STRUCTURE ELUCIDATION BY NMR IN ORGANIC CHEMISTRYSTRUCTURE ELUCIDATION BY NMR IN ORGANIC CHEMISTRY A Practical Guide Third revised edition EBERHARD BREITMAIER University of Bonn, Germany &) JOHN WILEY & SONS, LTDCopyright © 2002 John Wiley & Sons Lid, The Atriun, Southern Gate, Chichester, West Sussex PO19 880, England Telephone (+44) 1243 779777 ‘Email (for orders and customer service enquiries): es-books(@wiley.co.uke ‘Visit our Home Page on www.wileyeurope.com or www.wiley.com All Rights Reserved. No part of this publication may be reproduced, stored in a retrieval system or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning or dolerwise, except unter the (erms of the Copyright, Designs and Patents Act 1988 or under the terms of a licence issued by the Copyright Licensing Agency Ltd, 90 Tottenham Court Road, London WIT 4LP, UK, without the permission in writing of the Publisher. 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The cover shows the "C NMR spectrum of a- and B-D-xylopyranoce at mutarotational equilibrium (35% 0, 65% B, in douterium oxide, 100 MH, '# broadband decoupling) with the CU INADEQUATE contour plot. An interpretation ofthe plot according to principles described in Section 2.2.7 gives the CC bonds of the two Jno and eonfivns the assigennent of he signals in Table 2.12,CONTENTS PREFACE. PREFACE TO THE FIRST EDITION ‘SYMBOLS AND ABBREVIATIONS 24.2 24.2 243 214 22 224 222 223 224 225 226 227 228 229 23 23.4 232 233 234 235 ‘SHORT INTRODUGTION TO BASIC PRINGIPLES AND METHODS Chetnical shit ‘Spin-spin coupling and coupling constants ‘Signal multiplicity (mulplets) ‘Spectra of frst and higher order Chemical and magnetic equivalence Fourier transform (FT) NMR spectra Spin decoupling Nuclear Overhauser effect Relaxation, relaxation times RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR Functional groups ‘Chemical Shits Deuterium exchange *8¢ Chemical shifts SN Chemical shifts Skeletal structure (atom connectives) HH Muttpicites CH Muttpicites HH Coupling constants CH Coupling constants NH Coupling constants HH COSY (gerninal, vicinal, w-relationships of protons) CC INADEQUATE (CC bonds) ‘Two-dimensional cerbon-proton shift correlation via one-bond CH coupling ‘Two-dimensional carbon-proton shift correlation via long-range CH coupling Relative configuration and conformation HH Coupling constants CH Coupling constants NH Coupling constanls ®C Cheinical shifts NOE Difference spectra " 11 11 12 14 16 16 18 a 26 29 30 3 36 39 42 a7 48 atCONTENTS 236 244 24.2 25 254 25.2 253 254 26 26.2 27 112 18-14 16-20 21-22 23-29 30-31 32-42 43-65 HH NOESY and ROESY ‘Absolute configuration Diastereotopism Chiral shift reagents (ee determination) Intra- and intermolecular interactions Anisotropic effects ing current of aromatic compounds Intra- and intermolecular hydrogen bonding Protonation effects Molecular dynamics (fuxionaity) C Spin-attice relaxation times Summary PROBLEMS ‘Appication of one-dimensional 1H NMR Temperature dependent ‘H and °C NMR spectra Application of one-dimensional 'C NMR spectra CC INADEQUATE experiments ‘Application of one: dimensional ‘H and *°C NMR spectra ‘Application of one-dimensional +H, "™C and 'SN NMR spectra ‘Combined application of one and two-dimensional +H and ™™C NMR experiments Identification and structural elucidation of of natural products by one and two-dimensional *H and SC NMR SOLUTIONS TO PROBLEMS Dimethyl cis-cyclopropane-t,2-dicarboxyiate Ethylacryiale cis-1-Methoxybut-1-en-3-yne trans-3-(N-Methy/pyrrol-2-y))propenal 4,9-Bis(pyrtol-2-y)pyrromethene S-Acetylpyridine Anlhracene-,8-dialdehyde trans-Silbene- “Ds7Hz —/ | | | ois | ws ppm 7.26 6.93 4.33 1.38 00 Figure 4.2. 'H NMR spcotrum of ethy! dichloroacetate (CDCl, 25 *C, 80 MHz). The proton of the CHCie group is less shielded (more stongly desided) in comparison with the protons of the CH, and CH residues 1.3. Signal multiplicity (multiplets) The signal multiplicity in first-order spectra (sce Scction 1.4) is the extent to which an NMR sig= nal is split as a result of spin-spin coupling '*. Signals which show no splitting are denoted as singlets (5). Those with two, thrce, four, five, six or seven lines ate: known as doublets (d), triplets (0), quartets (g, Figs 1.2 and 1.3), quinteds (qui), vextety (9x1) and sepiets (sep), respectively, but only where the: lines ofthe multiplet signal are of equal distance apart, and the one coupling con- stant is therefore shared by them all. Where two or three different coupling constants produce a multiplet, this is referred to as a two- or three-fold multiplet, respectively, e.g. a doublet of doub- lets (dd, Fig, 1.3), or a doublet of doublets of doublets (ddd, Fig, 1.3). If both coupling constants of a doublet of doublets ate sufficiently similar (J; ~ J), the middle signals overlap, thus genera. ting a ‘pseudotripler'('r, Fig, 1.3),14 Spectra of first and higher order The ‘IT NMR spectrum of cthyl dichloroacctate (Fig. 1.2), as an example, displays a triplet for the Ci; group (ovo vicinal H), a quartet for the OCH; group (three vicinal H) and a singlet for the CHCl, fragment (no vieinal I! for coupling). Job Jind A, mt | ol Leh aes J] fal (ul fas) JILIN one coxping const ‘wocowing constants Wosinlar eoupingcosints tee coupng cosas ‘quart ‘ube of doublets pseutotplet ttreeold doublet Figure 1.3. Quartot, doubiot of doubiote, peoudotniplot and thraetolé doublet (doublet of doublets of doublete) 1.4 Spectra of first and higher order First-order spectra (multiplets) are observed when the coupling constant is small compared with the frequency difference of chemical shifts between the coupling nuclei *", This is referred to as an Ay, spin system, where nucleus has the smaller and nucleus X has the considerably larger chemical shift. An AX system (Fig. 1.4) consists of an A doublet and an X doublet with the com mon coupling constant J.y. The chemical shifts are measured from the centres of each doublet to the reference resonance. | | (vx- va) >> doe } | Axeystem — JL vy & Figure 1.4. Two-spin system of ype AX will a chemical shif difference whieh Is latge compared with the cou pling conetants (schematic) n=0Shgit 1 1 outlet 1 1 2 Inplet eee 1 3 Quartet 1 3 3 1 4 Quintet 1 4 6: 4 1 5 Seatot 1 5 0 10 5 1 6 Septet 1 6 15 2 6 6 1 Figure 4.5, Relative intensities of first-order lips (Passel rbangle)4 1._ SHORT INTRODUCTION TO BASIC PRINCIPLES AND METHODS Multiplicity rules apply tor first-order spectra (Ay, systems); When coupled, ny nuclei of an element X with nuclear spin quantum number /y= Ys produce a splitting of the signal into ty + 1 Tines; the relative intensities of the individual lines of a first-order multiplet are given by the coeTTicients of the Pascal triangle (Fig. 1.5). The protons of the ethyl group of ethyl dichloroacc- tate Fig. 1.2) ws examples give vise to an A3X2 system with the eoupling constant 'Jyy= 7 Iz; the A protons (with smaller shifl) are split into # triplet (Iwo vieinal protons X, ny-+ 1 = 3); the X pro- tons appear as quartet because of three vicinal A potas (ny + 1 — 4). In general, for a given number, my, of coupled nuclear spins of spin quantum mumiber fy, the A sigial will he split inte (Qn Iy+ 1) multiplet lines (e.g, Fig. 1.9) Spectra of greater complexity may occur for systems where the coupling constant is of similar magnitude to the chemical shift difference between the coupled nuclei. Such a case is referred t0 as an 4,, system, where mucleas A has the smaller and nucleus B the larger chemical shift. ‘An AB system (Fig. 1.6) consists, for example, of an 4 doublet and a B doublet with the common coupling constant Ja, . where the external signal of both doublets is attenuated and the internal signal is enhanced, This is referred to as an AB effect, a roofing’ symmetric to the centre of the AB system *; ‘roofing’ is frequently observed in proton NMR spectra, even in practically first order spectra (Fig, 1.2, ethyl quartet and triplet). The chemical shifts vg and vg arc displaced from the centres of the two doublets, approaching the frequencies of the: mare intense inner signals, Je] Joe (vo-Vs) ® Jeo 1 tt ABsystem — JING Vo ‘A & & Figure 4.6, Two-spin system of type AB with a small chemical shift ilference compared lo the coupling cam stant (gchematic) 41.5 Chemical and magnetic equivalence Chemical equivalence: stomic nuclei in the same chemical environment are chemically equivalent and thus show the same chemical shift. The 2,2'- and 3,3'protons of a 1,4-disubstituted benzene ring, for example, are chemically equivalent because of molecular symmetry cut wauulig: y= 7B? XM HA Oa OCH on al para couning ; Sur = O51 He Magnetic equivalence: chemically equivalent nuclei are magnetically equivalent if they display the same coupling constants with all other nuclear spins of the molecule ’’, For example, the 2,2'-1.6 Fourier transform (FT) NMR spectra 5 (AA) and 3,3-(4,X) protons of a 1,{-disubstituted benzene ring such as 4-nitroanisole are not magnetically cquivalent, becausc the 2-proton A shows an ortho coupling with the 3 proton X (’/— 7-8 Iz), but displays a different para coupling with the 3'-proton X" (‘J — .5-/ Hz). ‘This 1s there- fone referred to as an AA’XY’ system (cg. Fig. 2.6¢) but not as an A2X> or an (AX)2 system. The 'H NMR spectrum in