T h e r m o S c i e n t i f i c C h r o m a t o g r a p h y C o Thermo Scientific lu m Chromatography Columns n s and Consumables 2014-2015 a n d C o n s u m a b le s 2 0 1 4 - 2 0 1 5 Your world of chromatography predictability LC Columns and Accessories As a leader in LC column technology including silica, polymer and porous graphitic carbon manufacturing, bonded phase production and column packing for over 5 1 0 4-2 35 years, you can rely on the quality of Thermo Scientific HPLC products. Here we 1 0 2 s showcase our latest and most comprehensive range of innovative columns, accessories e bl a m and equipment for fast LC, analytical HPLC and biomolecule separations. u s n o C d n a s n m u ol C y h p a gr o at m o hr C c fi nti e ci S o m er h T 4 L C Section Contents C o l u Introduction . . . . . . . . . . . . . . . . . . 4-001 LC Accessories. . . . . . . . . . . . . . . . 4-158 m Column Selection. . . . . . . . . . . . . . 4-002 LC Equipment. . . . . . . . . . . . . . . . . 4-186 n Column Protection . . . . . . . . . . . . . 4-016 LC Reagents. . . . . . . . . . . . . . . . . . 4-193 s a Columns for Fast and Analytical LC . . 4-022 LC and LC-MS Instrument Parts . . 4-209 n Columns for Biomolecules. . . . . . . 4-110 Technical Information. . . . . . . . . . . 4-231 d A c c e s s o r i e Featured Products s 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 effi 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.thermoscientifi c.com/chromatography 4-001 s LC Column Selection e i r o Information in the following section will help you make an informed decision on the s s appropriate HPLC column for your application, based on stationary phase use, analyte e c properties, LC-MS requirements or USP specifications. You will also find a useful table c of Thermo Scientific phases with specifications, as well as recommended Thermo A Scientific alternatives for other popular columns. d n a Refer to the Advanced User Graphic (AUG) on the The Pore Size is shown, with larger pore size columns Hydrophobicity s corresponding product page (illustrated to the right) being more applicable to larger analytes such as n Low High m for more help and information on general purpose proteins or peptides. The percentage Carbon Load pH Range u column selection. is related to the hydrophobicity. Below the icon, you ol The AUG will show you Hydrophobicity which gives will see the particle sizes available, as well as the P0ore Size 2 to 8 14 C USP code. These graphics are designed to allow you the relative retention on the column. Generally, the C higher the hydrophobicity, the greater the retention to quickly compare the main characteristics of multiple 0 80Å 300 Carbon Load (%) L of neutral compounds and the higher the organic stationary phases, allowing you to choose quickly the most appropriate stationary phase for your analysis. 0 5 25 content in the mobile phase. A lower value indicates Particle Size2.6µm USP L11 a need for higher aqueous mobile phases to achieve For additional help in column selection, please comparable retention and resolution. The see the back cover to contact our expert Technical Advanced User Graphic (AUG) recommended pH Range for the column is illustrated, Support and tap into our expertise to help make outside of which column lifetimes will diminish. the best choice for your application. Common HPLC Phases and Their Uses Common Alternative Functional Normal Reverse Ion HILIC Application Name Name Group Phase Phase Exchange Silica Silica -OH (cid:127) (cid:127) Non-polar and moderately polar organic compounds. C1 SAS -(CH) Least retentive of all alkyl group bonded phases for non-polar solutes. 33 (cid:127) Typically used for moderately polar and multi-functional compounds. C4 Butyl -CH (cid:127) Shorter retention than C8, C18. Separation of peptides and proteins. 