Preparative HPLC columns Eurospher II for semipreparative and preparative HPLC

Transcription

1 Preparative HPLC columns Eurospher II for semipreparative and preparative HPLC Preparative column hardware with axial compression -> Longevity Wide range of modifications available -> Flexibility Extensive application services -> Column and method consultation KNAUER Wissenschaftliche Geräte GmbH Hegauer Weg Berlin

2 Vertex Plus AX Preparative KNAUER column hardware with axial compression What is the technical principle behind Vertex Plus AX? Up to 30% (25-40 mm) of the column bed length can be readjusted via the adjustable column screw. What characterizes the Vertex Plus AX life time? 1) Column using with highest packing quality 2) readjustment of the Vertex Plus AX column bed after demanding conditions 3) reconstituted column performance for a longer life time Example for Column Readjustment of a 250 x 20 mm Vertex Plus AX column: Cost-effective Taking the features and advantages of the KNAUER Vertex Plus AX preparative HPLC columns into account, we offer a highly competitive product. Available column dimensions**: The upper limit for the column backpressure depends on the inner diameter of the column and is specified as 400 bar for 8,16 and 20 mm ID, 300 bar for 30 mm ID and 200 bar for 50 mm ID. ** analytical column dimensions from 2 mm up to 4,6 mm ID also available

3 Eurospher II Family Based on an ultra pure spherical silica gel, Eurospher II is a high performance column material for analytical, semipreparative and process-scale applications. Eurospher II features very narrow particle and pore size distributions, as well as outstanding mechanical stability. Eurospher II silica gel is perfectly suited to take on preparative separations of varying substances. Technical Data: Silica gel ultrapure > % Metal content < 10 ppm Particle size 5 μm, 10 μm, 15 μm, μm Particle shape spherical Porengröße 100 Å Specific surface 320 ± 20 m2/g Pore volume 0.8 ml/g Density 430 g/l * analytical columns also available with 2 and 3 µm particle size Characteristics: Eurospher II 100 HPLC phases are based on ultra pure 3rd generation silica gel numerous modifications are available for a broad range of applications high resolution LC phase, suited for analytical HPLC and UHPLC as well as for semipreparative and preparative Separations high mechanical stability narrow particle and pore size distribution high batch to batch reproducibility Recommended application areas:

4 Characterisation of phases based on Tanaka tests Optimized selectivity for better separations HR: HS: SS: HBC: hydrophobic retention hydrophobic selectivity steric selectivity hydrogen bonding capacity IEX (ph 7,6): ion exchange capacity at ph 7.6 IEX (ph 2,7): ion exchange capacity at ph 2.7 Eurospher II 100 C18 - USP L1 - for acidic, basic and neutral analytes in reversed phase mode (sulphonamides; anabolic steroids; anti-psychotics; beta blocker; Sudan dyes; phenols, preservatives etc.) Eurospher II 100 C18 A - USP L1- polar endcapped C18 phase for alternative selectivity; 100% aqueous applications with very polar compounds (basic pharmaceutical ingredients, water soluble vitamins, catecholamines as well as organic acids) Eurospher II 100 C18 H - USP L1 - recommended alternative for Kromasil 100 C18 columns; for acidic, basic and neutral analytes in reversed phase mode with extended ph range

5 Eurospher II 100 C18 P - USP L1 - alternative selectivity to C18 phase; stationary phase in Eurospher II C18 family with the highest carbon load; fully endcapped; excellent shape selectivity and ph stability Eurospher II 100 C8 - USP L7 - similar selectivity to C18 phase but less retention due to the lower hydrophobicity; useful for determination of water soluble vitamins, steroids, catecholamines, sedatives etc. Eurospher II 100 Phenyl - USP L11 - alternative selectivity for aromatic and mode rately polar analytes or mixtures with varying polarity and aromaticity NH2 Eurospher II 100 NH2- USP L8 - most flexible phase in the Eurospher II family; can be used in three modes: normal phase, reversed phase and ion chromatography mode (weak anion exchanger); different selectivity to the silica packing; in reversed phase mode mainly used for analysis of carbohydrates

