EWINGS

ANALYTICAL
INSTRUMENTATION
HANDBOOK
THIRD EDITION

Copyright 2005 by Marcel Dekker

 EWINGS
ANALYTICAL
INSTRUMENTATION
HANDBOOK
THIRD EDITION

Edited by Jack Cazes

Florida Atlantic University
Boca Raton, Florida, U.S.A.

M ARCEL D EKKER N EW Y ORK

Copyright 2005 by Marcel Dekker

Although great care has been taken to provide accurate and current information, neither the author(s) nor the publisher, nor anyone else associated
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Copyright 2005 by Marcel Dekker

Preface to the Third Edition

The Analytical Instrumentation Handbook serves as a guide for workers in analytical chemistry, who need a starting place
for information about a specific instrumental technique, either as a basic introduction to it or as a means of finding leading
references dealing with theory and methodology for an instrumental technique.
The chapters which appeared in the second edition have been thoroughly expanded and updated, with concepts, appli-
cations, and key references to the recent literature. Only one chapter (Laboratory Balances) has been eliminated; eight new
chapters have been added to the Handbook, dealing with
. Microchip technology
. Biosensor technology
. Validation of chromatographic methods
. Gel permeation and size exclusion chromatography
. Field-flow fractionation
. Countercurrent chromatography
. Hyphenated techniques in chromatography, including LC-MS, LC-NMR, and so on
. Thin-layer chromatography
The chapters have been written from the standpoint of the instrumentation as it is in use today, with an introductory
description of the technique(s), and a theoretical treatment of the science and technology, wherever it is applicable or
where it will facilitate an understanding of the instrumentation. However, the major emphasis is on the instrumentation.
The chapters are not, simply, a catalog of commercially available instruments. Nevertheless, in some cases, commercially
available instruments have been used as examples to illustrate design features discussed in the text.
It is sincerely intended and anticipated that this third revised and expanded edition of the Analytical Instrumentation
Handbook will serve as a ready-reference on the desks of all practitioners of instrumental analytical chemistry.

iii

Copyright 2005 by Marcel Dekker

Contents

Preface to the Third Edition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii

Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxi

1. The Laboratory Use of Computers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

Wes Schafer, Zhihao Lin
I. Introduction . . . . 1
II. Computer Components and Design Considerations . . . . 1
A. Motherboard/Central Processing Unit . . . . 2
B. Memory/Cache . . . . 3
C. Disk Storage . . . . 3
D. Video (Graphics Card and Monitor) . . . . 4
E. Other Peripherals . . . . 5
III. Computer Maintenance . . . . 5
A. Performance Monitoring . . . . 5
B. Virus Protection . . . . 6
C. Backup and Recovery . . . . 6
IV. Data Transfer/Instrument Interfaces . . . . 7
A. Transducers . . . . 7
B. Analog Signal Transmission . . . . 7
C. Analog Signal Filtering . . . . 7
D. Analog-to-Digital Conversion . . . . 8
E. Digital Signal Transmission . . . . 10
F. Digital Data Filtering . . . . 11
V. Data Analysis/Chemometrics . . . . 12
A. Multivariate Calibration . . . . 12
B. Pattern Recognition . . . . 16
VI. Data Organization and Storage . . . . 18
A. Automated Data Storage . . . . 19
B. Laboratory Information Management Systems . . . . 19
Bibliography . . . . 20

2. Flow Injection/Sequential Injection Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Elo Harald Hansen, Jianhua Wang
I. Introduction . . . . 21
II. Flow Injection Analysis . . . . 23

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vi Contents

A. Basic Principles . . . . 23
B. Basic FIA Instrumentation . . . . 23
C. Dispersion in FIA . . . . 24
D. Selected Examples to Illustrate the Unique Features of FIA . . . . 25
E. FIA Gradient TechniquesExploiting the Physical Dispersion Process . . . . 30
F. FIA for Process Analysis/Monitoring . . . . 32
III. Further Developments of FIA . . . . 32
A. Sequential Injection Analysis . . . . 33
B. Lab-on-Valve . . . . 33
IV. Selected Applications of FIA/SIA and their Hyphenations in On-Line Sample Pretreatments . . . . 34
A. Separation and Preconcentration Procedures Based on Column Reactors or KRs . . . . 36
B. FIA/SIA On-Line Solvent Extraction and Back Extraction Preconcentration . . . . 39
C. On-Line Hydride/Vapor Generation Preconcentration Schemes . . . . 44
V. Applications of SI-LOV for On-Line Sample Pretreatments . . . . 45
A. The Third Generation of Flow Injection and Micro Total Analysis Systems . . . . 45
B. Applications of mSI-LOV Systems for On-Line Sample Pretreatments with Optical Monitoring . . . . 46
C. SI-BI-LOV: Solving the Dilemma of On-Line Column Separation/Procencentration Schemes . . . . 47
References . . . . 52

