Frans_M_Everaerts_Isotachophoresis_378342.pdf

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THERMOMETRIC RECORDING Potential gradient Temperature PH Determined by the direct driving current chosen and the conductivity of the L.E. Determined by the direct driving current chosen and the concentration of the anion of the L.E. (anion separation) or the cation of the L.E. (cation separation) and the effective mobility of the ionic constituent present. Determined by the direct driving current chosen and the concentration of the L.E. initially chosen and the effective mobility of the terminating ion. Determined by the direct driving Determined by the ratio of the current and the conductivity of the L.E. 121 concentration of the ionic species of the leading electrolyte and their pKa values. Determined by the direct driving Determined by the ratio of the current and the concentration concentration of the ionic of the anion of the L.E. (anion species present in each zone separation) or cation of the and their pKa values. L.E. (cation separation). Determined by the direct driving Determined by the ratio of the current and the concen- concentration of the ionic tration of the L.E. initially species present in this zone chosen. and their pKa values. The potential gradient is a constant The temperature is constant for The pH tends to increase in for each zone and increases each zone and increases from anion separation and to from the L.E. towards the T.E. the L.E. towards the T.E. decrease in cation separation (except for ‘enforced’ isotacho- (except for ‘enforced’ iso- from the L.E. towards the phoretic systems). tachophoretic systems). T.E. If this thermocouple is mounted around the narrow-bore tube, a linear stepwise recording of the isotachophoretic zones is obtained as soon as they have passed this fixed thermocouple. In order to make the zone boundaries more pronounced electronically, the differential of this signal can be obtained. An example of a possible circuit is given in Fig.6.2. Another possibility is to make a differential thermocouple with two copper-constantan junctions, about 2 mm apart. This thermocouple can be mounted around the narrow-bore tube in a similar way by means of a suitable elastic adhesive. The two twisted (and silver soldered) ends must be bent until they are approximately at right-angles to the surface of the narrow-bore tube. While the values from the passage of a zone boundary recorded by a normal thermocouple can be expected to be of the order of millivolts, a differential thermocouple indicates the passage of a zone boundary with a signal of the order of microvolts, and amplification is therefore necessary. The differential thermocouple has to be balanced. Normally, there is a difference in temperature between the two junctions of the differential thermocouple, because both
TABLE 6.2 SURVEY OF THE DETECTORS USED IN ANALYTICAL ISOTACHOPHORESIS Lmin. = minimum detectable zone length; Qmin, = minimum detectable amount of ionic component in gramequivalents; t,, = average time for analysis; I = direct driving current; Scan = possrbdity of scanning. Type Performance ‘min.(mm) Qmin. t,,(min) Scan Dependence on 1 General information Thermal Thermocouple, 25 fim 5 (Cu-constantan) thermistor (Philips micro-beat) Conductivity Micro-sensing electrodes 2 (high frequency) not in contact with electrolyte Conductivity Micro-sensing electrodes in contact with electrolyte 0.05 Potential gradient Micro-sensing electrodes 0.05 in contact with electrolyte uv Micro-cell wavelengths: 0.05 205,256,280 and 340 nrn 0.5*109 30 No I= 2.10 0 20 Yes(?) No 0.5 * lo-” 10 No No 0.5 * lo-’ 10 No I Qualitative Quantitative General detector Qualitative Quantitative General detector Qualitative Quantitative General detector* Qualitative Quantitative General detector 0.5 lo-” 10 Yes No Quantitative Specific detector** *A coating on the sensing electrodes gives the detector ‘specific’ characteristics. **The use of strong Wabsorbing counter ions of which the W absorption is a function of the pH changes the specific characteristics of this detector into ‘general’ characteristics. 8 i3 2: cn - N
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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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Page 92 and 93: CHECK OF THE ISOTACHOPHORETIC MODEL
Page 94 and 95: CHECK OF THE ISOTACHOPHORETIC MODEL
Page 96 and 97: REFERENCES 81 Fig.4.17. Relationshi
Page 98 and 99: Chapter 5 Choice of electrolyte sys
Page 100 and 101: CHOICE OF THE SOLVENT 85 In general
Page 102 and 103: CHOICE OF THE SOLVENT TABLE 5.2 THE
Page 104 and 105: CHOICE OF THE SOLVENT pH and pKvalu
Page 106 and 107: CHOICE OF THE SOLVENT 91 TABLE 5.4
Page 108 and 109: CHOICE OF THE pH OF THE LEADING ELE
Page 110 and 111: CHOICE OF THE pH OF THE LEADING ELE
Page 112 and 113: CHOICE OF THE TERMINATING AND LEADI
Page 114 and 115: ADDITIONS TO THE ELECTROLYTE SOLUTI
Page 116 and 117: EXAMPLES Choice of solvent. Can wat
Page 118 and 119: EXAMPLES 103 solvent, hoping that a
Page 120 and 121: EXAMPLES TABLE 5.11 STEP HEIGHTS OF
Page 122 and 123: EXAMPLES TABLE 5.12 STEP HEIGHTS OF
Page 124 and 125: Fig.5.9. Isotachopherograms of the
Page 126 and 127: EXAMPLES II 4 3 Fig.5.11. Isotachop
Page 128 and 129: REFERENCES 113 Example I. As exampl
Page 130 and 131: INSTRUMENTATION
Page 132 and 133: Chapter 6 Detection systems SUMMARY
Page 134 and 135: THERMOMETRIC RECORDING 6.1.3. Combi
Page 138 and 139: THERMOMETRIC RECORDING /2 Fig.6.1.
Page 140 and 141: THERMOMETRIC RECORDING T t l Fig.6.
Page 142 and 143: THERMOMETRIC RECORDING 7 127 It-- I
Page 144 and 145: THERMOMETRIC RECORDING a concentrat
Page 146 and 147: HIGH-FREQUENCY CONDUCTIVITY DETECTI
Page 148 and 149: CONDUCTIVITY DETECTION 133 Fig.6.9.
Page 150 and 151: CONDUCTIVITY DETECTION 135 The d.c.
Page 152 and 153: CONDUCTIVITY DETECTION 137 Hi V 1kR
Page 154 and 155: CONDUCTIVITY DETECTION Fig.6.12. Co
Page 156 and 157: CONDUCTIVITY DETECTION Fig.6.14. Th
Page 158 and 159: CONDUCTIVITY DETECTION 143 6.4.4. T
Page 160 and 161: CONDUCTIVITY DETECTION 145 contact
Page 162 and 163: CONDUCTIVITY DETECTION 147 We can n
Page 164 and 165: 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
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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- 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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