HPLC Flash TLC GC SPE

Transcription

1 Chromatography Essentials HPLC Flash TLC GC SPE

2 Table of Contents HPLC Columns 5 HPLC Accessories 6 30 Flash Products 3 34 TLC Products GC Columns GC Accessories 46 5 SPE and Filtration Equipment Syringes Tubing 7 75 Vials 76 79

3 hplc columns & accessories Grace HPLC Column Solutions Address Today s Separation Challenges hplc columns This catalog features Grace s leading HPLC column technologies. You will find innovative chemistries, unsurpassed reliability, and hardware configurations that optimize your method requirements for speed, sensitivity, resolution or sample loading. Discover how our select range of small molecule columns, biomolecule-based offerings and ion chromatography technologies address your toughest separation challenges. Grace Featured Column Families: VisionHT Unique chemistries in sub to 0µm particles maximize performance and speed for UHPLC and HPLC. Everest High capacity for complex peptide mapping and proteomics applications. Alltima HP Premium quality, exceptionally stable columns, without phase bleed. ProZap High-speed protein and peptide separations. Prevail Compatible with 00% aqueous to 00% organic mobile phases. Vydac TP Popular, industry-standard bioseparations columns. GraceSmart High quality HPLC columns at an exceptional value. Alltech IC Wide variety for the separation of organic and inorganic anions and cations. Vydac MS Unique selectivity and recovery for protein isolation and characterization. Grace Expertise and Product Ownership Grace has been manufacturing silica and bonding it with unique chemistries for more than three decades. Our combined media production, column packing, and subμm particle expertise uniquely positions us to solve the challenges of modern chromatography laboratories. Partner with Grace and be assured our consistent, reliable chromatography products adhere to strict quality standards. Additional Column Brands Available While this catalog focuses on Grace leading HPLC columns, other popular brands are still available. Visit our website for a comprehensive listing. Alltima Adsorbosphere Genesis Grom Sil Certified in ISO 900, 4000 and 3485 procedures FDA CFR 80 for medical devices Full traceability from base silica to final product Extensive process control to ensure highest reproducibility Experienced global technical support and sales teams Allsphere Apex Apollo Brava Denali Econosphere Grom Sapphire Macrosphere Platinum ProSphere Vydac SP

4 hplc columns & accessories HPLC Column Hardware Formats HPLC Column Hardware Formats i.d. Format (mm) UHPLC Capillary Length (mm) System Compatibility Advantage Brand Availability VisionHT UHPLC system, LC/MS Capillary systems, LC/MS Ultra-low dead volume hardware, pressure rated to 8,000psig, allows 7X faster linear velocities. Highly specialized columns packed in small diameter fused silica tubing. Packed with subµm media, this hardware delivers the fastest separations with maximum efficiency. Extremely low sample dispersion offers best sensitivity. High-sensitivity and samplelimited applications. Alltima HP Vydac MS Everest hplc columns Expedite Low volume, highthroughput (HTP) systems Extremely short, low-volume columns that prevent sample diffusion. High-throughput and rapid resolution LC/MS applications packed with highly efficient.5µm media. VisionHT Alltima HP Prevail ProZap Microbore Standard HPLC optimized for low volume.mm diameter analytical columns requiring low volume systems. Low solvent consumption and improved sensitivity compared to traditional 4.6mm columns. VisionHT Alltima HP Prevail GraceSmart Vydac MS Everest IC Solvent Reducer Standard HPLC 3mm diameter analytical columns suitable for use on conventional HPLC. 50% solvent savings compared to traditional 4.6mm columns. No need to optimize system for low volume. VisionHT Alltima HP Prevail Vydac MS Vydac TP Analytical Standard HPLC Conventional 4.6mm i.d. with industry standard port configurations. Most common, traditional hardware format. VisionHT Alltima HP Prevail GraceSmart Vydac MS Everest Vydac TP IC Rocket Standard HPLC Large 7mm i.d. balances column volume with conventional LC system volume, and allows faster mobile phase flow rates. Ultra-fast separations on conventional HPLC systems. Unique hardware geometry reduces peak broadening and delivers excellent peak shapes. 5mL/min max flow rate reduces analysis time up to 80% compared to traditional analytical columns. VisionHT Alltima HP Prevail Preparative Preparative LC Large i.d. increases loading. Packed under high pressure for high efficiency LC purifications. VisionHT Vydac MS Vydac TP Capillary Guard UHPLC Guard 0 Guard system with reusable holder and disposable cartridges. Protects capillary columns and prolongs life. 5 Integral and stand-alone options. Protects UHPLC column investment, and prolongs life. Alltima HP Vydac MS Everest VisionHT All-Guard Guard system with reusable holder and disposable cartridges. Protects analytical and microbore columns and prolongs life. VisionHT Alltima HP Prevail Vydac MS Everest Vydac TP 3

