Analytical HPLC Column Introduction

Transcription

1 hplc columns introduction Analytical HPLC Column Introduction Grace Key Column Families Single-Source Solution for Discovery to Recovery Applications HPLC is commonly used in a wide range of applications, including drug discovery and purification for the pharmaceutical and biotechnology industries, environmental analysis, forensics, petrochemical analysis, food, cosmetics, and vitamins. The combined premier product lines of Alltech, Davisil, Flexit, Grom, Jones Chromatography, Modcol, and Vydac create a single-source solution for HPLC columns and accessories from discovery to recovery. Our column families include reversed-phase, normal-phase, HILIC, ion-exchange, ion-exclusion, size-exclusion, and affinity stationary phases for small- and large-molecule separations, and our column formats maximize their performance. Our product range includes standard and custom columns for analytical separations, preparative phases for scale up, bulk packings that customers can pack in their own columns, and accessories to maintain separation ruggedness and quality. To help select the appropriate column for your application, we describe key column families and highlight unique phases within these families. Whether the most important factor is analysis speed, column bleed, ph stability, resolution, adjustable selectivity, or analyte molecular weight, Grace offers a column to suit your application. Column Families Suitable for Small Molecules (<000 Molecular Weight) VisionHT Ultra High-Pressure Columns,000psig pressure-rated columns, with sub µm media for high-efficiency, high-speed separations VisionHT columns are optimized for high throughput and ultra high-pressure LC applications. Mechanically stable.µm media and ultra-low volume hardware delivers new separation benefits with excellent stability. A variety of phases available. Differentiated Phases: C8, C0-B, C8-P, C8-HL, HILIC, Silica Specifications: Spherical silica, monomerically bonded, proprietary endcapping, 00, 0Å pore size Formats: Ultra High-Pressure Alltech Alltima HP Columns Premium quality, exceptionally stable columns, without phase bleed Alltech Alltima HP phases are made from high purity silica to eliminate peak tailing. Proprietary bonding eliminates the problem of column bleed for MS and ELSD detection. Full range of phases with ph stability from to 0. Differentiated Phases: C8 for classic reversed-phase separations, EPS for extended polar selectivity, C8 HiLoad for extra reversed-phase retention, C8 Amide with low bleed, and HILIC. Specifications: High purity spherical silica, monomerically bonded, endcapped, 00, 0, 90Å pore size Formats: Microbore, Expedite, Rocket, Solvent- Reducer, Analytical, Prep 70 Alltech Prevail Columns more info For information about Grace analytical hardware formats, see page. more info For information about prep columns, see pages 8 7. Reversed-phase columns for use with 00% organic to 00% aqueous mobile phases Prevail phases are designed for wettability with aqueous and organic mobile phases, and are especially useful for broad gradients. Retain highly polar analytes with aqueous mobile phases as well as hydrophobic analytes in organic mobile phases. Differentiated Phases: Carbohydrate ES, Organic Acid Specifications: Spherical silica, monomerically bonded, endcapped, 0Å pore size Formats: Microbore, Expedite, Rocket, Solvent- Reducer, Analytical, Prep

2 Analytical HPLC Column Introduction Column Families Suitable for Small Molecules (<000 Molecular Weight) (continued) Column Families Suitable for Large Molecules (>000 Molecular Weight) Alltech Platinum Columns Alternate retention and selectivity for polar and nonpolar analytes available in sub µm particle sizes Alltech Platinum phases have a controlled polar silica surface which provides enhanced polar selectivity (EPS) in addition to reversed-phase retention. C8 and EPS C8 columns are a complimentary duo to separate polar and nonpolar analytes. Differentiated Phases: C8, EPS C8 Specifications: Spherical silica, monomerically bonded, proprietary endcapping, 00Å pore size Formats: Microbore, Expedite, Rocket, Solvent- Reducer, Analytical, Prep Vydac MS Columns Next generation Vydac media from the leader in peptide and protein separations Vydac MS columns provide unique selectivity and exceptional protein recovery. They require less TFA in the mobile phase for good peak shape, which increases microbore sensitivity of peptides and proteins by reducing quenching. Vydac MS columns are also applicable to hydrophobic proteins and peptide mapping. Differentiated Phases: C8, C8, C Specifications: Spherical silica, polymerically and monomerically bonded, endcapped, 00Å pore size Formats: Capillary, Microbore, Expedite, Rocket, Solvent-Reducer, Analytical, Prep hplc columns introduction GraceSmart Columns Vydac ProZap Columns Classic reversed-phase and premium performance at an exceptional value GraceSmart HPLC phases use high purity silica. Which translates into symmetrical peaks for acids and bases, and predictable reversed-phase selectivity. Whether routine analysis or new method development, use individually tested GraceSmart columns to get premium performance at exceptional value. Differentiated Phases: C8 Specifications: Spherical silica, monomerically bonded, endcapped, 0Å pore size Format: Analytical High-speed, high-efficiency columns for fast protein analysis Short 0 and 0mm ProZap columns with.µm particles provide high-speed protein and life science separations. Sharp, efficient peaks maximize method sensitivity. Differentiated Phases: C8 Specifications: Sub µm spherical silica, proprietary bonding and endcapping, 00Å pore size Format: Expedite GraceAlpha Columns Vydac Everest Columns A New Silica Generation New silica from Grace with high porosity surface and dense core ideal for scale-up applications. While high porosity surface increases mass transfer results in increased column efficiencies and loading capacity, dense core and highly spherical shape yields mechanically robust particle. Differentiated Phases: C8, C8, and Silica Specifications: High purity spherical silica with high porosity surface and dense core Formats: Analytical, Prep For peptide separations, peptide mapping and proteomics applications High-capacity Everest C8 columns are recommended as the first choice in peptide separations. Everest offer high capacity to resolve complex samples such as protein digests. Differentiated Phases: C8 Specifications: Spherical silica, monomerically bonded, endcapped, 00Å pore size Formats: Capillary, Microbore, Expedite, Analytical

3 hplc columns introduction HPLC Column Selection A Comparison of Reversed-Phase Columns Based on the widely accepted work and data of Drs. Lloyd Snyder and John Dolan., Grace has developed this column selection tool for choosing reversed phase HPLC columns based on peak capacities and column selectivity of polar and nonpolar compounds. Typically, chromatographers choose HPLC columns by comparing physical characteristics, such as surface area and carbon load. Often, this does not provide enough information about selectivity or capacity for adequate column selection. This chart provides a reliable means of choosing HPLC columns based on acidic, basic, and hydrophobic character. The Snyder/Dolan column test procedure has been described in a series of publications. Based on retention data for a series of standard mixtures and the same separation conditions (0% acetonitrile/buffer; ph.8 and 7.0; C), reversed-phase columns are characterized by five column-selectivity parameters: hydrophobicity (H), steric interaction (S*), hydrogen-bond acidity A, basicity (B), and relative silanol ionization or cation-exchange capacity C. Here we have chosen to graphically highlight data for H, A, and C, with C results at ph 7.0. Hydrophobicity (H) is often the primary analyte interaction with reversed phase columns and indicates overall capacity. Secondary interactions are often polar interactions with basic analytes. The degree of unprotonated base interaction A and protonated base interaction C with the packing material is measured and represented here. Directions for Using the Column Chart The chart lists the columns in descending order of hydrophobic capacity (H). To find similar HPLC columns to test as back-up columns, follow these steps. ) Find the column you are currently using and note neighboring columns which have similar (H) capacity factors. ) Compare the values for interaction of polar compounds ( A and C ). If there is more than one choice for a back-up column, then compare your actual sample to the test probes. If your sample is nonpolar, then place more emphasis on hydrophobic values. If your sample is basic (polar), then pay special attention to A and C and determine if your sample will be protonated ( A) or unprotonated (C ) and place greater emphasis on one of these values. Key to Chart Hydrophobic Indicator A Hydrogen bonding Indicator ph.8 (Protonated under acidic conditions) C Cation Exchange Indicator ph 7.0 (Unprotonated under neutral conditions) Manufacturer Column Selectivity Parameters Manufacturer Column Selectivity Parameters ZirChrom ZirChrom -PBD C8 A C.8 Merck LiChrospher 00 RP-8 C A.006 YMC J Sphere H80 C8 C A. Bischoff ProntoSIL 0--C8 H C A.00 Restek Allure C8 C A.6 Shiseido CAPCELL C8 M G C A.00 Phenomenex Ultracarb ODS (0) C A. Grace (Jones) Genesis 0 C8 C A.00 YMC YMC Pack Pro C8 RS C A. Bischoff EU Reference Column C8 C A.00 Grace (Alltech) Adsorbosphere UHS C8 C A.0 Grace (Alltech) Allsphere ODS CA.00 Thermo/Hypersil Hypersil BetamaxNeutral C8 C A.098 Merck Purospher STAR RP8e C A.00 Agilent Zorbax Extend C8 C A.098 Merck Chromolith RP8e C A.00 Agilent Zorbax C8 C A.089 Phenomenex Luna C8() C A.00 Beckman Ultrasphere ODS C A.08 Varian Pursuit C8 C A.00 Grace (Alltech) Alltima HP C8 High Load C A.080 MacMod/ACT ACE C8 C A.000 Agilent Zorbax Rx-8 C A.077 Tosoh TSKgel Super-ODS C A Agilent Zorbax Eclipse XDB-C8 C A.077 Agilent Zorbax StableBond 80A C8 C A Supelco Ascentis C8 C A.077 Phenomenex Prodigy ODS() C A 0.99 Macherey Nagel Nucleodur C8 Gravity C A.06 Thermo/Hypersil Hypersil BDS C8 C A 0.99 Grace (Grom) Grom Sapphire 0 C8 C A.0 Grace (Alltech) Alltima C8 C A 0.99 Restek Restek Ultra C8 C A.0 Grace (Vydac) Vydac Everest C8 C A 0.99 Varian OmniSpher C8 C A.0 Thermo/Hypersil Hypersil Beta Basic-8 C A 0.99 Grace (Vydac) Denali 0 C8 C A.0 GL Sciences Inertsil ODS- C A Waters Symmetry C8 C A.0 Grace (Alltech) Adsorbosphere C8 A C Akzo Nobel Kromasil 00-C8 C A.0 Phenomenex Synergi Max-RP C8 C A Waters Nova-Pak C8 C A.09 Shiseido CAPCELL C8 SG0 C A Thermo/Hypersil Hypersil 00 C8 C A.08 Grace (Jones) Apex I C8 A C 0.98 MacMod/ACT ACE C8-HL C A.0 Thermo/Hypersil Hypersil ODS- C A 0.98 ZirChrom ZirChrom -EZ C8 A C.00 Grace (Alltech) Alltima HP C8 C A 0.98 Grace (Grom) Grom Sil 0 ODS- ST C A.0 Waters Xterra MS C8 C A 0.98 Dionex Acclaim 0 C8 C A.0 Waters Symmetry 00 C8 C A 0.98 Waters Sunfire C8 C A.0 Supelco Discovery C8 C A 0.98 Agilent Zorbax Eclipse Plus C8 C A.00 Supelco Supelcosil LC-8-DB C A Merck Superspher 00 RP-8e C A.00 Waters Spherisorb S ODSB A C 0.97 Shiseido CAPCELL C8 AG0 C A.00 Thermo/Hypersil Hypersil Bio Basic-8 C A 0.97 Grace (Grom) Grom Sil 0 ODS- CP C A.09 Thermo/Hypersil Hypersil ODS A C 0.97 Waters Delta-Pak C8 00A C A.08 Bischoff ProntoSIL 0--C8-AQ C A 0.97 Macherey Nagel Nucleodur Isis C A.0 Grace (Jones) Genesis 00 C8 C8 C A 0.97 Phenomenex Prodigy ODS () C A.0 Phenomenex Synergi Hydro-RP C8 C A.0 Phenomenex Luna C8 C A.08 Supelco Supelcosil LC-8 A C.08 YMC YMC Pro C8 C A.0 Phenomenex Onyx Monolithic C8 C A.0 Bischoff ProntoSIL SpheriBOND 80--ODS C A.00 Grace (Jones) Apex II C8 A C.008 Shiseido CAPCELL C8 UG0 C A.007 GL Sciences Inertsil ODS- C A.007 Bischoff Prontosil 00--C8 AQ C A 0.97 Agilent Zorbax C8 C A 0.97 Tosoh TSK gel ODS-80Ts A C 0.97 Waters Resolve C8 A C Phenomenex Gemini C8 0A C A Grace (Alltech) Econosil C8 C A Phenomenex Aqua C8 C A YMC YMC ODS-AQ C8 C A 0.96 Waters Spherisorb ODS- AC 0.96 Macherey Nagel Nucleosil 00--C8 HD C8 C A

4 Grace (Jones) Genesis 0 AQ C8 C A Macherey Nagel Nucleodur Pyramid C A 0.98 Thermo/Hypersil Hypersil Elite C8 C A 0.98 Dionex Acclaim 00 C8 C A 0.97 Bischoff ProntoSIL 00--C8 H C A 0.96 Waters Delta-Pak C8 00A C A 0.9 Bischoff ProntoSIL HyperSORB 0 ODS A C 0.9 Thermo/Hypersil Hypersil PAH C8 A C 0.99 Bischoff ProntoSIL 0--C8 Aqplus C A 0.97 Phenomenex Jupiter 00 C8 C A 0.9 Varian Polaris C8-Ether 0.9 CA Waters Atlantis T C8 AC 0.9 Tosoh TSKgel 80Ts QA C A 0.90 Grace (Alltech) Alltima C8-WP C A 0.98 Waters YMC Hydrosphere C8 C A 0.97 Grace (Alltech) Brava BDS C8 C A 0.98 Bischoff ProntoSIL 60- C8 SH 0.99 C A Varian Polaris C8-A C A 0.98 YMC J Sphere M80 C8 C A 0.96 Agilent Zorbax Eclipse XDB-C8 C A 0.99 Waters Atlantis dc8 b C A 0.97 Supelco Ascentis Express C8 C A 0.9 Thermo Hypersil GOLD aq 0.9 C A Phenomenex Selectosil C8 C A 0.9 Merck LiChrosorb RP-8 C A Grace (Vydac) Vydac 8TP C8 C A Waters Acquity UPLC BEH Shield RP8 EP C A Macherey Nagel Nucleosil C C A Agilent Zorbax StableBond 00A C8 C A 0.90 Grace (Alltech) Prosphere C8 00 AC 0.90 Grace (Vydac) Vydac 0TP C8 C A 0.90 Supelco Ascentis C-8 C A Waters Nova-Pak C CA Waters Symmetry C8 C A 0.89 YMC YMC Pro C8 C A Agilent Zorbax Eclipse Plus C8 C A Phenomenex Luna C8() C A Grace (Alltech) Prevail C8 C A Grace (Alltech) Prosphere 00 C8 AC 0.88 Grace (Alltech) Alltima AQ EP C A 0.88 Thermo/Hypersil Hypersil GOLD C8 C A 0.88 Phenomenex Synergi Fusion-RP EP C A Phenomenex Luna C8 C A 0.87 Grace (Grom) Grom Sil 0 Octyl-6 MB C8 C A 0.87 Grace (Jones) Apex I C Shiseido CAPCELL C8 A Q C A Macherey Nagel Nucelosil 00--C8 HD C A 0.86 Akzo Nobel Kromasil 00-C8 C A 0.86 Grace (Jones) Genesis 0 EC C8 C A 0.86 Grace (Alltech) Prevail Amide EP C A 0.86 Macherey Nagel Nucleosil ODS AC Restek Ultra AQ C8 C A 0.87 Waters Sunfire C8 C A 0.86 Waters Acquity UPLC BEH C8 C A 0.8 Shiseido CAPCELL PAK C8 UG0 C A 0.8 Waters Symmetry Shield C8 AC 0.80 Grace (Alltech) Alphabond C8 A C 0.8 Supelco Ascentis RP-Amide AC 0.8 Merck Purospher RP-8 C A 0.8 Supelco Discovery BIO Wide pore C8 C A 0.89 Grace (Grom) Grom Sapphire 0 C8 C A 0.8 Thermo/Hypersil Hypersil Beta Basic-8 C A 0.8 Grace (Alltech) Alltima HP C8 C A 0.8 Thermo/Hypersil Hypurity C8 C A 0.8 Supelco Discovery C8 C A 0.8 GL Sciences Inertsil C8- C8 C A 0.80 MacMod/ACT ACE C8 C A 0.80 Grace (Jones) Genesis 0 C8 C A 0.89 Thermo Hypersil GOLD C8 C A 0.8 Phenomenex Onyx Monolithic C8 C A 0.8 Tosoh TSKgel Super-Octyl C A 0.8 Grace (Alltech) Prevail Select C8 AC 0.8 Thermo/Hypersil Hypersil Bio Basic-8 C 0.8 YMC YMC Basic C8 C A 0.8 Grace (Alltech) Econosphere C8 A C 0.88 Tosoh TSKgel Octyl-80Ts AC 0.8 Whatman Partisil ODS() AC 0.80 A AC Macherey Nagel Nucleodur Sphinx RP AC 0.80 Thermo/Hypersil Aquasil C8 C A 0.80 MacMod/ACT ACE AQ EP C A 0.80 Waters Xterra MS C8 C A 0.80 Phenomenex Luna C C A Waters MicroBondapak C8 AC Agilent Zorbax StableBond 80A C C A Agilent Zorbax Rx-C C A Grace (Alltech) Platinum C8 AC Grace VisionHT C8 AC Phenomenex Luna Phenyl-Hexyl AC 0.78 Grace (Alltech) Alltima C8-LL C A Bischoff ProntoSIL 0--C8 ace-eps A C 0.77 Grace (Vydac) Vydac 8MS C8 AC Waters Acquity UPLC BEH phenyl 0.76 C A Waters Spherisorb C8 AC 0.76 YMC J Sphere L80 C C A Bischoff ProntoSIL 00--C8 ace-eps A C 0.76 Dionex Acclaim Organic Acid C8 AC 0.76 Waters Xterra C8 RP 0.77 C A Grace (Alltech) Alltima C C A Whatman Partisil C C A Merck LiChrospher 60 RP-Select B C8 AC 0.77 Bischoff ProntoSIL 0- C8 SH C A 0.79 Grace (Alltech) Allsphere ODS C A 0.7 Waters Symmetry Shield C8 A C 0.70 Thermo/Hypersil Hypurity C 0.7 CA MacMod/ACT ACE C-00 C A 0.70 Varian Polaris C8-Ether AC 0.70 Bischoff ProntoSIL 60--Phenyl A C 0.70 Waters Nova-Pak Phenyl A C 0.70 Macherey Nagel Nucleosil 00--C8 Nautilus A C 0.70 Agilent Zorbax StableBond 00A C CA Bischoff ProntoSIL SpheriBOND 80--ODS A C Thermo/Hypersil Fluophase RP F AC Phenomenex Jupiter 00 C AC Grace (Alltech) Prosphere 00 C CA Bischoff Prontosil 60--C AC Waters Xterra Phenyl 0.68 C A Waters Spherisorb ODS- C A 0.68 Thermo/Hypersil Hypersil Prism C8 RPN C A Thermo/Hypersil Fluophase PFP F A C 0.67 Agilent Zorbax XDB-Phenyl AC 0.66 Waters Symmetry 00 C AC 0.69 Waters Xterra C8 RP AC 0.67 Grace (Alltech) Alltima HP C8 EPS C A 0.6 Supelco Discovery BIO Wide pore C 0.6 CA Agilent Zorbax Bonus RP EP A C 0.6 Phenomenex Synergi Polar-RP C8 AC 0.6 MacMod/ACT ACE Phenyl AC 0.67 Grace (Jones) Genesis 0 C EC AC 0.66 Thermo/Hypersil Hypersil Prism C8 RP C A 0.6 Thermo/Hypersil BetaMax Acid EP 0.6 C A Supelco Discovery HS F F A C 0.6 Agilent Zorbax SB-Phenyl AC 0.6 Grace (Alltech) Platinum EPS C8 AC 0.69 Grace VisionHT C8-P AC 0.69 Grace (Alltech) Prevail C8 AC 0.67 Grace (Jones) Genesis 00 C C AC 0.6 Grace (Jones) Genesis Phenyl A C Agilent Zorbax StableBond 80A C AC 0.60 MacMod/ACT ACE Phenyl-00 AC 0.99 Agilent Zorbax SB-AQ EP AC 0.9 ZirChrom ZirChrom -PS EP A C 0.89 Waters MicroBondapak Phenyl A C 0.8 Grace (Alltech) Platinum C8 0.8 CA Grace (Alltech) Platinum EPS C8 00 A C 0.8 Thermo/Hypersil BetaBasic Phenyl A C 0.8 Macherey Nagel Nucleosil C8 AC 0.7 Macherey Nagel EC Nucleosil 00- Protect EP C A 0. Bischoff Prontosil 0--C8 ace-eps A C 0. Phenomenex Prodigy Phenyl- A C 0.9 Agilent Zorbax StableBond 00A C 0.6 C A Grace (Alltech) Alltima HP C8 Amide C A 0.97 Thermo/Hypersil BetaMax Base EP A C 0.70 Grace (Alltech) Platinum EPS C8 AC 0.0 Thermo/Hypersil Hypurity Advance AC 0. References:. The Hydrophobic-subtraction Model of Reversed-phase Column Selectivity, L.R. Snyder, J.W. Dolan and P.W. Carr, J. Chromatogr. A, 060 (00) A New Look at the Selectivity of Reversed-phase HPLC Columns, L.R. Snyder, J.W. Dolan and P.W. Carr, Anal. Chem., 79 (007) -6. hplc columns introduction 7

5 hplc columns introduction 8 Grace HPLC Packing Material Specifications Columns for Small Molecules Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Endcapped? USP L-code Brand Phase Phase Type Adsorbosil C8 Silica Irregular, 0µm 60Å 0m /g % Polymeric Yes L C8 Silica Irregular, 0µm 60Å 0m /g 0% Polymeric Yes L7 C Silica Irregular, 0µm 60Å 0m /g No Polymeric No L6 CN Silica Irregular, 0µm 60Å 0m /g Polymeric Yes L0 NH Silica Irregular, 0µm 60Å 0m /g Polymeric No L8 Silica Silica Irregular, 0µm 60Å 0m /g Polymeric No L Adsorbosphere C8 Silica Spherical,, 0µm 80Å 00m /g % Monomeric Yes L C8 HS Silica Spherical, µm 60Å 0m /g 0% Monomeric Yes L C8 UHS Silica Spherical, 0µm 60Å 00m /g 0% Monomeric Yes L C8 Silica Spherical,, 0µm 80Å 00m /g 8% Monomeric Yes L7 Phenyl Silica Spherical µm 80Å 00m /g % Monomeric Yes L Cyano Silica Spherical µm 80Å 00m /g Monomeric Yes L0 Cyano-AQ Silica Spherical µm 0Å 70m /g Polymeric No L0 Amino (NH) Silica Spherical, µm 80Å 00m /g Polymeric No L8 Silica Silica Spherical µm 80Å 00m /g No L SAX Silica Spherical µm 80Å 00m /g Monomeric No SCX Silica Spherical µm 80Å 00m /g Monomeric Yes Adsorbosphere XL C8 Silica Spherical, µm 90Å 00m /g % Monomeric Yes L C8-B Silica Spherical µm 90Å 00m /g % Monomeric Yes L C8 Silica Spherical, µm 90Å 00m /g 6% Monomeric Yes L7 C (TMS) Silica Spherical µm 90Å 00m /g Monomeric Yes L Silica Silica Spherical µm 90Å 00m /g No L SAX Silica Spherical, 0µm 90Å 00m /g Monomeric Yes SCX Silica Spherical, 0µm 90Å 00m /g Monomeric Yes Allsphere ODS- Silica Spherical µm 80Å 0m /g 7% Monomeric Partial L ODS- Silica Spherical, µm 80Å 0m /g % Monomeric Yes L C8 Silica Spherical, µm 80Å 0m /g 6% Monomeric Yes L7 C6 Silica Spherical µm 80Å 0m /g % Monomeric Yes L C (TMS) Silica Spherical µm 80Å 0m /g % Monomeric No L Phenyl Silica Spherical µm 80Å 0m /g % Monomeric Yes L Cyano Silica Spherical µm 80Å 0m /g.% Monomeric No L0 Amino (NH) Silica Spherical µm 80Å 0m /g % Monomeric No L8 Silica Silica Spherical, µm 80Å 0m /g No L SAX Silica Spherical µm 00Å 0m /g % Monomeric No SCX Silica Spherical µm 00Å 0m /g % Monomeric No Alltima HP C8 Silica Spherical, µm 90Å 00m /g % Monomeric Yes L C8 EPS Silica Spherical, µm 90Å 00m /g % Monomeric No L C8 HiLoad Silica Spherical, µm 00Å 0m /g % Monomeric Yes L C8 AQ Silica Spherical, µm 00Å 0m /g 0% Monomeric Yes L C8 Amide Silica Spherical, µm 90Å 00m /g % Monomeric Yes L C8 Silica Spherical, µm 90Å 00m /g 8% Monomeric Yes L7 Cyano Silica Spherical, µm 90Å 00m /g % Monomeric Yes L0 Silica Silica Spherical, µm 00Å 0m /g No L HILIC Silica Spherical.,, µm 0Å 0m /g No L Alltima C8 Silica Spherical,, 0µm 00Å 0m /g 6% Polymeric Yes L C8 LL Silica Spherical µm 00Å 0m /g 9% Polymeric Yes L C8 Silica Spherical,, 0µm 00Å 0m /g 9% Polymeric Yes L7 Phenyl Silica Spherical, µm 00Å 0m /g 7.% Polymeric Yes L Cyano Silica Spherical, µm 00Å 0m /g Polymeric Yes L0 Amino (NH) Silica Spherical, µm 00Å 0m /g Polymeric No L8 Silica Silica Spherical,, 0µm 00Å 0m /g No L AlphaBond C8 Silica Irregular, 0µm Å 00m /g 0% Monomeric Yes L C8 Silica Irregular 0µm Å 00m /g Monomeric Yes L7 Phenyl Silica Irregular 0µm Å 00m /g Monomeric Yes L Cyano Silica Irregular 0µm Å 00m /g Monomeric Yes L0 Amino (NH) Silica Irregular 0µm Å 00m /g Polymeric No L8 Silica Silica Irregular 0µm Å 00m /g No L Apex I ODS Silica Spherical,, 0µm 00Å 70m /g 0% Polymeric Yes L C8 Silica Spherical, µm 00Å 70m /g 7% Monomeric No L7 C8(EC) Silica Spherical, µm 00Å 70m /g 7% Monomeric Yes L7 C Silica Spherical, µm 00Å 70m /g.% Monomeric Yes L Phe Silica Spherical, µm 00Å 70m /g % Monomeric No L Basic ODS Silica Spherical µm 00Å 00m /g % Monomeric Yes L PAH Silica Spherical µm 00Å 70m /g Monomeric Yes CN Silica Spherical, µm 00Å 70m /g % Monomeric No L0 Amino (NH) Silica Spherical, µm 00Å 70m /g % Monomeric No L8 Carbohydrate Silica Spherical µm 00Å 70m /g Monomeric Silica Silica Spherical,, 0µm 00Å 70m /g No L Apex II ODS Silica Spherical, µm 00Å 70m /g 0.% Monomeric Yes L Diol Silica Spherical, µm 00Å 70m /g.% Monomeric No Amino (NH) Silica Spherical, µm 00Å 70m /g % Monomeric No L8 Others as Apex I Silica Spherical µm 00Å 70m /g No Apex Prepsil ODS Silica Spherical 8, µm 0Å 70m /g 0% Polymeric Yes L C8 Silica Spherical 8, µm 0Å 70m /g 7% Monomeric No L7 C8(EC) Silica Spherical 8, µm 0Å 70m /g 7% Monomeric Yes L7 C Silica Spherical 8µm 0Å 70m /g.8% Monomeric No L0 CN Silica Spherical 8µm 0Å 70m /g % Monomeric Yes L0 Amino (NH) Silica Spherical 8µm 0Å 70m /g % Monomeric Yes L8 Silica Silica Spherical 8, µm 0Å 70m /g No L Diol Silica Spherical 8µm 0Å 70m /g.% Monomeric No L0 Proprietary.

