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Chromatography:

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Bibliographic Details
Format:	Book
Language:	English
Published:	New York Reinhold 1967
Edition:	2. ed.
Series:	Reinhold chemistry textbook series
Subjects:	Chromatographie
Online Access:	Inhaltsverzeichnis
Physical Description:	XLI, 851 S. Ill., graph. Darst.

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245	1	0	\|a Chromatography \|c Erich Heftmann [Ed.]
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adam_text	Contents Collaborators................................................................... jx Foreword to the Second Edition by Edgar Lederer.......................... xiii Foreword to the First Edition by Arne Tiselius................................ xv Preface to the Second Edition by Erich Hetimann........................... xvii Preface to the First Edition by Erich Heftmann............................... xix PART ONE Fundamentals of Chromatography CHAPTER l Historical Introduction L. Zechmeisfer California Institute of Technology Pasadena, California References....................................................................... 9 CHAPTER 2 Differential Migration Methods of Analysis................. 11 Harold H. Strain Argonne National Laboratory Argonne, Illinois 1. Evolution and Concepts........................................................... II The Nature of Chromatography and Related Methods............................. II Basic Operations............................................................... II Basic Nomenclature............................................................ I - Modifications of Chromatography.............................................. I ¦? Significance of Chromatography................................................ I -5 Theories of Chromatography ................................................... 14 xxii CONTENTS Concept of Differential Migration............................................... 14 Definitions..................................................................... 5 Chromatography............................................................... 5 Zone Electromigration.......................................................... 5 Definition of Zone Electromigration............................................. 16 Continuous Zone Electromigration.............................................. 16 Definition of Continuous Zone Electromigration................................. 17 2. Ramifications of Chromatography and Zone Electromigration...................... 17 Operational Considerations....................................................... 17 Principal Uses.................................................................. 17 Detection Methods............................................................. 18 R Values....................................................................... 18 Mixed Chromatograms and Chromatographic Sequences......................... 19 Condition of Substances to be Examined........................................ 19 Solvents, Sorbents, and Stabilizers .............................................. 19 Chromatographic Systems...................................................... 20 Specific Applications of Differential Migration Methods ......................... 20 References....................................................................... 21 CHAPTER 3 Theory of Chromatography.................................... 23 J. Calvin Giddings University of Utah Salt Lake City, Utah 1. Introduction...................................................................... 23 2. Mechanism of Zone Migration.................................................... 24 3. Zone Spreading and the Concept of Nonequilibrium ............................... 27 4. Chromatography as a Random Walk.............................................. 31 5. Separation ....................................................................... 38 References...................................... ................ 41 CHAPTER 4 Adsorption....................................................... 43 L R. Snyder Union Oil Company of California Brea, California 1. Introduction...................................................................... 43 2. General Aspects of Adsorption.................................................... 43 3. The Adsorbent................................................................... 46 Silica........................................................................... 49 Alumina....................................................................... 52 Charcoal....................................................................... 53 Other Adsorbents............................................................... 54 Modified Adsorbents........................................................... 56 4. The Solvent...................................................................... 57 5. The Sample: Relative Separation Order Versus Molecular Structure................. 62 Group Adsorption Energies....................................................... 63 CONTENTS xxjjj Sample Localization.............................................................. 66 Intramolecular Electronic Effects.................................................. 68 Intramolecular Steric Effects...................................................... 69 Sample Planarity............................................................... 69 Steric Hindrance of Adsorption................................................. 70 Chemical Interaction Between Adjacent Sample Groups.......................... 71 Concerted Adsorption of Adjacent Groups on a Single Adsorption Site........... 71 Restricted Internal Rotation.................................................... 72 Sample Conjugation or ir-Electron Quantum Effects................................ 72 6. Temperature Effects.............................................................. 73 7. A General Model for Correlating Sample Separation Order with Structure.......... 74 References ....................................................................... 76 CHAPTER 5 Techniques of Liquid Column Chromatography............ 80 L. R. Snyder Union Oil Company of California Brea, California 1. Introduction...................................................................... 80 2. Fundamental Modes of Column Chromatography................................. 81 Column Development ............................................................ 82 Frontal Analysis.................................................................. 82 Displacement..................................................................... 82 Elution........................................................................... 83 Coupled Columns................................................................ 87 Combination of Techniques....................................................... 87 3. Some Factors That Affect Column Efficiency....................................... 87 Column Dimensions.............................................................. 87 Particle Size of the Column Packing............................................... 88 Pore Diameter of the Column Packing............................................. 90 Adsorbent Activity............................................................... 90 Packing the Column.............................................................. 90 Selection of Solvents.............................................................. 91 Solvent Flow Rate................................................................ 92 Column Temperature............................................................. 92 Sample Application............................................................... 92 Some Alternative Forms of Column Chromatography.............................. 93 4. Sample Detection................................................................. 93 Detection on the Column......................................................... 94 Detection in the Fractions......................................................... 94 Continuous Detection in the Eluate................................................ 94 5. Apparatus........................................................................ 96 Gradient Devices................................................................. 97 Solvent Flow Control............................................................. !05 Sample Injection Devices.......................................................... 05 Columns......................................................................... l06 Fraction Collectors............................................................... 107 6. Miscellaneous Technical Aspects.................................................. 08 Sample Recovery................................................................. 08 Solvent Purification............................................................... 