Horst Friebolin

Basic One- and

Two-Dimensional
NMR Spectroscopy

Third Revised Edition

Translated by
Jack K. Becconsall

WILEY-VCH
Weinheim • New York • Chichester • Brisbane • Singapore • Toronto
Contents

1 The Physical Basis of

NMR Spectroscopy 1
1.1 Introduction 1
1.2 Nuclear Angular Momentum and Magnetic
Moment 2
1.3 Nuclei in a Static Magnetic Field 4
1.3.1 Directional Quantization 4
1.3.2 Energy of the Nuclei in the Magnetic Field . . . . 4
1.3.3 Populations of the Energy Levels 6
1.3.4 Macroscopic Magnetization 6
1.4 Basic Principles of the NMR Experiment 7
1.4.1 The Resonance Condition 7
1.4.2 Basic Principle of the NMR Measurement . . . . 8
1.5 The Pulsed NMR Method 9
1.5.1 The Pulse 9
1.5.2 The Pulse Angle 10
1.5.3 Relaxation 13
1.5.4 The Time and Frequency Domains;
the Fourier Transformation 14
1.5.5 Spectrum Accumulation 16
1.5.6 The Pulsed NMR Spectrometer 18
1.6 Spectral Parameters: a Brief Survey 22
1.6.1 The Chemical Shift 22
1.6.1.1 Nuclear Shielding 22
1.6.1.2 Reference Compounds and the <5-Scale 24
1.6.2 Spin-Spin Coupling 26
1.6.2.1 The Indirect Spin-Spin Coupling 26
1.6.2.2 Coupling to One Neighboring Nucleus
(AX Spin System) 27
1.6.2.3 Coupling to Two Equivalent Neighboring Nuclei
(AX2 Spin System) 28
1.6.2.4 Coupling to Three or More Equivalent
Neighboring Nuclei (AXn Spin System) 30
1.6.2.5 Multiplicity Rules 30
1.6.2.6 Couplings between Three Non-equivalent
Nuclei (AMX Spin System) 31
1.6.2.7 Couplings between Equivalent Nuclei
(A n Spin Systems) 32
1.6.2.8 The Order of a Spectrum 33

XV
1.6.2.9 Couplings between Protons and other Nuclei;
13
C Satellite Spectra 33
1.6.3 The Intensities of the Resonance Signals 34
1.6.3.1 J H Signal Intensities 34
1.6.3.2 13C Signal Intensities 35
1.6.4 Summary 37
1.7 "Other" Nuclides 38
1.7.1 Nuclides with I=Yi 39
1.7.2 Nuclides with I>Vi 40
1.8 Bibliography for Chapter 1 41

2 The Chemical Shift 43

2.1 Introduction 43
2.1.1 Influence of the Charge Density on the
Shielding 44
2.1.2 Effects of Neighboring Groups 47
2.1.2.1 Magnetic Anisotropy of Neighboring Groups. . 47
2.1.2.2 Ring Current Effects 49
2.1.2.3 Electric Field Effects 51
2.1.2.4 Intermolecular Interactions - Hydrogen
Bonding and Solvent Effects 51
2.1.2.5 Isotope Effects 51
2.1.3 Summary 52
2.2 'H Chemical Shifts of Organic Compounds . . . 53
2.2.1 Alkanes and Cycloalkanes 54
2.2.2 Alkenes 56
2.2.3 Arenes 56
2.2.4 Alkynes 57
2.2.5 Aldehydes 58
2.2.6 OH, SH, NH 59
I3
2.3 C Chemical Shifts of Organic Compounds . . . 60
2.3.1 Alkanes and Cycloalkanes 61
2.3.2 Alkenes 63
2.3.3 Arenes . 64
2.3.4 Alkynes 66
2.3.5 Allenes 66
2.3.6 Carbonyl and Carboxy Compounds 66
2.3.6.1 Aldehydes and Ketones 67
2.3.6.2 Carboxylic Acids and Derivatives 68
2.4 Relationships between the Spectrum and the
Molecular Structure 70
2.4.1 Equivalence, Symmetry and Chirality 70
2.4.2 Homotopic, Enantiotopic and Diastereotopic
Groups 74
2.4.3 Summary 77
2.5 Chemical Shifts of "Other" Nuclidess 78
2.6 Bibliography for Chapter 2 83