6 Columns for Small Molecules (continued) Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Endcapped? USP L-code Brand Phase Phase Type Apollo C8 Silica Spherical µm 00Å 0m /g % Monomeric Yes L C8 Silica Spherical µm 00Å 0m /g 9% Monomeric Yes L7 Phenyl Silica Spherical µm 00Å 0m /g 8% Monomeric Yes L Silica Silica Spherical µm 00Å 0m /g No L Brava C8 BDS Silica Spherical, µm Å 8m /g 8.% Monomeric Yes L C8 ODS Silica Spherical, µm 0Å 9m /g 8.% Monomeric Yes L C8 Silica Spherical, µm 0Å 9m /g 6% Monomeric Yes L7 C8 BDS Silica Spherical, µm Å 8m /g.% Monomeric Yes L7 Phenyl Silica Spherical µm 0Å 9m /g Monomeric No L Cyano Silica Spherical µm 0Å 9m /g Monomeric No L0 Cyano BDS Silica Spherical µm Å 8m /g Monomeric No L0 Amino (NH) Silica Spherical µm 0Å 9m /g Monomeric No L8 Silica Silica Spherical µm 0Å 9m /g No L Carbohydrate Amino Silica Irregular 0µm 80Å 0m /g Polymeric No Cation Polymer Spherical 0µm No Denali 8DE C8 Silica Spherical,, 0,, 0µm 0Å 80-0m /g 0% Monomeric Yes L Econosil * C8 Silica Irregular,, 0µm 80Å 00m /g 0% Monomeric Yes L C8 Silica Irregular,, 0µm 80Å 00m /g % Monomeric Yes L7 CN Silica Irregular, 0µm 80Å 00m /g Monomeric Yes L0 NH Silica Irregular, 0µm 80Å 00m /g Polymeric No L8 Silica Silica Irregular,, 0µm 80Å 00m /g No L Econosphere * C8 Silica Spherical,, 0µm 80Å 00m /g 0% Monomeric Yes L C8 Silica Spherical,, 0µm 80Å 00m /g % Monomeric Yes L7 Cyano Silica Spherical µm 80Å 00m /g Monomeric Yes L0 Amino (NH) Silica Spherical µm 80Å 00m /g Polymeric No L8 Silica Silica Spherical,, 0µm 80Å 00m /g No L Genesis 0 C8 Silica Spherical,, 7, µm 0Å 00m /g 8% Monomeric Yes L C8 AQ Silica Spherical, 7µm 0Å 00m /g % Monomeric Yes L C8 Silica Spherical,, 7, µm 0Å 00m /g % Monomeric No L7 C8(EC) Silica Spherical,, 7, µm 0Å 00m /g % Monomeric Yes L7 C Silica Spherical µm 0Å 00m /g 6% Monomeric Yes L6 Phenyl Silica Spherical µm 0Å 00m /g 9% Monomeric Yes L CN Silica Spherical, µm 0Å 00m /g 7% Monomeric Yes L0 Amino (NH) Silica Spherical, µm 0Å 00m /g.% Polymeric No L8 Carbohydrate Silica Spherical µm 0Å 00m /g Monomeric No CN-TCA Silica Spherical µm 0Å 00m /g 7% Monomeric Yes Petro-XP Silica Spherical µm 0Å 00m /g Monomeric No Silica Silica Spherical,, 7, µm 0Å 00m /g No L Grace Alpha C8 Silica Spherical, 0,, 0µm 0Å m /g % Monomeric Yes L C8 Silica Spherical, 0,, 0µm 0Å m /g 0% Monomeric No L7 Silica Silica Spherical, 0,, 0µm 0Å m /g No L GraceSmart C8 Silica Spherical, µm 0Å 0m /g 0% Monomeric Yes L Grom Sil ODS-0 AB (acid/base deactivated) Silica Spherical.,,, 0µm 00Å 00m /g % Monomeric Yes L ODS- FE (fully endcapped) Silica Spherical.,,, 0µm 80, 00, 00Å 0, 00, 00m /g,, 6% Monomeric Yes L ODS- CP (encapsulated) Silica Spherical,, 7, 0µm 0, 00Å 0, 70m /g, 6% Polymeric No L ODS- HE (hydrophilic endcapping) Silica Spherical,,, 7, 0µm 0, 00Å 00, 00m /g 6, % Monomeric Yes L ODS- ST (standard) Silica Spherical,,, 7, 0µm 60, 0, 00, 00Å 80, 00, 00, 0m /g, 7,, 7% Monomeric Yes L ODS-6 NE (non endcapped) Silica Irregular, µm 0Å 00m /g 7% Monomeric No L ODS-7 ph (ph-stable) Silica Irregular µm 80Å 0m /g % Polymeric No L Octyl- B (base deactivated) Silica Spherical, µm 00Å 00m /g 6.% Monomeric Yes L7 Octyl- AB (acid/base deactivated) Silica Spherical, µm 00Å 00m /g % Monomeric Yes L7 Octyl- BA (for bases) Silica Spherical, µm 0Å 00m /g 9% Monomeric Yes L7 Octyl- FE (fully endcapped) Silica Spherical,, 0µm 80, 00, 00Å 0, 00, 00m /g 6.6, 6, % Monomeric Yes L7 Octyl- CP (encapsulated) Silica Spherical,, 7, 0µm 0, 00Å 0, 70m /g 0,.% Polymeric No L7 Octyl-6 MB (monomer binding) Silica Spherical,, 0µm 0, 00, 00Å 00, 00, 0m /g 0, 7, % Monomeric Yes L7 Hexyl- MB (monomeric bonding) Silica Spherical µm 80, 00Å 0, 00m /g, % Monomeric Yes Phenyl- FE (fully endcapped) Silica Spherical,, 0µm 0, 00Å 00, 0m /g 9, % Monomeric Yes L Phenyl- CP (encapsulated) Silica Irregular µm 0, 00Å 0, 70m /g 7, % Polymeric No L Phenyl- PE (partially endcapped) Silica Spherical,, 0µm 80, 00Å 0, 00m /g 6.6, 6% Monomeric Yes L Butyl- ST (standard) Silica Spherical, µm 0, 00Å 00, 0m /g 7,.% Monomeric No L6 Butyl- FE (fully endcapped) Silica Spherical, µm 00Å 00m /g, % Monomeric No L6 TMS- ST (standard) Silica Spherical, µm 0, 00Å 00, 0m /g % Monomeric Yes L TMS- CP (encapsulated) Silica Spherical, µm 0, 00Å 0, 70m /g % Polymeric No L Cyan- ST (standard) Silica Spherical, µm 0, 00Å 00, 0m /g.8% Monomeric Yes Cyan- PR (cyanopropyl) Silica Spherical, µm 80, 00Å 0, 00m /g.% Monomeric Yes Cyan- CP (encapsulated) Silica Spherical µm 0Å 0m /g % Polymeric No Amino- PR (NH-propyl) Silica Spherical,, 0µm 80, 00Å 0, 00m /g % Monomeric Yes L8 Amino- PA (cross linked Poly-NH) Silica Spherical µm 0Å 00m /g Polymeric No L8 Amino- CP (encapsulated NH-residues) Silica Irregular µm 80Å 0m /g Monomeric Yes L8 Amino- PR (propylamine bonded to silica) Silica Irregular, 7µm 00Å 00m /g Monomeric No L8 Diol Silica Spherical, 0µm 60, 0, 00, 00Å 80, 00, 00, 0m /g Monomeric No L0 Normal Phase- ST (standard silica) Silica Spherical,, 0µm 80, 00, 000Å 0, 00m /g No L Normal Phase- SP (spherical silica) Silica Spherical,, 0µm 60, 0, 00, 80, 00, 00m /g No L 000Å Normal Phase- PV (polyvinylalcohol) Silica Spherical µm 0Å 00m /g Polymeric No L SEC (size exclusion chromatography) Silica Spherical, 0µm 60, 0, 00, 00Å 80, 00, 00, 0m /g No Strong Anion- Silica Spherical, 0µm 80, 00Å 0, 00m /g No Weak Anion- (ion exchange) Silica Spherical 7µm 00Å 00m /g No Strong Cation- (ion exchange) Silica Spherical, 0µm 80, 00Å 0, 00m /g No *Available only online. hplc columns introduction 9

7 hplc columns introduction Columns for Small Molecules (continued) Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Endcapped? USP L-code Brand Phase Phase Type Grom Sil (cont.) Weak Cation- (ion exchange) Silica Spherical 7µm 00Å 00m /g No HIC (hydrophobic interaction chrom.) Silica Spherical 7µm 00Å 00m /g No Grom Sapphire C8 Silica Spherical,, 0µm 6, 0Å 00, 70m /g, 6% Monomeric Yes L C8 Silica Spherical,, 0µm 6, 0Å 00, 70m /g, 0% Monomeric Yes L7 C Silica Spherical,, 0µm 6, 0Å 00, 70m /g 0., 7% Monomeric Yes L6 Silica Silica Spherical,, 0µm 6, 0Å 00, 70m /g No L Mixed Mode C8/Cation Silica Spherical, 7µm 00Å 0m /g Polymeric No C8/Anion Silica Spherical 7µm 00Å 0m /g Polymeric No C8/Cation Silica Spherical µm 00Å 0m /g Polymeric No Platinum C8 Silica Spherical.,, µm 00Å 00m /g 6% Monomeric Yes L C8 EPS Silica Spherical.,, µm 00Å 00m /g % Monomeric No L C8 Silica Spherical.,, µm 00Å 00m /g % Monomeric Yes L7 C8 EPS Silica Spherical, µm 00Å 00m /g.% Monomeric No L7 Phenyl Silica Spherical, µm 00Å 00m /g Monomeric Yes L Cyano Silica Spherical, µm 00Å 00m /g Monomeric No L0 Amino (NH) Silica Spherical, µm 00Å 00m /g Monomeric No L8 Silica Silica Spherical, µm 00Å 00m /g No L SAX Silica Spherical, µm 00Å 00m /g Monomeric No Prevail C8 Select Silica Spherical, µm 0Å 0m /g 7% Monomeric Yes L C8 Silica Spherical, µm 0Å 0m /g % Monomeric Yes L C8 Silica Spherical, µm 0Å 0m /g 8% Monomeric Yes L7 Phenyl Silica Spherical, µm 0Å 0m /g 7% Monomeric Yes L Cyano Silica Spherical, µm 0Å 0m /g Monomeric Yes L0 Amino (NH) Silica Spherical, µm 0Å 0m /g Monomeric No L8 Silica Silica Spherical, µm 0Å 0m /g No L Organic Acid Silica Spherical, µm 0Å 0m /g Monomeric Yes Carbohydrate ES Polymer Spherical µm No Vydac SP 0SP Sil Silica Spheroidal, 0µm 00Å 0 0m /g unbonded No L 0SP C8 Silica Spheroidal,, 0, µm 90Å 0 0m /g % Monomeric Yes L 08SP C8 Silica Spheroidal, 0, µm 90Å 0 0m /g 9% Monomeric Yes L7 VisionHT C8 Silica Spherical.µm 00Å 00m /g 6% Monomeric Yes L C-8-B Silica Spherical.µm 0Å 0m /g.% Monomeric L C8-P Silica Spherical.µm 00Å 00m /g % Monomeric No L C8-HL Silica Spherical.µm 0Å 0m /g % Polymeric L HILIC, Silica Silica Spherical.µm 0Å 0m /g No L Columns for Large Molecules Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Endcapped? USP L-code Brand Phase Phase Type Genesis 00 C8 Silica Spherical, 7µm 00Å 0m /g 0% Monomeric Yes L C Silica Spherical, 7µm 00Å 0m /g % Monomeric Yes L6 CN Silica Spherical µm 00Å 0m /g.% Monomeric Yes L Macrosphere 00 C8 Silica Spherical, 7µm 00Å 00m /g 0% Monomeric Yes L C8 Silica Spherical, 7µm 00Å 00m /g.% Monomeric Yes L7 C Silica Spherical, 7µm 00Å 00m /g Monomeric Yes L6 SAX Silica Spherical 7µm 00Å 00m /g Monomeric No WAX Silica Spherical 7µm 00Å 00m /g Monomeric No SCX Silica Spherical 7µm 00Å 00m /g Polymeric No WCX Silica Spherical 7µm 00Å 00m /g Polymeric No Macrosphere GPC GPC 60 Silica Spherical 7µm 60Å 0m /g Polymeric No L GPC 00 Silica Spherical 7µm 00Å 0m /g Polymeric No GPC 0 Silica Spherical 7µm 0Å 00m /g Polymeric No GPC 00 Silica Spherical 7µm 00Å 00m /g Polymeric No ProSphere C8 Silica Spherical,, 0µm 00Å 0m /g 9% Monomeric Yes L C8-AQ Silica Spherical, µm 00Å 0m /g 8% Monomeric Yes L C Silica Spherical,, 0µm 00Å 0m /g % Monomeric Yes L6 Size-Exclusion Silica Spherical, µm Å No Size-Exclusion 0 Silica Spherical, µm 0Å No Size-Exclusion 0 Silica Spherical 8µm 0Å No P-HR (reversed phase) Polymer Spherical µm 0Å No ProZap C8 ProZap Silica Spherical.µm 00Å 9m /g % Monomeric Yes L Vydac ATP ATP C Silica Spheroidal, 0 µm 00Å 70 0m /g % Polymeric Yes L6 Vydac TP 0TP Sil Silica Spheroidal, 0, 0, 0µm 00Å 70 0m /g unbonded No L 0TP C8 Silica Spheroidal, 7, 0, 0, 00Å 70 90m /g 8% Polymeric No L 0µm 0TP C8 Silica Spheroidal,, 0µm 00Å 60 90m /g 9% Polymeric No L 08TP C8 Silica Spheroidal,, 7, 0, 0, 00Å 60 0m /g % Polymeric Yes L7 0µm TP C Silica Spheroidal,, 7, 0, 0, 00Å 60 0m /g % Polymeric Yes L6 0µm 8TP C8 Silica Spheroidal,, 7, 0, 0, 00Å 60 0m /g 8% Polymeric Yes L 0µm 9TP Di-Phe Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g % Polymeric Yes 8TP C8 Silica Spheroidal,, 7, 0, 0, 00Å 60 0m /g % Monomeric Yes L 0µm Everest 8EV C8 Silica Spherical, 0, 0, 0µm 00Å 70 0m /g 6% Monomeric Yes L Vydac MS 08MS C8 Silica Spheroidal µm 00Å 70m /g % Polymeric Yes L7 MS C Silica Spheroidal µm 00Å 70 0m /g % Polymeric Yes L6 8MS C8 Silica Spheroidal,, 0, 0 µm 00Å 60 0m /g 8% Polymeric Yes L 8MS C8 Silica Spheroidal µm 00Å 70m /g % Monomeric Yes L 9MS Di-Phe Silica Spheroidal µm 00Å 70m /g % Polymeric Yes *Product information is available at. Proprietary. 0

8 Grace HPLC Hardware Formats Grace Davison Discovery Sciences provides hardware formats to maximize the performance of the phases in our column families for all applications. Choose a hardware format based on your method requirements for speed, sensitivity, and resolution. Ultra High-Pressure Hardware Analytical Hardware Ideally suited for ultra high-pressure systems, this ultralow volume hardware is pressure rated to 8,000psig, and packed with.µm media to maximize speed and efficiency. Lengths 0, 0, 0, 00mm Expedite MS Hardware Inner Pressure Wetted Diameters Limit Surfaces.0,.0mm 8,000psig 6 Stainless Steel High-speed, low-volume columns for rapid resolution and high-throughput LC/MS applications mm i.d. columns for standard HPLC instrumentation, the most commonly used. Analytical columns have industry standard port configurations. Some column families are also available with port configurations for Waters endfittings. Lengths 0, 0, 00, 0, 0, 00mm Metal-Free Hardware Inner Pressure Wetted Diameters Limit Surfaces.6mm 0,000psig 6 Stainless Steel 7 hplc columns introduction Lengths 0, 0mm Rocket Hardware Inner Pressure Wetted Diameters Limit Surfaces.,.6mm 0,000psig 6 Stainless Steel High-speed, high-resolution columns for high-throughput analysis. Large 7mm i.d. balances column volume with system volume to deliver excellent peak shapes on conventional HPLC instrumentation. Large i.d. also allows fast mobile-phase flow rates which minimizes peak broadening. Lengths, mm Solvent-Reducer and Microbore Hardware Smaller diameter columns reduce solvent consumption and increase sensitivity when compared to standard.6mm i.d. columns. Use with standard HPLC instrumentation or with MS and ELS detectors. Lengths 0, 00, 0, 0, 0, 00mm Inner Pressure Wetted Diameters Limit Surfaces 7mm 0,000psig 6 Stainless Steel Inner Diameters Pressure Limit Wetted Surfaces.0,.,.0mm 0,000psig 6 Stainless Steel Mechanically strong, metal-free columns offer biocompatibility, and chemical resistance that ion chromatography and biotechnology applications demand. Lengths 0, 00, 0, 0, 00mm Capillary Guards Inner Pressure Diameters Limit.,.6mm 000psig PEEK All-Guard Guard Cartridges Wetted Surfaces Capillary guards offer zero dead-volume with finger-tight connections to maintain column performance. Use them to protective your capillary investment, as enrichment columns, or as short analytical columns. Lengths, 0, 0mm Inner Diameters Pressure Limit Wetted Surfaces 0., 0.mm 000psig 6 Stainless Steel and fused silica Guard system with reusable holder and disposable guard columns to protect analytical columns Capillary Hardware Lengths 7.mm Inner Pressure Wetted Diameters Limit Surfaces.,.0,.6mm 000psig 6 Stainless Steel 7 For LC/MS and other high-sensitivity and sample-limited applications. Lengths 0, 00, 0, 0, 00mm Inner Diameters 0.07, 0.0, 0., 0., 0.8mm Pressure Limit 000psig Wetted Surfaces 6 Stainless Steel and fused silica more info Looking for preparative HPLC hardware? See page 0.

9 hplc columns introduction High Throughput HPLC Introduction The Benefits of Grace Small Particles and Formats Today s laboratories are under greater demand to analyze more samples in less time. To help meet this demand, major product advances have been made to decrease analysis time and increase HPLC throughput. HPLC system advances have greatly reduced the influence of extra column volumes and have extended the range of flow rates and pressure capabilities. In response to this, Grace now offers a wide range of column chemistries and formats appropriate for all types of high throughput systems. The use of smaller particles offers two main improvements to the chromatographic separation increased resolution and speed. Resolution is directly proportional to the square root of column efficiency, therefore the higher the efficiency, the narrower the peaks and the greater the resolution between them. Increased speed comes from higher mobile phase flow rates that can be used without loss in efficiency, and higher flow rates mean faster analysis times. Understanding the type of system currently in use for high throughput separations and then matching the right column configuration is critical to achieving the best high throughput separation. Here we outline the three types of systems currently in use today for high throughput and the recommended HPLC column format for use with that system. System Type Ultra High-Pressure LC System (>0,000psig pressure limitation) Examples: Agilent 00, Waters Acquity, Thermo Accela, Jasco XLc. Speed from Ultra High-Pressure Systems: Representing the latest in LC instrumentation technology, ultra high-pressure systems theoretically have the potential to deliver the fastest separations. They have minimal system volume and offer a pressure limit upwards of,000psig. This allows the use of columns with sub µm particles and 7 times traditional flow rates. Sub µm particles extend the working range of acceptable mobile phase linear velocities without sacrificing efficiency. Therefore, you can push flow rates and still get equal or better performance. Suggested Column Format: VisionHT columns are designed for microbore and ultra high-pressure LC systems that have small system volumes to limit peak broadening from sample diffusion. VisionHT columns are packed with highly efficient.µm phases that yield taller peaks and increase sensitivity. The column hardware incorporates a low dead volume design to minimize sample diffusion, and maintain peak integrity and efficiency.,000psig high-pressure stability allows fast flow rates, decreasing run times 0 fold. System Type Recommended Column Format: Length: i.d.: VisionHT 0, 0, 0, 00mm.0 and.0mm tech tip Converting Methods From Traditional Column Formats to High Throughput Columns 7 Convert Between Standard HPLC and VisionHT Columns Flow Rate Standard HPLC (.6mm).0X VisionHT (.0mm).X When adjusting between standard LC conditions to VisionHT columns convert flow rates accordingly and then increase flow rate for faster analysis. Convert Between Standard HPLC Columns and Rocket Columns Flow Rate Standard Analytical (.6mm).0X Rocket Column (7.0mm).X mau VisionHT Columns for Ultra High-Pressure LC Sulfadiazine. Sulfathiazole. Sulfamerazine. Sulfamethazine. Sulfamethoxazole 6. Sulfaisoxazole 7. Sulfaquinoxilone 8. Sulfamethoxine Use this conversion of flow rate to transfer methods between Rocket column or VisionHT columns. Backpressure on standard LC systems should be considered. 0 Min. System Agilent 00 Mobile Phase: A: 0.% Formic Acid B: Methanol Detector: UV at 80nm Temperature: Ambient more info See pages 7 for more information on VisionHT columns.

10 System Type Low Volume, High Throughput (HTP) LC System (<0,000psig Pressure Limitation) Examples: Shimadzu Prominence UFLC, Hitachi Ultra, LC Packings UltiMate, Surveyor Plus Speed from Low-Volume HTP LC Systems: These systems concentrate on reducing the cycle time between injections thus allowing more injections per time frame. Typically they employ high-speed gradient pumps, fast autosamplers and a quick detector sampling rates. They are not pressure rated to the extremes of ultra high-pressure systems, but the low system volume allows for short columns of narrow i.d. Suggested Column Format: Expedite and VisionHT columns are designed for microbore high throughput LC systems that have small system volumes. These formats incorporate a low dead volume design so that sample bands do not diffuse within the column hardware, maintaining peak integrity and efficiency. Expedite columns are packed with highly efficient.µm or µm in very short column lengths to minimize backpressure and reduce analysis times. Though not as fast as ultra highpressure LC systems these systems and columns balance both speed and backpressure. System Type Recommended Column Formats: Length: i.d.: 6 7 Expedite and VisionHT 0, 0, 0, 0mm,,.6mm 6 Expedite Columns for Low Volume High Throughput LC 076. Ribonuclease (7ng). Bovine Insulin (7ng). Cytochrome C (7ng). Lysozyme (ng). Bovine Serum Albumin (7ng) 6. Carbonic Anhydrase (7ng) 7. Ovalbumin (7ng) System Type Traditional LC System (<000psi Pressure Limitation) Examples: Agilent 00, Waters Alliance, Thermo Surveyor, Dionex Ultimate, Shimadzu Prominence, Hitachi LaChrom Speed from Traditional LC Systems: To get speed from a traditional LC system, you need to consider the 000psig pressure limitation, and the typically large ml system volume. High throughput columns on this system need to deliver a highly efficient separation typically achieved with a small particle packing ( µm), but without the high backpressures. The large system volume also needs to be balanced with an equally large column volume or the separation will be plagued with extra-column effects. Suggested Column Format: Rocket columns provide low backpressure and fast analysis times while preserving column efficiency. They are available with both.µm and µm packing materials for use on standard HPLC systems with backpressure limits less than 000psig. The 7mm i.d. allows faster flow rate that sweep the extra system volume faster and reduce peak broadening. This larger diameter also means a larger column volume to system volume ratio to minimize the efficiency loss from extra system volume. This benefit is more pronounced over. and mm i.d. columns that have a smaller ratio than.6mm i.d. columns and require much lower flow rates for acceptable backpressures. Low flow rates allow more time for sample diffusion within the standard HPLC s system volume to further degrade the column s efficiency. System Type Recommended Column Format: Length: i.d.: Packings: Rocket or mm 7mm Any.µm or µm media Rocket Columns for High Throughput with Conventional LC. Xanthine. Theobromine. Theophylline. Caffeine. 8-Chlorotheophylline hplc columns introduction 0 Min. Column: ProZap C8,.µm,. x 0mm Expedite Mobile Phase: A: 0.% TFA in Water B: 0.08% TFA in Acetonitrile Flow Rate: 0.8mL/min Detector: UV at 80nm Injection Vol.: µl 0 Min. Column: Platinum C8-EPS 00Å, µm, 7 x mm Rocket Mobile Phase: 0.00M Sodium Acetate, ph.0:methanol (70:0) Flow Rate:.0mL/min Detector: UV at nm more info Expedite columns are available in most popular Alltech brand phases. See page for overview of phases and formats offered. more info Rocket columns are available in most popular Alltech brand phases. See page for overview of phases and formats offered.

11 analytical hplc Grace VisionHT Ultra High-Pressure Columns See Separations with Speed and Clarity Ultra-fast separations with superior efficiency, sensitivity, and resolution Exceptional stability for long column lifetimes Comprehensive sub μm stationary phase offering,000psig pressure rating compatible with all ultra high-pressure LC systems Grace VisionHT columns offer a new level of performance in HPLC. The powerful combination of high-strength.µm media with ultra-low volume hardware, delivers new clarity to your separations and maintains exceptional column lifetime. Complex samples resolve 9% faster with x greater sensitivity when compared to traditional. x 0mm, µm columns. And with a wide variety of phases available, the possibilities are endless. 00 mau mau Min. Increase Speed by 9% and Gain x Better Sensitivity Column: VisionHT C8,.µm,.0 x 0mm Flow Rate: 0.8mL/min 00 mau Min. Column: Traditional C8, µm,. x 0mm Flow Rate: 0.mL/min 00 0 Impurities not seen on traditional µm columns Min.. Thiourea. Toluene. Propylbenzene. Butylbenzene System: Agilent 00 Mobile Phase: (:) 0mM Potassium Phosphate, ph.:acetonitrile Detector: UV at nm Temperature: Ambient Efficiency and Retention Times Remain Constant after Exposure to,000psig and 000 Injections Plates/Meter (000) Column Stability Comparison, 0 x mm VisionHT Column VisionHT Column VisionHT Column VisionHT Column Column Volumes VisionHT Column VisionHT Column 6 VisionHT Column 7 Competitor Sub µm Column Competitor Sub µm Column Uracil. Pyridine. Phenol. Methyl Alanine. -Butyl Benzoic Acid 6. Toluene Initial Min. FINAL (After 000 injections) Min. Column:.0 x 0mm Mobile Phase: 0mM KHPO, ph 7.0:CHCN (60:0) Flow Rate:.0mL/min High-pressure competitor sub µm column lost efficiency at routine,000psig pressures. VisionHT columns remain stable. Before and after chromatograms show a constant level of performance after 000 injections.

