200 and More NMR Experiments: A Practical Course

ISBN-13: 9783527310678 / Angielski / Miękka / 2004 / 854 str.

Stefan Berger; Siegmar Braun

Opis

This work-book will guide you safely, in step-by-step descriptions, through every detail of the NMR experiments within, beginning with 1D routine experiments and ending with a series of advanced 3D experiments on a protein:

• Which experiment can best yield the desired information?
• How must the chosen experiment be performed?
• How does one read the required information from the spectrum?
• How does this particular pulse sequence work?
• Which other experiments give similar information?

This third edition of the book, following its two highly successful predecessors, has been revised and expanded to 206 experiments. They are organized in 15 chapters, covering test procedures and routine spectra, variable temperature measurements, the use of auxiliary reagents, 1D multipulse experiments, spectra of heteronuclides, and the application of selective pulses. The second and third dimensions are introduced using pulsed field gradients, and experiments on solid state materials are described. A key part describes 3D experiments on the protein ubiquitin with 76 amino acids.

What is new in this third edition?

1. 24 new experiments have been inserted into the 14 chapters that were in the 2nd edition, e.g., alpha/beta-SELINCOR-TOCSY, WET, DOSY, ct-COSY, HMSC, HSQC with adiabatic pulses, HETLOC. J-resolved HMBC, (1,1)- and (1, n)-ADEQUATE, STD, REDOR, and HR-MAS.
2. 20 new protein NMR experiments have been specially devised and are collected in the newly added Chapter 15, ProteinNMR, for which one needs a special model sample: fully 13C- and 15N-labeled human ubiquitin. Techniques used include the constanttime principle, the PEP method, filters, gradient selection, and the echo/anti-echo procedure.

The guide has been written by experts in this field, following the principle of learning by doing: all the experiments have been specially performed for this book, exactly as described and shown in the spectra that are reproduced. Being a reference source and work-book for the NMR laboratory as well as a textbook, it is a must for every scientist working with NMR, as well as for students preparing for their laboratory courses.

Recenzje

"This book is an excellent catalogue of useful NMR experiments for people who are looking for the most suitable experiment to solve a specific problem.
It collects in one place all the currently pulse sequences from liquid NMR spectroscopy, discusses their relative merits, the time required to perform them and gives experimental examples measured by the authors for this book. ... In conclusion, I think this book is a great encyclopedia of the techniques of modern liquid state NMR spectroscopy. It is highly readabele and should be on the shelf of any serious NMR spectroscopist, who does more complicated experiments than routine H–NMR spectroscopy. Finally instrument vendors should consider packing at least one copy of this book with every new NMR machine and using it as an educational toot when installing the machine."

Dr. Gerd Buntkowsky, FSU Jena, Zeitschrift für Physikalische Chemie, Band 218, Heft 11


"This third edition serves as a detailed guide to NMR, complete with 206 experiments ranging form 1–D trials to more complex 3–D experiments on proteins."

Analytical Chemistry, November 1, 2004

"The handbook is written by experts and gives very detailed step–by–step instructions. ... This excellent book is very well written and builds on the success of the earlier version that was largely due to its clarity, information content and the fact that the methods worked."
J. Lindon, Chromatographia 2005, Vol. 61/No. 1/2


"I highly recommend this book to all scientists who are trying to implement new experimental schemes in liquid–state NMR spectroscopy. It is a very useful NMR >cookbook< and a good starting point to find additional detailed information about experimental methods."