5 hplc columns & accessories HPLC Phase Specifications and Usage hplc columns Columns for Small Molecules Brand VisionHT Phase C8 HighLoad Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties Application/Benefit 0 0 Yes Ultra high-purity silica, fully bonded. C8 Basic Proprietary Ultra-high purity silica. Controlled silica surface exposure gives dual mode separation with polar and nonpolar analytes. C8 Classic Yes Lower carbon load. Slight silica exposure. C8 Polar No High silica exposure, low carbon load. Uniform coverage of inert vicinal silanols. HILIC 0 0 No Polar phase with shorter equilibration times. Shipped in ACN/Water. Silica 0 0 No Traditional normal phase for use in 00% organic mobile phases. General purpose for broad range compounds, classic selectivity, high-capacity for hydrophobic compounds. Alternate reversed-phase selectivity. High polar retention especially with compounds having two or more polar groups. Excellent sensitivity and peak shape for basic compounds, without the need for acidified mobile phases. Reversed-phase separations with reduced bonding optimized for speed. Some additional polar retention. Unique polar selectivity. Low carbon load gives fastest reversed-phase elution times while retaining 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. Alltima HP C Yes Classic reversed-phase retention and selectivity. Routine applications. C8 EPS Yes Greater retention and enhanced Reversed-phase applications where C8 is too peak symmetry for polar retentive. compounds. Alternate selectivity to traditional reversed-phases. C8 Hi-Load Yes Highest carbon load for superior High resolution for complex samples. retention and loadability. C8 AQ Yes 00% water wettable. Applications requiring high aqueous mobile phases. C8 Amide Yes Polar-embedded phase with extremely low bleed. Compatible with microbore. C Yes Lower capacity compared to C8 phases. Cyano Yes Extremely stable, long life and reproducible. Basic compounds in neutral to alkaline ph, MS applications. Reversed-phase applications where C8 is too retentive. Ideal for basic drug analysis. Silica No Highly polar phase. General purpose normal phase applications. L3 HILIC 0 30 No Hydrophilic interaction Very polar analytes that cannot be retained by L3 chromatography uses small amounts of water for increased sensitivity with microbore applications. reversed-phase. Prevail C8 Select Yes Stable in 00% aqueous to 00% organic mobile phases. Same benefits as Prevail C8, but less retentive for polar compounds. L C Yes Stable in 00% aqueous to 00% organic mobile phases. Flexibility to switch between varied mobile phase conditions to suit a variety of applications. Excellent sensitivity for microbore applications. C Yes Stable C8 phase. Use for highly hydrophobic compounds that L7 retain too strongly on C8. Phenyl Yes Lowest hydrophobic capacity. Selective for aromatic compounds in a variety of L mobile phase conditions. Cyano Yes General purpose cyano suitable Rugged normal phase applications. L0 for normal or reversed-phase use. Amino No Stable in 00% aqueous to Use for carbohydrates or as a weak anion L8 00% organic mobile phases. exchanger. Silica No Highly polar phase. General purpose normal phase applications. L3 Organic Acid Yes Highly efficient silica-based, Separate common organic acids with acid-stable. unsurpassed resolution, speed and sensitivity. Lower cost than polymeric columns. Carbohydrate No Extremely stable hybrid phase. Excellent for mono- and oligosaccharides and ES sugar alcohols. GraceSmart C Yes High purity silica with monomeric C8 bonding offers classical reversed-phase selectivity. Excellent value for general purpose, routine applications. USP L-code L L L L L3 L3 L L L L L L7 L0 L L 4

6 hplc columns & accessories Columns for Large Molecules / Life Science Pore Surface Carbon Brand Phase Size (Å) Area (m /g) Load (%) Endcapped Chromatographic Properties Application/Benefit Vydac MS 8MS C Yes Polymeric bonding, highest hydrophobic Use for simple enzymatic digests (< proteins) interaction and unique geometric selectivity. or biomolecules 0 5K MW. 38MS C Yes Monomeric bonding offers increased peptide interaction and generally yields higher peak counts. 08MS C Yes Lower hydrophobicity is better for larger biomolecules. 4MS C Yes Lower capacity than C8 or C8, suitable for hydrophobic proteins or when minimal organic is desired. 9MS Yes Lowest capacity, aromatic functional Diphenyl groups. Everest 38EV C Yes Maximum surface coverage for highest resolution of complex samples. Use for same applications as 8MS, but offers different C8 selectivity. Ideal for biomolecules 5 0K MW. Ideal for biomolecules >0K MW, undenatured intact proteins, antibodies, oligonucleotides, human growth hormone. Highly selective for proteins with aromatic side chains. Complex enzymatic digests (> proteins). ProZap C Yes Subµm, 500Å wide pore. Ideal for fast, intact protein or peptide analysis. L Vydac TP 8TP C Yes First generation polymeric C8 media with Small polypeptides 4 5K MW, enzymatic digest L unique selectivity. fragments, natural and synthetic peptides, multi-ring compounds. 38TP C Yes First generation monomeric C8 media. Use for same applications at 8TP, but offers L different C8 selectivity. 08TP C Yes Less hydrophobic than C8TP media. Polypeptides 0 0K MW. L7 4TP C Yes First generation C4 media. Glycoproteins, hemoglobin variants, histones, insulin variants, membrane proteins. L6 9TP Diphenyl Yes Lowest capacity first generation diphenyl media. Reversed-Phase Column Comparisons Polypeptides with aromatic side chains, large hydrophobic proteins, membrane-spanning peptides, lipid peptides, fusion proteins from inclusion bodies. USP L-code L Based on the widely accepted work and test procedures of Drs Lloyd Snyder and John Dolan,, 3, Grace developed this tool to show relative selectivity comparisons for commercially available columns. This graph plots the indicators for hydrophobicity versus the cation exchange capacity at ph 7. Hydrophobicity is often the primary analyte interaction and indicates overall capacity. The cation exchange capacity measures secondary interactions that take place and can greatly influence the retention of polar analytes. L L7 L6 L L L hplc columns VisionHT C8 Polar VisionHT C8 Basic Acquity HSS C8 SB Extended Polar Retention Prevail Amide Prevail C8 Universally Retentive Increasing Polar Retention Alltima HP C8 Amide Alltima HP C8 EPS Synergi Polar-RP Zorbax SB-AQ To find columns that will produce alternate separation profiles, choose points that are in opposite quadrants. Prevail Select C8 GraceSmart C8 Atlantis T3 VisionHT C8 Classic Hypersil GOLD aq Hypersil GOLD Acquity BEH Shield RP-8 Kinetex XB-C8 Gemini C8 Acquity HSS T3 Kinetex C8 Ascentis ACE 5 C8 RP-Amide XTerra MS C8 Luna C8 Zorbax SB C8 To find columns with similar selectivity, choose points that are close together. Inertsil ODS- Zorbax Rx-8 Hypersil ODS- Hypersil BDS C8 Pinnacle DB C8 Alltima HP C8 VisionHT C8 HighLoad XBridge C8 Symmetry C8 Zorbax Eclipse XDB-C8 Acquity HSS C8 Zorbax Extend C8 Kromasil 00 5 C8 XTerra C8 RP Alltima HP C8 High Load Low Capacity Increasing Hydrophobic Retention Traditional RP Selectivity Data courtesy of Lloyd Snyder and John Dolan. The Hydrophobic-subtraction Model of Reversed-phase Column Selectivity, L.R. Snyder, J.W. Dolan and P.W. Carr, J. Chromatogr. A, 060 (004) 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)