such a case can never he first-order, and the multiplicity rules do not apply. The methyl protons in ethyl dichlormacetate: (Fig. 1.2), however, are chemically and magnetically equi- valent because: the. °Jj coupling constants depend on the geometric relations with the CIs protons and these average (6 the sume for all CH, protons due to cotation about the CC single bond, they are the 4, part of an A,X system characterising an ethoxy group (CH*,—CH*,—O-) in ‘H NMR. 1.6 Fourier transform (FT) NMR spectra ‘There are two basic techniques for recording high-resolution NMR spectra **, In the older CW’ technique, the frequency or field appropriate for the chemical shift range of the nucleus (usually ‘7 is swept by a continuously increasing (or decreasing) radio-frequency. The duration of the sweep is long, typically 2 He/s, or $00.5 fora sweep of 1000 Hl, corresponding to 10 ppm in 100 MHz proron NMR specira. This monachroratic excitation therefore takes a Tong. time (0 recon In the #7 technique, the whole of the Larmor frequency range of the observed nucleus is excited by a radiofrequency pulse. This catiscs transversc magnctisation to build up in the sample. Once excitation stops, the transverse magnetisation decay's exponentially with the time constant 7; of spin-spin relaxation provided the field is perfectly homogencous. In the case of a one-spin system, the corresponding NMR signal is observed as an exponentially decaying alternating voltage (free induction decay, FID); multi-spin systems produce an interference of several exponentially de- caying allervating, vallages, the: pulse interferogram (Fig, 1.7).The frequency of each alternating, vollage is the difference between the individual Larmor frequency of one specific kind of nucleus and the frequency of the exciting pulse. The Fourier transformation (FT) of the pulse interfero- gram produces the Larmor frequeney spectrum; this is the FT NMR spectrum of the type of nucleus being observed. Fourier transformation involving the calculation of all Larmor frequen- cies contributing to the interferogram is performed with the help of a computer within a time of less than 1 s, Putco intrerogam -FTNMR opoctun Fit) ry) wm || sottenn | —— 8s <0 Figure 1.7. Pulse interferogram and FT "NMR spectrum of glycerol, (OCI h)»C! OD. [Ds0, 25°C. 100 MHz]6 1 SIIORT INTRODUCTION TO BASIC PRINCIPLES AND METHODS The main advantage of the technique is the short time required for the procedure (about | s per interferogram), Within a short time a large number of individual interferograms can be accumula ted, thus averaging out clectronic noise (FID accumulation), and making the F'I' method the pre- ferred approach for less scnsitive NMR probes involving isotopes of low natural abundance (°C, "SN), All of the spectra in this hook with the exception of those in Tigs. 1.8, 2.19 and 2.25 arc P7 NMR spectra 1.7 Spin decoupling Spin decoupling (double resonance)**"* is an NMR technique in which, to take the simplest ex- ample, an AX system, the spliting of the A signal due to J, coupling is removed if the sample is irradiated strong enough by a second radiofrequency which resonates with the Larmor frequency of the X nucleus, The 4 signal then appears as a singlet; at the position of the X signal interference is observed between the X Larmor frequency and the decoupling frequency. If the 4 and X nucle are the same isotope (c.g. protons), this is referred to as selective homonuclear decoupling. If A and Yare different, ¢.g. carbon-13 and protons, then itis referred to as heteronuclear decoupling. Figure 1.8 illustrates homonuclear decoupling experiments with the CH protons of 3-amino- aerolein. These give rise to an AMLY system (Fig. .8a). Decoupling of the aldehyde proton X (Fig, |1.8b) simplifies the NMR spectrum to an AM system (4,7 ~ [2.5 Hz); decoupling of the Mf pro ton (Fig. 1.8e) simplifies to an AX system (’J, = 9 Hz). These experiments reveal the connectivi- tics of the protons within the molecule, 2356 SAminoseroen = Tf a — MA a5 ppm 73 $25 Figure 4.6. tiomonuclear decoupling of the GH protons of S:aminuatiolkn (CD40, 25 70, 8 MHZ). (a) "H NMR spectrum; (b) decoupling at 3, ~ 8.6; (0) dacoupling at 3, = 7.3. At tho positon of the decoupled sianal in (b) and (c) interference beats ave absurd Licoune a Ihe superposition of the twn very sinliae frequencies1.7 Spin decoupling z In "© NMR spectroscopy, three kinds of heteronuclear spin decoupling are used ““, In proton broadband decoupling ot °C NMK spectra, decoupling is carried out unselectively across a fre queney range which encompasses the whole range of the proton shifts. The spectrum then displays up to n singlet signals for the n non-equivalent C atoms of the molecule. Figures 1,9a and b demonstrate the effect of proton broadband decoupling in the \C NMR spec- trum of a mixture of ethanol and hexadeuterioethanol. The CH, and CH; signals of ethanol appear as intense singlets upon proton broadband decoupling while the CD; and CD; resonances of the deuteriated compound still display their septet and quintet fine structure; deuterium nuclei are not affected by ‘H decoupling because their Larmor frequencies are far removed from those of pro- tons: further, the nuclear spin quantum number of deuterium is 1,,~ /, in keeping with the general multiplicity rule (2 ity fx + 1, Section 1.4), triplets, quintets and septets are observed for CD, CD and CDs groups, respectively. The relative intensities in these multiplets do not follow Pascal's triangle (1:1:1 triplet for CD; 1:3:4:3:1 quintet for CD3; 1:3:6:7:6:3:1 septct for CDs). “os. ° Lo Nh _eeeee ieee | deme CH3—CH2—OH €D3—CDy—OD a eee eee Ue UeUeUECeCeeeeeeeeeeeececeeceen: ry} rh on ‘31 ° Figure 1.9. "°C NMR spectra of a mixture of ethancl and hexadeutericethanol [27:75 viv, 25 °C. 20 MHz]. (a) 'H thronthand decoupled; (b) without decoupling, The deuterium isotope effect cin — abo on "C chomical ehis is 1.1 and 0.88 ppm for methy! and methylene carbon nucle, respectively In selective proton decoupling of C NMR spectra, decoupling is performed at the: precession frequency of a specific proton. As a result, a singlet only is ahserved for the alluched C atom, Of resonance conditions apply to the other C atoms. For these the individual lines of the CH multi= plots move closer together, and the relative intensities of the: multiplet lines change from those given by the Pascal triangle; extemal signals are attenuated whereas infesual signals are enhanced,a 1_ SHORT INTRODUCTION TO DASIC PRINCIPLES AND METHODS Selective 'H decoupling of "C NMR spectra was used for assignment of the CH connectivities (CH bonds) before the much more cfficient two-dimensional CH shift correlation experiments (see Section 2.2.8) became routine, Offresonance decoupling of the protons was helpful in determt- uning CH multiplicities before better methods became available (sce Section 2.2.2). In pulsed or gated decompling of protons (broadband decoupling only between FIDs), coupled '°C NMR spec- tra are obtained in which the CH multiplets are enhanced by the nuclear Overhauser effect (NOE, see Section 1.8). This method is used! when CH coupling constants are required for structure ana~ lysis because it enhances the multiplets of carbon muclei attached to protons; the signals of quater- nary carbons two bonds apart from a proton are also significantly enfrmced, Figure 1.10 deen strates this for the carbon nuclei in the 4,6-positions of 2,4,6ctrichloropyrimidine 1ea5Hz CN i § FL 2.4,6-Trenirepyrimicine wh ' a | up? 3 oa? | cote j | ded f) |Banee real haemo 1637 12035 pom ae a b Figure 1.10. "°C NMR spoctra of 2,4,6 trichloropyrimidine [C,D,, 75% viv 25 °C, 20 MH]. (a) "C NMR spec leurn without proton decoupling. (b) NOE enhanced coupled "C NMR spectrum (gated decoupling) 1.8 Nuclear Overhauser effect The nuclear Overhauser effect ** (NOR, also an abbreviation for nuclear Overhauser enhance ment) causes the change in intensity (increase or decrease) during decoupling experiments. ‘he maximum possible NOE in high-resolution NMR of solutions depends on the gyromagnetic ratio of the coupled nuclei. Thus, in the homonuclear case such as proton-proton coupling, thc NOL is much less than 0,5, whereas in the most frequently used heteronuclear example, proton decoupling of °C NMR spectra, it may reach 1.988. Instcad of the expected signal intensity of 1, the net result is to increase the signal intensity threefold (1 | 1.988). In proton broadband and gated decoupling of °C NMR spectra, NOE enhancement of signals by a factor of as much as 2 is rautine, as was shown in Figs 1.9 and 1.10.1.8 Nuclear Overhauser effect Quantitative analysis of mixtures is achicved by evaluating the integral steps of “AW NMR spectra, This is demonstrated in Tig. 1.11 for 2,4-pentanedione (acctylacctone) which occurs as an equili brium mixture of 87 % enol and 13 % diketonc. « 24 Penlnesione (oebcetono} HH | = | i oro nome re waomar = | om any , j E57 7" 5.60 ony ome jus | je Pale aaa wis om | b e0% I coe om 3109 F 97600=(* 1014 26.85, Figure 41.11. NMR analysts of the keto-eno! tautomerism of 24-pontanedione [CLCIs, SU% v/v, 25 “C, 60 MHZ for 'H, 20 MHs for "Cl. (a) 'H NMR spectrum with inteorals result: keto : enol = 19 : 87: (b) HV broadband da- ccaupled "™C NMR