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Explores the benefits and limitations of the latest capillary electrophoresis techniques Capillary electrophoresis and microchip capillary electrophoresis are powerful analytical tools that are particularly suited for separating and analyzing biomolecules. In comparison with traditional analytical techniques, capillary electrophoresis and microchip capillary electrophoresis offer the benefits of speed, small sample and solvent consumption, low cost, and the possibility of miniaturization. With contributions from a team of leading analytical scientists, Capillary Electrophoresis and Microchip…mehr
Explores the benefits and limitations of the latest capillary electrophoresis techniques Capillary electrophoresis and microchip capillary electrophoresis are powerful analytical tools that are particularly suited for separating and analyzing biomolecules. In comparison with traditional analytical techniques, capillary electrophoresis and microchip capillary electrophoresis offer the benefits of speed, small sample and solvent consumption, low cost, and the possibility of miniaturization. With contributions from a team of leading analytical scientists, Capillary Electrophoresis and Microchip Capillary Electrophoresis explains how researchers can take full advantage of all the latest techniques, emphasizing applications in which capillary electrophoresis has proven superiority over other analytical approaches. The authors not only explore the benefits of each technique, but also the limitations, enabling readers to choose the most appropriate technique to analyze a particular sample. The book's twenty-one chapters explore fundamental aspects of electrophoretically driven separations, instrumentation, sampling techniques, separation modes, detection systems, optimization strategies for method development, and applications. Specific topics include: * Critical evaluation of the use of surfactants in capillary electrophoresis * Sampling and quantitative analysis in capillary electrophoresis * Capillary electrophoresis with electrochemical detection * Overcoming challenges in using microchip electrophoresis for extended monitoring applications * Capillary electrophoresis of intact unfractionated heparin and related impurities * Microchip capillary electrophoresis for in situ planetary exploration Each chapter begins with an introduction and ends with conclusions as well as references to the primary literature. Novices to the field will find this book an easy-to-follow introduction to core capillary electrophoresis techniques and methods. More experienced investigators can turn to the book for troubleshooting tips and expert advice to guide them through the most advanced applications.
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Carlos D. García, PhD, is an Associate Professor of Analytical Chemistry at the University of Texas at San Antonio, USA. His group is currently focused on the development of novel bioanalytical strategies involving microfluidics and nanomaterials. Karin Y. Chumbimuni-Torres, PhD, is a Research Associate at the University of Texas at San Antonio, USA. She is interested in pursuing the development of electrochemical biosensors and their integration to microchip-based platforms. Emanuel Carrilho, PhD, is an Associate Professor at the University of Säo Paulo, Brazil. With more than twenty-five years of experience in separation science, his group is focused on the development of analytical methods and instrumentation for bioanalyses.
Inhaltsangabe
PREFACE xvii
ACKNOWLEDGMENTS xix
CONTRIBUTORS xxi
1 Critical Evaluation of the Use of Surfactants in Capillary Electrophoresis 1 Jessica L. Felhofer, Karin Y. Chumbimuni-Torres, Maria F. Mora, Gabrielle G. Haby, and Carlos D. Garc?´a
1.1 Introduction 1
1.2 Surfactants for Wall Coatings 4
1.3 Surfactants as Buffer Additives 6
1.4 Surfactants for Analyte Preconcentration 9
1.5 Surfactants and Detection in CE 14
1.6 Conclusions 16
References 17
2 Sample Stacking: A Versatile Approach for Analyte Enrichment in CE and Microchip-CE 23 Bruno Perlatti, Emanuel Carrilho, and Fernando Armani Aguiar