7 hplc columns & accessories Application-Based Column Selection Use this chart to identify a suitable column for your application. Traditional choice columns are well matched to the application. Alternative choice columns can provide different selectivity when the traditional choice column does not work. Legend: Traditional Choice hplc columns Application-Based Column Selection Pharmaceutical Neutraceutical / Natural Products Food & Beverage Environmental Biopharmaceutical/ Life Science Acidic Column VisionHT C8 HighLoad VisionHT C8 Basic VisionHT C8 Classic VisionHT C8 Polar VisionHT Silica Alltima HP C8 Alltima HP C8 EPS Alltima HP C8 Hi-Load Alltima HP C8 AQ Alltima HP C8 Amide Alltima HP C8 Cyano Prevail C8 Select Prevail C8 Prevail Phenyl Prevail Organic Acid Prevail Carbohydrate ES GraceSmart C8 Vydac 8MS C8 Vydac 38MS C8 Vydac 08MS C8 Vydac 4MS C4 Vydac 9MS Diphenyl Everest 38EV C8 ProZap C8 Vydac 8TP C8 Vydac 4TP C4 *For UHPLC applications. Basic Neutral Herbal Medicines Vitamins Dietary Supplement Carbohydrates Lipids Organic Acids Food Safety Agricultural Chemicals Industrial/Aromatics Simple Enzymatic Digests (< proteins) Complex Enzymatic Digests (> proteins) Proteins, Peptides, Biomolecules 0 5K MW * * * * * * Alternative Choice 5 0K MW >0K MW Undenatured, Intact Proteins Antibodies Oligonucleotides 6

8 hplc columns & accessories Empty HPLC Hardware Stainless Steel Analytical Column Hardware Stainless Steel Preparative Column Hardware Threaded Column Tube Frit/Insert Endfitting Column Tube Continuously Adjustable Piston/Frit Endfitting hplc columns Stainless steel analytical columns come complete with the threaded tube, two µm industry standard frit/inserts, and two stainless steel endfitting nuts. Optional 0.5µm porosity frit/ inserts are also available. Adapters and sealing rings are available for slurry packing. The packing adapter comes with a /4" precolumn tube to connect to /4" slurry reservoirs. Stainless Steel Analytical Column Hardware Specifications Material: 36 Stainless Steel Max. Temperature: 00ºC Max. Pressure: 0,000psig Thread Type: 0-3 UNF (CPI Standard Ports) Typical Use: Analytical HPLC Columns Stainless Steel Analytical Hardware and Accessories.mm i.d. 4.6mm i.d. Complete Columns 50mm mm mm mm mm 6558 Replacement Parts Endfitting (ea) Frit/Insert, µm (5/pk) Frit/Insert, 0.5µm (5/pk) 750 The easy-to-use preparative format has threaded endfittings and a continuously adjustable piston/frit. The piston/frit extends column life by compressing the packed bed to remove voids. Stainless Steel Preparative Column Hardware Specifications Material: 36 Stainless Steel, PEEK Max. Temperature: 00ºC Max. Pressure: 8000psig Thread Type: 0-3 UNF (CPI Standard Ports) Typical Use: Prep HPLC Columns Stainless Steel Preparative Hardware and Accessories i.d. Length Qty. Complete Columns 7mm 00mm ea mm 50mm ea mm 00mm ea mm 50mm ea 965 mm 70mm ea 9654 mm 00mm ea 9650 mm 50mm ea 965 mm 500mm ea 965 Replacement Piston/Frit, µm 7mm ea mm ea 9657 mm ea 9657 Stainless Steel Frits Stainless Steel Frit Specifications Material: 36 Stainless Steel Max. Temperature: 500ºC Max. Pressure: 0,000psig Typical Use: Column Inlet/Outlet Stainless Steel Frits Frit Diameter Total Diameter Thickness Porosity 5/pk /4" (6.350mm) /4" (6.350mm) 0.03" (0.79mm) 0.5µm 7085 /4" (6.350mm) /4"(6.350mm) 0.03" (0.79mm).0µm 705 /4" (6.350mm) /4" (6.350mm) 0.063" (.59mm).0µm /8" (9.55mm) 3/8" (9.55mm) 0.03" (0.79mm).0µm 745 Encased Metal-Free Frits Encased Metal-Free PEEK Frit Specifications Material: PEEK Max. Temperature: 00ºC Max. Pressure: 0,000psig Typical Use: Column Inlet/Outlet Encased Metal-Free PEEK Frits Frit Diameter Total Diameter Thickness Porosity 5/pk PEEK Frits with PEEK Ring 0.8" (4.6mm) /4" (6.4mm) 0.063" (.59mm) 5µm " (4.6mm) /4" (6.4mm) 0.063" (.59mm) 0µm 6907 technical assistance Contact Tech Support: discoverysciences@grace.com Online: 7