3. Inductively Coupled Plasma Optical Emission Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57

Tiebang Wang
I. Introduction . . . . 57
II. An Overview . . . . 59
A. Principle of Atomic Emission Spectrometry . . . . 59
B. Atomic Spectroscopic Sources . . . . 60
C. Atomic Spectroscopic Techniques and Instruments . . . . 60
III. Inductively Coupled Plasma . . . . 61
A. The Discharge . . . . 61
B. Emission Detection and Processing . . . . 62
C. Analytical Performance and General Characteristics . . . . 62
IV. ICP-OES Instrumentation . . . . 63
A. Nebulizers . . . . 63
B. Spray Chambers . . . . 65
C. Alternative Sample Introduction Systems . . . . 65
D. Optics and the Spectrometer . . . . 66
E. Emission Detectors . . . . 69
F. Radially Viewed ICP . . . . 71
V. Applications of ICP-OES . . . . 71
A. Geoanalysis . . . . 71
B. Metallurgical Samples . . . . 72
C. Agricultural and Food Samples . . . . 72
D. Biological and Clinical Samples . . . . 72
E. Environmental Samples . . . . 73
F. Organics . . . . 73
G. Nuclear Materials . . . . 73
VI. Summary and Future Prognosis . . . . 74
References . . . . 74

4. Atomic Absorption Spectrometry and Related Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

Bernhard Welz, Maria Goreti R. Vale
I. Introduction . . . . 75
II. The History of Atomic Spectroscopy . . . . 76
A. The Early History . . . . 76
B. Sir Alan Walsh and the Rebirth of AAS . . . . 77
C. Boris Lvov and the Graphite Furnace . . . . 77

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Contents vii

III. Atomic Spectra in Absorption, Emission, and Fluorescence . . . . 79

A. Origin and Characteristics of Atomic Spectra . . . . 79
B. Line Width and Line Profile . . . . 80
C. Measuring the Absorptionthe Beer Lambert Law . . . . 81
D. The Zeeman Effect . . . . 82
IV. Spectrometers for AAS . . . . 82
A. Medium-Resolution LSAAS . . . . 83
B. High-Resolution CSAAS . . . . 92
V. The Techniques of Atomic Spectrometry . . . . 95
A. The FAAS Technique . . . . 95
B. The GFAAS Technique . . . . 101
C. Chemical Vapor Generation . . . . 112
D. Spectrometers for AFS . . . . 117
E. Spectrometers for Flame OES . . . . 118
VI. Measurement, Calibration, and Evaluation in AAS, AFS, and OES . . . . 118
A. Samples and Measurement Solutions . . . . 118
B. Calibration . . . . 119
C. Evaluation . . . . 121
References . . . . 123

5. Ultraviolet, Visible, Near-Infrared Spectrophotometers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127

Chris W. Brown
I. Introduction . . . . 127
A. Beers Law . . . . 127
B. Deviations from Beers Law . . . . 128
II. Spectrophotometer Characteristics . . . . 129
A. The Architecture of a Spectrophotometer . . . . 131
III. Present and Future UV VIS NIR Spectrometers . . . . 137
References . . . . 139

6. Molecular Fluorescence and Phosphorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141

Fernando M. Lancas, Emanuel Carrilho
I. Introduction . . . . 141
II. Theory . . . . 141
A. Excited-State Processes . . . . 142
B. Excited-State Lifetimes . . . . 142
C. Quantum Yield . . . . 143
D. Quenching . . . . 143
E. Intensity and Concentration . . . . 143
F. Luminescence Spectra . . . . 144
G. Polarization . . . . 144
III. Instrumentation . . . . 145
A. Instrument Components . . . . 145
B. Instrument Configurations for Conventional Fluorescence Measurements . . . . 147
C. Phosphorimeters . . . . 148
D. Instruments with Special Capabilities . . . . 150
IV. Practical Considerations and Applications . . . . 153
A. Environmental Effects . . . . 155
B. Wavelength Calibration and Spectral Correction . . . . 156
C. Background and Interfering Signals . . . . 156
D. Direct Quantitative and Qualitative Analyses . . . . 156
E. Low Temperature Techniques . . . . 156
F. Room-Temperature Phosphorescence . . . . 157
G. Techniques and Applications . . . . 157
H. Future Outlook . . . . 158
References . . . . 159

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viii Contents

7. Vibrational Spectroscopy: Instrumentation for Infrared and Raman Spectroscopy . . . . . . . . . . . . . . . . . 163

Peter Fredericks, Llewellyn Rintoul, John Coates
I. Introduction . . . . 163
A. Background for Vibrational Spectroscopic Measurements . . . . 164
B. The Basic Principles of Vibrational Spectroscopy . . . . 166
C. Comparison of Techniques and Relative Roles . . . . 168
II. IR Instrumentation . . . . 170
A. Types of Instruments and Recent Trends . . . . 170
B. Instrumentation: Design and Performance Criteria . . . . 172
C. NIR Instrumentation . . . . 206
D. FIR (Terahertz) Instrumentation . . . . 209
III. Raman Instrumentation . . . . 211
A. Types of Instrumentation and Recent Trends . . . . 211
B. Instrumentation: Design and Performance Criteria . . . . 213
IV. Practical Issues of Implementation of IR and Raman Instruments . . . . 230
A. Instrument Architecture and Packaging . . . . 230
B. Data Handling and Computer Applications . . . . 232
V. Instrumentation Standards . . . . 233
VI. Recommended Reference Sources . . . . 234
Suggested Reference Texts . . . . 234
General Texts and Reviews . . . . 235
Historical Reviews . . . . 235
References . . . . 235

