Frans_M_Everaerts_Isotachophoresis_378342.pdf

More documents

Recommendations

Info

AMINO ACIDS TABLE 13.1 OPERATIONAL SYSTEM AT pH 5.4 SUITABLE FOR CATIONIC SEPARATIONS Solvent: H, 0. Electric current @A): Ca. 50-100. Electrolyte -~ Leading Terminating Cation K+ DL-Ala’ Concentration 0.01 N Ca. 0.01 N Counter ion PH CH, COO- 5.4 OH- [added as Ba(OH),] >I Additive 0.05% Polyvinyl alcohol (Mowiol) None nitrogen. Moreover, barium hydroxide of pro analysi quality must be added to the terminating electrolyte in order to prevent any carbonate (hydrogen carbonate) from penetrating into the narrow-bore tube via the reservoir filed with the terminating electrolyte. The addition of barium hydroxide to the leading electrolyte did not improve the separation, however. Because of the high mobility of Ba2+, less sharp boundaries can be expected. If suitable precautions are taken, some carbonate (hydrogen carbonate) may still be detected if an anion with a lugher effective mobility is used as the leading ion. This carbonate (hydrogen carbonate), however, did not obscure both the qualitative and quantitative results. Only a leading ion with an effective mobility higher than or equal to that of the carbonate (hydrogen carbonate) ion can be used, otherwise the carbonate may no longer be visible but may still disturb or at least obscure the analytical result, if it is supported continuously. The resolution will decrease and zone electrophoretic phenomena can soon be expected e.g., ‘elution’ by the carbonate (hydrogen carbonate) ions. Several suitable buffers are commercially available that enable one to work at high pH. Some of them, including bis(3-aminopropyl)amine, trime thylenediamine, L-arginine, 1-methylpiperidine, octylamine, ethanolamine and L-lysine, were tested for purity and effective mobility*. Some of the compounds were found to be very impure and even could not be purified satisfactorily by the usual methods such as distillation, recrystallization or ion exchange. Some of the counter ions show unexpected phenomena, e.g., enforced isotachophoretic systems or undesirable complex formation. Of the compounds mentioned above, only 1 -methyl piperidine, L-arginine, L-lysine, octylamine and ethanolamine gave good results. In Tables 13.2 and 13.3, two operational systems are given in which analyses were performed and for which more data will be given later. *If a counter ion (buffer) is too mobile, a considerable proportion of the electricity is carried by this counter ion, which results in less sharp zone boundaries and a decrease in resolution. 31 3
314 TABLE 13.2 AMINO ACIDS, PEFTIDES AND PROTEINS OPERATIONAL SYSTEM AT pH ABOUT 9 SUITABLE FOR ANIONIC SEPARATIONS Solvent: H, 0. Electric current (/.LA): CQ. 50-100. Electrolyte Leading Terminating __- Anion 5-Br-2,4-diOH-C6 H, COO- 0-Ala- (recrystallized from waterethanol) Concentration 0.004 M CQ. 0.01 M Counter ion HOC, H, N'H Baa+ [added as Ba(OH),] PH 9.0, 9.2, 9.4, 9.6 Ca. 10.5 Additive 0.05% Polyvinyl None alcohol (Mowiol) TABLE 13.3 OPERATIONAL SYSTEM AT pH ABOUT 9 SUITABLE FOR ANIONIC SEPARATIONS Solvent: H, 0. Electric current (fiA): Ca. 50-100. Electrolyte Leading