6 Eurospher II 100 C18 - USP L1 - Ultra pure, spherical high performance HPLC phase based on silica gel Unpolar, monomeric C18 (Octadecyl) modification, endcapped, 16 % carbon content (~ 50 % endcapping). Properties: Separation mechanism: Hydrophobic interaction Key features: High-class HPLC phase perfectly suited to take on routine analyses as well as the most ambitious chromatography tasks, classical C18 phase with ca. 50 % endcapping and resulting 16 % carbon content, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical data: C18 phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: Suitable for acidic, basic and neutral analytes in reversed phase mode, for example sulphonamides; anabolic steroids; anti-psychotics; beta blocker; sudan dyes; phenols and preservatives Eurospher II C18, 150 x 30 mm Article number: 15QE181E2N, Application code: VEV0062J 1 Ethyl acetate 100 mg/ml 2 Butyl acetate 100 mg/ml 3 Pentyl acetate 100 mg/ml Injection volume 1 ml Never use classical C18 phases with 100 % aqueous mobile phase. The hydrophobic C18 chains will collapse. Only special C18 A or C18 AQ phases have a hydrophilic endcapping style and can be used with pure water. Eurospher II 100 C18 article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE181E2J 25PE181E2N 25PE181E2Q 25QE181E2J 25QE181E2N 25QE181E2Q 25RE181E2N 25RE181E2Q 25RE181E2X 150 mm 15PE181E2J 15PE181E2N 15PE181E2Q 15QE181E2J 15QE181E2N 15QE181E2Q 15RE181E2N 15RE181E2Q 15RE181E2X 30 mm precolumn 03PE181E2J 03PE181E2N 03PE181E2Q 03PE181E2J 03PE181E2N 03PE181E2Q 03RE181E2N 03RE181E2Q 03RE181E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE181E2J 30GE181E2N 10 g 00BE181E2J 00BE181E2N 00BE181E2Q 00BE181E2X 250 mm 25GE181E2J 25GE181E2N 25IE181E2J 25IE181E2N 100 g 00CE181E2J 00CE181E2N 00CE181E2Q 00CE181E2X 125 mm 12GE181E2J 12GE181E2N 12IE181E2J 12IE181E2N 50 mm 05GE181E2J 05GE181E2N 05IE181E2J 05IE181E2N 30 mm 03GE181E2J 03GE181E2N 03IE181E2J 03IE181E2N other column dimensions and particle sizes upon request, please check