8. X-Ray Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239

Narayan Variankaval
I. Introduction . . . . 239
A. Continuous and Characteristic Spectra . . . . 239
B. Diffraction from Crystals . . . . 240
C. Auger Effect . . . . 241
D. Wavelength Dispersive X-Ray Spectrometry . . . . 241
E. Energy Dispersive X-Ray Spectrometry . . . . 241
F. X-Ray Reflectivity . . . . 241
G. X-Ray Absorption . . . . 241
II. Instrumentation and Methods . . . . 242
A. Overview of Optical Components of an X-Ray Diffractometer . . . . 242
B. X-Ray Methods . . . . 247
III. Concluding Remarks . . . . 255
Additional Reading . . . . 255
General . . . . 255
X-Ray Properties of Elements . . . . 255
X-Ray Diffraction . . . . 255
X-Ray Detectors . . . . 255
Particle Induced X-Ray Emission . . . . 255
X-Ray Fluorescence . . . . 255
X-Ray Photoelectron Spectroscopy . . . . 255
X-Ray Reflection . . . . 255
X-Ray Microanalysis . . . . 255
References . . . . 255

9. Photoacoustic Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 257

A.K. Rai, S.N. Thakur, J.P. Singh
I. Introduction . . . . 257
II. Photoacoustics and PA Spectroscopy . . . . 258
A. History . . . . 258
B. Principle . . . . 259

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Contents ix

III. Relevance of PA to Plant Science . . . . 261

IV. Review of PA Applications in Plant Sciences . . . . 262
V. PA in Detection of Plant Disease . . . . 263
A. Experimental Setup . . . . 263
B. Diseases in Wheat Plants . . . . 263
C. Wheat Genotype EKBSN-1 AND FBPFM-2 . . . . 267
D. Virus Disease in Mung Bean Plant . . . . 267
E. Fungal Disease in Sugarcane . . . . 268
F. Virus Disease in Okra Plants . . . . 268
G. Seed-Borne Diseases in Wheat and Rice . . . . 268
VI. Conclusion . . . . 268
VII. Future Developments . . . . 268
References . . . . 269

10. Techniques of Chiroptical Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271

Harry G. Brittain, Nelu Grinberg
I. Introduction to Chiroptical Phenomena . . . . 271
II. Polarization Properties of Light . . . . 272
III. Optical Rotation and ORD . . . . 272
A. Applications of Optical Rotation and ORD . . . . 275
IV. Circular Dichroism . . . . 276
A. Applications of CD . . . . 277
V. CPL Spectroscopy . . . . 278
A. Applications of CPL Spectroscopy . . . . 280
VI. Vibrational Optical Activity . . . . 283
A. Vibrational Circular Dichroism . . . . 283
B. Raman Optical Activity . . . . 285
VII. Fluorescence Detected Circular Dichroism . . . . 288
A. Applications of FDCD . . . . 289
VIII. Concluding Remarks . . . . 290
References . . . . 290

11. Nuclear Magnetic Resonance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295

Frederick G. Vogt
I. Introduction . . . . 295
II. Instrument Design . . . . 296
A. Magnet Systems . . . . 296
B. NMR Probes . . . . 300
C. RF Generation and Signal Detection . . . . 303
D. Magnetic Field Gradients . . . . 306
E. Computer Systems . . . . 306
F. Accessories . . . . 307
III. Theoretical Background . . . . 307
A. Nuclear Spin Dynamics . . . . 307
B. External Manipulations of Spin Coherence . . . . 310
C. Internal Spin Interactions . . . . 311
D. Relaxation Phenomena and Chemical Dynamics . . . . 314
IV. Experimental Methods . . . . 317
A. Basic Pulse Methods . . . . 317
B. Multidimensional NMR Spectroscopy . . . . 322
C. Multiple Resonance and Heteronuclear Techniques . . . . 325
D. Diffusion, Dynamics, and Relaxation Measurements . . . . 332
V. Data Analysis and Interpretation . . . . 334
A. Spectral Processing . . . . 335
B. Manual Data Interpretation . . . . 337

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x Contents

C. Predictive Methods . . . . 342

D. Computer-Assisted Structure Elucidation . . . . 343
VI. Conclusion . . . . 343
References . . . . 344

12. Electron Paramagnetic Resonance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349

Sandra S. Eaton, Gareth R. Eaton
I. Introduction . . . . 349
A. EPR Experiment . . . . 350
B. CW Spectroscopy . . . . 352
C. Pulse Spectroscopy . . . . 355
D. Multiple Resonance Methods . . . . 355
II. What One Can Learn From an EPR Measurement . . . . 357
A. Is There an EPR Signal? Under What Conditions? . . . . 357
B. Lineshape . . . . 358
C. g-Value . . . . 358
D. Spin Spin Coupling . . . . 360
E. Relaxation Times . . . . 360
F. Saturation Behavior . . . . 361
G. Signal Intensity . . . . 362
III. EPR Spectrometer . . . . 363
A. Microwave System . . . . 363
B. EPR Resonators . . . . 366
C. Magnet System . . . . 370
D. Signal Detection and Amplification . . . . 372
E. Data System . . . . 373
F. Commercial EPR Spectrometers . . . . 373
IV. The Sample . . . . 374
A. Spin System . . . . 374
B. Temperature . . . . 374
C. Amount of Sample . . . . 375
D. Phase of Sample . . . . 376
E. Impurities, Overlapping Spectra . . . . 377
F. Intermolecular vs. Intramolecular Interactions . . . . 377
G. Other Environmental Effects . . . . 377
V. Quantitative Measurements of Spin Density . . . . 378
A. Spectrometer Calibration . . . . 378
B. Calibration of Sample Tubes . . . . 379
C. Consideration of Cavity Q . . . . 379
D. Reference Samples for Quantitative EPR . . . . 380
E. Scaling Results for Quantitative Comparisons . . . . 380
VI. Guidance on Experimental Technique . . . . 381
A. Words of Caution . . . . 381
B. Selection of Operating Conditions . . . . 382
C. Second Derivative Operation . . . . 384
D. CW Saturation . . . . 384
E. Methods of Measuring Relaxation Times . . . . 384
F. Measurement of B1 . . . . 385
G. Line vs. Point Samples . . . . 386
H. Overcoupled Resonators . . . . 386
I. The ESE Flip Angle for High-Spin Systems . . . . 387
VII. Less Common Measurements with EPR Spectrometers . . . . 387
A. Saturation-Transfer Spectroscopy . . . . 387
B. Electrical Conductivity . . . . 388
C. Static Magnetization . . . . 388
D. EPR Imaging . . . . 388
E. Pulsed Magnetic Field Gradients . . . . 388

