Frans_M_Everaerts_Isotachophoresis_378342.pdf

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EQUIPMENT 219 belonging to three clearly distinguishable types in the development of the instrument& tion of analytical isotachophoretic equipment with a narrow-bore tube. 7.4.2. Narrow-bore tube surrounded with a water-jacket Fig.7.11 shows a photograph of the isotachophoretic equipment with which experiments were carried out in the early days of isotachophoresis (1964). Instead of the PTFE narrow-bore tube, as in Fig.7.11, a narrow-bore tube made of Pyrex glass was used. The equipment consists of a narrow-bore tube, which is fixed with adhesive (shellac if a Pyrex or Araldite if a PTFE narrow-bore tube is used) in a type of Liebig condenser. On the left-hand side a four-way tap is mounted, as discussed in section 7.2.2. The electrode compartment, which contains the semipermeable membrane as discussed in section 7.3.2, is not mounted as a counter electrode compartment as is usually done, but is mounted behind the sample tap and contains the terminating electrolyte. The reason for this is the fact that we still use this equipment for measurements of mobilities by the moving-boundary method and it is easier to rinse if the membrane compartment is mounted at the position shown in Fig.7.11. By means of the cylindrical membrane electrode compartment, hydrodynamic flow of electrolyte is prevented. The connection between the reservoir containing the terminating electrolyte and the cylindrical membrane electrode compartment is made with a PTFE tube and a PTFE-lined Hamilton (lMF1) valve in which the syringe fits. In the Liebig-type condenser, there are three holes into which the thermocouples (linear and differential) may be mounted. Because these thermocouples are so thin, the narrow-bore tube is fured by a small spring, which fits the tube and which is fixed to the wall of the Liebieg-type condenser with hot shellac. The thermocouples are finally soldered on copper wires, also fixed to the wall of the condenser with hot shellac. In Fig.7.11, three thermocouples, all of the linear type, are mounted. In order to reduce the influence of temperature differences in the laboratory (due to movement, draughts etc.), the holes in the wall of the condenser are sealed with adhesive foil that is covered with aluminium foil. For optimal results, the whole equipment is covered with a blanket of cotton-wool. Thermostated water is circulated through the outer space of the condenser to give a constant temperature inside. The reference junction of the thermocouple is therefore mounted with some adhesive on the inside of the condenser wall, protected with a heat-sink compound. For thermostating in our work, a Hacke thermostat with an accuracy of +O.l"C is used. However, if the variations in the temperature of the laboratory are not too large, no thermostat is needed and the outer space of the condenser can simply be filled with water. The capacity of the outer space and hence the volume of water are large enough to keep the temperature constant in the space where the thermocouples are mounted during the isotachophoretic run. The procedure for filing and cleaning this equipment was described in section 7.2.2. The total length of the narrow-bore tube of the equipment shown in Fig.7.11 is approximately 1 m, while the thermocbuples are mounted at distances of 25, 50 and 75 cm. In our work, the signals derived from the thermocouples are amplified with a hick amplifier (type A) and recorded with a potentiometric recorder. If only the linear
Fig.7.11. Electrophoretic equipment suitable for isotachophoretic analyses, constructed in 1964 by <strong>Everaerts</strong>. Detection is effected with thermocouples (copper-constantan), wound aound the narrow-bore tube at three different positions and fixed with adhesive. I?le narrow-bore tube is mounted in a type of Liebig condenser, filled with thermostated water. The injection can be made via a four-way tap (Fig.7.1.). On the left-hand side is mounted the cylindrical electrode compartment with the cellulose polyacetate semipermeable membrane (Fig.7.8.), which was added at a later stage to the equipment.
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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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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
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Page 232 and 233: EQUIPMENT the large bore, a more di
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 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
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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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