4 9 C8 MOS -CH Less retentive than C18; normally used for small peptides and 8 17 (cid:127) proteins, pharmaceuticals, steroids, environmental samples. C18 ODS -C H Most retentive of the alkyl-bonded phases. Used widely for 18 37 (cid:127) pharmaceuticals, steroids, fatty acids, phthalates, environmental etc. Cyano CPS, CN -(CH)CN Unique selectivity for polar compounds, more suitable than base 23 silica for normal phase gradient separations. When used in reversed (cid:127) (cid:127) phase, the selectivity is different to that of the C8 and C18 phases. Useful for a wide range of pharmaceutical applications and for 5 01 mixtures of very different solutes. 2 14- Amino APS -(CH)NH HILIC: Carbohydrate analysis and other polar compounds. s 20 23 2 (cid:127) (cid:127) (cid:127) (cid:127) Weak anion exchange: anions and organic acids. ble Normal Phase: Alternative selectivity to silica. Good for aromatics. a m Phenyl -(CH)CH (cid:127) Aromatic compounds and moderately polar compounds. u 3 6 5 s n Pentafl uo- PFP -CF Extra selectivity and retention for halogenated, polar compounds d Co rophenyl 6 5 (cid:127) and isomers. n s a Diol -(CH2)2O (cid:127) (cid:127) (cid:127) Reversed Phase: Proteins, peptides. mn CH2(CH2OH)2 Normal Phase: Similar selectivity to silica, but less polar. u Col SCX Strong Cation -RSO3H- (cid:127) Organic bases. y Exchanger h p a SAX Strong Anion -RN+(CH) Organic acids, nucleotides and nucleosides. ogr Exchanger 33 (cid:127) at m AX Anion Exchanger -(CHCHNH-) Organic acids, nucleotides and oligonucleotides. hro Polyethyleneimine 2 2 n (cid:127) C c (PEI) fi nti Porous PGC 100% carbon Particularly useful for the separation of highly polar compounds that e ci graphitic are diffi cult to retain using conventional silica based columns; S (cid:127) (cid:127) o carbon separation of structurally similar compounds (e.g., isomers, m er diastereoisomers). h T 4-002 HPLC Stationary Phase Column Selection L C Before beginning a new analysis, consider the physical and chemical properties of the analyte(s), the mode of analysis and how the C analyte(s) will interact with the surface of the chromatographic phase. To aid column selection, the following guide may be useful. o l u m Non-polar n s Analyte Analyte Chemical Properties Mode of Separation Recommended Phase a Solubilty n d Acclaim Mixed-Mode WAX-1 4-069 Acidic pKa > 2 Anion exchange / Reversed-phase mixed mode Acclaim Trinity P1 4-091 A c Accucore C18 4-045 c Accucore RP-MS 4-044 e s Accucore C8 4-046 s Accucore C30 4-052 o r Accucore Phenyl-Hexyl 4-049 i e Acclaim 120 C18 4-061 s Non-polar Reversed phase Acclaim 120 C8 4-062 > Acclaim C30 4-064 > AHcycplearismil PGhOeLnDy l-1 44--002663 C o Hypersil GOLD C8 4-027 lu m Hypersil GOLD C4 4-028 n Neutral Syncronis C18 4-075 S e Syncronis C8 4-076 le c Accucore aQ 4-047 tio Accucore Polar Premium 4-048 n Accucore PFP 4-051 Accucore Phenyl-X 4-050 Acclaim PolarAdvantage 4-065 Polar Moderately-polar Reversed phase Acclaim PolarAdvantage II 4-066 solvents 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.thermoscientifi c.com/chromatography 4-003 s HPLC Stationary Phase Column Selection continued e i r o Polar s s e Analyte c Analyte Chemical Properties Mode of Separation Recommended Phase c Solubilty A Polar retention effect on graphite Hypercarb 4-084 d Hypersil GOLD AX 4-034 n Anion exchange a Hypersil GOLD SAX 4-035 Acidic pK < 2 Accucore Urea-HILIC 4-054 s a n Accucore 150-Amide-HILIC 4-146 m HILIC Acclaim HILIC-10 4-067 u Syncronis HILIC 4-081 l o Polar retention effect on graphite Hypercarb 4-084 C Accucore HILIC 4-053 C Acclaim Mixed-Mode HILIC-1 4-068 L Acclaim Mixed-Mode WAX-1 4-069 Polar Neutral Acclaim Mixed-Mode WCX-1 4-070 solvents HILIC Acclaim Trinity P1 4-091 Hypersil GOLD Amino 4-033 Hypersil GOLD HILIC 4-037 Syncronis Amino 4-079 Syncronis HILIC 4-081 Polar retention effect on graphite Hypercarb 4-084 Acclaim Mixed-Mode WCX-1 4-070 Cation exchange Acclaim Trinity P1 4-091 Basic pK > 10 Accucore Urea-HILIC 4-054 a Accucore 150-Amide-HILIC 4-146 HILIC Acclaim HILIC-10 4-067 Syncronis HILIC 4-081 Hypersil GOLD Amino 4-033 Hypersil GOLD CN 4-032 Non-polar Normal phase Hypersil GOLD Silica 4-036 solvents Syncronis Amino 4-079 Syncronis Silica 4-080 The above table recommends columns for the separation of small molecule analytes. Column