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Contents xi

VIII. Reporting Results . . . . 388

A. Reporting Experimental Spectra . . . . 389
B. Reporting Derived Values . . . . 389
References . . . . 390

13. X-Ray Photoelectron and Auger Electron Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399

C. R. Brundle, J. F. Watts, J. Wolstenholme
I. Introduction . . . . 399
A. X-Ray Photoelectron Spectroscopy . . . . 400
B. Auger Electron Spectroscopy . . . . 401
C. Depth Analyzed for Solids by Electron Spectroscopy . . . . 402
D. Comparison of XPS and AES . . . . 403
II. Electron Spectrometer Design . . . . 404
A. The Vacuum System . . . . 404
B. The Specimen and Its Manipulation . . . . 404
C. X-Ray Sources for XPS . . . . 405
D. Charge Compensation in XPS . . . . 406
E. The Electron Gun for AES . . . . 407
F. Electron Energy Analyzers for Electron Spectroscopy . . . . 408
G. Detectors . . . . 410
H. Spectrometer Operation for Small Area XPS . . . . 411
I. XPS Imaging and Mapping . . . . 411
J. Angle Resolved XPS . . . . 412
III. Interpretation of Photoelectron and Auger Spectra . . . . 413
A. Qualitative Analysis . . . . 413
B. Chemical State Information: The Chemical Shift . . . . 414
C. Chemical State Information: Fine Structure Associated with Core Level Peaks . . . . 417
D. Quantitative Analysis . . . . 418
IV. Compositional Depth Profiling . . . . 420
A. Angle Resolved X-Ray Photoelectron Spectroscopy . . . . 420
B. Variation of Electron Kinetic Energy . . . . 423
C. Ion Sputter Depth Profiling . . . . 424
D. Summary of Depth Profiling . . . . 425
V. Areas of Application and Other Methods . . . . 426
References . . . . 426

14. Mass Spectrometry Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 429

Li-Rong Yu, Thomas P. Conrads, Timothy D. Veenstra
I. Introduction . . . . 429
II. Ionization Methods . . . . 430
A. Matrix-Assisted Laser Desorption/Ionization . . . . 430
B. Electrospray Ionization . . . . 431
III. Mass Analyzers . . . . 434
A. Triple Quadrupole Mass Spectrometer . . . . 434
B. Time-of-Flight Mass Spectrometer . . . . 435
C. Quadrupole Time-of-Flight Mass Spectrometer . . . . 435
D. Ion-Trap Mass Spectrometer . . . . 436
E. Fourier Transform Ion Cyclotron ResonanceMass Spectrometry . . . . 437
F. Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry . . . . 439
G. Inductively Coupled Plasma Mass Spectrometry . . . . 440
IV. Conclusions . . . . 441
References . . . . 441

15. Thermoanalytical Instrumentation and Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445

Kenneth S. Alexander, Alan T. Riga, Peter J. Haines
I. Introduction . . . . 445
A. Scope of Thermal Analysis . . . . 445

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xii Contents

B. Nomenclature and Definitions . . . . 446

C. Symbols . . . . 448
II. Thermogravimetry . . . . 449
A. Development . . . . 449
B. Design Factors . . . . 449
C. The Balance . . . . 450
D. Furnaces and Temperature . . . . 451
E. Computer Workstations to Record, Process, and Control the Acquisition of Data . . . . 452
F. Reporting Results . . . . 453
G. Standards for TG . . . . 454
III. TGA and DTG . . . . 456
IV. DTA and DSC . . . . 457
A. Basic Considerations . . . . 457
B. Design Factors and Theory . . . . 458
C. Experimental Factors . . . . 463
D. Reporting Results . . . . 464
E. Calibration . . . . 465
V. Evolved-Gas Analysis . . . . 467
A. Introduction . . . . 467
B. Instrumentation for EGA . . . . 468
C. The Basic Unit . . . . 469
D. Detector Devices . . . . 469
E. Precursor Considerations . . . . 470
F. Separation Procedures . . . . 470
G. Reporting Data . . . . 471
VI. Thermomechanical Methods . . . . 471
A. Introduction . . . . 471
B. TDA and TMA . . . . 473
C. Dynamic Mechanical Analysis . . . . 473
D. Reporting Data . . . . 474
E. Instrumentation . . . . 474
VII. Dielectric Thermal Analysis . . . . 476
VIII. Calorimetry and Microcalorimetric Measurements . . . . 478
A. Introduction . . . . 478
B. Calorimeters . . . . 482
C. Instrumentation . . . . 482
D. Applications of Isothermal Calorimetry . . . . 486
E. Summary . . . . 490
IX. Other Techniques . . . . 490
X. Simultaneous Thermal Analysis Techniques . . . . 491
A. The Rationale Behind the Simultaneous Approach . . . . 491
B. Simultaneous TG DTA . . . . 491
C. Simultaneous TG DSC . . . . 492
XI. Other Less-Common Techniques . . . . 493
A. Optical Techniques . . . . 493
B. Spectroscopic Techniques . . . . 493
C. X-Ray Techniques . . . . 494
D. DTA Rheometry . . . . 494
E. Microthermal Analysis . . . . 494
F. Applications for TG DTA . . . . 495
G. Applications of TG DSC . . . . 496
XII. Applications . . . . 497
A. Nature of the Solid State . . . . 497
B. Applications Based on Thermodynamic Factors . . . . 498
C. Applications Based on Kinetic Features . . . . 500
D. Applications for EGA . . . . 500
References . . . . 503

