Capillary Electrophoresis and Microchip Capillary Electrophoresis

Name: Capillary Electrophoresis and Microchip Capillary Electrophoresis | Principles, Applications, and Limitations
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Principles, Applications, and Limitations

Carlos D. García Karin Y. Chumbimuni-Torres Emanuel Carrilho(Autor*in)

Wiley (Verlag)

Erschienen am 1. Februar 2013

416 Seiten

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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 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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Capillary Electrophoresis and Microchip Capillary Electrophoresis: Principles, Applications, and Limitations
Contents
Preface
Acknowledgments
Contributors
1 Critical Evaluation of the Use of Surfactants in Capillary Electrophoresis
1.1 Introduction
1.2 Surfactants for Wall Coatings
1.2.1 Controlling the Electroosmotic Flow
1.2.2 Preventing Adsorption to the Capillary
1.3 Surfactants as Buffer Additives
1.3.1 Micellar Electrokinetic Chromatography
1.3.2 Microemulsion Electrokinetic Chromatography
1.3.3 Nonaqueous Capillary Electrophoresis with Added Surfactants
1.4 Surfactants for Analyte Preconcentration
1.4.1 Sweeping
1.4.2 Transient Trapping
1.4.3 Analyte Focusing by Micelle Collapse
1.4.4 Micelle to Solvent Stacking
1.4.5 Combinations of Preconcentration Methods
1.4.6 Cloud Point Extraction
1.5 Surfactants and Detection in CE
1.5.1 Mass Spectrometry
1.5.2 Electrochemical Detection
1.6 Conclusions
References
2 Sample Stacking: A Versatile Approach for Analyte Enrichment in CE and Microchip-CE
2.1 Introduction
2.2 Isotachophoresis
2.3 Chromatography-Based Sample Stacking
2.4 Methods Based on Electrophoretic Mobility and Velocity Manipulation (Electrophoretic Methods)
2.4.1 Field-Enhanced Sample Stacking (FESS)
2.4.2 Field-Enhanced Sample Injection (FESI)
2.4.3 Large-Volume Sample Stacking (LVSS)
2.4.4 Dynamic pH Junction
2.5 Sample Stacking in Pseudo-Stationary Phases
2.5.1 Field-Enhanced Sample Stacking
2.5.2 Hydrodynamic Injection Techniques
2.5.2.1 Normal Stacking Mode (NSM)
2.5.2.2 Reverse Electrode Polarity Stacking Mode (REPSM)
2.5.2.3 Stacking with Reverse Migrating Micelles (SRMM)
2.5.2.4 Stacking Using Reverse Migrating Micelles and a Water Plug (SRW)
2.5.2.5 High-Conductivity Sample Stacking (HCSS)
2.5.3 Electrokinetic Injection Techniques
2.5.3.1 Field-Enhanced Sample Injection (FESI-MEKC)
2.5.3.2 Field-Enhanced Sample Injection with Reverse Migrating Micelles (FESI-RMM)
2.5.4 Sweeping
2.5.5 Combined Techniques
2.5.5.1 Dynamic pH Junction: Sweeping
2.5.5.2 Selective Exhaustive Injection (SEI)
2.5.6 New Techniques
2.6 Stacking Techniques in Microchips
2.7 Concluding Remarks
References
3 Sampling and Quantitative Analysis in Capillary Electrophoresis
3.1 Introduction
3.2 Injection Techniques in CE
3.2.1 Hydrodynamic Sample Injection
3.2.1.1 Principle
3.2.1.2 Advantages and Performance
3.2.1.3 Disadvantages
3.2.2 Electrokinetic Sample Injection
3.2.2.1 Principle
3.2.2.2 Advantages and Performance
3.2.2.3 Disadvantages
3.2.3 Bias-Free Electrokinetic Injection
3.2.4 Extraneous Sample Introduction Accompanying Injections in CE
3.2.5 Sample Stacking
3.2.5.1 Principle
3.2.5.2 Advantages and Performance
3.2.5.3 Disadvantages
3.2.6 Alternative Batch Sample Injection Techniques
3.2.6.1 Rotary-Type Injectors for CE
3.2.6.2 Hydrodynamic Sample Splitting as Injection Method for CE
3.2.6.3 Electrokinetic Sample Splitting as Injection Method for CE
3.2.6.4 Dual-Opposite End Injection in CE