9 hplc columns & accessories hplc columns VisionHT Columns One Platform for UHPLC, HPLC and PREP Easy method transfer between UHPLC, HPLC and preparative chromatography Broad selectivity range unique complementary phase chemistries Maximize speed and resolution with sub μm on any LC system Separate complex samples with excellent resolution with VisionHT high performance columns. They are available with.5, 3, 5 and 0μm particle sizes and six different phase selectivities spanning the full polarity spectrum. The same base silica and bonding chemistries are used across all particle sizes therefore offering simple method transfer between UHPLC, HPLC and preparative systems. VisionHT Phase Specifications Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties Phase C8 HighLoad 0 0 Yes Ultra-high purity silica, fully bonded. C8 Basic Proprietary Ultra-high purity silica. Controlled silica surface exposure gives dual mode separation with polar and non-polar analytes. C8 Classic Yes Lower carbon load. Slight silica exposure. C8 Polar No High silica exposure, low carbon load. Uniform coverage of inert vicinal silanols. HILIC 0 0 No Polar phase with shorter equilibration times. Shipped in ACN/Water. Silica 0 0 No Traditional normal phase for use in 00% organic mobile phases. Application/ Benefit General purpose for broad range compounds, classic selectivity, high-capacity for hydrophobic compounds. Alternate reversed-phase selectivity. High polar retention especially with compounds having two or more polar groups. Excellent sensitivity and peak shape for basic compounds, without the need for acidified mobile phases. Reversed-phase separations with reduced bonding optimized for speed. Some additional polar retention. Unique polar selectivity. Low carbon load gives fastest reversed-phase elution times while retaining 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. USP L-code L L L L L3 L3 Simple Method Transfer Optimizing and transferring methods between HPLC, UHPLC and preparative systems is not a simple and intuitive task. However, when the same base silica and bonding is available in sub-, 3, 5, and 0μm particles and in a variety of column formats, methods can be more easily transferred across system types and between laboratories, improving efficiency and productivity. Identical Selectivity Across All Particle Sizes Snyder-Dolan-Carr Coefficients for VisionHT C8 Basic Phase.5µm 3.0µm 5.0µm 0µm H S A B C.8 C7 References:. "The "Hydrophobic-subtraction" Model of Reversed-phase Column Selectivity", L.R. Snyder, J.W. Dolan and P.W. Carr, J. Chromatogr. A, 060 (004) "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) Method Transfer x 50mm, 6 5µm,.mL/min. 5. Uracil. Benzyl alcoho 3. Benzonitrile 4. Anisole 5. Ethyl benzene 6. Hexaphenone 4.6 x 50mm, 3µm,.mL/min. 7 x 53mm, 3µm,.0mL/min.. x 50mm,.5µm, 0.mL/min. 7 x 53mm, 3µm, 5.0mL/min min. Column: VisionHT C8, HighLoad, Dimensions noted Mobile Phase: A: 30mm phosphate buffer, ph7 B: Acetonitrile Gradient: Time: %B:

10 hplc columns & accessories Broad Selectivity Range Four reversed-phase column chemistries offer complementary selectivity. The unique VisionHT C8 Polar and Basic phases retain polar compounds while the C8 HighLoad has a more traditional reversed-phase selectivity. Using phases with orthogonal selectivity gives confidence that if one phase does not produce the separation the other will. VisionHT Reversed-Phases Span the Full Polarity Range VisionHT C8 Polar Phase VisionHT C8 VisionHT Basic C8 Column Basic Strongest retention for polar compounds Key VisionHT columns Commercially available columns hplc columns Increasing Polar Retention VisionHT C8 Classic Column Classic retention for hydrophobic compounds VisionHT C8 HighLoad High-Load Column Increasing Hydrophobic Retention References:. "The "Hydrophobic-subtraction" Model of Reversed-phase Column Selectivity", L.R. Snyder, J.W. Dolan and P.W. Carr, J. Chromatogr. A, 060 (004) "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) Different Selectivities of VisionHT Phases, Neutral Compounds. Benzonitrile. Anisole 3. 5,5-Diphenylhydantion 4. 5-Methyl salicylaldehyde VisionHT Classic VisionHT HighLoad VisionHT Basic VisionHT Polar min. Column: Various VisionHT Phases,.5µm,.0 x 50mm Mobile Phase: A: ACN B: Water Gradient: Time: 0 %B: 90 Flow Rate: 0.mL/min Detector: UV 0nm Different Selectivities of VisionHT Phases, Polar Compounds Acetaminophen. Caffeine 3. N,N'-Diethylacetamide 4. Benzonitrile min. Column: Various VisionHT Phases,.5µm,.0 x 50mm Mobile Phase: A: ACN B: Water Gradient: Time: 0 %B: 90 Flow Rate: 0.mL/min Detector: UV 0nm 4 VisionHT Classic VisionHT HighLoad VisionHT Polar VisionHT Basic 9