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Contents xiii

16. Potentiometry: pH and Ion-Selective Electrodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509

Ronita L. Marple, William R. LaCourse
I. Introduction . . . . 509
A. Electrochemical Cells . . . . 509
B. Activity and Activity Coefficients . . . . 510
C. The Nernst Equation . . . . 512
D. Formal Potentials . . . . 512
E. Liquid Junction Potentials . . . . 513
F. Temperature Coefficients . . . . 513
II. Reference Electrodes . . . . 513
A. The Standard Hydrogen Electrode . . . . 514
B. The Calomel Electrode . . . . 514
C. Silver, Silver Chloride Electrode . . . . 515
D. Double Junction Reference Electrode . . . . 515
E. Reference Electrode Practice . . . . 515
III. Indicator Electrodes . . . . 516
A. Metallic Indicator Electrodes . . . . 516
B. Membrane Indicator Electrodes . . . . 516
IV. General Instrumentation . . . . 523
A. Potentiometers . . . . 523
B. Direct Reading Instruments . . . . 523
C. Commercial Instrumentation and Software . . . . 523
V. Applications of Potentiometry . . . . 524
VI. Current Research Activity . . . . 524
A. Increasing Sensitivity, Lowering Detection Limits . . . . 524
B. New Membrane Materials . . . . 525
C. Biosensors . . . . 525
D. Miniaturization . . . . 525
E. Potentiometric Detectors in Fluidic Streams . . . . 525
VII. Conclusions . . . . 526
References . . . . 526

17. Voltammetry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 529

Mark P. Olson, William R. LaCourse
I. Introduction . . . . 529
II. General Instrumentation . . . . 529
III. Oxidation/Reduction . . . . 530
IV. Polarization and iR Drop . . . . 531
V. The Voltammogram . . . . 532
VI. Mass-Transport . . . . 532
VII. The Diffusion Layer . . . . 533
VIII. Faradaic Current . . . . 534
IX. Non-Faradaic Current . . . . 534
X. Voltage/Time/Current Interdependence . . . . 535
XI. Cyclic Voltammetry . . . . 535
XII. Hydrodynamic Voltammetry . . . . 536
XIII. Polarography . . . . 536
XIV. Waveforms . . . . 538
XV. Innovative Applications of Voltammetry . . . . 539
A. Protein Film Voltammetry . . . . 539
B. Voltammetry on Microfluidic Devices . . . . 540
C. Fast-Scan Cyclic Voltammetry . . . . 540
D. Ultramicroelectrodes . . . . 541
E. Sinusoidal Voltammetry . . . . 541

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xiv Contents

XVI. Suppliers of Analytical Instrumentation and Software . . . . 541

XVII. Voltammetry Simulation Software . . . . 542
References . . . . 542

18. Electrochemical Stripping Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 545

William R. LaCourse
I. Introduction . . . . 545
II. Fundamentals of Stripping Analysis . . . . 546
A. Anodic Stripping Voltammetry . . . . 546
B. Cathodic Stripping Voltammetry . . . . 549
C. Stripping Chronopotentiometry . . . . 549
D. Adsorptive Stripping Voltammetry . . . . 550
E. Stripping Tensammetry . . . . 551
F. Experimental Considerations . . . . 551
III. Instrumentation . . . . 552
A. Cells . . . . 552
B. Electrodes . . . . 552
C. Stripping Analysis in Flowing Streams . . . . 554
D. Stripping Analyzers . . . . 555
E. Modeling Software and Speciation . . . . 557
IV. Applications . . . . 557
V. Conclusions . . . . 558
References . . . . 558

19. Measurement of Electrolytic Conductance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 561

Stacy L. Gelhaus, William R. LaCourse
I. Introduction . . . . 561
A. Principles of Conductivity . . . . 562
B. Strong Electrolytes . . . . 563
C. Weak Electrolytes . . . . 563
D. Ion Mobility and Transport . . . . 564
II. Instrumentation . . . . 566
A. Immersed Electrode Measurements . . . . 566
B. Electrodeless (Noncontacting) Measurements . . . . 574
III. Summary . . . . 577
References . . . . 578

20. Microfluidic Lab-on-a-Chip . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 581

Paul C. H. Li, Xiujun Li
I. Introduction . . . . 581
II. Micromachining Methods . . . . 582
A. Micromachining of Silicon . . . . 582
B. Micromachining of Glass . . . . 583
C. Micromachining of Fused Silica (or Fused Quartz) . . . . 587
D. Micromachining of Polymeric Chips . . . . 588
E. Metal patterning . . . . 594
III. Microfluidic Flow . . . . 594
A. EOF and Hydrodynamic Flow . . . . 594
B. Surface Modifications for Flow Control . . . . 595
C. Fraction Collection . . . . 596
D. Laminar Flow for Liquid Extraction and Microfabrication . . . . 596
E. Concentration Gradient Generation . . . . 597

