Frans_M_Everaerts_Isotachophoresis_378342.pdf

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I SEPARATION USING A THERMOCOUPLE AS DETECTOR 29 1 72 __ -1 12 i- 1 h -1 Fig.ll.7. Isotachopherogram of a test mixture of cations carried out in the operational system at pH 5.4 with water and deuterium oxide as solvents (Table 11.3). A conductivity detector (a.c. method) and a W adsorption detector (256 nm) were applied 1 = K+; 2 = BaZ+; 3 = Na+; 4 = (CH,), N+; 5 = Pb2+; 6 = C,H,N+-CH,-CO-NH-NH, ; 7 = Tris'; 8 = histidid; 9 = creatinine*; 10 = bemidine+; 11 = e-aminocaproic acid+; 12 = raminobutyric acid+. The amplification of both the UV absorption detector and the conductivity detector was not changed. A = Increasing UV absorption, R = increasing resistance; t = time. TABLE 11.7 DIFFERENCES IN THE EFFECTIVE MOBILITIES OF SOME CATIONS IN THE SOLVENTS WATER AND DEUTERIUM OXIDE The values (h) refer to the step heights (mm) as measured in the linear trace of the conductivity detector. Ionic species hHZO hDzO Ionic species hH,O hD,O f, K+ 0 0 Tris' 51.0 54.8 Baa+ 8.8 9.1 Histidine' 58.5 65.6 Na + 14.2 16.9 Creatinine' 65.6 78.0 Tma' 20.8 23.8 Benzidine' 81.3 98.1 Pb2+ 25.4 34.8 e-Aminocaproic acid+ 109.2 130.6 C,H,N'-CH,-CO-NH-NH, 39.4 44.8 y-Aminobutyric acid+ 115.6 149.2
292 SEPARATION OF CATIONIC SPECIES IN AQUEOUS SOLUTIONS TABLE 11.8 RELATlVE STEP HEIGHTS OF CATIONS IN THE OPERATIONAL SYSTEM LISTED IN TABLE 11.3 WITH WATER AND DEUTERIUM OXIDE AS THE SOLVENTS For the experiments with deuterium oxide, the same amounts of electrolytes as in the experiments with water were dissolved. The accuracy is better than 4%. The values given are to be used only for the identification of cations in isotachophoretic analyses in the operational systems given. The potassium ion (leading ion) has a relative step height of 0, while the sodium ion has a relative step height of 100. The current was stabilized at 90 pA. - - = No UV absorbance; - = not measured. Ionic species H*O lOOh - uv absorbance hNa D,O lOOh uv - absorbance Amine, butyl Amine, diethanol Amine, ethanol Amine, Zethylaminoethyl Amine, octyl Amine, triethanol Amine, triethyl 2-Amino-2-methyl- 1,3-propanediol Amphetamine Arginine Ammonium, tetrabutyl Ammonium, tetraethyl Ammonium, tetramethyl Barium(I1) Benzidine Benzidine, 3,3-methoxy Butyric acid, 3-amino Cadmium(I1) Calcium(I1) Caproic acid, 2-amino Chromium(II1) Cobalt(I1) 2,4,6-Collidine Copper(1I) Creatinine Diamine, trimethylene Diamine, o-phenylene Girard reagent D Girard reagent P Girard reagent T Guanidine His tidine Hydrazine Hy droxylamine Imidazole Iron(I1) Lanthanum(II1) Lead(I1) 225 29 2 159 51 367 290 290 343 350 422 635 3 04 147 59 576 814 81 3 142 86 772 175 114 315 169 462 35 557 280 277 280 83 411 59 134 119 108 109 179 _ - 165 - 350 - - 313 335 409 590 29 1 142 54 580 - 881 154 81 768 197 1 24 290 237 460 20 550 260 265 252 95 388 64 132 122 128 138 210
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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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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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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
Page 284 and 285: PURIFICATION OF THE TERMINATOR 7 4
Page 286 and 287: REFERENCES 271 AS an example, the c
Page 288 and 289: Chapter 10 Quantitative aspects SUM
Page 290 and 291: THERMOMETRIC MEASUREMENTS 215 vj c
Page 292 and 293: THERMOMETRIC MEASUREMENTS TABLE 10.
Page 294 and 295: CONDUCTIMETRIC MEASUREMENTS factor
Page 296 and 297: CONCLUSION 28 1 Tables 10.3-10.5 wa
Page 298 and 299: Chapter 11 Separation of cationic s
Page 300 and 301: SEPARATION USING A THERMOCOUPLE AS
Page 308 and 309: SEPARATION USING CONDUCTIVITY AND U
Page 310 and 311: Chapter I2 Separation of anionic sp
Page 312 and 313: SEPARATION USING A THERMOMETRIC DET
Page 316 and 317: SEPARATION USING CONDUCTIVITY AND W
Page 326 and 327: Chapter 13 Amino acids, peptides an
Page 328 and 329: AMINO ACIDS TABLE 13.1 OPERATIONAL
Page 330 and 331: AMINO ACIDS 8- ?- 6- 5- 4- 3- 2- f-
Page 332 and 333: AMINO ACIDS 317 TABLE 13.5 STEP HEI
Page 334 and 335: AMINO ACIDS Fig.13.3. Schematic dia
Page 336 and 337: AMINO ACIDS TABLE 13.6 DETERMINATIO
Page 338 and 339: SEPARATION OF PROTEINS IN AMPHOLYTE
Page 350 and 351: SEPARATION OF SMALL PEPTIDES reprod
Page 352 and 353: Chapter I4 Separation of nucleotide
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SEPARATION USING A THERMOMETRIC DET
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SEPARATION USING CONDUCTIVITY AND U
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SEPARATION USING CONDUCTIVITY AND U
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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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