08 CONTENTS Column Saturation and Displacement Effects...................................... OS Reusable Columns................................................................ l09 7. Future Trends in Liquid Column Chromatography . ............................ 109 References ....................................................................... 10 CHAPTER 6 Theoretical Basis of Partition Chromatography?............ 113 Roy A.Keller University of Arizona Tucson, Arizona J. Calvin Giddings University of Utah Salt Lake City, Utah 1. Introduction...................................................................... 3 2. The Liquid-Liquid Partition Model of Chromatography............................ 113 3. Extension and Criticism of the Partition Model .................................... 115 4. Plate Height in Gas Chromatography.............................................. IIS 5. Migration Parameters: R Values................................................... 121 6. Role of the Support and the Stationary Liquid Phase............................... 123 7. Mechanism of Paper Chromatography............................................. 125 8. The Significance of RF,RM, and R ................................................ 130 References ....................................................................... 134 CHAPTER 7 Techniques of Paper Chromatography....................... 139 Karel Macek Research Institute for Pharmacy and Biochemistry Prague, Czechoslovakia Translated by E. Heftmann 1. Introduction .................................................................... 139 2. Preparation of Sample........................................................... 140 Solution...................................................................... 140 Extraction.................................................................... 140 Removal of Ballast Material................................................... 140 Preparation of Derivatives..................................................... 140 3. Paper........................................................................... 141 Papers of Pure Cellulose....................................................... 141 Modified Cellulose Papers..................................................... 141 Papers of Other Materials ..................................................... 142 4. Sample Application.............................................................. 143 5. Solvent Systems................................................................. 143 Aqueous Stationary Phase..................................................... 144 Stationary Hydrophilic Organic Solvent........................................ 145 Reversed Phases (Stationary Hydrophobic Solvent)............................. 145 6. Tank and Development.......................................................... 146 Techniques with Various Flow Directions......................................... 146 This work was supported by National Science Foundation GP-4643 and N1H grant (255-512592). CONTENTS xxv Descending Development...................................................... ]46 Ascending Development....................................................... 146 Horizontal Development...................................................... 147 Radial Development .......................................................... 148 Multiple Chromatography....................................................... 149 Multiple Development......................................................... 149 Two-Dimensional Chromatography............................................ 149 Multidimensional Chromatography............................................ 149 Special Procedures............................................................... 149 Overrun Development......................................................... 149 Slotted Paper................................................................ 149 Other Techniques............................................................. 149 7. Detection....................................................................... 150 Physical Detection Methods................................................... 150 Chemical Detection Methods.................................................. 151 Enzymatic and Biological Detection Methods................................... 151 8. Identification of Substances...................................................... 151 Systematic Analysis........................................................... 152 9. Quantitative Analysis............................................................ 154 In Situ Methods................................................................. 154 Visual Estimation............................................................. 154 Photometric Methods......................................................... 155 Other Methods................................................................ 155 Methods Following Elution...................................................... 155 10. Preparative Chromatography.................................................... 156 Arrangements with Free Paper Sheets.......................................... 156 Chromatopile................................................................. 157 Chromatopack................................................................ 157 Rolls of Filter Paper........................................................... 157 Continuous Chromatography.................................................. 157 11. Radioisotope Methods........................................................... 158 Autoradiography.............................................................. 158 Radiometric Methods ......................................................... 158 Use of Radioisotope Techniques............................................... 160 References...................................................................... 160 CHAPTER 8 Techniques of Thin-Layer Chromatography................. 165 Egon Stahl Universitat des Saarlandes Saarbrijcken, Saar, Germany Helmut K. Mangold University of Minnesota Austin, Minnesota 1. Introduction...................................................................... 65 2. Basic Equipment ................................................................. 65 3. Coating Materials................................................................ 69 4. Preparation of Plates.............................................................. 70 5. Solvents and Development............................-............................ ?4 6. Visualization and Documentation................................................. 176 xxvi CONTENTS 7. Quantitative Analysis............................................................. ° 8. Preparative and Combined Techniques ............................................ 179 References....................................................................... l8° CHAPTER 9 Techniques of Gas Chromatography......................... 182 C. A. M. G. Cramers Technological University Eindhoven, The Netherlands A. . M. Keulemans Technological University Eindhoven, The Netherlands Harold M. McNair /nfernafi ona/ Varian Aerograph Walnut Creek, California 1. Introduction...................................................................... 82 Stationary and Mobile Phases................................................... 82 Manner of Execution........................................................... 182 2. Theory........................................................................... 84 Location of a Peak in a Chromatogram............................................ 184 Partition Coefficient, Capacity Ratio, Retardation Factor........................ 184 Peak Broadening ................................................................. 186 Chromatographic Separations, Resolution......................................... 187 3. Optimali/ation................................................................... 190 Separation ....................................................................... 190 The Separation Factor.......................................................... 190 The Capacity Ratio............................................................. 193 The Plate Height............................................................... 195 Analysis Time.................................................................... 195 Sample Size ...................................................................... 195 4. Instrumental Aspects............................................................. 197 Support.......................................................................... 197 Carrier Gas ...................................................................... 198 Viscosity....................................................................... 199 DitTusivity ..................................................................... 