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Open and Toroidal Electrophoresis: Ultra-High Separation Efficiencies in Capillaries, Microchips and Slabs

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Open and Toroidal Electrophoresis: Ultra-High Separation Efficiencies in Capillaries, Microchips and Slabs

Tarso B. Ledur Kist

ISBN: 978-1-119-53940-7 March 2020 212 Pages

Hardcover
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$115.50
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Description

Presents the theory and applications of Toroidal Capillary, Microchip, and Slab Electrophoresis to analytical chemists across a range of disciplines

Written by one of the developers of Toroidal Capillary Electrophoresis (TCE), this book is the first to present this novel analytical technique, in detail, to the field of analytical chemistry.

The exact expressions of separation efficiency, resolution, peak capacity, and many other performance indicators of the open and toroidal layouts are presented and compared.

Featuring numerous illustrations throughout, Open and Toroidal Electrophoresis: Ultra-High Separation Efficiencies in Capillaries, Microchips and Slabs offers chapters covering: Solvents and Buffer Solutions; Fundamentals of Electrophoresis; Open Layout; and Toroidal Layout. Confronting Performance Indicators is next, followed by chapters on High Voltage Modules and Distributors; Heat Removal and Temperature Control; and Detectors. The book finishes with an examination of the applications of Toroidal Electrophoresis.

The first book to offer a detailed account of Toroidal Electrophoresis—written by one of its creators

  • Compares the toroidal layouts with the well-established open layouts of the three most used platforms (Capillary, Microchip, and Slab)
  • Provides solutions to many of the experimental issues arising in electromigration techniques and discusses the voltage distributors and detectors that are compatible with the toroidal layouts
  • Richly illustrated with a large number of useful equations showing the relationships between important operational parameters and the performance indicators 

Open and Toroidal Electrophoresis is aimed at method developers and separation scientists working in clinical analysis, and food analysis, as well as those in pharmacology, disease biomarker applications, and nucleic acid analysis using the Capillary, Microchip, or slab Platform. It will also benefit undergraduate and graduate students of inorganic analytical chemistry, organic analytical chemistry, bioanalysis, pharmaceutical sciences, clinical sciences, and food analysis.