specirum; (e) °C NMR spectrum obtained by inverse gated “H decoupiing with integrals [result keto : eno! = 18:85 (= 1)10 1 SIIORT INTRODUCTION TO BASIC PRINCIPLES AND METHODS A similar cvaluation of the "C integrals in 'H broadband decoupled '°C NMR spectra fails in most cases because signal intensities arc influenced by nuclcar Overhauser enhancements and relaxation times and these are usually specific for each individual carbon nucleus within a molecule. As a result, deviations arc large (81-93 % cnol) if the keto-cnol equilibrium of 2,4-pentanedione is, analysed hy means of the integrals in the ‘77 broadband decoupled "C NMR spectrum (Fig. LLIb), Inverse: gated decoupling 7, involving proton broadband decoupling only during the FIDs, helps to solve the problem, ‘This technique: provides ‘H broadband decoupled "C NMR spectra with suppressed nuclear Overhauser effect su that signal intensities cau be compared snd Keto-enol tautomerism of 2,4-pentanedione, for example, is unalysed more precisely ay shown in Fig. L.t1e. 1.9 Relaxation, relaxation times Relaxation *** refers to all processes which regenerate the Boltzmann distribution of nuclear spins on their precession states and the resulting equilibrium magnetisation along the static magnetic field. Relaxation also destroys the transverse magnetisation arising from phase coherence of nuclear spins built up upon NMR excitation, Spin-lattice relaxation is the steady (exponential) build-up or regeneration of the: Boltzanann dis- tribution (equilibrium magnetisation) of nuclear spins in the static magnetic fickd. The lattice is the molecular environment of the nuclear spin with which energy is exchanged. The spin-lattice relaxation time, 7). is the time constant for spin-lattice relaxation which is spoci~ fic for every nuclear spin. In PT NMR spectroscopy the spin-lattice relaxation must ‘keep pace’ with the exciting pulses. If the sequence of pulses is too rapid, e.g. faster than 3inax of the 'slo- west’ C atom of a molecule in carbon-13 resonance, a decrease in signal intensity is observed for the ‘slow! C atom due to the spin-lattice relaxation getting ont of step! For this reason, quaternary atoms can be recognised in exrban-13 NMR spectra by their weak signals Spin-spin relaxation is the steady decay of transverse magnetisation (phase coherence of muclear spins) produced by the NMR cacitation where there is perfect homogeneity of the magnetic ficld, It is evident in the shape of the FID (free induction decay), as the exponential decay to zero of the transverse magnetisation produced in the pulsed NMR experiment. The Fourier transformation of the PID signal (time domain) gives the FT NMR spectrum (frequency domain, Fig, 1.7), The spin-spin relaxation time, Ts, is the time constant for spin spin relaxation which is also speci- fic for every nuclear spin (approximately the time constant of FID). For small- to medium-sized ‘molecules in solution 7; ~7;. The value of 1, of a nucleus determines the width of the appropriate NMR signal at half height (half-wideh') according to the uncertainty relationship. The smaller is 1), the broader is the signal. ‘Ihe more rapid is the molecular motion, the larger arc the values of 1 and 1, and the sharper are the signals (‘motional narrowing’). This rule applies to small- and ‘medium-sized molecules of the type most common in organic chemistry. Chemical shifis and coupling constants reveal the static structure of a molecule; relaxation times reflect molecular dynamics,Structure Plucidation Ry NMR bn Organiee Chemisiry: A Practical Guide herhard Brcitmaice 12 Joba Wiley & Sons, Lt. (UB), 0-870-85007-8 (PB) Copyright © 2 ISBNs, 0-470-85006- 2.1 Functional groups it 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 24 Functional groups 21.2 ‘H Chemical Shifts Many functional groups can be identified conclusively hy their 'H chemical shifts“. Important examples are listed in Table 2.1, where the ranges for the prolan shifls are shown in deareasing sequence: aldehydes (5, = 9.5 - 10.5), acetals (6, = 4.5 - 6), alkoxy (dy = 4 - 5.5) and methoxy functions (6, = 3.5 - 4), Nemethyl groups (4, = 3 - 3.5) and methyl residues attached to double bonds such as C=C or C=X (X= N, O, 5) or to aromatic and heteroaromatic skeletons (5y= 1.8 = 25). ‘Table 2.1. '1/ chemical shift ranges for organic compounds Enot Olt 7 Camaxyte asc Phenol OW Alkannl-OH | saci ‘Amide NH ‘Amina-NH Thiol Tiophonol SH Aldenyde-GH Heleroaromatioe Aromatics ‘kena-CH Allyne-CH Acotah Cl? h0- CHiN cH3s- GH at 6-6 or =x Cyvlopropare-CH (Chg Bonded to metal Small shift values for CH or CH protons may indicate cyclopropane units. Proton shifis distin- guish between alkyne CH (generally oy = 2.5 = 3.2), alkene CH (generally 4 — 4.5 - 6) and ara- matic/heteroaromatic CH (Sy ~~ 9.5), und also between n-electron-rich (pyrrole, furan, thiophe- ne, d= 6- 7) and n-electron-deficient heteroaromatic compounds (pyridine, d= 7.5 - 9.5).12 2_RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR 24.2 Deuterium exchange Protons which are bonded to heteroatoms (X/ protons, X = O, N, S) can be identified in the 'H NMR spectrum by using deuterium exchange (treatment of the sample with a small amount of 1,0 or CD,OD). After the deuterium exchange: ROK + Uw Rox + HOO the XH proton signals in the 'H NMR spectrum disappear. Instead, the HDO signal appears al approximately 4, = 4.8. Those protons which can be identified by D;O exchange are indicated as such in Table 2.1, As a result of D;0 exchange, XH protons are often not detected in the 'H NMR spectrum if this is obtained using a deuteriated protic solvent (e.g. CDyOD). 24.3 8C Chemical shifts ‘The "C chemical shift ranges for organic compounds ** in Table 2.2 show that m containing functional groups can be identified by the characteristic shift values in th spectra For example, various carbonyl compounds have distinctive shifts. Ketonic carbonyl functions appear as singlets falling between & = 190 and 220, with cyclopentanone showing the largest shift, although aldehyde signals between fg = 185 and 205 averlap with the shift range of keto carbonyls, they appear in the coupled "C NMR spectrum as doublet CP? signals. Quinone car- banyls occurs between Bg ~ 180 and 190 while the cahoxy C atoms af eathoxylic avid sand their derivatives are generally found between = 160 and 180. However, the °C signals af phenoxy carbon atoms, carbonates, ureas (carbonic acid derivatives), oximes and other imines also lie at about 6¢ = 160 so that additional information such as the empirical formula may be helpful for structure elucidation. Other functional groups that arc casily differentiated are cyanide (8¢ = 110-120) from isocyanide (Sc = 135-150), thiocyanate (6c = 110-120) from isothiocyanate (¢ = 125-140), cyanate (Be 105-120) from isocyanate (Bc = 120-135) and aliphatic C atoms which are handed to different heterocstoms or substituents (Table 2.2). Thus ether-inethoxy generally appears between & = 35 aml 62, ester-iethoxy al Bg ~ $2; N-methyl generally lies belween 8g = 30 and 45 and S-methyl at about d¢ = 25. However, methyl signals at 8; = 20 may also arise from methyl groups attached to X or C=C double bonds, e.g. as in acetyl, CHj-CO- Tf an [7 atom in an alkane R-H is replaced by a substituent X, the °C chemical shifi 8 in the a position increases proportionally to the electronegativity of X (-/ effect). In the p-position, 8¢ generally also increases, whereas it decreases at the C atom 7 to the substituent (7-effect, sce Sec~ tion 2.3.1), More remote carbon atoms remain almost uninfluenced (48e-~ 0). NMR Ce wes ¢ FBG OOG. RX Siac bb: 0 -0 <0 90 >02.1. Functional groups 13 In contrast to 'H shifts, °C shifts cannot in general be used to distinguish between aromatic and heteroaromatic compounds on the one fiand and alkenes on the other (Table 2.2). Cyclopropane carhon atoms stand out, however, by showing particularly small shifts in both the "C and the 'H NMR spectra. By analogy with their proton resonances, the °C chemical shifis of m electron- deficient heteroaromatics (pyridine type) are larger than those of ek rings (pyrrole type). ciron-rich heteroaromatic Tabla 2.2.6 chemical shill anges for organic eampounds —& 20 160 100 50 ors) Cabeniunions — | Ketones orjugoto | | ery Tete corustg hha] anes | Catonpcacisandaane.