Preparative Chromatography

Professor Schmidt-Traub was Professor for Plant and Process Design at the Department of Biochemical and Chemical Engineering, University of Dortmund, Germany until his retirement in 2006. He is still active in the research community and his main areas of research focus on preparative chromatography, down stream processing, integrated processes, plant design and innovative energy transfer. Prior to his academic appointment, Prof. Schmidt-Traub gained 15 years of industrial experience in plant engineering. Prof. Seidel-Morgenstern is the Director of the Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany and holds the Chair in Chemical Process Engineering at the Otto-von-Guericke-Universität, Magdeburg, Germany. He received his Ph.D. in 1987 at the Institute of Physical Chemistry of the Academy of Sciences in Berlin. From there he went on to work as postdoctoral fellow at the University of Tennessee, Knoxville, USA. In 1994 he finished his habilitation at the Technical University in Berlin. His research is focused on new reactor concepts, chromatographic reactors, membrane reactors, adsorption and preparative chromatography and separation of enantiomers amongst others. Dr. Michael Schulte is Senior Director Emerging Businesses Energy at Merck KGaA Performance Materials, Darmstadt, Germany. In his Ph.D. thesis at the University of Münster, Germany, he developed new chiral stationary phases for chromatographic enantioseparations. In 1995 he joined Merck and has since then been responsible for research and development in the area of preparative chromatography, including the development of new stationary phases, new separation processes and the implementation of Simulated Moving Bed-technology at Merck. In his current position one of his areas of research is the use of Ionic Liquids for separation processes.

• Preface xvAbout the Editors xviiList of Abbreviations xixNotation xxiii1 Introduction 1Henner Schmidt-Traub and Reinhard Ditz1.1 Chromatography, Development, and Future Trends 11.2 Focus of the Book 41.3 Suggestions on How to Read this Book 4References 62 Fundamentals and General Terminology 9Andreas Seidel-Morgenstern2.1 Principles and Features of Chromatography 92.2 Analysis and Description of Chromatograms 132.2.1 Voidage and Porosity 132.2.2 Retention Times and Capacity Factors 162.2.3 Efficiency of Chromatographic Separations 172.2.4 Resolution 202.2.5 Pressure Drop 232.3 Mass Transfer and Fluid Dynamics 252.3.1 Principles of Mass Transfer 252.3.2 Fluid Distribution in the Column 272.3.3 Packing Nonidealities 282.3.4 Extra-Column Effects 292.4 Equilibrium Thermodynamics 292.4.1 Definition of Isotherms 292.4.2 Models of Isotherms 312.4.2.1 Single-Component Isotherms 312.4.2.2 Multicomponent Isotherms Based on the Langmuir Model 332.4.2.3 Competitive Isotherms Based on the Ideal Adsorbed Solution Theory 342.4.2.4 Steric Mass Action Isotherms 372.4.3 Relation Between Isotherms and Band Shapes 382.5 Column Overloading and Operating Modes 442.5.1 Overloading Strategies 442.5.2 Beyond Isocratic Batch Elution 45References 463 Stationary Phases 49Michael Schulte3.1 Survey of Packings and Stationary Phases 493.2 Inorganic Sorbents 503.2.1 Activated Carbons 503.2.2 Synthetic Zeolites 543.2.3 Porous Oxides: Silica, Activated Alumina, Titania, Zirconia, and Magnesia 543.2.4 Silica 553.2.4.1 Surface Chemistry 573.2.4.2 Mass Loadability 593.2.5 Diatomaceous Earth 593.2.6 Reversed Phase Silicas 603.2.6.1 Silanization of the Silica Surface 603.2.6.2 Silanization 603.2.6.3 Starting Silanes 613.2.6.4 Parent Porous Silica 613.2.6.5 Reaction and Reaction Conditions 623.2.6.6 Endcapping 623.2.6.7 Chromatographic Characterization of Reversed Phase Silicas 633.2.6.8 Chromatographic Performance 633.2.6.9 Hydrophobic Properties Retention Factor (Amount of Organic Solvent for Elution), Selectivity 653.2.6.10 Shape Selectivity 653.2.6.11 Silanol Activity 673.2.6.12 Purity 683.2.6.13 Improved pH Stability Silica 683.2.7 Aluminum Oxide 693.2.8 Titanium Dioxide 703.2.9 Other Oxides 713.2.9.1 Magnesium Oxide 713.2.9.2 Zirconium Dioxide 713.2.10 Porous Glasses 723.3 Cross-Linked Organic Polymers 733.3.1 General Aspects 743.3.2 Hydrophobic Polymer Stationary Phases 773.3.3 Hydrophilic Polymer Stationary Phases 783.3.4 Ion Exchange (IEX) 793.3.4.1 Optimization of Ion-Exchange Resins 813.3.5 Mixed Mode 883.3.6 Hydroxyapatite 883.3.7 Designed Adsorbents 913.3.7.1 Protein A Affinity Sorbents 913.3.7.2 Other IgG Receptor Proteins: Protein G and Protein L 963.3.7.3 Sorbents for Derivatized/Tagged Compounds: Immobilized Metal Affinity Chromatography (IMAC) 963.3.7.4 Other Tag-Based Affinity Sorbents 1013.3.8 Customized Adsorbents 1023.3.8.1 Low Molecular Weight Ligands 1053.3.8.2 Natural Polymers (Proteins, Polynucleotides) 1083.3.8.3 Artificial Polymers 1113.4 Advective Chromatographic Materials 1113.4.1 Adsorptive Membranes and Grafted-Polymer Membranes 1143.4.2 Adsorptive Nonwovens 1153.4.3 Fiber/Particle Composites 1173.4.4 Area-Enhanced Fibers 1173.4.5 Monolith 1183.4.6 Chromatographic Materials for Larger Molecules 1213.5 Chiral Stationary Phases 1213.5.1 Cellulose- and Amylose-Based CSP 1223.5.2 Antibiotic CSP 1283.5.3 Cyclofructan-Based CSP 1283.5.4 Synthetic Polymers 1283.5.5 Targeted Selector Design 1303.5.6 Further Developments 1323.6 Properties of Packings and Their Relevance to Chromatographic Performance 1323.6.1 Chemical and Physical Bulk Properties 1323.6.2 Morphology 1333.6.3 Particulate Adsorbents: Particle Size and Size Distribution 1333.6.4 Pore Texture 1343.6.5 Pore Structural Parameters 1373.6.6 Comparative Rating of Columns 1373.7 Sorbent Maintenance and Regeneration 1383.7.1 Cleaning in Place (CIP) 1383.7.2 CIP for IEX 1403.7.3 CIP of Protein A Sorbents 1403.7.4 Conditioning of Silica Surfaces 1433.7.5 Sanitization in Place (SIP) 1453.7.6 Column and Adsorbent Storage 145References 1464 Selection of Chromatographic Systems 159Michael Schulte4.1 Definition of the Task 1644.2 Mobile Phases for Liquid Chromatography 1674.2.1 Stability 1684.2.2 Safety Concerns 1724.2.3 Operating Conditions 1724.2.4 Aqueous Buffer Systems 1764.3 Adsorbent and Phase Systems 1784.3.1 Choice of Phase System Dependent on Solubility 1784.3.2 Improving Loadability for Poor Solubilities 1804.3.3 Dependency of Solubility on Sample Purity 1834.3.4 Generic Gradients for Fast Separations 1844.4 Criteria for Choosing Normal Phase Systems 1844.4.1 Retention in NP Systems 1864.4.2 Solvent Strength in Liquid–Solid Chromatography 1884.4.3 Pilot Technique Thin-Layer Chromatography Using the PRISMA Model 1904.4.3.1 Step (1): Solvent Strength Adjustment 1994.4.3.2 Step (2): Optimization of Selectivity 1994.4.3.3 Step (3): Final Optimization of the Solvent Strength 2004.4.3.4 Step (4): Determination of the Optimum Mobile Phase Composition 2004.4.4 Strategy for an Industrial Preparative Chromatography Laboratory 2024.4.4.1 Standard Gradient Elution Method on Silica 2034.4.4.2 Simplified Procedure 2044.5 Criteria for Choosing Reversed Phase Systems 2064.5.1 Retention and Selectivity in RP Systems 2084.5.2 Gradient Elution for Small Amounts of Product on RP Columns 2124.5.3 Rigorous Optimization for Isocratic Runs 2134.5.4 Rigorous Optimization for Gradient Runs 2174.5.5 Practical Recommendations 2224.6 Criteria for Choosing CSP Systems 2234.6.1 Suitability of Preparative CSP 2234.6.2 Development of Enantioselectivity 2244.6.3 Optimization of Separation Conditions 2264.6.3.1 Determination of Racemate Solubility 2264.6.3.2 Selection of Elution Order 2264.6.3.3 Optimization of Mobile/Stationary Phase Composition, Including Temperature 