3.3 Micromachined/Microchip Injection Devices
3.3.1 Droplet Sampler Based on Digital Microfluidics
3.3.2 Wire Loop Injection
3.4 Automated Flow Sample Injection and Hyphenated Systems
3.4.1 Introduction
3.4.2 Advantages and Performance
3.4.3 Disadvantages
3.5 Computerized Sampling and Data Analysis
3.6 Sampling in Portable CE Instrumentation
3.7 Quantitative Analysis in CE
3.7.1 Introduction
3.7.2 Quantitative Analysis with HD Injection
3.7.3 Quantitative Analysis with EK Injection
3.7.4 Validation of the Developed CE Methods
3.7.5 Computer Data Treatment in Quantitative Analysis
3.8 Conclusions
References
4 Practical Considerations for the Design and Implementation of High-Voltage Power Supplies for Capillary and Microchip Capillary Electrophoresis
4.1 Introduction
4.1.1 High-Voltage Fundamentals
4.1.2 Electroosmotic Flow Control
4.1.3 Technical Aspects
4.1.4 Construction of Bipolar HVPS from Unipolar HVPS
4.1.5 Safety Considerations
4.1.6 HVPS Commercially Available
4.1.7 Practical Considerations
4.1.8 Alternative Sources of HV
4.1.9 HVPS Controllers for MCE
4.2 High-Voltage Measurement
4.3 Concluding Remarks
References
5 Artificial Neural Networks in Capillary Electrophoresis
5.1 Introduction
5.2 Optimization in CE: From Single Variable Approach Toward Artificial Neural Networks
5.2.1 Limitations of "Traditional" Single Variable Approach
5.2.2 Multivariate Approach with Experimental Design and Response Surface Modeling
5.2.2.1 Experimental Design
5.2.2.2 Response Surface Modeling
5.3 Artificial Neural Networks in Electromigration Methods
5.3.1 Introduction-Basic Principles of ANN
5.3.2 Optimization Using a Combination of ED and ANN
5.3.2.1 Testing of ED-ANN Algorithm
5.3.2.2 Practical Applications of ED-ANN
5.3.3 Quantitative CE Analysis and Determination from Overlapped Peaks
5.3.3.1 Evaluation of Calibration Plots in CE Using ANN to Increase Precision of Analysis
5.3.3.2 ANN in Quantitative CE Analysis from Overlapped Peaks
5.3.4 ANN in CEC and MEKC
5.3.5 ANN for Peptides Modeling
5.3.6 Classification and Fingerprinting
5.3.7 Other Applications
5.4 Conclusions
Acknowledgments
References
6 Improving the Separation in Microchip Electrophoresis by Surface Modification
6.1 Introduction
6.2 Strategies for Improving Separation
6.2.1 Selection of an Adequate Technique: ME
6.2.2 Microchannel Design
6.2.3 Selection of an Appropriate ME Material
6.2.4 Optimization of the Working Conditions
6.2.5 Surface Modification
6.2.5.1 Surface Micro- and Nanostructuring
6.2.5.2 Employment of Energy Sources
6.2.5.3 Chemical Surface Modification
6.3 Chemical Modifiers
6.3.1 Surfactants
6.3.2 Ionic Liquids
6.3.3 Nanoparticles
6.3.4 Polymers
6.4 Conclusions
Acknowledgments
References
7 Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor: Dissociation Kinetic Analysis Method for "Complexes" Using Capillary Electrophoretic Separation Process
7.1 Introduction
7.2 Basic Concept of CER
7.3 Dissociation Kinetic Analysis of Metal Complexes Using a CER
7.3.1 Determination of the Rate Constants of Dissociation of 1:2 Complexes of Al3+ and Ga3+ with an Azo Dye Ligand 2,2'-Dihydroxyazobenzene-5,5'-Disulfonate in a CER
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 (µCER)
7.5 Expanding the Scope of the CER to the Measurement of Slow Dissociation Kinetics with a Half-Life of Hours
7.5.1 Principle of LS-CER
7.5.2 Application of LS-CER to the Ti(IV)-Catechin Complex
7.5.3 Application of LS-CER to the Ti(IV)-Tiron Complex
7.6 Expanding the Scope of CER to Measurement of the Dissociation Kinetics of Biomolecular Complexes