11 hplc columns & accessories Consistent and Reliable Performance Ultra-high mechanical strength silica and robust bonding chemistry minimize variations in capacity and selectivity. Media consistency combined with reproducible column packing methods delivers reliable performance and long column lifetime. hplc columns Reproducible Particle Technology Surface Area Mean = 07 RSD =.6% Pore Volume Mean = 0.5 RSD = 0.64% Pore Diameter Mean = 99.6 RSD =.5% 50 Consistent Column-to-Column Performance Biphenyl Nitrobenzene Amitriptyline Cinnamic Acid Toluene 6.00 Surface Area (m /g) Pore Diameter (Å) Pore Volume (cc/g) Capacity Factor Consecutive Batches Consecutive Batches VisionHT silica's surface area, pore volume, and pore diameter are highly reproducible. VisionHT media and advance packing methods produce columns with highly consistent capacity and selectivity. 0 0 Constant Performance after Routine Exposure to 6,000psig Pressure min. UHPLC columns rated to 6,000psig pressure limit Uracil (0.05mg/mL). Phenol (0.7mg/mL) 3. N,N-Diethyl-m-toluamide (0.6mg/mL) 4. Toluene (4.0mg/mL) INITIAL 3 4 min. 0 FINAL (After 0,000 column volumes at 6,000psig) Plates/Meter High-Pressure Stability Column Comparison, x 50mm* VisionHT Column VisionHT Column 5 VisionHT Column VisionHT Column 6 VisionHT Column 3 VisionHT Column 7 VisionHT Column 4 Competitor Sub µm Column Column Volumes Competitor Sub µm Column Column: Mobile Phase: Flow Rate: Detector: VisionHT C8 Polar,.5μm,.0 x 00mm ( 53960) (58:4) Acetonitrile:Water 0.mL/min UV at 0nm *After exposure to,000psig Before and after chromatograms show a consistent level of performance after exposure to 6,000psig pressures for 0,000 column volumes. Competitive columns lose performance over time under high-pressure conditions while VisionHT columns remain stable. 0

12 hplc columns & accessories Method Development Ultra MD Kits Experience Broad Selectivity Range of the VisionHT Phases Ultra MD kits include 4 phases with orthogonal selectivity to enable fast and efficient method development for the separation of very polar to very hydrophobic compounds. VisionHT Column Ultra MD Kits Phases Dimensions Ultra MD Kit C8 Classic, C8 Polar, x 00mm 5469 C8 HighLoad, C8 Basic Ultra MD Kit C8 Classic, C8 Polar, x 50mm C8 HighLoad, C8 Basic Ultra MD Kit 3 C8 Classic, C8 Polar, C8 HighLoad, C8 Basic x 50mm 5469 VisionHT Guard Columns Protect Your Column to Minimize Downtime and Reduce Cost Couple the zero-dead volume integral guard directly to any VisionHT column, with no loss in efficiency. hplc columns VisionHT Columns i.d. (mm) Length (mm) Particle Size (µm) C8 HighLoad C8 Basic C8 Classic C8 Polar HILIC Silica Format UHPLC Expedite Microbore Solvent Reducer Analytical Rocket Preparative Capillary Guard* All-Guard 3pk** *Requires Capillary holder, **Requires All-Guard Holder, 800. more info For detailed information on column formats, see page 3. technical assistance Contact Tech Support: discoverysciences@grace.com Online:

13 hplc columns & accessories hplc columns Alltima HP Columns Stability, Purity and Performance 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 Multiple selectivity options optimizes retention, resolution and analysis time Alltima HP columns combine our best phase chemistries with high-purity silica. The result is one product family with the selectivity and performance needed to help overcome your most challenging separation needs. Alltima HP Phase Specifications Phase Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties C Yes Classic reversed-phase retention and selectivity. Application/ Benefit Routine applications. C8 EPS Yes Greater retention and enhanced peak Reversed-phase applications symmetry for polar compounds. Alternate where C8 is too retentive. selectivity to traditional reversed-phases. C8 Hi-Load Yes Highest carbon load for superior retention High resolution for complex and loadability. samples. C8 AQ Yes 00% water wettable. Applications requiring high aqueous mobile phases. C8 Amide Yes Polar-embedded phase with extremely Basic compounds in neutral to low bleed. Compatible with microbore. alkaline ph, MS applications. C Yes Lower capacity compared to C8 phases. Reversed-phase applications where C8 is too retentive. Cyano Yes Extremely stable, long life and Ideal for basic drug analysis. reproducible. Silica No Highly polar phase. General purpose normal phase applications. HILIC 0 30 No Hydrophilic interaction chromatography uses small amounts of water for increased sensitivity with microbore applications. Very polar analytes that cannot be retained by reversed-phase. USP L-code L L L L L L7 L0 L3 L3 The Alltima HP Column Family Demonstrates Low Bleed Alltima HP columns are among the lowest-bleed columns available, making them ideal for microbore applications. The C8 amide polar-embedded phase and the cyano phase have significantly lower column bleed than competitive columns. Column Bleed by ESI-MS Alltima HP vs. Competitive Columns Alltima HP Cyano Much More Stable than Other Cyanos Relative Abundance Level (X0 5 ) No Column 0.8 Alltech Alltima HP Column Competitor Columns Alltima HP Silica Alltima HP C8 Alltima HP C8 Hi-Load C8 MS C8 MS.9 Alltima HP C8 Amide.5.8 C8 Amide 3. C8 Amide % Changes in Retention ph.0 ph 7.5 Alltima HP CN Brand A CN Brand B CN Brand C CN Brand D CN Brand E CN Brand F CN QC D4 Test: 70:30 HO/CH3CN,.0mL/min, UV at 54nm, 5µL D4Mix, Column 4.6 x 50mm, C ph.0 Stability: Flushed with HSO4 ph.0:acetonitrile 50:50 at.0ml/min for 3 days ph 7.5 Stability: Flushed with 0mM KHPO4 ph 7.5:Acetonitrile 50:50 at.0ml/min for 3 days

14 hplc columns & accessories Selectivity Options in One Product Family Capacity Factor k' Alltima HP Column Selectivity Comparisons CN k' Toluene k' Butyl Benzoic Acid k' Amitriptyline C8 EPS C8 C8 Amide C8 C8 AQ C8 Hi-Load Acid, Base and Neutral Test Mix Min. Column: 4.6 x 50mm Mobile Phase: 50mM KHPO4, ph 3.0:CH3CN (40:60) Flow Rate:.0mL/min Detector: UV at 54nm Uracil. Pyridine 3. Phenol 4. N,N-diethyl-m-toluamide 5. 4-Butyl Benzoic Acid 6. Toluene C8 C8 EPS C8 Hi-Load hplc columns Alltima HP Columns i.d. (mm) Length (mm) Particle Size (µm) C8 C8AQ EPS C8 C8 Hi-Load C8 Amide C8 CN Silica HILIC Format Capillary Expedite Microbore Solvent Reducer Analytical Rocket Guard*3pk *Requires All-Guard Holder, 800. more info For detailed information on column formats, see page 3. technical assistance Contact Tech Support: discoverysciences@grace.com Online: 3