2.1 Introduction 23
2.2 Isotachophoresis 24
2.3 Chromatography-Based Sample Stacking 25
2.4 Methods Based on Electrophoretic Mobility and Velocity Manipulation (Electrophoretic Methods) 26
2.5 Sample Stacking in Pseudo-Stationary Phases 29
2.6 Stacking Techniques in Microchips 33
2.7 Concluding Remarks 36
References 37
3 Sampling and Quantitative Analysis in Capillary Electrophoresis 41 Petr Kuba´9n, Andrus Seiman, and Mihkel Kaljurand
3.1 Introduction 41
3.2 Injection Techniques in CE 42
3.3 Micromachined/Microchip Injection Devices 53
3.4 Automated Flow Sample Injection and Hyphenated Systems 55
3.5 Computerized Sampling and Data Analysis 57
3.6 Sampling in Portable CE Instrumentation 58
3.7 Quantitative Analysis in CE 59
3.8 Conclusions 62
References 62
4 Practical Considerations for the Design and Implementation of High-Voltage Power Supplies for Capillary and Microchip Capillary Electrophoresis 67 Lucas Blanes, Wendell Karlos Tomazelli Coltro, Renata Mayumi Saito, Claudimir Lucio do Lago, Claude Roux, and Philip Doble
4.1 Introduction 67
4.2 High-Voltage Measurement 73
4.3 Concluding Remarks 74
References 74
5 Artificial Neural Networks in Capillary Electrophoresis 77 Josef Havel, Eladia Mar?a Pe~na-Mendez, and Alberto Rojas-Hernandez
5.1 Introduction 77
5.2 Optimization in CE: From Single Variable Approach Toward Artificial Neural Networks 77
5.3 Artificial Neural Networks in Electromigration Methods 81
5.4 Conclusions 90
Acknowledgments 91
References 91
6 Improving the Separation in Microchip Electrophoresis by Surface Modification 95 M. Teresa Fernandez-Abedul, Isabel Alvarez-Martos, Francisco Javier Garc?a Alonso, and Agust?n Costa-Garc?a
6.1 Introduction 95
6.2 Strategies for Improving Separation 96
6.3 Chemical Modifiers 102
6.4 Conclusions 119
Acknowledgments 120
References 120
7 Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor: Dissociation Kinetic Analysis Method for "Complexes" Using Capillary Electrophoretic Separation Process 127 Toru Takahashi and Nobuhiko Iki
7.1 Introduction 127
7.2 Basic Concept of CER 128
7.3 Dissociation Kinetic Analysis of Metal Complexes Using a CER 129
7.4 Expanding the Scope of the CER to Measurements of Fast Dissociation Kinetics with a Half-Life from Seconds to Dozens of Seconds: Dissociation Kinetic Analysis of Metal Complexes Using a Microchip Capillary Electrophoretic Reactor (mCER) 133
7.5 Expanding the Scope of the CER to the Measurement of Slow Dissociation Kinetics with a Half-Life of Hours 135
7.6 Expanding the Scope of CER to Measurement of the Dissociation Kinetics of Biomolecular Complexes 139
7.7 Conclusions 142
References 142
8 Capacitively Coupled Contactless Conductivity Detection (C4D) Applied to Capillary Electrophoresis (CE) and Microchip Electrophoresis (MCE) 145 Jose Alberto Fracassi da Silva, Claudimir Lucio do Lago, Dosil Pereira de Jesus, and Wendell Karlos Tomazelli Coltro
8.1 Introduction 145
8.2 Theory of C4D 145
8.3 C4D Applied to Capillary Electrophoresis 148
8.4 C4D Applied to Microchip Capillary Electrophoresis 151
8.5 Concluding Remarks 156
Acknowledgments 157
References 157
9 Capillary Electrophoresis with Electrochemical Detection 161 Blanaid White
9.1 Principles of Electrochemical Detection 161
9.2 Interfacing Amperometric Detection to Capillary Electrophoresis 163
9.3 Interfacing Electrochemical Detection to Microfluidic Capillary Electrophoresis 168
References 173
10 Overcoming Challenges in Using Microchip Electrophoresis for Extended Monitoring Applications 177 Scott D. Noblitt and Charles S. Henry
10.1 Introduction 177
10.2 Background Electrolyte (BGE) Longevity 179
10.3 Achieving Rapid Sequential Injections 186
10.4 Robust Quantitation 192
10.5 Conclusions 197
References 198
11 Distinction of Coexisting Protein Conformations by Capillary Electrophoresis 201 Hanno Stutz
11.1 Introduction 201
11.2 Protein Misfolding and Induction of Unfolding 203
11.3 Conformational Pathologies 204
11.4 Distinction Between Conformations 205
11.5 Relevance of Conformations for Biotechnological Products 206
