Frans_M_Everaerts_Isotachophoresis_378342.pdf

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CONDUCTIVITY DETECTION Fig.6.14. The d.c.-a.c. converter. 100 kR 10 Ma, although between 10 and 50 kR a fairly linear response can be obtained. With the converter, as shown in Fig.6.14, a maximal potential difference of 10 V can be measured between two points, which has a maximal common mode potential of 6 kV with respect to earth. The impedance between the two points, at the input of the circuit shown in Fig.6.14, is greater than 10l2 !d and the common mode impedance is greater than 10’’ R. The junction-field effect transistor (FET) is used as a source follower. This junction- FET is supplied by a battery, which eventually can be replaced with an electronic circuit, although it is used here in order to minimize both the leak current towards earth (discussed in section 6.6.4) and the parasitic capacitance towards earth. An advantage is that the supply current of the circuit given is very small (
142 DETECTION SYSTEMS where Yis the voltage on the potential recorder, and Int and Zero are the positions of the controls ‘Int’ and ‘Zero’, respectively. The pA meter indicates 50 pA if V, = 0 V and 100 pA if Vh = 10 V. The accuracy in measuring Vin is better than 50 mV. If the ambient temperature changes from 10 to 35 C, the difference in the measured value of Yb, which of course does not change during this procedure, is less than 20 mV. In Fig.6.15, an isotachopherogram of a test mixture of anions in the operational system at pH 6 (Table 12.1) is given. In the conclusion of this chapter, more attention is paid to this method of detection. It can be seen that Fig.6.15 is comparable with the linearized isotachopherogram obtained with the a.c. conductimetric circuit, shown in Fig.6.18; in fact, we did not find any characteristic differences. 10 10 d.C. 6 - rO I 7T/J1 10 Fig.6.15. Isotachopherogram of a test mixture of anions in the operational system at pH 6 (Table 12.1), as derived from the d.c.-a.c. converter in combination with the circuit shown in Fig.6.18. For comparison, an isotachopherogram obtained with the circuit shown in Fig.6.18 is also given. This test mixture was applied in almost all isotachopherograms, for pattern recognition, in order to show the various effects that may occur if insufficient precautions are taken. The leading electrolyte consists of 0.01 N hydrochloric acid (pro analysi grade) and histidine, adjusted to pH 6; 0.05% (w/w) of Mowiol (polyvinylalcohol) was added to the electrolyte. Glutamic acid (0.005 N) was used as the terminating electrolyte, adjusted to pH cn. 6 by addition of Tris. 1 = Chloride; 2 = sulphate; 3 = chlorate; 4 = chromate; 5 = malonate; 6 = pyrazole-3,5-dicarboxylate; 7 = adipate; 8 = acetate; 9 = P-chloropropionate; 10 = glutamate. R = Increasing electric resistance; V = increasing potential gradient; t = time. i 8 - a.c. 1
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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
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 136 and 137: THERMOMETRIC RECORDING Potential gr
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 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
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
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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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