Flexibility with. Preparative HPLC

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Flexibility with Preparative HPLC Date: August 26 th, 2014 Time: 09.00 10.00 am CEST 1

Who are your Presenters? Dr. Christian Textor Product Manager Preparative HPLC Dipl. Biol. Hagen Schlicke Product Manager Applications & Columns +49 30-809727-303 textor@knauer.net +49 30-809727-675 schlicke@knauer.net 2

What can you expect? Basic and advanced system configurations for AZURA preparative HPLC Interesting news about KNAUER s accessories for preparative HPLC Stacked Injection and Peak Recycling as powerful tools for preparative HPLC Chat with the experts at the end of the session 3

Overview 1. Requirements on preparative HPLC 2. AZURA preparative HPLC systems from KNAUER 3. Useful accessories and conceivable applications 4. Summary 4

Overview 1. Requirements on preparative HPLC 2. AZURA preparative HPLC systems from KNAUER 3. Useful accessories and conceivable applications 4. Summary 5

Application Areas Medical Impurity Analysis Chemistry Purification Natural Products Science Protein / Antibodies 6

Tasks for preparative Chromatography Number of separation problems Repetition of the separation Research and development (mg to g) Technical scale (g to kg) Production scale (kg to t) Time pressure Economic pressure 7

Requirements on preparative HPLC The aim of preparative HPLC is: To isolate the maximum amount of a certain product (recovery) At a desired purity (purity) In a minimum of time (throughput) 8

Requirements on preparative HPLC Optimization strategies: High productivity High purity Low process costs Low instruments investments The individual decision is based on economical, technical, logistical, environmental constraints. 9

Preparative LC Systems from KNAUER SMB Simulated moving bed chromatography AZURA Preparative HPLC Contichrom Preparative liquid chromatography 10

Preparative LC Systems from KNAUER SMB Simulated moving bed chromatography AZURA Preparative HPLC Contichrom Preparative liquid chromatography 11

Overview 1. Requirements on preparative HPLC 2. AZURA preparative HPLC systems from KNAUER 2.1 Instrumentation 2.2 System Configuration 2.3 KNAUER s Vertex Plus AX columns 3. Useful accessories and conceivable applications 4. Summary 12

AZURA Preparative HPLC Designed for the generation of material Suitable to handle large sample volumes 13

AZURA System components P 2.1L Preparative Pump wide flow range (0.01-100 ml/min, 0.1-250 ml/min, 0.1-500 ml/min, 0.1-1000 ml/min) selection of different pump head materials (stainless steel, titanium) integrated compressibility compensation constant pressure operation mode 14

AZURA System components ASM 2.1L Assistant containing feed pump, injection valve and fraction collection valve 15

AZURA System components UVD 2.1L Detector wavelength range from 190 750 nm 16

KNAUER Software solutions OpenLAB EZChrom Edition Full-featured preparative chromatography software Valuable tools for detector controlled fraction collector Exclusive FRC options Peak and solvent recycling Stacked injection 17

KNAUER Software solutions ClarityChrom Preparative Cost effective software solution for automated LC purification tasks Allows peak recognition by level, slope, or level and/or slope 18

Preconfigured systems www.knauer.net/azuraprep survey of preconfigured systems preparative systems for high pressure gradient (HPG), low pressure gradient (LPG) or isocratic applications systems for integration of 1/16, 1/8, 1/4 capillaries 19

Preconfigured systems Flexibility with Preparative HPLC The Assistant ASM 2.1L is the key for flexibility More than 100.000 different configurations possible Find your individual preparative HPLC solution 20

Vertex Plus AX preparative HPLC Columns longer column life time reproducible column effeciency convenient adjustable column bed large variety of column materials refill service 21

How does it work? fixed column part adjustable column part 22

Column Bed Regeneration New column Used column Regenerated column 23

What is available? 5 µm, 10 µm, 15 µm and 20/45 µm 24

Overview 1. Requirements on preparative HPLC 2. AZURA preparative HPLC systems from KNAUER 3. Useful accessories and conceivable applications 4. Summary 25

Useful accessories for preparative HPLC Stacked injection Solvent and Peak recycling AZURA ScaleUp System and ScaleUp Converter Capillaries and Connectors AZURA Mobile Control App 26

Stacked Injection 27

Stacked Injection 28

Benefit (Stacked Injection) Stacked Injection can be used if multiple injections are required. Maximize the productivity of a preparative separation (run time and mobile phase savings) Collect each fraction to a separate vial or collect same fractions in same vials Recommendations: Only for isocratic conditions 29

Solvent and Peak Recycling Eluent Sample Solvent Recycling Peak recycling Assistent Injection valve Fractionation valve Pump Detector Column 30

Solvent and Peak Recycling Efficient tool for improving resolution Fraction collector valves have integrated ports for peak and solvent recycling 31

Benefit (Peak Recycling) Improvement of resolution of two not baseline separated peaks. No eluent consumption during peak recycling Solvent recycling is also supported. Peak recycling allows the usage of shorter columns Reducing of column investments. Recommendations: Only for binary mixtures (e.g. chiral separations) Only isocratic conditions 32

AZURA ScaleUp System Sample pump P 4.1S (ASM 2.1L) Waste Sample injection valve (ASM 2.1L) Sample loop column (preparative) UVD 2.1S (ASM 2.1L) 1/16 OD, 0.5-0.7 mm ID 1/16 OD, 0.25 mm ID column (analytical) Hypersher Smartmix Autosampler S3950 UVD 2.1L Waste P 2.1L Multiposition valve (ASM 2.1L) Column selection valve (ASM 2.1L) P 2.1L 1/16 OD, 1.0 mm ID 26.08.2014 33

AZURA ScaleUp System Additional information available on KNAUER webpage: SYSA007 34

AZURA ScaleUp System Download the ScaleUp Converter software and easily transfer your method from analytical to preparative HPLC: http://www.knauer.net/en/downloads/software.html 35

AZURA ScaleUp System 36

Capillaries and Connectors connectors for: 1/16 1/8 1/4 flow cell for 1/16 1/8 1/4 huge variety of capillaries TRI-Clamp adapter (stainless steel, bio-inert) 37 37

38

Mobile Control How it works How it works 39

Mobile Control Application for Android 4.xx mobile devices Support of 7 and 10 tablets Runs also on mobile phones but UI is not optimized 40

Mobile Control Tablet-based application Wireless communication Direct control of all AZURA devices simultaneously Easy programing of complete HPLC system WIN8 supports enables also the usage where WLAN is prohibited 41

Overview 1. Requirements on preparative HPLC 2. AZURA preparative HPLC systems from KNAUER 3. Useful accessories and conceivable applications 4. Summary 42

Summary Preparative chromatography is a highly applicable method to generate highly pure value products in the food and pharmaceutical industries as well as for the production of fine chemicals. Vertex Plus AX columns convince with longer life times and high reproducibility Stacked Injection is a powerful tool to increase the productivity Closed Loop Peak Recycling is an efficient tool for improving resolution 43

Any questions? Any comments? Use the Chat Function! Dr. Christian Textor Product Manager Preparative HPLC Dipl. Biol. Hagen Schlicke Product Manager Applications & Columns +49 30-809727-303 textor@knauer.net +49 30-809727-675 schlicke@knauer.net 44

When is our next webinar? October 2014 For further information see: www.knauer.net\events.html 45