Preface xiii

Acronyms xv

Introduction iii

1 Solvents and Buffer Solutions 1

1.1 Water as a Solvent 1

1.1.1 Temperature and Brownian Motion 1

1.1.2 Electric Permittivity of Water 2

1.1.3 Dissolution 4

1.1.4 Solvation 4

1.1.5 Dissociation 6

1.1.6 Ionization 8

1.1.7 Hydrophilicity, hydrophobicity, and LogP 10

1.1.8 Gibbs Free Energy Change 11

1.1.9 Acid Ionization Constants 12

1.1.10 Concentration-pH and pa-pH Diagrams 15

1.1.11 Henderson-Hasselbalch Equation 18

1.1.12 Buffer Capacity 20

1.2 Binary Mixtures and Other Solvents 23

2 Fundamentals of Electrophoresis 29

2.1 The Platforms 29

2.2 Electrophoresis 31

2.3 Electrophoresis of Single Molecules 36

2.4 Ionic Limiting Mobility 39

2.5 Bands, Frontal Techniques, Peaks, and Zones 40

2.5.1 Bands and Peaks 40

2.5.2 Frontal Techniques 43

2.5.3 Zones 44

2.6 The Isoelectric Point 45

2.6.1 Isoelectric Point of Molecules 45

2.6.2 Isoelectric Point of Nano and Micro-particles 47

2.7 Turbulent and Laminar Flow 48

2.7.1 The Driving Forces of Fluid Flow 48

2.7.2 Turbulence 49

2.7.3 Laminar Flow in Cylindrical Capillaries 50

2.7.4 Laminar Flow in Microchannels 52

2.8 Electroosmosis 55

2.8.1 EOF in Cylindrical Capillaries 55

2.8.2 EOF Volumetric Flow Rate in Rectangular Microchannels 58

2.9 Supression of EOF 58

2.9.1 Protocols for EOF Suppression 59

2.9.2 Advantages of Suppressing EOF with Covalent Coating 61

2.9.3 Measuring Small and Large EOF Velocities 62

2.10 Joule Effect and Heat Dissipation 62

2.11 Temperature Profiles 64

2.12 Molecular Diffusion and Band Broadening 67

2.13 Sample Stacking and Band Compression 72

2.14 Separation Modes 75

2.14.1 Affinity Electrophoresis 75

2.14.2 Electrochromatography 77

2.14.3 End-labeled Free-solution Electrophoresis 78

2.14.4 Free-Solution Electrophoresis 80

2.14.5 Isoelectric Focusing 81

2.14.6 Isotacophoresis 82

2.14.7 Microemulsion Electrokinetic Chromatography 83

2.14.8 Micellar Electrokinetic Chromatography 84

2.14.9 Sieving Electrophoresis 85

2.14.10 Suitable Separation Modes for Each Class of Analytes 87

3 Open Layout 121

3.1 Capillary Electrophoresis 121

3.2 Microchip Electrophoresis 124

3.3 Slab Electrophoresis 127

3.4 Performance Indicators for Open Layouts 130

3.4.1 From Single Bands or Peaks 131

3.4.2 From Two Neighboring Bands or Peaks 134

3.4.3 From n-Bands and n-Peaks 139

4 Toroidal Layout 153

4.1 Toroidal Capillary Electrophoresis 155

4.2 Toroidal Microchip Electrophoresis 157

4.3 Toroidal Slab Electrophoresis 158

4.4 Folding Geometries 158

4.5 Microholes and Connections 160

4.6 Reservoirs 162

4.7 Active and Passive Modes of Operation 163

4.8 Performance Indicators for Toroidal Layouts 166

4.8.1 From Single Bands or Peaks 166

4.8.2 From Two Neighboring Bands or Peaks 168

4.8.3 From n-Bands or n-Peaks 170

5 Confronting Performance Indicators 181

5.1 Performance Indicators from Experimental Data 181

5.2 Performance Indicators Predicted from Operational Parameters 183

6 High Voltage Modules and Distributors 191

6.1 High Voltages in Open Layouts 191

6.2 High Voltages in Toroidal Layouts 193

6.2.1 The Ideal Toroidal Length 193

6.2.2 High Voltage Distribution Made by Four Modules 195

6.2.3 High Voltage Distribution Based on Relays 197

6.2.4 High Voltage Distribution Based on Sliding Switches 197

6.2.5 High Voltage Distribution Based on Rotating Switches 198

7 Heat Removal and Temperature Control 203

7.1 Temperature Gradients are Unavoidable 206

7.2 Temperature has Multiple Effects 207

7.3 Electrical Insulators with High Thermal Conductivity 210

7.4 Cooling Strategies Used in Capillary Electrophoresis 212

7.4.1 Advantages of a Symmetric Cooling Geometry 214

7.5 Cooling Strategies Used in Microchip Electrophoresis 217

7.5.1 Advantages of a Symmetric Cooling Geometry 218

7.6 Cooling Strategies Used in Slab Electrophoresis 218

7.6.1 Advantages of a Rational Cooling Strategy 219

7.7 Shear Rate of the Coolant 219

7.8 Final Considerations 220

8 Detectors 231

8.1 Fixed Point Detectors 232

8.2 Spatial Detectors (Scanners and Cameras) 235

8.3 Derivatization Reactions 235

8.3.1 Fluorogenic Reactions 237

8.3.2 Labeling Reactions 239

8.3.3 Improving Selectivity Through Derivatization 239

9 Applications of Toroidal Electrophoresis 247

A Nomenclature 253

B Species Concentration in Buffer Solutions 261

B.1 Acids (HnA) 262

B.1.1 Monoprotic Acids (n = 1) 262

B.1.2 Diprotic Acids (n = 2) 262

B.1.3 Triprotic Acids (n = 3) 262

B.1.4 Tetraprotic Acids (n = 4) 263

B.2 Bases (B) 264

B.2.1 Monoprotonated Bases (n = 1) 264

B.2.2 Diprotonated Bases (n = 2) 265

B.2.3 Triprotonated Bases (n = 3) 265

B.2.4 Tetraprotonated Bases (n = 4) 266

C Electrophoresis 269

C.1 Free-Solution Electrophoretic Mobility 269

C.1.1 Classical Trajectories 271

C.2 Mobility Dependence on Temperature 273

C.3 Transient Regimes 275

C.3.1 Eletrophoretic Transient Regime (Te) 276

C.3.2 Hardware Transient Regime To 277

D Electroosmosis 281

D.1 Slab and Microhips - Cartesian Coordinates 281

D.2 Capillaries - Cylindrical Coordinates 285

D.3 Zeta Potential 289

E Molecular Diffusion 293

E.1 The Diffusion Equation 293

E.2 The Propagator 295

E.3 Application of Propagators to Bands at Rest 297

E.4 Application of Propagators to Bands in Movement 300

E.5 Bands and Peaks 301

F Poisseulle Counter-flow 303

F.1 Introduction 303

F.2 Velocity Level Contours 304

F.3 Temperature Level Contours 306

F.4 Equalizing vmax and Δve 307

G Cyclic On-column Band Compression 311

G.1 Effect of Cyclic Band Compression Events on Variance 312

G.2 Number of Theoretical Plates 314

G.3 Number of Theoretical Plates per Unit Time 314

G.4 Height Equivalent of a Theoretical Plate 315

G.5 Resolution 315

G.6 Resolution per Unit Time 317

G.7 Band Capacity 318

G.8 Band Capacity per Unit Time 319

G.9 Detailed Calculation of Variance, fn, and hn 321

G.9.1 Peak Variance 321

G.9.2 Inter-Peak Spacing (Δx) 323

G.9.3 Calculation of the Values of hn 324