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HICHROM Chromatography Columns and Supplies LC COLUMNS Catalogue 9 Hichrom Limited The Markham Centre, Station Road Theale, Reading, Berks, RG7 PE, UK Tel: + (0)8 90 660 Fax: + (0)8 9 8 Email: sales@hichrom.co.uk www.hichrom.co.uk

Please contact Hichrom for current prices. Tel: + (0)8 90 660 Email: sales@hichrom.co.uk SHODEX Silica- and polymer-based phases Columns for all chromatographic modes Wide application range Capillary, microbore, analytical and preparative dimensions Showa Denko K.K., Japan manufactures the line of HPLC columns for the pharmaceutical and biotechnology industries. A wide range of products with both silica-based and polymer-based materials is available. An overview of the products is shown on the following pages: reversed-phase (page 0), ligand exchange (page ), ion-exchange and ion chromatography (page ), size exclusion (page ), HIC (page ) and affinity (page ). Please enquire for additional details. Reversed-Phase Columns The product range contains both polymer-based and silica-based phases. Please enquire for ordering details where not listed. Reversed-Phase (Polymer-based) Phases The table below lists the polymer-based phases available. RSpak RP8, DS Styrene-divinylbenzene - Analysis of proteins and peptides RSpak DE, GOLF, CARB Polymethacrylate - DE series columns have similar selectivity to C8 columns RSpak DM-6 Polyhydroxymethacrylate - Suitable for analysis of amino acids and polypeptides RSpak NN Polyhydroxymethacrylate Sulpho Suitable for separation of mixtures of neutral and ionic RSpak JJ-50 Polyvinyl alcohol Quaternary ammonium compounds Asahipak ODP Polyvinyl alcohol Octadecyl Asahipak ODP HP Polyhydroxymethacrylate Octadecyl Large pore size phases, suitable for high molecular weight compounds eg. proteins, as well as low molecular weight Asahipak C8P-50 Polyvinyl alcohol Octyl compounds. LC-MS applications Asahipak CP-50 Polyvinyl alcohol Butyl Asahipak NHP Polyvinyl alcohol Amino Analysis of saccharides (NP) ODP HP is a macroporous polyhydroxymethacrylate reversed-phase material offering better efficiencies than many other resin-based phases and comparable to that of silica-based C8 columns. It has a recommended ph range of to. The high polarity and small pore size (0Å) of this phase is designed to exclude proteins and can be used for the analysis of drugs in biological samples ie. as restricted access material (see Figure for example). Blank switch valve introduce to MS UV, 0nm with BSA BSA switch valve introduce to MS UV, 0nm Sample: 5μl : 500μg/l BSA: 7.0mg/ml 0.0 0.5.0.5.0.5.0.5.0.5 Mins 0.0 0.5.0.5.0.5.0.5.0.5 Mins SIM : m/z=8 Peak Area : 00% SIM : m/z=8 Peak Area : 99% Column: ODP HP-B (50 x.0mm) Eluent:0mM CH COONH CH CN (70:0) Flow rate: 0.ml/min Column temperature: 0 C Detector: UV, 0nm and ESI-MS (-ve SIM) For free technical advice and support email technical@hichrom.co.uk 0.0 0.5.0.5.0.5.0.5.0.5 Mins Figure. LC-MS of barbital in BSA Ordering Information - ODP HP Product Particle Size (µm) Column Dimensions (mm) Catalogue No. Price ODP HP-B 5 50 x.6 F7600 06 ODP HP-D 5 50 x.6 F7600 0 ODP HP-E 5 50 x.6 F7600 5 ODP HPG-A 5 0 x.6 (guard column, /pk) F6700 ODP HP-GC 5 0 x.0 (guard cartridge, /pk) F670 87 ODP HP-B 5 50 x.0 F7600 06 ODP HP-D 5 50 x.0 F76005 0 ODP HPG-A 5 0 x.0 (guard column, /pk) F670 Use with guard cartridge holder HLD-GC (Cat. No. F870000, 0). 