199 Thermal Conductivity.......................................................... 199 Safety.......................................................................... 199 Flow Control .................................................................. 199 Temperature Control............................................................. 200 Sampling Devices................................................................. 200 Detectors......................................................................... 201 Thermal Conductivity Detector................................................. 202 Flame lonizmion Detector...................................................... 204 5. Qualitative Analysis.............................................................. 205 Retention Volumes............................................................... 205 Special Plots of Retention Volumes................................................ 206 6. Quantitative Analysis............................................................. 207 Automatic Integrators............................................................ 208 References....................................................................... 209 CONTENTS xxvii CHAPTER 10 Theory of Electrophoresis..................................... 210 R. J. Wieme University of Ghent Ghent, Belgium 1. Introduction...................................................................... 210 2. Physical Basis of Electrophoresis.................................................. 211 The Electric Double Layer of Small Ions........................................... 211 The Electric Double Layer of Large Ions........................................... 212 Equations for Electrophoretic Movement of Small Ions............................. 213 Equations for Electrophoretic Movement of Large Ions ............................ 215 Equation of Electroosmotic Flow.................................................. 216 3. Basic Forms of tlectrophoresis.................................................... 218 Moving Boundary Free Electrophoresis............................................ 218 Boundaries in Strong Electrolytes............................................... 219 Conditions a! the Moving Boundary.......................................... 220 Conditions at the Stationary Boundary........................................ 221 Application to Frontal Electrophoresis.......................................... 221 Conclusion..................................................................... 223 Zone Electrophoresis in a Supporting Medium..................................... 223 Survey of Supporting Media.................................................... 223 Distortions of Zone Shape...................................................... 224 Uneven Temperature Distribution............................................ 224 Uneven Hydraulic Flow...................................................... 225 Faults in the Support......................................................... 225 Changes in Electric Conductivity or pH at the Zone............................ 225 Electroosmosis............................................................... 225 Other Causes ................................................................ 226 Electrochromatography......................................................... 226 4. Separability in Electrophoresis.................................................... 228 Linear Thermal Diffusion......................................................... 228 Microheterogeneity............................................................... 229 Electrodiffusion .................................................................. 231 Eddy Migration .................................................................. 232 Electrosorptive Spreading......................................................... 232 Optimizing Electrophoretic Conditions............................................ 233 5. Special Aspects................................................................... 235 Temperature Effects in High-Voltage Electrophoresis............................... 235 Measurement of Electrophoretic Mobility ......................................... 237 Conditions for Valid Measurement.............................................. 239 Test for Field Homogeneity..................................................... 240 Test for Field Constancy........................................................ 240 Measurement of Absolute Mobility ............................................. 240 Measurement of Relative Mobility.............................................. 240 Conversion of ma into Free Mobility.......................................... 241 Tortuosity................................................................... 242 Constriction................................................................. 242 Ion Retardation.............................................................. 243 Zone Sharpening................................................................. 244 Discontinuous Buffer Systems. Disc Electrophoresis............................... 245 Molecular Dimensions from Electrophoresis....................................... 247 References ....................................................................... 249 xxviii CONTENTS CHAPTER 11 Techniques of Electrophoresis................................ 252 Heribert Michl Hochschule fur Boden/cu/rur Vienna, Austria Translated by £. Heffmann 1. Analytical Methods............................................................... 252 Electrophoresis on Paper Strips and Similar Carriers............................... 252 High-Voltage Electrophoresis................................................... 253 Apparatus with Liquid Heat Exchangers...................................... 253 Apparatus with Solid Heat Exchangers........................................ 255 Combination of High-Voltage Electrophoresis and Paper Chromalography..... 258 Electrolytes for High-Voltage Electrophoresis................................. 258 Selected Applications of High-Voltage Electrophoresis......................... 260 Low-Voltage Electrophoresis ................................................... 269 Electrophoresis in Gels............................................................ 271 Immunoelectrophoresis........................................................... 275 2. Preparative Methods.............................................................. 277 Discontinuous Procedures ........................................................ 277 Electrophoresis in Slabs of Porous Material ................................... 277 Electrophoresis in Columns................................................... 278 Continuous Electrophoresis....................................................... 280 References....................................................................... 282 CHAPTER 12 Principles of Ion Exchange.................................... 287 Harold F. Walton University of Colorado Boulder, Colorado 1. Introduction...................................................................... 287 2. inorganic Ion Exchangers......................................................... 287 Aluminosilicates.................................................................. 287 Hydrous Oxides.................................................................. 288 Other Inorganic Exchangers....................................................... 289 3. Ion-Exchange Resins ............................................................. 289 Electron-Exchange Resins......................................................... 291 Commercial Resins............................................................... 292 Particle Size.................................................................... 292 Cross-Linking.................................................................. 294 Porous Resins.................................................................. 294 4. Cellulose Ion Exchangers ......................................................... 294 5. Physical Properties of Ion-Exchange Resins........................................ 295 Water Uptake, Swelling, Electrolyte Uptake....................................... 295 Ion-Exchange Equilibrium: Selectivity............................................. 297 Theories of Selectivity............................................................. 299 Thermodynamic Interpretation of Ion-Exchange Selectivity......................... 302 Ion-Exchange Kinetics............................................................ 305 CONTENTS xxix 6. Ion-Exchange Columns........................................................... 30d Basic Theory..................................................................... 30f, Displacement .................................................................. 107 Elution: Theoretical Plate Concept.............................................. 