15 hplc columns & accessories hplc columns Prevail Columns Stable from 00% Organic to 00% Aqueous Exceptional bonded phase stability for long column life Excellent sensitivity with microbore and ELSD applications Prevail HPLC columns offer exceptional versatility for difficult separations. Traditional reversed-phase packings experience bonded phase collapse under highly aqueous (>95%) conditions. Prevail bonded phases remain fully extended in 00% aqueous and offer a high carbon load for exceptional retention at 00% organic. Retain both polar and nonpolar compounds simply by adjusting the mobile phase. Prevail Phase Specifications Phase Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties Application/ Benefit C8 Select Yes Stable in 00% aqueous to Same benefits as Prevail C8, but slightly 00% organic mobile phases. more retentive. C Yes Stable in 00% aqueous to 00% organic mobile phases. Flexibility to switch between varied mobile phase conditions to suit a variety of applications. Excellent sensitivity for microbore applications. C Yes Stable C8 phase. Use for highly hydrophobic compounds that L7 retain too strongly on C8. Phenyl Yes Lowest hydrophobic capacity. Selective for aromatic compounds in a L variety of mobile phase conditions. Cyano Yes General purpose cyano Rugged normal phase applications. L0 suitable for normal or reversed-phase use. Amino No Stable in 00% aqueous to Use for carbohydrates or as a weak anion L8 00% organic mobile phases. exchanger. Silica No Highly polar phase. General purpose normal phase applications. L3 Organic Acid Yes Highly efficient silica-based, acid-stable. Separate common organic acids with unsurpassed resolution, speed and sensitivity. Lower cost than polymeric columns. Carbohydrate ES No Extremely stable hybrid phase. Excellent for mono- and oligosaccharides and sugar alcohols. USP L-code L L Switch Between 00% Aqueous and 00% Organic Mobile Phases on the Same Column Water-Soluble Vitamins (00% Aqueous) 9465 Fat-Soluble Vitamins and Tocopherols (00% Organic) 9405 Water-Soluble Vitamins (00% Aqueous) Thiamine. Ascorbic Acid 3. Nicotinic Acid 4. Pyridoxine 5. Niacinamide 6. Pantothenic Acid. Vitamin A. δ-tocopherol 3. γ-tocopherol 4. α-tocopherol (Vitamin E) 5. Vitamin D 6. Vitamin D3 7. Vitamin K Thiamine. Ascorbic Acid 3. Nicotinic Acid 4. Pyridoxine 5. Niacinamide 6. Pantothenic Acid wash with intermediate wash with intermediate Min Min Min. Column: Mobile Phase: Flow Rate: Detector: Prevail C8, 5µm, 4.6 x 50mm 5mM KHPO4 ph 3.0:CH3CN (97:3).0mL/min UV at nm Column: Prevail C8, 5µm, 4.6 x 50mm Mobile Phase: CH3CN:CH3OH (75:5) Flow Rate:.5mL/min Detector: UV at 0nm Column: Mobile Phase: Flow Rate: Detector: Prevail C8, 5µm, 4.6 x 50mm 5mM KHPO4 ph 3.0: CH3CN (97:3).0mL/min UV at nm 4

16 hplc columns & accessories Prevail Carbohydrate ES 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. 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. hplc columns Enhanced Detector Sensitivity Through Flat Gradient Baselines Min Min. Column: Prevail Carbohydrate ES, 5µm, 4.6 x 50mm Mobile Phase: A: Acetonitrile B: Water Gradient: Time: 0 5 %B: Flow Rate:.0mL/min Detector: ELSD. Dextrose. Maltose 3. Maltotriose 4. Maltotetraose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose 9. Maltononaose 0. Maltodecaose. Maltoundecaose. Maltododecaose 3. Maltotridecaose 4. Maltotetradecaose 5. Maltopentadecaose Robitussin Flu Syrup Min.. Sucralose. Fructose 3. Sorbitol 4. Glucose 5. Sucrose 6. Maltose Column: Prevail Carbohydrate ES, 5µm, 4.6 x 50mm Mobile Phase: A: Acetonitrile B: Water Gradient: Time: %B: Flow Rate:.0mL/min Detector: ELSD 0557 Prevail Columns i.d. (mm) Length (mm) Particle Size (µm) C8 C8 Select C8 Phenyl CN Amino Silica Organic Carbohydrate ES Format Expedite Microbore Solvent Reducer Analytical Rocket Guard* *Requires All-Guard Holder,