0.0 0.5.0.5.0.5.0.5.0.5 Mins Reversed-Phase (Silica-based) Phases Please enquire for details of reversed-phase silica-based columns, including Silica NPE (nitrophenylethyl) and Silica PYE (pyrenylethyl). 0

Please contact Hichrom for current prices. Tel: + (0)8 90 660 Email: sales@hichrom.co.uk Columns for Saccharide Analysis Ligand Exchange Phases ( SUGAR Series) SUGAR SH Styrene-divinylbenzene Sulpho (H + ) Separation of saccharides by ligand exchange, SUGAR SC Styrene-divinylbenzene Sulpho (Ca + ) size exclusion, ion exclusion and normal-phase. SUGAR SP Styrene-divinylbenzene Sulpho (Pb + ) Suitable for separation of sugars and sugar alcohols SUGAR SZ Styrene-divinylbenzene Sulpho (Zn + ) SUGAR KS Styrene-divinylbenzene Sulpho (Na + ) Separation of saccharides by size exclusion and ligand exchange USPpak MN Styrene-divinylbenzene Sulpho (Ca + ) Analysis of mannitol by USP method There are two series of columns for saccharide analysis the SUGAR series for operation in ligand exchange mode and Asahipak NHP-50 polymeric amino columns for normal-phase and HILIC modes. A. SUGAR Series The SUGAR series columns are based on polystyrene-divinylbenzene copolymer with strong cation-exchange resin incorporating functional sulpho groups coupled with metal counter ions (H +, Ca +, Pb +, Zn +, Na + ). B. Asahipak NHP Asahipak NHP is an amino bonded column in which polyamine is bonded to a hydrophilic polyvinyl alcohol polymer. The high separation power of conventional silica-based columns is maintained, whilst showing an increased usable ph range ( ) and increased robustness. Figure illustrates the reproducibility over time obtained from the polymeric Asahipak NHP column.. Fructose. Glucose. Sucrose. Maltose 80 h Column: Asahipak NHP-50 E (50 x.6mm) Eluent: CH CN H O (75:5) Flow rate:.0ml/min Column temperature: 0 C Detection: RI Figure. Reproducibility with Asahipak NHP-50 0 0 0 Mins 0 0 0 Mins Ordering Information - Columns for Saccharide Analysis Product Counter Separation Exclusion Limit Particle Size Column Dimensions Ion Mode (Da) (µm) (mm) Catalogue No. Price SUGAR SH0 H + SEC + IEX,000 6 00 x 8.0 F67800 859 SUGAR SH8 H + SEC + IEX 0,000 6 00 x 8.0 F6780 859 SUGAR SH-G H + - Guard column 0 50 x 6.0 F6700080 07 SUGAR SC0 Ca + SEC + LEX,000 6 00 x 8.0 F6780 859 SUGAR EP SC0-7F Ca + SEC + LEX,000 6 00 x 7.8 F67900 696 SUGAR SC8 Ca + SEC + LEX 0,000 6 00 x 8.0 F6780 859 SUGAR SC-LG Ca + - Guard column 0 50 x 6.0 F6700090 07 SUGAR SP080 Pb + SEC + LEX,000 7 00 x 8.0 F67805 859 SUGAR SP-G Pb + - Guard column 0 50 x 6.0 F670008 07 SUGAR SC Ca + NP + LEX - 6 50 x 6.0 F70000 859 SUGAR SC-G Ca + - Guard column 0 0 x.6 F67000 07 SUGAR SZ55 Zn + NP + LEX - 6 50 x 6.0 F70000 859 SUGAR SZ-G Zn + - Guard column 6 0 x.6 F67000 07 SUGAR KS-80 Na + SEC + LEX,000 6 00 x 8.0 F67800 90 SUGAR KS-80 Na + SEC + LEX 0,000 6 00 x 8.0 F67800 