7.6.1 Dissociation Kinetic Analysis of [SSB-ssDNA] Using CER
7.7 Conclusions
References
8 Capacitively Coupled Contactless Conductivity Detection (C4D) Applied to Capillary Electrophoresis (CE) and Microchip Electrophoresis (MCE)
8.1 Introduction
8.2 Theory of C4D
8.2.1 Basic Principles of C4D
8.2.2 Simulation
8.2.3 Basic Equation for Sensitivity
8.2.4 Equivalent Circuit of a CE-C4D System
8.2.5 Practical Guidelines
8.3 C4D Applied to Capillary Electrophoresis
8.3.1 Instrumental Aspects in CE
8.3.2 Coupling C4D with UV-Vis Photometric Detectors in CE
8.3.3 Fundamental Studies in Capillary Electrophoresis Using C4D
8.3.4 Fundamental Studies on C4D
8.3.5 Applications
8.4 C4D Applied to Microchip Capillary Electrophoresis
8.4.1 Geometry of the Detection Electrodes
8.4.1.1 Embedded Electrodes
8.4.1.2 Attached Electrodes
8.4.1.3 External Electrodes
8.4.2 Applications
8.4.2.1 Bioanalytical Applications
8.4.2.2 On-Chip Enzymatic Reactions
8.4.2.3 Food Analysis
8.4.2.4 Explosives and Chemical Warfare Agents
8.4.2.5 Other Applications
8.5 Concluding Remarks
Acknowledgments
References
9 Capillary Electrophoresis with Electrochemical Detection
9.1 Principles of Electrochemical Detection
9.1.1 Amperometric Detection
9.1.2 Potentiometric Detection
9.1.3 Conductivity Detection
9.2 Interfacing Amperometric Detection to Capillary Electrophoresis
9.2.1 Off-Column Detection
9.2.2 End-Column Detection
9.2.3 Use of Multiple Detection Electrodes
9.2.4 Pulsed Amperometric Detection
9.2.5 Nonaqueous EC Detection
9.2.6 Electrode Material
9.2.7 Dual Conductivity and Amperometric Detection
9.3 Interfacing Electrochemical Detection to Microfluidic Capillary Electrophoresis
9.3.1 End-Column Detection
9.3.2 Pulsed Amperometric Detection
9.3.3 Off-Channel Detection
9.3.4 Electrode Material
9.3.5 Portable CE and MCE Systems
9.3.6 Applications of CE-MCE with AD
9.3.7 Future Directions for CE-MCE with EC Detection
References
10 Overcoming Challenges in Using Microchip Electrophoresis for Extended Monitoring Applications
10.1 Introduction
10.2 Background Electrolyte (BGE) Longevity
10.3 Achieving Rapid Sequential Injections
10.4 Robust Quantitation
10.5 Conclusions
References
11 Distinction of Coexisting Protein Conformations by Capillary Electrophoresis
11.1 Introduction
11.1.1 Theoretical Aspects of in vivo Protein Folding
11.2 Protein Misfolding and Induction of Unfolding
11.3 Conformational Pathologies
11.4 Distinction Between Conformations
11.5 Relevance of Conformations for Biotechnological Products
11.6 Conformational Elucidation-An Overview of Alternative Methods to CE
11.7 HPLC in Conformational Distinction
11.7.1 Intact Proteins
11.7.1.1 Reversed-Phase (RP)-HPLC
11.7.1.2 Size Exclusion (SEC)-HPLC
11.7.1.3 Ion-Exchange-HPLC
11.7.2 HPLC with Detectors Sensitive for Conformations and Aggregates
11.7.3 Peptides as Model Compounds for Hydrophobic Stationary Phases in HPLC
11.8 Capillary Electrophoresis (CE) in Conformational Separations
11.8.1 Fundamental Aspects and Survey of Pitfalls
11.8.2 Electrophoretic Mobility of Proteins
11.8.3 Peak Profiles and Derivable Thermodynamic Aspects of Protein Re-/Unfolding
11.8.4 Dipeptides as a Case Study for Isomerization
11.8.5 Denaturation Factors and Strategies Applied in CE
11.8.5.1 Separation Electrolyte, Injection Solution, and Sample Storage
11.8.5.2 Denaturation by Urea, Dithiothreitol, and GdmCl
11.8.5.3 Effects of pH and Organic Solvents
11.8.5.4 Temperature
11.8.5.5 Electrical Field
11.8.5.6 Detergents