7 Eurospher II 100 C18 A - USP L1- Ultra pure, spherical high performance HPLC phase based on silica gel Polar, monomeric C18 (Octadecyl) modification, polar endcapping, 10 % carbon content (~ 50 % endcapping) Properties: Separation mechanism: Hydrophobic and polar interaction Key features: C18 A phase with alternative, more polar selectivity, suited for the use with pure aqueous mobile phases, 50 % endcapping and resulting 10 % carbon content, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical data: C18 A phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: polar endcapped C18 phase for water soluble and polar analytes, 100 % aqueous eluents for analysis of very polar compounds, basic pharmaceutical ingredients, water soluble vitamins, catecholamines as well as organic acids. Eurospher II C18, 150 x 20 mm ID Bestellnummer: 15PE181E2N; Application code: VFD0106J 1 trans-2-hexen-1-al 2 trans-2-hexen-1-ol 3 Methyl 2-methylbutyrate; Ethyl isobutyrate; Ethyl butyrate; n-butyl acetate 4 Ethyl 2-methylbutyrate;Ethyl valerate; 3-Methylbutyl acetate; 2-Methylbutyl acetate 5 other compounds (elution with higher Ethanol concentration) Injection voume blue 0,1 ml and green 1 ml apple extract Before a preparative purification always an analytical method delopment has to be done to find the best method paramters like nature of the eluent and stationary phase. For the final scale up, the method should be optimized regarding productivity Eurospher II 100 C18 A article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE184E2J 25PE184E2N 25PE184E2Q 25QE184E2J 25QE184E2N 25QE184E2Q 25RE184E2N 25RE184E2Q 25RE184E2X 150 mm 15PE184E2J 15PE184E2N 15PE184E2Q 15QE184E2J 15QE184E2N 15QE184E2Q 15RE184E2N 15RE184E2Q 15RE184E2X 30 mm precolumn 03PE184E2J 03PE184E2N 03PE184E2Q 03PE184E2J 03PE184E2N 03PE184E2Q 03RE184E2N 03RE184E2Q 03RE184E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE184E2J 30GE184E2N 10 g 00BE184E2J 00BE184E2N 00BE184E2Q 00BE184E2X 250 mm 25GE184E2J 25GE184E2N 25IE184E2J 25IE184E2N 100 g 00CE184E2J 00CE184E2N 00CE184E2Q 00CE184E2X 125 mm 12GE184E2J 12GE184E2N 12IE184E2J 12IE184E2N 50 mm 05GE184E2J 05GE184E2N 05IE184E2J 05IE184E2N 30 mm 03GE184E2J 03GE184E2N 03IE184E2J 03IE184E2N other column dimensions and particle sizes upon request, please check

8 Eurospher II 100 C18 H - USP L1 - Ultra pure, spherical high performance HPLC phase based on silica gel Unpolar, monomeric C18 (Octadecyl) modification, high efficiency endcapping, 17 % carbon content (~ 99 % endcapping) Properties Separation mechanism: Hydrophobic interaction Key features: Unpolar C18 phase with high efficiency endcapping and 17 % carbon content, therewith higher ph stability in the range of 1-12, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical data: C18 phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: Recommended alternative to Kromasil 100 C18, für acidic, basic and neutral substances in reversed phase mode with extended ph range Eurospher II C18, 250 x 20 mm ID with precolumn 30 x 20 mm ID Bestellnummer: 25PE181E2J, 03PE181E2J, Application code: VFD0118J 1 Piperin Injection volume 1 ml pepper extract Most silica gel based phases are only stable at ph 2-8. Eurospher II 100 C18 H and C18 P are exceptions that are stable at ph 1-12 Eurospher II 100 C18 H article numbers Preparative columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE185E2J 25PE185E2N 25PE185E2Q 25QE185E2J 25QE185E2N 25QE185E2Q 25RE185E2N 25RE185E2Q 25RE185E2X 150 mm 15PE185E2J 15PE185E2N 15PE185E2Q 15QE185E2J 15QE185E2N 15QE185E2Q 15RE185E2N 15RE185E2Q 15RE185E2X 30 mm precolumn 03PE185E2J 03PE185E2N 03PE185E2Q 03PE185E2J 03PE185E2N 03PE185E2Q 03RE185E2N 03RE185E2Q 03RE185E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE185E2J 30GE185E2N 10 g 00BE185E2J 00BE185E2N 00BE185E2Q 00BE185E2X 250 mm 25GE185E2J 25GE185E2N 25IE185E2J 25IE185E2N 100 g 00CE185E2J 00CE185E2N 00CE185E2Q 00CE185E2X 125 mm 12GE185E2J 12GE185E2N 12IE185E2J 12IE185E2N 50 mm 05GE185E2J 05GE185E2N 05IE185E2J 05IE185E2N 30 mm 03GE185E2J 03GE185E2N 03IE185E2J 03IE185E2N other column dimensions and particle sizes upon request, please check