3(17 Efficiency of Elution Separation................................................. 313 Practical Operation............................................................... 317 7. Complex Ions in Ion Exchange.................................................... 31S General Considerations........................................................... 31S Anion Exchange of Metal Complexes.............................................. 31S Ligand Exchange................................................................. 319 8. Ion Exclusion and Ion Retardation................................................ 321 References....................................................................... 322 CHAPTER 13 Techniques and Applications of Ion-Exchange Chromatography ........................................... 325 Harold F. Walton University of Colorado Boulder, Colorado 1. The Column...................................................................... 325 Principles and Analytical Applications of Ion Exchange............................ 326 2. Collection and Analysis of Effluents............................................... 327 Batch Operation.................................................................. 327 Continuous Monitoring........................................................... 328 3. Typical Analytical Applications: Inorganic......................................... 329 Determination of Total Electrolyte Concentration.................................. 329 Separation of Interfering Ions..................................................... 330 Concentration of Traces .......................................................... 331 Separation of Anionic Metal Complexes........................................... 332 Chromatographic Separations of Cations and Anions.............................. 333 Cations........................................................................ ^i3 Anions......................................................................... 335 4. Typical Analytical Applications: Organic.......................................... 336 Separation of Carboxvlic Acids.................................................... 336 Separation of Organic Bases....................................................... 337 Nonionic Separations: Solubilization and Salting-Out Chromatography........ 337 5. Paper and Thin-Layer Chromatography........................................... 338 Separations of Inorganic Ions..................................................... 339 Biochemical Separations.......................................................... 340 References....................................................................... 340 CHAPTER 14 Gel Filtration................................................... 343 Bertil Gelotte Pharmacia Fine Chemicals Uppsala, Sweden Jerker Porath Institute of Biochemistry Uppsala, Sweden 1. Introduction...................................................................... 343 2. Types of Gels..................................................................... 344 xxx CONTENTS 3. Techniques....................................................................... 347 Column Techniques.............................................................. 347 Thin-Layer Techniques........................................................... 350 4. Various Phenomena Encountered in Gel Filtration................................. 351 Molecular Exclusion.............................................................. 351 Exclusion Behavior Modified by Solute-Solute Interaction.......................... 354 Aromatic Sorption................................................................ 357 Other Kinds of Sorption and Exclusion Phenomena................................ 360 Utilization of Various Interactions to Improve Fractionation....................... 363 5. Theoretical Aspects of Molecular Sieving in Gels................................... 364 References....................................................................... 368 PART TWO Applications of Chromatography CHAPTER 15 Chromatography of Amino Acids and Peptides........... 373 G. Pataki Robapharm A. G. Basel, Switzerland £. Baumann Varian Aerograph A.G. Basel, Switzerland U. P. Ge/ger University of Basel Basel, Switzerland P. Jenkins Varian Aerograph A.G. Basel, Switzerland U. Kupper Varian Aerograph A.G. Basel, Switzerland Translated by E. Hefimann 1. Introduction...................................................................... 373 2. Paper and Thin-Layer Chromatography........................................... 373 Chromatography of Free Amino Acids............................................ 373 Chromatography of Dinitrophenyl Amino Acids................................... 378 3. Electrophoresis................................................................... 385 Separation of Amino Acids ....................................................... 385 Separation of Peptides............................................................ 387 4. Ion-Exchange Chromatography and Other Methods of Column Chromatography ... 389 5. Gas Chromatography............................................................. 395 Separation of Amino Acids ....................................................... 395 Separation of Peptides............................................................ 398 References....................................................................... 400 CONTENTS xxxj CHAPTER 16 Chromatography of Proteins................................. 405 Hugo Fasold Physiologisch-Chemisches Instifut der Universit af Wurzburg Wurzburg, Germany Gerd Gundlach Laboraforium der Urologischen Universif atsklinik Homburg, Soar, Germany Friedrich Turba, deceased Formerly, Physiologisch-Chemisches Institut der Universit at Wurzburg Wurzburg, Germany Translated by E. Heftmann 1. introduction...................................................................... 405 2. Ion-Exchange Chromatography...................................................407 Principles........................................................................407 Cellulose Exchangers............................................................. 410 Carboxymethyl (CM) Cellulose................................................. 410 Enzymes..................................................................... 410 Hemoglobins................................................................ 411 Serum Proteins............................................................... 412 Hormones................................................................... 412 Proteins from Secretions, Cells, and Organs................................... 412 Cellulose Phosphate (CP)....................................................... 414 Diethylaminoethyl (DEAE) Cellulose ........................................... 414 Enzymes..................................................................... 414 Hemoglobins................................................................ 419 Serum Proteins............................................................... 419 Hormones................................................................... 421 Proteins from Cells and Organs............................................... 422 Virus Proteins................................................................ 424 Aminoethyl (AE) Cellulose..................................................... 424 Triethylaminoethyl (TEAE) Cellulose...........................................424 ECTEOLA Cellulose...........................................................425 Sulfomethyl (SM) Cellulose..................................................... 426 Dextran Exchangers.............................................................. 426 DEAE Sephadex............................................................... 426 Cation Exchangers............................................................. 428 Resin Exchangers................................................................. 429 Carboxyl Resin (Amberlite RC-50).............................................. 429 Sulfonic Acid Resins (Dowex 50, Zeo-Karb 225).................................433 Amine Resin (Dowex 2)........................................................ 434 3. Adsorption Chromatography ..................................................... 434 Principles........................................................................ 434 Applications...................................................................... 436 4. Partition Chromatography........................................................ 438 Principles........................................................................ 438 Applications...................................................................... 438 xxxii CONTENTS 5. Gel Filtration .................................................................... 439 Principles ........................................................................ 