Frans_M_Everaerts_Isotachophoresis_378342.pdf

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CONTENTS IX 6.4.2. The d.c. method of resistance determination ............................. 6.4.3. The d.c.-a.c. converter ............................................. 6.4.4. The a.c. method of resistance determination ............................. 6.4.5. Conductivity probe with equiplanar-mounted sensing electrodes ............. 6.5. UV absorption meter .................................................... 6.5.1. Introduction ..................................................... 6.5.2. Construction of the UV source ................................... 6.5.3. UV detector in combination with a non-modulated UV source ............... 6.5.4. UV detector in combination with a modulated UV source .................. 6.5.5.UVcell ......................................................... 6.5.6. Experimental .................................................... 6.6. Additives to the electrolytes .............................................. 6.6.1. Introduction ...................................................... 6.6.2. Effect of additives on the electroendosmotic flow ......................... 6.6.3. Effect of additives on the micro-sensing electrodes ........................ 6.6.4.Additives ........................................................ 6.7. Coating of the micro-sensing electrodes .................................. 6.7.1. Introduction ................................................. 6.7.2. Experimental ............. .................................. 6.8. Detection limits ....................................................... 6.8.1. Introduction ..................................................... 6.8.2.Experimental .................................................... 6.9.Conclusion ........................................................... References ............................................................... 7 . Instrumentation ........................................................... Summary ................................................................ 7.1.Introduction .......................................................... 7.2. Injection systems ...................... ............................. 7.2.1. Introduction ..................................................... 7.2.2.Four-way tap ..................................................... 7.2.3. Six-way valve ............ .................................... 7.2.4. Injection block .................. .............................. 7.2.5. Simplified injection block . . .................................... 7.3. Counter electrode compartments .......................................... 7.3.1. Introduction ..................................................... 7.3.2. Cylindrical counter electrode compartment ............................. 7.3.3. Counter electrode compartment with flat membrane ....................... 7.4.Equipment ........................................................... 7.4.1. Introduction .................................................... 7.4.2. Narrow-bore tube surrounded with a water-jacket ......................... 7.4.3. Narrow-bore tube thermostated with an aluminium block ................... 7.4.4. Equipment with high-resolution detectors ............................... 7.5. Counter flow of electrolyte ............................................... 7.5.1. Introduction ..................................................... 7.5.2. Counter flow with level regulation ..................................... 7.5.3. Counter flow with light-dependent resistor regulation ...................... 7.5.4. Counter flow with direct control on the pumping mechanism via the power supply ..................................................... 7.5.5. Counter flow with no regulation ...................................... 7.5.6. Counter flow regulated by the cur -stabilized power supply; the membrane pump ............. .................................. 135 140 143 143 153 153 155 159 161 164 165 171 171 171 174 180 191 191 191 193 193 196 199 201 203 203 203 203 203 204 205 208 211 211 211 213 215 217 217 219 221 224 230 230 231 233 237 238 24 1
X CONTENTS APPLICATIONS 8.Introduction ............................................................... Summary ................................................................ 8.Introduction ............................................................ 9 . Practical aspects ............................................................ Summary ................................................................ 9.1.Introduction .......................................................... 9.2. Disturbances caused by hydrogen and hydroxyl ions ............................ 9.2.1. Disturbances from the terminator zone in unbuffered systems ............... 9.2.1.1. HI-MI boundary ........................................... 9.2.1.2. MI-MI, boundary ... .................................... 9.2.2. Disturbances from the leading zone in unbuffered systems .................. 9.2.3. Disturbances due to the presence of hydrogen and hydroxyl ions in buffered systems ......................................................... 9.3. Disturbances due to the presence of carbon dioxide ............................ 9.4. Enforced isotachophoresis ............................................... 9.4.1. Disc electrophoresis ................................................ 9.5. Water as terminator ..................................................... 9.6. Purification of the terminator ............................................. 9.7. Conversion of data measured with different detectors ........................... References ............................................................... 10 . Quantitative aspects ........................................................ Summary ............................................................... 10.1.Introduction ......................................................... 10.2.Theoretical .......................................................... 10.3. Thermometric measurements ............................................ 10.3.1. Reproducibility ................................................. 