9 Eurospher II 100 C18 P - USP L1 - Ultra pure, spherical high performance HPLC phase based on silica gel Unpolar, trifunctional C18 (Octadecyl) modification, high efficiency endcapping 20 % carbon content (~ 99 % endcapping) Properties Separation mechanism: Hydrophobic and steric interaction Key features: KNAUER s most unpolar C18 phase, polymeric, high efficiency endcapping of 99 % and 20 % carbon content, higher ph stability in the range of 1-12, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical data: C18 phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: alternative selectivity to standard C18; stationary phase in Eurospher II C18 family with the highest carbon load; fully endcapped; excellent shape selectivity and ph stability Method development part 1: Not all C18 phases are comparable When replacing an existing column always have a close look at the characteristics like carbon load, pore size and specific surface area Eurospher II C18, 30 x 50 mm ID Article number: 03RE181E2N, Application code: VFD0108J 1 Sudan II 2 mg/ml 2 Sudan III 2 mg/ml Injection volume 5 ml If your samples are not complex, it is worth switching to larger particle sizes (10 µm or bigger) and reducing therewith backpressure and column price. Eurospher II 100 C18 P article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE182E2J 25PE182E2N 25PE182E2Q 25QE182E2J 25QE182E2N 25QE182E2Q 25RE182E2N 25RE182E2Q 25RE182E2X 150 mm 15PE182E2J 15PE182E2N 15PE182E2Q 15QE182E2J 15QE182E2N 15QE182E2Q 15RE182E2N 15RE182E2Q 15RE182E2X 30 mm precolumn 03PE182E2J 03PE182E2N 03PE182E2Q 03PE182E2J 03PE182E2N 03PE182E2Q 03RE182E2N 03RE182E2Q 03RE182E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE182E2J 30GE182E2N 10 g 00BE182E2J 00BE182E2N 00BE182E2Q 00BE182E2X 250 mm 25GE182E2J 25GE182E2N 25IE182E2J 25IE182E2N 100 g 00CE182E2J 00CE182E2N 00CE182E2Q 00CE182E2X 125 mm 12GE182E2J 12GE182E2N 12IE182E2J 12IE182E2N 50 mm 05GE182E2J 05GE182E2N 05IE182E2J 05IE182E2N 30 mm 03GE182E2J 03GE182E2N 03IE182E2J 03IE182E2N other column dimensions and particle sizes upon request, please check

10 Eurospher II 100 C8 - USP L7 - Ultra pure, spherical high performance HPLC phase based on silica gel Monomeric C8 (Octyl) modification, standard endcapping 10 % carbon content (~ 50 % endcapping) Properties Separation mechanism: Hydrophobic interaction (lower compared to C18 phases) Key features: Classical C8 phase, standard 50 % endcapping and 10 % carbon load, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical data: C8 phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: Similar selectivity to C18 phase but less retention due to the lower hydrophobicity; useful for determination of water soluble vitamins, steroids, catecholamines, sedatives etc. Eurospher C18, 150 x 30 mm ID Article number: 15QE181E2N, Application code: VPH0059J Method development part 2: The choice of the mobile phase is very important when optimizing a preparative HPLC method. At the same time, viscosity (backpressure) and costs should be regarded. In the end youir mobile phase should also be easy to evaporate and separate from your target product. 1 Cortisone 5 mg/ml 2 Corticosterone 5 mg/ml 3 Desoxycorticosterone 5 mg/ml Injection volume 1 ml Eurospher II 100 C8 article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE081E2J 25PE081E2N 25PE081E2Q 25QE081E2J 25QE081E2N 25QE081E2Q 25RE081E2N 25RE081E2Q 25RE081E2X 150 mm 15PE081E2J 15PE081E2N 15PE081E2Q 15QE081E2J 15QE081E2N 15QE081E2Q 15RE081E2N 15RE081E2Q 15RE081E2X 30 mm precolumn 03PE081E2J 03PE081E2N 03PE081E2Q 03PE081E2J 03PE081E2N 03PE081E2Q 03RE081E2N 03RE081E2Q 03RE081E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 Bulk Media Column length 5 µm 10 µm mm 5 µm ID 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE081E2J 30GE081E2N 10 g 00BE081E2J 00BE081E2N 00BE081E2Q 00BE081E2X 250 mm 25GE081E2J 25GE081E2N 25IE081E2J 25IE081E2N 100 g 00CE081E2J 00CE081E2N 00CE081E2Q 00CE081E2X 125 mm 12GE081E2J 12GE081E2N 12IE081E2J 12IE081E2N 50 mm 05GE081E2J 05GE081E2N 05IE081E2J 05IE081E2N 30 mm 03GE081E2J 03GE081E2N 03IE081E2J 03IE081E2N other column dimensions and particle sizes upon request, please check