439 Dextran Gels ................................................................ 439 Other Gels................................................................... 441 Applications...................................................................... 441 6. Column Chromatography and Gel Filtration in the Determination of Protein Structure................................................................. 444 7. Electrophoresis in Columns....................................................... 445 Solid Supports.................................................................... 445 Principles...................................................................... 445 Apparatus................................................................... 446 Column Material............................................................. 448 Packing of the Column....................................................... 448 Determination of Column Characteristics..................................... 448 Sample Application.......................................................... 449 Electrophoresis.............................................................. 449 Zone Sharpening............................................................. 449 Electroosmosis............................................................... 450 Elution...................................................................... 450 Applications.................................................................... 450 Density Gradient Electrophoresis.................................................. 453 Apparatus................................................................... 453 Operation................................................................... 453 Examples.................................................................... 454 8. Discussion ....................................................................... 455 References ....................................................................... 458 CHAPTER 17 Chromatography of Lipids.................................... 466 Lindsay J. Morris Unilever Research Laboratory Sharnbrook, Bedford, England Bryan W. Nichols Unilever Research Laboratory Sharnbrook, Bedford, England 1. Introduction...................................................................... 466 2. Preliminary Separations........................................................... 467 3. Separation of Neutral Lipid Classes............................................... 468 Column Chromatography......................................................... 468 Paper and Thin-Layer Chromatography........................................... 469 4. Separation of Phospholipids and Glycolipids....................................... 470 Column Chromatography......................................................... 470 Paper Chromatography........................................................... 472 Thin-Layer Chromatography...................................................... 472 5. Separations of Polar Hydrolysis Products from Lipids.............................. 474 6. Separations of Lipid Hydrolyzates................................................. 475 Liquid-Solid Chromatography .................................................... 476 Liquid-Liquid Chromatography................................................... 480 CONTENTS xxxiii Argentation Chromatography.................................... 4x5 Gas-Liquid Chromatography ........................................... 4Xj( 7. Separations Within Intact Lipid Classes......................................... 493 Liquid-Solid Chromatography......................................... 49} Liquid-Liquid Chromatography...................................... 494 Argentation Chromatography................................................. 495 Gas-Liquid Chromatography ........................................... 49(, 8. Some Practical Considerations in Lipid Chromatography.......................... 497 Stability of Lipids during Chromatography........................................ 497 Detection of Lipids during Chromatography....................................... 498 Monitoring of Column Effluents.............................................. 498 Detection of Lipids on Paper and Thin-Layer Chromatograms................. 499 Quantitation of Lipids Separated by Chromatographic Methods.................... 499 Radiochemical Detection of Lipids................................................ 500 References....................................................................... 501 CHAPTER 18 Chromatography of Terpenes................................ 510 Franiisek Sorm Czechoslovak Academy of Science Prague, Czechoslovakia Vlastimil Herouf Czechoslovak Academy of Science Prague, Czechoslovakia Translated by L. Labler 1. Introduction...................................................................... 510 Definitions and Scope............................................................. 510 History........................................................................... 511 Bibliography..................................................................... 512 2. Precursors of Terpenoids.......................................................... 512 3. Monoterpenes.................................................................... 513 Adsorption Chromatography on Columns......................................... 514 Paper Chromatography of Monoterpenes (Tropolones)............................. 514 Gas-Liquid Chromatography ..................................................... 515 Thin-Layer Chromatography...................................................... 519 4. Sesquiterpenes.................................................................... 521 Adsorption Chromatography on Columns......................................... 521 Partition Chromatography of Sesquiterpenes and Azulenes......................... 522 Ion-Exchange Chromatography of Azulenes....................................... 523 Gas-Liquid Chromatography of Sesquiterpenes.................................... 524 Thin-Layer Chromatography of Sesquiterpenes.................................... 524 5. Diterpenes and Triterpenes........................................................ 527 Column and Thin-Layer Adsorption Chromatography............................. 527 Partition Chromatography........................................................ 528 Gas-Liquid Chromatography ..................................................... 528 6. Carotenoids...................................................................... 529 Adsorption Chromatography on Columns......................................... 5-9 Paper Chromatography........................................................... 531 Thin-Layer Chromatography...................................................... 531 References....................................................................... • - ¦ xxxiv CONTENTS CHAPTER 19 Chromatography of Steroids................................. 539 Erich Heftmann Western Regional Research Laboratory Albany, California, and California Institute of Technology Pasadena, California Herbert H. Wotiz Boston University, School of Medicine Boston, Massachusetts 1. Introduction...................................................................... 539 2. Liquid Column Chromatography.................................................. 539 3. Thin-Layer Chromatography...................................................... 545 General.......................................................................... 545 Sterols and their Esters............................................................ 548 Steroid Sapogenins and Saponins.................................................. 550 Steroid Alkaloids and Glycoalkaloids.............................................. 551 Cardiac Clycosides and Aglycones................................................. 553 Bile Acids and Alcohols........................................................... 554 Corticosteroids and Other C21 Steroids............................................ 555 Androgens and Other C19 Steroids................................................. 559 Estrogens and Other C\|g Steroids.................................................. 559 4. Gas Chromatography............................................................. 561 General.......................................................................... 561 Instrumentation................................................................ 561 Columns....................................................................... 561 Retention Time and Structure................................................... 562 Quantitative Analysis in Biological Fluids....................................... 563 Estrogens ........................................................................ 563 Progesterone and Metabolites..................................................... 