Dr. Matthias Ernst, ETH Zürich, Physical Chemistry, ChemPhysChem, 5/2005


"...I find it to be one of the most useful books on my shelf...each new edition has brought substantial improvements...you will not be sorry if you acquire a copy for your personal library."
Applied Spectroscopy, May 2005

Spis treści

Preface v

Chapter 1 The NMR Spectrometer 1

1.1 Components of an NMR Spectrometer 1

1.1.1 The Magnet 1

1.1.2 The Spectrometer Cabinet 2

1.1.3 The Computer 3

1.1.4 Maintenance 3

1.2 Tuning a Probe–Head 3

1.3 The Lock Channel 4

1.4 The Art of Shimming 6

1.4.1 The Shim Gradients 6

1.4.2 The Shimming Procedure 8

1.4.3 Gradient Shimming 11

Chapter 2 Determination of Pulse–Duration 14

Exp. 2.1: Determination of the 90° 1H Transmitter Pulse–Duration 15

Exp. 2.2: Determination of the 90° 13C Transmitter Pulse–Duration 18

Exp. 2.3: Determination of the 90° 1H Decoupler Pulse–Duration 21

Exp. 2.4: The 90° 1H Pulse with Inverse Spectrometer Configuration 24

Exp. 2.5: The 90° 13C Decoupler Pulse with Inverse Configuration 27

Exp. 2.6: Composite Pulses 30

Exp. 2.7: Radiation Damping 33

Exp. 2.8: Pulse and Receiver Phases 36

Exp. 2.9: Determination of Radiofrequency Power 39

Chapter 3 Routine NMR Spectroscopy and Standard Tests 43

Exp. 3.1: The Standard 1H NMR Experiment 44

Exp. 3.2: The Standard 13C NMR Experiment 49

Exp. 3.3: The Application of Window Functions 54

Exp. 3.4: Computer–Aided Spectral Analysis 58

Exp. 3.5: Line Shape Test for 1H NMR Spectroscopy 61

Exp. 3.6: Resolution Test for 1H NMR Spectroscopy 64

Exp. 3.7: Sensitivity Test for 1H NMR Spectroscopy 67

Exp. 3.8: Line Shape Test for 13C NMR Spectroscopy 70

Exp. 3.9: ASTM Sensitivity Test for 13C NMR Spectroscopy 73

Exp. 3.10: Sensitivity Test for 13C NMR Spectroscopy 76

Exp. 3.11: Quadrature Image Test 79

Exp. 3.12: Dynamic Range Test for Signal Amplitudes 82

Exp. 3.13: 13° Phase Stability Test 85

Exp. 3.14: Radiofrequency Field Homogeneity 88

Chapter 4 Decoupling Techniques 91

Exp. 4.1: Decoupler Calibration for Homonuclear Decoupling 92

Exp. 4.2: Decoupler Calibration for Heteronuclear Decoupling 95

Exp. 4.3: Low–Power Calibration for Heteronuclear Decoupling 98

Exp. 4.4: Homonuclear Decoupling 101

Exp. 4.5: Homonuclear Decoupling at Two Frequencies 104

Exp. 4.6: The Homonuclear SPT Experiment 107

Exp. 4.7: The Heteronuclear SPT Experiment 110

Exp. 4.8: The Basic Homonuclear NOE Difference Experiment 113

Exp. 4.9: 1D Nuclear Overhauser Difference Spectroscopy 116

Exp. 4.10: 1D NOE Spectroscopy with Multiple Selective Irradiation 119

Exp. 4.11: 1H Off–Resonance Decoupled 13C NMR Spectra 122

Exp. 4.12: The Gated 1H–Decoupling Technique 125

Exp. 4.13: The Inverse Gated 1H–Decoupling Technique 128

Exp. 4.14: 1H Single–Frequency Decoupling of 13C NMR Spectra 131

Exp. 4.15: 1H Low–Power Decoupling of 13C NMR Spectra 134

Exp. 4.16: Measurement of the Heteronuclear Overhauser Effect 137

Chapter 5 Dynamic NMR Spectroscopy 140