XVIII
3 Indirect Spin-Spin Coupling 85
3.1 Introduction 85
3.2 H,H Coupling Constants and Chemical
Structure 87
3.2.1 Geminal Couplings 2 /(H,H) 87
3.2.1.1 Dependence on Bond Angle 87
3.2.1.2 Substituent Effects 87
3.2.1.3 Effects of Neighboring 7T-Electrons 87
3.2.2 Vicinal Couplings 3 /(H,H) 89
3.2.2.1 Dependence on the Dihedral Angle 90
3.2.2.2 Substituent Effects 93
3.2.3 H,H Couplings in Aromatic Compounds 95
3.2.4 Long-range Couplings 96
3.3 C,H Coupling Constants and Chemical
Structure 97
3.3.1 C,H Couplings through One Bond V(C,H) . . . 97
3.3.1.1 Dependence on the s-Fraction 97
3.3.1.2 Substituent Effects 97
3.3.2 C,H Couplings through Two or More Bonds . . . 98
3.3.2.1 Geminal Couplings (i. e. 27(C,H) in H-C-13C) . . 98
3.3.2.2 Vicinal Couplings (i. e. 37(C,H) in H-C-C-13C) . 99
3.3.2.3 Long-range Couplings 3+ "/(C,H) 99
3.3.3 C,H Couplings in Benzene Derivatives . 99
3.4 C,C Coupling Constants and Chemical
Structure 100
3.5 Correlations between C,H and H,H Coupling
Constants 101
3.6 Coupling Mechanisms 102
3.6.1 The Electron-Nuclear Interaction 102
3.6.2 H,D Couplings 104
3.6.3 Relationship between the Coupling and the
Lifetime of a Spin State 105
3.6.4 Couplings through Space 106
3.7 Couplings of "Other" Nuclides
(Heteronuclear Couplings) 106
3.8 Bibliography for Chapter 3 109

4 Spectrum Analysis and Calculations Ill

4.1 Introduction Ill
4.2 Nomenclature 113
4.2.1 Systematic Notation for Spin Systems 113
4.2.2 Chemical and Magnetic Equivalence 114
4.3 Two-Spin Systems 116
4.3.1 The AX Spin System 116
4.3.2 The AB Spin System 118
4.4 Three-Spin Systems 120
4.4.1 The AX2, AK2, AB 2 , and A 3 Spin Systems . . . 120
4.4.2 The AMX and ABX Spin Systems 121
4.5 Four-Spin Systems 123
4.5.1 A2X2 and A 2 B 2 Spin Systems 123
4.5.2 The AA'XX' and AA'BB' Spin Systems 124
4.6 Spectrum Simulation and Iteration 125
4.7 Analysis of "C NMR Spectra 126
4.8 Bibliography for Chapter 4 127

5 Double Resonance Experiments 129

5.1 Introduction 129
5.2 Spin Decoupling in 'H NMR Spectroscopy . . . . 130
5.2.1 Simplification of Spectra by Selective Spin
Decoupling 130
5.2.2 Suppression of a Solvent Signal 132
5.3 Spin Decoupling in ° C NMR Spectroscopy . . . 133
5.3.1 !H Broad-band Decoupling 133
5.3.2 The Gated Decoupling Experiment 135
X
5.3.3 H Off-Resonance Decoupling 136
5.3.4 Selective Decoupling in 13C NMR Spectroscopy 137
5.4 Bibliography for Chapter 5 138