2264.6.3.4 Determination of Optimum Separation Step 2274.6.4 Practical Recommendations 2274.7 Downstream Processing of mAbs Using Protein A and IEX 2314.8 Size-Exclusion Chromatography (SEC) 2364.9 Overall Chromatographic System Optimization 2374.9.1 Conflicts During Optimization of Chromatographic Systems 2374.9.2 Stationary Phase Gradients 241References 2465 Process Concepts 251Malte Kaspereit and Henner Schmidt-Traub5.1 Discontinuous Processes 2525.1.1 Isocratic Operation 2525.1.2 Gradient Chromatography 2535.1.3 Closed-Loop Recycling Chromatography 2565.1.4 Steady-State Recycling Chromatography (SSRC) 2585.1.5 Flip-Flop Chromatography 2595.1.6 Chromatographic Batch Reactors 2605.2 Continuous Processes 2615.2.1 Column Switching Chromatography 2625.2.2 Annular Chromatography 2625.2.3 Multiport Switching Valve Chromatography (ISEP/CSEP) 2635.2.4 Isocratic Simulated Moving Bed (SMB) Chromatography 2645.2.5 SMB Chromatography with Variable Process Conditions 2685.2.5.1 Varicol 2695.2.5.2 PowerFeed 2705.2.5.3 Partial-Feed, Partial-Discard, and Fractionation-Feedback Concepts 2715.2.5.4 Improved/Intermittent SMB (iSMB) 2715.2.5.5 Modicon 2735.2.5.6 FF-SMB 2735.2.6 Gradient SMB Chromatography 2745.2.7 Supercritical Fluid Chromatography (SFC) 2755.2.7.1 Supercritical Batch Chromatography 2765.2.7.2 Supercritical SMB processes 2775.2.8 Multicomponent Separations 2775.2.9 Multicolumn Systems for Bioseparations 2785.2.9.1 Multicolumn Capture Chromatography (MCC) 2795.2.9.2 Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) 2865.2.10 Countercurrent Chromatographic Reactors 2885.2.10.1 SMB Reactor 2885.2.10.2 SMB Reactors with Distributed Functionalities 2905.3 Choice of Process Concepts 2925.3.1 Scale 2925.3.2 Range of k’ 2925.3.3 Number of Fractions 2935.3.4 Example 1: Lab Scale; Two Fractions 2935.3.5 Example 2: Lab Scale; Three or More Fractions 2945.3.6 Example 3: Production Scale; Wide Range of k’ 2965.3.7 Example 4: Production Scale; Two Main Fractions 2975.3.8 Example 5: Production Scale; Three Fractions 2985.3.9 Example 6: Production Scale; Multistage Process 300References 3026 Modeling of Chromatographic Processes 311Andreas Seidel-Morgenstern6.1 Introduction 3116.2 Models for Single Chromatographic Columns 3116.2.1 Equilibrium Stage Models 3126.2.1.1 Discontinuous Model According to Craig 3136.2.1.2 Continuous Model According to Martin and Synge 3156.2.2 Derivation of Continuous Mass Balance Equations 3166.2.2.1 Mass Balance Equations 3186.2.2.2 Convective Transport 3206.2.2.3 Axial Dispersion 3206.2.2.4 Intraparticle Diffusion 3216.2.2.5 Mass Transfer Between Phases 3216.2.2.6 Finite Rates of Adsorption and Desorption 3226.2.2.7 Adsorption Equilibria 3236.2.3 Equilibrium Model of Chromatography 3236.2.4 Models with One Band Broadening Effect 3296.2.4.1 Equilibrium Dispersion Model 3296.2.4.2 Finite Adsorption Rate Model 3316.2.5 Continuous Lumped Rate Models 3316.2.5.1 Transport Dispersion Models 3326.2.5.2 Lumped Finite Adsorption Rate Model 3336.2.6 General Rate Models 3336.2.7 Initial and Boundary Conditions of the Column 3356.2.8 Dimensionless Model Equations 3366.2.9 Comparison of Different Model Approaches 3386.3 Including Effects Outside the Columns 3436.3.1 Experimental Setup and Simulation Flow Sheet 3436.3.2 Modeling Extra-Column Equipment 3456.3.2.1 Injection System 3456.3.2.2 Piping 3456.3.2.3 Detector 3456.4 Calculation Methods and Software 3466.4.1 Analytical Solutions 3466.4.2 Numerical Solution Methods 3466.4.2.1 Discretization 3466.4.2.2 General Solution Procedure and Software 349References 3507 Determination of Model Parameters 355Andreas Seidel-Morgenstern, Andreas Jupke, and Henner Schmidt-Traub7.1 Parameter Classes for Chromatographic Separations 3557.1.1 Design Parameters 3557.1.2 Operating Parameters 3567.1.3 Model