recommendations for the separation of biomolecules can be found in the table on page 4-111 5 1 0 2 4- 1 0 2 s e bl a m u s n o C d n a s n m u ol C y h p a gr o at m o hr C c fi nti e ci S o m er h T 4-004 Column Format Selection for LC-MS L C The Thermo Scientific range offers a broad array of column designs and stationary phases optimized for C LC-MS applications. Use the following table to help you choose your column format to best meet your o application needs. A variety of HPLC column hardware configurations are available, designed to give superior lu results for high speed, high sensitivity, high efficiency and convenience. A wide range of particle and m monolithic stationary phases allows choices for optimized selectivity. n s Column Hardware Selection for LC-MS a n LC-MS Application Column Hardware Design Description d High throughput analysis Javelin HTS columns Direct-connection columns A Slim design, 20mm length, 1mm to 4.6mm ID c High sensitivity analysis Acclaim PepMap nano, capillary Nano, capillary and micro columns 0.075mm to 1mm ID; 50 to 250mm length c and micro columns nanoViper format offers fi ngertight dead volume free connection to 1000 bar e s Proteomics analysis EASY-Spray columns Combined column/emitter design with nanoViper connection. s Heated fl exible silica columns 50µm and 75µm ID; 150 to 500mm length o Acclaim PepMap nano columns, nanoViper offers fi ngertight dead volume free connection to 1000 bar ri e Accucore nano columns, nanoViper Flexible silica columns 50μm and 75μm ID; 50 to 500mm length s 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 > EASY-Column FTrleaxpi bcloel usmilinc a2 0comlumm xn 1s0 100μ0mm mID x 75μm C o PepSwift monolithic columns Flexible fused silica columns, nanoViper connections lu m 100µm to 500µm ID, 50 to 250mm length n Trap columns 5mm x 200µm ID S e Various HPLC columns, throughout this LC section, can also be used for LC-MS application. Typical flow rates and MS source le c t compatibility for these columns are shown in the table on the next page io n wwwwww..tthheerrmmoosscciieennttiififi cc..ccoomm//cchhrroommaattooggrraapphhyy 4-005 s Column Selection for LC-MS continued e i r o Packed column selection for LC-MS s s e Analyte c Molecular Sample Interface Relative Column ID Flow Rate c Weight Polarity Ionization Sensitivity (mm) (µL/min) Column Hardware A Low 4.6, 4.0, 3.0 2000 – 200 d Javelin HTS, Analytical n Low APCI a High 2.1, 1.0 200 – 50 Analytical, Javelin HTS s n Low 4.6, 4.0, 3.0 2000 – 200 Javelin HTS, Analytical m APCI High 2.1, 1.0 200 – 50 u Analytical, Javelin HTS l < 1000 Da Medium o Low 2.1, 1.0 200 – 50 C Analytical, Javelin HTS ESI 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro C High 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano L Low 2.1, 1.0 200 – 50 High Analytical, Javelin HTS (or ionizable) ESI 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro High 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano Low 2.1, 1.0 200 – 50 Analytical, Javelin HTS > 1000 Da ESI 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro High 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano Monolith columns for LC-MS Analyte Molecular Weight Column ID (mm) Flow Rate (µL/min) Column Hardware < 1000 Da 0.1, 0.2, 0.5 0.7 – 25 PepSwift Monolith > 1000 Da 1.0 40 – 200 ProSwift Monolith ProSwift is also available in larger IDs for high throughput applications. 5 1 0 2 4- 1 0 2 s e bl a m u s n o C d n a s n m u ol C y h p a gr o at m o hr C c fi nti e ci S o m er h T 4-006 HPLC Column Selection by U.S. Pharmacopeia Specifi cations* L C C USP Code Description Recommended Phase Page o L1 Octadecyl silane (C18) chemically bonded to porous Acclaim 120 C18 4-061 l u or ceramic microparticles, 1.5 to 10μm in diameter, Acclaim 300 C18 4-142 m or a monolithic rod Accucore C18 4-045 n Accucore aQ 4-047 s Accucore 150-C18 4-144 a n Accucore XL C18 4-056 d BioBasic 18 4-140 A Hypersil GOLD 4-026 c Hypersil GOLD aQ 4-029 c e Acclaim PepMap 100 C18 4-149 s Syncronis C18 4-075 s o Syncronis aQ 4-077 r L3 Porous silica particles, 