Eurospher II the logical choice. Physical properties of Eurospher II silica gel: Silica gel: ultra pure, > % Metal content: < 10 ppm

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1 HPLC SMB smometry Euroline Eurospher II Eurospher II the logical choice After more than years on the market, our Eurospher stationary phase has established a reputation for being a first-class packing material for a wide field of applications. ur next generation Eurospher II has been developed for an even wider range of application areas. 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 routine analyses as well as the most ambitious chromatography tasks. Physical properties of Eurospher II silica gel: Silica gel: ultra pure, > % Metal content: < 1 ppm Particle size: µm, µm, 1 µm, (1 µm, µm upon request) Particle shape: spherical Pore size: 1 Å Specific surface: ± m /g Pore volume:.8 ml/g Density: g/l Eurospher II offers outstanding mechanical and chemical stability. With physical properties very similar to those of Kromasil 1, Eurospher II columns can be used to replace Kromasil columns, providing excellent peak symmetry for acids, bases, and neutrals.

2 HPLC SMB smometry Eurospher II is comparable with Kromasil and even outperforms it in some respects: nearly the same particle shape as Kromasil 1 Eurospher II has a lower metal impurity specification higher mechanical stability compared to Kromasil 1 comparable selectivity in P mode of Eurospher II C18 H and Kromasil 1 C18 Modifications With the wide range of different surface modifications available, all application fields in reversed phase and normal phase modes are covered. Every Eurospher II modification offers high chemical stability and loading capacity thanks to mono- and multi-functional silanes. With several different levels of endcapping, Eurospher II offers a wide variety of surface modification types for analytical as well as preparative columns. ur long experience and knowledge in producing HPLC columns ensures the highest reproducibility. Standard liquid endcapping (~ %) P P P H H H H H Si Si Si Si Si Si Si Si Si Si Si Si P: C 8 or C 18 chain High efficiency endcapping (~ 99 %) P P P Si Si Si Si Si Si Si Si Si Si Si Si In summary four different Eurospher II C18 types are now available. Eurospher II reversed phase columns are up to the challenge by delivering outstanding selectivity of positional isomers, steric isomers, and polar compounds. Modification USP code % carbon ph range C18 L1 1 % (~ % endcapping) 8 C18 H L1 17 % (> 99 % endcapping) 1 1 C18 P L1 % (~ 99 % endcapping) 1 1 C18 A L1 1 % (~ % hydrophilic endc.) 8 Phenyl L11 1 (~ % endcapping) 8 C8 L7 1 % (~ % endcapping) 8 C8 A L7 8 % (> % endcapping) 8 C L 7 % (~ % endcapping) 8 HILIC 7 % (no endcapping) 8 NH L8 % (no endcapping) 8 CN L1 7 % (no endcapping) 8 Diol L % (no endcapping) 8 Si L % (no endcapping) 8 Whether your application requires high ph or low ph conditions, Eurospher II C18 H and C18 P columns perform equally well.

3 Exellent batch-to-batch reproducibility Eurospher II 1- C18 H etention time reproducibility (SD) Terbinafine HCl.% Ibuprofen.7% Lovastatin.% Simvastatin.8% Batch 1 Batch Batch Batch Batch Applications The choice of the appropriate column for a particular application can be a daunting task. With a range of bonded phases offering different selectivity, the Eurospher II family includes columns to meet most separation needs. The chart below will help you to choose the best Eurospher II column for a particular application. Phase type non polar polar acidic basic Chelator hydroph. retention shape selectivity extreme aqueous ph > 9 LC-MS C18 C18 H C18 P C18 A Phenyl C8 C8 A C HILIC NH CN Diol Si excellent good suitable not recommended 1... μm Comparable selectivities from µm to µm: The Eurospher and Bluespher column families Bluespher Eurospher family UHPLC HPLC HPLC / MPLC analytical semi-prep. preparative

4 Phenols: Eurospher II C18 P Steroids: Eurospher II C18 P phenol -nitrophenol,-dinitrophenol -chlorophenol -nitrophenol,-dimethylphenol 7 -methyl-,-dinitrolphenol 8 -chloro--methylphenol 9,-dichlorophenol 1,,-trichlorophenol 11 pentachlorophenol i impurity cortisone corticosterone desoxycorticosterone testosterone norgestrel progesterone Column: Eurospher II 1- C18 P 1 x mm Mobile phase: A: water (+.1 % formic acid) B: methanol (+.1 % formic acid) Gradient: min % 9 % B; min hold Flow rate:. ml/min Temperature: C Detection: UV 8 nm Inj. volume: 1 μl Column: Eurospher II 1- C18 P 1 x mm Mobile phase: A: water (+.1 % formic acid) B: methanol (+.1 % formic acid) Gradient: min % 9 % B; min hold Flow rate:. ml/min Temperature: C Detection: UV nm Inj. volume: 1 μl Additives: Eurospher II C18 Water soluble Vitamins: Eurospher II HILIC 1 1 acesulfam saccharin vanillin caffeine aspartame benzoic acid 7 sorbic acid pyridoxal (vitamin B) folic acid (vitamin B9) ascorbic acid (vitamin C) nicotinic acid (vitamin B) thiamine (vitamin B1) cyanocobalamine (vitamin B1) Column: Eurospher II 1- C18 1 x mm Mobile phase: A: mm KH P ; ph B: methanol Gradient: min 1 % % B 8 min % B 8 1 min % % B 1 1 min % B 1 18 min % 1% B Flow rate:. ml/min Temperature: C Detection: UV nm Inj. volume: μl 1-1,, 1, 1,,,,,,, Column: Eurospher II 1- HILIC 1 x mm Mobile phase: A: mm NH -acetate ph B: Acetonitrile Gradient:.7 min % A;.7 1. min % A; 1.. min % A Flow rate: 1. ml/min Temperature: C Detection: UV nm Inj. volume: 1 μl