Parameters 3567.2 Concept to Determine Model Parameters 3577.3 Detectors and Parameter Estimation 3597.3.1 Calibration of Detectors 3597.3.2 Parameter Estimation 3607.3.3 Evaluation of Chromatograms 3627.4 Determination of Packing Parameters 3637.4.1 Void Fraction and Porosity of the Packing 3637.4.2 Axial Dispersion 3637.4.3 Pressure Drop 3647.5 Adsorption Isotherms 3657.5.1 Determination of Adsorption Isotherms 3657.5.2 Estimation of Henry Coefficients 3657.5.3 Static Isotherm Determination Methods 3707.5.3.1 Batch Method 3707.5.3.2 Adsorption–Desorption Method 3707.5.3.3 Circulation Method 3717.5.4 Dynamic Methods 3717.5.5 Frontal Analysis 3717.5.6 Analysis of Dispersed Fronts 3787.5.7 Peak Maximum Method 3807.5.8 Minor Disturbance/Perturbation Method 3807.5.9 Curve Fitting of the Chromatogram 3837.5.10 Data Analysis and Accuracy 3847.6 Mass Transfer Kinetics 3867.6.1 Correlations 3867.6.2 Application of Method of Moments 3887.7 Plant Parameters 3897.8 Experimental Validation of Column Models and Model Parameters 3917.8.1 Batch Chromatography 3917.8.2 Simulated Moving Bed Chromatography 3947.8.2.1 Model Formulation and Parameters 3947.8.2.2 Experimental Validation 400References 4048 Process Design and Optimization 409Andreas Jupke, Andreas Biselli, Malte Kaspereit,Martin Leipnitz, and Henner Schmidt-Traub8.1 Basic Principles and Definitions 4098.1.1 Performance, Costs, and Objective Functions 4098.1.1.1 Performance Criteria 4108.1.1.2 Economic Criteria 4118.1.1.3 Objective Functions 4128.1.2 Degrees of Freedom 4138.1.2.1 Categories of Parameters 4138.1.2.2 Dimensionless Operating and Design Parameters 4148.1.3 Scaling by Dimensionless Parameters 4188.1.3.1 Influence of Different HETP Coefficients for Every Component 4198.1.3.2 Influence of Feed Concentration 4208.1.3.3 Examples for a Single-Column Batch Chromatography 4218.1.3.4 Examples for SMB Processes 4248.2 Batch Chromatography 4268.2.1 Fractionation Mode (Cut Strategy) 4268.2.2 Design and Optimization of Batch Chromatographic Columns 4278.2.2.1 Process Performance Depending on Number of Stages and Loading Factor 4278.2.2.2 Design and Optimization Strategy 4328.2.2.3 Other Strategies 4368.3 Recycling Chromatography 4378.3.1 Design of Steady-State Recycling Chromatography 4378.3.2 Scale-Up of Closed-Loop Recycling Chromatography 4408.4 Conventional Isocratic SMB Chromatography 4458.4.1 Considerations to Optimal Concentration Profiles in SMB Process 4458.4.2 Process Design Based on TMB Models (Shortcut Methods) 4468.4.2.1 Triangle Theory for an Ideal Model with Linear Isotherms 4478.4.2.2 Triangle Theory for an Ideal Model with Nonlinear Isotherms 4498.4.2.3 Shortcut to Apply the Triangle Theory on a System with Unknown Isotherms Assuming Langmuir Character 4528.4.3 Process Design and Optimization Based on Rigorous SMB Models 4558.4.3.1 Estimation of Operating Parameter 4568.4.3.2 Optimization of Operating Parameters for Linear Isotherms Based on Process Understanding 4578.4.3.3 Optimization of Operating Parameters for Nonlinear Isotherms Based on Process Understanding 4588.4.3.4 Optimization of Design Parameters 4608.5 Isocratic SMB Chromatography Under Variable Operating Conditions 4658.5.1 Performance Comparison of Varicol and Conventional SMB 4668.5.2 Performance Comparison of Varicol, PowerFeed, and Modicon with Conventional SMB 4708.5.3 Performance Trends Applying SMB Concepts Under Variable Operating Conditions 4758.6 Gradient SMB Chromatography 4768.6.1 Step Gradient 4768.6.2 Multicolumn Solvent Gradient Purification Process 4828.7 Multicolumn Systems for Bioseparations 4878.7.1 Design of Twin-Column Capture SMB 4888.7.2 Modeling of Multicolumn Capture processes 490References 4939 Process Control 503Sebastian Engell and Achim Kienle9.1 Standard Process Control 5049.2 Advanced Process Control 