1.5 to 10μm in diameter, Accucore HILIC 4-053 ie or a monolithic rod Hypersil GOLD Silica 4-036 s Syncronis Silica 4-080 > L7 Octyl silane chemically bonded to totally Acclaim 120 C8 4-061 > o1.r5 s tuop 1e0rfiμ cmia ilnly d piaomroeutse sr,i loicra a p maortnicolleitsh, ic rod Accucore C8 4-046 C Accucore XL C8 4-057 o lu BioBasic 8 4-140 m n Hypersil GOLD C8 4-027 S Acclaim PepMap 100 C8 4-150 e le Syncronis C8 4-076 c t L8 An essentially monomolecular layer of Hypersil GOLD Amino 4-033 io n 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.thermoscientifi c.com/chromatography 4-007 s HPLC Column Selection by U.S. Pharmacopeia Specifi cations* continued e i r o s s USP Code Description Recommended Phase Page e c L17 Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate H+ 4-156 c cross-linked styrene-divinylbenzene copolymer in the HyperREZ XP Organic Acids 4-156 A hydrogen form, 6 to 12μm in diameter d L19 Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate 4-156 n cross-linked styrene-divinylbenzene copolymer in the Ca2+ a calcium form, about 9μm in diameter HyperREZ XP Sugar Alcohols 4-156 s L20 Dihydroxypropane groups chemically bonded to BETASIL Diol Inquire n porous silica or hybrid particles, 1.5 to 10μm m in diameter u L26 Butyl silane chemically bonded to totally porous silica Accucore 150-C4 4-145 ol particles, 1.5 to 10μm in diameter BioBasic 4 4-140 C Hypersil GOLD C4 4-028 C Acclaim PepMap 300 C4 4-149 L L33 Packing having the capacity to separate dextrans BioBasic SEC 120 4-139 by molecular size over a range of 4,000 to 500,000 BioBasic SEC 300 4-139 daltons. It is spherical, silica-based, and processed to provide pH stability BioBasic SEC 1000 4-139 MAbPac SEC-1 4-129 L34 Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate Pb2+ 4-156 cross-linked styrene-divinylbenzene copolymer in the lead form, 7 to 9μm in diameter L43 Pentafl uorophenyl groups chemically bonded to silica Accucore PFP 4-051 particles by a propyl spacer, 1.5 to 10μm in diameter Hypersil GOLD PFP 4-030 L46 Polystyrene/divinylbenzene substrate agglomerated OmniPac PAX-100 4-103 with quaternary amine functionalized latex beads about 9 to 11um in diameter L50 Multifunction resin with reverse-phase retention and OmniPac PAX-500 4-103 strong anion-exchange functionalities. The resin consists of ethylvinylbenzene, 55% cross-linked with divinylbenzene copolymer, 3 to 15µm in diameter, and a surface area of not less than 350m2 per g. Substrate is coated with quaternary ammonium functionalized latex particles consisting of styrene cross-linked with divinylbenzene L52 A strong cation exchange resin made of porous silica BioBasic SCX 4-141 with sulfopropyl groups 5 to 10μm in diameter L58 Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate Na+ 4-156 cross-linked styrene-divinylbenzene copolymer in the sodium form, about 6 to 30μm in diameter 5 01 L59 Packing for the size exclusion separation of proteins BioBasic SEC 300 (5μm) 4-139 14-2 (separation by molecular weight) over the range of MAbPac SEC-1 4-129 0 5 to 7,000 kDa. The packing is spherical, 1.5 to 10um, 2 s silica or hybrid packing with a hydrophilic coating e bl a L60 Spherical, porous silica gel, 10μm or less in diameter, Acclaim PolarAdvantage 4-065 m u the surface of which has been covalently modifi ed s Acclaim PolarAdvantage II 4-066 on with alkyl amide groups and endcapped d C Accucore Polar Premium 4-048 n s a L62 C30 silane bonded phase on a fully porous spherical Acclaim C30 4-064 mn silica, 3 to 15µm in diameter Accucore C30 4-052 u Col L78 A silane ligand that consists of both reversed-phase Acclaim Mixed-Mode WAX-1 4-069 hy (an alkyl chain longer than C8) and anion-exchange p gra (primary, secondary, tertiary or quaternary amino ato groups) functional groups chemically bonded to m porous or non-porous or ceramic micro-particles, o hr 1.0 to 50μm in diameter or a monolithic rod C c fi * These are the recommended Thermo Scientific HPLC columns for various USP categories although other columns enti for each category are also available. ci S o m er h T 4-008