565 Urinary Pregnanediol........................................................... 565 Progesterone in Plasma......................................................... 566 17- Ketosteroids................................................................... 566 Testosterone...................................................................... 567 Corticosteroids................................................................... 568 References ..................................................................... 569 CHAPTER 20 Chromatography of Carbohydrates and Related Compounds............................................... 573 Francois Percheron Faculte de Pharmacie Paris, France Translated by E. Heftmann 1. Introduction..................................................................... 573 2. Paper Chromotography........................................................... 573 A laboratory of the Western Utilization Research and Development Division, Agri- cultural Research Service, U.S. Department of Agriculture. Work conducted under a cooperative agreement with the California Institute of Technology. CONTENTS xxxv Techniques..................................................................... 574 Solvent Systems.................................................................. 576 Detecting Reagents .............................................................. 577 Oxidizing Agents................................................................579 Reagents for Glycols........................................................... 5X0 Furfural Reactions............................................................. 581 Amine Reagents ............................................................. 581 Phenolic Reagents............................................................ 582 Mixed Reagents.............................................................. 582 Other Reagents.............................................................. 582 Main Classes of Carbohydrates.................................................... 583 Aldoses and Ketoses......................................................... 583 Oligosaccharides........................................................... 583 Methylated Sugars........................................................... 584 Glycosides................................................................... 585 Sugar Esters................................................................. 585 Acids and Lactones.......................................................... 5X6 Amino Sugars................................................................ 586 Deoxysugars and Related Compounds........................................ 587 Polyols and Cyclitols......................................................... 587 Quantitative Methods............................................................. 587 Determination After Staining the Chromatograms............................... 588 Direct Photodensitometry.................................................... 58X Elution Methods............................................................. 5X8 Determination on Eluates Without Previous Staining ............................ 588 Selected Applications of Paper Chromatography................................... 5XX Isolation, Identification, and Separation of Mono- and Oligosaccharides........ 588 Structural Studies on Oligo- and Polysaccharides.............................. 589 Enzymatic Reactions......................................................... 589 3. Paper Electrophoresis............................................................. 589 4. Column Chromatography......................................................... 593 Adsorption Chromatography ..................................................... 594 Charcoal..................................................................... 594 Clays and Silicates........................................................... 594 Alumina..................................................................... 594 Partition Chromatography........................................................ 595 Ion-Exchange Chromatography................................................... 595 Involving Ion Exchange...................................................... 595 Not Involving Ion Exchange.................................................. 596 Gel Filtration .................................................................... 597 Gas-Liquid Chromatography ..................................................... 597 5. Thin-Layer Chromatography...................................................... 598 Supports..................................................................... 599 Solvent Mixtures............................................................. 599 Applications................................................................. 600 References ....................................................................... 601 xxxvj CONTENTS CHAPTER 21 Chromatography of Alkaloids............................... 606 Karel Art ace k Research Institute for Pharmacy and Biochemistry Prague, Czechoslovakia Translated by E. Heftmann 1. Introduction...................................................................... 606 2. Paper Chromatography........................................................... 606 Detection ........................................................................ 606 Solvent Systems.................................................................. 607 Quantitative Analysis............................................................. 608 3. Thin-Layer Chromatography...................................................... 609 Detection ........................................................................ 609 Sorbents......................................................................... 609 Solvent Systems.................................................................. 609 Quantitative Analysis............................................................. 610 4. Gas Chromatography............................................................. 610 5. Applications...................................................................... 611 Structure Determination.......................................................... 611 Systematic Analysis............................................................... 611 Individual Groups of Alkaloids ................................................... 613 Tobacco Alkaloids............................................................. 613 Paper Chromatography ...................................................... 613 Thin-Layer Chromatography................................................. 613 Gas Chromatography........................................................ 613 Tropine Alkaloids.............................................................. 614 Paper Chromatography ...................................................... 614 Thin-Layer Chromatography................................................. 616 Gas Chromatography........................................................ 616 Opium Alkaloids............................................................... 616 Paper Chromatography ...................................................... 616 Thin-Layer Chromatography................................................. 617 Gas Chromatography........................................................ 618 Ergot Alkaloids................................................................ 619 Paper Chromatography ...................................................... 619 Thin-Layer Chromatography................................................. 620 Rauwoltia Alkaloids............................................................ 621 Paper Chromatography ...................................................... 621 Thin-Layer Chromatography................................................. 622 Other Groups of Alkaloids...................................................... 623 Purine Alkaloids............................................................. 623 Lupine Alkaloids............................................................. 623 Cannabis Alkaloids.......................................................... 623 Veratrum Alkaloids.......................................................... 623 Vinca Alkaloids.............................................................. 623 Harmane Alkaloids.......................................................... 623 Ipecacuanha Alkaloids....................................................... 623 Curare Alkaloids............................................................. 623 Cincona Alkaloids........................................................... 623 Aporphine Alkaloids......................................................... 623 References ....................................................................... 623 CONTENTS xxxvii CHAPTER 22 Column Chromatography of Nucleic Acids and Related Substances......................................... 