12 Grace VisionHT Ultra High-Pressure Columns Comprehensive High Purity.µm Selectivity Options Sub µm particles deliver efficiency and speed, but critical to success is having the right stationary phase selectivity. Six VisionHT high purity phases are available, each with unique separation benefits. C8-HL, with maximum bonded phase coverage, is ideal for complex hydrophobic samples. Use C8-B for basic compounds at neutral ph, often a requirement for mass spec work in the pharmaceutical industry. Reserve the C8 and C8-P for fastest analysis times. Both offer increased polar interactions to make neutral, non-polar compounds elute faster and retain polar compounds longer. The HILIC and Silica packings are normal phases that typically use near exclusive organic mobile phases; an advantage when seeking highest mass spec sensitivity. VisionHT phases have exceptionally rigid silica structure that withstands routine use of,000psig pressure often required in this new technology. Reproducible silica synthesis, bonding and column packing guarantee low bleed and excellent column-to-column consistency. VisionHT Phase Specifications Base Material Spherical Particle Carbon Pore Surface Size Load Size Area Endcapped.µm 0% 0Å 0m /g Yes 0 Fully bonded silica, ultra ph Range* Feature Benefits Packing C8-HL High capacity for hydrophobic compounds. Good Silica high purity silica peak symmetry. C8-B Spherical.µm % 0Å 0m /g Proprietary 0 High purity silica and Improved performance of basic compounds at Silica unique endcapping. neutral ph. Better sensitivity and peak shape by mass spec for basic compounds, without the need for acidified mobile phases. C8 Spherical.µm 6% 00Å 00m /g Yes 0 Moderate silica exposure Classic reversed-phase selectivity. Reduced Silica bonding is optimized for speed and sensitivity. C8-P Spherical.µm % 00Å 00m /g No 0 High silica exposure, low Unique polar selectivity. Low carbon load gives Silica carbon load fastest reversed-phase elution times while retaining HILIC Silica Spherical Silica Spherical Silica *Choice of buffer is critical at ph >8..µm NA 0Å 0m /g No 8 Polar phase with shorter equilibration times. Shipped in ACN/Water..µm NA 0Å 0m /g No 8 Traditional normal phase for use in 00% organic mobile phases polar compounds longer. Peak reversal compared to reversed-phase. Ideal for very polar compounds with high organic mobile phases for improved sensitivity by MS. For isomeric separation of non-aqueous compatible compounds by absorption chromatography k' 0 VisionHT Reversed-Phase Selectivity k' Phenol k' Methyl Alanine k' Butyl Benzoic Acid k' Toluene C8-P C8 C8-B VisionHT Phase C8-HL technical assistance Contact Tech Support: Phone: (North America) contact.alltech@grace.com Online: Separate Highly Basic Components at Neutral ph with Vision HT C8-B. Phenylephrine. Diphenhydramine. Amitriptyline 0 Min. Column: VisionHT C8-B,.µm,.0 x 0mm Mobile Phase: 0mM Ammonium Formate ph 7:Methanol (0:80) Column Temp: 0 C Flow Rate: 0.mL/min Detector: UV at 0nm

13 Grace VisionHT Ultra High-Pressure Columns Unique Column Hardware Advantages You can erode the benefits of sub µm media if packed in an inefficient column design. VisionHT hardware uses thin screens, instead of traditional frits, to retain media and minimize dead volume. A unique insert combination seals endfittings leak-free to 8,000psig. Various dimensions offer even more flexibility over analysis speed and resolution. VisionHT Hardware Specifications Dead volume: <nl Wetted materials: 6 Stainless Steel, PTFE Port Geometry: 0, for Industry Standard & Waters connection Pressure rating, hardware alone: 8,000psig (0 bar) Pressure rating, packed column:,000psig (80 bar) i.d.:.0,.0mm Length: 0, 0, 0, 00mm The Best Value for Ultra High-Pressure Method Development UltraMD Kits! Whether developing a new method, or improving an existing analysis, the VisionHT UltraMD kits can help optimize your separation. Get four reversed-phase columns with varying selectivity for the price of three, and develop your fast LC method fast! 76 VisionHT UltraMD Kits Phases Dimensions Part No. UltraMD Kit C8, C8-P, C8-HL, C8-B x 00mm 69 UltraMD Kit C8, C8-P, C8-HL, C8-B x 0mm 69 UltraMD Kit C8, C8-P, C8-HL, C8-B x 0mm 69 7 VisionHT Reversed-Phase Comparison C8-P C8-B 0 6 Min. C8 C8-HL. Uracil. Pyridine. Phenol. Methyl Alanine. -Butyl Benzoic Acid 6. Toluene Column:.0 x 0mm Mobile Phase: 0mM KHPO, ph 7.0:CHCN (60:0) Flow Rate:.0mL/min 6

14 Grace VisionHT Ultra High-Pressure Columns Protect Your UHPLC Column Investment The smaller diameters and finer porosity frits required in UHPLC creates a higher likelihood of column and system blockage. Trace contaminates not normally problematic for traditional LC, may now pose backpressure issues. VisionHT guards protect your column to minimize down time and reduce cost. Two guard formats available: Integral and stand-alone. Couple the zero-dead volume integral guard directly to any VisionHT column, and realize absolutely no loss in efficiency. Use the stand-alone version to protect other manufacturers UHPLC columns, or as an on-line enrichment device. Prevent Column Blockage with Integral Guards with guard without guard Min. No loss in efficiency. VisionHT Integral Guard System N=7K/m Replace existing endfitting with the integral guard endfitting. Change guard cartridges as necessary. 76 VisionHT Ultra High-Pressure Columns Packing Format i.d. x Length Part No. C8-HL,.µm Ultra High-Pressure x 0mm 0 Ultra High-Pressure x 0mm Ultra High-Pressure x 0mm Ultra High-Pressure x 00mm Ultra High-Pressure x 0mm Ultra High-Pressure x 0mm Ultra High-Pressure x 0mm 6 Ultra High-Pressure x 00mm 7 C8-B,.µm Ultra High-Pressure x 0mm 90 Ultra High-Pressure x 0mm 90 Ultra High-Pressure x 00mm 90 Ultra High-Pressure x 0mm 90 Ultra High-Pressure x 0mm 906 Ultra High-Pressure x 0mm 907 Ultra High-Pressure x 0mm 908 Ultra High-Pressure x 00mm 909 C8,.µm Ultra High-Pressure x 0mm 9 Ultra High-Pressure x 0mm 99 Ultra High-Pressure x 0mm 960 Ultra High-Pressure x 00mm 9607 Ultra High-Pressure x 0mm 97 Ultra High-Pressure x 0mm 9600 Ultra High-Pressure x 0mm 960 Ultra High-Pressure x 00mm 9609 C8-P,.µm Ultra High-Pressure x 0mm 96 Ultra High-Pressure x 0mm 960 Ultra High-Pressure x 0mm 960 Ultra High-Pressure x 00mm 9608 Ultra High-Pressure x 0mm 98 Ultra High-Pressure x 0mm 960 Ultra High-Pressure x 0mm 9606 Ultra High-Pressure x 00mm 960 HILIC,.µm Ultra High-Pressure x 0mm 90 Ultra High-Pressure x 0mm 9 Ultra High-Pressure x 0mm 9 Ultra High-Pressure x 00mm 9 Ultra High-Pressure x 0mm 96 Ultra High-Pressure x 0mm 97 Ultra High-Pressure x 0mm 99 Ultra High-Pressure x 00mm 90 SI,.µm Ultra High-Pressure x 0mm 9 Ultra High-Pressure x 00mm 9 Ultra High-Pressure x 0mm 9 Ultra High-Pressure x 00mm 9 Stand-Alone Guard System In-line guard system expands applications beyond column protection. Use as a sample enrichment device or to protect any uhplc commercially available column. 77 VisionHT Guard Cartridges Packing Format i.d. x Length Qty. Part No. C8HL,.µm Ultra High-Pressure x mm /pk 9 Ultra High-Pressure x mm /pk 8 C8 B,.µm Ultra High-Pressure x mm /pk 9 Ultra High-Pressure x mm /pk 9 C8,.µm Ultra High-Pressure x mm /pk 90 Ultra High-Pressure x mm /pk 9 C8-P,.µm Ultra High-Pressure x mm /pk 9 Ultra High-Pressure x mm /pk 9 HILIC,.µm Ultra High-Pressure x mm /pk 9 Ultra High-Pressure x mm /pk 9 SI,.µm Ultra High-Pressure x mm /pk 97 Ultra High-Pressure x mm /pk 96 Integral Guard Column Holder For VisionHT 8 Stand-Alone Guard Holder 80 7

15 Alltech Alltima HP Introduction High-Stability, High-Purity, High-Performance, Low-Bleed Columns for Demanding Applications Better Peak Symmetry high-purity silica eliminates peak tailing problems Long Column Life exceptional column stability minimizes downtime and reduces cost Ideal for Microbore or Critical Analysis low to no detectable column bleed Variety of Phases and Formats optimizes retention, resolution, and analysis time Alltima HP combines our best phase chemistries with high-purity silica. The result is one product family with selectivity and performance needed to overcome your most challenging separation needs. Alltech Alltima HP Phase Specifications Pore Size Surface Area Carbon Load Endcapped? Phase Description Base Particle Particle Size Phase USP C8 For classic reversed-phase applications Silica Spherical, µm 90Å 00 m /g % Monomeric Yes L C8 EPS Reversed-phase C8 with extended polar selectivity Silica Spherical, µm 90Å 00 m /g % Monomeric Yes L C8 HiLoad Higher carbon load for stronger retention Silica Spherical, µm 00Å 0 m /g % Monomeric Yes L C8 AQ 00% water wettable Silica Spherical, µm 00Å 0 m /g 0% Monomeric Yes L C8 Amide Low bleed polar-embedded phase compatible Silica Spherical, µm 90Å 00 m /g % Monomeric Yes L with microbore C8 For reversed-phase applications where C8 is Silica Spherical, µm 90Å 00 m /g 8% Monomeric Yes L7 too retentive Cyano Stable, long-life cyano phase Silica Spherical, µm 90Å 00 m /g % Monomeric Yes L0 Silica For general purpose normal-phase applications Silica Spherical, µm 00Å 0 m /g No L HILIC Hydrophillic Interaction Chromatography for highly polar analytes Silica Spherical.,, µm 0Å 0 m /g L Basic Compounds at Neutral ph Peak USP Tailing*. Diphenhydramine.9. Nortriptyline.. Amitriptyline.7 *Tailing Factor was determined in accordance with USP method. Column: Alltima HP C8, µm,.6 x 0mm Mobile Phase: mm Potassium Phosphate, ph 7.0:Methanol (:8) Flow Rate:.0mL/min Detector: UV at nm Min. The Alltima HP phases demonstrate excellent peak symmetry of highly basic analytes at ph 7.0. Basic Compound Stability at ph and ph0 Final Acid Final Base Retention Times. Uracil Phenol N,N-Diethyl-m-Toluamide Toluene Column: Alltima HP C8, µm,.6 x 0mm Mobile Phase: Acetonitrile:Water (8:) Flow Rate:.0mL/min Detector: UV at nm Final Acid: washed with,000 column volumes of Acetonitrile:Dilute Sulfuric Acid, ph.0 (0:0) at 60 C. Final Base: washed with,000 column volumes of Acetonitrile:Ammonium Hydroxide, ph 0 (0:0) at 0 C. Initial 0 Min. The Alltima HP phases demonstrate exceptional stability from ph.0 to 0. 8

16 Alltech Alltima HP Introduction The Alltima HP Family Demonstrates Low Bleed Alltima HP columns are among the lowest-bleed columns available, making them ideal for microbore applications. Our C8 Amide polar-embedded phase has significantly lower column bleed than competitive amide columns. Relative Abundance Level (X0 ) Column Bleed by ESI-MS Alltima HP vs. Competitive Columns Alltech Alltima HP Competitors No Column Alltima HP Silica C8 Alltima HP Alltima HP C8 HiLoad C8 MS C8 MS Alltima HP C8 Amide C8 Amide C8 Amide Alltima HP Method Development Kits best value Three Different Kits with Different Selectivities Whether you are developing a new method, or just not satisfied with an existing separation, the Alltima HP family can help optimize your separation. Your best value for method development purchase a kit and receive three columns for the price of two Alltima HP Method Development Kits Description Includes: i.d. x Length Part No. Kit C8 Mix, µm Expedite MS Kit C8, C8 HiLoad, C8 Amide (8767, 8769, 8778). x 0mm 878 Analytical Kit C8, C8 HiLoad, C8 Amide (87668, 87686, 877).6 x 0mm 8780 Rocket Kit C8, C8 HiLoad, C8 Amide (8767, 87690, 8776) 7 x mm 878 Kit Alternate Mix, µm Expedite MS Kit C8 HiLoad, C8, CN (8769, 8776, 8776). x 0mm 8786 Analytical Kit C8 HiLoad, C8, CN (87686, 8770, 87760).6 x 0mm Rocket Kit C8 HiLoad, C8, CN (87690, 877, 8776) 7 x mm 8786 Kit Polar Mix, µm Expedite MS Kit C8 EPS, C8 Amide, CN (8770, 8778, 8776). x 0mm 8787 Analytical Kit C8 EPS, C8 Amide, CN (8770, 877, 8778).6 x 0mm Rocket Kit C8 EPS, C8 Amide, CN (87708, 8776, 8776) 7 x mm 8787 tech tip Want high throughput on a conventional (non-ultra high-pressure) system? Choose the Rocket column format no modifications necessary to your existing HPLC system. Choose Expedite columns for low volume and microbore systems. See page for more explanation of the Rocket and Expedite formats. 9

17 0 Alltech Alltima HP Reversed-Phase Columns Selectivity Options in One Product Family C8: Classic reversed-phase retention and selectivity C8 AQ: 00% water wettable and high carbon loading for greater mobile phase range EPS C8: Unique selectivity that succeeds when traditional reversed-phase columns fail Greater retention and enhanced peak symmetry for polar compounds C8 HiLoad: Superior retention and loadability for resolution of complex samples C8 Amide: The first polar-embedded packing ideal for microbore Excellent peak shape without phase bleed Ideal for basic compounds in neutral to alkaline ph C8: Less hydrophobic retention than C8 Alltima HP Selectivity Alltima HP Packing Acid, Base, and Neutral Test Mix Min. Column:.6 x 0mm Mobile Phase: 0mM KHPO, ph.0:chcn (0:60) Column Temp: Ambient ( C) Flow Rate:.0mL/min Injection Vol.: µl Detector: UV at nm 6 6. Uracil. Pyridine. Phenol. N,N-diethyl-m-toluamide. -Butyl Benzoic Acid 6. Toluene C8 EPS C8 C8 HiLoad Alltima HP HPLC Columns Packing Format i.d. x Length Part No. C8, µm* Capillary 0.0 X 0mm Capillary 0.0 X00mm Capillary 0.0 X0mm Capillary 0.00 X 0mm 6 Capillary 0.00 X00mm 7 Capillary 0.00 X0mm 8 Expedite MS. x 0mm 8767 Expedite MS. x 0mm 8767 Microbore. x 0mm 870 Microbore. x 00mm Microbore. x 0mm Solvent-Reducer.0 x 0mm 8760 Expedite MS.6 x 0mm 8767 Expedite MS.6 x 0mm Analytical.6 x 0mm 8786 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 8767 Rocket 7 x mm 8767 C8, µm Microbore. x 0mm 8768 Microbore. x 0mm 8768 Solvent-Reducer.0 x 0mm 8760 Solvent-Reducer.0 x 0mm 8760 Analytical.6 x 0mm Analytical.6 x 0mm C8 AQ, µm* Capillary 0.0 x 0mm 60 Capillary 0.0 x 00mm 6 Capillary 0.0 x 0mm 6 Capillary 0.00 x 0mm 6 Capillary 0.00 x 00mm 6 Capillary 0.00 x 0mm 6 Expedite MS. x 0mm 878 Expedite MS. x 0mm 878 Microbore. x 00mm Microbore. x 0mm 8780 Expedite MS.6 x 0mm 878 Expedite MS.6 x 0mm 8786 Analytical.6 x 0mm 878 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 878 Rocket 7 x mm 878 C8 AQ, µm Microbore. x 0mm 878 Microbore. x 0mm 878 Analytical.6 x 0mm 8789 Analytical.6 x 0mm 8780 C8 EPS, µm Expedite MS. x 0mm Expedite MS. x 0mm 8770 Microbore. x 0mm 8708 Microbore. x 00mm 8770 Microbore. x 0mm Solvent-Reducer.0 x 0mm 8760 Expedite MS.6 x 0mm 877 Expedite MS.6 x 0mm 877 Analytical.6 x 0mm 878 Analytical.6 x 00mm 8770 Analytical.6 x 0mm 8770 Rocket 7 x mm Rocket 7 x mm C8 EPS, µm Microbore. x 0mm 8777 Microbore. x 0mm 8778 Solvent-Reducer.0 x 0mm 8760 Solvent-Reducer.0 x 0mm Analytical.6 x 0mm 877 Analytical.6 x 0mm 8776 *Other particle sizes and dimensions are available.

18 Alltech Alltima HP Reversed-Phase Columns Alltima HP HPLC Columns (continued) Packing Format i.d. x Length Part No. C8 Hi-Load, Capillary 0.0 x 0mm 90 µm* Capillary 0.00 x 0mm 9 Expedite MS. x 0mm 8769 Expedite MS. x 0mm 8769 Microbore. x 0mm 8706 Microbore. x 00mm Microbore. x 0mm Solvent-Reducer.0 x 0mm 8760 Expedite MS.6 x 0mm 8769 Expedite MS.6 x 0mm 8769 Analytical.6 x 0mm 8787 Analytical.6 x 00mm 8768 Analytical.6 x 0mm Rocket 7 x mm Rocket 7 x mm C8 Hi-Load, Microbore. x 0mm µm Microbore. x 0mm Solvent-Reducer.0 x 0mm 876 Solvent-Reducer.0 x 0mm 876 Analytical.6 x 0mm Analytical.6 x 0mm C8 Amide, Capillary 0.0 x 0mm µm* Capillary 0.00 x 0mm 8 Expedite MS. x 0mm 8777 Expedite MS. x 0mm 8778 Microbore. x 0mm 870 Microbore. x 00mm 877 Microbore. x 0mm 877 Solvent-Reducer.0 x 0mm Expedite MS.6 x 0mm 8779 Expedite MS.6 x 0mm 8770 Analytical.6 x 0mm 8789 Analytical.6 x 00mm 877 Analytical.6 x 0mm 877 Rocket 7 x mm 877 Rocket 7 x mm 8776 C8 Amide, Microbore. x 0mm 877 µm Microbore. x 0mm 8776 Solvent-Reducer.0 x 0mm Solvent-Reducer.0 x 0mm Analytical.6 x 0mm 877 Analytical.6 x 0mm 877 C8, µm* Capillary 0.0 x 0mm 79 Capillary 0.0 x 00mm 80 Capillary 0.0 x 0mm 8 Capillary 0.00 x 0mm 8 Capillary 0.00 x 00mm 8 Capillary 0.00 x 0mm 8 Expedite MS. x 0mm 877 Expedite MS. x 0mm 8776 Microbore. x 0mm 87 Microbore. x 00mm 877 Microbore. x 0mm 877 Solvent-Reducer.0 x 0mm 876 Expedite MS.6 x 0mm 8777 Expedite MS.6 x 0mm 8778 Analytical.6 x 0mm 8780 Analytical.6 x 00mm 8779 Analytical.6 x 0mm 8770 Rocket 7 x mm 877 Rocket 7 x mm 877 C8, µm Microbore. x 0mm 877 Microbore. x 0mm 877 Solvent-Reducer.0 x 0mm 876 Solvent-Reducer.0 x 0mm 876 Analytical.6 x 0mm 877 Analytical.6 x 0mm 877 *Other particle sizes and dimensions are also available. Alltima HP-Guard Cartridges Packing i.d. x Length Qty. Part No. C8 Capillary Guard, µm** 0.0 x 0mm ea x 0mm ea 79 C8 All-Guard, µm*. x 7.mm x 7.mm x 7.mm 8768 C8 AQ Capillary Guard, µm** 0.0 x 0mm ea x 0mm ea 69 C8 AQ All-Guard, µm*. x 7.mm x 7.mm 878 C8 EPS All-Guard, µm*. x 7.mm x 7.mm x 7.mm 8770 C8 Hi-Load Capillary 0.0 x 0mm ea 88 Guard, µm** 0.00 x 0mm ea 89 C8 Hi-Load All-Guard, µm*. x 7.mm x 7.mm x 7.mm 8770 C8 Amide Capillary 0.0 x 0mm ea 98 Guard, µm** 0.00 x 0mm ea 99 C8 Amide All-Guard, µm*. x 7.mm x 7.mm x 7.mm 8778 C8 Capillary Guard, µm** 0.0 x 0mm ea x 0mm ea 68 C8 All-Guard, µm*. x 7.mm x 7.mm x 7.mm 8776 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) Capillary Guard Cartridge Holder Guard Holder for 0.00mm and 0.0mm Guards ea GR-70E Guard Holder for 0.00mm and 0.00mm Guards ea GR-70A *All-Guard holder required. **Other particle sizes and dimensions are available. tech tip Which guard format should I choose? Ideally, the guard should have the same or slightly smaller i.d. as the HPLC column. Efficiency will suffer, if the guard i.d. is larger than that of the column. technical assistance Contact Tech Support: Phone: (North America) contact.alltech@grace.com Online:

19 Alltech Alltima HP HILIC Columns Hydrophilic Interaction (HILIC) Columns for Highly Polar Basic Analytes Superior retention of highly polar compounds Increased sensitivity and lower detection limits with microbore Available in.µm particle size Hydrophilic Interaction Chromatography is ideal for separating and retaining very polar compounds that may not retain on traditional reversed-phase packings. Unlike reversed-phase columns, HILIC columns retain highly polar compounds with only small amounts of water in the mobile phase. These more volatile mobile phases increase sensitivity with microbore applications. Basic Drugs Reverse-Phase C8 Columns Min. Column: Mobile Phase: Flow Rate: Detector: Mixed Analyte HILIC. Toluene. Hydantoin. Uracil. Caffeine. Allantoin Alltima HP HILIC, μm,.6 x 0mm (Part No. 866) Acetonitrile:0.0M Ammonium Formate, ph.0 (9:).0mL/min UV at 0nm 06 Alternate selectivity compared to reversed-phase columns allows more separation possibilities. Alltima HP HILIC Columns Min.. Albuterol. Bamethan Column: Prevail C8 Column, μm,.6 x 0mm (Part No. 9908) Alltima HP HILIC Column, μm,.6 x 0mm, (Part No. 866) Mobile Phase: 0.M Ammonium Formate, ph.0:chcn (88:) Acetonitrile:0.M Ammonium Formate, ph.0 (8:) Flow Rate:.0mL/min Detector: UV at 0nm Use Alltima HP HILIC's alternate selectivity for greater separation possibilities. tech tip Want high throughput on a conventional (non-ultra high-pressure) system? Choose the Rocket column format no modifications necessary to your existing HPLC system. Choose Expedite columns for low volume and microbore systems. See page for more explanation of the Rocket and Expedite formats. Rocket Columns 690 Expedite Columns 6 Alltima HP Hilic HPLC Columns Packing Format i.d. x Length Part No. HILIC,.µm Expedite MS. x 0mm 867 Expedite MS.6 x 0mm 867 Rocket 7 x mm 8667 Rocket 7 x mm 8668 HILIC, µm* Capillary 0.0 x 0mm Capillary 0.0 x 00mm 6 Capillary 0.0 x 0mm 7 Capillary 0.00 x 0mm 8 Capillary 0.00 x 00mm 9 Capillary 0.00 x 0mm 0 Expedite MS. x 0mm 867 Expedite MS. x 0mm 867 Microbore. x 0mm 866 Microbore. x 0mm 866 Expedite MS.6 x 0mm 867 Expedite MS.6 x 0mm 8676 Analytical.6 x 0mm 8660 Analytical.6 x 0mm 866 Rocket 7 x mm 8669 Rocket 7 x mm 8670 HILIC, µm Microbore. x 0mm 866 Analytical.6 x 0mm 866 Analytical.6 x 0mm 8666 *Other particle sizes and dimensions are available. Alltima HP HILIC Guard Cartridges Packing i.d. x Length Qty. Part No. HILIC Capillary Guard, 0.0 x 0mm ea 68 µm** 0.00 x 0mm ea 69 HILIC All-Guard, µm*. x 7.mm x 7.mm 8680 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) Capillary Guard Cartridge Holder Guard Holder for 0.00mm and 0.0mm Guards ea GR-70E Guard Holder for 0.00mm and 0.00mm Guards ea GR-70A *All-Guard holder required. **Other particle sizes and dimensions are available.

20 Alltech Alltima HP Normal-Phase Columns Cyano: Superior stability More reproducible separations than silica for normal-phase applications Ideal for basic drug analysis Silica: Normal phase % Changes in Retention Alltima HP Cyano Much More Stable than Other Cyanos ph.0 ph 7. Alltima HP CN Brand A CN Brand B CN Brand C CN Brand D CN Brand E CN Brand F CN QC D Test: 70:0 HO/CHCN,.0mL/min, UV at nm, µl DMix, Column.6 x 0mm, C ph.0 Stability: Flushed with HSO ph.0:acetonitrile 0:0 at.0ml/min for days ph 7. Stability: Flushed with 0mM KHPO ph 7.:Acetonitrile 0:0 at.0ml/min for days Alltima HP HPLC Columns Packing Format i.d. x Length Part No. Cyano, µm Expedite MS. x 0mm 8776 Expedite MS. x 0mm 8776 Microbore. x 0mm 87 Microbore. x 00mm 8779 Microbore. x 0mm Solvent-Reducer.0 x 0mm 8766 Expedite MS.6 x 0mm 8776 Expedite MS.6 x 0mm Analytical.6 x 0mm 878 Analytical.6 x 00mm 8777 Analytical.6 x 0mm 8778 Rocket 7 x mm 8776 Rocket 7 x mm 8776 Cyano, µm Microbore. x 0mm 8778 Microbore. x 0mm 8778 Solvent-Reducer.0 x 0mm 8767 Solvent-Reducer.0 x 0mm 8768 Analytical.6 x 0mm Analytical.6 x 0mm Silica, µm Expedite MS. x 0mm 8779 Expedite MS. x 0mm 8779 Microbore. x 0mm 876 Microbore. x 00mm Microbore. x 0mm Solvent-Reducer.0 x 0mm 8769 Expedite MS.6 x 0mm 8779 Expedite MS.6 x 0mm 8779 Analytical.6 x 0mm 878 Analytical.6 x 00mm 8778 Analytical.6 x 0mm Rocket 7 x mm Rocket 7 x mm Deoxycorticosterone Acetate. Hydrocortisone Acetate. Cortisone Acetate. Cortisone Corticosteroids Min. Column: Alltima HP CN, μm,.6 x 0mm (Part No ) Mobile Phase: Hexane:Isopropanol (9.7:6.) Flow Rate:.0mL/min Column Temp: C Detector: UV at nm 6. Prednisone 6. Hydrocortisone 7. Prednisolone 7 09 Alltima HP HPLC Columns (continued) Packing Format i.d. x Length Part No. Silica, µm Microbore. x 0mm Microbore. x 0mm 8780 Solvent-Reducer.0 x 0mm 8760 Solvent-Reducer.0 x 0mm 876 Analytical.6 x 0mm Analytical.6 x 0mm Alltima HP All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Cyano, µm. x 7.mm x 7.mm x 7.mm 8778 Silica, µm. x 7.mm x 7.mm x 7.mm 8780 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. more applications To view our complete searchable chromatogram database visit /chromdb/

21 Alltech Prevail Introduction Stable from 00% Organic to 00% Aqueous Long column life in both highly aqueous and highly organic mobile phases Excellent retention and reproducibility of highly polar analytes in 00% aqueous mobile phases Strong retention of hydrophobic analytes in 00% organic mobile phases, no concerns of solubility, better sensitivity in microbore and ELSD applications No phase collapse Specialty phases for specific applications With one Prevail C8 column, retain polar compounds in aqueous mobile phases and hydrophobic compounds in organic mobile phases. Alltech Prevail Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Select Silica Spherical, µm 0Å 0m /g 7% Monomeric Yes L C8 Silica Spherical, µm 0Å 0m /g % Monomeric Yes L C8 Silica Spherical, µm 0Å 0m /g 8% Monomeric Yes L7 Phenyl Silica Spherical, µm 0Å 0m /g 7% Monomeric Yes L Cyano Silica Spherical, µm 0Å 0m /g Monomeric Yes L0 Amino (NH) Silica Spherical, µm 0Å 0m /g Monomeric No L8 Silica Silica Spherical, µm 0Å 0m /g L Organic Acid Silica Spherical, µm 0Å 0m /g Monomeric Yes Carbohydrate ES Polymer Spherical µm Switch Between 00% Aqueous and 00% Organic Mobile Phases on the Same Column Water-Soluble Vitamins (00% Aqueous) 96 Fat-Soluble Vitamins and Tocopherols (00% Organic) 90 Water-Soluble Vitamins (00% Aqueous) 96. Thiamine. Ascorbic Acid. Nicotinic Acid. Pyridoxine. Niacinamide 6. Pantothenic Acid. Vitamin A. δ-tocopherol. γ-tocopherol. α-tocopherol (Vitamin E). Vitamin D 6. Vitamin D 7. Vitamin K 6. Thiamine. Ascorbic Acid. Nicotinic Acid. Pyridoxine. Niacinamide 6. Pantothenic Acid 6 wash with intermediate 7 wash with intermediate Min Min Min. Column: Mobile Phase: Flow Rate: Detector: Prevail C8, µm,.6 x 0mm mm KHPO ph.0:chcn (97:).0mL/min UV at nm Column: Prevail C8, µm,.6 x 0mm Mobile Phase: CHCN:CHOH (7:) Flow Rate:.mL/min Detector: UV at 0nm Column: Mobile Phase: Flow Rate: Detector: Prevail C8, µm,.6 x 0mm mm KHPO ph.0: CHCN (97:).0mL/min UV at nm Figure Figure Figure Prevail Columns have highly stable bonded phases that let you use one column for multiple mobile phase conditions. Even switching between extremes such as near 00% aqueous (Figure ) to 00% organic conditions (Figure ), and back to near 00% aqueous (Figure ) on a routine basis is possible, provided that mobile phases are miscible.

22 Alltech Prevail Reversed-Phase Columns Prevail HPLC Columns Packing Format i.d. x Length Part No. Select C8, Microbore. x 0mm 9909 µm Microbore. x 00mm 99 Microbore. x 0mm 99 Solvent-Reducer.0 x 0mm 99 Analytical.6 x 00mm 990 Analytical.6 x 0mm 990 Rocket 7 x mm 990 Rocket 7 x mm 990 Select C8, Microbore. x 0mm 990 µm Microbore. x 0mm 99 Solvent-Reducer.0 x 0mm 996 Solvent-Reducer.0 x 0mm 997 Analytical.6 x 0mm 9900 Analytical.6 x 0mm 990 C8, µm Microbore 0µm x 0mm 76 Microbore 0µm x 0mm 76 Microbore 00µm x 0mm 8 Microbore 00µm x 0mm 8 Microbore.0 x 00mm 8 Microbore.0 x 0mm 8 Expedite MS. x 0mm 86 Expedite MS. x 0mm 87 Microbore. x 0mm 88 Microbore. x 00mm 87 Microbore. x 0mm 9900 Solvent-Reducer.0 x 0mm 99 Expedite MS.6 x 0mm 878 Expedite MS.6 x 0mm 80 Analytical.6 x 0mm 89 Analytical.6 x 00mm 990 Analytical.6 x 0mm 990 Rocket 7 x mm 9980 Rocket 7 x mm 9979 tech tip Want high throughput on a conventional (non-ultra high-pressure) system? Choose the Rocket column format no modifications necessary to your existing HPLC system. Choose Expedite columns for low volume and microbore systems. See page for more explanation of the Rocket and Expedite formats. Prevail HPLC Columns (continued) Packing Format i.d. x Length Part No. C8, µm Microbore 0µm x 0mm 79 Microbore 0µm x 0mm 79 Microbore 00µm x 0mm 8 Microbore 00µm x 0mm 86 Microbore. x 0mm 9906 Solvent-Reducer.0 x 0mm 990 Solvent-Reducer.0 x 0mm 99 Analytical.6 x 0mm 90 Analytical.6 x 0mm 9908 Analytical.6 x 0mm 990 C8, µm Microbore. x 0mm 99 Solvent-Reducer.0 x 0mm 99 Analytical.6 x 0mm 9 Analytical.6 x 00mm 99 Analytical.6 x 0mm 996 C8, µm Microbore. x 0mm 998 Solvent-Reducer.0 x 0mm 99 Solvent-Reducer.0 x 0mm 99 Analytical.6 x 0mm 99 Analytical.6 x 0mm 999 Phenyl, µm Microbore.0 x 00mm 89 Microbore.0 x 0mm 89 Expedite MS. x 0mm 87 Expedite MS. x 0mm 88 Microbore. x 0mm 89 Microbore. x 00mm 87 Microbore. x 0mm 99 Solvent-Reducer.0 x 0mm 998 Expedite MS.6 x 0mm 887 Expedite MS.6 x 0mm 8 Analytical.6 x 0mm 869 Analytical.6 x 00mm 99 Analytical.6 x 0mm 99 Rocket 7 x mm 998 Rocket 7 x mm 998 Phenyl, µm Microbore. x 0mm 997 Solvent-Reducer.0 x 0mm 996 Solvent-Reducer.0 x 0mm 997 Analytical.6 x 0mm 999 Analytical.6 x 0mm Prevail Bonded Phases are Stable in 00% Aqueous Conditions Prevail C8, µm,.6 x 0mm Competitor A C8, µm,.6 x 0mm Catecholamines. Norepinephrine. Epinephrine. -Hydroxydopamine Mobile Phase: mm KHPO, ph.7 Flow Rate:.0mL/min Detector: UV at 70nm After hrs. After 8 hrs. After 0 min Min. 0 6 Min.