11.6 Conformational Elucidation--An Overview of Alternative Methods to CE 206
11.7 HPLC in Conformational Distinction 207
11.8 Capillary Electrophoresis (CE) in Conformational Separations 209
11.9 Comparison Between CE and HPLC 223
11.10 Conclusive Discussion and Method Evaluation 223
References 225
12 Capillary Electromigration Techniques for the Analysis of Drugs and Metabolites in Biological Matrices: A Critical Appraisal 229 Cristiane Masetto de Gaitani, Anderson Rodrigo Moraes de Oliveira, and Pierina Sueli Bonato
12.1 Introduction 229
12.2 Strategies to Obtain Reliable Capillary Electromigration Methods for the Bioanalysis of Drugs and Metabolites 230
12.3 Selected Applications of Capillary Electromigration Techniques in Bioanalysis 238
12.4 Concluding Remarks 243
References 243
13 Capillary Electrophoresis and Multicolor Fluorescent DNA Analysis in an Optofluidic Chip 247 Chaitanya Dongre, Hugo J.W.M. Hoekstra, and Markus Pollnau
13.1 Introduction 247
13.2 Optofluidic Integration in an Electrophoretic Microchip 248
13.3 Fluorescence Monitoring of On-Chip DNA Separation 249
14 Capillary Electrophoresis of Intact Unfractionated Heparin and Related Impurities 267 Robert Weinberger
14.1 Introduction 267
14.2 Capillary Electrophoresis and Heparin 269
14.3 Method Development in Capillary Electrophoresis 269
14.4 Common Impurities Found in Heparin 272
14.5 The United States Pharmacoepia and CE of Heparin 273
14.6 Interlaboratory Collaborative Study 274
14.7 Conclusions 275
References 275
15 Microchip Capillary Electrophoresis for In Situ Planetary Exploration 277 Peter A. Willis and Amanda M. Stockton
15.1 Introduction 277
15.2 Instrument Design 279
15.3 Instrumentation External to the Microdevice 280
15.4 Microdevice Basics 282
15.5 Microdevices and their Applications 285
15.6 Conclusions 289
Acknowledgments 290
References 290
16 Rapid Analysis of Charge Heterogeneity of Monoclonal Antibodies by Capillary Zone Electrophoresis and Imaged Capillary Isoelectric Focusing 293 Yan He, Jim Mo, Xiaoping He, and Margaret Ruesch
16.1 Introduction 293
16.2 Capillary Zone Electrophoresis 295
16.3 Imaged Capillary Isoelectric Focusing 299
16.4 Summary 306
References 307
17 Application of Capillary Electrophoresis for High-Throughput Screening of Drug Metabolism 309 Roman 9Rem?´nek, Jochen Pauwels, Xu Wang, Jos Hoogmartens, Zden9ek Glatz, and Ann Van Schepdael
17.1 Introduction 309
17.2 Sample Deproteinization 310
17.3 On-line Preconcentration 311
17.4 Method Development 312
17.5 Method Validation 314
17.6 Method Applications 315
17.7 Conclusions 316
Acknowledgments 317
References 317
18 Electrokinetic Transport of Microparticles in the Microfluidic Enclosure Domain 319 Qian Liang, Chun Yang, and Jianmin Miao
18.1 Introduction 319
18.2 Numerical Model 320
18.3 Numerical Simulation 322
18.4 Results and Discussion 322
18.5 Model Application 325
18.6 Conclusions 326
References 326
19 Integration of Nanomaterials in Capillary and Microchip Electrophoresis as a Flexible Tool 327 Germa´n A. Messina, Roberto A. Olsina, and Patricia W. Stege
19.1 Introduction 327
19.2 Nanomaterials in Analytical Chemistry 332
19.3 Nanoparticles in Capillary Electrophoresis 333
19.4 Conclusions 352
References 353
20 Microchip Capillary Electrophoresis to Study the Binding of Ligands to Teicoplanin Derivatized on Magnetic Beads 359 Toni Ann Riveros, Roger Lo, Xiaojun Liu, Marisol Salgado, Hector Carmona, and Frank A. Gomez
20.1 Introduction 359
20.2 Experimental Section 359
20.3 Results and Discussion 361
20.4 Conclusions 364
Acknowledgments 365
References 365
21 Glycomic Profiling Through Capillary Electrophoresis and Microchip Capillary Electrophoresis 367 Yehia Mechref
21.1 Introduction 367
21.2 General Considerations of Capillary Electrophoresis and Microchip Capillary Electrophoresis of Glycans 369
1 Critical Evaluation of the Use of Surfactants in Capillary Electrophoresis 1 Jessica L. Felhofer, Karin Y. Chumbimuni-Torres, Maria F. Mora, Gabrielle G. Haby, and Carlos D. Garc?´a
1.1 Introduction 1
1.2 Surfactants for Wall Coatings 4
1.3 Surfactants as Buffer Additives 6
1.4 Surfactants for Analyte Preconcentration 9