17 hplc columns & accessories hplc columns GraceSmart Columns High Quality at Exceptional Value High purity phases Efficient and reproducible separations Classic reversed-phase selectivity 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 for routine analysis or new method development, use GraceSmart columns to get premium performance at an exceptional value. GraceSmart Phase Specifications Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties Phase C Yes High purity silica with monomeric C8 bonding offers classical reversed-phase selectivity. Application/ Benefit Excellent value for general purpose, routine applications. USP L-code L Consistent and Reproducible Our tightly controlled silica synthesis and bonding keep capacity factor and selectivity variations to a minimum for acidic, basic, and neutral compounds. Competitive Performance GraceSmart columns show similar or better efficiency and asymmetries for challenging base and acid components in comparison to industry leading columns. Capacity Factor Toluene N-Butylbenzoic Acid Anisole Highly Reproducible Batch Acetophenone P-Nitrophenol Amitriptyline Symmetry Plates / Columns,000 0,000 8,000 6,000 4,000, Nicotine Nicotine Efficiency Quinine Symmetry Quinine Diphenhydramine Diphenhydramine Nortriptyline Nortriptyline Competitor C8 Column Competitor C8 Column,4 Dichlorophenoxy Acetic Acid,4 Dichlorophenoxy Acetic Acid,4 Dichlorophenoxy Propionic Acid,4 Dichlorophenoxy Propionic Acid,4 Dichlorophenoxy Butyric Acid,4 Dichlorophenoxy Butyric Acid Competitor 3 C8 Column GraceSmart C8 Column 4 3 Carbamates Min Fenuron. Monuron 3. Fluometuron 4. Diuron 5. Methiocarb 6. Linuron 7. Neburon 3 4 Folium Ginkgo Min Ginkgolide C. Bilobalide 3. Ginkgolide A 4. Ginkgolide B Column: GraceSmart C8, 5µm, 4.6 x 50mm Mobile Phase: A: 0.05M KHPO4, ph 3. B: Acetonitrile Gradient: Time: 0 0 %B: Flow Rate:.0mL/min Detector: UV at 40nm Column: GraceSmart C8, 5µm, 4.6mm x 50 Mobile Phase: Methanol:Tetrahydrofuran:Water (5:0:65) Flow Rate:.0mL/min Detector: ELSD 6

18 hplc columns & accessories mau Column: Mobile Phase: Flow Rate: Detector: Amoxicillin (U.S.P.) Min. GraceSmart C8 Column Luna C8 Column Inertsil ODS-3 Column Hypersil BDS C8 Column Kromasil 00-5 C8 Column 0697 GraceSmart C8, 5µm, 4.6 x 50mm Isocratic, 0.05N Monobasic Potassium Phosphate ph 5:Acetonitrile (96:4).5mL/min UV at 30nm Min. 7 Standards Mix. Nicotine. Quinine 3. Diphenhydramine 4. Nortriptyline 5.,4, Dichlorophenoxy Acetic Acid 6.,4, Dichlorophenoxy Propionic Acid 7.,4, Dichlorophenoxy Butyric Acid Column: GraceSmart C8, 5μm, 4.6 x 50mm Mobile Phase: 50% MeCN/50% 50mM, KHPO4, ph 3 Flow Rate:.0mL/min Detector: UV at 54nm Ibuprofen (U.S.P.) Valerophenone. Ibuprofen hplc columns Hypersil BDS C8 Kromasil 00-5 C8 USP GraceSmart Luna Inertsil Spec C8 C8 ODS-3 N/A Ret. Time (min) Theoretical plates Tailing Factor k'. k' Min. Column: GraceSmart C8, 5µm, 4.6 x 50mm Mobile Phase: HO (adjusted with Phosphoric Acid to a ph of.5): Acetonitrile (65:35) Flow Rate: ml/min Detector: UV at 4nm 3 4 Water Soluble Vitamins Min Ascorbic Acid. Nicotinic Acid 3. Pyridoxamine 4. Niacinamide 5. Folic Acid 6. Vitamin B 7. Riboflavin GraceSmart Columns Format i.d. (mm) Length (mm) Particle Size (µm) C8 Microbore Analytical Column: GraceSmart C8, 5μm, 4.6 x 50mm Mobile Phase: A: 0.M NH4HPO4 Buffer, ph 4.5 B: Acetonitrile Gradient: Time: %B: Flow Rate:.0mL/min Detector: UV at 54nm more info For detailed information on column formats, see page 3. technical assistance Contact Tech Support: discoverysciences@grace.com Online: 7

19 hplc columns & accessories hplc columns Vydac MS Columns Unsurpassed Resolution, Sensitivity and Recovery for Biomolecules 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 reversed-phase HPLC columns for biomolecules. A proprietary surface treatment and bonding process give Vydac MS columns truly unique selectivity. A variety of reversed-phases makes this product line suitable for small peptides to large intact, undenatured proteins. Vydac MS Phase Specifications Phase Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties 8MS C Yes Polymeric bonding, highest hydrophobic interaction and unique geometric selectivity. 38MS C Yes Monomeric bonding offers increased peptide interaction and generally yields higher peak counts. 08MS C Yes Lower hydrophobicity is better for larger biomolecules. 4MS C Yes Lower capacity than C8 or C8, suitable for hydrophobic proteins or when minimal organic is desired. 9MS Diphenyl Application/ Benefit Use for simple enzymatic digests (< proteins) or biomolecules 0 5K MW. Use for same applications as 8MS, but offers different C8 selectivity. Ideal for biomolecules 5 0K MW. Ideal for biomolecules >0K MW, undenatured intact proteins, antibodies, oligonucleotides, human growth hormone Yes Lowest capacity, aromatic functional groups. Highly selective for proteins with aromatic side chains. USP L-code L L L7 L6 L Unique Selectivity Improved selectivity for peptides on the Vydac MS columns results in better primary structure definition and easier identification of degradation products and other protein characteristics. Tryptic Digest of Fetuin Vydac 38MS C8 Column Competitor C8 Column Competitor C8 Column Min. Columns: All 300Å, 5µm, 4.6 x 50mm Mobile Phase: A: 0.% v/v trifluoroacetic acid in HO B: 0.085% v/v trifluoroacetic acid in Acetonitrile Gradient: Time: %B: Flow Rate:.0mL/min. Detector: UV at 5nm Sample: 30µL of a 6µg/µL digest (80µg total peptide load) Sample components interfere with the separation of peptides on the Competitor and columns, appearing as a chromatographic hump. The unique selectivity of Vydac MS columns solves separation problems. Unsurpassed Recovery and Peak Symmetry Many proteins are bound to the cell membrane and are very hydrophobic. Although hydrophobic proteins are particularly difficult to separate, Vydac MS columns provide excellent recovery and peak shape for these molecules. Transmembrane Protein p Min kd Protein. Non-myristolyated p4 3. Myristolyated p4 4. Triton X (Surfactant) Vydac 4MS C4 Column Competitor C4 Column Competitor C3 Column Competitor 3 C4 Column Competitor 4 C4 Column Columns: All 300Å, 4.6 x 50mm packed with 5µm except 4µm for Competitor Mobile Phase: A: 0.% v/v trifluoroacetic acid in HO B: 0.085% v/v trifluoroacetic acid in Acetonitrile Gradient: Time: %B: Flow Rate:.0mL/min. Detector: UV at 5nm Reptilian reovirus RRV p4 sample courtesy of Drs. Roberto J. de Antueno and Roy Duncan, Dalhousie University, Halifax, Nova Scotia. The Vydac column provides better performance for this highly hydrophobic membrane protein (RRV p4) and its fatty acid modified (myristolyated) form. 8