23 Alltech Prevail Normal-Phase Columns Prevail HPLC Columns Packing Format i.d. x Length Part No. Cyano, µm Microbore. x 0mm 99 Solvent-Reducer.0 x 0mm 998 Analytical.6 x 0mm 9 Analytical.6 x 00mm 99 Analytical.6 x 0mm 997 Cyano, µm Microbore. x 0mm 999 Solvent-Reducer.0 x 0mm 999 Solvent-Reducer.0 x 0mm 990 Analytical.6 x 0mm 99 Analytical.6 x 0mm 99 Amino, µm Analytical.6 x 0mm 96 Analytical.6 x 00mm 997 Analytical.6 x 0mm 999 Amino, µm Analytical.6 x 0mm 996 Analytical.6 x 0mm 996 Silica, µm Microbore.0 x 00mm 8 Microbore.0 x 0mm 806 Expedite MS. x 0mm 8 Expedite MS. x 0mm 86 Microbore. x 0mm 868 Microbore. x 00mm 80 Microbore. x 0mm 9967 Solvent-Reducer.0 x 0mm 99 Expedite MS.6 x 0mm 88 Expedite MS.6 x 0mm 86 Analytical.6 x 0mm 8 Analytical.6 x 00mm 9969 Analytical.6 x 0mm 997 Rocket 7 x mm 998 Rocket 7 x mm 998 Prevail HPLC Columns (continued) Packing Format i.d. x Length Part No. Silica, µm Analytical. x 0mm 997 Solvent-Reducer.0 x 0mm 999 Solvent-Reducer.0 x 0mm 990 Analytical.6 x 0mm 997 Analytical.6 x 0mm 9977 Prevail All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Select C8, µm. x 7.mm x 7.mm x 7.mm 96 C8, µm. x 7.mm x 7.mm x 7.mm 9986 C8, µm. x 7.mm x 7.mm 99.6 x 7.mm 9987 Phenyl, µm. x 7.mm x 7.mm 99.6 x 7.mm 9988 Cyano, µm. x 7.mm 99.0 x 7.mm 99.6 x 7.mm 9989 Amino, µm.6 x 7.mm 9990 Silica, µm. x 7.mm 99.0 x 7.mm 99.6 x 7.mm 999 All-Guard Cartridge Holder ea 800 *All-Guard holder required. Other particle sizes available. Alltech Prevail Organic Acid Columns Get Unsurpassed Resolution for Common Organic Acids Acid stable for long column lifetimes Silica-based for maximum efficiency and high-resolution Short run times and high sample throughput at ambient temperature with Rocket format Lower cost than polymeric organic acid columns Prevail Organic Acid (OA) columns separate common organic acids with an unsurpassed combination of resolution, speed, sensitivity, and simplicity. A simple acidic phosphate buffer and a Prevail OA column at ambient temperature will separate short-chain organic acids in less than 6 minutes. Use ph to adjust column selectivity. Lowering the mobile phase ph progressively suppresses the ionization of the carboxylic acids, making them more hydrophobic. This gives you the ability to move these peaks relative to other peaks in the chromatogram, and it simplifies method development. 8 Figure - Organic Acids. Oxalic Acid. Tartaric Acid. Formic Acid 67. Malic Acid. Lactic Acid 9 6. Acetic Acid 0 7. Diglycollic Acid 8. Maleic Acid 9. Citric Acid Min. 0. Succinic Acid. Fumaric Acid Column: Prevail Organic Acid, µm,.6 x 0mm (Part No. 8860) Mobile Phase: mm KHPO, ph. Flow Rate:.mL/min Detector: UV at 0nm Separate short-chain organic acids in less than six minutes. 98 Prevail HPLC Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length OA, µm Microbore. x 00mm 8868 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 999 Rocket 7 x mm 07 OA, µm Analytical.6 x 0mm Analytical.6 x 0mm Prevail All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Organic Acid, µm.6 x 7.mm 969 All-Guard Cartridge Holder ea 800 *All-Guard holder required. Other particle sizes available. 6

24 Alltech Prevail Carbohydrate ES Columns Get Better Retention Times, Longer Column Life, and Improved Baselines Compared to Traditional Amino Carbohydrate Columns Longer column life: Hybrid phase has the ruggedness of silica with the stability of a polymer Quieter baselines compared to traditional amino carbohydrate columns, even with gradients Reduce run times with more mobile phase choices Compatible with gradient ELSD applications Quiet Gradient Baselines Prevail Carbohydrate ES Columns are excellent for isocratic separations with common detectors, but the column s real power shows with the ELSD and a solvent gradient. Enjoy the powerful selectivity, reduced run times, and efficient peaks from gradients while seeing the quiet, stable baselines that maximize sensitivity Min. Enhanced Detector Sensitivity Through Flat Gradient Baselines Min. Silica Based Amino Column 986 Prevail Carbohydrate ES Column (Part No. 0) Better Resolution and Peak Shape Prevail Carbohydrate ES Columns are versatile enough for mono- and oligosaccharides and sugar alcohols. They produce single peaks for reducing sugars at ambient temperatures, eliminating the need for column heating.. Dextrose. Maltose. Maltotriose. Maltotetraose. Maltopentaose 6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose 9. Maltononaose 0. Maltodecaose. Maltoundecaose. Maltododecaose. Maltotridecaose. Maltotetradecaose. Maltopentadecaose Column:.6 x 0mm Mobile Phase: A: Acetonitrile B: Water Gradient: Time: 0 %B: 0 Flow Rate:.0mL/min Detector: ELSD Use gradients with a Prevail Carbohydate ES column and the ELSD for shorter run times, better resolution, and greater sensitivity. By comparison, silica-based amino columns give noisy, shifting baselines. 6 7 Pepsi One Cola Min Sucralose. Unknown. Fructose. Glucose. Unknown 6. Sucrose 7. Maltose 8. Maltodextrins Column: Prevail Carbohydrate ES, µm,.6 x 0mm (Part No. 0) Mobile Phase: A: Acetonitrile B: Water Gradient: Time: 0 0 %B: Flow Rate:.0mL/min Detector: ELSD Column Temp: Ambient Prevail HPLC Columns Packing Format i.d. x Length Part No. Carbohydrate ES, Analytical.6 x 0mm 0 µm Analytical.6 x 0mm 0 Rocket 7 x mm 0 6 Robitussin Flu Syrup Min.. Sucralose. Fructose. Sorbitol. Glucose. Sucrose 6. Maltose 07 Column: Prevail Carbohydrate ES, µm,.6 x 0mm (Part No. 0) Mobile Phase: A: Acetonitrile B: Water Gradient: Time: 0 0 %B: 0 Flow Rate:.0mL/min Detector: ELSD Column Temp: Ambient Prevail All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Carbohydrate ES, µm.6 x 7.mm 96 All-Guard Cartridge Holder ea 800 *All-Guard holder required. Other particle sizes available. 7

25 Alltech Platinum Columns For Challenging Separations Unique selectivity Better peak shapes with polar analytes More separation choices with dual-selectivity Excellent stability and reproducibility.µm high throughput media for speed and resolution, especially when combined with Rocket and Expedite hardware The Platinum Column Advantage Controlled silica exposure is the difference that makes Platinum columns unique. Instead of thoroughly covering the silica with bonded phase to hide the silica, the exposure of the silica in Platinum columns is controlled to provide a dual mode separation with both polar and non-polar sites exposed to your samples. This extends polar selectivity well beyond what other reversed-phase columns offer and gives separations other columns cannot. Standard Platinum Columns vs Platinum EPS Columns Platinum columns come in two varieties offering different levels of silica exposure. Standard Platinum has a moderate silica exposure and is best used with neutral and moderately polar compounds. Platinum EPS (Extended Polar Selectivity) has a high level of silica exposure and is best used with compounds containing more than two polar functional groups. Alltech Platinum Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical.,, µm 00Å 00m /g 6% Monomeric Yes L EPS C8 Silica Spherical.,, µm 00Å 00m /g % Monomeric No L C8 Silica Spherical.,, µm 00Å 00m /g % Monomeric Yes L7 EPS C8 Silica Spherical, µm 00Å 00m /g.0% Monomeric No L7 Phenyl Silica Spherical, µm 00Å 00m /g Monomeric Yes L Cyano Silica Spherical, µm 00Å 00m /g Monomeric No L0 Amino (NH) Silica Spherical, µm 00Å 00m /g Monomeric No L8 Silica Silica Spherical, µm 00Å 00m /g L SAX Silica Spherical, µm* 00Å 00m /g Monomeric No Trying to solve difficult separation problems using typical reversed-phase columns often leads to the same result. Choose Platinum columns for completely different selectivity. See chart below. Hydrophobic/Hydrophillic Balance at ph of Various Reversed-Phase HPLC Packings.6.. Platinum EPS C8 Platinum EPS C8 k Amitripyline / k Toluene Standard Platinum C8 Typical RP C8 N O T E Platinum column s unique selectivity stands out from the competition. Standard Platinum C8 Typical RP C8 0 Typical Polar Embedded k Toluene Plotting k' values of different compounds (polar vs. nonpolar) demonstrate the unique selectivity of Platinum and Platinum EPS columns, compared to conventional reversed-phase columns. 8

26 Reverse Elution Order with Standard Platinum and Platinum EPS Columns Often, it s preferable when minor components elute before, rather than after, closely retained major components., Standard Platinum C Min. Platinum EPS C8 Vasodilators 887. Pindolol. Pentoxifylline. Isoxsuprine. Nifedipine Degradant. Nifedipine 888 Column: µm,.6 x 0mm Mobile Phase: 0.0M KHPO, ph.0:methanol (0:0) Flow Rate:.0mL/min Detector: UV at 0nm Platinum EPS Columns Have High Polar Compound Capacity This is important for early eluting polar compounds which are often unresolved on conventional reversed-phase columns. Capacity Factor Capacity Factors for Platinum EPS C8 Columns Biphenyl Amitriptyline Toluene Nitrobenzene Cinnamic Acid Consecutive Batches Platinum HPLC Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length C8,.µm Rocket 7.0 x mm 07 Rocket 7.0 x mm 09 C8, µm Solvent Reducer.0 x 0mm 79 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7.0 x mm 0 Rocket 7.0 x mm 0 C8, µm Solvent Reducer.0 x 0mm 79 Solvent Reducer.0 x 0mm 79 Analytical.6 x 0mm 0 0 Analytical.6 x 0mm EPS C8,.µm Rocket 7.0 x mm 077 Rocket 7.0 x mm 079 EPS C8, µm* Capillary 0.0x 0mm 00 Capillary 0.0x00mm 0 Capillary 0.0x0mm Capillary 0.00x 0mm Capillary 0.00x00mm Capillary 0.00x0mm Solvent Reducer.0 x 0mm 799 Analytical.6 x 00mm 8 6 Analytical.6 x 0mm 8 8 Rocket 7.0 x mm 07 Rocket 7.0 x mm 07 EPS C8, µm Solvent Reducer.0 x 0mm 806 Solvent Reducer.0 x 0mm 80 Analytical.6 x 0mm 6 Analytical.6 x 0mm 6 7 C8,.µm Rocket 7.0 x mm 09 C8, µm Rocket 7.0 x mm 0 C8, µm Analytical.6 x 0mm 70 7 Analytical.6 x 0mm 7 76 EPS C8, µm Analytical.6 x 0mm 6 Rocket 7.0 x mm 08 Rocket 7.0 x mm 08 EPS C8, µm Analytical.6 x 0mm 0 Analytical.6 x 0mm 6 *.µm and µm particles and other dimensions are available. Platinum HPLC Columns (continued) Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length Phenyl, µm Analytical.6 x 0mm 6 6 Phenyl, µm Analytical.6 x 0mm Analytical.6 x 0mm 6 6 Cyano, µm Rocket 7.0 x mm 09 Rocket 7.0 x mm 09 Cyano, µm Analytical.6 x 0mm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Rocket 7.0 x mm 0 Amino, µm Analytical.6 x 0mm 7 7 Analytical.6 x 0mm 7 7 Silica, µm Analytical.6 x 0mm 6 Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm 9 0 SAX, µm Analytical.6 x 0mm 9 9 SAX, µm Analytical.6 x 0mm 9 96 Analytical.6 x 0mm 9 9 Platinum Guard Cartridges Packing i.d. x Length Qty. Part No. C8 All-Guard, µm*.0 x 7.mm 99.6 x 7.mm 606 EPS C8, µm 0.0 x 0mm 69 Capillary Guard** 0.00 x 0mm 69 EPS C8 All-Guard, µm*.0 x 7.mm x 7.mm 607 C8 All-Guard, µm*.6 x 7.mm 6 EPS C8 All-Guard, µm*.6 x 7.mm 6 Phenyl All-Guard, µm*.6 x 7.mm 69 Cyano All-Guard, µm*.6 x 7.mm 60 Amino All-Guard, µm*.6 x 7.mm 6 Silica All-Guard, µm*.6 x 7.mm 6 SAX All-Guard, µm*.6 x 7.mm 787 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) Capillary Guard Cartridge Holder Guard Holder for 0.00mm and 0.0mm Guards ea GR-70E Guard Holder for 0.00mm and 0.00mm Guards ea GR-70A *All-Guard holder required. **.µm and µm particles and other dimensions are available. 9

27 GraceSmart Columns High Quality HPLC Phases at Exceptional Value High purity phases Efficient and reproducible separations General use selectivity Exceptional value GraceSmart HPLC phases use high-purity silica and are monomerically bonded with uniform coverage. This translates into symmetrical peaks for acids/bases, and predictable reversed-phase selectivity. Whether routine analysis or new method development, use GraceSmart columns to get premium performance at exceptional value. Reproducible Methods Start with Reproducible Columns Our tightly controlled silica synthesis and bonding keep capacity factor and selectivity variations to a minimum. The advanced packing methods deliver consistently high columnto-column efficiency. Every column is individually tested and shipped with a quality assurance chromatogram. best value new GraceSmart Phase Specifications Phase Encapping Surface Area Pore Size Particle Size C8 TMS, non-polar 0m /g 0Å, µm Expect Competitive Performance Six sigma techniques and lean manufacturing are key to our cost competitive position, without compromising quality or performance. GraceSmart columns show similiar or better efficiency and asymmetries for challenging base and acid components in comparison to industry leading columns. Capacity Factor Batch Toluene N-Butylbenzoic Acid Anisole Highly Reproducible Acetophenone P-Nitrophenol Amitriptyline Consistent batch-to-batch capacity for acidic, basic, and neutral compounds. Symmetry Plates / Columns,000 0,000 8,000 6,000,000, Nicotine Nicotine Efficiency Quinine Symmetry Quinine Diphenhydramine Diphenhydramine 0 Competitor C8 Column Competitor C8 Column Competitor C8 Column GraceSmart C8 Column GraceSmart columns compare favorably to leading competitor columns. Nortriptyline Nortriptyline, Dichlorophenoxy Acetic Acid, Dichlorophenoxy Acetic Acid, Dichlorophenoxy Propionic Acid, Dichlorophenoxy Propionic Acid, Dichlorophenoxy Butyric Acid, Dichlorophenoxy Butyric Acid GraceSmart RP8 Columns Packing Format i.d. x Length Part No. C8, µm Analytical.6 x 0mm 88 Analytical.6 x 0mm 880 Analytical. x 0mm 88 Analytical. x 0mm 88 C8, µm Analytical.6 x 0mm 7 Analytical.6 x 00mm 7 Analytical. x 0mm 7 Analytical. x 00mm 7 Analytical. x 0mm 76 Analytical.6 x 0mm 8 0

28 GraceSmart Columns mau Amoxicillin (U.S.P.) Min. GraceSmart C8 Column Luna C8 Column Inertsil ODS- Column Hypersil BDS C8 Column Kromasil 00- C8 Column Column: GraceSmart C8 Column, µm,.6 x 0mm (Part No. 880) Mobile Phase: Isocratic, 0.0N Monobasic Potassium Phosphate ph :Acetonitrile (96:) Flow Rate:.mL/min Detector: UV at 0nm Column Temp: 0 C Injection Vol: 0µL Min. 7 Standards Mix Ibuprofen (U.S.P.) Nicotine. Quinine. Diphenhydramine. Nortriptyline.,, Dichlorophenoxy Acetic Acid 6.,, Dichlorophenoxy Propionic Acid 7.,, Dichlorophenoxy Butyric Acid Column: GraceSmart RP8, μm,.6 x 0mm Mobile Phase: 0% MeCN/0% 0mM, KHPO, ph at C 070. Valerophenone. Ibuprofen Hypersil BDS C8 Kromasil 00- C8 USP GraceSmart Luna Inertsil Spec C8 C8 ODS- N/A Ret. Time (min) Theoretical plates Tailing Factor k'. k' Min. Column: GraceSmart C8 Column, µm,.6 x 0mm (Part No. 88) Mobile Phase: HO (adjusted with Phosphoric Acid to a ph of.): Acetonitrile (6:) Flow Rate: ml/min Detector: UV at nm Injection Vol: µl Carbamates Min Fenuron. Monuron. Fluometuron. Diuron. Methiocarb 6. Linuron 7. Neburon Column: GraceSmart C8 Column, µm,.6 x 0mm (Part No. 88) Mobile Phase: A: 0.0M KHPO, ph. B: Acetonitrile Gradient: Time: 0 0 %B: 0 60 Flow Rate:.0mL/min Detector: UV at 0nm Injection Vol: 0µL more applications To view our complete searchable chromatogram database visit /chromdb/ Min. Column: 07. Ascorbic Acid. Nicotinic Acid. Pyridoxamine. Niacinamide. Folic Acid 6. Vitamin B 7. Riboflavin GraceSmart C8 Column, μm,.6 x 0mm (Part No. 88) Mobile Phase: A: 0.M NHHPO Buffer, ph. B: Acetonitrile Gradient: Time: 0 0 %B: 0 0 Flow Rate: Detector: Injection Vol: Water Soluble Vitamins.0mL/min UV at nm μl

29 GraceAlpha Columns (Patent Pending) A New Silica Generation High-porosity surface increases mass transfer and results in increased column efficiencies and loading capacity Dense core and highly spherical shape yields mechanically robust particle Proprietary bonding technology gives unique selectivity and increased resolution Ideally suited for scale up to prep applications Extreme Porosity High-porosity exterior increases mass transfer leading to higher efficiencies and superior loading capacity. The high-porosity structure adds an elastic quality to the silica surface allowing the particles to pack tighter yielding higher efficiencies. It also stabilizes the packed bed preventing voids. Dense Inner Core new Dense core provides exceptional strength. Typically the high-porosity structure would result in a weak particle prone to breaking, but the GraceAlpha particle combines the high porosity outer region with a dense inner core that results in a particle that resists breaking and forming fines that contribute to high backpressures. GraceAlpha Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical, 0,, 0µm 0Å m /g % Monomeric Yes L C8 Silica Spherical, 0,, 0µm 0Å m /g 0% Monomeric L7 Silica Silica Spherical, 0,, 0µm 0Å m /g Monomeric L High Level of Resolution GraceAlpha offers resolution advantages due to its higher mass transfer and optimized C8 and C8 bonding chemistry. Adsorbence Adsorbence Crude 0-AA Synthetic Peptide from Biopeptide Co., Inc. Better resolution of closely eluting impurities GraceAlpha Column µm Competitor µm Min Min. GraceAlpha Columns Packing i.d. x Length Part No. C8, µm.6 x 0mm 07.6 x 0mm 070 C8, 0µm.6 x 0mm 07.6 x 0mm 070 C8, µm.6 x 0mm 08.6 x 0mm 09 C8, µm.6 x 0mm x 0mm 070 C8, 0µm.6 x 0mm 07.6 x 0mm 070 C8, µm.6 x 0mm 0.6 x 0mm 07 Silica, µm.6 x 0mm x 0mm 070 Silica, 0µm.6 x 0mm 07.6 x 0mm 070 Silica, µm.6 x 0mm 0.6 x 0mm 07 Columns: All C8.6 x 0mm Mobile Phase: A: 0.% TFA in Water B: 0.% TFA in Acetonitrile Gradient: Time: 0 0 %B: 0 0 Flow Rate:.0mL/min The target peptide is better resolved from impurities, with % higher recovery, on the GraceAlpha column. more applications To view our complete searchable chromatogram database visit /chromdb/

30 Alltech Brava Columns If You Like Hypersil BDS, Try This Lower-Cost Alternative BDS phases for acids, bases, and neutrals ODS phases for non-polars and lipophilics Medium-sized pores are ideal for resolution of small- to medium-sized compounds, while its less-retentive nature means less mobile phase consumption. Brava HPLC Columns Packing i.d. x Length Part No. Waters Fittings Part No. C8 BDS, µm.6 x 00mm x 0mm C8 BDS, µm.6 x 0mm x 0mm C8 ODS, µm.6 x 00mm x 0mm C8 ODS, µm.6 x 0mm x 0mm C8 BDS, µm.6 x 00mm x 0mm C8 BDS, µm.6 x 0mm x 0mm C8, µm.6 x 00mm x 0mm C8, µm.6 x 0mm x 0mm Phenyl, µm.6 x 0mm x 0mm Cyano BDS, µm.6 x 0mm x 0mm Cyano, µm.6 x 0mm x 0mm Amino, µm.6 x 0mm x 0mm Silica, µm.6 x 0mm x 0mm best value Brava Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 BDS Silica Spherical, µm Å 8m /g 8.% Monomeric Yes L C8 ODS Silica Spherical, µm 0Å 9m /g 8.% Monomeric Yes L C8 Silica Spherical, µm 0Å 9m /g 6% Monomeric Yes L7 C8 BDS Silica Spherical, µm Å 8m /g.% Monomeric Yes L7 Phenyl Silica Spherical µm 0Å 9m /g Monomeric No L Cyano Silica Spherical µm 0Å 9m /g Monomeric No L0 Cyano BDS Silica Spherical µm Å 8m /g Monomeric No L0 Amino (NH) Silica Spherical µm 0Å 9m /g Monomeric No L8 Silica Silica Spherical µm 0Å 9m /g No L 0 Metal Chelator Min. Column: Brava C8 BDS, µm,.6 x 0mm (Part No. 09) Mobile Phase: Acetonitrile:0mM KHPO ph (0:0) Flow Rate:.0mL/min Detector: UV at nm 07. Pyridine., Dipyridyl Brava All-Guard Cartridges* Packing i.d. x Length Qty. Part No. C8 BDS, µm.6 x 7.mm 967 C8 ODS, µm.6 x 7.mm 9677 C8 BDS, µm.6 x 7.mm 967 C8, µm.6 x 7.mm 9678 Phenyl, µm.6 x 7.mm 9680 Cyano BDS, µm.6 x 7.mm 967 Cyano, µm.6 x 7.mm 9679 Amino, µm.6 x 7.mm 967 Silica, µm.6 x 7.mm 9676 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. more applications To view our complete searchable chromatogram database visit /chromdb/

31 Alltech Alltima Introduction High-Quality General-Purpose HPLC Columns Extensive collection of published applications Well suited for microbore applications, stable in popular microbore mobile phases Acid and base deactivated symmetrical peaks Polymerically bonded and double-endcapped for long column lifetimes Analyze acids, bases, and neutrals in one run Full range of column dimensions, from microbore to preparative Alltima Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical,, 0µm 00Å 0m /g 6% Polymeric Yes L C8 LL Silica Spherical µm 00Å 0m /g 9% Polymeric Yes L C8 Silica Spherical,, 0µm 00Å 0m /g 9% Polymeric Yes L7 Phenyl Silica Spherical, µm 00Å 0m /g 7.% Polymeric Yes L Cyano Silica Spherical, µm 00Å 0m /g Polymeric Yes L0 Amino (NH) Silica Spherical, µm 00Å 0m /g Polymeric No L8 Silica Silica Spherical,, 0µm 00Å 0m /g L Basic Drugs 89. Procainamide. Acetaminophen. Theophylline. Caffeine best seller Antibacterials 876. Oxytetracycline Dihydrate. Tetracycline HCL. Meclocycline Sulfosalicylate Salt 0 0 Min. Column: Alltima C8, µm,.6 x 0mm Mobile Phase: Methanol:0mM KHPO, ph.0 (0:80) Flow Rate:.0mL/min Detector: UV at nm Min. Column: Alltima C8, µm,.6 x 00mm Mobile Phase: A: 0mM KHPO, ph.0 B: CHCN Gradient: Time: 0 0 %B: 0 Flow Rate:.0mL/min Detector: UV at nm Natural Products 8887 Antihistamines 7. Naringin. Neohesperidin. Myricetin. Quercetin. Naringenin 6. Kaempferol 6. Tripelennamine. Triprolidine. Cyclizine. Chlorcyclizine. Meclizine 0 0 Min. Column: Alltima C8, µm,.6 x 00mm Mobile Phase: A: 0mM KHPO, ph. B: CHCN Gradient: Time: 0 %B: 0 0 Flow Rate:.0mL/min Detector: UV at 80nm Min. Column: Alltima C8, µm,.6 x 0mm Mobile Phase: A: 0mM KHPO, ph. B: CHCN Gradient: Time: 0 0 %B: 70 Flow Rate:.mL/min Detector: UV at 0nm

32 Alltech Alltima Reversed-Phase Columns 0 7, 6 Taxol and Related Taxanes in Yew (taxus brevifolia) deacetylbaccatin III. Baccatin III. 7-xylosyl-0-deacetyltaxol B. Taxinine M. 7-xylosyl-0-deacetyltaxol 6. 7-xylosyl-0-deacetyltaxol C 7. 0-deacetyltaxol 8. 7-xylosyltaxol 9. Cephalomannine (Taxol B) 0. 7-epi-0-deacetyltaxol. Taxol. Taxol C. 7-epitaxol 8 Min. Column: Alltima Phenyl Rocket, µm, 7 x mm Mobile Phase: 0.0% TFA in HO:MeOH:CHCN (0:9:) Flow Rate:.mL/min Detector: UV at 0nm Anticonvulsants Min. Column: Alltima Phenyl, µm,.6 x 0mm Mobile Phase: A: HO B: CHCN Gradient: Time: %B: Flow Rate:.mL/min Detector: UV at nm Ethotoin. Phenobarbital. Methsuximide. Mephenytoin. Carbamazepine 6. Diazepam Alltima All-Guard Cartridges* Packing i.d. x Length Qty. Part No. C8, µm. x 7.mm x 7.mm x 7.mm C8-LL, µm. x 7.mm 96.0 x 7.mm 96.6 x 7.mm 968 C8, µm. x 7.mm 96.0 x 7.mm x 7.mm 9608 Phenyl, µm. x 7.mm 96.0 x 7.mm 96.6 x 7.mm 9608 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. Alltima HPLC Reversed-Phase Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length C8, µm Microbore.0 x 00mm 8 Microbore.0 x 0mm 86 Expedite MS. x 0mm 86 Expedite MS. x 0mm 80 Microbore. x 0mm 89 Microbore. x 00mm 8 Microbore. x 0mm 8 Solvent Reducer.0 x 0mm 8 Expedite MS.6 x 0mm 87 Expedite MS.6 x 0mm 8 Analytical.6 x 0mm 8 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 060 Rocket 7 x mm 060 C8, µm Microbore.0 x 0mm 888 Microbore.0 x 0mm 888 Microbore. x 0mm 8870 Microbore. x 0mm 887 Solvent Reducer.0 x 0mm 80 Solvent Reducer.0 x 0mm 8 Analytical.6 x 0mm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm C8-LL, µm Microbore.0 x 0mm 8890 Microbore.0 x 0mm 889 Microbore. x 0mm 8888 Microbore. x 0mm 8889 Solvent Reducer.0 x 0mm 8 Solvent Reducer.0 x 0mm 8 Analytical.6 x 0mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm 8 Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 0607 Rocket 7 x mm 0609 C8, µm Microbore.0 x 0mm 889 Microbore.0 x 0mm 889 Microbore. x 0mm 887 Microbore. x 0mm 887 Solvent Reducer.0 x 0mm 86 Analytical.6 x 0mm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm Phenyl, µm Microbore.0 x 00mm 89 Microbore.0 x 0mm 8 Expedite MS. x 0mm 809 Expedite MS. x 0mm 86 Microbore. x 0mm 8 Microbore. x 00mm 866 Microbore. x 0mm 8 Expedite MS.6 x 0mm 88 Expedite MS.6 x 0mm 8 Analytical.6 x 0mm 878 Analytical.6 x 00mm Analytical.6 x 0mm Phenyl, µm Solvent Reducer.0 x 0mm 87 Analytical.6 x 0mm Analytical.6 x 0mm