Exp. 5.1: Low–Temperature Calibration Using Methanol 141

Exp. 5.2: High–Temperature Calibration Using 1,2–Ethanediol 145

Exp. 5.3: Dynamic 1H NMR Spectroscopy on Dimethylformamide 149

Exp. 5.4: The Saturation Transfer Experiment 152

Exp. 5.5: Measurement of the Rotating–Frame Relaxation Time T1 155

Chapter 6 1D Multipulse Sequences 159

Exp. 6.1: Measurement of the Spin Lattice Relaxation Time T1 160

Exp. 6.2: Measurement of the Spin Spin Relaxation Time T2 164

Exp. 6.3: 13C NMR Spectra with SEFT 167

Exp. 6.4: 13C NMR Spectra with APT 170

Exp. 6.5: The Basic INEPT Technique 173

Exp. 6.6: INEPT+ 176

Exp. 6.7: Refocused INEPT 179

Exp. 6.8: Reverse INEPT 182

Exp. 6.9: DEPT–135 185

Exp. 6.10: Editing 13C NMR Spectra Using DEPT 188

Exp. 6.11: DEPTQ 191

Exp. 6.12: Multiplicity Determination Using PENDANT 194

Exp. 6.13: 1D–INADEQUATE 197

Exp. 6.14: The BIRD Filter 201

Exp. 6.15: TANGO 204

Exp. 6.16: The Heteronuclear Double–Quantum Filter 207

Exp. 6.17: Purging with a Spin–Lock Pulse 210

Exp. 6.18: Water Suppression by Presaturation 213

Exp. 6.19: Water Suppression by the Jump–and–Return Method 216

Chapter 7 NMR Spectroscopy with Selective Pulses 219

Exp. 7.1: Determination of a Shaped 90° 1H Transmitter Pulse 220

Exp. 7.2: Determination of a Shaped 90° 1H Decoupler Pulse 223

Exp. 7.3: Determination of a Shaped 90° 13C Decoupler Pulse 226

Exp. 7.4: Selective Excitation Using DANTE 229

Exp. 7.5: SELCOSY 232

Exp. 7.6: SELINCOR: Selective Inverse H,C Correlation via 1J(C,H) 235

Exp. 7.7: SELINQUATE 238

Exp. 7.8: Selective TOCSY 242

Exp. 7.9: INAPT 246

Exp. 7.10: Determination of Long–Range C,H Coupling Constants 249

Exp. 7.11: SELRESOLV 252

Exp. 7.12: SERF 255

Chapter 8 Auxiliary Reagents, Quantitative Determinations, and Reaction Mechanisms 258

Exp. 8.1: Signal Separation Using a Lanthanide Shift Reagent 259

Exp. 8.2: Signal Separation of Enantiomers Using a Chiral Shift Reagent 262

Exp. 8.3: Signal Separation of Enantiomers Using a Chiral Solvating Agent 265

Exp. 8.4: Determination of Enantiomeric Purity with Pirkle s Reagent 268

Exp. 8.5: Determination of Enantiomeric Purity by 31P NMR 271

Exp. 8.6: Determination of Absolute Configuration by the Advanced Mosher Method 274

Exp. 8.7: Aromatic Solvent–Induced Shift (ASIS) 277

Exp. 8.8: NMR Spectroscopy of OH Protons and H/D Exchange 280

Exp. 8.9: Water Suppression Using an Exchange Reagent 283

Exp. 8.10: Isotope Effects on Chemical Shielding 286

Exp. 8.11: pKa Determination by 13C NMR 290

Exp. 8.12: Determination of Association Constants Ka 293

Exp. 8.13: Saturation Transfer Difference NMR 298

Exp. 8.14: The Relaxation Reagent Cr(acac)3 302

Exp. 8.15: Determination of Paramagnetic Susceptibility by NMR 305

Exp. 8.16: 1H and 13C NMR of Paramagnetic Compounds 308

Exp. 8.17: The CIDNP Effect 312

Exp. 8.18: Quantitative 1H NMR Spectroscopy: Determination of the Alcohol Content of Polish Vodka 315

Exp. 8.19: Quantitative 13C NMR Spectroscopy with Inverse Gated 1H–Decoupling 318