Copyright 2005 by Marcel Dekker

Contents xv

F. Microvalves . . . . 599
G. Micromixers . . . . 600
H. Alternative Pumping Principles . . . . 600
I. Microfluidic Flow Modeling Study . . . . 601
IV. Sample Introduction . . . . 602
A. Electrokinetic Injection . . . . 602
B. Other Sample Injection Methods . . . . 606
V. Sample Preconcentration . . . . 607
A. Sample Stacking . . . . 607
B. Extraction . . . . 608
C. Sample Volume Reduction . . . . 608
D. Other Preconcentration Methods . . . . 610
VI. Separation . . . . 610
A. Capillary Zone Electrophoresis . . . . 611
B. Capillary Gel Electrophoresis . . . . 612
C. Micellar Electrokinetic Capillary Chromatography . . . . 613
D. Derivatizations for CE for Separations . . . . 614
E. Isotachophoresis . . . . 615
F. Capillary Electrochromatography (CEC) . . . . 616
G. Synchronized Cyclic Capillary Electrophoresis . . . . 617
H. Free-Flow Electrophoresis . . . . 617
VII. Detection Methods . . . . 617
A. Optical Detection Methods . . . . 617
B. Electrochemical Detection . . . . 623
C. Mass Spectrometry . . . . 627
VIII. Applications to Cellular Analysis . . . . 627
A. Slit-Type Filters . . . . 627
B. Weir-Type Filters . . . . 631
C. Cell Adhesion . . . . 632
D. Studies of Cells in a Flow . . . . 633
E. DEP for Cell Retention . . . . 634
IX. Applications to DNA Analysis . . . . 635
A. DNA Amplification . . . . 635
B. DNA Hybridization . . . . 638
C. DNA Sequencing . . . . 642
D. High-Throughput DNA Analysis . . . . 642
E. Other DNA Applications . . . . 643
X. Applications to Protein Analysis . . . . 644
A. Immunoassay . . . . 644
B. Protein Separation . . . . 647
C. Enzymatic Assays . . . . 648
D. MS Analysis for Proteins and Peptides . . . . 650
References . . . . 659

21. Biosensor Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 681

Raluca-Ioana Stefan, Jacobus Fredeiczzk van Staden, Hassan Y. Aboul-Enein
I. Introduction . . . . 681
II. Biological Materials . . . . 681
III. Transducers . . . . 682
A. Electrochemical Transducers . . . . 682
B. Optical Transducers . . . . 682
C. Screen-Printed Electrodes . . . . 683
IV. Immobilization Procedures of the Biological Material . . . . 683
A. Physical Immobilization . . . . 683
B. Chemical Immobilization . . . . 684
V. Design of a Biosensor for an Extended Use and Storage Life . . . . 684
VI. Array-Based Biosensors . . . . 684

Copyright 2005 by Marcel Dekker

xvi Contents

VII. Design of Flow Injection Analysis/Biosensors and Sequential Injection

Analysis/Biosensors Systems . . . . 684
A. Flow Injection/Biosensor(s) System . . . . 684
B. Sequential Injection/Biosensor(s) System . . . . 685
References . . . . 685

22. Instrumentation for High-Performance Liquid Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687

Raymond P. W. Scott
I. Introduction to the Chromatographic Process . . . . 687
A. The Plate Theory . . . . 688
B. Molecular Interactions . . . . 690
C. The Thermodynamic Explanation of Retention . . . . 692
D. Control of Retention by Stationary Phase Availability . . . . 693
II. The Basic Chromatograph . . . . 696
A. Solvent Reservoirs . . . . 696
B. Solvent Programers . . . . 697
C. Nonreturn Valves . . . . 699
D. Chromatography Pumps . . . . 699
E. Column Design and Performance . . . . 703
F. Sample Valves . . . . 705
G. Column Ovens . . . . 707
H. The LC Column . . . . 708
I. Detectors . . . . 713
III. The Modern Comprehensive LC System . . . . 724
References . . . . 725

23. Gas Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 727

Mochammad Yuwono, Gunawan Indrayanto
I. Introduction . . . . 727
II. Basic Instrumentation . . . . 728
III. The Separation System . . . . 728
A. Modes of GC . . . . 728
B. Gas Chromatogram . . . . 728
IV. Gas Supply System . . . . 730
A. Selection of Carrier Gases . . . . 730
B. Gas Sources and Purity . . . . 730
V. Gas Chromatographic Columns . . . . 731
A. Packed Columns . . . . 731
B. Capillary Columns . . . . 732
C. Stationary Phases in GC . . . . 732
D. Optimization of the GC Column Parameters . . . . 733
E. Connection Techniques of Capillary Column . . . . 736
VI. Sample Inlets . . . . 736
A. Syringe Handling Techniques . . . . 736
B. Packed Inlet System . . . . 736
C. Split Injection . . . . 737
D. Splitless Injectors . . . . 737
E. Direct Injection . . . . 737
F. Programed-Temperature Injection . . . . 738
G. Injectors for Gas Samples . . . . 738
H. Headspace Analysis . . . . 738
I. Solid Phase Microextraction Headspace GC . . . . 739
J. Purge and Trap GC Systems . . . . 739
K. Pyrolysis GC . . . . 739
VII. Oven . . . . 740
A. Conventional GC-Oven . . . . 740