Terminating Anion 5-Br-2,4-diOH-C6 H, COO- P- Ala- (recrystallized from wa tere thanol) Concentration 0.004 M Ca. 0.01 M Counter ion L-Lys+ Ba2+ [added as Ba(OH), J PH 9.1, 9.2, 9.4 Ca. 10.5 Additive 0.05% Polyvinyl alcohol (Mowiol) None The leading ion in these systems was chosen because its effective mobility is almost identical with the effective mobility of the carbonate (hydrogen carbonate) ion. In the linear trace of the conductivity detector, the carbonate (hydrogen carbonate) step is no longer visible*. The leading ion, however, has a W absorption. If too much carbonate (hydrogen carbonate) is present, it is present in the UV trace, which is used only to mark the UV-absorbing zones and has so far not been applied for qualitative determinations. The carbonate (hydrogen carbonate), if present, is enriched just before the zones of the isotachophoretic 'train', as can be seen in Fig. 13.1. *This simplifies the qualitative information considerably.
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 10 and 11:
CONTENTS IX 6.4.2. The d.c. method
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
Page 62 and 63:
GENERAL EQUATIONS 41 i Similar equa
Page 64 and 65:
GENERAL EQUATIONS 49 Uthzone (U-llt
Page 66 and 67:
GENERAL EQUATIONS 4.2.4. Modified O
Page 68 and 69:
GENERAL EQUATIONS TABLE 4.2 REDUCED
Page 70 and 71:
MATHEMATICAL MODEL FOR THE STEADY S
Page 72 and 73:
Page 74 and 75:
Page 76 and 77:
Page 78 and 79:
Page 80 and 81:
Page 82 and 83:
Page 84 and 85:
VALIDITY OF THE ISOTACHOPHORETIC MO
Page 86 and 87:
Page 88 and 89:
Page 90 and 91:
Page 92 and 93:
CHECK OF THE ISOTACHOPHORETIC MODEL
Page 94 and 95:
CHECK OF THE ISOTACHOPHORETIC MODEL
Page 96 and 97:
REFERENCES 81 Fig.4.17. Relationshi
Page 98 and 99:
Chapter 5 Choice of electrolyte sys
Page 100 and 101:
CHOICE OF THE SOLVENT 85 In general
Page 102 and 103:
CHOICE OF THE SOLVENT TABLE 5.2 THE
Page 104 and 105:
CHOICE OF THE SOLVENT pH and pKvalu
Page 106 and 107:
CHOICE OF THE SOLVENT 91 TABLE 5.4
Page 108 and 109:
CHOICE OF THE pH OF THE LEADING ELE
Page 110 and 111:
CHOICE OF THE pH OF THE LEADING ELE
Page 112 and 113:
CHOICE OF THE TERMINATING AND LEADI
Page 114 and 115:
ADDITIONS TO THE ELECTROLYTE SOLUTI
Page 116 and 117:
EXAMPLES Choice of solvent. Can wat
Page 118 and 119:
EXAMPLES 103 solvent, hoping that a
Page 120 and 121:
EXAMPLES TABLE 5.11 STEP HEIGHTS OF
Page 122 and 123:
EXAMPLES TABLE 5.12 STEP HEIGHTS OF
Page 124 and 125:
Fig.5.9. Isotachopherograms of the
Page 126 and 127:
EXAMPLES II 4 3 Fig.5.11. Isotachop
Page 128 and 129:
REFERENCES 113 Example I. As exampl
Page 130 and 131:
INSTRUMENTATION
Page 132 and 133:
Chapter 6 Detection systems SUMMARY
Page 134 and 135:
THERMOMETRIC RECORDING 6.1.3. Combi
Page 136 and 137:
THERMOMETRIC RECORDING Potential gr
Page 138 and 139:
THERMOMETRIC RECORDING /2 Fig.6.1.
Page 140 and 141:
THERMOMETRIC RECORDING T t l Fig.6.
Page 142 and 143:
THERMOMETRIC RECORDING 7 127 It-- I
Page 144 and 145:
THERMOMETRIC RECORDING a concentrat
Page 146 and 147:
HIGH-FREQUENCY CONDUCTIVITY DETECTI
Page 148 and 149:
CONDUCTIVITY DETECTION 133 Fig.6.9.
Page 150 and 151:
CONDUCTIVITY DETECTION 135 The d.c.
Page 152 and 153:
