Frans_M_Everaerts_Isotachophoresis_378342.pdf

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COUNTER ELECTRODE COMPARTMENTS 21 1 section 7.2.3. The injection block was made of Perspex and PTX. The conical tip of tap (4) was made of silicone rubber (shape 90"). This tap is not available commercially as a single piece. The other taps used were commercially available PTFE-lined Hamilton (1MM1) taps (Hamilton, Bonaduz, Switzerland). 7.2.5. Simplified injection block Because the construction of the injection block described in section 7.2.4 is rather complicated, an injection block of much simpler construction (T-way) is shown in Fig.7.7. Such injection blocks have been made of Perspex, TPX, Kel-F, PTFE and polypropylene. The connection with the narrow-bore tube is similar to that described in section 7.2.3. A Hamilton (1MM 1) PTFE-lined valve is mounted between the injection block and the reservoir containing the terminating electrolyte. Another tap, also a PTFE-lined Hamilton (IMM1) valve, is mounted between the injection block and the drain. The equipment can be rinsed and re-filled with leading electrolyte via a tap at the side of the counter electrode compartment (not shown in Fig.7.7, but shown in Fig.7.16). Tap B is opened during this procedure, while tap A is closed. The tap at the side of the counter electrode compartment is then closed and tap A is opened, while tap B remains open. The terminating electrolyte now flows towards drain. No suction or pump need be applied. After this procedure, tap B is closed and the sample can be introduced via the septum with a standard micro-syringe. In this method of sample introduction, the injection can be made only in the leading electrolyte. A large amount of mobile ions of the leading electrolyte are behind the sample introduced and these ions must overtake the sample ions during the isotachophoretic separation procedure. This may be a complication, especially if the concentration of the sample ions is hgh. Because some of the leading electrolyte is mixed with the terminating electrolyte before the analysis, as a result of the introduction of the needle of the micro-syringe, then after the injection of the sample has been made, tap B must be opened in order to remove the leading electrolyte that is mixed with the terminating electrolyte in the horizontal canal. Because of its simple construction, this type of injection block can be recommended in many instances, especially when some precautions can easily be taken with respect to the leading and terminating electrolytes. The concentration of all of the sample ions must not be too high. 7.3. COUNTER ELECTRODE COMPARTMENTS 7.3.1. Introduction Because in narrow-bore tubing a stabilizing effect is obtained, in most experiments no stabilizing agents (e.g., agar agar, polyacrylamide, arrowroot, dextran, agarose) are added to the electrolytes. The counter flow compartment must therefore consist of a semi- permeable membrane in order to prevent any hydrodynamic flow of electrolyte between the two electrode compartments owing to the difference in levels in these compartments.
212 INSTRUMENTATION Fig.7.7. Simpler injection block for electrophoretic equipment suitable for isotachophoretic analyses. Two Hamilton (1MM1) PTFE-lined valves are applied: between the reservoir of the terminating electrolyte and the I-mm narrow-bore tube (A), and between the 1-mm narrow-bore tube and the drain (B). The sample can be introduced into the narrow-bore tube filled with leading electrolyte, so that it is always mixed with the leading electrolyte. A considerable amount of the leading ion must be behind the sample if no intern standard is applied.
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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
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 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: DISTURBANCES CAUSED BY H+ AND OH 25
Page 274 and 275: DISTURBANCES CAUSED BY H+ AND OH- 2
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DISTURBANCES CAUSED BY H+ AND OH 26
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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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