33 Alltech Alltima C8 Validation Kits Alltima columns have the batch-to-batch and column-tocolumn reproducibility required for method development. Each kit includes three columns made from three different batches of Alltima C8. Alltima is Stable and Reproducible Initial Separation 7 Alltima C8 Validation Kits Description Includes: i.d. x Length Part No. C8, µm columns from different batches.6 x 00mm 0 C8, µm columns from different batches.6 x 0mm C8, µm columns from different batches.6 x 0mm Alltech Alltima Normal-Phase Columns Alltima HPLC Normal-Phase Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length Cyano, µm Analytical.6 x 0mm 879 Analytical.6 x 00mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm 880 Analytical.6 x 00mm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Analytical.6 x 0mm Silica, µm Microbore.0 x 00mm 8 Microbore.0 x 0mm 8 Expedite MS. x 0mm 89 Expedite MS. x 0mm 80 Microbore. x 0mm 876 Microbore. x 00mm 8 Microbore. x 0mm 88 Expedite MS.6 x 0mm 889 Expedite MS.6 x 0mm 886 Analytical.6 x 0mm 8 Analytical.6 x 00mm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm Silica, 0µm Analytical.6 x 0mm Alltima All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Cyano, µm.6 x 7.mm 9608 Amino, µm.6 x 7.mm 9608 Silica, µm. x 7.mm x 7.mm 9608 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available Min. After 00 Hours of Washing at 60 C Min. Column: Alltima C8, µm,.6 x 0mm Mobile Phase: A: Water:Acetonitrile:TFA (9::0.) B: Water:Acetonitrile:TFA (:7:0.) Gradient: Time: 0 60 % B: 0 Flow Rate: ml/min Detector: UV at 0nm Column capacity and selectivity remained constant after washing an Alltima C8 column for 00 hours with 0.% TFA:Acetonitrile (0:0) at 60 C. 7. Gly-Tyr. Val-Tyr-Val. Met-Enkephalin. Leu-Enkephalin. Angiotensin II tech tip Want high throughput on a conventional (non-ultra high-pressure) system? Choose the Rocket column format no modifications necessary to your existing HPLC system. Choose Expedite columns for low volume and microbore systems. See page for more explanation of the Rocket and Expedite formats. 6

34 Alltech Apollo Columns Economical, High Performance Columns Easy scale-up from analytical to prep Extended ph stability. to 0. Batch-tested Apollo HPLC columns use high purity, basedeactivated silica for powerful separations at an economical price. They are ideal for educational labs or for method development. Apollo Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical µm 00Å 0m /g % Monomeric Yes L C8 Silica Spherical µm 00Å 0m /g 9% Monomeric Yes L7 Phenyl Silica Spherical µm 00Å 0m /g 8% Monomeric Yes L Silica Silica Spherical µm 00Å 0m /g L Apollo HPLC Columns Packing Format i.d. x Length Part No. C8, µm Analytical.6 x 0mm 60 Analytical.6 x 0mm 6 C8, µm Analytical.6 x 0mm 606 Analytical.6 x 0mm 6 Phenyl, µm Analytical.6 x 0mm 68 Analytical.6 x 0mm 6 Silica, µm Analytical.6 x 0mm 607 Analytical.6 x 0mm 6 best value Apollo All-Guard Cartridges* Packing i.d. x Length Qty. Part No. C8, µm.6 x 7.mm 96 C8, µm.6 x 7.mm 966 Phenyl, µm.6 x 7.mm 960 Silica, µm.6 x 7.mm 969 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. Alltech Econosphere Columns Low-cost columns for common chromatography applications Acid-washed for low metal content Easy scale-up from analytical to prep Econosphere Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical,, 0µm 80Å 00m /g 0% Monomeric Yes L C8 Silica Spherical,, 0µm 80Å 00m /g % Monomeric Yes L7 Cyano Silica Spherical µm 80Å 00m /g Monomeric Yes L0 Amino (NH) Silica Spherical µm 80Å 00m /g Polymeric Yes L8 Silica Silica Spherical,, 0µm 80Å 00m /g No L Econosphere HPLC Columns Standard Part No. Waters Part No. Packing Format i.d. x Length C8, µm Analytical.6 x 0mm Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 067 Rocket 7 x mm 067 C8, µm Analytical.6 x 0mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 00mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm Analytical.6 x 0mm Econosphere HPLC Columns (continued) Standard Part No. Waters Part No. Packing Format i.d. x Length Amino, µm Analytical.6 x 0mm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm Analytical.6 x 00mm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm Econosphere All-Guard Cartridges* Packing i.d. x Length Qty. Part No. C8.6 x 7.mm 96 C8.6 x 7.mm 96 Cyano.6 x 7.mm 96 Amino.6 x 7.mm 96 Silica.6 x 7.mm 96 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *Guard holder required. 7

35 Alltech Adsorbosphere Columns Reliable performance from rugged, reproducible phases High temperature bonding for exceptional surface coverage, stability, and reproducibility. Adsorbosphere columns are the workhorses of any chromatography lab. Choose from four varieties: Adsorbosphere has a moderate carbon load for common applications. Adsorbosphere HS has a larger surface area and higher carbon load for stronger retention of hydrophobic compounds. Adsorbosphere UHS has the largest surface area and highest carbon load for the highest resolution of hydrophobic compounds. Adsorbosphere XL is a lower-cost, larger pore alternative. C8-B separates strongly basic analytes more efficiently then standard C8. Adsorbosphere Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code Adsorbosphere C8 Silica Spherical,, 0µm 80Å 00m /g % Monomeric Yes L Adsorbosphere C8 HS Silica Spherical, µm 60Å 0m /g 0% Monomeric Yes L Adsorbosphere C8 UHS Silica Spherical, 0µm 60Å 00m /g 0% Monomeric Yes L Adsorbosphere C8 Silica Spherical µm 80Å 00m /g 8% Monomeric Yes L7 Adsorbosphere Phenyl Silica Spherical µm 80Å 00m /g % Monomeric Yes L Adsorbosphere Cyano Silica Spherical µm 80Å 00m /g Monomeric Yes L0 Adsorbosphere Cyano-AQ Silica Spherical µm 0Å 70m /g Polymeric No L0 Adsorbosphere Amino (NH) Silica Spherical, µm 80Å 00m /g Polymeric No L8 Adsorbosphere Silica Silica Spherical µm 80Å 00m /g L Adsorbosphere SAX Silica Spherical µm 80Å 00m /g Monomeric Adsorbosphere SCX Silica Spherical µm 80Å 00m /g Monomeric Yes Adsorbosphere XL C8 Silica Spherical, µm 90Å 00m /g % Monomeric Yes L Adsorbosphere XL C8-B Silica Spherical µm 90Å 00m /g % Monomeric Yes L Adsorbosphere XL C8 Silica Spherical, µm 90Å 00m /g 6% Monomeric Yes L7 Adsorbosphere XL C (TMS) Silica Spherical µm 90Å 00m /g Monomeric Yes L Adsorbosphere XL Silica Silica Spherical µm 90Å 00m /g L Adsorbosphere XL SAX Silica Spherical, 0µm 90Å 00m /g Monomeric Yes Adsorbosphere XL SCX Silica Spherical, 0µm 90Å 00m /g Monomeric Yes Min. Column: Mobile Phase: Flow Rate: Detector: Column Temp: Vitamins D & D 0. Vitamin D. Vitamin D Adsorbosphere UHS C8, µm,.6 x 0mm (Part No. 888) 80% Acetonitrile 0% Methanol.mL/min UV at nm Ambient Adsorbosphere HPLC Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length C8, µm Analytical.6 x 00mm Rocket 7 x mm 06 Rocket 7 x mm 06 C8, µm Analytical.6 x 0mm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 0mm Phenyl, µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm Cyano-AQ, µm Analytical.6 x 0mm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm SAX, µm Analytical.6 x 0mm SCX, µm Analytical.6 x 0mm Adsorbosphere HS HPLC Columns Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length C8 HS, µm Analytical.6 x 00mm Analytical.6 x 0mm C8 HS, µm Analytical.6 x 0mm Analytical.6 x 0mm

36 Alltech Adsorbosphere Columns (continued) Adsorbosphere UHS HPLC Columns Adsorbosphere XL HPLC Columns Standard Part No. Standard Part No. Waters Fittings Part No. Packing Format i.d. x Length C8 UHS, µm Analytical.6 x 0mm C8 UHS, 0µm Analytical.6 x 0mm Waters Fittings Part No. Packing Format i.d. x Length XL C8, µm Analytical.6 x 00mm Analytical.6 x 0mm Rocket 7 x mm 06 Rocket 7 x mm 06 XL C8, µm Analytical.6 x 0mm Analytical.6 x 0mm XL C8-B, µm Analytical.6 x 0mm 08 0 Analytical.6 x 0mm XL C8, µm Analytical.6 x 0mm XL C8, µm Analytical.6 x 0mm Analytical.6 x 0mm XL C, µm Analytical.6 x 0mm Analytical.6 x 0mm XL Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm XL SAX, µm Analytical.6 x 0mm Analytical.6 x 0mm XL SAX, 0µm Analytical.6 x 0mm 0 00 XL SCX, µm Analytical.6 x 0mm 0 0 Analytical.6 x 0mm 0 06 XL SCX, 0µm Analytical.6 x 0mm Alltech Allsphere Columns Try Alltech s lower cost Allsphere columns as a replacement for Spherisorb columns. If you are not satisfied with Allsphere as a direct replacement for your Spherisorb applications, we will refund your money. Allsphere HPLC Columns Standard Part No. Waters Fittings Part No. Packing i.d. x Length ODS-, µm.6 x 0mm x 0mm ODS-, µm.6 x 00mm x 0mm ODS-, µm.6 x 0mm x 0mm C8, µm.6 x 00mm x 0mm C8, µm.6 x 0mm x 0mm C6, µm.6 x 0mm x 0mm Phenyl, µm.6 x 0mm x 0mm Cyano, µm.6 x 0mm x 0mm Amino, µm.6 x 0mm x 0mm Silica, µm.6 x 00mm x 0mm Silica, µm.6 x 0mm x 0mm SAX, µm.6 x 0mm SCX, µm.6 x 0mm Adsorbosphere All-Guard Cartridges* Packing i.d.x Length Qty. Part No. Amino, µm.6 x 7.mm 960 C8, µm.6 x 7.mm 960 C8 HS, µm.6 x 7.mm C8 UHS, µm.6 x 7.mm 88 C8, µm.6 x 7.mm 960 Cyano, µm.6 x 7.mm 9607 Cyano-AQ, µm.6 x 7.mm 9608 Phenyl, µm.6 x 7.mm 960 Silica, µm.6 x 7.mm 9606 SAX, µm.6 x 7.mm 9609 SCX, µm.6 x 7.mm 9607 XL C8, µm.6 x 7.mm XL C8-B, µm.6 x 7.mm XL C8, µm.6 x 7.mm XL C, µm.6 x 7.mm 9609 XL Silica, µm.6 x 7.mm XL SAX, µm.6 x 7.mm XL SCX, µm.6 x 7.mm All-Guard Cartridge Holder (Includes Direct-Connect Column Coupler) ea 800 *All-Guard holder required. Other particle sizes available. Allsphere All-Guard Cartridges* Packing i.d. x Length Qty. Part No. ODS-, µm.6 x 7.mm 960 ODS-, µm.6 x 7.mm 960 C8, µm.6 x 7.mm 960 C6, µm.6 x 7.mm 960 Phenyl, µm.6 x 7.mm 9606 Cyano, µm.6 x 7.mm 9608 Amino, µm.6 x 7.mm 9609 Silica, µm.6 x 7.mm 960 SAX, µm.6 x 7.mm 960 SCX, µm.6 x 7.mm 96 All-Guard Cartridge Holder (Includes Direct-Connect Column Coupler) ea 800 *All-Guard holder required. Other particle sizes available. more info Looking for Adsorbosil columns? Please visit. 9

37 Grom Sil Columns High-Quality, Wide Variety, HPLC Columns for All Types of Applications Highly consistent manufacturing for excellent reproducibility 0 different phases to choose from, general purpose to specialty phases Wide range of particle sizes from sub μm to 0µm Available in nano to preparative column formats High purity spherical silica with high and universal selectivity for broad range of applications. Different phases and bonding technologies give you the power and flexibility ST = standard bonding with endcapping high-quality universal column NE = not endcapped high polar selectivity HE = hydrophilic endcapping increased polar selectivity; ph-stable at high ph, high carbon load and very low silanol activity CP = possiblility to work at high ph FE = Fully endcapped maximum silica coverage PA = Cross-linked polyamino better stability ph = ph stable use mobile phases up to ph Grom Sil Phase Specifications Base Particle Material Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? Phase ODS- CP (encapsulated) Silica Spherical,, 7, 0µm 0, 00Å 0, 70m /g, 6% Polymeric No L ODS- HE (hydrophilic endcapping) Silica Spherical,,, 7, 0µm 0, 00Å 00, 00m /g 6, % Monomeric Yes L ODS- ST (standard) Silica Spherical,,, 7, 0µm 60, 0, 00Å 80, 00, 00m /g, 7, % Monomeric Yes L ODS-7 ph (ph-stable) Silica Irregular µm 80Å 0m /g % Polymeric No L Octyl- CP (encapsulated) Silica Spherical,, 7, 0µm 0, 00Å 0, 70m /g 0,.% Polymeric No L7 Phenyl- FE (fully endcapped) Silica Spherical,, 0µm 0, 00Å 00, 0m /g 9, % Monomeric Yes L Phenyl- CP (encapsulated) Silica Irregular µm 0, 00Å 0, 70m /g 7, % Polymeric No L Cyano- CP (encapsulated) Silica Spherical µm 0Å 0m /g % Polymeric No Amino- PA (Crs linked Poly-NH) Silica Spherical µm 0Å 00m /g Polymeric No L8 Diol Silica Spherical, 0µm 60, 0, 00Å 80, 00, 00m /g Monomeric No L0 USP L-code 0 Color Additives Dyes 6 7. Tartrazine. Amaranth. Indigo Carmine. New Coccine. Sunset Yellow 6. Fast Green 7. Brilliant Blue 8. Erythosine 9. Acid Red 0. Phloxine. Rosé Bengale Column: Grom Sil 0 ODS- ST, µm, x 0mm Flow Rate: 0.8mL/min Mobile Phase: A: 0mM NH-phosphate, ph 6.0:MeOH (90:0) B: 0mM NH-phosphate, ph 6.0:MeOH (0:0) Gradient: 0 00% B (0 0min, linear), 00% B (0 60min) Column Temp: 0 C Detector: UV at nm Min Min. Nucleosides and Bases. Cytosine. Cytidine. Uracil. Deoxycytidine. Guanine 6. Adenine 7. Hypoxanthine 8. Uridine 9. Xanthine 0. Thymine. Inosine. Guanosine. Deoxyinosine. Deoxyguanosine. Xanthosine 6. Thymidine 7. Adenosine 8. Deoxyadenosine Column: Grom Sil 0 ODS- ST, μm, x 0mm Flow Rate: 0.mL/min Mobile Phase: A: 0mM NH-acetate, ph. B: 0mM NH-acetate, ph.:meoh (90:0) Gradient: Time: 0 0 %B: Detector: UV at 60nm, 0.08 AUFS 60

38 Grom Sil Columns 0 Water-soluble Vitamins 6 Column: Grom Sil 0 ODS- ST, µm, x 0mm (Part No. 6) Flow Rate: 0.8mL/min Mobile Phase: A: 0mM K-phosphate, ph 6.6 B: 0mM K-phosphate, ph 6.6:ACN (80:0) Gradient: Time: 0 0 %B: Column Temp: 0 C Detector: UV at nm Min. 6 7 Fat-soluble Vitamins 8 9. Menadione (Vit. K). Retinol (Vit. A). Retinol Acetate. Menaquinone (Vit. K). d-tocopherol 6. Ergocalciferol (Vit. D) 7. Cholecalciferol (Vit. D), 8. a-tocopherol (Vit. E), 9. a-tocopherol Acetate, 0. Phylloquinone (Vit. K) Column: Grom Sil 0 ODS- ST, µm, x 0mm (Part No. 6) Flow Rate: 0.8mL/min Mobile Phase: Acetonitrile Column Temp: 0 C Detector: UV at 80nm Min L-Ascorbic Acid (Vit. C). Nicotinic Acid. Pyridoxine HCl (Vit. B6). Nicotinamide. Thiamine Hydrochloride (Vit. B) 6. Folic Acid 7. Cyanocobalamin (Vit. B) 8. Riboflavin Grom Sil Columns Packing Format i.d. x Length Part No. ODS- CP, µm Microbore x 60mm 7 Microbore x mm 67 Microbore x 0mm 706 Solvent Reducer x 60mm 97 Solvent Reducer x 00mm 798 Solvent Reducer x 0mm 60 Analytical x 60mm 98 Analytical.6 x 0mm 700 ODS- CP, µm Solvent Reducer x mm 60 Analytical x mm 79 Analytical.6 x 0mm 609 ODS- HE, 0µm Analytical.6 x 0mm 6 ODS- HE, µm Capillary 0. x 0mm 88 Microbore x 0mm 6060 Microbore x 0mm 69 Microbore x 00mm 97 Microbore x mm 6880 Microbore x 0mm 808 Analytical x 0mm 889 Analytical x 60mm 786 Analytical x mm 68 Analytical.6 x mm 77 ODS- HE, µm Capillary 0. x 0mm 7667 Capillary 0. x 0mm 909 Capillary 0. x 0mm 8 Microbore x mm 6 Microbore x 0mm 60 Solvent Reducer x 0mm 777 Analytical x mm 808 Analytical x 0mm 66 Analytical.6 x 0mm 6777 ODS- ST, µm Microbore x 60 mm 70 Microbore x 0mm 8 ODS- ST, µm Microbore x 0mm 6 Analytical x mm 69 Analytical x 0mm 6 Analytical x 0mm 66 Analytical.6 x 0mm 67 ODS-7 ph, µm Capillary 0. x 0mm mm Guard Column Microbore x 0mm 8 Microbore x 0mm 66 Solvent Reducer x 0mm 696 Analytical.6 x mm 6 Analytical.6 x 0mm 80 Octyl- HE, µm Analytical.6 x mm 6 Octyl- CP, µm Microbore x 0mm 7 Analytical x 0mm 76 Phenyl- FE, µm Microbore x 00mm 69 Analytical x 0mm 696 Phenyl- CP, µm Analytical x 0mm 69 Cyano- CP, µm Microbore x mm 70 Analytical.6 x 60mm 6 Amino- PA, µm Microbore x 0mm 69 Analytical x 0mm 66 Analytical.6 x 0mm 06 Diol, µm Microbore x 00mm 776 more applications To view our complete searchable chromatogram database visit /chromdb/ more info For large molecule columns, refer to page

39 Grom Sapphire Column High Purity for Optimal Performance of Challenging Separations Excellent peak shape Highly symmetrical peaks from high purity silica base High speed and high capacity Range of pore sizes and surface area for improved speed and resolution Optimized for consistent flow critical in low volume applications Variety of column formats.,, and µm particles Sapphire Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical,, 0µm 6, 0Å 00, 70m /g, 6% Monomeric Yes L C8 Silica Spherical,, 0µm 6, 0Å 00, 70m /g, 0% Monomeric Yes L7 C* Silica Spherical,, 0µm 6, 0Å 00, 70m /g 0., 7% Monomeric Yes L6 Silica* Silica Spherical,, 0µm 6, 0Å 00, 70m /g No L *Columns available upon request. Separation of Antiepileptics Ethosuximide. Primidone. Lamotrigine. Phenobarbital. Carbamazepine-0,-epoxide 6. IS, Hexobarbital 7. Phenytoin 8. Carbamazepine Stability of Grom Sapphire Packings (Daldrup) 000 mv 00. Uracil. -Hydroxybenzoic Acid. Diphenylhydramine. -(p-methylphenyl)--phenylhydantoin. Diazepam 6. Toluene Injection 00 Injection 0 Min. Column: Grom Sapphire 0 C8, µm, x mm (Part No. 799) Flow Rate:.mL/min Mobile Phase: 0mM Na-phosphate, ph 6.:MeOH:ACN (6::) Detector: UV at 0nm Sapphire HPLC Columns Packing i.d. x Length Part No. C8, µm 0.00 x 0mm x 0mm x 00mm 80 C8, µm x mm 799 x mm 70 C8, µm x 0mm 89 0 Injection Min. Column: Grom Sapphire 0 C8, µm, x mm (Part No. 70) Flow Rate: ml/min Mobile Phase: 0mM Na-phosphate, ph.:acn (8:) Column Temp: RT Detector: UV at 0nm technical assistance Contact Tech Support: Phone: (North America) contact.alltech@grace.com Online: related products Looking for HPLC column prefilters? See page. related products Need high-pressure polymeric fittings? See pages for our full selection of high-pressure fittings

40 Grom Amino Acid Column Kits Amino Acid Analysis by Precolumn Derivatization Using OPA/IBLC High resolution (both optical antipodes of all protein amino acids in a single chromatogram) Outstandingly high sensitivity (fmol) Extraordinarily long lifetimes of columns. L-Asp. D-Asp. L-Glu. D-GLu. L-Ser 6. D-Ser 7. L-Thr OPA/IBLC Derivatized D- and L-Amino Acids Column: Grom Sil OPA-, x 0mm Flow Rate: 0.8mL/min Mobile Phase: A: mm Na-acetate, ph 6.0 B: 600mL MeOH + 0mL ACN Gradient: 0 % B (0 7min) Column Temp: C Detector: Fluorescence 0nm(exc.), nm(em.) Min. 8. Gly 9. D-Thr 0. L-His. D-His. L-Ala. L-Arg. D-Arg. D-Ala 6. L-Tyr 7. D-Tyr 8. L-Val 9. L-Met 0. L-Nor-Val (Int. Standard). D-Met. D-Val. L-Phe. L-Ile. D-Phe 6. L-Leu 7. D-Ile 8. D-Leu 9. L-Lys 0. D-Lys Amino Acid Analysis by Precolumn Derivatization With OPA/-MPA Highest sensitivity ( 0fMol) Short analysis time Fully automated derivatization Best resolution Amino Acid Analysis after Precolumn Derivatization with OPA/Mercaptopropionic Acid Extended Standard Asp. Glu. Asn. Ser Min.. Gin 6. Gly 7. Thr 8. His 9. Cit 0. m-his. Ala. Tau. Arg. AAB. Tyr 6. Val 7. Met 8. Nor-Val 9. Trp 0. Ile. Phe. Leu. Orn. Lys Column: Grom Sil OPA-, x 0mm Mobile Phase: A: mm Na-phosphate, ph 7.:THF (99:) B: mm Na-phosphate, ph 7.:MeOH/ACN (0::) Gradient: Time: %B: Flow Rate: 0.8mL/min Column Temp: C Detector: Fluorescence 0nm(exc.), 0nm(em.) OPA/IBLC Kit Description Complete kit includes.0mm i.d. x 0mm length column, OPA/IBLC derivitization reagents, and detailed working instructions. Part No OPA/-MPA Kit Description Complete kit includes.6mm i.d. x 0mm length column, OPA/-MPA derivitization reagents, and detailed working instructions. Part No Amino Acid Analysis by Precolumn Derivatization Using FLEC/ADAM High resolution and high sensitivity Rapid, automated or manual derivatization Primary and secondary amines Stable derivatives FLEC/ADAM Kit Description Complete kit includes.0mm i.d. x 0mm length column, FLEC/ADAM derivitization reagents, and detailed working instructions. Part No Separation of a D-, L-Amino Acid Standard by HPLC Employing the FLEC-ADAM Approach. L-Arg. D-Arg. L-Ser. D-Ser. D-Asp 6. L-Asp 7. D-Thr 8. L-Thr 9. D-Glu 0. L-Glu. D,L-Pro. D-Ala. L-Ala. FLEC-OH. D-Tyr 6. D-Met 7. L-Tyr 8. L-Met 9. D-Val 0. D-Phe Min.. L-Val. L-Phe. D-allo-lie. D-lle. D-Leu 6. L-allo-lie; L-lie 7. L-Leu Column: Flow Rate: Mobile Phase: Gradient: Time: %B: %C: D-His 9. D-Om 0. L-His. D-Lys Grom Sil FLEC-, x 0mm 0.7mL/min A: 0mM Na-acetate, ph.0 B: ACN C: THF Column Temp: 0 C Detector: Fluorescence 6nm(exc.), nm(em.). L-Om. L-Lys 6

41 Jones Genesis Columns Genesis columns use a new generation adsorbent based on high-purity metal-free 0Å spherical silica. Particle size offering includes,, 7, and µm. Genesis columns exhibit excellent peak symmetry and exceptional ph stability from to 0. Genesis Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 Silica Spherical,, 7, µm 0Å 00m /g 8% Monomeric Yes L AQ Silica Spherical, 7µm 0Å 00m /g % Monomeric Yes L C8 Silica Spherical,, 7, µm 0Å 00m /g % Monomeric No L7 C8e/c Silica Spherical,, 7, µm 0Å 00m /g % Monomeric Yes L7 C Silica Spherical µm 0Å 00m /g 6.% Monomeric Yes L6 Phe Silica Spherical µm 0Å 00m /g 9.% Monomeric Yes L CN Silica Spherical, µm 0Å 00m /g 7% Monomeric Yes L0 Amino (NH) Silica Spherical, µm 0Å 00m /g.% Polymeric No L8 Carbohydrate Silica Spherical µm 0Å 00m /g Monomeric CN-TCA Silica Spherical µm 0Å 00m /g 7% Monomeric Yes Petro-XP Silica Spherical µm 0Å 00m /g Monomeric Silica Silica Spherical,, 7, µm 0Å 00m /g n/a n/a n/a L Genesis C8 Reversed Phase Excellent peak symmetry Exceptional stability from ph to 0 Reduced need for mobile-phase modifiers Long column life Genesis C8 bonding is monomeric. A unique proprietary end-capping reagent, which is less prone to acid hydrolysis than trimethylsilane, provides freedom from residual silanols and enhanced stability under low-ph operating conditions. Genesis C8 columns also exhibit superior stability at alkaline ph. The permissible operating range is ph 0. Genesis AQ Reversed Phase Designed for separating hydrophilic and polar compounds Stable retention times in 00% aqueous mobile phases Rapid equilibration Unique reversed-phase selectivity The Genesis AQ adsorbent employs an optimum ratio of C8, short (non-tms) chains, and polar surface groups bonded to high-purity 0Å silica to allow rapid equilibration and provide consistent, reproducible chromatography with stable retention times in 00% aqueous eluents. 0 Min. Column: Flow Rate: Mobile Phase: Detector: Inertness Test. Uracil. Pyridine.,-Dimethylphenol. Methylbenzoate Genesis FM960E C8, µm,.6 x 0mm 0.8mL/min Isocratic, 0% ACN in water UV at nm Retention on Genesis AQ is greater for polar analytes but lower for non-polar compounds compared to Genesis C8. Uracil, which is typically an unretained peak on C8 columns, is well retained on the Genesis AQ adsorbent. Acids and bases exhibit good peak shapes. Although Genesis AQ excels with water-rich mobile phases, it can also be used in gradient and isocratic modes with a full spectrum of mobile phases. Nucleotide '-Monophosphates Min.. KNO. CMP. UMP. GMP. AMP Column: Genesis FM9E AQ, µm,.6 x 0mm Flow Rate:.0mL/min Mobile Phase: Isocratic, 0mM NHOAc, ph. Detector: UV at nm Column Temp: 0 C 6

