusethismechanism through anionexchangebed.Modernwater is generatedbypassingdeionizedwater use. UltrapureHPLCwater(18mΩresistivity) and thereforeisnotrecommendedforHPLC contains tracelevelsoforganiccompounds is ofthehighestpurity. Deionizedwateroften Ensure thatanywaterusedinbufferpreparation spurious baselinepeaks. trace levelsofimpurities,whichmayproduce whereas lowergradereagentsdocontain spurious peaksinachromatogrambaseline reagents containnoimpuritiestoproduce the differenceinpurityismarked.HPLCgrade slightly morethanlowergradereagents,but highest quality. HPLCgradereagentsmaycost All reagentsandsolventsshouldbeofthe Quality troubleshooting spuriouspeaks,baselinenoiseetc. Correct solventpreparationisveryimportant.Itcansavevastamounts oftimespent Mobile Phases of Preparation Mobile Phase–NotDegassed 2 4 Minutes 6 so itisoftenworthbuying chromatographic behaviorofbuffersolutions, stabilizing agentsoftenaffecttheopticaland for example,sodiummetabisulphite.These Buffer reagentscancontainastabilizingagent, buffer shelflife. monographs orsimilarforfurtherguidanceon they haveafinitelifetime.Refertopharmacopoeia If buffersolutionsarestored,beawarethat affect chromatography. Changes inpHandmicrobialgrowthwill easily contaminatedby prevent re-contaminationwithdustetc. should bestoredinacoveredreservoirto cause blockages.Afterfiltration,thesolvents removes anyparticulatematterthatmay through a0.45µmfilterbeforeuseThis Ideally, allHPLCsolventsshouldbefiltered Filtration and thatthereisnomicrobialgrowthpresent. buffer pHisunaffectedbyprolongedstorage day required.Thispracticeensuresthatthe All buffersshouldbepreparedfreshlyonthe Buffers no stabilizer. Containers reason, werecommend purchased inlowcontainerweights. 8

Response (Millivolts) -20 20 40 0 0 ofsolidreagentare that reagentsbe repeated use.Forthis reagents that contain reagents thatcontain Mobile Phase–Degassed 2

4 Minutes of pressureinsidethereservoir. vent totheatmospherepreventbuildup Note: Ensurethatthesolventreservoirhasa re-adsorption ofgasesovertheanalysistime. throughout theanalysis.Thiswillinhibit with heliumoranotherlowsolubility • • • solution by: gasses. Dissolvedgascanberemovedfrom thoroughly degassedtoremovealldissolved pumped aroundtheHPLCsystem,itshouldbe Before thefreshlypreparedmobilephaseis Degassing The mostefficientformofdegassingis spurious baselinepeaksetc. method isavailable,werecommend can forminthelowpressureof If themobilephaseisnotdegassed,airbubbles cell resultinginproblemswithsystem phase iscontinuallydegassed maximized. valves willperformbetterandlifetimesbe HPLC system.Pumpplungers,sealsandcheck chromatography andtheweartearof Filtering HPLCsolventswillbenefitbothyour Vacuum ltration Sonication Bubblingwithhelium 6 8 at very low levels at verylowlevels

that the mobile that themobile instability, detector

gas. Ifthis bubbling bubbling Solvent Properties (vs Silica Gel) and Miscibility LC Columns and Accessories

>> Technical Information

www.thermoscienti c.com/chromatography 4-241 4 -242 Thermo Scienti c Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories exhaustive list. information containedinthefollowingtableis have oneormoredetectionwavelengths,eachofwhichhasamolaradsorbtivity associatedwithit.The Chromophores arelightabsorbinggroups. Wavelengths Detection Chromophore Nitro Nitrile Thiol Thioether Sulfoxide Sulfone Pyridine Oxime Nitroso Nitrate Napthalene Ketone Ethylene Ether Ester Carboxyl Benzene Azo Azidin Amine Aldehyde Acetylide Chromophore -C==C- -NO -C==N -ONO -SH -S- > S-O 215 215 -SO -NOH -N=O -ONO > C=O -C=C- -O- -COOR -COOH -N=N- > C=N- C=C-NO C=C-C=O C=C-C=N C=C-C=C -(C=C)- -(C=C) -NH -CHO N 2 2 2 - 2 3 2 acyclic 2

Their behaviorisusedtoallowthedetection intended asaguidetocommonchromophores. 175 –180 λ 300 –400 300 160 220 –230 220 –350 300 195 194 210 1,600 180 251 195 174 190 302 210 260 270 312 275 220 –285 270 195 190 185 205 –210 200 255 202 184 –400 285 190 195 210 –250 220 219 210 –230 280 –300 280 210 max (nm) max 6,000 ε 10 1,000 – 2,000 – Weak 1,400 4,600 1,500 strong Very 1,700 6,000 80,000 5,000 100 Strong 10,000 – 20,000 23,000 6,500 35,000 12 5,600 175 112,000 18 –30 1,000 8,000 1,000 50 –70 50 170 6,900 46,700 3 –25 5,000 2,800 21,000 11 –18 Strong max (L/m/cm) max ofanalytes.They Itisnotan Column Cleaning and Regeneration LC Columns and Accessories