20 hplc columns & accessories High Protein Recovery Unique surface chemistry reduces adsorption of proteins for higher recoveries and increases mass loading, which makes Vydac MS columns ideal for preparative chromatography. Excellent Peak Shape with Little or No TFA TFA is UV absorbent and contributes background at low UV wavelengths. It is additionally problematic with electrospray MS where it interferes with ion generation and reduces MS sensitivity. Percent Recovery Vydac MS Column Loading Amount: µg of Cytochrome C Competitor Competitor Vydac MS C4 column provides more than 0% better recovery for Cytochrome C. Vydac MS Columns Competitive Recovery Comparisons i.d. (mm) Length (mm) Particle Size (µm) 8MS (C8) 38MS (C8) TFA Concentration Comparisons on 8MS Columns 0.6 AU % TFA 0.0% TFA 0.0% TFA 08MS (C8) Min. 4MS (C4) 4 9MS (diphenyl). Neurotensin 8 (fragment). Oxytocin 3. Angiotensin II 4. Neurotensin Format Capillary MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS MS5.50 4MS MS MS MS5.55 4MS MS MS MS5.55 4MS MS MS MS MS MS MS MS5.30 4MS MS MS MS5.35 4MS MS MS MS5.35 4MS MS MS MS MS MS MS MS5.50 4MS MS MS MS5.55 4MS MS MS MS5.55 4MS5.55 Microbore MS505 38MS505 08MS505 4MS MS50 38MS50 08MS50 4MS50 9MS MS55 38MS55 08MS55 4MS MS5 38MS5 08MS5 4MS5 Holder Solvent MS53 4MS53 Reducer Analytical MS5405 4MS MS540 38MS540 4MS MS545 38MS545 08MS545 4MS545 9MS MS54 38MS54 08MS54 4MS54 Preparative MS MS50 4MS MS00 4MS MS05 4MS MS0 4MS0 Guard GD5MS 38GD5MS 08GD5MS 4GD5MS GD54MS 38GD54MS 08GD54MS 4GD54MS 800 Prep Guard GCC50 4GCC GCC GCC530 4GCC hplc columns 9

21 hplc columns & accessories hplc columns Vydac Everest Columns High Resolution for Complex Peptide Digests Exceptionally high peak counts for better characterization Unique selectivity for both hydrophilic and hydrophobic peptides Excellent sensitivity with little or no TFA in the mobile phase Everest columns have unique selectivity and sensitivity, which are the result of bonding technology that improves C8 surface coverage and deactivates residual silanols. Previously, the best 300Å C8 chemistries have had carbon coverage in the.8 to 3.6µmol/m range. Everest C8 coverage is in excess of 4µmol/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. Vydac Everest Phase Specifications Pore Size (Å) Surface Area (m /g) Carbon Load (%) Endcapped Chromatographic Properties Phase 38EV C Yes Maximum surface coverage for highest resolution of complex samples. Application/ Benefit Complex enzymatic digests (> proteins). USP L-code L Tryptic Digest of Bovine Serum Albumin Everest Column 0 peaks Commercial Column A 94 peaks Commercial Column B 96 peaks Min. Column: All C8, 300Å, 5µm, 4.6 x 50mm Mobile Phase: A: 0.% TFA in Water B: 0.085% TFA in Acetonitrile Gradient: Time: %B: Flow Rate:.0mL/min Detector: UV at 5nm Everest columns outperform competitor columns by providing higher-resolution separations (average of 7% higher peak counts than competitor columns tested). Vydac Everest Columns i.d. Length Particle Size 38EV (C8) Format (mm) (mm) (µm) Capillary EV EV EV EV EV EV EV EV EV EV EV5.55 Microbore EV EV EV EV EV EV5 Analytical EV EV EV54 Bovine Serum Albumin with no TFA Tryptic Digest Intensity (x 0 6 cps) Min. Column: Vydac Everest C8, 300Å, 5µm, 0.3 x 50mm Mobile Phase: A: 0.% v/v Formic Acid in Water B: 0.% v/v Formic Acid in Acetonitrile Gradient: Time: %B: Flow Rate: 5µL/min Detector: MS + ESI The Everest column performs exceptionally well with no TFA in the mobile phase, ensuring excellent microbore sensitivity. Vydac Everest Guard Columns i.d. (mm) Length (mm) Particle Size (µm) 38EV (C8) Holder Format Capillary EV5C030 GR-370A Guard Guard GD5EV GD54EV 800 more info For detailed information on column formats, see page 3. technical assistance Contact Tech Support: discoverysciences@grace.com Online: 0