1.5 Surfactants and Detection in CE 14
1.6 Conclusions 16
References 17
2 Sample Stacking: A Versatile Approach for Analyte Enrichment in CE and Microchip-CE 23 Bruno Perlatti, Emanuel Carrilho, and Fernando Armani Aguiar
2.1 Introduction 23
2.2 Isotachophoresis 24
2.3 Chromatography-Based Sample Stacking 25
2.4 Methods Based on Electrophoretic Mobility and Velocity Manipulation (Electrophoretic Methods) 26
2.5 Sample Stacking in Pseudo-Stationary Phases 29
2.6 Stacking Techniques in Microchips 33
2.7 Concluding Remarks 36
References 37
3 Sampling and Quantitative Analysis in Capillary Electrophoresis 41 Petr Kuba´9n, Andrus Seiman, and Mihkel Kaljurand
3.1 Introduction 41
3.2 Injection Techniques in CE 42
3.3 Micromachined/Microchip Injection Devices 53
3.4 Automated Flow Sample Injection and Hyphenated Systems 55
3.5 Computerized Sampling and Data Analysis 57
3.6 Sampling in Portable CE Instrumentation 58
3.7 Quantitative Analysis in CE 59
3.8 Conclusions 62
References 62
4 Practical Considerations for the Design and Implementation of High-Voltage Power Supplies for Capillary and Microchip Capillary Electrophoresis 67 Lucas Blanes, Wendell Karlos Tomazelli Coltro, Renata Mayumi Saito, Claudimir Lucio do Lago, Claude Roux, and Philip Doble
4.1 Introduction 67
4.2 High-Voltage Measurement 73
4.3 Concluding Remarks 74
References 74
5 Artificial Neural Networks in Capillary Electrophoresis 77 Josef Havel, Eladia Mar?a Pe~na-Mendez, and Alberto Rojas-Hernandez
5.1 Introduction 77
5.2 Optimization in CE: From Single Variable Approach Toward Artificial Neural Networks 77
5.3 Artificial Neural Networks in Electromigration Methods 81
5.4 Conclusions 90
Acknowledgments 91
References 91
6 Improving the Separation in Microchip Electrophoresis by Surface Modification 95 M. Teresa Fernandez-Abedul, Isabel Alvarez-Martos, Francisco Javier Garc?a Alonso, and Agust?n Costa-Garc?a
6.1 Introduction 95
6.2 Strategies for Improving Separation 96
6.3 Chemical Modifiers 102
6.4 Conclusions 119
Acknowledgments 120
References 120
7 Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor: Dissociation Kinetic Analysis Method for "Complexes" Using Capillary Electrophoretic Separation Process 127 Toru Takahashi and Nobuhiko Iki
7.1 Introduction 127
7.2 Basic Concept of CER 128
7.3 Dissociation Kinetic Analysis of Metal Complexes Using a CER 129
7.4 Expanding the Scope of the CER to Measurements of Fast Dissociation Kinetics with a Half-Life from Seconds to Dozens of Seconds: Dissociation Kinetic Analysis of Metal Complexes Using a Microchip Capillary Electrophoretic Reactor (mCER) 133
7.5 Expanding the Scope of the CER to the Measurement of Slow Dissociation Kinetics with a Half-Life of Hours 135
7.6 Expanding the Scope of CER to Measurement of the Dissociation Kinetics of Biomolecular Complexes 139
7.7 Conclusions 142
References 142
8 Capacitively Coupled Contactless Conductivity Detection (C4D) Applied to Capillary Electrophoresis (CE) and Microchip Electrophoresis (MCE) 145 Jose Alberto Fracassi da Silva, Claudimir Lucio do Lago, Dosil Pereira de Jesus, and Wendell Karlos Tomazelli Coltro
8.1 Introduction 145
8.2 Theory of C4D 145
8.3 C4D Applied to Capillary Electrophoresis 148
8.4 C4D Applied to Microchip Capillary Electrophoresis 151
8.5 Concluding Remarks 156
Acknowledgments 157
References 157
9 Capillary Electrophoresis with Electrochemical Detection 161 Blanaid White
9.1 Principles of Electrochemical Detection 161
9.2 Interfacing Amperometric Detection to Capillary Electrophoresis 163
9.3 Interfacing Electrochemical Detection to Microfluidic Capillary Electrophoresis 168
References 173
10 Overcoming Challenges in Using Microchip Electrophoresis for Extended Monitoring Applications 177 Scott D. Noblitt and Charles S. Henry
10.1 Introduction 177
10.2 Background Electrolyte (BGE) Longevity 179
10.3 Achieving Rapid Sequential Injections 186
10.4 Robust Quantitation 192
10.5 Conclusions 197
References 198
11 Distinction of Coexisting Protein Conformations by Capillary Electrophoresis 201 Hanno Stutz
11.1 Introduction 201
11.2 Protein Misfolding and Induction of Unfolding 203