11 Eurospher II 100 Phenyl - USP L11 - Ultra pure, spherical high performance HPLC phase based on silica gel Phenyl modification (Phenylpropyl), 12 % carbon content Properties Separation mechanism: Pi-Pi Interactions Key features: Classical phenyl phase with 12 % carbon load, outstanding mechanical and chemical stability, well suited for analytical, semi preparative and preparative separation tasks Technical Data: Phenyl phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 2 and 3 μm particle size) Recommended application areas: Alternative selectivity for aromatic and mode rately pi-pi interaction polar analytes or mixtures with varying polarity and aromaticity Empfohlene Anwendungsgebiete: Alternative Selektivität für aromatische und moderat polare Analyten, besonders mit Pi-Pi-Bindungen oder Mischungen mit variierender Aromatizität. Method development part 3: Whenever possible, isocratic runs are recommended to simplify the scale up process. For more complex methods KNAUER provides the KNAUER ScaleUp Converter tool. Check for free Eurospher II C18, 150 x 4 mm ID und 150 x 20 mm ID Article number: 15DE181E2N und 15PE181E2N, Application code: VFD0107J green: analytical 4 mm ID, 10 µl blue: preparative 20 mm ID, 250 µl Eurospher II 100 Phenyl article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE050E2J 25PE050E2N 25PE050E2Q 25QE050E2J 25QE050E2N 25QE050E2Q 25RE050E2N 25RE050E2Q 25RE050E2X 150 mm 15PE050E2J 15PE050E2N 15PE050E2Q 15QE050E2J 15QE050E2N 15QE050E2Q 15RE050E2N 15RE050E2Q 15RE050E2X 30 mm precolumn 03PE050E2J 03PE050E2N 03PE050E2Q 03PE050E2J 03PE050E2N 03PE050E2Q 03RE050E2N 03RE050E2Q 03RE050E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE050E2J 30GE050E2N 10 g 00BE050E2J 00BE050E2N 00BE050E2Q 00BE050E2X 250 mm 25GE050E2J 25GE050E2N 25IE050E2J 25IE050E2N 100 g 00CE050E2J 00CE050E2N 00CE050E2Q 00CE050E2X 125 mm 12GE050E2J 12GE050E2N 12IE050E2J 12IE050E2N 50 mm 05GE050E2J 05GE050E2N 05IE050E2J 05IE050E2N 30 mm 03GE050E2J 03GE050E2N 03IE050E2J 03IE050E2N other column dimensions and particle sizes upon request, please check