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Contents xvii

B. Flash GC-Oven . . . . 740

C. Microwave GC-Oven . . . . 740
D. Infrared Heated GC . . . . 740
VIII. Detector . . . . 741
A. Classification and General Properties . . . . 741
B. Flame Ionization Detector . . . . 742
C. Thermal Conductivity Detector . . . . 742
D. Electron-Capture Detector . . . . 742
E. Nitrogen/Phosphorous Detector . . . . 743
F. Flame Photometric Detector . . . . 744
G. Atomic Emission Detection . . . . 744
H. Gas Chromatography-Mass Spectrometry . . . . 744
IX. Multi Dimensional GC and Comprehensive Two-Dimensional GC . . . . 750
A. Multidimensional GC . . . . 750
B. Comprehensive Two-Dimensional GC . . . . 751
X. Chromatography Data System . . . . 752
XI. Qualitative and Quantitative Analyses . . . . 753
A. Qualitative Analysis . . . . 753
B. Quantitative Analysis . . . . 753
XII. Recent Developments . . . . 754
A. Fast GC . . . . 754
B. Portable GC . . . . 755
C. On-Line LC and GC . . . . 755
References . . . . 755

24. Supercritical Fluid Chromatography Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 759

Thomas L. Chester, J. David Pinkston
I. Introduction . . . . 759
A. Intermolecular Forces in SFC . . . . 760
B. Fluids and Phase Behavior . . . . 760
C. Chromatography with Supercritical Fluid Mobile Phases . . . . 764
D. Reasons for Considering SFC . . . . 768
II. Instrumentation . . . . 769
A. Safety Considerations . . . . 769
B. General SFC Instrumentation Features . . . . 770
C. Packed-Column SFC Instrumentation . . . . 770
D. OT-Column SFC Instrumentation . . . . 776
E. Testing for Proper Operation . . . . 784
F. Flow Behavior with Upstream Pressure Control . . . . 784
III. Method Development . . . . 785
A. Packed-Column SFC Method Development . . . . 785
B. OT-SFC Method Development . . . . 789
IV. Supercritical Fluid Chromatography Mass Spectrometry . . . . 791
A. SFC MS Interfaces . . . . 791
B. Type of Mass Spectrometer . . . . 796
V. Solute Derivatization . . . . 796
VI. Chiral SFC . . . . 797
VII. Conclusion . . . . 798
References . . . . 799

25. Capillary Electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 803

Hassan Y. Aboul-Enein, Imran Ali
I. Introduction . . . . 803
II. Theory of CE . . . . 804
III. Modes of CE . . . . 805

Copyright 2005 by Marcel Dekker

xviii Contents

IV. Instrumentation . . . . 806

A. Sample Injection . . . . 806
B. Separation Capillary . . . . 806
C. High-Voltage Power Supply . . . . 807
D. Background Electrolyte . . . . 807
E. Detection . . . . 807
V. Data Integration . . . . 811
VI. Sample Preparation . . . . 812
VII. Method Development and Optimization . . . . 813
VIII. Methods Validation . . . . 813
IX. Application . . . . 813
X. Conclusions . . . . 822
References . . . . 822

26. Gel Permeation and Size Exclusion Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 827

Gregorio R. Meira, Jorge R. Vega, Mariana M. Yossen
I. General Considerations . . . . 827
II. Basic Concepts . . . . 828
A. The Chromatographic System . . . . 828
B. Simple and Complex Polymers . . . . 829
C. Molar Mass Distribution . . . . 829
D. Entropy-Controlled Partitioning . . . . 832
E. Intrinsic Viscosity and Hydrodynamic Volume . . . . 832
F. SEC Resolution and Column Efficiency . . . . 833
III. Concentration Detectors and Molar Mass Calibration . . . . 834
A. Differential Refractometer . . . . 834
B. UV Sensor . . . . 834
C. Molar Mass Calibrations . . . . 835
D. Chemical Composition . . . . 836
IV. Molar Mass Sensitive Detectors . . . . 836
A. LS Detector . . . . 837
B. Viscosity Detector . . . . 838
C. Osmotic Pressure Detector . . . . 839
V. Experimental Difficulties . . . . 839
A. The High Molecular Weight Saturation Problem . . . . 839
B. Enthalpic Interactions . . . . 840
C. Band Broadening . . . . 840
D. Polyelectrolytes . . . . 840
VI. Instrumentation . . . . 841
A. High-Temperature Equipment . . . . 841
B. Solvent Delivery System . . . . 841
C. Sample Preparation and Injection . . . . 843
D. Fractionation Columns . . . . 844
E. Detectors . . . . 855
VII. Application Examples . . . . 862
A. MMD of a Linear PS . . . . 862
B. Chain Branching in PVAc . . . . 863
C. Characterization of Starch . . . . 864
D. Sample Fractionation by Preparative SEC . . . . 865
References . . . . 866

27. Field-Flow Fractionation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 871

Martin E. Schimpf
I. Introduction . . . . 871
II. Principles and Theory of Retention . . . . 872

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Contents xix

A. The Brownian Mode of Retention . . . . 873

B. The Steric/Hyperlayer Mode . . . . 875
III. Experimental Procedures . . . . 876
A. Sample Injection/Relaxation . . . . 876
B. Selectivity . . . . 877
C. Resolution and Fractionating Power . . . . 877
D. Field Programing . . . . 877
E. Ancillary Equipment . . . . 878
F. Optimization . . . . 879
IV. Applications . . . . 879
A. Polymers . . . . 879
B. Proteins and DNA . . . . 881
C. Colloids (Suspended Particles with d , 1 mm) . . . . 883
D. Starch Granules, Cells, and other Micrometer-Sized Particles (d . 1 mm) . . . . 885
E. Environmental Applications . . . . 886
References . . . . 887