11.3 Conformational Pathologies 204
11.4 Distinction Between Conformations 205
11.5 Relevance of Conformations for Biotechnological Products 206
11.6 Conformational Elucidation--An Overview of Alternative Methods to CE 206
11.7 HPLC in Conformational Distinction 207
11.8 Capillary Electrophoresis (CE) in Conformational Separations 209
11.9 Comparison Between CE and HPLC 223
11.10 Conclusive Discussion and Method Evaluation 223
References 225
12 Capillary Electromigration Techniques for the Analysis of Drugs and Metabolites in Biological Matrices: A Critical Appraisal 229 Cristiane Masetto de Gaitani, Anderson Rodrigo Moraes de Oliveira, and Pierina Sueli Bonato
12.1 Introduction 229
12.2 Strategies to Obtain Reliable Capillary Electromigration Methods for the Bioanalysis of Drugs and Metabolites 230
12.3 Selected Applications of Capillary Electromigration Techniques in Bioanalysis 238
12.4 Concluding Remarks 243
References 243
13 Capillary Electrophoresis and Multicolor Fluorescent DNA Analysis in an Optofluidic Chip 247 Chaitanya Dongre, Hugo J.W.M. Hoekstra, and Markus Pollnau
13.1 Introduction 247
13.2 Optofluidic Integration in an Electrophoretic Microchip 248
13.3 Fluorescence Monitoring of On-Chip DNA Separation 249
14 Capillary Electrophoresis of Intact Unfractionated Heparin and Related Impurities 267 Robert Weinberger
14.1 Introduction 267
14.2 Capillary Electrophoresis and Heparin 269
14.3 Method Development in Capillary Electrophoresis 269
14.4 Common Impurities Found in Heparin 272
14.5 The United States Pharmacoepia and CE of Heparin 273
14.6 Interlaboratory Collaborative Study 274
14.7 Conclusions 275
References 275
15 Microchip Capillary Electrophoresis for In Situ Planetary Exploration 277 Peter A. Willis and Amanda M. Stockton
15.1 Introduction 277
15.2 Instrument Design 279
15.3 Instrumentation External to the Microdevice 280
15.4 Microdevice Basics 282
15.5 Microdevices and their Applications 285
15.6 Conclusions 289
Acknowledgments 290
References 290
16 Rapid Analysis of Charge Heterogeneity of Monoclonal Antibodies by Capillary Zone Electrophoresis and Imaged Capillary Isoelectric Focusing 293 Yan He, Jim Mo, Xiaoping He, and Margaret Ruesch
16.1 Introduction 293
16.2 Capillary Zone Electrophoresis 295
16.3 Imaged Capillary Isoelectric Focusing 299
16.4 Summary 306
References 307
17 Application of Capillary Electrophoresis for High-Throughput Screening of Drug Metabolism 309 Roman 9Rem?´nek, Jochen Pauwels, Xu Wang, Jos Hoogmartens, Zden9ek Glatz, and Ann Van Schepdael
17.1 Introduction 309
17.2 Sample Deproteinization 310
17.3 On-line Preconcentration 311
17.4 Method Development 312
17.5 Method Validation 314
17.6 Method Applications 315
17.7 Conclusions 316
Acknowledgments 317
References 317
18 Electrokinetic Transport of Microparticles in the Microfluidic Enclosure Domain 319 Qian Liang, Chun Yang, and Jianmin Miao
18.1 Introduction 319
18.2 Numerical Model 320
18.3 Numerical Simulation 322
18.4 Results and Discussion 322
18.5 Model Application 325
18.6 Conclusions 326
References 326
19 Integration of Nanomaterials in Capillary and Microchip Electrophoresis as a Flexible Tool 327 Germa´n A. Messina, Roberto A. Olsina, and Patricia W. Stege
19.1 Introduction 327
19.2 Nanomaterials in Analytical Chemistry 332
19.3 Nanoparticles in Capillary Electrophoresis 333
19.4 Conclusions 352
References 353
20 Microchip Capillary Electrophoresis to Study the Binding of Ligands to Teicoplanin Derivatized on Magnetic Beads 359 Toni Ann Riveros, Roger Lo, Xiaojun Liu, Marisol Salgado, Hector Carmona, and Frank A. Gomez
20.1 Introduction 359
20.2 Experimental Section 359
20.3 Results and Discussion 361
20.4 Conclusions 364
Acknowledgments 365
References 365
21 Glycomic Profiling Through Capillary Electrophoresis and Microchip Capillary Electrophoresis 367 Yehia Mechref
21.1 Introduction 367
21.2 General Considerations of Capillary Electrophoresis and Microchip Capillary Electrophoresis of Glycans 369
21.3 Microchip Capillary Electrophoresis 377
21.4 Conclusions 380
References 381
INDEX 385
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