12 Eurospher II 100 NH2 - USP L8 - Ultra pure, spherical high performance HPLC phase based on silica gel Amino modification (Aminopropyl), 4 % carbon content (without endcapping) Properties Separation mechanism: Hydrophilic and ionic interactions Key features: Multi mode column for RP, NP, HILIC and IC, Aminopropyl modification without endcapping, outstanding mechanical stability, well suited for analytical, semi preparative and preparative separation tasks Technical Data: Amino phase based on ultra-pure silica gel (purity > 99 %), metal content < 10 ppm, ph-stability 2-8, pore size 100 A, particle size 5 μm, 10 μm, 15 μm, μm (analytical columns also available in 3 μm particle size) Recommended application areas: Most flexible phase in the Eurospher II family; can be used in three modes: normal phase, reversed phase and ion chromatography mode (weak anion exchanger); different selectivity to the silica packing; in reversed phase mode mainly used for analysis of carbohydrates Eurospher II NH2, 100 x 3 mm ID Article number: 10CE190E2G, Application code: VFD0089J NH2 columns, that are used in normal phase mode should be stored in an inert solvent like heptane or hexane for example. When used in reversed phase mode the water content should be as low as possible for column storage. Best would be applying pure acetonotrile because NH2 phases are highly sensitive to water. 1 Fructose 2 Glucose 3 Saccharose 4 Maltose 5 Maltotriose Eurospher II 100 NH2 article numbers Columns Vertex Plus AX 20 mm ID Vertex Plus AX 30 mm ID Vertex Plus AX 50 mm ID Column length 5 µm 10 µm 15 µm 5 µm 10 µm 15 µm 10 µm 15 µm µm 250 mm 25PE190E2J 25PE190E2N 25PE190E2Q 25QE190E2J 25QE190E2N 25QE190E2Q 25RE190E2N 25RE190E2Q 25RE190E2X 150 mm 15PE190E2J 15PE190E2N 15PE190E2Q 15QE190E2J 15QE190E2N 15QE190E2Q 15RE190E2N 15RE190E2Q 15RE190E2X 30 mm precolumn 03PE190E2J 03PE190E2N 03PE190E2Q 03PE190E2J 03PE190E2N 03PE190E2Q 03RE190E2N 03RE190E2Q 03RE190E2X Semi-preparative Vertex Plus 8 mm ID Vertex Plus 16 mm ID Bulk Media Column length 5 µm 10 µm 5 µm 10 µm Quantity 5 µm 10 µm 15 µm µm 300 mm 30GE190E2J 30GE190E2N 10 g 00BE190E2J 00BE190E2N 00BE190E2Q 00BE190E2X 250 mm 25GE190E2J 25GE190E2N 25IE190E2J 25IE190E2N 100 g 00CE190E2J 00CE190E2N 00CE190E2Q 00CE190E2X 125 mm 12GE190E2J 12GE190E2N 12IE190E2J 12IE190E2N 50 mm 05GE190E2J 05GE190E2N 05IE190E2J 05IE190E2N 30 mm 03GE190E2J 03GE190E2N 03IE190E2J 03IE190E2N other column dimensions and particle sizes upon request, please check

13 Technical Documentation Vertex Plus AX- preparative KNAUER columns with axial compression How satisfied are you with your current preparative column performance and reproducibility? How long is your preparative column life time? What are Vertex Plus AX columns? The new preparative columns from KNAUER are a product of choice for every HPLC user who needs highest performance as well as robust and long lasting columns in preparative range. The column hardware realizes axial compression of the column bed to get long lasting columns with excellent performance. Which column dimensions are available? The new preparative column hardware can be offered with three different internal diameters: - 20 mm ID with 1/16 capillary - 30 mm ID with 1/8 capillary - 50 mm ID with 1/8 capillary The Vertex Plus AX column length will start with a - 30 mm precolumn mm and mm column length is the typical bed length for the main column The easiest way to store your column safdely during measurements or for storage is the prism column holder What are the advantages of Vertex Plus AX? preparative HPLC columns with adjustable end fitting consistent performance for extended time period by readjustment of column fitting optimum packed bed density and stability in a universal design available with KNAUER and NON-KNAUER column materials flexibility in column design makes it suitable for any media What are the preparative working parameters? Column dimension Column volume [ml] Scale up factor* Loadability** [mg] Recommended Flow rate [ml/min] 150 x 20 mm ID 33 18,9 up to x 20 mm ID 55 18,9 up to x 30 mm ID 79 42,5 up to x 30 mm ID ,5 up to x 50 mm ID up to x 50 mm ID up to *Reference 4.6 mm ID **Loadability highly depends on the characteristics of the sample, only indications are given in the table