| | | | — thioureas | i Ureas, Carborates | Oximes 1 | imines = | Iscyanses 14 Cries | Istioorentes i —_ Thecyerstee } 1] me yyy soja Carboaiides | — Heterosromatics sbledtoh defor! election | (op egung | | Aromatics (jaca {Cyolo } Alkynes | Hy0-Hologen 1 lk none 0 met | — —& 0 m0 100 0 crs) Al4 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR Substitucat cffects (substitucnt increments) tabulated in more detail in the literature '* demonstrate that °C chemical shifts of individual carbon nuclei in alkenes and aromatic as well as heteroaro- matic compounds can be predicted approximately by means of mesomeric effets (resonance cf fects). Thus, an election donor substituent D [D = OCHs, SCI, N(CHL)a] attached to a C=C dou- ble bond shields the f-C atom and the B-proton («M effect, smaller shift), whereas the e-position is deshielded (larger shift) as a result of substituent electronegativity (1 effect). = Be | bee | HD [aoa ni Donorin a shields in postion ‘Aoooptorin a doshiolds in postion The reversed polarity of the double bond is induced by a x electron-accepting substituent A (A = -O, C=N, NO,): the carbon and proton in the B-position are deshielded (-M effect, larger shifts). These substituents trave analogous effects on the C atoms of aromatic andl heteroaromatic rings. An electron donor D (see above) attached to the benzene ring deshieldls the (substituted) c=C stom (Feffect). In contrast, in the ortho and para positions (or comparable positions in heteraararneaic rings) it causes a shielding (4M effect, smaller 'H and "°C shifts), whereas the meta positions re- main almost unaffected. (+t substtuent (election donor D) bonded to the benzene rng: OHS T.20 . Oc 120.9 An electron-accepting substituent A (see above) induces the reverse deshielding in ortho and pura positions (-M effect, larger 'H and ''C shifis ), again with no significant effect on mera positions. (7-tesuttiuant (action acceptor) bonded tote benz S472 5 Bc ~ 128.5 2.1.4 *N Chemical shifts Frequently the "N chemical shifts “" (Table 2.3) of molecular fragments and functional groups containing nitrogen complement their 'H and '°C shifts, The ammonia scale ’ of ''N shifts used in21 Functional groups Table 2.3 shows very obvious parallels with the TMS scale of °C shifts. 16 1us, the 'SN shifts (Tab le 2.3) decrease in size in the sequence nitroso, nitro, imino, amino, following the corresponding behaviour of the "°C shifts of carbonyl, carboxy, alkenyl and alkyl carbon atoms (‘Table 2.2), Table 2.3, “N Chemical shift ranges for organanitrogen compounds = 900 200 700 00500400 300200100 OYA) onarese- ,—_ | Teiowes | | | | ten | | aides | mm ire | ‘20 compounds | Diez iazwniver a fe oe Pyne aes ones imieos nes mt yeas yroies \ || | | Cyanidee | | | Iecoanides | | | im Cuanidnes brn) ta fer Suforamiges Thins amises Annes Amines LL Lee ~~ 900 woo 700 600 500400 300 200 1000 (NH) The decrease in shifts found in °C NMR spectra in the sequence Bynenesammaies > Sanynr > Syaaner > Sapien also applies to the: amlogous N-containing functional groups, ring systems and partial structures, (Tables 2.2 snl 2.3) Simines, pyridine > Sites > Samines > Sasiidines16 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 2.2. Skeletal structure (atom connectivities) 2.21 HH Multiplicities The splitting (signal multiplicity) of 1H resonances oflen reveals the: spatial proximity of the pro- tons involved. Thus it is possible to identify structural units such as those which afew occur in organic molecules simply from the appearance of multiplet systems and by using (he (n +1) rule. The simplest cxample is the AY or AB two-spin system tor all substructures containing two pro- tons bwo, three or four bonds apart from cach other, according to geminal, vicinal ot w coupling. Figure 2.1 shows the three typical examples: (a) the AX system, with a buge shift difference: (ug v4) between the coupled protons H* and H” in relation to their coupling constant J,y; (b) the 4B system, with a smaller shift difference (v,_¥4) of the coupled nuclei (#' and H*) relative to their coupling constant Jin, and (c) the 4B system, with a very small shift difference [(vie-Vvi) < Jas] verging on the A> case, whereby the outer signals are very strongly suppressed by the roofing effect (AB effet). Figure 2.2 shows the 'H NMR partial spectra of a few more structural units which can casily be identified. Structure elucidation does not necessarily require the complete analysis of all multiplets in com- plicated spectra. If the coupling constants are known, the characteristic fine structure of the single multiplet almost always leads to identification of a molceular fragment and, m the case of alkenes and aromatic or heteroaromatic compounds, it may even Icad to the clucidation of the complete substitution pattern chomicaly non-equivalent gominal protons [eyabottanca.altones) yn \ HA ‘1200 "1800 (Co raameonts with vicinal aky rotons | in | HK Yeo b Ak | cs an trans-ethenyl groups AA, we I enetin Ha WA bay tn ‘409 (dvtare-aronatios wi erto= én) ‘ri rots (amg). oy c" HA ga H800 SS 600 Figure 2.1. AX (AQ) systems and typical molecular fragments2.2 _ Skeletal structure (atom connectivities) 7 Srosrtant Sosy Partial spectrom axe (singe asec! A001 i gies ax) ll f x onrcrt Ax x ae il cobetod ame iil vl (BR casnet yan) “ WN, Horry ae | at x I Figure 2.2, Easy to recognise A,X, systems and their typical molecular freaments18 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 2.2.2 CH Multiplicities The multiplicities of "C signals due to ‘Jey coupling (splitting occurs due to CH coupling across cone bond) indicates the bonding mode of the C atoms, whether quaternary (RC, singlet S), tertia- ry (R\CH, doublet D), secondary (CH), triplet T) or primary (RCH, quartet Q). i . | i 4 1 a ppm 42 409 380 MSHI BO I Figure 2.9. J-resulved two-dimensional "C NMR spect series of a-pinane (1) [In (CM,),CO. 25 °C, 50 MHz] (a) Stacked plot; (b) contour plot2.2. Shultslal structure (atom connectivities) 19 Coupled "°C NMR spectra which have been cnhanced by NUE are suitable for analysis of CH multiplicities (gated decoupling) *°. Where the sequence of signals in the spectra ts too dense, evaluation of spin multiplicities may be hampered by overlapping. In the past this has been avoi- ded by compression of the multiplet signals using off-resonance decoupling * of the protons, More roxlers techniques include the .J-modulated spin-echo technique (attached proton test, APT) "and Jeresolved twordimensional °C NMR spectroscopy |, which both use Jémodulation “Figure 2.3, shows a series of J-resolved "C NMR specira af a-pinene (1) as a contour plot and ag a stacked plot. The purpose of the experiment is apparent, °C shift and Joy coupling con- stants are shown in two frequency dimensions so that signal overlaps occur less often. ‘The J-modulated spin-echo '"" and the more frequently used DEPT experiment "** are pulse sequences, which transform the information of the C// signal multiplicity and of spin-spin coup- ling into phase: relationships (positive and negative amplitudes) of the °C signals in the proton- decoupled °C NMR spectra, The DEPT experiment benefits from a '7/-"C polarisation transfer Which increases the sensitivity by up to a factor of 4, For this re provides the quickest way of determining the '\C'H multiplicities. Figure 2.4 illustrates the application af the DEPT technique to the analysis of the CH multiplets of a-pinene (1). Routinely the result will be the subepectrum (b) of all CH carbon atoms in addition to a further subspectrum (c), in which, besides the CH carbon atoms, the CH carbon atoms also show positive amplitude, whereas the -arbon atoms appear as negative. Quatemary C’ atoms do not appear in the DEPT subspectra; accordingly, they may be identified as the signals which appear additionally in the '# broadband decoupled "\C NMR spectra (c.g. spectrum a in Fig. 2.4). cou, this techni | | 31513264 230 209 116.1 Wes Ppm Figure 2.4. CH mutipicites of a-pinene (1) (nexadeuterioacetone, 25 °C, 80 MH2]. (a) ‘H broadband decoupled C NMR spcetrum; (b} DEPT subspectrum of Ci; (¢) DLPT subspectrum ofall C atoms which are bonded to H (CH and GHs positive, CHe negative) (d) an expansion of a section of (e). Signals fram twe quaternary C atoms, three CH unite, wo CH, unite and three CH, units can be secn20 2 _ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR Figure 2.4 illustrates the usefulness of CH multiplicities for the purpose of structure elucidation The addition of all C, CH, CH, and CH, units leads to a part formula CH, 20+ ICH + 2CH; + ICHy = Cyt Cally + Calle + Call = Ciallie which contains all of the H atoms which are bonded to C, Hence the result is the hydrocarbon part of the molecule, e.g. that of a-pinene (1, Fig. 2.4). Ifthe CI/ balance given by the CH multiplicities differs from the number of H atoms in the mole cular formula, then the additional // atoms are bonded to heteroatoms. The "C NMR spectra in Fiy, 2.5 show, for example, for isopinocampheol (2), CjoFf)s0, a quaternary C atom (C), four C/T units (CH), two CH, units (CjH,) aud three CH, groups (CyH)). In the CIT balance, Coll, one His missing when compared with the molecular formuba, Cjoff 0; fo conclude, the compound contains one OH group finmuta of the a \ 701 ppm 482 GL A2t 391385 34,2 29 28 29 Figure 2.8. CH multiplicities of isopinocampheol (2), Cia /sO [(CD),CO. 2 °C, 50 MHz}. (a) 'H broadband de- coupled °C NMR spectrum; (b) DFPT CM subspactrim: (¢) DEPT sunspectrum of all C atoms whieh are bon- 69d ta H (CH and CH, positive, CH: negative)22 Skeletal stucture (atom connectivities) a4 2.23 HH Coupling constants Since spin-spin coupling *° through bonds occurs because of the interaction between the magnetic moments of the atomic nuclei and the bonding electrons, the coupling constants *" reflect the bonding environments of the coupled nucei. In 'H NMR spectroscopy geminal coupling through ‘two bonds (‘Jir,) and vicinal coupling through three bonds (*Jyqi) provide insight into the nature of these bonds Table 2.4, Typical HH coupling constants (Hz) of some units in alicycles, alkenes and alkynes ** z ee Yue ews Uae gemins protons prowone with w-mitionships ; | nt ies ~ =H -20 Saw | 7 f 7 oes eee Geminal