90 SUGAR KS-80 Na + SEC 50,000 6 00 x 8.0 F67805 90 SUGAR KS-80 Na + SEC 00,000 7 00 x 8.0 F67805 90 SUGAR KS-805 Na + SEC 5,000,000 7 00 x 8.0 F678050 90 SUGAR KS-806 Na + SEC 50,000,000 7 00 x 8.0 F678060 90 SUGAR KS-G Na + - Guard column 0 50 x 6.0 F670000 07 SUGAR KS-807 Na + SEC 00,000,000 0 00 x 8.0 F678070,6 SUGAR KS-807G Na + - Guard column 0 50 x 8.0 F67000 USPpak MN- Ca + LEX - 8 50 x.0 F6790 85 Asahipak Phases NHP-0 E - NP/HILIC - 50 x.0 F760007 7 NHP-50G A - - Guard column 5 0 x.0 F67000 8 NHP-50 D - NP/HILIC - 5 50 x.6 F76000 69 NHP-50 E - NP/HILIC - 5 50 x.6 F76000 70 NHP-50G A - - Guard column 5 0 x.6 F67006 76 NHP-50 D - NP/HILIC - 5 50 x.0 F760006 68 NHP-50G A - - Guard column 5 0 x.0 F67000 76 NHP-LF - - Line filter - 75 x 8.0 F67000 8 Preparative columns also available Hichrom Limited www.hichrom.co.uk sales@hichrom.co.uk Tel: 08 90 660

Please contact Hichrom for current prices. Tel: + (0)8 90 660 Email: sales@hichrom.co.uk Ion-Exchange Phases Ion-Exchange Phases IEC QA Polyhydroxymethacrylate Quaternary ammonium Strong anion-exchange IEC DEAE Polyhydroxymethacrylate Diethylaminoethyl Weak anion-exchange Suitable for analysis of proteins, peptides, DNA, IEC SP Polyhydroxymethacrylate Sulphopropyl Strong cation-exchange RNA, oligonucleotides IEC CM Polyhydroxymethacrylate Carboxymethyl Weak cation-exchange Asahipak ES-50N Polyvinyl alcohol Diethylaminoethyl Weak anion-exchange Asahipak ES-50C Polyvinyl alcohol Carboxymethyl Weak cation-exchange AXpak WA Polyhydroxymethacrylate Diethylaminoethyl Weak anion-exchange, suitable for analysis of nucleic acids CXpak P Styrene-divinylbenzene Sulpho (Na + ) Analysis of amino acids and amino sugars PIKESS DEAE Polyhydroxymethacrylate Diethylaminoethyl Weak anion-exchange UHPLC compatible PIKESS SP Polyhydroxymethacrylate Sulphopropyl Strong cation-exchange (.5µm material) The IEC series (QA and DEAE anion-exchange phases and SP and CM cation-exchange phases) is based on polyhydroxymethacrylate. Columns are suitable for the analysis of relatively high molecular weight compounds, such as proteins. The newer PIKESS materials are non-porous.5µm particles, suitable for rapid protein analyses using UHPLC systems (see Figure ).. Myoglobin. Trypsinogen. Ribonuclease A. a-chymotrypsinogen A 5. Cytochrome C 6. Lysozyme 5 6 Column: PIKESS SP-B (50 x.0mm) Eluent: A: 0mM Na phosphate, ph 7.0 B: A + 0.5M NaCl Linear gradient, 00% A to 50% A in.5min Flow rate:.ml/min Detection: UV, 80nm 5µ S/cm Column: IC YS-50 (5 x.6mm) Eluent: mm methanesulphonic acid Flow rate:.0ml/min Column temperature: 0 C Detector: Non-suppressed conductivity 5 6. Li + ppm. Na + 0ppm. NH + 0ppm. K + 0ppm 5. Mg + 0ppm 6. Ca + 0ppm 0.0.0.0.0.0 Mins Figure. Rapid analysis of proteins using UHPLC and ion-exchange 0.0 5 0 5 0 Mins Figure. Separation of cations using ion chromatography Please enquire for ordering details of ion-exchange columns. Ion Chromatography Phases Ion Chromatography Phases IC NI, I Polyhydroxymethacrylate Quaternary ammonium Anion analysis with non-suppressed detection IC SI Polyvinyl alcohol Quaternary ammonium Anion analysis with suppressed detection