14 Technical Documentation Equilibration, regeneration and storage of silica gel based columns like Eurospher II Proper storage of silica based HPLC columns Silica based columns should be stored in an aprotic solvent. The best solvent for storage of RP packings (C18, C8, C4, C1, C30, CN and Phenyl) is acetonitrile/water (50:50 v/v). The water content should not be greater than 50%. The best solvent for storage of NP packings (Silica, Diol, Nitro, Cyano and Amino) is hexan/isopropanol 90:10 (v/v). The best solvent for storage of columns used in HILIC mode (HILIC, Amino, and Silica) is acetonitrile/water (90:10 v/v) or acetonitrile/5 mm ammonium acetate, ph 5,3 (90:10 v/v). Acetonitrile content should always be greater than 90 %. Caution Even for short-term storage, flush out all buffer solution from the column to prevent algal growth. Make sure that all buffers are washed out of the column before exchanging aqueous mobile phases by organic solvents. Buffer salts are not soluble in acetonitrile and can block capillary tubing and the column. Equilibration The equilibration time of a column depends on the column dimensions. In general, a column is equilibrated after 20 column volumes are flushed through it. The equilibration times for the most important column dimensions are summarized in the following table. You can reduce the equilibration time by simply increasing the flow rate. However, make sure to flush the column with at least column volumes to make sure the column is equilibrated. Column dimension (Length x ID) Column volume [ml] Flow rate [ml/min] Equilibration time [min] maximum backpressure [bar] 250 x 8 mm x 8 mm x 8 mm x 16 mm x 16 mm x 16 mm x 20 mm x 20 mm x 30 mm x 30 mm x 50 mm x 50 mm Did you ever wonder how to maintain the temperature of a preparative HPLC column? We recommend to use heating sleeves and optional for higher flow rates eluent preheating modules.

15 Technical Documentation Equilibration, regeneration and storage of silica gel based columns like Eurospher II Regeneration of a column We recommend regenerating a column if a change in peak form, retention time, resolution or an increase in backpressure is observed. If the system pressure begins to rise, remove the column and check the system to find whether the pressure increase is being caused by the system or the column. Pressure increase caused by system: flush system, exchange eluent filters, frits and/or blocked capillaries. Pressure increase caused by column: backflush the column carefully to remove particle buildup from the inlet frit (connect the column outlet to the pump/injector and flush). Do not connect the column to the detector. If the column still has a high backpressure, flush the column according the following regeneration scheme: Regeneration scheme for RP columns (C18, C8, C4, C1, C30, CN and Phenyl stationary phases) Regeneration scheme for NP columns (Silica, Diol, Nitro, Cyano and Amino stationary phases) Regeneration scheme for columns used in HILIC mode (HILIC and Silica stationary phases) 20 column volumes water 20 column volumes heptane 20 column volumes water 20 column volumes acetonitrile 5 column volumes isopropanol 30 column volumes 0.5 M ammonium acetate 5 column volumes isopropanol 20 column volumes acetonitrile 30 column volumes water 20 column volumes heptane 20 column volumes water 20 column volumes acetonitrile/water (50:50 v/v) 5 column volumes isopropanol 20 column volumes acetonitrile 20 column volumes acetonitrile 20 column volumes acetonitrile 5 column volumes isopropanol 20 column volumes acetonitrile/water (50:50 v/v) 20 column volumes heptane After the regeneration procedure, re-equilibrate the column with the mobile phase before analysis. Do you need help with method upscaling to the preparative scale? Our LC LAB team would be happy to assist you. All LC LAB services can be found online, please check

16 HPLC SMB Osmometry Wide range of applications and columns on HPLC SMB Osmometry KNAUER Wissenschaftliche Geräte GmbH Hegauer Weg Berlin, Germany Internet: Phone: Telefax: *Terms, technical data and prices are subject to change without notice. Visit terms.html for complete Terms and Conditions. V7804-1US/2.0/04.16/MM/Lo