6 Assignment of lH and 13 C Signals 139

6.1 Introduction 139
6.2 m NMR Spectroscopy 140
6.2.1 Defining the Problem 140
6.2.2 Empirical Correlations for Predicting Chemical
Shifts . . • 141
6.2.2.1 Alkanes (Shoolery's Rule) 141
6.2.2.2 Alkenes 142
6.2.2.3 Benzene Derivatives 143
6.2.3 Decoupling Experiments 145
6.2.4 Effects of Solvent and Temperature . 145
6.2.5 Altering the Chemical Structure of the Sample . 146
D
6.3 C NMR Spectroscopy 147
6.3.1 Defining the Problem . 147
6.3.2 Empirical Correlations for Predicting
Approximate Chemical Shifts 148
6.3.2.1 Alkanes 148
6.3.2.2 Alkenes x 151
6.3.2.3 Benzene Derivatives . 153
6.3.3 Decoupling Experiments 154
6.3.4 T\ Measurements 154
6.3.5 Solvent and Temperature Effects and Shift
Reagents 154
6.3.6 Chemical Changes to the Sample 154
6.4 Computer-aided Assignment of
U
C NMR Spectra 156
6.4.1 Searching for Identical or Related Compounds . 156
6.4.2 Spectrum Prediction 157
6.5 Bibliography for Chapter 6 159
7 Relaxation 161
7.1 Introduction 161
7.2 Spin-Lattice Relaxation of 13C Nuclei (7\) . . . . 162
7.2.1 Relaxation Mechanisms 162
7.2.2 Experimental Determination of 7\; the
Inversion Recovery Experiment 164
7.2.3 Relationships between T\ and Chemical
Structure 168
7.2.3.1 Influence of Protons in CH, CH 2 and CH 3
Groups 168
7.2.3.2 Influence of Molecular Size 169
7.2.3.3 Segmental Mobilities 170
7.2.3.4 Anisotropy of the Molecular Mobility 170
7.2.4 Suppression of the Water Signal 171
7.3 Spin-Spin Relaxation (T2) 171
7.3.1 Relaxation Mechanisms 171
7.3.2 Experimental Determination of T2;
the Spin-Echo Experiment 173
7.3.3 Line-widths of NMR Signals 177
7.4 Bibliography for Chapter 7 178

8 One-Dimensional NMR Experiments

using Complex Pulse Sequences 181
8.1 Introduction 181
8.2 Basic Techniques using Pulse Sequences and
Pulsed Field Gradients 182
8.2.1 The Effect of the Pulse on the Longitudinal
Magnetization (Mz) 183
8.2.2 The Effect of the Pulse on the Transverse
Magnetization Components (Mx- My) 184
8.2.3 The Effect of Pulsed Field Gradients on the
Transverse Magnetization 187
8.3 The /-Modulated Spin-Echo Experiment 192
8.4 The Pulsed Gradient Spin-Echo Experiment . . 200
8.5 Signal Enhancement by Polarization Transfer . . 202
8.5.1 The SPI Experiment 202
8.5.2 The INEPT Experiment 205
8.5.3 The Reverse INEPT Experiment with
Proton Detection 213
8.6 The DEPT Experiment 218
8.7 The Selective TOCSY Experiment 223
8.8 The One-Dimensional INADEQUATE
Experiment 225
8.9 Bibliography for Chapter 8 229
9 Two-Dimensional NMR
Spectroscopy 231
9.1 Introduction 231
9.2 The Two-Dimensional NMR Experiment 232
9.2.1 Preparation, Evolution and Mixing, Data
Acquisition 232
9.2.2 Graphical Representation 236
9.3 Two-Dimensional /-Resolved NMR
Spectroscopy 237
9.3.1 Heteronuclear Two-Dimensional /-Resolved
NMR Spectroscopy 237
9.3.2 Homonuclear Two-Dimensional /-Resolved
NMR Spectroscopy 241
9.4 Two-Dimensional Correlated NMR
Spectroscopy 246
9.4.1 Two-Dimensional Heteronuclear
(C,H)-Correlated NMR Spectroscopy
(HETCOR or C,H-COSY) 247
9.4.2 Two-Dimensional Homonuclear
(H,H)-Correlated NMR Spectroscopy
(H,H-COSY; Long-Range COSY) 255
9.4.3 Reverse Two-Dimensional Heteronuclear
(H.C)-Correlated NMR Spectroscopy
(HSQC; HMQC) 263
9.4.4 The Gradient-Selected (gs-)HMBC Experiment 269
9.4.5 The TOCSY Experiment 273
9.4.6 Two-Dimensional Exchange NMR Spectro-
scopy: the NOESY and EXSY Experiments . . . 276
9.5 The Two-Dimensional INADEQUATE
Experiment 279
9.6 Summary of Chapters 8 and 9 284
9.7 Bibliography for Chapter 9 285