Exp. 8.20: NMR Using Liquid–Crystal Solvents 321

Chapter 9 Heteronuclear NMR Spectroscopy 324

Exp. 9.1: 1H–Decoupled 15N NMR Spectra Using DEPT 330

Exp. 9.2: 1H–Coupled 15N NMR Spectra Using DEPT 333

Exp. 9.3: 19F NMR Spectroscopy 336

Exp. 9.4: 29Si NMR Spectroscopy Using DEPT 339

Exp. 9.5: 29Si NMR Spectroscopy Using Spin–Lock Polarization 342

Exp. 9.6: 119Sn NMR Spectroscopy 346

Exp. 9.7: 2H NMR Spectroscopy 349

Exp. 9.8: 11B NMR Spectroscopy 352

Exp. 9.9: 17O NMR Spectroscopy Using RIDE 355

Exp. 9.10: 47/49Ti NMR Spectroscopy Using ARING 358

Chapter 10 The Second Dimension 362

Exp. 10.1: 2D J–Resolved 1H NMR Spectroscopy 367

Exp. 10.2: 2D J–Resolved 13C NMR Spectroscopy 370

Exp. 10.3: The Basic H,H–COSY Experiment 373

Exp. 10.4: Long–Range COSY 377

Exp. 10.5: Phase–Sensitive COSY 380

Exp. 10.6: Phase–Sensitive COSY–45 383

Exp. 10.7: E.COSY 386

Exp. 10.8: Double–Quantum–Filtered COSY with Presaturation 389

Exp. 10.9: Fully Coupled C,H Correlation (FUCOUP) 393

Exp. 10.10: C,H–Correlation by Polarization Transfer (HETCOR) 396

Exp. 10.11: Long–Range C,H–Correlation by Polarization Transfer 399

Exp. 10.12: C,H Correlation via Long–Range Couplings (COLOC) 402

Exp. 10.13: The Basic HMQC Experiment 405

Exp. 10.14: Phase–Sensitive HMQC with BIRD Filter and GARP Decoupling 409

Exp. 10.15: Poor Man s Gradient HMQC 412

Exp. 10.16: Phase–Sensitive HMBC with BIRD Filter 415

Exp. 10.17: The Basic HSQC Experiment 418

Exp. 10.18: The HOHAHA or TOCSY Experiment 422

Exp. 10.19: HETLOC 426

Exp. 10.20: The NOESY Experiment 430

Exp. 10.21: The CAMELSPIN or ROESY Experiment 434

Exp. 10.22: The HOESY Experiment 438

Exp. 10.23: 2D–INADEQUATE 441

Exp. 10.24: The EXSY Experiment 445

Exp. 10.25: X,Y–Correlation 448

Chapter 11 1D NMR Spectroscopy with Pulsed Field Gradients 453

Exp. 11.1: Calibration of Pulsed Field Gradients 455

Exp. 11.2: Gradient Pre–emphasis 458

Exp. 11.3: Gradient Amplifier Test 461

Exp. 11.4: Determination of Pulsed Field Gradient Ring–Down Delays 464

Exp. 11.5: The Pulsed Field Gradient Spin–Echo Experiment 467

Exp. 11.6: Excitation Pattern of Selective Pulses 470

Exp. 11.7: The Gradient Heteronuclear Double–Quantum Filter 474

Exp. 11.8: The Gradient zz–Filter 477

Exp. 11.9: The Gradient–Selected Dual Step Low–Pass Filter 480

Exp. 11.10: gs–SELCOSY 484

Exp. 11.11: gs–SELTOCSY 488

Exp. 11.12: DPFGSE–NOE 492

Exp. 11.13: gs–SELINCOR 496

Exp. 11.14: / –SELINCOR–TOCSY 499

Exp. 11.15: GRECCO 503

