Frans_M_Everaerts_Isotachophoresis_378342.pdf

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SEPARATION USING CONDUCTIVITY AND UV DETECTORS 309 TABLE 12.8 RELATIVE STEP HEIGHTS OBTAINED IN THE OPERATIONAL SYSTEMS AT pH 6 (TABLE 12.1) FOR VARIOUS ANIONIC SPECIES WITH WATER AND DEUTERIUM-OXIDE AS SOLVENTS The accuracy is better than 4%. The values are given for comparison of the two solvents. The experiments with water were carried out at 70 PA and those with deuterium oxide at 80 PA*. In both solvents the chloride ion (leading ion) has a relative step height of 0, while the chlorate ion has a relative step height of 100. h = step height. -- = No UV absorbance. lonic species Aspartic acid, dl-benzyl Acetic acid Acetic acid, monochloro Acetic acid, dichloro Acetic acid, trichloro Benzoic acid Benzoic acid, p-amino Benzoic acid, 2,4-dihydroxy Benzoic acid, p-nitro Butyric acid Cacodylic acid Capric acid Caproic acid Caprylic acid Chlorate Chromic acid Chromic acid, bi Citric acid Enanthic acid Formic acid Furnaric acid Glucuronic acid Glutamic acid Glycolic acid Hippuric acid Iodic acid a-Ketoglutaric acid Lactic acid Laewlinic acid Maleic acid Malic acid Malonic acid Methacrylic acid Naphthalene-2-sulphonic acid Nicotinic acid Nitric acid Nitrous acid Orotic acid 100 h hchlorate 1150 4 84 487 542 620 7 26 836 803 771 751 1509 1486 905 1062 100 134 132 259 988 208 217 1033 927 456 962 464 259 609 744 305 259 209 619 798 731 35 25 7 38 uv absorption (%) 40 100 h hchlorate 1244 519 526 583 669 7 86 923 873 840 818 1787 1814 998 1170 100 146 145 277 1099 217 225 1143 1015 503 1060 486 268 643 801 323 27 2 224 680 858 817 28 17 808 uv absorption (%I (Continued on p. 310)
310 SEPARATION OF ANIONIC SPECIES IN AQUEOUS SOLUTIONS TABLE 12.8 (continued) Ionic species Oxalic acid Pelargonic acid Phenylacetic acid Phosphoric acid Phthalic acid Picric acid Pivalic acid Propionic acid Propionic acid, p-chloro Pyrazine, 2,3-dicarboxylic acid Pyrazole, 3,5-dicarboxylic acid Salicylic acid Succinic acid Sulphamic acid Sulphanilic acid Sulphuric acid Sulphurous acid Tartronic acid HZ0 D*O - - lOOh uv lOOh W hcMorate absorption (%) &hlorate absorption (%) 93 1150 823 614 382 766 859 635 6 29 296 293 647 301 294 688 60 284 157 88 1252 915 677 410 819 972 699 701 321 316 703 314 313 736 57 319 167 *The step height is not influenced by this difference in electric current, if conductivity detection is applied (a.c. method). TABLE 12.9 DIFFERENCES IN THE EFFECTIVE MOBILITIES OF SOME ANIONS IN THE OPERATIONAL SYSTEM AT pH 6 (TABLE 12.1) WITH WATER AND DEUTERIUM OXIDE AS THE SOLVENTS The values (h) are the step heights (mm) that can be measured in the linear trace of the conductivity detector. Ionic species 'H,O hD,O Ionic species hH,O hD,O Sulphate 5.6 5.6 p-chloropropionate 58.4 69.5 Chlorate 9.3 9.8 Benzoate 67.5 76.6 Chromate 12.4 14.2 Naphthalene-2sulphonate 74.0 83.7 Malonate 18.8 21.8 Glutamate 86.1 99.0 Pyrazole-3,5-dicarboxylate 27.5 30.8 Enanthat e 92.2 102.2 Adipate 37.2 42.8 Benzyl-dl-aspartate 107.4 121.6 Acetate 44.9 51.4 ~~
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Journal of Chromatogaphy Library -
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Journal of Chromatography Library -
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Dedicated to Pr0f.Dr.h. A.I.M. Keul
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Contents Preface ..................
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CONTENTS IX 6.4.2. The d.c. method
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CONTENTS XI 12.2.2. Applications ..
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It is very well known that charged
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Chapter 1 Historical review SUMMARY
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HISTORICAL REVIEW 3 Independently i
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THEORY
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Chapter 2 Principles of electrophor
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MOVINGBOUNDARY ELECTROPHORESIS a S
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MOVINGBOUNDARY ELECTROPHORESIS 0 iu
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ISOTACHOPHORESIS 13 2.4. PRINCIPLE
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ISOTACHOPHORESIS 15 As the anionic
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ISOTACHOPHORESIS velocities, so tha
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ISOTACHOPHORESIS 19 d.t ’.t I Fig
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ISOTACHOPHORESIS 21 t Fig.2.8. Isot
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ISOELECTRIC FOCUSING 23 The four is
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DISCUSSION 25 Fig.2.11. Survey of t
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Chapter 3 Concept of mobility SUMMA
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IONIC MOBILITY AND IONIC EQUIVALENT
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EFFECTIVE IONIC MOBILITY 31 obtain
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EFFECTIVE IONIC MOBILITY 33 compari
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EFFECTIVE IONIC MOBILITY Fig.3.4. R
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DETERMINATION OF IONIC MOBILITIES 3
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DETERMINATION OF IONIC MOBILITIES 3
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Chapter 4 Mathematical model for is
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GENERAL EQUATIONS 43 anionic specie
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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 25
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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 328 and 329: AMINO ACIDS TABLE 13.1 OPERATIONAL
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
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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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