Comprehensive, up-to-date, and user-centered one-stop reference on the principles and applications of Field-Flow Fractionation (FFF), a highly versatile separation technology

Field-Flow Fractionation: Principles and Applications offers a comprehensive and topical one-stop reference on Field-Flow Fractionation (FFF), an important separation technique which has been proven successful in the analysis of natural and engineered nanoparticles, pharmaceuticals, proteins, polymers, soils, and food.

After introductory chapters on theory, principles, and instrumentation, a tutorial-style user guide addresses typical users' questions and problems. In the application part, the separation of all relevant particle classes is discussed by international experts.

Sample topics covered in Field-Flow Fractionation: Principles and Applications include:

• Historical perspectives of the technique, normal versus steric versus hyperlayer modes, retention resolution, and fractionation power
• FFF techniques, including flow (asymmetric, frit inlet, and outlet), sedimentation, thermal, electric, and other novel techniques such as 2D
• How to determine if FFF is the correct choice and if it is being implemented correctly, along with general optimization strategies including carrier fluid, calibration, and reproducibility
• Sample overloading and recovery, detector selection, troubleshooting, and performance of other fractionation techniques

Both beginners and experienced chemists and researchers can confidently rely on Field-Flow Fractionation: Principles and Applications to confirm their own understanding and to improve their FFF methods and interpretation of their results.

Preface xi

1 Field-Flow Fractionation Techniques 1
P. Stephen Williams

1.1 Introduction 1

1.2 Flow Field-Flow Fractionation 3

1.3 Hollow-Fiber Field-Flow Fractionation 10

1.4 Gravitational Field-Flow Fractionation 14

1.5 Sedimentation Field-Flow Fractionation 16

1.6 Thermal Field-Flow Fractionation 18

1.7 Electrical Field-Flow Fractionation 22

1.8 Magnetic Field-Flow Fractionation 24

1.9 Novel Techniques 27

1.9.1 Combined Fields 27

1.9.2 Two-Dimensional, Continuous Fractionation 28

References 29

2 Field-Flow Fractionation User Guide 45
Haiyang Dou, S. Kim Ratanathanawongs Williams, Mohammed Baalousha, and Céline Guéguen

2.1 Theoretical FFF Background 45

2.2 Different Variants of FlFFF 47

2.3 Sample Properties Measured by FFF 48

2.4 Selection of Detectors 48

2.5 FFF Selection Strategy 50

2.6 Guidance for FFF Separation 50

2.6.1 Sample Pre-Preparation 52

2.6.2 Selection of Carrier Liquid 52

2.6.3 Verify Sample Loss in the FFF Channel 53

2.6.4 Optimize Field Strength 53

2.6.5 Optimize Channel Flow Rate 54

2.6.6 Evaluate Sample Overloading 54

2.6.7 Evaluate Sample Recovery 55

Acknowledgments 55

References 55

3 Polymers and Biohybrids 63
Albena Lederer and Susanne Boye

3.1 Polymer Architectures 64

3.2 FFF Systems for Polymer Characterization 66

3.3 FFF-Coupled Techniques for Polymer Characterization 68

3.4 FFF Separation of Polymers with Different Branching Topology, Composition, and Functionality 69

3.4.1 Hyperbranched Polyesters 70

3.4.2 Dendritic Glycopolymers 72

3.4.3 Single-Chain Nanoparticles 74

3.4.4 Flower-Like Polymer Structures by Self-Assembly 77

3.4.5 Polyethylene Block Copolymers 78

3.5 Biohybrids - Interaction Between Polymers and Biomolecules 80

3.5.1 Polymer-Dye/Drug Complexes 81

3.5.2 Biohybrid Structures Via Polymer-Protein Conjugation 84

3.5.3 Polyplexes: Conjugation of Polymers and DNA 88

3.5.4 Polymersomes 89

References 91

4 Separation Techniques in Support of Elucidating Composition, Structure, and Function Relationships for Complex Polysaccharides 101
Kaitlin C. Lesco, Lieve M. L. Laurens, and S. Kim Ratanathanawongs Williams

4.1 Introduction 101

4.2 Polysaccharide Structures 102

4.3 Analytical Separation Techniques for Polysaccharides 103

4.3.1 Asymmetrical Flow Field-Flow Fractionation 103

4.3.2 Size Exclusion Chromatography 104

4.3.3 Analytical Ultracentrifugation 105

4.3.4 Anion Exchange Chromatography 106

4.4 Online Detection Methods 111

4.4.1 Light Scattering Detectors 112

4.4.2 Detectors for Elucidation of Chemical Properties 113

4.5 Comparing Techniques 114

4.5.1 Resolution 114

4.5.1.1 Size-Based Separation Techniques 114

4.5.1.2 Monosaccharide-Based Separation Techniques 117

4.5.2 Sample Recovery 117

4.5.3 Carrier Fluid Flexibility 118

4.5.4 MW or Size Range 118

4.6 Factors Influencing Separation of Polysaccharides 119

4.6.1 pH 119

4.6.1.1 pH Effects on Polysaccharide Solubility 119

4.6.1.2 pH Effects on Charge 120

4.6.2 Ionic Strength and Composition 120

4.6.2.1 Ionic Strength Effects on Separation 121

4.6.2.2 Ionic Strength Effects on Polysaccharide Structures 121

4.6.2.3 Ionic Composition 122

4.7 Examples of the Separation of Polysaccharides and Their Aggregates 122

4.7.1 Marine Polysaccharides 122

4.7.1.1 Size-Based Separation Techniques 127

4.7.1.2 Compositional and Charge-Based Separation Techniques 127

4.7.2 Plant Polysaccharides 128

4.7.2.1 Size-Based Separation Techniques 129