10.3.2. Calibration constant ............................................. 10.4. Conductimetric measurements ........................................... 10.4.1. Reproducibility ................................................. 10.4.2. Calibration constant ............................................. 10.5.Conclusion ........................................................... References ............................................................... 11 . Separation of cationic species in aqueous solutions ................................ Summary ................................................................ 11.1. Separation of cationic species in aqueous solutions using a thermocouple as detector . . 11.1.1. The system WHCl ............................................... 11.1.2. The system WHIO .............................................. 11.1.3. The system WKAC ............................................... 11.1.4. The system WKCAC ............................................. 11.1.5. The system WKDIT .............................................. 11.2. Separation of cationic species in water and deuterium oxide using a conductivity detector (a.c. method) and a UV absorption detector (256 nm) ................... 12 . Separation of anionic species in aqueous solutions ................................. Summary ................................................................. 12.1. Separation of anionic species in aqueous solut'ions using a thermometric detector ..... 12.1.1. Operational system histidine/histidine hydrochloride (pH 6) ............... 12.1.2. Operational system imidazole/imidazole hydrochloride (pH 7) ............. 12.2. Separation of anionic species in aqueous solutions using a conductivity detector (a.c. method) and a UV absorption detector (256 nm) ............................. 12.2.1. Introduction ................................................... 249 249 249 253 253 253 253 253 254 254 257 260 263 264 265 267 268 210 271 273 213 273 214 215 275 215 219 279 280 281 282 283 283 283 285 286 288 289 293 293 295 295 295 295 296 300 300
Page 2 and 3: Journal of Chromatogaphy Library -
Page 4 and 5: Journal of Chromatography Library -
Page 6 and 7: Dedicated to Pr0f.Dr.h. A.I.M. Keul
Page 8 and 9: Contents Preface ..................
Page 12 and 13: CONTENTS XI 12.2.2. Applications ..
Page 14 and 15: It is very well known that charged
Page 16 and 17: Chapter 1 Historical review SUMMARY
Page 18 and 19: HISTORICAL REVIEW 3 Independently i
Page 20 and 21: THEORY
Page 22 and 23: Chapter 2 Principles of electrophor
Page 24 and 25: MOVINGBOUNDARY ELECTROPHORESIS a S
Page 26 and 27: MOVINGBOUNDARY ELECTROPHORESIS 0 iu
Page 28 and 29: ISOTACHOPHORESIS 13 2.4. PRINCIPLE
Page 30 and 31: ISOTACHOPHORESIS 15 As the anionic
Page 32 and 33: ISOTACHOPHORESIS velocities, so tha
Page 34 and 35: ISOTACHOPHORESIS 19 d.t ’.t I Fig
Page 36 and 37: ISOTACHOPHORESIS 21 t Fig.2.8. Isot
Page 38 and 39: ISOELECTRIC FOCUSING 23 The four is
Page 40 and 41: DISCUSSION 25 Fig.2.11. Survey of t
Page 42 and 43: Chapter 3 Concept of mobility SUMMA
Page 44 and 45: IONIC MOBILITY AND IONIC EQUIVALENT
Page 46 and 47: EFFECTIVE IONIC MOBILITY 31 obtain
Page 48 and 49: EFFECTIVE IONIC MOBILITY 33 compari
Page 50 and 51: EFFECTIVE IONIC MOBILITY Fig.3.4. R
Page 52 and 53: DETERMINATION OF IONIC MOBILITIES 3
Page 54 and 55: DETERMINATION OF IONIC MOBILITIES 3
Page 56 and 57: Chapter 4 Mathematical model for is
Page 58 and 59: GENERAL EQUATIONS 43 anionic specie
Page 60 and 61:
GENERAL EQUATIONS 4.2.1. Equilibriu
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GENERAL EQUATIONS 41 i Similar equa
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GENERAL EQUATIONS 49 Uthzone (U-llt
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GENERAL EQUATIONS 4.2.4. Modified O
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GENERAL EQUATIONS TABLE 4.2 REDUCED
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MATHEMATICAL MODEL FOR THE STEADY S
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VALIDITY OF THE ISOTACHOPHORETIC MO
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CHECK OF THE ISOTACHOPHORETIC MODEL
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CHECK OF THE ISOTACHOPHORETIC MODEL
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REFERENCES 81 Fig.4.17. Relationshi
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Chapter 5 Choice of electrolyte sys
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CHOICE OF THE SOLVENT 85 In general
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CHOICE OF THE SOLVENT TABLE 5.2 THE
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CHOICE OF THE SOLVENT pH and pKvalu
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CHOICE OF THE SOLVENT 91 TABLE 5.4
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CHOICE OF THE pH OF THE LEADING ELE
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CHOICE OF THE pH OF THE LEADING ELE
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CHOICE OF THE TERMINATING AND LEADI
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ADDITIONS TO THE ELECTROLYTE SOLUTI
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EXAMPLES Choice of solvent. Can wat
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EXAMPLES 103 solvent, hoping that a
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EXAMPLES TABLE 5.11 STEP HEIGHTS OF
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EXAMPLES TABLE 5.12 STEP HEIGHTS OF
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Fig.5.9. Isotachopherograms of the
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EXAMPLES II 4 3 Fig.5.11. Isotachop
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REFERENCES 113 Example I. As exampl
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INSTRUMENTATION
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Chapter 6 Detection systems SUMMARY
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THERMOMETRIC RECORDING 6.1.3. Combi
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THERMOMETRIC RECORDING Potential gr
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THERMOMETRIC RECORDING /2 Fig.6.1.