627 Waldo E. Cohn Oak Ridge National Laboratory Oak Ridge, Tennessee 1. Introduction...................................................................... 627 2. Bases............................................................................. 629 3. Nucleosides...................................................................... 632 4. Nucleotides ...................................................................... 636 Simple Nucleotides............................................................... 636 Sugar Phosphates................................................................. 640 Nucleoside Polyphosphates....................................................... 642 Oligonucleotides.................................................................. 644 5. N ucleic Acids .................................................................... 649 6. Viruses........................................................................... 653 7. Desalting......................................................................... 656 References....................................................................... 657 CHAPTER 23 Chromatography of Chlorophylls and Various Porphyrins........................................... 661 Harold H. Strain Argonne National Laboratory Argonne, Illinois 1. Diversity of Porphyrin Compounds................................................ 661 Variations of Porphyrins.......................................................... 661 2. Chlorophylls..................................................................... 662 Significance of Chlorophylls....................................................... 662 Chlorophylls and the History of Chromatography.................................. 662 Lability of the Chlorophylls....................................................... 662 Extraction of the Chlorophylls.................................................... 663 Chromatographic Procedures for Separation of Chlorophylls and Carotenoids ...... 663 Sorptive Systems ................................................................. 663 Column Chromatography......................................................... 664 Isolation of Separated Chlorophylls............................................... 665 Chlorophylls of Various Plants.................................................... 665 Estimation of the Chlorophylls.................................................... 665 Chromatographic Sequences...................................................... 666 Various Adsorbents............................................................... 666 Paper Chromatography........................................................... 667 Anomalous Separations in Paper.................................................. 669 Thin-Layer Chromatography...................................................... 670 Gas Chromatography............................................................. 670 3. Porphyrins and Hemins........................................................... 670 Properties........................................................................ 670 Column Chromatography......................................................... 671 Paper Chromatography........................................................... 672 This work was sponsored by the U.S. Atomic Energy Commission under contract with the Union Carbide Corporation. CONTENTS XXXVIII Gas Chromatography............................................................. 4. Oxidation Products of Porphyrins................................................. Paper Chromatography........................................................... 5. Bile and Urinary Pigments........................................................ . Column Chromatography......................................................... 6. Complexes of Porphyrins and Proteins............................................. 67 Various Separations.............................................................. 7. Phycobilins....................................................................... 6 Column Chromatography......................................................... r. r 674 References...................................................................... CHAPTER 24 Chromatography of Phenolic Compounds.................. 677 Jeffrey B. Harborne University of Liverpool Liverpool, England 1. Introduction...................................................................... 677 2. Paper Chromatography........................................................... 678 Solvent Systems.................................................................. 678 Color Reactions.................................................................. 680 RF Values........................................................................ 680 Flavonoids....................................................................... 680 Anthocyanins.................................................................... 683 Flavonols and Flavones........................................................... 684 Flavonol Methyl Ethers......................................................... 684 Flavonol Glycosides............................................................ 685 Flavone Glycosides............................................................. 685 Sources of Error.................................................................. 685 3. Paper Electrophoresis............................................................. 686 4. Thin-Layer Chromatography...................................................... 687 General Considerations........................................................... 687 Sorbents and Solvents............................................................. 688 Simple Phenolics................................................................. 690 Flavonoids....................................................................... 691 Hydroxyquinones................................................................. 692 5. Column Chromatography: Large-Scale Separations................................ 693 Comparison of Methods.......................................................... 693 Thick Paper...................................................................... 693 Silica Gel......................................................................... 694 Polyamide........................................................................ 694 6. Gas-Liquid Chromatography ..................................................... 695 References....................................................................... 696 CHAPTER 25 Chromatography of Vitamins................................ 699 Goicfiiro Katsui Eisai Co., Ltd. Tokyo, Japan 1. Introduction...................................................... 599 2. Thin-Layer Chromatography.......................................... 699 Detection Methods............................................. 699 CONTENTS xxxix Fat-Soluble Vitamins............................................................. 700 Vitamin A ..................................................................... 700 Vitamin D..................................................................... 703 Vitamin E...................................................................... 704 Vitamin K and Coenzyme Q.................................................... 705 Water-Soluble Vitamins........................................................... 706 Vitamin B,..................................................................... 706 Vitamin B2..................................................................... 706 Vitamin B6..................................................................... 706 Niacin and Niacinamide........................................................ 708 Pantothenic Acid and Pantothenol.............................................. 708 Vitamin B l2.................................................................... 708 Other Vitamins................................................................. 709 3. Other Chromatographic Methods................................................. 709 Paper Chromatography........................................................... 709 Column Chromatography......................................................... 712 Gas Chromatography............................................................. 713 References....................................................................... 714 CHAPTER 26 Chromatography of Antibiotics.............................. 717 M. Vondracek Antibiotics Research Institute Roztoky near Prague, Czechoslovakia Translated by M. Hefmansky 1. Introduction...................................................................... 717 2. Detection of Antibiotics on Chromatograms....................................... 718 3. Systematic Analysis of New Antibiotics............................................ 