5049.2.1 Online Optimization of Batch Chromatography 5059.2.2 Advanced Control of SMB Chromatography 5079.2.2.1 Purity Control for SMB Processes 5089.2.2.2 Direct Optimizing Control of SMB Processes 5109.2.3 Advanced Parameter and State Estimation for SMB Processes 5159.2.4 Adaptive Cycle-to-Cycle Control 5179.2.5 Control of Coupled Simulated Moving Bed Processes for the Production of Pure Enantiomers 519References 52110 Chromatography Equipment: Engineering and Operation 525Henner Schmidt-Traub and Arthur Susanto10.1 Challenges for Conceptual Process Design 52510.1.1 Main Cost Factors for a Chromatographic System 52710.1.2 Conceptual Process Design 52810.1.2.1 A Case Study: Large-Scale Biotechnology Project 52910.2 Engineering Challenges 53310.2.1 Challenges Regarding Sanitary Design 53510.2.2 Challenges During Acceptance Tests and Qualifications 53910.3 Commercial Chromatography Columns 54010.3.1 General Design 54110.3.1.1 Manually Moved Piston 54210.3.1.2 Electrically or Hydraulically Moved Piston 54210.3.2 High- and Low-Pressure Columns 54310.3.2.1 Chemical Compatibility 54410.3.2.2 Frit Design 54610.3.2.3 Special Aspects of Bioseparation 54910.4 Commercial Chromatographic Systems 55110.4.1 General Design Aspects: High-Pressure and Low-Pressure Systems 55110.4.2 Material 55310.4.3 Batch Low-Pressure Liquid Chromatographic (LPLC) Systems 55310.4.3.1 Inlets 55310.4.3.2 Valves to Control Flow Direction 55510.4.3.3 Pumps 55610.4.3.4 Pump- and Valve-Based and Gradient Formation 55610.4.4 Batch High-Pressure Liquid Chromatography 55810.4.4.1 General Layout 55810.4.4.2 Inlets and Outlets 55910.4.4.3 Pumps 55910.4.4.4 Valves and Pipes 56210.4.5 Continuous Systems: Simulated Moving Bed 56310.4.5.1 General Layout 56310.4.5.2 A Key Choice: The Recycling Strategy 56510.4.5.3 Pumps, Inlets, and Outlets 56610.4.5.4 Valves and Piping 56610.4.6 Auxiliary Systems 56710.4.6.1 Slurry Preparation Tank 56710.4.6.2 Slurry Pumps and Packing Stations 56810.4.6.3 Cranes and Transport Units 56810.4.6.4 Filter Integrity Test 56810.4.7 Detectors 56910.5 Packing Methods 57110.5.1 Column and Packing Methodology Selection 57110.5.2 Slurry Preparation 57210.5.3 Column Preparation 57410.5.4 Flow Packing 57510.5.5 Dynamic Axial Compression (DAC) Packing 57710.5.6 Stall Packing 57710.5.7 Combined Method (Stall+DAC) 57810.5.8 Vacuum Packing 58010.5.9 Vibration Packing 58110.5.10 Column Equilibration 58210.5.11 Column Testing and Storage 58310.5.11.1 Test Systems 58310.5.11.2 Hydrodynamic Properties and Column Efficiency 58410.5.11.3 Column and Adsorbent Storage 58510.6 Process Troubleshooting 58510.6.1 Technical Failures 58610.6.2 Loss of Performance 58710.6.2.1 Pressure Increase 58710.6.2.2 Loss of Column Efficiency 59010.6.2.3 Variation of Elution Profile 59110.6.2.4 Loss of Purity/Yield 59210.6.3 Column Stability 59210.7 Disposable Technology for Bioseparations 59310.7.1 Prepacked Columns 59610.7.2 Membrane Chromatography 597References 599Appendix A Data of Test Systems 601A.1 EMD53986 601A.2 Tröger’s Base 602A.3 Glucose and Fructose 604A.4 β-Phenethyl Acetate 606References 607Index 609

"I would not hesitate to recommend it to anyone working in this field."Chromatographia "Overall the coverage is a bit uneven - nevertheless the volume does compile some useful material... In conclusion, this is a comprehensive reference text, which should find its way into the libraries of all companies who are serious about process scale preparative chromatography, whether internally or via outsource contracts."Organic Process Research and Development "This special volume is essential for chemists and engineers working in chemical and pharmaceutical industries, as well as for food technologies, due to the interdisciplinary nature of these preparative chromatographic processes."Advances in Food Sciences