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THERMOMETRIC RECORDING T t l Fig.6.
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THERMOMETRIC RECORDING 7 127 It-- I
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THERMOMETRIC RECORDING a concentrat
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HIGH-FREQUENCY CONDUCTIVITY DETECTI
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CONDUCTIVITY DETECTION 133 Fig.6.9.
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CONDUCTIVITY DETECTION 135 The d.c.
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CONDUCTIVITY DETECTION 137 Hi V 1kR
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CONDUCTIVITY DETECTION Fig.6.12. Co
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CONDUCTIVITY DETECTION Fig.6.14. Th
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CONDUCTIVITY DETECTION 143 6.4.4. T
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CONDUCTIVITY DETECTION 145 contact
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CONDUCTIVITY DETECTION 147 We can n
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CONDUCTIVITY DETECTION 149 N.Y., U.
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CONDUCTIVITY DETECTION 151 achieved
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UV ABSORPTION METER 153 -_-.- +- R
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UV ABSORPTION METER Fig.6.22. Contr
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W ABSORPTION METER r +15V -15V Mod
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W ABSORPTION METER 159 TABLE 6.4 PR
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W ABSORPTION METER 230 240 2% m zm
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to Mod (Fig 6.24) oscillator I sync
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UV ABSORPTION METER 165 dicular to
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Fig.6.31. Isotachopherograms for th
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UV ABSORPTION METER 169 50 Fig.6.33
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ADDITIVES TO THE ELECTROLYTES 171 6
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ADDITIVES TO THE ELECTROLYTES 173 e
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ADDITIVES TO THE ELECTROLYTES 175 a
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ADDITIVES TO THE ELECTROLYTES 177 o
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ADDITIVES TO THE ELECTROLYTES 179 d
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ADDITIVES TO THE ELECTROLYTES 181 T
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ADDITIVES TO THE ELECTROLYTES Fig.6
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ADDITIVES TO THE ELECTROLYTES 185 F
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ADDITIVES TO THE ELECTROLYTES 1 2 3
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ADDITIVES TO THE ELECTROLYTES 189 n
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COATING OF THE MICRO-SENSING ELECTR
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DETECTION LIMITS 193 electric drivi
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DETECTION LIMITS 195 Fig.6.48. Infl
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DETECTION LIMITS 197 TABLE 6.6 SURV
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CONCLUSION 199 average length of a
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REFERENCES 201 seen. Again it is cl
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Chapter 7 Instrumentation SUMMARY T
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INJECTION SYSTEMS I! I n Fig.7.1. P
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INJECTION SYSTEMS Fig.7.3. Exploded
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INJECTION SYSTEMS Fig.7.5. Injectio
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COUNTER ELECTRODE COMPARTMENTS 21 1
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COUNTER ELECTRODE COMPARTMENTS Even
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COUNTER ELECTRODE COMPARTMENTS 21 5
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EQUIPMENT the large bore, a more di
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EQUIPMENT 219 belonging to three cl
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EQUIPMENT 221 signal of the thermoc
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EQUIF'MENT 223 No effect on the res
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EQUIPMENT 225 Fig.7.14. Electrophor
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EQUIPMENT Fig.7.16. Photograph of t
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EQUIPMENT 229 mounted equiplanar, w
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COUNTER FLOW OF ELECTROLYTE 231 ins
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COUNTER FLOW OF ELECTROLYTE p High
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COUNTER FLOW OF ELECTROLYTE Fig.7.2
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COUNTER FLOW OF ELECTROLYTE 231 aro
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COUNTER FLOW OF ELECTROLYTE p high
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COUNTER FLOW OF ELECTROLYTE 24 1 7.
