200 and More NMR Experiments: A Practical CourseISBN: 9783527310678
854 pages
July 2004

Description
· 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/betaSELINCORTOCSY, WET, DOSY, ctCOSY, HMSC, HSQC with adiabatic pulses, HETLOC. Jresolved HMBC, (1,1) and (1,n)ADEQUATE, STD, REDOR, and HRMAS.
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 15Nlabeled human ubiquitin. Techniques used include the constant time principle, the PEP method, filters, gradient selection, and the echo/antiecho 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 workbook 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
Table of Contents
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 ProbeHead 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 PulseDuration 14
Exp. 2.1: Determination of the 90° 1H Transmitter PulseDuration 15
Exp. 2.2: Determination of the 90° 13C Transmitter PulseDuration 18
Exp. 2.3: Determination of the 90° 1H Decoupler PulseDuration 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: ComputerAided 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: LowPower 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 OffResonance Decoupled 13C NMR Spectra 122
Exp. 4.12: The Gated 1HDecoupling Technique 125
Exp. 4.13: The Inverse Gated 1HDecoupling Technique 128
Exp. 4.14: 1H SingleFrequency Decoupling of 13C NMR Spectra 131
Exp. 4.15: 1H LowPower 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: LowTemperature Calibration Using Methanol 141
Exp. 5.2: HighTemperature Calibration Using 1,2Ethanediol 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 RotatingFrame Relaxation Time T1ρ 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: DEPT135 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: 1DINADEQUATE 197
Exp. 6.14: The BIRD Filter 201
Exp. 6.15: TANGO 204
Exp. 6.16: The Heteronuclear DoubleQuantum Filter 207
Exp. 6.17: Purging with a SpinLock Pulse 210
Exp. 6.18: Water Suppression by Presaturation 213
Exp. 6.19: Water Suppression by the JumpandReturn 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 LongRange 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 SolventInduced 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 1HDecoupling 318
Exp. 8.20: NMR Using LiquidCrystal Solvents 321
Chapter 9 Heteronuclear NMR Spectroscopy 324
Exp. 9.1: 1HDecoupled 15N NMR Spectra Using DEPT 330
Exp. 9.2: 1HCoupled 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 SpinLock 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 JResolved 1H NMR Spectroscopy 367
Exp. 10.2: 2D JResolved 13C NMR Spectroscopy 370
Exp. 10.3: The Basic H,HCOSY Experiment 373
Exp. 10.4: LongRange COSY 377
Exp. 10.5: PhaseSensitive COSY 380
Exp. 10.6: PhaseSensitive COSY45 383
Exp. 10.7: E.COSY 386
Exp. 10.8: DoubleQuantumFiltered COSY with Presaturation 389
Exp. 10.9: Fully Coupled C,H Correlation (FUCOUP) 393
Exp. 10.10: C,HCorrelation by Polarization Transfer (HETCOR) 396
Exp. 10.11: LongRange C,HCorrelation by Polarization Transfer 399
Exp. 10.12: C,H Correlation via LongRange Couplings (COLOC) 402
Exp. 10.13: The Basic HMQC Experiment 405
Exp. 10.14: PhaseSensitive HMQC with BIRD Filter and GARP Decoupling 409
Exp. 10.15: Poor Man’s Gradient HMQC 412
Exp. 10.16: PhaseSensitive 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: 2DINADEQUATE 441
Exp. 10.24: The EXSY Experiment 445
Exp. 10.25: X,YCorrelation 448
Chapter 11 1D NMR Spectroscopy with Pulsed Field Gradients 453
Exp. 11.1: Calibration of Pulsed Field Gradients 455
Exp. 11.2: Gradient Preemphasis 458
Exp. 11.3: Gradient Amplifier Test 461
Exp. 11.4: Determination of Pulsed Field Gradient RingDown Delays 464