CONDUCTIVITY DETECTION 137 Hi V 1kR
Page 154 and 155:
CONDUCTIVITY DETECTION Fig.6.12. Co
Page 156 and 157:
CONDUCTIVITY DETECTION Fig.6.14. Th
Page 158 and 159:
CONDUCTIVITY DETECTION 143 6.4.4. T
Page 160 and 161:
CONDUCTIVITY DETECTION 145 contact
Page 162 and 163:
CONDUCTIVITY DETECTION 147 We can n
Page 164 and 165:
CONDUCTIVITY DETECTION 149 N.Y., U.
Page 166 and 167:
CONDUCTIVITY DETECTION 151 achieved
Page 168 and 169:
UV ABSORPTION METER 153 -_-.- +- R
Page 170 and 171:
UV ABSORPTION METER Fig.6.22. Contr
Page 172 and 173:
W ABSORPTION METER r +15V -15V Mod
Page 174 and 175:
W ABSORPTION METER 159 TABLE 6.4 PR
Page 176 and 177:
W ABSORPTION METER 230 240 2% m zm
Page 178 and 179:
to Mod (Fig 6.24) oscillator I sync
Page 180 and 181:
UV ABSORPTION METER 165 dicular to
Page 182 and 183:
Fig.6.31. Isotachopherograms for th
Page 184 and 185:
UV ABSORPTION METER 169 50 Fig.6.33
Page 186 and 187:
ADDITIVES TO THE ELECTROLYTES 171 6
Page 188 and 189:
ADDITIVES TO THE ELECTROLYTES 173 e
Page 190 and 191:
ADDITIVES TO THE ELECTROLYTES 175 a
Page 192 and 193:
ADDITIVES TO THE ELECTROLYTES 177 o
Page 194 and 195:
ADDITIVES TO THE ELECTROLYTES 179 d
Page 196 and 197:
ADDITIVES TO THE ELECTROLYTES 181 T
Page 198 and 199:
ADDITIVES TO THE ELECTROLYTES Fig.6
Page 200 and 201:
ADDITIVES TO THE ELECTROLYTES 185 F
Page 202 and 203:
ADDITIVES TO THE ELECTROLYTES 1 2 3
Page 204 and 205:
ADDITIVES TO THE ELECTROLYTES 189 n
Page 206 and 207:
COATING OF THE MICRO-SENSING ELECTR
Page 208 and 209:
DETECTION LIMITS 193 electric drivi
Page 210 and 211:
DETECTION LIMITS 195 Fig.6.48. Infl
Page 212 and 213:
DETECTION LIMITS 197 TABLE 6.6 SURV
Page 214 and 215:
CONCLUSION 199 average length of a
Page 216 and 217:
REFERENCES 201 seen. Again it is cl
Page 218 and 219:
Chapter 7 Instrumentation SUMMARY T
Page 220 and 221:
INJECTION SYSTEMS I! I n Fig.7.1. P
Page 222 and 223:
INJECTION SYSTEMS Fig.7.3. Exploded
Page 224 and 225:
INJECTION SYSTEMS Fig.7.5. Injectio
Page 226 and 227:
COUNTER ELECTRODE COMPARTMENTS 21 1
Page 228 and 229:
COUNTER ELECTRODE COMPARTMENTS Even
Page 230 and 231:
COUNTER ELECTRODE COMPARTMENTS 21 5
Page 232 and 233:
EQUIPMENT the large bore, a more di
Page 234 and 235:
EQUIPMENT 219 belonging to three cl
Page 236 and 237:
EQUIPMENT 221 signal of the thermoc
Page 238 and 239:
EQUIF'MENT 223 No effect on the res
Page 240 and 241:
EQUIPMENT 225 Fig.7.14. Electrophor
Page 242 and 243:
EQUIPMENT Fig.7.16. Photograph of t
Page 244 and 245:
EQUIPMENT 229 mounted equiplanar, w
Page 246 and 247:
COUNTER FLOW OF ELECTROLYTE 231 ins
Page 248 and 249:
COUNTER FLOW OF ELECTROLYTE p High
Page 250 and 251:
COUNTER FLOW OF ELECTROLYTE Fig.7.2
Page 252 and 253:
COUNTER FLOW OF ELECTROLYTE 231 aro
Page 254 and 255:
COUNTER FLOW OF ELECTROLYTE p high
Page 256 and 257:
COUNTER FLOW OF ELECTROLYTE 24 1 7.
Page 258 and 259:
COUNTER FLOW OF ELECTROLYTE I I 31
Page 260 and 261:
COUNTER FLOW OF ELECTROLYTE max. 15
Page 262 and 263:
APPLICATIONS
Page 264 and 265:
Chapter 8 Introduction SUMMARY In t
Page 266 and 267:
INTRODUCTION 25 1 the various ionic
Page 268 and 269:
Chapter 9 Practical aspects SUMMARY
Page 270 and 271:
