Frans_M_Everaerts_Isotachophoresis_378342.pdf

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DISCUSSION 25 Fig.2.11. Survey of the four main electrophoretic techniques: (a) zone electrophoresis; (b) movingboundary electrophoresis; (c) isotachophoresis; (d) isoelectric focusing. X indicates the position where the sample is usually introduced. For further information, see text. cationic species D. h Fig.2.11 a, the situation in a zone electrophoretic system after a certain time is shown. The background electrolyte AE is present in the whole system, and the anionic species B and C have migrated in the direction of the anode. Anionic species B, which has a higher mobility, has covered a greater migration length. Cationic
26 PRINCIPLES OF ELECTROPHORETIC TECHNIQUES species D has migrated in the direction of the cathode. Fig.2.1 lb shows the movingboundary procedure. The sample mixture is introduced into the cathode compartment and the leading electrolyte AE fills the separation chamber and the anode compartment. The cationic species D thus remains in the cathode compartment and the anionic species B and C, partially separated, migrate behind the leading electrolyte AE. Note that the pure zone B and the mixed zone B+C have the counter ionic species of the leading electrolyte E. The first boundary (with a velocity V,) is sharp (according to the isotachophoretic condition), whereas the separation boundary (with a velocity V*) is not sharp, as in zone electrophoresis. The zone velocities v* and V1 are different. Fig.2.1 lc shows the separation procedure for an isotachophoretic system. The sample is introduced between the leading electrolyte AE and the terminating solution TE. The anionic species B and C are separated and migrate between the terminator T and the leading ion A. All zones have equal velocities, and contain the same counter ionic species E. The cationic species D of the sample has migrated to the cathode. In Fig.2.1 Id, the sample is introduced in an isoelectric focusing system. The point of injection is not important. The ionic species are eventually separated according to their plvalues and are concentrated on spots where the pH values are identical with their pl values, their net charges and velocities then being zero. B and C have migrated towards the anode (lower pH) while the cationic species D has migrated towards the cathode (higher pH). Only amphiprotic substances can be separated by this method.
Page 2 and 3: Journal of Chromatogaphy Library -
Page 4 and 5: Journal of Chromatography Library -
Page 6 and 7: Dedicated to Pr0f.Dr.h. A.I.M. Keul
Page 8 and 9: Contents Preface ..................
Page 10 and 11: CONTENTS IX 6.4.2. The d.c. method
Page 12 and 13: CONTENTS XI 12.2.2. Applications ..
Page 14 and 15: It is very well known that charged
Page 16 and 17: Chapter 1 Historical review SUMMARY
Page 18 and 19: HISTORICAL REVIEW 3 Independently i
Page 20 and 21: THEORY
Page 22 and 23: Chapter 2 Principles of electrophor
Page 24 and 25: MOVINGBOUNDARY ELECTROPHORESIS a S
Page 26 and 27: MOVINGBOUNDARY ELECTROPHORESIS 0 iu
Page 28 and 29: ISOTACHOPHORESIS 13 2.4. PRINCIPLE
Page 30 and 31: ISOTACHOPHORESIS 15 As the anionic
Page 32 and 33: ISOTACHOPHORESIS velocities, so tha
Page 34 and 35: ISOTACHOPHORESIS 19 d.t ’.t I Fig
Page 36 and 37: ISOTACHOPHORESIS 21 t Fig.2.8. Isot
Page 38 and 39: ISOELECTRIC FOCUSING 23 The four is
Page 42 and 43: Chapter 3 Concept of mobility SUMMA
Page 44 and 45: IONIC MOBILITY AND IONIC EQUIVALENT
Page 46 and 47: EFFECTIVE IONIC MOBILITY 31 obtain
Page 48 and 49: EFFECTIVE IONIC MOBILITY 33 compari
Page 50 and 51: EFFECTIVE IONIC MOBILITY Fig.3.4. R
Page 52 and 53: DETERMINATION OF IONIC MOBILITIES 3
Page 54 and 55: DETERMINATION OF IONIC MOBILITIES 3
Page 56 and 57: Chapter 4 Mathematical model for is
Page 58 and 59: GENERAL EQUATIONS 43 anionic specie
Page 60 and 61: GENERAL EQUATIONS 4.2.1. Equilibriu
Page 62 and 63: GENERAL EQUATIONS 41 i Similar equa
Page 64 and 65: GENERAL EQUATIONS 49 Uthzone (U-llt
Page 66 and 67: GENERAL EQUATIONS 4.2.4. Modified O
Page 68 and 69: GENERAL EQUATIONS TABLE 4.2 REDUCED
Page 70 and 71: MATHEMATICAL MODEL FOR THE STEADY S
Page 84 and 85: VALIDITY OF THE ISOTACHOPHORETIC MO
Page 90 and 91:
VALIDITY OF THE ISOTACHOPHORETIC MO
Page 92 and 93:
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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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
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
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DISTURBANCES CAUSED BY H+ AND OH 25
Page 272 and 273:
Page 274 and 275:
DISTURBANCES CAUSED BY H+ AND OH- 2
Page 276 and 277:
Page 278 and 279:
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
Page 288 and 289:
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
Page 302 and 303:
Page 304 and 305:
Page 306 and 307:
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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Page 316 and 317:
SEPARATION USING CONDUCTIVITY AND W
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Page 320 and 321:
Page 322 and 323:
Page 324 and 325:
Page 326 and 327:
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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Page 342 and 343:
Page 344 and 345:
Page 346 and 347:
Page 348 and 349:
Page 350 and 351:
SEPARATION OF SMALL PEPTIDES reprod
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Chapter I4 Separation of nucleotide
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Page 356 and 357:
Page 358 and 359:
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Page 362 and 363:
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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Page 374 and 375:
Page 376 and 377:
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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Page 384 and 385:
Page 386 and 387:
Page 388 and 389:
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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