Testing of column performance can be undertaken Protein Size Exclusion Media Normal Phase Regeneration using the experimental conditions in the test There are two wash/regeneration procedures Suitable for applications running predominantly certificate provided with the column. The column associated with the removal of contaminants in normal phase mobile phases. efficiency, capacity factor, etc. should be measured from protein size exclusion media. 1. Flush with 50mL dichloromethane at the start and end of the clean-up procedure Weakly Retained Proteins 2. Flush with 50mL methanol to ensure that it has been performed successfully 1. Flush with 30mL 0.1M pH 3.0 phosphate buffer 3. Flush with 50mL water and has improved the column performance. 4. Flush with 50mL 0.1M hydrochloric acid In all instances, the volume of solvent used is Strongly Retained Proteins 40 – 60 column volumes unless otherwise stated. 1. Flush for 60 minutes using a 100% water 5. Flush with 50mL water Ensure that no buffers or samples are present to 100% acetonitrile gradient 6. Flush with 50mL methanol on the column and that the solvent used prior Porous Graphitic Carbon 7. Flush with 50mL dichloromethane to the clean up is miscible with the first wash There are four wash or regeneration procedures 8. Flush with mobile phase until equilibrated solvent. After the clean up, ensure that the test associated with porous graphitic carbon. The Author: A. Karlsson – Uppsala, Sweden mobile phase is miscible with the last solvent one(s) used will depend on the analytes and Removal of TFA and DEA in the column. solvents that have been used with the column TFA and DEA have the potential to adsorb to Normal Phase Media Acid/Base Regeneration the surface of porous graphitic carbon; after >> 1. Flush with tetrahydrofuran Suitable for ionized species analyzed in strongly using these additives in the mobile phase, Technical Information regeneration of the column should be undertaken 2. Flush with methanol aqueous mobile phases. to ensure the original Hypercarb selectivity 3. Flush with tetrahydrofuran 1. Invert the column and optimum performance will always be 2. Flush with 50mL tetrahydrofuran:water (1:1) 4. Flush with methylene chloride achieved. The regeneration is as follows: containing 0.1% tri uoroacetic acid 5. Flush with benzene-free n-hexane 1. Removal of TFA: Flush column with 70 column 3. Flush with 50mL tetrahydrofuran:water (1:1) Reversed Phase Media volumes of THF. containing 0.1% triethylamine or sodium 1. Flush with HPLC grade water; inject 4 hydroxide 2. Removal of DEA: Set column oven to 75°C aliquots of 200µL DMSO during this  ush and  ush column with 120 column volumes 4. Flush with 50mL tetrahydrofuran:water (1:1) of ACN. 2. Flush with methanol containing 0.1% trifluoroacetic acid 3. Flush with chloroform 5. Flush column with 70 column volumes of THF Polymeric Media with Metallic Counter Ions 4. Flush with methanol 6. Flush with methanol/water (95:5) There are three types of regeneration available Anion Exchange Media to re-equilibrate for polymeric columns with metal counter ion. 1. Flush with HPLC grade water 7. Re-invert the column Details of each procedure are listed in the Author: R. Plumb – Glaxo, UK 2. Flush with gradient of 50mM to 1M following table. appropriate buffer solution Strong Organic Regeneration 3. Flush with HPLC grade water Suitable for applications involving polar and/ or 4. Flush with methanol ionized species analyzed in aqueous mobile phases. 5. Flush with chloroform 1. Flush with 50mL acetone 2. Flush with 120mL dibutylether Cation Exchange Media 3. Flush with 50mL acetone 1. Flush with HPLC grade water; inject 4 aliquots of 200µL DMSO during this  ush 4. Flush with aqueous mobile phase until equilibrated 2. Flush with tetrahydrofuran

Column Type Metal Contamination Organic Contamination Column Cleaning Hydrogen Counter Ion Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min

with 0.1M H2SO4 @ 25°C for 4 to 16 hr with 20:80 ACN: H2O @ 25°C for 4 hr with 20:80 ACN: 0.01M H2SO4 @ 65°C for 4 hr Calcium Counter Ion Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min

with 0.1M Ca(NO3)2 @ pH 6.3 and 85°C with 20:80 ACN:H2O @ 25°C for 4 hr with 20:80 ACN:H2O @ 25°C for 4 hr for 4 to 16 hr Sodium Counter Ion Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min

with 0.1M NaNO3 @ 85°C for 4 to 16 hr with 20:80 ACN:H2O @ 25°C for 4 hr with 20:80 ACN:H2O @ 25°C for 4 hr Lead Counter Ion Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min Pump in reverse  ow mode at 0.1mL/min

with 0.1M Pb(NO3)2 @ pH 5.3 and 85°C with 20:80 ACN: H2O @ 25°C for 4 hr with 20:80 ACN: H2O @ 25°C for 4 hr for 4 to 16 hr

www.thermoscienti c.com/chromatography 4-243 Thermo Scientific Resources

for Chromatographers Chromatography Columns and Consumables

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