42 Jones Genesis Columns Genesis C8(EC) Reversed Phase Excellent peak symmetry Exceptional stability from ph to 0 Reduced need for mobile-phase modifiers Long column life C8(EC) is double-bonded before fully endcapped using a unique proprietary end-capping reagent. It is less prone to acid hydrolysis than trimethylsilane, which provides freedom from residual silanols and enhanced stability under low-ph operating conditions. The bonding is monomeric. Genesis C8(EC) columns exhibit superior stability at alkaline ph. Genesis C8 Reversed Phase Non-endcapped Suitable for lower ph separations Genesis C8 bonding is monomeric. They are suitable for separations under lower ph conditions and may offer selectivity advantages for some samples. Genesis Phenyl Unique reversed-phase chemistry Improve the chromatography of polar aromatic, fatty acids, and basic pharmaceuticals Genesis Phenyl columns provide impressive peak symmetry for both acidic and basic compounds. Genesis Columns Genesis Guards Length: 0mm 0mm 00mm 0mm 00mm 0mm 0mm 0mm Genesis AQ µm.0mm i.d. FJ9E FJ09E FJ9E FJ9E FJ9-.mm i.d. FK9E FK9E FK09E FK9E FK09E FK9E FK9- FK9-.0mm i.d. FL9E FL9E FL09E FL9E FL9E FL9- FL9-.0mm i.d. FH9E FH9E FH09E FH9E FH9E FH9- FH9-.6mm i.d. FM9E FM9E FM09E FM9E FM09E FM9E Genesis C8 µm.0mm i.d. FJ96E FJ096E FJ96E FJ96E FJ96-.mm i.d. FK96E FK96E FK096E FK96E FK096E FK96E FK96- FK96-.0mm i.d. FL96E FL96E FL096E FL96E FL96E.0mm i.d. FH96E FH96E FH096E FH96E FH96E FH96- FH96-.6mm i.d. FM96E FM96E FM096E FM96E FM096E FM96E µm.0mm i.d. FJ960E FJ0960E FJ960E FJ960E FJ960-.mm i.d. FK960E FK960E FK0960E FK960E FK0960E FK960E FK960- FK960-.0mm i.d. FL960E FL960E FL0960E FL960E FL960E FL960- FL960-.0mm i.d. FH960E FH960E FH0960E FH960E FH960E FH960- FH960-.6mm i.d. FM960E FM960E FM0960E FM960E FM0960E FM960E Genesis C8 µm.0mm i.d. FJ968E FJ0968E FJ968E FJ968E FJ968-.mm i.d. FK968E FK968E FK0968E FK968E FL0968E FK968E FK968- FK968-.0mm i.d. FL968E FL968E FL0968E FL968E FL968E FL968- FL968-.0mm i.d. FH968E FH968E FH0968E FH968E FH968E FH968- FH968- µm.0mm i.d. FJ96E FJ096E FJ96E FJ96E FJ96-.mm i.d. FK96E FK96E FK096E FK96E FK096E FK96E FK96- FK96-.0mm i.d. FL96E FL96E FL096E FL96E FL96E FL96- FL96-.0mm i.d. FH96E FH96E FH096E FH96E FH96E FH96- FH96-.6mm i.d. FM96E FM96E FM096E FM96E FM096E FM96E Genesis C8(EC) µm.0mm i.d. FJ969E FJ0969E FJ969E FJ969E FJ969-.mm i.d. FK969E FK969E FK0969E FK969E FK0969E FK969E FK969- FK969-.0mm i.d. FL969E FL969E FL0969E FL969E FL969E FL969- FL969-.0mm i.d. FH969E FH969E FH0969E FH969E FH969E FH969- FH969-.6mm i.d. FM969E FM969E FM0969E FM969E FM0969E FM969E µm.0mm i.d. FJ96E FJ096E FJ96E FJ96E FJ96-.mm i.d. FK96E FK96E FK096E FK96E FK096E FK96E FK96- FK96-.0mm i.d. FL96E FL96E FL096E FL96E FL96E FL96- FL96-.0mm i.d. FH96E FH96E FH096E FH96E FH96E FH96- FH96-.6mm i.d. FM96E FM96E FM096E FM96E FM096E FM96E Genesis Phenyl µm.0mm i.d. FJ980E FJ0980E FJ980E FJ980E FJ980-.mm i.d. FK980E FK980E FK0980E FK980E FK0980E FK980E FK980- FK980-.0mm i.d. FH980E FH980E FH0980E FH980E FH980E FH980- FH980-.6mm i.d. FM980E FM980E FM0980E FM980E FM0980E FM980E FH980- FH980- NOTE: Genesis line is completed by additional phases such as Silica, Phenyl, Cyano, and Amino, for details please check online at. All Genesis guards listed are cartridges and require either a stand-alone holder or direct-connect holder for use. Guard cartridges are /pk. Guard Cartridge Holders for Genesis and Apex Guards 0 mm Stand-Alone 0 mm Stand-Alone 0 mm Direct-Connect 0 mm Direct-Connect.0 mm i.d. F9GPH. mm i.d. F9P F9P F9P.0,.0,.6 mm i.d. F9P F9P F9P F9P 6

43 Jones Apex Reversed-Phase Columns Economical Column Line with Maximum Reproducibility and Efficiency Conventional 00Å pore size spherical silica Uniform-sized particle Narrow particle distribution Controlled surface area Apex Phase Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code ODS Silica Spherical,, 0µm 00Å 70m /g 0% Polymeric Yes L ODS II Silica Spherical, µm 00Å 70m /g 0.% Monomeric Yes L C8 Silica Spherical, µm 00Å 70m /g 7% Monomeric No L7 C8(EC) Silica Spherical, µm 00Å 70m /g 7% Monomeric Yes L7 C Silica Spherical, µm 00Å 70m /g.% Monomeric Yes L Diol Silica Spherical, µm 00Å 70m/g.% Monomeric No Phe Silica Spherical, µm 00Å 70m /g % Monomeric Yes L CN Silica Spherical, µm 00Å 70m /g % Monomeric No L0 Amino (NH) Silica Spherical, µm 00Å 70m /g % Monomeric No L8 Amino II (NH) Silica Spherical, µm 00Å 70m /g % Monomeric No L8 Carbohydrate Silica Spherical µm 00Å 70m /g Monomeric Proprietary Silica Silica Spherical,, 0µm 00Å 70m /g n/a% n/a No L Apex HPLC Columns Packing Format i.d. x Length Part No. ODS, µm Analytical.6 x 0mm 09 ODS, µm Analytical.6 x 0mm 0970 Analytical.6 x 0mm 090 Analytical.6 x 0mm 0900 Analytical.6 x 0mm 090 ODS II, µm Analytical.6 x 00mm 099 Analytical.6 x 0mm 0 ODS II, µm Analytical.6 x 0mm 099 Analytical.6 x 00mm 99 Analytical.6 x 0mm 0970 Analytical.6 x 0mm 097 C8, µm Analytical.6 x 0mm Analytical.6 x 0mm 099 Analytical.6 x 0mm 097 C8(EC), µm Analytical.6 x 0mm Analytical.6 x 0mm 097 Diol, µm Analytical.6 x 0mm 07 Phenyl, µm Analytical.6 x 00mm 0997 Analytical.6 x 0mm 060 Analytical.6 x 0mm 099 Cyano, µm Analytical.6 x 0mm 0977 Cyano II, µm Analytical.6 x 0mm 099 Amino, µm Analytical.6 x 0mm Analytical.6 x 0mm 09 Amino II, µm Analytical.6 x 0mm 00 Silica, µm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm 00 Analytical.6 x 0mm 0 Apex Guard Columns Packings i.d. x Length Part No. ODS, µm x 0mm 070 ODS II, µm x 0mm 070 C8, µm x 0mm C8-EC, µm x 0mm Cyano, µm x 0mm 079 Amino, µm x 0mm 078 x 0mm Guard Cartridge Holder 0mm F9P 0mm F9P Column: Flow Rate: Mobile Phase: Apex ODS II, C8, µm,.6 x 0mm 0.8mL/min A: 0 mm NaOAc, 0.0% THF B: 0 mm NaOAc,.7% THF, % ACN Gradient: Time: 0 %B: 0 70 Column Temp: 0 C Detector: UV at 7nm 6 7 Xanthines in Plasma Min. more applications To view our complete searchable chromatogram database visit /chromdb/. 7-Methylxanthine. -Methylxanthine. -Methylxanthine.,-Methyluric Acid. Theobromine 6. Ethylxanthine 7. Methyluric Acid 8. Paraxanthine 9. Theophylline 0.,,7-Methyluric Acid. Ethyltheophylline. Caffeine 66

44 Alltech OPA Amino Acid Columns OPA-HS for high-speed with protein hydrolysates OPA-HR for high-resolution with physiological samples OPA-HS (high-speed) columns are ideal for the study of protein acid hydrolysates with very simple mobile-phase systems. OPA-HR (high-resolution) columns are ideal for applications that require more resolution than simple hydrolysates. OPA-HR gives baseline separation of most amino acids found in physiological fluids. 0 OPA-Derivatized Amino Acids Min. OPA Amino Acid HPLC Columns Packing Format i.d. x Length Part No. OPA-HS, µm Analytical.6 x 00mm 806 OPA-HR, µm Analytical.6 x 0mm 807 Derivatization Reagents Description Qty. Part No. OPA Crystals g Aspartic Acid. Glutamic Acid. Serine. Histidine. Glycine 6. Threonine 7. Arginine 8. Alanine 9. Tyrosine 0. Methionine. Valine. Phenylalanine. Isoleucine. Leucine. Lysine Column: OPA-HR, µm,.6 x 0mm (Part No. 807) Mobile Phase: A: Methanol B: Tetrahydrofuran C: 0mM Sodium Acetate buffer, ph. Gradient: Time: 0 0 %A: 6 %B: %C: 90 Flow Rate:.mL/min Detector: Fluorescence Alltech Catecholamine Columns For routine analysis of indoles and catecholamines Reliable and inexpensive With this column, easily resolve catecholamines under isocratic conditions at room temperature. A simple monochloroacetic acid mobile phase with varying amounts of sodium octyl sulfate and disodium EDTA controls the elution profile. Catecholamine HPLC Columns Packing Format i.d. x Length Part No. Catecholamine, µm Analytical.6 x 00mm 896 All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Catecholamine, µm.6 x 7.mm All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Alltech Nucleotide- Nucleoside Columns Determine both nucleotides and nucleosides with a single column With this column, separate seven common nucleosides or twelve common nucleotides using a buffer/methanol/tetrabutyl ammonium phosphate gradient. Nucleotide-Nucleoside HPLC Columns Packing Format i.d. x Length Part No. Nucleotide-Nucleoside, 7µm Analytical.6 x 0mm 6 All-Guard Cartridges* Packing i.d. x Length Qty. Part No. Nucleotide-Nucleoside, 7µm.6 x 7.mm All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. 70 more info For related products, see our Vydac line of Life Science columns on pages

45 Alltech Mixed-Mode Columns Separate hydrophobic and ionic species on a single column Control ionization with buffer adjustments instead of ion-pair reagents Anesthetics and Neutral Biphenyl. Lidocaine. Procaine Alltech Carbohydrate Cation Columns High efficiency separations using only water as the mobile phase Sulfonated polystyrene resin in the calcium form Column heating required 00% Pure Red Delicious Apple Juice (a :0 Dilution) 896. Unknown. Sucrose. Glucose. Fructose. Sorbitol Min. Column: Mixed-Mode C8/Cation, µm,.6 x 0mm Mobile Phase: 8780: mm KHPO, ph6.: Acetonitrile (0:0) 878: mm KHPO, ph6.: Acetonitrile (0:0) Flow Rate:.0mL/min Column Temp: 0 C Detector: UV at 0nm Changing the buffer concentration affects the capacity of the cationic components lidocaine and procaine, but does not affect the capacity of biphenyl, a neutral component. Mixed-Mode HPLC Columns Packing Format i.d. x Length Part No. Waters Fittings Part No. C8/Anion, 7µm Analytical.6 x 0mm 7 7 Analytical.6 x 0mm C8/Cation, µm Analytical.6 x 0mm Analytical.6 x 0mm C8/Cation, µm Analytical.6 x 0mm Analytical.6 x 0mm Carbohydrate Cation HPLC Columns Packing Format i.d. x Length Part No. Carbohydrate Cation, 0µm Analytical 6. x 00mm 7007 more info For Alltech Prevail Carbohydrate ES columns, see page 7. 0 Min. Column: Alltech Carbohydrate Cation, 0µm, 6. x 00mm (Part No. 7007) Mobile Phase: Water Flow Rate: 0.mL/min Column Temp: 90 C Detector: Alltech ELSD Carbohydrate Cation All-Guard Cartridges Packing i.d. x Length Qty. Part No. Carbohydrate Cation Guard Cartridges.6 x 7.mm 9609 All-Guard Cartridge Holder 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Mixed-Mode All-Guard Cartridges* Packing, µm i.d. x Length Qty. Part No. C8/Anion.6 x 7.mm 967 C8/Cation.6 x 7.mm 779 C8/Cation.6 x 7.mm 780 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. related product Need a column heater? See pages 6 and 7. related product Doing carbohydrate analysis? Alltech 00 ELSD is more sensitive than RI. See pages B 68

46 Vydac Venture Immunoaffinity Columns Repetitive-Use, Silica-Based Immunoaffinity Columns Venture immunoaffinity HPLC columns provide an easy and quick way to purify and concentrate analytes from complex food, environmental, and biological matrices for fast, accurate measurement Vydac Venture immunoaffinity columns use ICEtech (Inert Coating Enhancement Technology, U.S. Patent No. 6,80,966) silica processing to eliminate non-specific binding on the silica surface. This enables Venture columns to be the first affinity matrix to take advantage of silica s rigid porous structure, making direct coupling of the columns with other modes of chromatography, such as reversed-phase HPLC feasible. Benefits Features Rapid Analysis No laborious sample preparation procedures Fast binding kinetics Wide-pore silica support allows online coupling with HPLC for real time direct analysis Excellent Accuracy and Quantitative recovery of analyte from Precision IAC column High sensitivity due to high volume injection Precision comparable with conventional HPLC methods Highly Selective Specially treated wide-pore silica eliminates non-specific interactions Secondary separation reduces errors due to cross reactivity Highly selective antibodies toward target analyte Long Column Lifetime Analyze over 00 samples on a single column without column deterioration due to the high ligand stability of the Venture IAC columns Analysis of Difficult Matrices Venture IAC columns isolate, concentrate and purify targeted sample components from dilute and complex matrices in one step Significant Cost Reductions Costs reduced by as much as 90% Long column lifetime decreases material costs to less than $.00 per analysis Rapid analysis and automation reduces technician time Significant reduction in solvent use Less solvent waste disposal costs Automated Analysis HPLC format allows automation for high sample throughput Venture Immunoaffinity Columns Product Name Target Analyte i.d. and length Part No. Mycotoxins Analysis Venture AF Aflatoxin. x 0mm 0 Venture OT Ochratoxin A. x 0mm 9 Food Nutrient Analysis Venture FA Folic Acid. x 0mm 9 Venture B Vitamin B. x 0mm 00 Venture BF Folic Acid and Vitamin B. x 0mm 9 Acrylamide Analysis Venture AC Acrylamide. x 0mm 9 Lactoferrin Analysis Venture LTF Lactoferrin. x 0mm 000 Endocrine Disruptors Analysis Venture EE 7α-Ethynyl Estradiol. x 0mm 00 Venture E 7β-Estradiol. x 0mm 00 Venture E Estrone. x 0mm 00 Venture BPA Bisphenol A. x 0mm 006 Pollutants Analysis Venture CPA Chlorophenoxy Acetic Acid. x 0mm 007 Herbicides Venture PVH Phenylurea Herbicides. x 0mm 008 Venture OPP Organophosphorous. x 0mm 00 Pesticides Venture VCZ Vinclozolin Fungicide. x 0mm 0 Steroid Hormones Analysis Venture TT Testosterone. x 0mm 00 Venture NT Nortestosterone. x 0mm 00 Vydac Denali C8 Reversed Phase Denali columns contain a new-generation silica-based 0Å monomeric C8 adsorbent based on the same bonding technology employed in Vydac Everest 00Å columns. This new technology improves C8 surface coverage and deactivates residual silanols. Previously, the best monomeric C8 chemistries had carbon coverage in the.8 to.6µmol/m range. Denali C8 has coverage in excess of µmol/m and approximates the theoretical limit based on surface area. 70 Vydac Denali C8 Reversed-Phase 8DE C8 Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge µm.mm 8DE0 8DE0.6mm 8DE0 8DE0 8GKDE 8GDDE µm.0mm 8DE0 8DE0 8DE 8DE 8GKDE 8GDDE.mm 8DE0 8DE0 8DE 8DE 8GKDE 8GDDE.6mm 8DE0 8DE0 8DE 8DE 8GKDE 8GDDE NOTE: Additional column diameters and lengths are available on request. Please contact Grace Davison Discovery Sciences to discuss your requirements. A guard kit includes a holder and one guard cartridge. Guard cartirdge units include two guard cartridges. Titanium frits are standard in column diameters.6mm and smaller. 69

47 Vydac 0TP and 0TP C8 Specialty Reversed Phases for PAH and Vitamins 0TP: The standard for PAH analysis in all types of environmental samples. Vydac 0TP columns separate the EPA 6 priority pollutants in less than 0 minutes. 0TP0: Rapid analysis to separate the 6 priority pollutant PAHs in under 0 minutes 0TP and 0TP: For the analysis of derivatized PAHs Vydac 0TP and 0TP columns were developed specifically for the separation and quantification of PAHs required by environmental regulations, current and future. Beyond the 6 EPA priority pollutant PAHs, Vydac PAH columns are used to separate many other PAHs, such as methylated naphthalenes. Vydac 0TP and 0TP C8 Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code 0TP C8 Silica Spheroidal, 7, 0, 0, 0µm 00Å 70 90m /g 8% Polymeric No L 0TP C8 Silica Spheroidal,, 0µm 00Å 60 90m /g 9% Polymeric No L Vydac 0TP Vydac 0TP HPLC columns have long been the standard for the analysis of PAHs in water, air, soil, automotive exhaust, and food. They were used to establish standard reference materials, measure air quality, measure PAHs in sediments, quantify PAHs in food, and study high molecular weight PAHs. They have also been used in the study of shape selectivity of reversed-phase materials. Priority Pollutants PAHs In Accordance with EPA Methods 0, 0., 60, and 80. Naphthalene. Acenaphthylene. Acenaphthene. Fluorene. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene 9. Benz[a]anthracene 0. Chrysene. Benzo[b]fluoranthene. Benzo[k]fluoranthene. Benzo[a]pyrene. Dibenz[ah]anthracene. Benzo[ghi]perylene 6. Indeno[,,-cd]pyrene 6 PAHs Beyond the EPA Priority Pollutants. Naphthalene. Acenaphthylene. -Methylnaphthalene. -Methylnaphthalene. Acenaphthene 6. Fluorene 7. Phenanthrene 8. Anthracene 9. Fluoranthene 0. Pyrene. Benzo[c]phenanthrene. Cyclopenta[cd]pyrene 8. Benz[a]anthracene. Chrysene. Benzo[b]naptho[,-d]thiopen 6. 7,-Dimethylbenz[a]anthracene 7. Benzo[e]pyrene 8. Benzo[b]fluoranthene 9. Benzo[k]fluoranthene 0. Benzo[a]pyrene. Dibenz[ah]anthracene. Benzo[ghi]perylene. Indeno[,,-cd]pyrene Min Min. Column: Flow Rate: Mobile Phase: Gradient: Detector: Vydac C8, µm,.6 x 0mm (Part No. 0TP).mL/min A: Water B: ACN Hold 0% B for min, then 0 to 00% B in 7min UV at nm Column: Vydac C8, µm,.6 x 0mm (Part No. 0TP) Flow Rate:.0mL/min Mobile Phase: A: Water B: ACN Gradient: 0 to 00% B over 0min Column Temp: 0 C Detector: UV at nm more applications To view our complete searchable chromatogram database visit /chromdb/ 70

48 Vydac 0TP (continued) Vitamins Vydac 0TP columns also have application in the analysis of carotenoids, retinoids, and vitamins. Vydac 0TP Built on top of the success of 0TP, 0TP columns are ideal for derivatized PAH samples or high throughput PAH analyses. Retinol, a-tocopherol, and b-carotene in Serum. All-trans-retinol (vitamin A). Tocol. g-tocopherol. a-tocopherol (vitamin E). Lutein 6. Zeaxanthin 7. Cryptoxanthin 8. a-carotene 9. All-trans-b-carotene 0. Cis-b-carotene nm 9 nm 0 nm Min.. Naphthalene. Acenaphthylene. Acenaphthene. Fluorene. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene Rapid Analysis of Polyaromatic Hydrocarbons Benz[a]anthracene 0. Chrysene. Benzo[b]fluoranthene. Benzo[k]fluoranthene. Benzo[a]pyrene. Benzo[ghi]perylene. Dibenz[ah]anthracene 6. Indeno[,,-cd]pyrene 6 Column: Flow Rate: Mobile Phase: Gradient: Detector: Vydac 0TP C8, µm,.6 x 0mm (Part No. 0TP).mL/min A: :7:0 Buffer/Methanol/n-Butanol B: :88:0 Buffer/Methanol/n-Butanol Buffer = Aqueous 0.0 M NHOAc, ph. 00% A for min, then 0 to 00% B over min, then hold 00% B UV, programmed wavelengths Min. Column: Vydac 0TP C8, µm,.6 x 0mm (Part No. 0TP0) Flow Rate:.0mL/min Mobile Phase: A: Water B: ACN Gradient: 0 to 9% B in 8min Column Temp: 0 C Detector: UV at nm Chromatogram reproduced with author s permission. From W.A. MacCrehan and E. Schonberger, Clin. Chem., (9), 8-9 (987). 0TP C8 Analytical Columns Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge µm.0mm 0TP0 0TP 0TP 0GKT 0GDT.mm 0TP0 0TP0 0TP 0TP 0GKT 0GDT.mm 0TP 0TP 0GKT 0GDT.6mm 0TP0 0TP0 0TP 0TP 0GKT 0GDT 0µm.6mm 0TP0 0TP0 0GK0T 0GD0T A guard kit includes a holder and one guard cartridge. Guard cartridge units include two guard cartridges. 0TP High-Carbon-Load C8 Analytical Columns Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge µm.mm 0TP.6mm 0TP0 0TP0 µm.6mm 0TP 0TP 0GKT 0GDT 0µm.6mm 0TP00 0GK0T 0GD0T A guard kit includes a holder and one guard cartridge. Guard cartridge units include two guard cartridges. 7

49 Astec Amino and Reversed-Phase Columns Durable vinyl alcohol copolymer Stable from ph Stable and reproducible performance Choose Astec amino columns for separating mono- and oligosaccharides. Astec s polymer base makes these columns much more stable than silica-based amino columns. Choose Astec reversed-phase columns for amines and high recoveries of proteins and peptides. Because there are no silanols, elution order is always based on hydrophobicity of the analytes rather than polar interactions with the base material. High recovery of proteins and peptides is typical even at low sample loads. Mono-, Di-, and Trisaccharides Rhamnose. Fructose. Glucose. Sucrose. Maltose 6. Raffinose Astec Cyclobond Columns For Chiral Separations Versatile chiral selectors High-purity µm spherical silica Cyclodextrin Phases Designation Substituent I 000 β-cyclodextrin I 000 Ac β-cyclodextrin Acetylate I 000 RSP β-cyclodextrin R,S-Hydroxypropyl Ether I 000 DMP β-cyclodextrin,-dimethylphenyl Carbamate II γ-cyclodextrin Cyclobond I 000 has the broadest applicability, and is ideal for small analytes in pharmaceutical, chemical, and environmental applications. Cyclobond 000 Ac is ideal for aromatic alcohols or amines that are chiral on the alpha or beta carbon. Cyclobond I 000 RSP is a general-purpose chiral stationary phase. It can separate non-aromatic structures such as t-boc amino acids. Cyclobond DMP shows good selectivity when the chiral centers are part of a ring structure or on the alpha carbon. This phase is ideal for derivatized amines such as amphetamine ACQ. Cyclobond II is useful for isomeric compounds based on anthracene, chrysene, and pyrene ring structures. Some applications include steroids, porphyrins, and FMOC amino acids Min. Column: Astec Amino, µm,.6 x 0mm (Part No. 60) Mobile Phase: Acetonitrile:Water (7:) Flow Rate:.0mL/min Detector: RI Astec Amino and Reversed-Phase 00Å Columns Packing Format i.d. x Length Part No. C8, µm Analytical.6 x 0mm 600 Analytical.6 x 0mm 600 C8, µm Analytical.6 x 0mm 6006 Analytical.6 x 0mm 6007 C, µm Analytical.6 x 0mm 6008 Analytical.6 x 0mm 6009 Amino, µm Analytical.6 x 0mm 60 Astec Amino and Reversed-Phase Guard Columns Packing i.d. x Length Qty. Part No. C8, µm.6 x 0mm ea 880 C8, µm.6 x 0mm ea 88 C, µm.6 x 0mm ea 88 Amino, µm.6 x 0mm ea 887 Astec Cyclobond Columns Packing Format i.d. x Length Part No. I 000, µm Analytical.6 x 00mm 000 Analytical.6 x 0mm 00 I 000 Ac, µm Analytical.6 x 0mm 0 I 000 RSP, µm Analytical.6 x 0mm I 000 DMP, µm Analytical.6 x 0mm II Analytical.6 x 00mm 000 Analytical.6 x 0mm 00 Astec Cyclobond Guard Cartridges* Packing, i.d. x Length Qty. Part No. I 000, µm.0 x 0mm ea 00 I 000 Ac, µm.0 x 0mm ea 00 I 000 RSP, µm.0 x 0mm ea 00 I 000 DMP, µm.0 x 0mm ea 008 II.0 x 0mm ea 009 Astec Guard Cartridge Holder ea 0 *Guard holder required. 7

50 Astec Chirobiotic HPLC Columns For Multi-Mode Chiral Separations Bonded macrocyclic glycopeptide phases Three complimentary selectivities High-purity µm spherical silica Chirobiotic columns demonstrate a broad selectivity in reversed phase, normal phase, and polar organic modes. This gives Chirobiotic columns the ability to separate a greater variety of chiral analytes than columns that can only operate in one mode. Chirobiotic V Chirobiotic V Specifications Ligand: Vancomycin Ideal For: Neutral molecules, amides, acids, esters, cyclic amines Chirobiotic V has a selectivity similar to glycoprotein phases while also being stable from 0 00% organic modifiers. New Chirobiotic V has enhanced selectivity and capacity in the polar organic mode, and increased capacity. Naproxen 8769 CH CHCOH O H CO Min. Column: Chirobiotic V,.6 x 0mm (Part No. 0) Mobile Phase: THF:0.% Triethylammonium Acetate, ph 7 (0:90) Flow Rate:.0mL/min Detector: UV at nm Phenylalanine 8770 O Chirobiotic T Chirobiotic T Specifications Ligand: Teicoplanin Ideal For: Underivatized amino acids, n-derivatized amino acids, carboxylic acids, phenols, neutral aromatics, cyclic aromatics with aliphatic amines Chirobiotic T is an excellent alternative to crown ether and ligand exchange for amino acids and hydroxy acids. New Chirobiotic T has enhanced selectivity and capacity in the polar organic mode. Chirobiotic R Chirobiotic R Specifications Ligand: Ristocetin A Ideal For: Anionic molecules, di- and tri-peptides, α-hydroxy acids, substituted aliphatic acids, aromatic esters, chiral alcohols, secondary and tertiary amines Chirobiotic R is the largest and most complex of the Chirobiotic ligands. Sugar moieties, a peptide chain, and additional ionizable groups give this structure the complexity and diversity to separate a wide variety of analytes Min. NH Column: Chirobiotic T,.6 x 0mm (Part No. 0) Mobile Phase: Ethanol:Water (80:0) Flow Rate:.0mL/min Detector: UV at 0nm Astec Chirobiotic Columns Packing Format i.d. x Length Part No. Chirobiotic V Analytical.6 x 0mm 6 Analytical.6 x 0mm 0 Analytical.6 x 0mm 0 Chirobiotic V Analytical.6 x 00mm A0 Analytical.6 x 0mm A0 Analytical.6 x 0mm A0 Prep 0 x 0mm A0 Prep. x 0mm A0 Chirobiotic T Analytical.6 x 0mm 9 Analytical.6 x 0mm 0 Analytical.6 x 0mm 0 Chirobiotic T Analytical.6 x 00mm A60 Analytical.6 x 0mm A60 Analytical.6 x 0mm A60 Prep 0 x 0mm A60 Prep. x 0mm A60 Chirobiotic R Analytical.6 x 0mm Analytical.6 x 0mm 6 Analytical.6 x 0mm 8 Astec Chirobiotic Guard Cartridges* Packing i.d. x Length Qty. Part No. Chirobiotic V, µm.0 x 0mm ea 09 Chirobiotic T, µm.0 x 0mm ea Chirobiotic R, µm.0 x 0mm ea Astec Guard Cartridge Holder ea 0 *Guard holder required. OH 7

51 hplc columns small and large molecule 7 Brownlee Columns Economical MPLC Cartridge System with Reusable Hardware Finger-tight modular design Reusable end assemblies with interchangeable holders Guards fit conventional columns or MPLC hardware The Brownlee MPLC Cartridge system is convenient and easy to use. Decide which column length you prefer, and purchase the hardware and the disposable packed cartridges separately. Cartridge Hardware Brownlee MPLC Cartridge Hardware Photo Description Part No. MPLC Cartridge Holders 0mm 000 MPLC Cartridge Accessories 067B 067A Brownlee MPLC NewGuard Hardware Photo Description Part No. NewGuard Cartridge 060 Holders NewGuard Holder 067E for use with Conventional Columns 0mm NewGuard 00 System Holder for Direct Coupling of 0mm 067F Analytical Cartridge with a NewGuard Cartridge NewGuard End Assembly mm 00 0mm 000 MPLC Cartridge Union 000 MPLC Cartridge End C Assembly MPLC Cartridge Holder Bodies 0mm D 00mm 000 0mm 00 Universal MPLC Cartridge Holder Kit Includes: 0-, 00-, and 0mm MPLC Cartridge Holder Bodies (E), MPLC Cartridge Union (C), End Assemblies (D), and NewGuard End Assembly (H) Brownlee MPLC Cartridge Hardware Seals Description Analytical Holder Seal Replacement Kit Includes Tool and Seals Analytical Seals, /pk Seal Replacement Tool Required NewGuard System 067G 00 Part No Brownlee NewGuard Cartridges* Description Qty. Part No. RP-8 00 RP-8 00 Cyano 008 Amino 007 Silica 0 Anion (AX-00) 009 *NewGuard cartridge holder required (Part No. 060). Spheri- and Spheri-0 Cartridges Rugged and reproducible Use with MPLC cartridge hardware 80Å, spherical silica RP-8 and RP-8 phases are monomeric for optimum mass transfer kinetics and high efficiency. The ODS phase is polymeric and more suitable for acidic mobile phases. Brownlee Spheri- Cartridges* Packing i.d. x Length Qty. Part No. ODS (C8), µm. x 0mm 06. x 00mm ea 06. x 0mm ea 06.6 x 0mm x 00mm ea 06.6 x 0mm ea 06 RP-8, µm. x 0mm 0. x 00mm ea 0. x 0mm ea 06.6 x 00mm ea 0.6 x 0mm ea 0 RP-8, µm. x 0mm 08. x 00mm ea 00. x 0mm ea 0.6 x 0mm 07.6 x 00mm ea 09.6 x 0mm ea 0 Cyano, µm.6 x 0mm 08.6 x 00mm ea 08.6 x 0mm ea 087 Amino, µm.6 x 0mm 77.6 x 00mm ea 79.6 x 0mm ea 8 Silica, µm. x 0mm 9.6 x 0mm 9.6 x 00mm ea 9.6 x 0mm ea 97 *Brownlee MPLC cartridge holder required. Brownlee Spheri-0 Cartridges* Packing i.d. x Length Qty. Part No. RP-8, 0µm.6 x 0mm 0.6 x 00mm ea 0 RP-8, 0µm.6 x 0mm 00.6 x 00mm ea 0 Silica, 0µm.6 x 0mm 86 *Brownlee MPLC cartridge holder required. Aquapore DEAE WAX Cartridges Large pore for macromolecules Choose AX-00 with its DEAE functional group for weak anion exchange applications with proteins and peptides. Brownlee Aquapore Cartridges* Packing i.d. x Length Qty. Part No. AX-00, 7µm.6 x 0mm.6 x 00mm ea *Brownlee MPLC cartridge holder required.