11.8.5.7 Ligands and Ions-Case Studies on Potential Amyloidogenic ß2m
11.8.6 ß-Amyloid Peptides
11.8.6.1 Prions
11.9 Comparison Between CE and HPLC
11.10 Conclusive Discussion and Method Evaluation
11.10.1 General Aspects
11.10.2 HPLC
11.10.3 CE
References
12 Capillary Electromigration Techniques for the Analysis of Drugs and Metabolites in Biological Matrices: A Critical Appraisal
12.1 Introduction
12.2 Strategies to Obtain Reliable Capillary Electromigration Methods for the Bioanalysis of Drugs and Metabolites
12.2.1 Selectivity and Detectability
12.2.1.1 Efficiency
12.2.1.2 Sample Preparation
12.2.1.3 Detectors
12.2.2 Repeatability
12.3 Selected Applications of Capillary Electromigration Techniques in Bioanalysis
12.3.1 Pharmacokinetics and Metabolism Studies
12.3.2 Enantioselective Analysis of Drugs and Metabolites
12.3.3 Biopharmaceuticals or Biotechnology-Derived Pharmaceuticals
12.3.4 Therapeutic Drug Monitoring
12.3.5 Clinical and Forensic Toxicology
12.4 Concluding Remarks
References
13 Capillary Electrophoresis and Multicolor Fluorescent DNA Analysis in an Optofluidic Chip
13.1 Introduction
13.2 Optofluidic Integration in an Electrophoretic Microchip
13.2.1 Sample Fabrication
13.2.2 Optofluidic Characterization
13.3 Fluorescence Monitoring of On-Chip DNA Separation
13.3.1 Experimental Materials and Methods
13.3.2 Experimental Results and Analysis
13.4 Toward Ultrasensitive Fluorescence Detection
13.4.1 Optimization of the Experimental Setup
13.4.2 All-Numerical Postprocessed Noise Filtering
13.5 Multicolor Fluorescent DNA Analysis
13.5.1 Dual-Point, Dual-Wavelength Fluorescence Monitoring
13.5.2 Modulation-Frequency Encoded Multiwavelength Fluorescence Sensing
13.5.3 Application to Multiplex Ligation-Dependent Probe Amplification
13.6 Conclusions and Outlook
Acknowledgments
References
14 Capillary Electrophoresis of Intact Unfractionated Heparin and Related Impurities
14.1 Introduction
14.2 Capillary Electrophoresis and Heparin
14.3 Method Development in Capillary Electrophoresis
14.4 Common Impurities Found in Heparin
14.5 The United States Pharmacoepia and CE of Heparin
14.6 Interlaboratory Collaborative Study
14.7 Conclusions
References
15 Microchip Capillary Electrophoresis for In Situ Planetary Exploration
15.1 Introduction
15.2 Instrument Design
15.3 Instrumentation External to the Microdevice
15.4 Microdevice Basics
15.4.1 All-Glass Devices for Microchip Capillary Electrophoresis
15.4.2 Three-Layer Hybrid Substrate Glass-PDMS Devices for Fluidic Manipulation
15.4.3 Integrating Fluidic Manipulation with Electrophoresis
15.5 Microdevices and their Applications
15.5.1 Microdevices with Bus-Valve Control of Microfluidic Manipulation
15.5.2 Automaton Devices for Programmable Microfluidic Manipulation
15.6 Conclusions
Acknowledgments
References
16 Rapid Analysis of Charge Heterogeneity of Monoclonal Antibodies by Capillary Zone Electrophoresis and Imaged Capillary Isoelectric Focusing
16.1 Introduction
16.2 Capillary Zone Electrophoresis
16.2.1 Separation and Detection Strategy
16.2.1.1 Capillary Construction
16.2.1.2 Buffer Composition
16.2.1.3 Separation Voltage and Field Strength
16.2.1.4 Detection
16.2.2 Applications
16.3 Imaged Capillary Isoelectric Focusing
16.3.1 Method Development and Optimization
16.3.1.1 Carrier Ampholyte
16.3.1.2 Additives
16.3.1.3 Focusing Time and Voltage
16.3.1.4 Salt Concentration
16.3.1.5 Protein Concentration
16.3.2 iCE Method Validation
16.3.3 Applications
16.3.3.1 Cell Line Development Support
16.3.3.2 Formulation Screening
16.3.3.3 Characterization of Acidic Species