5 A range of selectivities to fit your application C18 H 1.9 C18H H 11.8 C18P H C18A H Phenyl H C8 H Selectivity plots for phases are based on Tanaka tests: H: hydrophobic retention : hydrophobic selectivity : steric selectivity : hydrogen bonding capacity : ion exchange capacity at ph 7. : ion exchange capacity at ph.7 Modification type Application areas Separation mechanism C18 C18 H C18 P C18 A Phenyl C8 C8 A C HILIC NH CN Diol Si for acidic, basic and neutral analytes in reversed phase mode (sulphonamides; anabolic steroids; anti-psychotics; beta blocker; Sudan dyes; phenols, preservatives etc.) recommended alternative for Kromasil 1 C18 columns; for acidic, basic and neutral analytes in reversed phase mode with extended ph range 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 polar endcapped C18 phase for alternative selectivity; 1 % aqueous applications with very polar compounds (basic pharmaceutical ingredients, water soluble vitamins, catecholamines as well as organic acids) alternative selectivity for aromatic and mode rately polar analytes or mixtures with varying polarity and aromaticity similar selectivity to C18 phase but less retention due to the lower hydrophobicity; useful for determination of water soluble vitamins, steroids, catecholamines, sedatives etc. alternative selectivity to C8 with stronger s for polar compounds; 1 % aqueous applications with very polar compounds universal packing material for different application areas; can also be used in HIC mode (Hydrophobic Interaction Chromatography) especially suited for the separation of hydrophilic, polar and ionic analytes which are only poorly retained on reversed phase columns; behavior is the other way round on Eurospher II HILIC compared to P which makes it an ideal tool to enhance chromatographic separations for these molecules 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 for a wide range of application in normal pase mode as well as reversed phase mode (steroids, carbohydrates polar compounds) alternative to the silica packing with shorter equilibration time and comparable selectivity; due to the lower activity of these packings it can be also used for SEC-applications wide range of different applications, i.e. SEC (size exclusion chromatography) but also for normal phase HPLC; good choice for analytical and preparative purposes to separate polar compounds hydrophobic hydrophobic hydrophobic and steric hydrophobic and polar pi-pi with aromatics reduced hydrophobic comparing to C18 phase hydrophilic and weak electrostatic hydrophobic and hydrophilic hydrophilic and weak electrostatic hydrophilic and ionic hydrophobic and hydrophilic hydrophilic hydrophilic

6 HPLC SMB smometry Column hardware We design and manufacture HPLC column hardware ranging from mm ID to mm ID under strict quality control. A specially treated inner surface ensures consistent column packing and high column stability. A wide range of column lengths from mm up to mm are available. An easily exchangeable inte grated precolumn for analytical columns is available upon request. rdering information E The last 7 digits of the rder No. comprise the stationary phase. E The first digits of the rder No. comprise the column dimensions. Eurospher II packing ecommended for analytical columns ecommended for preparative columns material µm µm 1 µm 1 µm µm C18...E181EG...E181EJ...E181EN...E181EQ...E181EX C18 H...E18EG...E18EJ...E18EN...E18EQ...E18EX C18 P...E18EG...E18EJ...E18EN...E18EQ...E18EX C18 A...E18EG...E18EJ...E18EN...E18EQ...E18EX Phenyl...EEG...EEJ...EEN...EEQ...EEX C8...E81EG...E81EJ...E81EN...E81EQ...E81EX C8 A...E8EG...E8EJ...E8EN...E8EQ...E8EX C...E1EG...E1EJ...E1EN...E1EQ...E1EX HILIC...E1EG...E1EJ...E1EN call call NH...E19EG...E19EJ...E19EN...E19EQ...E19EX CN...EEG...EEJ...EEN...EEQ...EEX Diol...E1EG...E1EJ...E1EN...E1EQ...E1EX Si...EEG...EEJ...EEN...EEQ...EEX For µm columns with comparable selectivities: Analytical and semipreparative columns ID mm mm mm. mm 8 mm 1 mm mm (precolumn) PB... PC... PD... PD... n.a. n.a. mm length B... C... D... E... G... I... mm length B... C... D... E... G... I... 1 mm length 1B... 1C... 1D... 1E... n.a. n.a. 1 mm length 1B... 1C... 1D... 1E... 1G... 1I... 1 mm length 1B... 1C... 1D... 1E... n.a. n.a. mm length B... C... D... E... G... I... mm length n.a. n.a. D... n.a. G... n.a. Preparative columns ID mm mm mm With axial compression mm (precolumn) P mm length 1P... 1Q mm length P... Q All analytical column types from mm ID up to. mm ID are available with integrated precolumns. Kromasil is a brand name of EKA Chemicals AB. Technical data are subject to change without notice. Visit for details on complete HPLC systems, HPLC columns, and osmometers. Wissenschaftliche Gerätebau Dr. Ing. Herbert Knauer GmbH Hegauer Weg 8 11 Berlin, Germany Phone: Telefax: info@knauer.net Internet: Your local distributor: Printed on paper awarded the EU Ecolabel EU Ecolabel : FI/11/1 V77-1US/./.1/