HH coup ton Sigg, depends characteristically on the polarity and hybridisation of the C on the conpling path and also on the substituents and on the HCH bond angle. Thus “Jy, coupling cim be used to differentiate between a cyclohexane (12.5 Hz), a cyclopropane (~4.5 H2) or an alkene (2.5 2), and to show whether electronegative heteroatoms are bonded to methylene groups (Table 2.4), In cyclohexane and norbomane derivatives the w-shaped arrangement of the bonds between protons attached to alternate C atoms leads to distinctive “iq coupling (w-coup. lings, Table 2.4) Vicinal IH coupling constants, ‘Jy, ate especially useful in determining the relative configurati- on (see Section 2.3.1). However, they also reflect # number of other distinguishing characteristics, eg, the ting size for cycloalkenes (@ low value for small rings) and the a-position of electronega- five heleraalams in heterocycles which is reflected by remarkably small coupling constants Vz, (Table 2.5) The coupling constants of nrtho (gy — 7 He), meter (yy — 1.5 Hz) and para protons Jin $1 72) in benzene and naplubalene ring, systems are especially useful in structure elucidation (Table 2.5). With naphthalene: xnd other condensed (hetero-) aromatics, a knowledge of ‘zig zag’ coupling Clay ~ 0.8 F) is helpful in deducing substitution patterns,2 2_RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR ‘Table 2.5. Typical HH coupling constants (Hz) of aromatic and heteroaromatic compountis ?* | un ne Sat o7 or 09 | The HH coupling constants of pyridine (Table: 7.5) reflect the positions of the coupling protons relative to the nitrogen ring, There is a particularly clear difference here between the protons in the 2- and 3-positions (‘Jjj; = 5.5 Hz) and those in the 3- and 4-positions (‘Ji; = 7.6 Hz). Similarly, HH coupling constants in five-membered heteroaromatic rings such as thiophene, pyrrole and furan can be distinguished because of the characteristic effects of the electronegative heteroatoms ‘on their ‘Jj,z; couplings (Table 2.5); in particular the “Jy coupling of the protons in the 2- and 3 positions, allow the heteroatoms to be identified (the more electronegative the heteroatom, the smaller is the value of “Vj. In the case of alkenes and aromatic and heterosromatic compounds, analysis of a single multiplet will ofien clarify the complete substitution pattern, A few examples will illustrate the procedure. If, for example, four signals are found in regions appropriate for benzene ring protons (54 ~ 6-9, four protons on the basis of the height of the integrals), then the sample may be a disubstituted benzene (Fig. 2.6). The most effective approach is to analyse a multiplet with a clear fine structure2.2 _ Skeletal structure (atom connectivities) 23 and as many coupling constants as possible, e.g. consider the threefold doublet at dy ~ 7.5 (Fig 2.6 a); it shows two ortho couplings (8.0 and 7.0 Hz) and one meta coupling (2.5 Hz); hence tela- tive to the H/ atom with a shift value of dy = 7.5, there are two protons in ortho positions and one in a met position, to conclude, the molecule is an ortho-disubstituted benzene (o-nitrophenol, 3). Figure 2.6. 11/ NMR spectra of disubstituted benzetm rinys [CDCIs, 25°C, 200 MH7] (a) o-Nutrophenal (3): (B) _menitrobenzaldehyde (4); (¢) 4,4"-dimethoxybenzil (5)2 2_RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR ‘A meta disubstituted benzene (Tig, 2.6 b) shows only two ortho couplings ("Js ~ 7.5 Hz) for one signal (64 =7.8) whereas another signal (sy ~8.74) exhibits only two meta couplings (‘Jj ~ 3.0 snd 2.5 Hz). Tn both cases one observes either a triplet (Sy =7.8) or a doublet of doublets (3 = 8.74) depending on whether the couplings (Jyy or ‘Jjy) arc equal or different, ‘The AA'XX" systems (Section 1.5) ** which are normally easily recognisable from their symmetry identify para-disubstituted benzenes such as 4,4"-dimethoxybenzil (5) or 4 substituted pyridines. This method of focusing on a 'H multiplet of clear fine structure und revealing as tmany HH coup ling constants as possible affords the substitution pattern for an alkene or an aromatic or a hetero- aromatic compound quickly and conclusively. One further principle normally indicates the gemi- nal, vicinal and w relationships of the protons of a molecule, the so called HH connecttvities, i. that coupled nuclet have identical coupling constants. Accordingly, once the coupling constants of ‘a multiplet haye all been established, the appearance of one of these couplings in another multiplet identifies (and assigns) the coupling partner, ‘his procedure, which also leads to the solutions to problems 1-12, may be illustrated by means of two typical examples. 25° 8h 85 05 (25 \"" He a MW | he = : ai i 6 ppm 70 Sh (oro) x ” a Hy (mata) | | ne = 05 He ts) | . |) | | 1, pe WM l| I Kr » ii S —— a 7 7 pom ro Figure 2.7. ‘H NMR spectrum of 34-dimethoxybenzaldenyde (6) [aromatic shit range, CDCI, 25 °C, (a) 100 Miz, (b) 200 Miz In Fig. 2.7 the 'H signal with a typical aromatic proton shift of 3 =7.1 shows a doublet of doub lets with J-values of 8.5 Hz (ortho coupling, “Jqy)) and 2.5 Hz (meta coupling, “Ji. ‘The ring proton in question therefore has two protons as coupling partners, one in the ortho position (8.522 Skeletal structure (atom connectvit Hz) and another in the meta position (2.5 Hz), and moreover these are in such an arrangement as to make a sccond ortho coupling impossible. Thus the benzene ring is 1,2,4-trisubstituted (6). ‘The ring protons form an AMY system, and, in order to compare the change of frequency dispersion and ‘coofing’ effects with increasing magnetic field strength, this is shown first at 100 MIlz. and then alsa at 200 MHy. The: para coupling “Iyy- which is less frequently visible, is also resolved. From the splitting of the signal al é,= 7.1 (H") a 1,2, 3etrisubstitutea! benzene ring (7) might have bbeen considered. In this case, however, the ortho proton (H*) would fave shown a second arthe coupling to the third proton (H’). ‘The application of the principle that coupled nuclei will have the same coupling constant enables the ‘17 NMR spectrum to be assigned completely (Fig. 2.7). The ortho coupling, Jus = 8.5 Hz, is ropeated at dy = 6.93 and allows the assignment of //"; the meta coupling, ‘Jia = 2.5 TIz, which appears again at 4, = 7.28, gives the assignment of I. The four signals in the '‘H NMR spectrum of a pyridine derivative (Fig. 2.8) show first that it is a 2- or S-monosubstituted derivative: a 4-monosubstituted pyridine would display an 44“XX" sy stem, The signal with the smallest shift (6y~ 7.16) splits into a threefold doublet with coupling constants 8., 4.8 and U.7 Hz . The two “Ji couplings of 8.1 and 4.8 Hz uncquivocally belong to a f proton of the pyridine ring according to Table 2.5. Step by step assignment of all three couplings, (Fig. 2.8) leads to a pyridine ring 8 substituted in the 3-position. Again, signals are assigned fol- lowing the principle that coupled nuclei will have the same coupling constant; the coupling, con- stants identified fiom Table 2.5 for thee proton at y— 7.16 are then sought in the other multiplets 4-88 868 aie Ne AK AK A ati ~ ys tite a oe 8 uae a a" 8 a oa is a7 4 pon 8G igure 2.8, ‘HNMR spectrum of S-bromopyridine (8) [CDCIy . 25 °C. 90 MHz}26 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 2.24 CH Coupling constants One-bond CH coupling constants Jey (‘Jey are proportional to the s character of the hybrid bon- ding orbitals of the coupling carbon atom, (Table 2.6, from left to right) “°", according to Jon = 500s (ty where s=0.25, 0,33 and 0.5 for sp’-, sp’- and sp-hybridised C atoms, respectively. With the help of these facts, it is possible to distinguish between alkyl-C (/en-~ 125 Hz), alkenyl- and aryl-C (icy ~ 165 Nz) and alkynyl-C (Jez ~ 250 Tz), e.g. as in problem 15. It is also useful for structure elucidation that Jy inereases with the electronegativity of the hetero- atom or substituent bound to the coupled carbon atom (able 2.6, from top to bottom). Table 2.6, Stuctural features (carbon hybridisation, electionegatiyity, rng size) and typical one-bond CH cour pling constants Joy (Hz) so i 15 © Hybnaisaton Aa Clectronegativity = N” \ N 140 180 205 NSSH 200 sq my we \, 7 So “9 us i An An Ring size fm 1M apn 428 H | J 0 S 13 sa 0 From typical values for Jgy compling, Table 2.6 shows In the chemical shifi range for aliphatic compounds cyclopropane rings (ca 160 Hz reflect large $ character of bonding hybrid orbitals) oxirane (epoxide) rings (ca 175 Hz additionally reflect electronegativity of ring oxygen atom); cyclobutane rings (ca 135 Tz); O-alkyl groups (145-150 Hz); N-alkyl groups (140 Hz); acetal-C atoms (ca 170 Heat 8¢ ~ 100); terminal ethynyl gronps (ex 250 Hy)2.2 _Skeletal structure (atom connectivities) 7 In the chemical shift range for alkenes and aromatic and heteroaromatic compounds enol ether fragments (furan, pyrone, isoflavone, 195-200 Hz); 2-unsubstituted pyridine and pyrrole (ca 180 Iz); ‘2-unsubstituted imidazole and pyrimidine ( > 200 Hz). Geminal CH coupling “Jo becumes more: positive with increasing CCH bond angle and with decreasing electronegalivily of the substituent on the coupling C. This property enables a dist tion to be made inter alia between the substituents