Cations by non-suppressed detection. Y for alkylamines. IC Y, T, R Styrene-divinylbenzene Sulpho T for transition metals and R for rare earth metals by post-column reaction IC YK Silica Carboxyl Monovalent and divalent cations by non-suppressed detection IC YS Silica Carboxyl Higher performance version of YK For free technical advice and support email technical@hichrom.co.uk A range of columns are available for both non-suppressed and suppressed ion chromatography. Columns are also supplied for analysing transition metal ions and rare earth metal ions using post-column reactions. Figure shows the separation of six common cations on a IC YS-50 column with non-suppressed conductivity detection. Ordering Information - Columns for Ion Chromatography Column Particle Size (µm) Column Dimensions (mm) Catalogue No. Price Guard Column Catalogue No. Price IC NI- 5 00 x.6 F6995 85 IC NI-G F670966 8 IC I-5A 00 x.6 F69950 767 IC IA-G F670000 8 IC Y-5 50 x.6 F69950 806 IC Y-G F67000 8 IC YK- 5 5 x.6 F700 708 IC YK-G F6709608 5 IC YS-50 5 5 x.6 F7000,06 IC YS-G F670050 76 IC SI-90 E 9 50 x.0 F6995 78 IC SI-90G F670960 8 IC SI-50 E 5 50 x.0 F69955 97 IC SI-50G F670965 8 IC T-5 50 x.6 F699550 979 IC T-G F6700 IC R-6 5 50 x 6.0 F6998000 806 IC R-G F6709090 8 PEEK hardware

Please contact Hichrom for current prices. Tel: + (0)8 90 660 Email: sales@hichrom.co.uk GFC Phases Aqueous SEC (GFC) Phases PROTEIN KW Silica Hydrophilic polymer OHpak SB Polyhydroxymethacrylate - Analysis of high MW biological fluids. Suitable for separating proteins of MW few thousand to few million Da Analysis of water-soluble samples and for molecular weight distribution PROTEIN KW-800 and KW00 are silica based series with high protein recovery rates. Whereas the KW-800 columns have the highest sample loading, the reduced particle size of KW00 columns enable - times greater sensitivity. Figure 5 illustrates the enhanced sensitivity achieved for a range of proteins with a KW00 column.. Blue dextran 000 0.mg/ml. g-globulin 0.8mg/ml. Ovalbumin 0.8mg/ml. Myoglobin 0.56mg/ml 5. Uridine 0.0mg/ml 5 Columns: (A) KW0.5-F (00 x.6mm) (B) PROTEIN KW-80.5 (00 x 8mm) Eluent: 50mM Na phosphate buffer, ph 7.0 + 0.M NaCl Flow rate: (A) 0.ml/min (B).0ml/min Detector: UV, 80nm Sample volume: 0µl (A) KW0.5-F (B) KW-80.5 5 0 5 0 Mins 5 0 5 0 Mins 5 Figure 5. Comparison between KW00 and KW-800 columns The OHpak SB-800 and SB00 series are based on polyhydroxymethacrylate and are used for the analysis of a wide range of molecular weight compounds. Ordering Information GFC Columns PROTEIN Particle Size Pore Size (Å) Exclusion Limit (Da) Column Dimensions Catalogue Product (µm) Average Maximum Pullulan Protein (mm) No. Price KW-80.5 5 50 00 60,000 50,000 00 x 8.0 F6989000 877 KW-80 5 00,000 70,000 700,000 00 x 8.0 F69890 877 KW-80 7 500,500 500,000,000,000 00 x 8.0 F69890 877 KW-G 7 Guard column 50 x 6.0 F6700 07 KW0.5-F 50 00 60,000 50,000 00 x.6 F69890 98 KW0-F 50 800 50,000 600,000 00 x.6 F69890 98 KW0-F 5 500,500 500,000,000,000 00 x.6 F69890 98 KW05-F 5 700,000,00,000 0,000,000 00 x.6 F69890 98 KW00G-A 5 Guard column 0 x.6 F6700 6 OHpak Product