10 The Nuclear Overhauser Effect 287

10.1 Introduction 287
10.2 Theoretical Background 288
10.2.1 The Two-Spin System 288
10.2.2 Enhancement Factors 291
10.2.3 Multi-Spin Systems 292
10.2.4 From the One-Dimensional to the
Two-Dimensional Experiment 293
10.3 Experimental Aspects 294
10.4 Applications 295
10.5 Bibliography for Chapter 10 300

11 Dynamic NMR Spectroscopy (DNMR) .301

11.1 Introduction 301
11.2 Quantitative Calculations 305
11.2.1 Complete Line-shape Analysis 305
11.2.2 The Coalescence Temperature 7c and
the Corresponding Rate Constant kc 307
11.2.3 Activation Parameters 308
11.2.3.1 The Arrhenius Activation Energy EA 308
11.2.3.2 The Free Enthalpy of Activation AGX 309
11.2.3.3 Estimating the Limits of Error 310
11.2.4 Rate Constants in Reactions with Intermediate
Stages 311
11.2.5 Intermolecular Exchange Processes 312
11.3 Applications 313
11.3.1 Rotation about CC Single Bonds 313
11.3.1.1 C(sp3)-C(sp3) Bonds 314
11.3.1.2 C(sp2)-C(sp3) Bonds 314
11.3.1.3 C(sp2)-C(sp2) Bonds 315
11.3.2 Rotation about a Partial Double Bond 315
11.3.3 Inversion at Nitrogen and Phosphorus Atoms . . 317
11.3.4 Ring Inversion 318
11.3.5 Valence Tautomerism 321
11.3.6 Keto-Enol Tautomerism 322
11.3.7 Intermolecular Proton Exchange 323
11.3.8 Reactions and Equilibration Processes 325
11.4 Bibliography for Chapter 11 328

12 Shift Reagents 331

12.1 Lanthanide Shift Reagents (LSRs) 331
12.1.1 Fundamentals 331
12.1.2 Applications and Quantitative Interpretation . . 333
12.2 Chiral Lanthanide Shift Reagents 336
12.3 Chiral Solvents 338
12.4 Bibliography for Chapter 12 341

13 Macromolecules 343
13.1 Introduction 343
13.2 Synthetic Polymers 343
13.2.1 The Tacticity of Polymers * 343
13.2.2 Polymerization of Dienes 347
13.2.3 Copolymers 348
13.3 Bibliography for Chapter 13 350

14 NMR Spectroscopy in Biochemistry

and Medicine 351
14.1 Introduction 351
14.2 Elucidating Reaction Pathways in Biochemistry 352
14.2.1 Syntheses using Singly 13C-Labeled Precursors . 352
14.2.1.1 Low Levels of 13C Enrichment 352
14.2.1.2 High Levels of 13C Enrichment 354
14.2.2 Syntheses using Doubly 13C-Labeled Precursors 355
14.3 High Resolution in vivo NMR Spectroscopy . . . 357
14.3.1 The Problem and its Solution 357
31
14.3.2 P NMR Experiments 358
14.3.3 m and 13C NMR Experiments 361
14.4 Magnetic Resonance Tomography 362
14.4.1 Basic Principles and Experimental
Considerations 362
14.4.2 Applications 370
14.5 Bibliography for Chapter 14 372

Subject Index 375

Index of Compounds 383