52 Jordi Columns Normal- and Reversed-Phase Rugged use at high temperatures, from ph 0, with any solvent Silanol Free analyze polar compounds without silanol effects Jordi normal-phase and reversed-phase columns are made from highly pure µm DVB, for extremely durable and efficient phases. Jordi Specifications Phase Particle Size Pore Size Description RP-DVB µm 00Å, 000Å Reversed-phase, not bonded C8-DVB µm 00Å Reversed-phase, C8 bonded Polyamine-DVB µm 00Å Normal-phase RP-DVB RP-DVB is a reversed-phase non-polar packing with a high degree of aromatic character. SM-00, with a 00Å pore, is recommended for most applications. LM-000, with a 000Å pore, is ideal for molecules 00,000 Daltons or larger. C8-DVB C8-DVB has C8 chains bonded to the DVB base. These columns are more hydrophobic than traditional C8 columns and have different selectivities due to the absence of silanols. Polyamine-DVB Polyamine-DVB is a normal-phase column for simple sugar and polysaccharide applications. The DVB-based packing provides a non-reactive amine surface that lasts longer and equilibrates faster than traditional silica-based amine columns. Polyamine-DVB columns can be used with aqueous mobile phases, even M NaOH. Ideal for carbohydrate analyses requiring sharp peaks and short run times. Jordi HPLC Columns Packing Format i.d. x Length Part No. RP-DVB (SM-00Å), µm Analytical.6 x 0mm 0000 Analytical.6 x 0mm 000 RP-DVB (LM-000Å), µm Analytical.6 x 0mm 0080 Analytical.6 x 0mm 008 C8-DVB (00Å), µm Analytical.6 x 0mm 800 Analytical.6 x 0mm 80 Polyamine-DVB (00Å), µm Analytical.6 x 0mm Jordi All-Guard Cartridges* Packing i.d. x Length Qty. Part No. RP-DVB (SM-00Å), µm.6 x 7.mm 009 RP-DVB (LM-000Å), µm.6 x 7.mm 0096 C8-DVB (00Å), µm.6 x 7.mm 0060 Polyamine-DVB (00Å), µm.6 x 7.mm 006 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. related product Looking for column heaters? See pages 6 and 7. GPC Columns Organic GPC Rugged highly crosslinked DVB for broad temperature stability and solvent compatibility Powerful high pore volume for greater resolution from a single column Jordi Organic and Aqueous GPC Specifications Pore Size Pressure Limit MW Range (Daltons) 00Å 8000psig Å 8000psig 00 0,000 0Å 8000psig 00,000 0Å 000psig 00,000,000 0Å 000psig 0,000 0,000,000 Mixed Bed 000psig 00 >0,000,000 Jordi HPLC columns, offer unparalleled resistance to shrinking and swelling, as well as better temperature and solvent compatibility than traditional ps-dvb phases. The high pore volume speeds up your analyses. A single 0 x 00mm Jordi column replaces three 7.8 x 00mm ps-dvb columns and separates the same sample in 0% less time. Aqueous GPC Glucose-DVB packings are rugged, hydrophilic packings for separating polar compounds. Glucose units are bonded to the DVB base to yield a hydrophilic surface. Glucose-DVB columns equilibrate faster than silica-based columns. Jordi Organic GPC Columns* Pore Size Format i.d. x Length Part. No. 00Å, µm Semi Prep 7.8 x 00mm 00 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm 006 Prep 0 x 00mm Å, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm 007 0Å, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm 0077 Mixed-Bed Linear, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm 008 Prep 0 x 00mm 008 *Jordi GPC columns are constructed from Type 6 stainless steel. All tubing connections are /6" female. Columns are shipped in Chloroform unless otherwise requested when placing order. Jordi Aqueous GPC Columns Pore Size Format i.d. x Length Part No. 00Å, µm Semi Prep 7.8 x 00mm 07 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 08 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 09 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 00 Prep 0 x 0mm Å, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm Mixed-Bed Linear, µm Semi Prep 7.8 x 00mm 0 Prep 0 x 0mm 0078 Prep 0 x 00mm 0078 hplc columns small and large molecule 69 7

53 Merck LiChrosorb and LiChrospher Columns LiChrosorb irregular silica for larger surface areas LiChrospher spherical silica for higher stability and reproducibility All Merck columns are packed by Grace to our highest QC standards. Custom columns are available. LiChrosorb HPLC Columns Packing Format i.d. x Length Part No. RP-8, µm Solvent Reducer.0 x 0mm 8 Solvent Reducer.0 x 0mm 8 Analytical.6 x 0mm 868 RP Select B, µm Analytical.6 x 0mm 8700 Analytical.6 x 0mm 868 RP-8, µm Solvent Reducer.0 x 0mm 8 Solvent Reducer.0 x 0mm 86 Analytical.6 x 0mm 869 Amino, µm Analytical.6 x 0mm 887 Si-60, 0µm Analytical.6 x 0mm 869 Si-00, 0µm Analytical.6 x 0mm 870 Melatonin Min. Column: LiChrospher RP-8, µm,.6 x 0mm (Part No. 869) Mobile Phase: mm KHPO, ph.7:meoh (60:0) Flow Rate:.0mL/min Detector: UV at nm 90. Melatonin Alltech All-Guard System change cartridges in seconds without tools. LiChrospher HPLC Columns Packing Format i.d. x Length Part No. RP-8, µm Analytical.6 x 0mm 8688 RP-8, µm Analytical.6 x 0mm 869 Analytical.6 x 0mm 867 RP-8 EC, µm Analytical.6 x 0mm 8686 RP-8 EC, µm Analytical.6 x 0mm 869 Analytical.6 x 0mm 867 RP-8, µm Analytical.6 x 0mm 8696 Analytical.6 x 0mm 867 RP-8 EC, µm Analytical.6 x 0mm 8697 Analytical.6 x 0mm 8678 RP-Select B, µm Analytical.6 x 0mm 86 Analytical.6 x 0mm 86 Amino, µm Analytical.6 x 0mm 869 Analytical.6 x 0mm 8670 Diol-00, µm Analytical.6 x 0mm 869 Analytical.6 x 0mm 8666 Si-60, µm Analytical.6 x 0mm 8699 Analytical.6 x 0mm LiChrosorb All-Guard Cartridges* Packing i.d. x Length Qty. Part No. RP-8, µm.0 x 7.mm x 7.mm 967 RP-Select B, µm.6 x 7.mm 968 RP-8, µm.0 x 7.mm x 7.mm 967 Amino, µm.6 x 7.mm 967 Silica, µm.6 x 7.mm 960 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. LiChrospher All-Guard Cartridges* Packing i.d. x Length Qty. Part No. RP-8, µm.6 x 7.mm 968 RP-8 Endcapped, µm.6 x 7.mm 968 RP-8, µm.6 x 7.mm 9689 RP-8 Endcapped, µm.6 x 7.mm 9687 RP Select B, µm.6 x 7.mm 96 Amino, µm.6 x 7.mm 969 Diol, µm.6 x 7.mm 969 Silica, µm.6 x 7.mm 960 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. 76

54 Macherey-Nagel Nucleosil Columns 00Å, high-purity silica Narrow particle size distribution A choice of phases for any application All Nucleosil columns are packed by Grace to our highest QC standards. C8 AB is polymerically bonded specifically for reversedphase chromatography of acidic and basic compounds. It has a % carbon load for strong retention. 0 SRM Min. Column: Nucleosil C8 AB, µm,.6 x 0mm (Part No. 6) Mobile Phase: (8:) Acetonitrile:Water Flow Rate:.0mL/min Detector: UV at nm Column Temp: C 90. Acetone. Phenanthroanthrene.,:,:,6:7, 8-Tetrabenzonaphthalene. Benzo[a]pyrene Nucleosil HPLC Columns Packing Format i.d. x Length Part No. Waters Fittings Part No. C8, µm Analytical.6 x 00mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm C8 AB, µm Analytical.6 x 0mm 6 Analytical.6 x 0mm 67 C8, µm Analytical.6 x 0mm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm Phenyl, 7µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, 0µm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Analytical.6 x 0mm Amino, 0µm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm Silica, 0µm Analytical.6 x 0mm SA, µm Analytical.6 x 0mm Analytical.6 x 0mm SA, 0µm Analytical.6 x 0mm SB, µm Analytical.6 x 0mm Analytical.6 x 0mm SB, 0µm Analytical.6 x 0mm Opium Alkaloids Alltech All-Guard System change cartridges in seconds without tools Min. 6 Column: Nucleosil CN, 0µm,.6 x 0mm (Part No. 898) Mobile Phase: 80%mM Ammonium Acetate ph.8 0% Acetonitrile 0% Dioxane Flow Rate:.mL/min Detector: UV at 0nm Column Temp: Ambient. Morphine. Codeine. Cryptopine. Thebaine. Narcotine 6. Papaverine Nucleosil All-Guard Cartridges* Packing i.d. x Length Qty. Part No. C8, µm.6 x 7.mm 960 C8, µm.6 x 7.mm 96 Phenyl, µm.6 x 7.mm 96 Cyano, µm.6 x 7.mm 96 Amino, µm.6 x 7.mm 96 Silica, µm.6 x 7.mm 96 SA, µm.6 x 7.mm 890 SB, µm.6 x 7.mm 89 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. more info For more HPLC Applications, see pages more applications To view our complete searchable chromatogram database visit /chromdb/ 77

55 Whatman Partisil Columns All Whatman Partisil columns are packed by Grace to our highest QC standards. Partisil columns use 8Å, irregular silica with a surface area of 0m /g. Partisil PAC is a polar amino cyano phase with secondary amino groups for thermal and chemical stability. It separates primarily by reversed-phase or weak anion exchange. Partisil HPLC Columns Packing Format i.d. x Length Part No. Waters Fittings Part No. ODS, 0µm Analytical.6 x 0mm ODS-, µm Analytical.6 x 0mm 7 7 ODS-, 0µm Analytical.6 x 0mm 8 8 ODS-, µm Analytical.6 x 0mm ODS-, 0µm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm C8, 0µm Analytical.6 x 0mm PAC, µm Analytical.6 x 0mm PAC, 0µm Analytical.6 x 0mm Silica, µm Analytical.6 x 0mm Silica, 0µm Analytical.6 x 0mm SAX, µm Analytical.6 x 0mm SAX, 0µm Analytical.6 x 0mm SCX, 0µm Analytical.6 x 0mm Partisil All-Guard Cartridges* Packing i.d. x Length Qty. Part No. ODS, µm.6 x 7.mm 96 ODS-, µm.6 x 7.mm 96 ODS-, µm.6 x 7.mm 96 C8, µm.6 x 7.mm 96 PAC, µm.6 x 7.mm 966 Silica, µm.6 x 7.mm 967 SAX, µm.6 x 7.mm 969 SCX, µm.6 x 7.mm 968 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. Whatman Partisphere WVS Columns Whatman s Partisphere WVS columns have built-in void compensators that help you extend the column lifetime. Use the piston-like plungers at each end of the column to compress the bed and seal any voids that develop over time. Choose your column from the table below, and purchase reusable WVS endfittings separately. Partisphere WVS HPLC Cartridges* Packing Format i.d. x Length Part No. C8, µm Analytical.6 x mm 600 C8, µm Analytical.6 x mm 600 PAC, µm Analytical.6 x mm 6008 Silica, µm Analytical.6 x mm 600 SAX, µm Analytical.6 x mm 600 SCX, µm Analytical.6 x mm 6007 WVS Reusable Endfittings, /pk 6000 *Reusable endfittings required. Partisphere WVS Guard Cartridges* Packing i.d. x Length Qty. Part No. C8/C8, µm x 7mm 6000 PAC, µm x 7mm Silica, µm x 7mm 6000 SAX, µm x 7mm 6000 SCX, µm x 7mm Guard Cartridge Holder 9 x 0mm ea 600 *Guard holder required. more info For HPLC Applications, see pages 9. related products Need HPLC tubing? See pages 8 9. technical assistance Contact Tech Support: Phone: (North America) contact.alltech@grace.com Online: related products Looking for high-pressure fittings? See pages

56 Polymer Labs Columns Outstanding Durability and Stability Ideal for elevated temperature and pressure applications Stable from ph PLRP-S is a polystyrene divinylbenzene copolymer designed specifically for reversed-phase HPLC. PLRP-S columns are extremely stable, performing over a wide ph range and at elevated temperatures and pressures. PLRP-S Choice of Pore Sizes: 00Å for small molecules, peptides, and nucleotides 00Å for polypeptides and globular proteins 000Å for fibrous proteins 000Å for the analysis of very large biomolecules or high-speed/high-resolution separations PLRP-S HPLC Specifications Base Material Polystyrene Divinylbenzene Organic Modifier 0 to 00% Particle Size,, 8, 0µm Pore Size 00, 00, 000, 000Å ph Range Max. Temp. 80ºC Max. Pressure 000psig Max. Flow Rate ml/min Min. Column: Mobile Phase: Gradient: Flow Rate: Detector: Enkephalins PLRP-S 00Å, µm,.6 x 0mm (Part No. -00) A: 0.% TFA, % IPA in Water B: 0.% TFA, % IPA in ACN Linear 8% B in min.0ml/min UV at 0nm 7. Methionine Enkephalin Sulphoxide. Methionine Enkephalin Lysine. Methionine Enkephalin. D-ALA Methionine Enkephalin. D-TRP Methionine Enkephalin PLRP-S HPLC Columns Packing Format i.d. x Length Part No. 00Å, µm Microbore. x 0mm 9-00 Microbore. x 0mm 9-00 Analytical.6 x 0mm -00 Analytical.6 x 0mm Å, µm Microbore. x 0mm 9-00 Microbore. x 0mm 9-00 Microbore. x 0mm 9-00 Analytical.6 x 0mm -00 Analytical.6 x 0mm -00 Analytical.6 x 0mm Å, 8µm Analytical.6 x 0mm -800 Analytical.6 x 0mm Å, µm Microbore. x 0mm 9-0 Microbore. x 0mm 9-0 Analytical.6 x 0mm -0 Analytical.6 x 0mm -0 00Å, µm Microbore. x 0mm 9-0 Microbore. x 0mm 9-0 Microbore. x 0mm 9-0 Analytical.6 x 0mm -0 Analytical.6 x 0mm -0 Analytical.6 x 0mm -0 00Å, 8µm Analytical.6 x 0mm -80 Analytical.6 x 0mm Å, µm Microbore. x 0mm 9-0 Analytical.6 x 0mm Å, 8µm Microbore. x 0mm 9-80 Microbore. x 0mm 9-80 Analytical.6 x 0mm -80 Analytical.6 x 0mm -80 Analytical.6 x 0mm Å, µm Microbore. x 0mm 9-0 Analytical.6 x 0mm Å, 8µm Microbore. x 0mm 9-80 Microbore. x 0mm 9-80 Analytical.6 x 0mm -80 Analytical.6 x 0mm -80 PLRP-S Guard Columns Packing i.d. x Length Qty. Part No. PLRP-S, µm x mm -0 Guard Column Holder ea more info For more large molecule columns, see pages related product Looking for column heaters? See page 6 7. hplc columns small and large molecule 69 79

57 hplc columns small and large molecule Shodex Columns Aqueous Size Exclusion High efficiency and high mass recovery Separate biopolymers by their effective size in solution Stable sugar GFC columns Shodex Ohpak and Sugar columns are designed for highresolution separations of water-soluble compounds such as organics, inorganics, oligomers, and polymers. Three different gel filtration size exclusion materials are available. Shodex Sugar columns use a sulfonated gel with a sodium counter ion for separation of mono, di, oligo, and polysaccharides, starches, and celluloses. Proteins by GFC 96. Glutamate Dehydrogenase (90k). Lactate Dehydrogenase (k). Enolase (67k). Adenylate Kinase (k). Cytochrome (.k) Aqueous Size Exclusion Columns Packing Series Description Applications OHpak SB-800 Polyhydroxymethacrylate General purpose GFC of water-soluble polymers, proteins, and enzymes Protein KW-800 Porous Silica Gel GFC of proteins, glycoproteins, and peptides Sugar KS-800 Sulfonated PS Gel (Na+ counter ion) 6 Separation of mono, di, oligo, and polysaccharides, starches, and celluloses Pullulan, Saccharides, and Ethanol 96. Pullulan (6.7k). Pullulan (.k). Raffinose. Sucrose. Fructose 6. Ethanol 0 0 Min Min. Column: Shodex Protein KW-80, 8.0 x 00mm (Part No. 79) Mobile Phase: 0mM Phosphate buffer + 0.M NaCl, ph 7.0 Flow Rate:.0mL/min Detector: ELSD Column Temp: 8 C Column: Shodex Sugar KS-80, 8.0 x 00mm (Part No. 7) Mobile Phase: Water Flow Rate:.0mL/min Detector: RI Column Temp: 80 C Shodex Size Exclusion HPLC Columns Particle Size Format Packing MW Range i.d. x Length Part No. OHpak 8µm Analytical Q x 00mm 77 8µm Analytical SB-80 HQ x 00mm 70 6µm Analytical SB-80. HQ 0 0, x 00mm 7 6µm Analytical SB-80 HQ 0 00, x 00mm 7 0µm Analytical SB-80 HQ 00,000, x 00mm 7 µm Analytical SB-80 HQ 00,000, x 00mm 7 µm Analytical SB-806M HQ 00 0,000, x 00mm 76 Protein µm Analytical KW , x 00mm 78 µm Analytical KW , x 00mm 79 7µm Analytical KW , x 00mm 70 Sugar µm Analytical KS x 00mm 7 µm Analytical KS , x 00mm 7 µm Analytical KS , x 00mm 7 7µm Analytical KS , x 00mm 7 7µm Analytical KS-80 00,000, x 00mm 76 7µm Analytical KS ,000, x 00mm 77 related product Doing carbohydrate analysis? ELSD offers greater sensitivity than RI. See pages. related product Looking for column heaters? See pages 6 and

58 Shodex ODP HP Columns High-Efficiency Polymer-Based Reversed-Phase Column Column efficiency is comparable with that of silica-based ODS columns Better retention of highly polar substances comparing to ODS columns Long column life even with high protein content samples: ODP HP prevent protein adsorption that causes ODS column degradation Excellent peak shape using low salt mobile phase, ideal for microbore High ph stability ODP HP series is a polymer-based [poly(hydroxymethacrylate)] column for reversed-phase chromatography. The efficiency of ODP HP is improved over most resin-based columns, with typical theoretical plate number >6,000 per meter. ODP HP Columns Packing Format i.d. x Length Part No. ODP HP-B, µm Analytical.6 x 0mm F7600 ODP HP-D, µm Analytical.6 x 0mm F7600 ODP HP-E, µm Analytical.6 x 0mm F7600 ODP HPG-A, µm Analytical.6 x 0mm F6700 ODP HP-B, µm Analytical.6 x 0mm F7600 ODP HP-D, µm Analytical.6 x 0mm F7600 ODP HPG-A, µm Analytical.6 x 0mm F670 ODP HP Columns Specifications Packing Theoretical Plate Number (per column) Theoretical Plate Number (per meter) Particle Size ph Range i.d. x Length ODP HP-B 00 70,000 µm.6 x 0mm ODP HP-D,000 86,000 µm.6 x 0mm ODP HP-E 7,000 68,000 µm.6 x 0mm ODP HPG-A guard column guard column µm.6 x 0mm ODP HP-B ,000 µm.0 x 0mm ODP HP-D ,600 µm.0 x 0mm ODP HPG-A guard column guard column µm.0 x 0mm hplc columns large molecule Drug in Biological Fluid Microbore is effective for the high sensitivity analysis of drugs; however, when protein is present and enters the MS (Mass detector), it contaminates the MS or suppresses ionization of the sample. Often pretreatment does not remove protein thoroughly. Drugs in biological fluid are hard to analyze because protein co-elutes with the component of interest. The target drug receives ion suppression from the protein and appears as a small peak. ODP HP can separate the target drug from protein by eluting protein early. The result of barbital analysis with BSA using microbore is shown as below. Barbital was introduced into the MS by a switching valve after BSA was eluted, and barbital was detected without any influence of ion suppression.,800k,600k,00k,00k,000k 800K 600K 00K 00K 0K 0K 0K 00K 90K 80K 70K 60K 0K 0K 0K 0K 0K 0 Valve Switch Barbital 0K 0K 0K 00K 90K 80K 70K 60K 0K 0K 0K 0K 0K 0 Valve Switch Barbital,800K BSA,600K,00K MS,00K,000K 800K 600K 00K 00K 0 UV 0 UV Min Min. MS more info For other Shodex columns, including KW00, KF-00, SB-800, KF-00, KF-600, KF-800, NHP, Ion Exchange, affinity, and sugar columns, visit. 8

59 Waters Spherisorb Columns All Waters columns are packed by Grace to our highest QC standards. Spherisorb columns have 80Å pores and 0m /g surface area. ODS- is unendcapped with a 6% carbon load, and ODS- is endcapped with a % carbon load. Spherisorb HPLC Columns Packing Format i.d. x Length Part No. Waters Fittings Part No. ODS-, µm Analytical.6 x 0mm 8 8 Analytical.6 x 0mm ODS-, µm Analytical.6 x 00mm Analytical.6 x 0mm ODS-, µm Analytical.6 x 0mm 8 87 Analytical.6 x 0mm C8, µm Analytical.6 x 00mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 0mm C6, µm Analytical.6 x 0mm 8 87 Analytical.6 x 0mm 8 8 Phenyl, µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm Analytical.6 x 0mm Amino, µm Analytical.6 x 0mm Analytical.6 x 0mm Silica, µm Analytical.6 x 00mm Analytical.6 x 0mm 8 86 Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm SAX, µm Analytical.6 x 0mm SCX, µm Analytical.6 x 0mm Spherisorb All-Guard Cartridges* Packing i.d. x Length Qty. Part No. ODS-, µm.6 x 7.mm 968 ODS-, µm.6 x 7.mm 969 C8, µm.6 x 7.mm 960 C6, µm.6 x 7.mm 96 Phenyl, µm.6 x 7.mm 966 Cyano, µm.6 x 7.mm 96 Amino, µm.6 x 7.mm 96 Silica, µm.6 x 7.mm 967 SAX, µm.6 x 7.mm 968 SCX, µm.6 x 7.mm 967 All-Guard Cartridge Holder ea 800 (Includes Direct-Connect Column Coupler) *All-Guard holder required. Other particle sizes available. more info If you like Spherisorb, try Grace s lower cost Allsphere on page 9. Beckman Ultrasphere Columns Low metal content for symmetrical peaks Narrow particle size range for high efficiency Maximum surface coverage for long column lifetimes All Ultrasphere columns are packed by Grace to our highest QC standards. Ultrasphere columns are highly endcapped to reduce silanol interactions and have a narrow particle size range for excellent resolving power. Tricyclic Antidepressants Min Doxepin. Desipramine. Nortriptyline. Imipramine. Amitriptyline 6. Trimipramine 7. Clomipramine Column: Ultrasphere C8, µm,.6 x 0mm (Part No. 0) Mobile Phase: 0mM Triethylamine, ph.:acetonitrile (7:9) Flow Rate:.0mL/min Detector: UV at 6nm Ultrasphere HPLC Columns Packing Format i.d. x Length Part No. C8, µm Analytical.6 x 7mm C8, µm Microbore.0 x 0mm 790 Microbore.0 x 0mm Analytical.6 x 0mm 0 Analytical.6 x 0mm 9 Prep 0 x 0mm 8 C8-IP, µm Analytical.6 x 0mm Analytical.6 x 0mm C8, µm Analytical.6 x 0mm Analytical.6 x 0mm Cyano, µm Analytical.6 x 0mm 070 Analytical.6 x 0mm 07 Silica, µm Analytical.6 x 0mm Analytical.6 x 0mm Ultrasphere Guard Columns Description i.d. x Length Qty. Part No. C8, µm.6 x mm ea 6 C8-IP, µm.6 x mm ea C8, µm.6 x mm ea Cyano, µm.6 x mm ea 8 Silica, µm.6 x mm ea 0 Ultrasphere guard columns connect directly to the analytical column with a zero dead volume connector, or a small piece of tubing with finger tight fittings. No holder required. 8