720 4. Classes of Antibiotics............................................................. 723 The Penicillins.................................................................... 723 Basic Water-Soluble Antibiotics................................................... 725 Saturated Macrolide Antibiotics................................................... 726 Unsaturated Macrolide Antibiotics................................................ 727 Polypeptide Antibiotics........................................................... 727 Other Antibiotics................................................................. 728 References ....................................................................... 729 CHAPTER 27 Chromatography of Inorganic Ions........................ 735 Graham Nickless School of Chemistry, The University Bristol, England F. H. Pollard, deceased Formerly, School of Chemistry, The University Bristol, England 1. Introduction *The author gratefully acknowledges the help of his associates. R. G. Anderson, D. N. Dolan, M. Henry, J. D. Murray, and D. B. Thomas in collecting references for this review. x\| CONTENTS 2. Adsorption Chromatography ..................................................... 733 Precipitation ..................................................................... Ion Exchange.:................................................................... Metal Complexes................................................................. 739 Modified Supports................................................................ 739 3. Partition Chromatography........................................................ 74° Cations.......................................................................... 74° General........................................................................ 740 Schemes of Qualitative Analysis................................................. 741 Fractionation of Groups of Metals.............................................. 74_ Separation of Rare Metals and Valency States................................... 744 Anions........................................................................... 743 General........................................................................ 745 Acids of N, P, S, and Halogens.................................................. 745 Quantitative Methods............................................................. 746 Radioactive Elements............................................................. 747 Metal Complexes................................................................. 748 4. Thin-Layer Chromatography...................................................... 749 General.......................................................................... 749 Cations.......................................................................... 750 Anions........................................................................... 752 Nonionic Species................................................................. 753 Ion Exchange and Electrophoresis................................................. 754 References....................................................................... 755 CHAPTER 28 Chromatography of Nonhydrocarbon Gases............... 761 Jaroslav Janak Institute of Instrumental Analytical Chemistry Czechoslovak Academy of Science Brno, Czechoslovakia 1. Introduction..................................................................... 761 2. Separation of Gases and Special Technical Applications............................ 762 Permanent Gases................................................................. 762 Synthesis. Fuel, Town, and Similar Gases........................................ 766 Gases from Solids.............................................................. 767 Elemental Analysis............................................................. 767 Gases from Liquids............................................................. 767 Other Applications............................................................. 768 Other Nonhydrocarbon Gases..................................................... 768 Noble Gases.............................................................. 768 Gaseous Nitrogen Compounds.................................................. 769 Gaseous Sulfur Compounds.................................................... 771 Halogens and Halogen Compounds............................................. 772 Other Inorganic Gases and Vapors............................. 773 3. Trace Analysis............................................. 774 Methods.................................... 774 Concentration of Trace Substances by Sorption and Condensation................ 774 CONTENTS xli Increase of Detection Sensitivity ................................................ 777 Other Methods................................................................. 777 4. Isotope Separation................................................................ 779 Separation of Hydrogen Isotopes and Spin Isomers................................. 779 Separation by Displacement Technique.......................................... 779 Separation by Elution Technique................................................ 781 Separation of Other Isotopes and Isotopic Gaseous Compounds.................... 783 References....................................................................... 785 CHAPTER 29 Chromatography of Hydrocarbons.......................... 794 D. H. Desty British Petroleum Company, Ltd. Sunbury-on-Thames, Middlesex, England A. Gold up British Petroleum Company Ltd. Sunbury-on-Thames, Middlesex, England 1. Introduction...................................................................... 794 2. Gas Chromatography............................................................. 794 Gas Mixtures..................................................................... 795 Janak Method.................................................................. 796 Thermal Conductivity Method.................................................. 798 Light Distillates.................................................................. 800 Separation..................................................................... 801 Identification................................................................... 802 Determination ................................................................. 804 High-Molecular Weight Fractions................................................. 805 Stationary Phases .............................................................. 805 Simple Direct Applications ..................................................... 806 Detailed Composition Studies................................................... 807 Capillary Columns................................................................ 807 Apparatus..................................................................... 808 Column Types ................................................................. 809 Hydrocarbon Separations....................................................... 812 Trace Analysis.................................................................... 817 Process Monitoring and Control .................................................. 817 3. Liquid Chromatography.......................................................... 820 Liquid-Solid Chromatography.................................................... 820 Displacement Method.......................................................... 820 Elution Method................................................................ 821 Liquid-Liquid Chromatography................................................... 824 Industrial Applications ........................................................... 825 References ....................................................................... 826 Index............................................................................ 831
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spelling	Chromatography Erich Heftmann [Ed.] 2. ed. New York Reinhold 1967 XLI, 851 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Reinhold chemistry textbook series Chromatographie (DE-588)4010153-8 gnd rswk-swf Chromatographie (DE-588)4010153-8 s DE-604 Heftmann, Erich Sonstige oth HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010316451&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Chromatography Chromatographie (DE-588)4010153-8 gnd
subject_GND	(DE-588)4010153-8
title	Chromatography
title_auth	Chromatography
title_exact_search	Chromatography
title_full	Chromatography Erich Heftmann [Ed.]
title_fullStr	Chromatography Erich Heftmann [Ed.]
title_full_unstemmed	Chromatography Erich Heftmann [Ed.]
title_short	Chromatography
title_sort	chromatography
topic	Chromatographie (DE-588)4010153-8 gnd
topic_facet	Chromatographie
url	http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010316451&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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