om the benzene ring or between hetercistanns in five-ring heteroaromatics (Table 2.7). From Table 2.7, those “Jcy couplings which may be espe- cially clearly distinguished and diagnostic are: #-C atoms in imines (e.g. C-3 in pyridine: 7 Iz); a C atoms in aldehydes (25 Hz); substituted (noneprofonated) C atoms of term | ethynyl groups (40-50 7). Table 2.7. Structural features and geminal (wwo-bond) CI coupling constants Vey (Hz) *®"" Bond angle 709.9 co r H 62 Eloctronegative substituents on tho coupling © 1 28-90 Vicinal CH couplings “Jey depend not only on the configuration of the coupling C and H (Table 2.8; see Section 2.3.2), but also on the nature and position of substituents: an electronegative sub- stituent raises the Joy; coupling constant on the coupled C and lowers it on the coupling path, e.g in alkenes and benzene rings (Table 2.8). An imino-N on the coupling path (e.g. from C-2 to 6-H in pyridine, Table 2.8) is distinguished by a particularly large “Jey coupling constant (12 Hz). In the °C NMR spectra of henzene derivatives, apart from the Jey, only the meta coupling (Vey, but not “Yor) is usually resolved, A henzenvidl CH, from whose perspective the meta positions are substituted, usually appears as a “Jey, doublet without additional splitting, e.g. in the case of 3,4- dimethoxy-p-methyl-P-nitrostyrene (9, Fig. 2.9) the carbon atom C-5 generates a doublet at 8: = 111.5 in contrast to C-2 at 8, = 113.5 which additionally splits into a triplet. The use of CH coup ling constants us criteria for assigning a resonance to a specific position is illustrated by this ex- ample.28 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR why” MeO md 123) Fey ke ban 360 ) Figure 29. "°C NMR spect of 3.4.dimetheny-p-methylp-nivostyrene (9) [CDCI 25 °C, 20 MH. (a, B) bveuadband decoupled, (a) complete specinum with CHy quarts at Bg = 11 and 56.0; (b, €) decoupled and coupled partial spectrum of benzenold and alkene carbon atoms, (¢) obtained by ‘gated! decoupling Assianments: © te tovn) Mlle JewtHe) Mil Yona) Caney so) ows ‘ 80 am cz 13560 uta os 3s m (6H RCH) co ys m Her. 400H) css tea cs tate Hath ca 9a aE HOH BCH) oe ‘ur? 75 cH wr Gwe 60 (Oo 0 tas The quotation marke indicate thatthe coupling constants ae vitual the seme for non-equivalent protons. C- shoul, for exam spit into adeuble (cso ac} of quartets Jot Ch since Both coupings have the eame value (7-5 He), « peoudoquinta ‘gut fe observed22. Skeletal structure (atom conncetivites) 29 Table 2.8 Siructural features at vival Uew-boel) CH coupling asmstarts Vey (He) Redatve cunliguration ane | TSH cH r9H Electronegative substituents on the coupling © Nite rs OAL clio L AL | sot NH 86 NH 78: Electronogative substituents on the coupling path 49) y° Hae oH Br Lone pair of electrons on imino-N on the coupling path HST N her Usually there is no splitting between two exchangeable XH protons (X = Q, N, 5) and C atoms: through two or three bonds ("Jen or “Jeu, unless solvation or an intramolecular Jf Inidge fix the XH proton in the molecule, Thus the C atoms ortho to the hydroxy group shew ‘cy coupling (o the hydrogen bonding O// proton in salicylaldchyde (10), whose values reflect the relative confi- gurations of the coupling partners. This method may be used, for example, to identify and assign the resonances in jrrablea 17. fl senanr I Nay, © OC 2.2.5 NH Coupling constants Compared with 'H and '°C, the magnetic moment of 'N is very small and has a negative value, The NH coupling constants are correspondingly stualler and their signs are usually the reverse of comparable 1/H and CH couplings **. Table 2.9 shows that the one-bond NH coupling, Jhns is proportional to the s-character of the hybrid bonding orbital on N so a distinction can be made between amina- and imino-NH. Formamides can be identified by large “Vy couplings between "Nand the formyl proton, The ‘Jy; and ‘Jy; couplings of pyrrole and pyridine are especially distinctive andl reflect the orientation of the non-bonding electron pair on nitrogen (pyrrole: per- pendicutar to the ring plane; pyridine: in the ring plane; Table 2.9), a fact wiich can be exploited in the identification of heterocyclic compounds (problems 30 and 31).30 2 RCCOGNITION OF STRUCTURAL FRAGMENTS 8Y NMR Tablo 2.9. Structural features and typical NH coupling constants (Hz) ” ca yee . Jun Now 270 NS Oe —C=N—H 1882 Hse { i He, 4 fh (SSeHts stm ‘Ws Has — 2.2.6 HH COSY (geminal, vicinal, w-relationships of protons) The HH COSY experiment **™” in proton magnetic resonance is a quick altemative to spin decoupling * in structure elucidation. 'COSY" is the acronym derived from carrelation speciras- copy) HH COSY correlates the 1H shifts of the coupling protons of a molecule. The proton shifis are plated on both frequency axes in the two-ditnensional experiment. The result is a diagram with square symmetry (Fig. 2.10). The projection of the one-dimensional 'H NMR spectrum ap- pears on the diagonal (diagonal signals). In addition there are correlation ot cross signals (off- diagonal signals) where the protons are coupled with one another. Thus the HH COSY diagram indicates HH connectivities, that is, geminal, vicinal and w-relationships of the H atoms of a mo lecule and the associated structural units, An IIH COSY diagram can be shown in perspective as a stacked plot (Fig. 2.10a). Interpretation of this neat, three-ditensional representation, where the signal intensity gives the third dimension, can prove difficult because of distortions in the perspective. The contour plot can be interpreted more easily. This shows the signal intensity at various cross-sections (contour plots, Fig. 2.10b). However the choice of the plane of the cross-section affects the information provided by an HH COSY diagram; if the plane of the cross-section is too high then the cross signals which are weak are lost, if it is too low, then weaker artefacts may be mistaken for cross signals. Every //// coupling interaction can be identified in the HA COSY contour plot by two diagonal signals and the two cross signals of the: coupling pariners, which form the four comers of a square, ‘The coupling partner (cross signal) ofa particular proton generates a signal on the vertical or hot vonlal line from the relevant ‘H signal. In Fig. 2.10b, for example, the protons at dy =7.90 and 7.16 are found as coupling partners on both the vertical and the horizontal lines from the proton 2 H of quinoline (11) at 4, = 8.76, Since 2-H (dj, = 8.76) and 3 H (4, = 7.16) of the pyridine ring in AL can be identified by the common coupling “Vij ~ 5.5 He (Table 2.5), the HH relationship which is likewise derived from the HH COSY diagram confirms the location of the pyridine protons in Ma, Proton 4-H of quinoline (8, ~ 7.0) shows an additional cross signal at 5, = 8.03 (Fig. 2.10). If it is known that this so-called zig-zag coupling is attributable to the benzene ring proton 8-112.2 Skeletal structure (atom connectivities) at (1b), then two further cross signals from y= 8.03 (at 54 = 7.55 and 7.35) locate the remaining (Ie) oO s a a 8 a e oe @ 8 » @ a s = ae b "875 809 oy 160-755 735 745 Figure 2.10. HiH/ COSY digaram of quinoline (11) [(CDs),CO, 95% viv, 25°C, 400 Miiz, 8 scans, 256 exper ents}, (a) Stacked plot; (b) contour plot92 2 _RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR | | i Wie a Zor Tse me 1H i dee Fu Ga et oom tte 2a an 221 20F Figure 2.11, Proton-Protun shill corations of axpinene (1) [purty 99 %, COCIy, % vv, 2b "C, BUY Mz, 8 scans, 256 experiments), (a) HH COSY; (b) HH TOCSY; (¢) selective one-dimensional fl? TOCSY, soft pulse itadiation at dy = 5.20 (signal nl shown), compared withthe ’H NMR spectrum on top; deviations of chemical Shits from those in other experiments (Fig. 2.14, 2.16) arise from solvent effects22 _ Skeletal structure (atom connectivities) 33 This example (Hig, 2.10) also shows the limitations of the HE COSY experiment: first, evaluation, without taking known shifts and possible couplings into account, is not always conclusive because the cross-sectional arca of the cross signals may not reveal which specific couplings are involved: sccond, overlapping signals (¢.g. 04 = 7.55 and 7.60 in Fig. 2.10) arc not separated by HH COSY if the relevant protons couple to one another. If there is sufficient resolution, however, the fine structure of the multiplets may be recognised hy the shapes of the diagonal and cross signals, ¢.. in Fi, 2.10, a 6y— 7.55 there is a tiple, therefore the resonance at éy= 7.60 is a doublet (sce the shape of the signal on the diagonal at §— 7.557 60 in Big. 2.10) In the case of (m+1}- and (n+1}-fold splittings of dy systems in one-dimensional 'H NMR spec tra the HH COSY plot gives, depending on the resolution, up to (m+1)(n+7)-fold splittings of the cross signals. If scvcral small coupling constants contribute to a multiplet, the intensity of the cross signals in the HH! COSY may be distributed into many multiplet signals so that even at a low-lying cross-section no cross signals appear in the contour diagram, Despite these limitations, structural fragments may almost always he derived by means of the JJJ COSY, so that with comp- lementary information fiom other NMR experiments it is possible to deduce the complete structu- re. Thus the HA COSY is expecially useful for solving the problems 12, 32, 33, 38, 46 and 52. Modifications of the basic HH COSY " include "COSY 45" with an initial 15° pulse to suppress diagonal signals for a better separation of cross signals near the diagonal or "COSY with delay" to emphasise conncetivitics which arc the result of smaller couplings. The "TOCSY" experiment (from total correlation spectroscopy) * detects cross signals of an individual proton with all other protons coupled to each other within a larger structural unit, e.g, a ring, the range depending on the adjustable time of isotropic magnetisation transfer. Thus, the endo-methylene proton of u-pi- nene at 6, = 1.16 displays only one cross signal with the exo-methylene proton (y= 2.34) due to geminal coupling in the HH COSY experiment (Fig. 2.1 1a), whereas additional eras sigels with 41> 1.94,2.07,2 17nd 221 appear in the HH TOCSY plot (Fig. 2.1 1b), thus revealing a conside- rably larger partial structure, One-dimensional HH TOCSY experiments involve irradiation with a selective ("soft") pulse adjustable to any non-overlapping proton signal of the molecule. Fig. 2.11 ¢, for example, detects all protons of the cyclohexene substructure including the methylene group in a one-dimensional HH TOCSY with selective irradiation of the alkene proton (y ~ 5.20). 