60 ZirChrom Columns Better ph and thermal stability than silica-based columns More efficient than polymer-based columns Widest range of reversed-phase selectivities ZirChrom Separations, Inc. produces HPLC columns based on µm zirconia (zirconium oxide) particles. These columns are more rugged and durable than silica-based columns and more efficient than polymer-based columns. ZirChrom normal-phase and reversed-phase columns can be used across the entire ph range ( ) and are thermally stable to 00 C. Additionally, zirconia s silanol-free surface produces sharp, symmetrical peaks for amines without mobile phase modifiers. ZirChrom Packings Packing Functional Group Mode DiamondBond -C8 Octadecylsilane Reversed-Phase PBD Polybutadiene Reversed-Phase & Secondary Ionic PS Polystyrene Reversed-Phase (Low Retention) CARB Graphitic Carbon Reversed-Phase (High Retention) PHASE Zirconia Normal-Phase EZ Reversed-Phase MS Reversed-Phase DiamondBond -C8 ideal for microbore applications and acidic compounds. ph range ZirChrom -PBD for general purpose applications and basic compounds; similar to ODS for non-electrolytes. ph range ZirChrom -PS ideal for highly aqueous mobile phases; an alternative to ODS selectivity. ph range ZirChrom -CARB for diastereomers/geometric isomers; greatest difference in selectivity compared to ODS. ph range ZirChrom -Phase the bare zirconia particle. The columns are used as a normal-phase alternative to silica. ph range ZirChrom -EZ unique selectivity with mobile phase selectivity. ph range 0 ZirChrom -MS low column bleed makes it ideal for microbore applications. ph range 0 ZirChrom Specifications Pore Size 00Å Particle Size µm, µm Surface Area 0m /g ph Range (EZ and MS 0) Max. Temperature 00 C EZ and MS HPLC Columns Unique selectivity on a zirconia platform Mobile phase flexibility, including microbore compatible buffers Stable from ph 0 The deactivation of Lewis acid sites on the zirconia surface allows for the chromatography of Lewis base analytes using volatile mobile phase additives, including conventional microbore compatible buffers such as acetate and formate. ZirChrom HPLC Columns Packing Format i.d. x Length Part No. DiamondBond, µm Microbore. x 00mm DB0-0 Microbore. x 0mm DB0- Analytical.6 x 0mm DB0-06 Analytical.6 x 00mm DB0-06 Analytical.6 x 0mm DB0-6 PBD, µm Microbore. x 00mm ZR0-0 Microbore. x 0mm ZR0- Analytical.6 x 0mm ZR0-06 Analytical.6 x 00mm ZR0-06 Analytical.6 x 0mm ZR0-6 PS, µm Microbore. x 00mm ZR09-0 Microbore. x 0mm ZR09- Analytical.6 x 0mm ZR09-06 Analytical.6 x 00mm ZR09-06 Analytical.6 x 0mm ZR09-6 CARB, µm Microbore. x 00mm ZR0-0 Microbore. x 0mm ZR0- Analytical.6 x 00mm ZR0-06 Analytical.6 x 0mm ZR0-6 PHASE, µm Microbore. x 00mm ZR0-0 Microbore. x 0mm ZR0- Analytical.6 x 00mm ZR0-06 Analytical.6 x 0mm ZR0-6 EZ, µm Microbore. x 0mm EZ0-0 Microbore. x 00mm EZ0-0 Microbore. x 0mm EZ0- Microbore. x 0mm EZ0- Analytical.6 x 0mm EZ0-06 Analytical.6 x 00mm EZ0-06 Analytical.6 x 0mm EZ0-6 Analytical.6 x 0mm EZ0-6 EZ, µm Microbore. x 0mm EZ0-0 Microbore. x 00mm EZ0-0 Microbore. x 0mm EZ0- Microbore. x 0mm EZ0- Analytical.6 x 0mm EZ0-06 Analytical.6 x 00mm EZ0-06 Analytical.6 x 0mm EZ0-6 Analytical.6 x 0mm EZ0-6 MS, µm Microbore. x 0mm MS0-0 Microbore. x 00mm MS0-0 Microbore. x 0mm MS0- Microbore. x 0mm MS0- Analytical.6 x 0mm MS0-06 Analytical.6 x 00mm MS0-06 Analytical.6 x 0mm MS0-6 Analytical.6 x 0mm MS0-6 MS, µm Microbore. x 0mm MS0-0 Microbore. x 00mm MS0-0 Microbore. x 0mm MS0- Microbore. x 0mm MS0- Analytical.6 x 0mm MS0-06 Analytical.6 x 00mm MS0-06 Analytical.6 x 0mm MS0-6 Analytical.6 x 0mm MS0-6 8

61 Vydac Large Molecule Columns Vydac has always been a trusted name in bioseparations, now, with technology acquired by Grace over the past few years, we have expanded this expertise further. New Grace large molecule columns range from nano, capillary to micro, analytical to preparative columns. hplc columns large molecule Separate biomolecules from small peptides to large intact proteins with the Vydac family of products which includes reversedphase, normal-phase, ion-exchange, and affinity phases. Our extensive applications library offers solid method development guidance, and our technical experts provide insight to even the most unusual separation challenges. Whether your primary analysis consideration is speed, MS compatibility, resolution, or recovery, we have a solution. Column Selection for Polypeptides The reversed-phase column for a polypeptide separation should be selected based on the hydrophobicity of the polypeptide being chromatographed and molecular weight as a secondary consideration. Molecular Weight 000 Increasing hydrophilicity 0,000 C 8 = GraceAlpha C 8 = 8TP, 8TP, 8MS C 8 = 08TP, 08MS C = TP, MS Increasing hydrophobicity 00,000 Diphenyl = 9TP Phase Simple Enzymatic Digests (< proteins) Complex Enzymatic Digests (> proteins) Biomolecules 0-K MW Biomolecules -0K MW Biomolecules >0K MW Un-denatured, Intact Proteins Antibodies Oligonucleotides Oligosaccharides Comments 8MS (C8) See page 8 87 Polymeric bonding highest hydrophic interaction and unique geometric selectivity 8MS (C8) See page 8 87 Monomeric bonding offers increased peptide interaction and generally yields higher peak counts. Different selectivity compared to 8MS. 08MS (C8 See page 8 87 Lower hydrophobicity is better for larger molecules MS (C) See page 8 87 Ideal for hydrophobic proteins, or when minimal organic is desired 9MS (Diphenyl) See page 8 87 Lowest capacity, highly selective for proteins with aromatic sidechains Everest C8 See page Maximum surface coverage for highest resolution of complex samples ProZap See page 9 9 Sub µm, in Expedite format optimized for fast analysis 8TP (C8) See page 90 9 First generation media with extensive applications library TP (C) See page 90 9 First generation media with extensive applications library related products Looking for large molecule prep columns? See pages

62 Vydac MS Introduction New Generation Columns with Unsurpassed Resolution, Sensitivity, and Recovery Unique selectivity reveals peaks otherwise masked by other C8 columns Excellent peak shape with little or no TFA High protein recoveries make scale-up easy Vydac MS columns are the latest development in the ongoing effort to provide the best reversedphase HPLC columns for biomolecule. A surface treatment and propriatary bonding process gives Vydac MS columns a unique selective not found anywhere else. A variety of reversed phases makes this product line suitable for small peptides to large intact, undenatured proteins. MS Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code 08MS C8 Silica Spheroidal µm 00Å 70m /g % Polymeric Yes L7 MS C Silica Spheroidal µm 00Å 70 0m /g % Polymeric Yes L6 8MS C8 Silica Spheroidal,, 0, 0 µm 00Å 60 0m /g 8% Polymeric Yes L 8MS C8 Silica Spheroidal µm 00Å 70m /g % Monomeric Yes L 9MS Di-Phe Silica Spheroidal µm 00Å 70m /g % Polymeric Yes Unique Selectivity A sample of bovine fetuin, a 6kD glycoprotein, was digested with trypsin. Some of the sample components interfere with the peptide separation on the Competitor and Competitor columns and appear as a chromatographic hump with peaks riding on top. The unique selectivity of Vydac MS columns solves these separation problems Min kd Protein. Non-myristoylated p. Myristoylated p. Triton X (Surfactant) Vydac MS C Column Competitor C Column Competitor C Column Competitor C Column Competitor C Column Columns: Vydac MS C MS, Competitor C, Competitor C, Competitor C, and Competitor C (all 00Å, µm, except µm for Competitor,.6 x 0mm) Flow Rate: Mobile Phase: Transmembrane Protein p Columns.0mL/min A: 0.% v/v TFA in HO B: 0.08% v/v TFA in Acetonitrile Gradient Vydac, Comp., and Comp. : Time: 0 0 %B: Gradient Comp. and Comp. : Time: %B: Detector: UV at nm Column Temp: Ambient, C Injection Vol: 0µL Unsurpassed Resolution and Peak Symmetry Vydac MS C columns provide the overall best performance for hgh and desamido hgh analysis. Competitive columns show significant, undesirable interaction of hgh with the stationary phase. Human Growth Hormone (hgh) Assay Columns. Desamido HGH. HGH Vydac MS C Column Competitor C Column Competitor C Column Competitor C Column Min. Columns: All 00Å, µm,.6 x 0mm Mobile Phase: Isocratic, 7% 0mM Tris, ph 7., 9% n-propanol Flow Rate: 0.mL/min Detector: UV at 0nm Column Temp: C Injection Vol: 0µL of a mg/ml preparation more info For additional protein and peptide applications, see application section pages 9. more applications To view our complete searchable chromatogram database visit /chromdb/ 700 hplc columns large molecule Reptilian reovirus RRV p sample courtesy of Drs. Roberto J. de Antueno and Roy Duncan, Dalhousie University, Halifax, Nova Scotia. 8

63 hplc columns large molecule Vydac MS Columns Excellent Peak Shape with Little or No TFA It is common practice for protein and peptide separations to include an acidic modifier such as TFA in the mobile phase. TFA masks basic entities, reducing mixed-mode retention, and improving peak symmetry. TFA also changes retention and selectivity for different analytes, and its concentration can be adjusted to optimize a separation. Unfortunately TFA is UV absorbent and contributes background at low UV wavelengths. Also It is especially problematic with electrospray MS where it interferes with ion generation, called quenching, and reduces MS sensitivity. 0.6 AU Columns: 0.% TFA Flow Rate: Mobile Phase: Gradient: Detector: Peptides on 8MS C8 Reversed-Phase 0.0% TFA 0.0% TFA 0.0% TFA. Neurotensin -8 (fragment). Oxytocin. Angiotensin II. Neurotensin Min. Vydac C8, µm,.6 x 0mm (Part No. 8MS).mL/min A: % Acetonitrile in Water with TFA as indicated (v/v) B: 9% Acetonitrile in Water with TFA as in A Linear from 0 to 0% B over 0min, then to 00% B in min UV at 0nm Hydrophobic Proteins Transmembrane proteins are hydrophobic proteins which bind to cell membranes and are particularly difficult to separate. Vydac MS columns provide excellent selectivity and peak shape for these molecules. In this case, a hydrophobic transmembrane protein was separated from a synthetic myristoylated derivative and other cellular components. Tryptic Digest of Fetuin on 00Å C8 Columns Vydac MS C8 Column Competitor C8 Column Competitor C8 Column Min. Columns: Vydac MS C8 8MS, Competitor C8, and Competitor C8 (all 00Å, µm,.6 x 0mm) Flow Rate:.0mL/min Mobile Phase: A: 0.% v/v TFA in HO B: 0.08% v/v TFA acid in Acetonitrile Gradient: Time: %B: Detector: UV at nm Column Temp: Ambient, C Injection Vol: µl of a 6 µg/µl digest (80 µg total peptide load) The Vydac column provides better separation and recovery for a highly hydrophobic membrane protein (RRV p) and its fatty acid modified (myristolyated) form, a component of a potentially new vaccine delivery system. High Protein Recovery for Higher Sensitive, Ideal for Preparative Chromatography Surface chemistry reduces adsorption of proteins for higher recoveries and also increases mass loading. more applications To view our complete searchable chromatogram database visit /chromdb/ % Recovery at Low Protein Load: Vydac MS C vs. Competitors Loading Amount: µg of Cytochrome C % Recovery Vydac MS Competitor Competitor 0 Vydac MS C column provides more than 0% more loading of Cytochrome C. 86

64 Vydac MS Columns Vydac MS Analytical Columns Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge MS C µm.0mm MS0 MS0 MS MS GKMS GDMS.mm MS0 MS0 MS MS GKMS GDMS.mm MS GKMS GDMS.6mm MS0 MS0 MS MS GKMS GDMS 08 MS C8 µm.0mm 08MS0 08MS0 08MS 08MS 08GKMS 08GDMS.mm 08MS0 08MS0 08MS 08MS 08GKMS 08GDMS.mm 08MS 08GKMS 08GDMS.6 mm 08MS0 08MS0 08MS 08MS 08GKMS 08GDMS 8 MS Polymeric C8 µm.0mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS.mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS.mm 8MS 8GKMS 8GDMS.6mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS 8 MS Monomeric C8 µm.0mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS.mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS.mm 8MS 8GKMS 8GDMS.6mm 8MS0 8MS0 8MS 8MS 8GKMS 8GDMS 9 MS Diphenyl µm.0mm 9MS0 9MS0 9MS 9MS 9GKMS 9GDMS.mm 9MS0 9MS0 9MS 9MS 9GKMS 9GDMS.mm 9MS 9GKMS 9GDMS.6mm 9MS0 9MS0 9MS 9MS 9GKMS 9GDMS A guard kit includes a holder and one guard cartridge; Guard cartirdge units include two guard cartridges. hplc columns large molecule Vydac MS Nano and Capillary Columns i.d. 0mm 00mm 0mm 0mm MS C µm 7µm MS.070 MS.070 MS.07 MS.07 0µm MS.0 MS.0 MS. MS. 00µm MS.0 MS.0 MS. MS. 00µm MS.0 MS.0 MS. MS. 08 MS C8 µm 7µm 08MS MS MS.07 08MS.07 0µm 08MS.0 08MS.0 08MS. 08MS. 00µm 08MS.0 08MS.0 08MS. 08MS. 00µm 08MS.0 08MS.0 08MS. 08MS. 8 MS Polymeric C8 µm 7µm 8MS.070 8MS.070 8MS.07 8MS.07 0µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. µm 7µm 8MS.070 8MS.070 8MS.07 8MS.07 0µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. 8 MS Monomeric C8 µm 7µm 8MS.070 8MS.070 8MS.07 8MS.07 0µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. 00µm 8MS.0 8MS.0 8MS. 8MS. 9 MS Diphenyl µm 7µm 9MS.070 9MS.070 9MS.07 9MS.07 0µm 9MS.0 9MS.0 9MS. 9MS. 00µm 9MS.0 9MS.0 9MS. 9MS. 00µm 9MS.0 9MS.0 9MS. 9MS. Vydac MS Guard Cartridges Packing i.d. x Length Qty. Part No. C8 Polymeric, µm* Capillary Guard** 0.0 x 0mm ea 8MSC x 0mm ea 8MSC00.0 x 0mm 8GDMS C, µm* Capillary Guard** 0.0 x 0mm ea MSC x 0mm ea MSC00.0 x 0mm GDMS *All-Guard holder required. Other particle sizes available. **.µm and µm particles and other dimensions are available. Vydac MS Guard Cartridges (continued) Packing Qty. Part No. Capillary Guard Cartridge Holder Guard Holder for 0.00mm and 0.0mm Guards ea GR-70E Guard Holder for 0.00mm and 0.00mm Guards ea GR-70A Guard Holder for mm Guards ea GCH 87

65 hplc columns large molecule Vydac Everest Columns Introduction High Resolution for Complex Peptide Samples High resolution of complex peptide digests Unique selectivity for both hydrophilic and hydrophobic peptides Exceptional microbore sensitivity with little or no TFA in the mobile phase Everest columns provide exceptionally high resolution for complex peptide digest separations. Unique selectivity and sensitivity are the result of new bonding technology that improves C8 surface coverage and deactivates residual silanols. Previously, the best 00Å C8 chemistries have had carbon coverage in the.8 to.6µmol/m range. Everest C8 coverage is in excess of µmol/m and approximates the theoretical limit based on surface area. This increased shielding of the base silica increases column life and reduces the amount of TFA required to shield the silica. Everest Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code 8EV C8 Silica Spheroidal, 0, 0, 0µm 00Å 70 0m /g 6% Monomeric Yes L Tryptic Digest of Bovine Serum Albumin Everest Column 0 peaks Commercial Column C 9 peaks Commercial Column D 96 peaks Intensity (x 0 6 cps) Base Peak Chromatogram of Tryptic Digest of BSA Min. Column: Vydac Everest C8, 00Å, µm,.6 x 0mm Commercial 00å c8 columns.6 x 0mm (Part No. 8EV) Mobile Phase: A: 0.% TFA in Water B: 0.08% TFA in Acetonitrile Gradient: Time: %B: Flow Rate:.0mL/min Detector: UV at nm Everest columns outperform competitor columns at higher peptide loads, by providing higher-resolution separations (average of 7% higher peak counts than competitor columns tested) Min. Column: Vydac Everest C8, 00Å, µm, 00µm x 0mm; (Part No. 8EV.) Mobile phase: A: 0.% v/v Formic Acid in Water B: 0.% v/v Formic Acid in Acetonitrile Gradient: Time: %B: Flow Rate: µl/min Column Temp: C Detector: MS + ESI The Everest column performs exceptionally well with no TFA in the mobile phase, ensuring excellent microbore sensitivity. Everest 00Å C8 Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge Everest Analytical Columns 8EV C8 µm.0mm 8EV0 8EV0 8EV 8EV 8GKEV 8GDEV.mm 8EV0 8EV0 8EV 8EV 8GKEV 8GDEV.6mm 8EV0 8EV0 8EV 8EV 8GKEV 8GDEV Everest Capillary Columns 8ZU C8 µm 7µm 8EV.070 8EV.070 8EV.07 8EV.07 0µm 8EV.0 8EV.0 8EV. 8EV. 00µm 8EV.0 8EV.0 8EV. 8EV. 00µm 8EV.0 8EV.0 8EV. 8EV. A guard kit includes a holder and one guard cartridge. Guard cartirdge units include two guard cartridges. Titamium frits are standard in column diameters.6mm and smaller. Everest Guard Cartridges Packing i.d. x Length Qty. Part No. C8, µm* Capillary Guard** 0.0 x 0mm ea 8EVC x 0mm ea 8EVC00.0 x 0mm 8GDEV *All-Guard holder required. Other particle sizes available. **.µm and µm particles and other dimensions are available. Everest Guard Cartridges (continued) Packing Qty. Part No. Capillary Guard Cartridge Holder Guard Holder for 0.00mm and 0.0mm Guards ea GR-70E Guard Holder for 0.00mm and 0.00mm Guards ea GR-70A Guard Holder for mm Guards ea GCH 88

66 Vydac Everest Columns Tryptic Digest of Cytochrome C Columns: Flow Rate: Mobile Phase: Everest 8EV and competitor 00Å reversed-phase columns, all C8, µm,.6 x 0mm.0 ml/min A: 0.% v/v TFA in water B: 0.08% v/v TFA in ACN Gradient: Time: %B: hold 90% B for 0 min Detector: UV at nm Everest Column Competitor Column B Competitor Column D Competitor Column E Competitor Column F Number of Well-Resolved Peptide in Highlighted Areas 7 6 hplc columns large molecule Min. For a tryptic digest of Cytochrome C, an Everest column offers unique selectivity which allows the best separation of a group of hydrophilic and hydrophobic peptides. To assess resolution, chromatograms of tryptic digests of several proteins on an Everest column and several other commercial 00Å C8 reversed-phase columns were compared. Peak counts were based on detection by automated chromatography software with parameters set identically for all columns. In the analyses of a tryptic digest of cytochrome c, specific regions of the chromatogram were examined for the number of peaks resolved. The Everest column demonstrated unique selectivity for both hydrophilic and hydrophobic peptides. Tryptic Digest of Fetuin (a Glycoprotein) Everest Column 98 peaks Competitor Column A 97 peaks Columns: Everest 8EV and competitor 00Å reversed-phase columns, all C8, µm,.6 x 0mm Flow Rate:.0 ml/min Mobile Phase: A: 0.% v/v TFA in water B: 0.08% v/v TFA in ACN Gradient: Time: %B: Detector: UV at nm Competitor Column B 90 peaks Competitor Column C 9 peaks Competitor Column D 89 peaks Min. A tryptic digest of fetuin injected at high load on an Everest column provided higher peak counts compared to four commercial columns. Peak numbers shown are the average of three separations on each column. more applications To view our complete searchable chromatogram database visit /chromdb/ 89

67 hplc columns large molecule Vydac TP Columns Industry Standard for Polypeptide Separations Referred in over 9000 patents, Vydac 00Å TP is the industry-standard, for peptide, protein, and large molecule separations Polymeric bonded phases have exceptionally long column lifetime and negligible phase leaching Extensive applications library based on over two decade s experience Vydac TP reversed-phase material consists of aliphatic groups bonded to the surface of 00Å pore diameter silica. The large pores of the 00Å TP silica give polypeptide molecules complete access to the interior of the silica pores. The unique process by which we manufacture Vydac TP silica results in high-purity, synthetic silica with carefully controlled characteristics. Vydac TP silica is the standard that has defined large pore HPLC silica for polypeptide separations for nearly two decades. Vydac TP Columns Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code 0TP Sil Silica Spheroidal, 0, 0, 0µm 00Å 70 0m /g unbonded L 0TP C8 Silica Spheroidal, 7, 0, 0, 0µm 00Å 70 90m /g 8% Polymeric No L 0TP C8 Silica Spheroidal,, 0µm 00Å 60 90m /g 9% Polymeric No L 08TP C8 Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g % Polymeric Yes L7 TP C Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g % Polymeric Yes L6 8TP C8 Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g 8% Polymeric Yes L 9TP Di-Phe Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g % Polymeric Yes 8TP C8 Silica Spheroidal,, 7, 0, 0, 0µm 00Å 60 0m /g % Monomeric Yes L Vydac 8TP C8 Columns Vydac 8TP is a polymerically bonded endcapped n-octadecyl reversed-phase based on 00Å TP silica. Applications Vydac 8TP reversed-phase columns are recommended for the separation of: Small polypeptides less than MW Enzymatic digest fragments Natural and synthetic peptides Multi-ring compounds Specific examples include: Tryptic digests S. aureus V8 digests Synthetic peptides Natural peptides Peptide studies Standard Human Apolipoproteins Min. Column: Vydac 8TP, C8, µm,.6 x 0mm Flow Rate:.mL/min Column Temp: 0 C Mobile Phase: A: % ACN/0.% TFA in Water B: 8% ACN/0.% TFA in Water Gradient: 0 00% B in min apoc-iiia. apoc-iiib. apoc-iia. apoc-iiic. apoc-i 6. apoc-iib 7. apoa-ia 8. apoa-ib 9. apoa-iic 0. apoa-iia. apoa-iib The standard was human insulin Vydac 08TP C8 Reversed-Phase Vydac 08TP is a polymerically bonded endcapped n-octyl reversed-phase based on 00Å TP silica. Separation of a Tryptic Digest on 08TP Reversed-Phase Column Applications Vydac 08TP reversed-phase columns are recommended for the separation of: Polypeptides up to 0,000 0,000 MW Enzymatic digest fragments Natural and synthetic peptides Min. Column: Flow Rate: Gradient: Vydac 08TP, C8, µm,.6 x 0mm.0mL/min 0 0% ACN with 0.% aqueous TFA over 60min 90

68 Vydac TP C Vydac TP is a polymerically bonded endcapped n-butyl reversed-phase based on 00Å TP silica. ATP is a less extensively endcapped C that has been found more suitable for resolution of degradation products in analysis of biosynthetic human growth hormone. Applications Glycoproteins Hemoglobin variants Histones Human growth hormone Insulin variants Membrane proteins Vydac 9TP Diphenyl Reversed-Phase Vydac 9TP is a polymerically bonded endcapped diphenyl reversed-phase based on 00Å TP silica. It combines moderate retentivity with unique selectivity. Applications Polypeptides with aromatic side chains Large, hydrophobic proteins Membrane-spanning peptides Lipid peptides Fusion proteins from inclusion bodies Insulin Variants Min. Column: Vydac C, µm,.6 x 0mm (Part No. TP) Gradient: 7 0% ACN with 0.% TFA over min From J. Rivier and R. McClintock, J. Chrom. 68, -9 (98) Chicken. Bovine. Ovine. Rabbit. Human 6. Porcine 7. Rat I 8. Rat II Comparison of Polypeptide Separations on C8, C, and Diphenyl Reversed-Phase Columns 8TP (C8) TP (C) 9TP (diphenyl). H. TH6 hplc columns large molecule Min. Columns: Gradient: Sample: Vydac 8TP (C8), TP (C) and 9TP (diphenyl), all µm,.6 x 0mm 0 90% ACN with 0.% TFA over 0min 8-residue helical peptide () and six-helix template assembled synthetic protein () Vydac 8TP C8 Reversed-Phase Vydac 8TP is a monomerically bonded endcapped n-octadecyl reversed-phase based on 00Å TP silica. Applications Monomerically bonded C8 provides an alternative to 8TP polymeric C8 with subtle differences in selectivity. The combination of these adsorbents can reveal analytes that may be hidden on a single C8 column. more info For more protein and peptide applications, see the application section pages 9. more applications To view our complete searchable chromatogram database visit /chromdb/ 8TP polymeric C8 6 8TP monomeric C8, 6 Columns: Flow Rate: Gradient: Detector: Synthetic Peptides, 8, Min.. GY. VYV. Neurotensin fragment -8 (pelyenkpr). Ac-RGGGGLGLGK-NH. RGAGGLGLGK-NH 6. Ac-RGAGGLGLGK-NH 7. Ac-RGVGGLGLGK-NH 8. Oxytocin (CYIQNCPLG-NH) 9. Met Enkephalin (YGGFM) 0. Bradykinin (RPPGFSPFR). Ac-RGVVGLGLGK-NH. Neurotensin Fragment 8- (RRPYIL). Angiotensin II (DRVYIHPF). Leu Enkephalin (YGGFL). Neurotensin (pelyenkprrpyil) 6. Angiotensin I (DRVYIHPFHL) Vydac 8TP and 8TP, both C8, µm,.6 x 0mm.0mL/min 0 0% ACN with 0.% TFA (w/v) over 0min UV at 0nm 9

69 hplc columns large molecule Vydac TP Columns Vydac TP Analytical Columns Columns Recommended Guards Particle Size i.d. 0mm 00mm 0mm 0mm Guard Kit Guard Cartridge 8TP C8 µm.6mm 8TP0 8TP0 8GK 8GD µm.0mm 8TP0 8TP0 8TP 8TP 8GK 8GD.mm 8TP0 8TP0 8TP 8TP 8GK 8GD.mm 8TP0 8TP0 8TP 8TP 8GK 8GD.6mm 8TP0 8TP0 8TP 8TP 8GK 8GD 08TP C8 µm.6mm 08TP0 08TP0 08GK 08GD µm.0mm 08TP0 08TP0 08TP 08TP 08GK 08GD.mm 08TP0 08TP0 08TP 08TP 08GK 08GD.mm 08TP0 08TP0 08TP 08TP 08GK 08GD.6mm 08TP0 08TP0 08TP 08TP 08GK 08GD TP C µm.6mm TP0 TP0 GK GD µm.0mm TP0 TP0 TP TP GK GD.mm TP0 TP0 TP TP GK GD.mm TP0 TP0 TP TP GK GD.6mm TP0 TP0 TP TP GK GD ATP C Columns µm.mm ATP.6mm ATP 9TP Diphenyl µm.6mm 9TP0 9TP0 9GK 9GD µm.0mm 9TP0 9TP0 9TP 9TP 9GK 9GD.mm 9TP0 9TP0 9TP 9TP 9GK 9GD.mm 9TP0 9TP0 9TP 9TP 9GK 9GD.6mm 9TP0 9TP0 9TP 9TP 9GK 9GD 8TP C8 µm.6mm 8TP0 8TP0 8GK 8GD µm.0mm 8TP0 8TP0 8TP 8TP 8GK 8GD.mm 8TP0 8TP0 8TP 8TP 8GK 8GD.mm 8TP0 8TP0 8TP 8TP 8GK 8GD.6mm 8TP0 8TP0 8TP 8TP 8GK 8GD NOTE: Additional column diameters and lengths are available on request. Please contact Grace Davison Discovery Sciences to discuss your requirements. A guard kit includes a holder and one guard cartridge; Guard cartirdge units include two guard cartridges. related products For prep Vydac TP Columns, see our prep section pages related products Looking for HPLC column prefilters? See page. related products Need high-pressure polymeric fittings? See pages for our full selection of high-pressure fittings. 667 technical assistance Contact Tech Support: Phone: (North America) contact.alltech@grace.com Online: related products Need HPLC tubing? See pages

70 Vydac ProZap C8 Ultra Fast Protein and Peptide Separations 0 times faster bioseparations than traditional column formats Ultra-high efficiency.μm packing Fast protein and peptide separations with conventional HPLC systems ProZap.μm, 00Å packings are ideal for fast bioseparations in life science applications. The combination of ProZap packings and short, 0mm Expedite column hardware delivers not only faster sample throughput, but also low back pressures suitable for conventional LC systems. Optimization of Gradient Parameters for Fast Protein Analysis Short ProZap columns are the perfect tool for fast reversed-phase protein separations. Under gradient conditions, longer columns only increase run time and do not increase resolution to improve the separation. Proteins adsorb at the head of the column and then desorb once the critical mobile phase concentration is reached. Since the proteins do not interact with the full length of the packed bed, short columns are sufficient for full resolution. Therefore, proteins of broad molecular weight ranges can be separated in less than one minute by combining short columns, higher flow rates, and fast, modified gradients. For best results high-pressure mixing should be used with fast gradients. ProZap C8 Specifications Phase Base Material Particle Shape Particle Size Pore Size Surface Area Carbon Load Phase Type Endcapped? USP L-code C8 ProZap Silica Spherical.µm 00Å 9m /g % Monomeric Yes L 779 hplc columns large molecule Proteins 6 7. Ribonuclease (7ng). Bovine Insulin (7ng). Cytochrome C (7ng). Lysozyme (ng). Bovine Serum Albumin (7ng) 6. Carbonic Anhydrase (7ng) 7. Ovalbumin (7ng) Min. Flow Rate: 0.8mL/min Backpressure: 8bar = 680psig Gradient: Time: %B: Min. Flow Rate: 0.mL/min Backpressure: 9bar = 80psig Gradient: Time: %B: Min. Flow Rate: 0.mL/min Backpressure: bar = 60psig Gradient: Time: %B: 7 7 Column: ProZap C8,. x 0mm, C8,.µm, 00Å HPLC System: High-pressure gradient, binary pump Mobile Phase: A: 0.% TFA in Water B: 0.08% TFA in Acetonitrile Detector: UV at 80nm Injection Vol: µl Injection Amt: 7ng each protein, except ng for lysozyme Column Temp: C Seven proteins were tested on a 0mm ProZap column. By increasing flow rate and reducing gradient time, total run time is reduced from. minutes to. minutes. more info For more information about the Expedite hardware format, see page. more applications To view our complete searchable chromatogram database visit /chromdb/ 6 9