Particle Size (µm) Pore Size (Å) (Maximum) Exclusion Limit (Da) Pullulan Column Dimensions (mm) Catalogue No. SB-80 HQ 8 00,000 00 x 8.0 F6900 960 SB-80.5 HQ 6 00 0,000 00 x 8.0 F690 960 SB-80 HQ 6 800 00,000 00 x 8.0 F690 960 SB-80 HQ 0,000,000,000 00 x 8.0 F690 960 SB-805 HQ 7,000,000,000 00 x 8.0 F690 960 SB-806 HQ 5,000 0,000,000 00 x 8.0 F6905 960 SB-806M HQ 5,000 0,000,000 00 x 8.0 F6906 960 SB-G 0 Guard column 50 x 6.0 F67090 07 SB-807 HQ 5 0,000 500,000,000 00 x 8.0 F6908,79 SB-807G 5 Guard column 50 x 8.0 F6709 07 SB0-E 0 0,000 50 x.6 F69 960 SB0.5-E 6 00 0,000 50 x.6 F69 960 SB0-E 6 800 00,000 50 x.6 F69 960 SB0-E 7,000,000,000 50 x.6 F69 960 SB00G-A 7 Guard column 0 x.6 F6709 07 Price Hichrom Limited www.hichrom.co.uk sales@hichrom.co.uk Tel: 08 90 660

Other Phases Aqueous/Organic SEC Phases Asahipak GF Polyvinyl alcohol None Both water and organic solvents can be used. Suitable for samples with both hydrophilic and hydrophobic moieties. The Asahipak GF Series columns are based on polyvinyl alcohol and can be used with both aqueous and organic solvents. Please enquire for further details and ordering information for aqueous/organic SEC phases. Organic SEC (GPC) Phases GPC KF, K, KD, HFIP, LF, HT, UT, AT Styrene-divinylbenzene None KF-800 series shipped in THF. K-800 series shipped in CHCl. KD-800 series shipped in DMF suitable for assay of polar polymers. HFIP suitable for engineering plastics eg. PET, polyamides. HT, UT and AT series for higher temperature GPC. Please enquire for further details and ordering information for organic SEC (GPC) phases. Hydrophobic Interaction Phase HIC PH Polyhydroxymethacrylate Phenyl Analysis of proteins The HIC PH-8 column separates proteins without denaturation. It is applicable to samples after treatment of ammonium sulphate fractions. Organic Acids Phase RSpak KC Styrene-divinylbenzene Sulpho Analysis of organic acids by ion exclusion and reversed-phase modes RSpak KC columns are suitable for the analysis of organic acids by ion exclusion and reversed-phase modes. Affinity Phases AFpak 6 kinds Polyhydroxymethacrylate 6 kinds of ligand Purification of biological molecules depending on biochemical affinity The AFpak range of affinity columns consists of 6 different ligand materials, designed for specific applications. Chiral Separation Phases ORpak CDA, CDB, CDC Polyhydroxymethacrylate Cyclodextrin derivative Suitable for separation of optical isomers and structural isomers. CDA is a-cd, CDB is b-cd, CDC is g-cd ORpak CDBS Silica Cyclodextrin derivative CDBS is b-cd ORpak CRX Polyhydroxymethacrylate L-amino acid derivative Separation of amino acids by ligand exchange For free technical advice and support email technical@hichrom.co.uk ORpak CD phases consist of a-, b- or g-cyclodextrin bonded to polyhydroxymethacrylate base material or silica (a-cyclodextrin). ORpak CRX columns have an L-amino acid derivative bonded to polyhydroxymethacrylate and are suitable for the separation of optical isomers of amino acids and hydroxy acids. Please contact Hichrom for further information on any column and for ordering information for columns not listed.