2.2.7 CC INADEQUATE (CC bonds) Once all of the CC bonds in a molecule arc known, then its carbon skeleton is established. One way to identify the CC bonds would be to measure "*C-'"C coupling constants, since these a same for C atoms which are bonded to one anather: identical coupling constanls are known to identify the coupling partners (see Sevtivw 7.2.3), Unfortunately, this method is complicated by two factors. fst, 8C-"C couplings, especially those in the aliphatic region, are nearly all the same (lec = 35-40 Hr ', Fig, 2.12), provided that none of the coupling C atoms carries an electronega- live substituent, Second, the occurrence of ''C-'°C coupling requires the two nuclei to be directly bonded. However, given the low natural abundance of "C (1.1 % oF 107), the probability of a "e-"6 bond is only 10 *, Splitting a3 a result of “C-"C coupling therefore appears only as a weak feature in the spectrum (0.5 % intensity), usually in satellites which are concealed by noise at a distance of half the “C-'"C coupling constant on either side of the °C-'?C main signal (99 % intensity). ‘The one-dimensional variations of the INADEQUATE experiment "7 suppress the intense “C-"C main signal, so that both 4X and AB systems appear for all °C-!°C bonds in one spectrum. ‘The two-dimensional methods '*""*?!2? segregate these 4B systems on the basis of their indivi the34 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR dual double quantum frequencies (the sum of the °C shifts of A and B) as a second dimension. Using the simple cxample of I-butanol (12), Fig. 2.12a demonstrates the use of the two- dinensional INADEQUATE experiment for the purpose of structure elucidation. Por every bond the contour diagram gives an AB system parallel to the abscissa with double quantum fre- arbon bonds of 1-hatanol quency as ordinate. By following the arrows in Fig, 7 17a, the carbe can be derived immediately. The individual AB systems my also be plotted one-dimensionally (Fig. 2.12b); the "“C-"'C coupling constants often provide useful additional information. 32, ma. mo we 12 “Cy che “Cheon om 0 m3 82 co ko Figure 2.12, Two-dimensional (20-JNADEQUATE diagram of 4-butanol (12) [(CD,),C0, 95% viv, 25°C, 50 MHz, 128 scans, 128 expesiments) (a) Contour plot with the AB systems of ponded C atoms on the horizontal ‘axis; (b) plots of the three AB systems of the molecule obtained from (a); (e) contour plot of the symmetrised INADEQUATE experiment showing the AB or AX sysiems of honded C atoms in the HH COSY format (cross signais on the axes perpendicular to the diagonal) A Nariation on the INADEQUATE technique, sefereed tw as symmettised’ 2D INADEQUATE ®, provides a representation in the HH COSY format with its quadratic symmetry ofthe di gonal and cross signals. Hete the one-limensional 'H broadband decoupled ''C NMR spectrum is projected on lu the diagonal and the AB systems of all C-C bonds of the molecule are projected on (o individual orthogonals (Fig. 2.12¢). Every C-C bond then gives a square defined by diagonal siynals and off-diagonal AB patterns, and itis possible to evaluate as described for HH COSY ‘A disadvantage is the naturally low sensitivity of the INADEQUATE technique. However, if one has enough substance (more than § mg per C atom, samples fiom syntheses), then the sophisticas22. Skeletal structure (atom connectivities) 36: ted experiment is justified as the solutions to the problems 21,22, 35,36 and 37 illustrate, Fig. 2.13 is intended to demonstrate the potential of this technique for tracing out a bicyclic carbon skeleton ug the example of isopinocampheol (2). The evaluation of all CC-AB systems on the orthogo- nals leads to the eleven C-Chonds ack. If all the C-C bonds which have been found are combined, then the result is the bicyelie system (ich) and the three methyl substituents (i-k) of the molecule 2 and “\pm \A4 3 2 \ uo | | [| _ ~} 3 6 we 9a Mt as 85 En Figure 2.13. Symes tu-simensional INADEQUATE experiment wth 'sapnosampneo (2) (CD00, 250, ‘main 0.3 ml, 25°C. SOMIlz, 256 scans and exe]. (a) Stacked pot ofthe section between b= 20.9 ar 48: (b) ‘complete contour plot with cross signal pairs iabelled a-k for the 11 CC bonds of the molecule to faciitate the assignments sketched in formula 236 2 RECOGNITION OF STRUCTURAL FRAGMENTS 8Y NMR The point of attachment of the OH group within the molecule (at &¢ ~ 70.7) is revealed by the DEPT technique in Fig. 2.5. Figure 2.13 also shows the 48 effect on the "C signals of neighbou ring C atoms with a small shift difference (bond g with 8c = 47.4 and 48.2): the intense inner sig- uals appear very clearly, the weak outer signals of the AB system of these two C atoms are barely’ recognisuble excep! as dots. Additional cross signals without resolved doublet structure, e.g. bet- ween 8 — 48.7 aul 47.1, are the result of smaller 2Je¢ and “Jee couplings. 2.2.8 Two-dimensional carbon-proton shift correlation via one-bond CH coupling The CH COSY technique *"*"?* with carbon-13 detection and proton decoupling, and more sen- sitive and thus time saving methods with 'H detection and 'C decoupling denoted as inverse CH COSY, correlate “°C shifts in one dimension with the 'H shifts in the other via one-bond CH coup- ling Jew ‘The pulse sequence which is used 10 reeord CH COSY invalves the /H-2C pola which is the basis of the DEPT sequence and which increases the sensitivity by a faclor of up (o four. Consequently, a CH COSY experiment does not require any more sample than a 'H broad~ band decoupled "\C NMR spectrum. The result is a two-dimensional CH correlation, in which the "C shift is mapped on to the abscissa and the ’H shift is mapped on to the ordinate (or vice versa). The "C and 'H shifts of the '# and '°C nuclei which are bonded to one another are read as coord nates of the cross signal as shown in the CH CUSY stacked plot (Fig, 2.14b) and the associated contour plots of the a-pinene (Fig, 2.14a and ¢), To cvaluatc them, one need only read off the coordinates of the corrclation signals. In Pig. 2.14e, for example, the protons with shifts dy 1.16 (proton 4) and 2.34 (proton B of an AB system) are bonded to the C atom at 6c = 31.5. Formula 1 shows all of the C/J connectivities (C/T bonds) of aepinene which can be read from Tig. 2.14 HC HMC (heteronuclear multiple quantum coherence) and AC HSQC (heteronuclear single quantum coherence) are the acronyms of the pulse sequences used for inverse carbon-proton shitt correlations. ‘These sensitive inverse experiments detect onc-bond carbon-proton connectivities within some minutes instead of some hours as required for CH COSY as demonstrated by an IC HSUC experiment with o-pinene in Fig. 2.15 Carbon-proton shift correlations are particularly attractive for structure elucidation because they allow the shifts of two nuclei ('H and '"C) to be measured in a single experiment and within a feasible time scale, They determine all CH bonds (the CH connectivities) of the molecule, and hence provide an answer to the problem as to which HT nuclei are bonded to which "°C nuclei, The 'H multiplets, which so frequently overlap in the ‘AH domain, are almost always separated in the second dimension because of the larger frequency dispersion of the ""C shifts. ‘This proves to be particularly advantageous especially in the case of larger molecules, a feature illustrated by the identification of several natural products (problems 43-55). The resolution of overlapping AB systems as in the case of ring CH groups in steroids and in di- and triterpenes is especially helpful (problem 51). If there is sufficiently good resolution of the proton dimension in the spectrum, it may cyen be possible to recognise the fine structure of the ‘7 multiplets fiom the shape of the correlation signals, a feature which is useful for solving problems 32, 48 wud 51 Lion transfer Figure 2.14 (page 37), CH COSY diaaram of a-pinene [[CD:}C0. 