Frans_M_Everaerts_Isotachophoresis_378342.pdf

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REFERENCES 201 seen. Again it is clear that in order to observe a sample zone with a thermometric detector the zone length must be greater (Table 6.2) than if a high-resolution detector is applied (UV, conductimeter). Hence more sample has been introduced into the system and consequently a longer time of analysis is needed. In Fig.6.51, an analysis under conditions identical with those in Fig.6.50 is shown, recording being effected with a linearized conductimeter (Fig.6.18) and a W-absorption detector (256 nm) (Fig.6.26). It should be pointed out that the various isotachopherograms shown in this chapter are given mainly for pattern recognition, in order to show different effects such as sharpness of the profiles, electrode reactions and impurities. It is difficult to compare one isotachopherogram with another, although a single test mixture of anions was used. In preparing the manuscript, the various isotachopherograms were treated photographically in order to make comparisons simpler. In the Section Applications (Chapters 8-17) the time axis is also given on the relevant isotachopherograms, so that the qualitative and quantitative information can be deduced more easily. Although electrode reactions may sometimes still occur during detection with a conductimeter with the electrodes in direct contact with the electrolyte, its resolution is lugh compared with that of other types of universal detectors. Moreover, possible coating of the electrode can easily be observed by using a combination of an a.c. and d.c. detector (see also Fig.8.1.). We recommend that the entire system should be cleaned from time to time with a non-ionic surfactant, which can be purified by running it through a mixed- bed ion exchanger, because all types of material may be adsorbed on walls made of Perspex, TPX, Pt or even PTFE. When adsorbed, these impurities change the {-potential and hence the electroendosmosis, and thus the resolution of both conductivity and UV absorption detectors is decreased. For effective rinsing of the electrophoretic equipment, we recommend the surfactant Extran (Merck, Darmstadt, G.F.R). REFERENCES 1 F.M. Everaerts, Thesis, University of Technology, Eindhoven, 1968. 2 F.M. Everaerts and Th.P.E.M. Verheggen,J. Chromatogr., 53 (1970) 315. 3 F.M. Everaerts and Th.P.E.M. Verheggen, in P.G. Righetti (Editor), Progress in Isoelectric Focusing and Zsotachophoresis, North-Holland, Amsterdam, and Elsevier, New York, 1975, p. 309. 4 F.M. Everaerts, J.L. Beckers and Th.P.E.M. Verheggen, Ann. NY. Acad. Sci., 209 (1973) 419. 5 F.M. Everaerts, Graduation Rep., University of Technology, Eindhoven, 1964. 6 F.M. Everaerts and Th.P.E.M. Verheggen, J. Chromatog., 91 (1974) 837. 7 F.M. Everaerts and P.J. Rommers,J. Chromatogr., 91 (1974) 809. 8 F.M. Everaerts and Th.P.E.M. Verheggen,J. Chromatogr., 73 (1972) 193. 9 F.M. Everaerts, internal report, University of Technology, Eindhoven, 972. 10 A. Vestermark and B. Sjodin, J. Chromatogr., 71 (1972) 588. 11 L. Arlinger and H. Lundin, Protides Biol. Fluids, Proc. Colloq., 21 (1973) 667. 12 A.J.P. Martin and F.M. Everaerts, Anal. Chim. Ac~Q, 38 (1967) 233. 13 A.J.P. Martin and F.M. Everaerts, Proc. Roy. SOC., Ser. A, 316 (1970) 493. 14 B.P. Konstantinov and O.V. Oshurkova, Sov. Phys.-Tech. Phys., 37 (1967) 1745. 15 I. Vacik, J. Zuska, F.M. Everaerts and Th.P.E.M. Verheggen, Chem. Listy, 66 (1972) 545. 16 F.M. Everaerts, J. Vacik, Th.P.E.M. Verheggen and J. Zuska, J. Chromatogr., 49 (1970) 262. 17 M. Coxon and M.J. Binder,J. Chromatogr., 101 (1974) 1.
202 DETECTION SYSTEMS 18 Publication No. 556 f424, AGA, Infrared Instruments Department, Lidingo, Sweden, 1974. 19 J.L. Fergason, Sci. Amer., 231 (1974) 76. 20 J.L. Beckers, Thesis, University of Technology, Eindhoven, 1973. 21 M. Demjanenko, J. Vacik and .I. Zuska, Chem Listy, in press. 22 Th.P.E.M. Verheggen, E.C. van Ballegooijen, C.H. Massen and F.M. Everaerts, J. Chromatogr., 64 (1972) 185. 23 D.I. Shernoff, Rev. Sci. Instrum, 40 (1969) 1418.
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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.
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 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
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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
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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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