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COUNTER FLOW OF ELECTROLYTE I I 31
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COUNTER FLOW OF ELECTROLYTE max. 15
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APPLICATIONS
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Chapter 8 Introduction SUMMARY In t
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INTRODUCTION 25 1 the various ionic
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Chapter 9 Practical aspects SUMMARY
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DISTURBANCES CAUSED BY H+ AND OH 25
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DISTURBANCES CAUSED BY H+ AND OH- 2
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DISTURBANCES DUE TO CO, 263 Electro
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ENFORCED ISOTACHOPHORESIS 265 t T T
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WATER AS TERMINATOR 267 function of
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PURIFICATION OF THE TERMINATOR 7 4
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REFERENCES 271 AS an example, the c
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Chapter 10 Quantitative aspects SUM
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THERMOMETRIC MEASUREMENTS 215 vj c
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THERMOMETRIC MEASUREMENTS TABLE 10.
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CONDUCTIMETRIC MEASUREMENTS factor
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CONCLUSION 28 1 Tables 10.3-10.5 wa
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Chapter 11 Separation of cationic s
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SEPARATION USING A THERMOCOUPLE AS
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I SEPARATION USING A THERMOCOUPLE A
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SEPARATION USING CONDUCTIVITY AND U
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Chapter I2 Separation of anionic sp
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SEPARATION USING A THERMOMETRIC DET
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SEPARATION USING CONDUCTIVITY AND W
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Chapter 13 Amino acids, peptides an
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AMINO ACIDS TABLE 13.1 OPERATIONAL
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AMINO ACIDS 8- ?- 6- 5- 4- 3- 2- f-
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AMINO ACIDS 317 TABLE 13.5 STEP HEI
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AMINO ACIDS Fig.13.3. Schematic dia
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AMINO ACIDS TABLE 13.6 DETERMINATIO
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SEPARATION OF PROTEINS IN AMPHOLYTE
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SEPARATION OF SMALL PEPTIDES reprod
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Chapter I4 Separation of nucleotide
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Chapter 15 Enzymatic reactions SUMM
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ENZYMATIC CONVERSION OF GLUCOSE (FR
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ENZYMATIC CONVERSION OF PYRUVATE 35
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Chapter 16 Separations in non-aqueo
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SEPARATION OF ANIONIC SPECIES IN ME
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SEPARATION OF CATIONIC SPECIES IN M
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EXPERIMENTS IN AQUEOUS METHANOLIC S
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Chapter I7 Counter flow of electrol
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INTRODUCTION 317 In various operati
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EXPERIMENTAL 0% 10 % a b 60 % a b a
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EXPERIMENTAL 381 caused by the coun
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CONCLUSION 383 c t r ’r; I I"
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APPENDICES
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Appendix A Simplified model of movi
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PROCEDURE OF COMPUTATION A.3. PROCE
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EXPERIMENTAL 39 1 TABLE A1 THEORETI
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DISCUSSION E I I Fig.A2. Graphical
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Appendix B Diameter of the narrow-b
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Appendix C Literature 1897-1966 F.
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LITERATURE 399 B.P. Konstantinov an
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LITERATURE 401 1971 L. Arlinger, Is
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LITERATURE 403 1973 L. Arlinger and
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LITERATURE 405 A. Chrambach and J.S
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LITERATURE 407 M. Coxon and M.J. Bi
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Symbols and abbreviations SYMBOLS A
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SYMBOLS AND ABBREVIATIONS SUPERSCRI
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Subject index A Absorption meter, U
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SUBJECT INDEX 41 5 Detection limit
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SUBJECT INDEX 41 7 --_ , separation
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