28. Instrumentation for Countercurrent Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 893

Yoichiro Ito
I. Introduction . . . . 893
II. Hydrostatic Equilibrium CCC Systems . . . . 895
A. Development of Hydrostatic CCC Schemes . . . . 895
B. Instrumentation of Hydrostatic CCC Schemes . . . . 895
III. Hydrodynamic Equilibrium Systems . . . . 901
A. Rotary-Seal-Free Centrifuge Systems . . . . 902
B. Analysis of Acceleration on Synchronous Planetary Motions . . . . 902
C. Instrumentation . . . . 908
IV. Special Techniques . . . . 927
A. pH-Zone-Refining CCC . . . . 927
B. Affinity CCC . . . . 931
C. CCC/MS . . . . 931
V. Two-Phase Solvent Systems . . . . 931
A. Retention Volume and Partition Coefficient (K) . . . . 931
B. Rs and Retention of the Stationary Phase in CCC . . . . 932
C. Various Parameters of High-Speed CCC Affecting Stationary Phase Retention . . . . 934
D. Partition Coefficient of Samples . . . . 936
VI. Future Instrumentation . . . . 937
A. Analytical CCC . . . . 937
B. Preparative CCC . . . . 937
References . . . . 938

29. HPLC-Hyphenated Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 945

R. A. Shalliker, M. J. Gray
I. Introduction . . . . 945
II. Liquid ChromatographyMass Spectroscopy . . . . 946
A. Manual Treatment . . . . 947
B. Mechanical or Moving Transport Interfaces . . . . 947
C. Fast Atom Bombardment Interfaces . . . . 949
D. Particle Beam and Monodisperse Aerosol Generation Interfaces . . . . 951
E. MALDI and Laser-Assisted Interfaces . . . . 953
F. Atmospheric Pressure Ionization and Nebulizer Based Interfaces . . . . 955
G. Atmospheric Pressure Chemical Ionization Interface . . . . 955
H. Atmospheric Sampling Glow Discharge Ionization Interface . . . . 957
I. Thermospray Ionization Interfaces . . . . 958
J. Atmospheric Pressure Electrospray/Ionspray Ionization Interfaces . . . . 959
K. Sonic Spray Interface . . . . 963

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xx Contents

L. Atmospheric Pressure Spray Ionization . . . . 964

M. Atmospheric Pressure Photoionization Interface . . . . 964
N. Cold Spray Ionization Interface . . . . 965
O. Interchangeable API Interfaces . . . . 965
P. High Throughput Strategies in HPLC-MS . . . . 965
III. Liquid Chromatography Fourier Transform-Infrared Spectroscopy . . . . 966
A. Flow Cell Methods of Analysis . . . . 967
B. Online Removal of Water Prior to Flow Cell Detection . . . . 968
C. Solvent Elimination Methods . . . . 969
D. Deposition Substrates . . . . 969
E. Solvent Elimination Interfaces . . . . 970
F. Thermospray Nebulizer . . . . 970
G. Concentric Flow Nebulizer . . . . 970
H. Pneumatic Nebulizer . . . . 972
I. Ultrasonic Nebulizer . . . . 972
J. Electrospray Nebulizer . . . . 973
K. Quantitative Considerations . . . . 974
IV. Liquid Chromatography Nuclear Magnetic Resonance Spectroscopy . . . . 974
A. Solvent Suppression . . . . 974
B. Sensitivity . . . . 975
C. Trace Enrichment . . . . 977
V. Liquid Chromatography-Inductively Coupled Plasma Spectroscopy . . . . 978
VI. Multiple Hyphenation (or Hypernation) of LC with Spectroscopy . . . . 980
VII. Hyphenation of Liquid Chromatography with Gas Chromatography . . . . 982
A. Concurrent Solvent Evaporation Techniques . . . . 982
B. Partially Concurrent Solvent Evaporation . . . . 983
C. Fully Concurrent Solvent Evaporation Techniques . . . . 985
D. Programmed Temperature Vaporizer/Vaporizer Interfaces . . . . 986
E. Miscellaneous . . . . 988
References . . . . 988

30. Thin Layer Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 995

Joseph Sherma
I. Introduction . . . . 995
II. Sample Preparation . . . . 996
III. Stationary Phases . . . . 997
IV. Mobile Phases . . . . 997
V. Application of Samples . . . . 998
VI. Chromatogram Development . . . . 999
A. Capillary-Flow TLC . . . . 999
B. Forced Flow Planar Chromatography . . . . 1001
VII. Zone Detection . . . . 1003
VIII. Documentation of Chromatograms . . . . 1004
IX. Zone Identification . . . . 1005
X. Quantitative Analysis . . . . 1006
A. Nondensitometric Methods . . . . 1006
B. Densitometric Evaluation . . . . 1007
XI. TLC Combined with Spectrometric Methods . . . . 1009
A. Mass Spectrometry . . . . 1009
B. Infrared Spectrometry . . . . 1009
C. Raman Spectrometry . . . . 1009
XII. Preparative Layer Chromatography . . . . 1009
XIII. Thin Layer Radiochromatography . . . . 1010
XIV. Applications of TLC . . . . 1011
References . . . . 1012

Copyright 2005 by Marcel Dekker