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,, 7, Monomeric Yes L 0m /g, 7% 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 Silica Irregular, 7µm 00Å 00m /g Monomeric No L8 to silica) Diol Silica Spherical, 0µm 60, 0, 00, 00Å 80, 00, 00, Monomeric No L0 0m /g 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, No 0m /g 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, 00Å 60 0m /g % Polymeric Yes 0µm 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 Min. 00 mau 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 Min. 6. Uracil. Pyridine. Phenol. Methyl Alanine. -Butyl Benzoic Acid 6. Toluene Initial 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. 7 k' VisionHT Reversed-Phase VisionHT Selectivity 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 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 Min. No loss in efficiency. VisionHT Integral Guard System N=7K/m with guard without guard 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 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. 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 Min. The Alltima HP phases demonstrate excellent peak symmetry of highly basic analytes at ph 7.0. 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 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 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 k' 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 CN k' Toluene k' Butyl Benzoic Acid k' Amitriptyline Alltima HP Selectivity C8 EPS C8 C8 Amide C8 C8 AQ C8 HiLoad Alltima HP Packing 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 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 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. 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 ( 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 ( 9908) Alltima HP HILIC Column, μm,.6 x 0mm, ( 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 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. 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 Alltima HP CN ph.0 ph 7. 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 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 ( 87769) 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 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. 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/