Exp. 11.5: The Pulsed Field Gradient SpinEcho Experiment 467
Exp. 11.6: Excitation Pattern of Selective Pulses 470
Exp. 11.7: The Gradient Heteronuclear DoubleQuantum Filter 474
Exp. 11.8: The Gradient zzFilter 477
Exp. 11.9: The GradientSelected Dual Step LowPass Filter 480
Exp. 11.10: gsSELCOSY 484
Exp. 11.11: gsSELTOCSY 488
Exp. 11.12: DPFGSENOE 492
Exp. 11.13: gsSELINCOR 496
Exp. 11.14: α/βSELINCORTOCSY 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: INEPTDOSY 518
Exp. 11.21: DOSYHMQC 521
Chapter 12 2D NMR Spectroscopy With Field Gradients 525
Exp. 12.1: gsCOSY 526
Exp. 12.2: ConstantTime COSY 530
Exp. 12.3: PhaseSensitive gsDQFCOSY 534
Exp. 12.4: gsHMQC 538
Exp. 12.5: gsHMBC 542
Exp. 12.6: ACCORDHMBC 546
Exp. 12.7: HMSC 550
Exp. 12.8: PhaseSensititive gsHSQC with Sensitivity Enhancement 554
Exp. 12.9: Edited HSQC with Sensitivity Enhancement 558
Exp. 12.10: HSQC with Adiabatic Pulses for HighField Instruments 563
Exp. 12.11: gsTOCSY 567
Exp. 12.12: gsHMQCTOCSY 571
Exp. 12.13: gsHETLOC 575
Exp. 12.14: gsJResolved HMBC 581
Exp. 12.15: 2QHMBC 585
Exp. 12.16: 1HDetected 2D INEPTINADEQUATE 589
Exp. 12.17: 1,1ADEQUATE 593
Exp. 12.18: 1,nADEQUATE 597
Exp. 12.19: gsNOESY 601
Exp. 12.20: gsHSQCNOESY 604
Exp. 12.21: gsHOESY 608
Exp. 12.22: 1H,15N Correlation with gsHMQC 612
Chapter 13 The Third Dimension 616
Exp. 13.1: 3D HMQCCOSY 618
Exp. 13.2: 3D gsHSQCTOCSY 622
Exp. 13.3: 3D H,C,PCorrelation 626
Exp. 13.4: 3D HMBC 630
Chapter 14 SolidState NMR Spectroscopy 634
Exp. 14.1: Shimming SolidState ProbeHeads 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: HighResolution MagicAngle Spinning 663
Chapter 15 Protein NMR 666
Exp. 15.1: Pulse Determination for Protein NMR 670
Exp. 15.2: HNHSQC 673
Exp. 15.3: HCHSQC 678
Exp. 15.4: MUSIC 682
Exp. 15.5: HNCorrelation using TROSY 688
Exp. 15.6: HNTOCSYHSQC 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: HCCHTOCSY 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: HNNOESYHSQC 766
Exp. 15.18: HCNOESYHSQC 773
Exp. 15.19: 3D HCNNOESY 779
Exp. 15.20: HNCAJ 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 SpinCoupling Data for Ethyl Crotonate and Strychnine
Glossary and Index 804
New to This Edition
 24 new experiments have been inserted into the 14 chapters that were in the 2nd edition, e.g., α/βSELINCORTOCSY, WET, DOSY, ctCOSY, HMSC, HSQC with adiabatic pulses, HETLOC. Jresolved HMBC, (1,1) and (1,n)ADEQUATE, STD, REDOR, and HRMAS.
 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 15Nlabeled human ubiquitin.
 Techniques used include the constant time principle, the PEP method, filters, gradient selection, and the echo/antiecho procedure.
The Wiley Advantage
 This workbook will guide you safely, in stepbystep
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?
 The guide has been written by experts in this field, following the principle of learning by doing.
 All experiments have been specially performed for this book, exactly as described and depicted, exactly as described and shown in the spectra that are reproduced.
 Being a reference source and workbook forthe 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.
Reviews
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 HNMR 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 1D trials to more complex 3D experiments on proteins."
Analytical Chemistry, November 1, 2004
"The handbook is written by experts and gives very detailed stepbystep 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 liquidstate 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
"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 HNMR 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