Exp. 11.16: WATERGATE 506

Exp. 11.17: Water Suppression by Excitation Sculpting 509

Exp. 11.18: Solvent Suppression Using WET 512

Exp. 11.19: DOSY 515

Exp. 11.20: INEPT–DOSY 518

Exp. 11.21: DOSY–HMQC 521

Chapter 12 2D NMR Spectroscopy With Field Gradients 525

Exp. 12.1: gs–COSY 526

Exp. 12.2: Constant–Time COSY 530

Exp. 12.3: Phase–Sensitive gs–DQF–COSY 534

Exp. 12.4: gs–HMQC 538

Exp. 12.5: gs–HMBC 542

Exp. 12.6: ACCORD–HMBC 546

Exp. 12.7: HMSC 550

Exp. 12.8: Phase–Sensititive gs–HSQC with Sensitivity Enhancement 554

Exp. 12.9: Edited HSQC with Sensitivity Enhancement 558

Exp. 12.10: HSQC with Adiabatic Pulses for High–Field Instruments 563

Exp. 12.11: gs–TOCSY 567

Exp. 12.12: gs–HMQC–TOCSY 571

Exp. 12.13: gs–HETLOC 575

Exp. 12.14: gs–J–Resolved HMBC 581

Exp. 12.15: 2Q–HMBC 585

Exp. 12.16: 1H–Detected 2D INEPT–INADEQUATE 589

Exp. 12.17: 1,1–ADEQUATE 593

Exp. 12.18: 1,n–ADEQUATE 597

Exp. 12.19: gs–NOESY 601

Exp. 12.20: gs–HSQC–NOESY 604

Exp. 12.21: gs–HOESY 608

Exp. 12.22: 1H,15N Correlation with gs–HMQC 612

Chapter 13 The Third Dimension 616

Exp. 13.1: 3D HMQC–COSY 618

Exp. 13.2: 3D gs–HSQC–TOCSY 622

Exp. 13.3: 3D H,C,P–Correlation 626

Exp. 13.4: 3D HMBC 630

Chapter 14 Solid–State NMR Spectroscopy 634

Exp. 14.1: Shimming Solid–State Probe–Heads 635

Exp. 14.2: Adjusting the Magic Angle 639

Exp. 14.3: Hartmann Hahn Matching 642

Exp. 14.4: The Basic CP/MAS Experiment 645

Exp. 14.5: TOSS 649

Exp. 14.6: SELTICS 653

Exp. 14.7: Connectivity Determination in the Solid State 656

Exp. 14.8: REDOR 659

Exp. 14.9: High–Resolution Magic–Angle Spinning 663

Chapter 15 Protein NMR 666

Exp. 15.1: Pulse Determination for Protein NMR 670

Exp. 15.2: HN–HSQC 673

Exp. 15.3: HC–HSQC 678

Exp. 15.4: MUSIC 682

Exp. 15.5: HN–Correlation using TROSY 688

Exp. 15.6: HN–TOCSY–HSQC 692

Exp. 15.7: HNCA 698

Exp. 15.8: HN(CO)CA 705

Exp. 15.9: HNCO 711

Exp. 15.10: HN(CA)CO 718

Exp. 15.11: HCACO 725

Exp. 15.12: HCCH–TOCSY 732

Exp. 15.13: CBCANH 739

Exp. 15.14: CBCA(CO)NH 746

Exp. 15.15: HBHA(CBCACO)NH 753

Exp. 15.16: HN(CA)NNH 760

Exp. 15.17: HN–NOESY–HSQC 766

Exp. 15.18: HC–NOESY–HSQC 773

Exp. 15.19: 3D HCN–NOESY 779

Exp. 15.20: HNCA–J 785

Appendix 1 791

Pulse Programs

Appendix 2 794

Instrument Dialects

Appendix 3 797

Classification of Experiments

Appendix 4 799

Elementary Product Operator Formalism Rules

Appendix 5 802

Chemical Shift and Spin–Coupling Data for Ethyl Crotonate and Strychnine

Glossary and Index 804