DISTURBANCES CAUSED BY H+ AND OH 25
Page 272 and 273:
Page 274 and 275:
DISTURBANCES CAUSED BY H+ AND OH- 2
Page 276 and 277:
Page 278 and 279: DISTURBANCES DUE TO CO, 263 Electro
Page 280 and 281: ENFORCED ISOTACHOPHORESIS 265 t T T
Page 282 and 283: WATER AS TERMINATOR 267 function of
Page 284 and 285: PURIFICATION OF THE TERMINATOR 7 4
Page 286 and 287: REFERENCES 271 AS an example, the c
Page 288 and 289: Chapter 10 Quantitative aspects SUM
Page 290 and 291: THERMOMETRIC MEASUREMENTS 215 vj c
Page 292 and 293: THERMOMETRIC MEASUREMENTS TABLE 10.
Page 294 and 295: CONDUCTIMETRIC MEASUREMENTS factor
Page 296 and 297: CONCLUSION 28 1 Tables 10.3-10.5 wa
Page 298 and 299: Chapter 11 Separation of cationic s
Page 300 and 301: SEPARATION USING A THERMOCOUPLE AS
Page 306 and 307: I SEPARATION USING A THERMOCOUPLE A
Page 308 and 309: SEPARATION USING CONDUCTIVITY AND U
Page 310 and 311: Chapter I2 Separation of anionic sp
Page 312 and 313: SEPARATION USING A THERMOMETRIC DET
Page 316 and 317: SEPARATION USING CONDUCTIVITY AND W
Page 326 and 327: Chapter 13 Amino acids, peptides an
Page 330 and 331: AMINO ACIDS 8- ?- 6- 5- 4- 3- 2- f-
Page 332 and 333: AMINO ACIDS 317 TABLE 13.5 STEP HEI
Page 334 and 335: AMINO ACIDS Fig.13.3. Schematic dia
Page 336 and 337: AMINO ACIDS TABLE 13.6 DETERMINATIO
Page 338 and 339: SEPARATION OF PROTEINS IN AMPHOLYTE
Page 350 and 351: SEPARATION OF SMALL PEPTIDES reprod
Page 352 and 353: Chapter I4 Separation of nucleotide
Page 362 and 363: Chapter 15 Enzymatic reactions SUMM
Page 364 and 365: ENZYMATIC CONVERSION OF GLUCOSE (FR
Page 370 and 371: ENZYMATIC CONVERSION OF PYRUVATE 35
Page 376 and 377: Chapter 16 Separations in non-aqueo
Page 378 and 379:
SEPARATION OF ANIONIC SPECIES IN ME
Page 380 and 381:
SEPARATION OF CATIONIC SPECIES IN M
Page 382 and 383:
Page 384 and 385:
Page 386 and 387:
Page 388 and 389:
EXPERIMENTS IN AQUEOUS METHANOLIC S
Page 390 and 391:
Chapter I7 Counter flow of electrol
Page 392 and 393:
INTRODUCTION 317 In various operati
Page 394 and 395:
EXPERIMENTAL 0% 10 % a b 60 % a b a
Page 396 and 397:
EXPERIMENTAL 381 caused by the coun
Page 398 and 399:
CONCLUSION 383 c t r ’r; I I"
Page 400 and 401:
APPENDICES
Page 402 and 403:
Appendix A Simplified model of movi
Page 404 and 405:
PROCEDURE OF COMPUTATION A.3. PROCE
Page 406 and 407:
EXPERIMENTAL 39 1 TABLE A1 THEORETI
Page 408 and 409:
DISCUSSION E I I Fig.A2. Graphical
Page 410 and 411:
Appendix B Diameter of the narrow-b
Page 412 and 413:
Appendix C Literature 1897-1966 F.
Page 414 and 415:
LITERATURE 399 B.P. Konstantinov an
Page 416 and 417:
LITERATURE 401 1971 L. Arlinger, Is
Page 418 and 419:
LITERATURE 403 1973 L. Arlinger and
Page 420 and 421:
LITERATURE 405 A. Chrambach and J.S
Page 422 and 423:
LITERATURE 407 M. Coxon and M.J. Bi
Page 424 and 425:
Symbols and abbreviations SYMBOLS A
Page 426 and 427:
SYMBOLS AND ABBREVIATIONS SUPERSCRI
Page 428 and 429:
Subject index A Absorption meter, U
Page 430 and 431:
SUBJECT INDEX 41 5 Detection limit
Page 432 and 433:
SUBJECT INDEX 41 7 --_ , separation
show all

Frans_M_Everaerts_Isotachophoresis_378342.pdf

Create successful ePaper yourself

Delete template?

Save as template?