16.4 Summary
References
17 Application of Capillary Electrophoresis for High-Throughput Screening of Drug Metabolism
17.1 Introduction
17.2 Sample Deproteinization
17.3 On-line Preconcentration
17.4 Method Development
17.4.1 Dynamic Coating of Inner Capillary Wall
17.4.2 Short-End Injection
17.4.3 Strong Rinsing Procedure
17.4.4 Optimized Method
17.5 Method Validation
17.6 Method Applications
17.6.1 Drug Stability Screening
17.6.2 Kinetic Study
17.7 Conclusions
Acknowledgments
References
18 Electrokinetic Transport of Microparticles in the Microfluidic Enclosure Domain
18.1 Introduction
18.2 Numerical Model
18.2.1 Problem Description
18.2.2 Mathematical Model
18.3 Numerical Simulation
18.4 Results and Discussion
18.4.1 Particle Transport in the Bulk Flow
18.4.1.1 The Particle Velocity in the Confined Domain
18.4.1.2 The Trajectory of Particle Transport within the Confined Domain
18.4.1.3 The Effect of Sidewall Zeta Potential on the Particle Motion
18.4.2 Particle Transport Near the Bottom Surface
18.4.2.1 The Effect of the EDLThickness on the Near Wall Motion of the Particle
18.4.2.2 The Effect of Surface Charge on the Near Wall Transport of the Particle
18.5 Model Application
18.6 Conclusions
References
19 Integration of Nanomaterials in Capillary and Microchip Electrophoresis as a Flexible Tool
19.1 Introduction
19.1.1 Historical Overview of Nanotechnology
19.1.2 Nanomaterials
19.1.2.1 Carbon-Based Nanomaterials
19.1.2.2 Metal-Based Nanomaterials
19.1.2.3 Dendrimers
19.1.2.4 Composites
19.2 Nanomaterials in Analytical Chemistry
19.3 Nanoparticles in Capillary Electrophoresis
19.3.1 Nanoparticles in Capillary Electrochromatography
19.3.1.1 Organic Nanoparticles
19.3.1.2 Inorganic Particles
19.3.2 Nanoparticles in Electrokinetic Chromatography
19.3.2.1 Organic Nanoparticles
19.3.2.2 Inorganic Particles
19.3.3 Nanoparticles in Microchip Electrochromatography
19.4 Conclusions
References
20 Microchip Capillary Electrophoresis to Study the Binding of Ligands to Teicoplanin Derivatized on Magnetic Beads
20.1 Introduction
20.2 Experimental Section
20.2.1 Materials and Methods
20.2.1.1 Equipment and Fabrication of the Microchips
20.2.1.2 Surface Coating
20.2.1.3 Teic Immobilization on Magnetic Microbeads
20.2.2 Procedures
20.2.2.1 FAMCE Studies
20.2.2.2 MFAC Studies
20.3 Results and Discussion
20.3.1 FAMCE Studies
20.3.1.1 Nonspecific Adsorption Resistance
20.3.1.2 The Binding of DA3 to Teic-Beads
20.3.2 MFAC Studies
20.4 Conclusions
Acknowledgments
References
21 Glycomic Profiling Through Capillary Electrophoresis and Microchip Capillary Electrophoresis
21.1 Introduction
21.1.1 Release of N-Glycans from Glycoproteins
21.1.1.1 Chemical Release
21.1.1.2 Enzymatic Release
21.1.2 Release of O-Glycans from Glycoproteins
21.1.2.1 Chemical Release
21.1.2.2 Enzymatic Release
21.2 General Considerations of Capillary Electrophoresis and Microchip Capillary Electrophoresis of Glycans
21.2.1 Capillary Electrophoresis-Laser-Induced Fluorescence (CE-LIF) Analysis of Glycans
21.2.2 Interfacing Capillary Electrophoresis and Capillary Electrochromatography to Mass Spectrometry
21.2.2.1 ESI Interfaces for Capillary Electrophoresis
21.2.2.2 Sheathless-Flow Interface
21.2.2.3 Sheath-Flow Interface
21.2.2.4 Liquid Junction Interface
21.2.2.5 MALDI Interfaces for Capillary Electrophoresis
21.2.2.6 CE-MS Analysis of Glycans
21.2.2.7 Glycomic Analysis by CEC-MS
21.3 Microchip Capillary Electrophoresis
21.4 Conclusions
References
Index

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