Frans_M_Everaerts_Isotachophoresis_378342.pdf

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SEPARATION USING A THERMOMETRIC DETECTOR 299 Fig.12.2. Isotachopherogram of the separation of some anions in the operational system listed in Table 12.1 (pH 6). 1 = Chloride; 2 = sulphate; 3 = chlorate;4 = chromate; 5 = malonate; 6 = pyrazole- 3,5-dicarboxylate; 7 = adipate; 8 = acetate; 9 = p-chloropropionate; 10 = phenylacetate; 11 = ascorbate. The current was stabilized at 70 PA. T= Increasing temperature; t = time. This isotachopherogram is used in several places in this book for comparison of the various detectors and the various solvents. The smaller steps in the system imidazole/imidazole hydrochloride correlate correctly with the calculated zone resistances (see section 4.5). Some step heights are low, which can be ascribed to a dissociation that is more complete in this system. Some compounds that show these very large shifts are citric acid (pK3 = 6.4), orthophosphoric acid (pK, = 7.21) and chromic acid (pKz = 6.49). Fig.12.3 shows aa isotachopherogram of the separation of some anions carried out in the operational system listed in Table 12.2. Although the resolution is lower than in the operational system at pH 6, in general anions can be separated if the step heights given in Table 12.3 differ by about 10%. The disadvantage of this operational system is the relatively high pH, which soon gives a disturbance due to carbonate.
300 I1 rc-- t \ t SEPARATION OF ANIONIC SPECIES IN AQUEOUS SOLUTIONS n Fig.12.3. Isotachopherogram of the separation of some anions carried out in the operational system listed in Table 12.2 @H 7). 1 = Chloride; 2 = sulphate; 3 = oxalate; 4 = chlorate; 5 = formate; 6 = pyrazole-3,5-dicarboxylate; 7 = adipate; 8 = iodate; 9 = P-chloropropionate; 10 =nicotinate; 11 = ascorbate. The current was stabilized at 70 PA. If this isotachopherogram is compared with those in Figs.12.1 and 12.2, it can be seen that the resolution is smaller, although separations are still possible. 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 In this section, we consider isotachophoretic separations in four operational systems with water and deuterium oxide. The pH values were chosen arbitrarily as 7.5 (Table 12.4), 6 (Table 12.1), 4.5 (Table 12.5) and 3 (Table 12.6). The results obtained in these systems are listed in Table 12.7 for water and Table 12.8 for deuterium oxide. The analyses discussed in section 12.2.2 were carried out mainly in these operational systems, although the pH, for optimal separation, may be chosen to be higher or lower than those given in the tables. The analyses were carried out with the equipment described in section 7.4.4.
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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
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 274 and 275: DISTURBANCES CAUSED BY H+ AND OH- 2
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
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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
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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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