10 % vv. 25 °C. 60 MHz for #0, 200 MHz for 'H 64 scans, 756 experiments} (a) Complete contour plat, () Stackes pot of te signals for the section outlined in (a) ftom Be = 20.9 to 47.2 (°C) ane A, = 0.85 Wo 2.34 ("HY (c) rontour plot of section (b), showing ene- dimensional °C and ‘H NM epecia for this section aligned paral! tothe abscissa and the ordinate22. Skeletal siucture (alc connectivities) a, Sep 18 6 2.08 a a 9 238 c pen 2D asm3 eh B2938 2 _RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR : >.) 09 ° > BO - ~ th et _ aa = ° F315 at + Ws ° 41? 5.20 Figure 2,15. HC HSQC experiment (contour plat) of u-pinene [(CDCls, 5 % viv, 25 °C, 125 MHz fur "*c, 900 MHz fur ‘H, 4 scans, 256 experiments]. This experiment gives the same information ag Fig. 2.14 within 8 minu- tes instead of two hours required for the CH COSY in Fig. 2.14 duc to higher sensitivity because of proton de- tection and stronger magnetic field. Deviations of proton shifls fam those in Fig 7.14 aulse frum the change of the solvent. 1he methylene protons collapsing in Fig. 2.14 at 5, = 2.19 (200 MHz) display in this experiment an AB system with By ~ 2.17 andl dy = 2.21 (500 MH?)22 _ Skeletal structure (atom connectivities) ED 2.2.9 Two-dimensional carbon-proton shift correlation via long-range CH coupling Two-dimensional CH correlations such as CH COSY or HC HMQC and HSQC provide the ‘Joy connectivities, and thereby apply only to those C atoms which are linked to H and not to non- protonated C atoms, Modifications of these techniques, also applicable to quatemary C atoms, are those which are adjusted to the smaller “Joy and “Jcy couplings (2-25 Hz, Tables 2.8 and 2.9) '**, Experiments that probe these couplings include the CH COLOC ** (correlation via long range couplings) with carbon-13 detection (Fig, 2.16) and HC: HMBC (heteronuclear multiple bond coherence) with the much more sensitive proton detection (Fig. 2.17)". 149 |» te} in ° 206 2 pm ur, 109 38 315313 264 230209 Figure 2.48. CH COLOC experiment of w-pinene I{CDsbCO, 10% viv, 25 *C. 50 Mllz for 0, 200 MHz for 1H, 288 scans and experiments, (a) stacked pict of te section between 8: = 20.9 and 47.2 (°C) and 3 = U.85 and 2.34 ('H};(b) contour plot of (a), One-dimensional °C and 'H NMIR spectra for this section are shown aligned With the abscissa and ordinal of he contour plot“ eatretation signals which are sready known from the CH ‘COSY study (Hig, 2.14) and have not been uppressed, are indicated by circles40 2 _RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR ‘These two-dimensional CH shift correlations indicate CAT relationships through two and more bonds (predominantly “Jew and “Jey connectivities) in addition to more or less suppressed ‘Ji relationships which arc in any casc established from the CH COSY contour diagram. Format and analysis of the C/Z COLOC or [IMBC plots correspond to those of a CH COSY or HSQC experi- nent, 2s is shown for vepinene (1) in Pigs. 2.14 - 2.17, When trying to establish the CH relationships of a carbon atom (exemplified by the quatemary © at 8¢~ 38.0 in Figs, 2.16 and 2.17), the chemical shifts of protons at a distance of two or thr bonds are found parallel to the proton axis (e.g. dy ~ 0.89 (0.84), 1.16 and 1.27 in Figs, 2.16 and 2.17). It is also possible to take a proton signal as the starting point and from the cross signals parallcl to the carbon axis to read off the shifts of the C atoms two or three bonds distant respecti- vely. Thus, for example, one deduces that the methyl protons at dy = 0.84 and 1.27 arc two and three bonds apart from the C atoms at c= 38.0, 40.9 and 47.2 as illustrated by the partial structu- ves 1a and Ub in Fig. 2.17. CI/ correlation signals due to methyl protons prove to he especially reliable, as do trans CH telationships over three bonds, e.g. between 44 ~ 1.16 nnd o ~ 38.0 ins Figs 2.16 andl 2.17, in conteast to the snissing civ relationship between Ay — 2.34 and Bo — 38.0 (partial structures 1d and 1y in Fig. 2.17) due to the smaller coupling constant to which the expe- riment was not adjusted. : ' i. bts i 2% ‘ pom 161 pom Figure 2.16 (6), CH COLOG waperiment of a-pinene [(CD:):CO, 10 % viv, 25 °C, 80 MHz for °C, 200 MHz tor 'v, 256 ecans and experiments], complete contour plot (caption on previous page)22 _ Skeletal structure (atom connectivities) 4 ese es be bbe - 58 @ M64 4a ° e 2 buss 2H" 207m 167 LPT 066 ppm 221 2iF Figure 2.17. HC HMBC experiment of a-pinene [(CDCh, § % viv, 25 *C, 125 MHz for “C, 500 Mle for Hy, 16 Scans, 258 experiments, contour plat], This experiment gives tre same information ag Fig, 2.16 within 24 min instead ot 8 hrs required for the CH COLOG in Fig. 2.18 duc to higher sensitivity because of proton detection and stronger magnetic field. Deviations of proton shills turn those In Fig 2 18 anise from the change of the solvent. ihe methylene protons collapsing in Fig, 2.16 at 3, =2.19 (200 MHz) display in this experiment an AB syste will 4 ~ 2.17 ani dy = 2.21 (500 M7)42 2_RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR 2.3. Relative configuration and conformation 2.31 HH Coupling constants Vicinal coupling constants jz, indicate very clearly the relative configuration of the coupling, protons. Their contribution depends, according to the Karplus-Conruy equation >, Vin = acon? = 0.28 (upto. = 90". a= 10; above = 90% a~15) (2) ‘on the dihedral angle ¢, enclosed by the CZ bonds as shown in Fig, 2.18, which sketches the Kar- plus-Consoy curves for dihedral angles from 0 to 180°. Experimental values correspond to those ven by (he curve shows; deviations are up to 3 Hz; electronegative substituents on the coupling path, for example, reduce the magnitude of yey Hz paz 18 18 16 | 16 4 14 a n2 12 10. bo 8 be 64 L6 4 4 2 2 o ° 0 30 60 90 120 150 180° e jgure 2.18. Vicinal HH coupling constants “ho, a function ofthe ditedral angle of the GH bonus con ‘corned (Karplus-Conroy relationetip} For the stable conformers 13a-¢ of a substituted ethane the vicinal HH coupling constants J, = 3 Hz for syn-protons and J, ~ 15 Hs for anti-protons can be derived from Fig, 2.18. If there is rotation around the C-C single bond, the coupling protons pass through the syn configuration twice and the anti configuration once. 0" 9 = 180° H syn (gaucha) anti (rans) ~ syn (ganecha) 13a 13b 132.3 Relative configuration and conformation 2 Therefore, from “Ivo = CA + I)/3 = 213 = 7 (3) an average coupling constant of about 7 Hz is obtained. This coupling constant characterises alkyl groups with unimpeded rotation (cf. Figs |.2 and 2.19). Ethyl dibromodihydrocinnamate (14), for example, can form the three staggered conformers Maee by rotation around the CC single bond a. to the phenyl ring. 9 6 = 180" @ =" 4 4 4 Hse 2020. ‘| a 8 vy 00st He ps ar OTe |Orr sm goer) an ans, veo) an gous} 14a 14b 14c ‘The “HNMR spectrum (Fig. 2.19) displays an AB system for the protons adjacent te this hon; he: coupling constant \J4p = 12 Hz. From this can be deduced first that the dihedral angle o between the CH bonds is about 180°, second that conformer 14b with minimised steric repulsion between the substituents predominates and third that there is restricted rotation around this CC bond. Ree ppm 73 $3 48 43 13 0 Figure 2.49. /1 NMR spectrum of ethiy! dibromodiiydocinnanate (44) (CDCl, 25°C, 8 Mr, CW recording]“4 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR The relative configuration of the protons which is deduced from the coupling constant ‘Jj con firms the conformation of this part of the structure of this molecule. On the other hand, the ‘Vim coupling constant of the ethyl group attached to oxygen (7 Hz, Fig. 2.19) reflects equal popula- tions of all stable conformers around the CC bond of this ethyl group. The Vj couplings shown in Table 2.10 verify the Karplus-Conroy equation 2 (Fig, 2.18) for rigid systems. Hence in cyclopropane the relationship ‘Uji; > “Vrrrnnonsy holds, because cis-cyclo- propane protons enclose a dihedral angle of about 0°, in contrast to an angle of ca 145° between trans protons, as shown by Dreiding models. Vicinal protons in cyclobutane, cyclopentane, nor- borane and norbornene behave in an analogous way with larger cis, endo endo and exo exo coup. lings, respectively (able 2.10), Substituent effects (electronegativity, configuration) influence these coupling constants in four-, five- and seven-membered ring systems, sometimes reversing the cis-trans relationship ’*, so that other NMR methods of structure elucidation, e.g. NOE difference spectra (see Section 2.3.5), are needed to provide conclusive results. However, the coupling, constants of vicinal protons in eyclo- hexane and its heterocyclic analogues (pyranoses, piperidines) and also in alkenes (Table 2.10) are particularly informative. Neighbouring diavia! protons of cyclohexane can be clearly identified by their large coupling con- stants (Jgg 11-13 Hz, Table 2.10) which contrast with those of protons in diequatorial or axial equatorial configurations (‘Ure ¥ “Use * 2-4 Hz). Similar relationships hold for pyranosides as oxy- gen hetero analogues of cyclohexane, wherein the electronegative O atoms reduce the magnitude of the coupling constants (J, = 9 1Iz, ge = 4 Iz, Table 2.10). These relationships are used for elucidation of the configuration of substituted eyclohexanes (problem 37), cyclohexenes (pr lems 12 and 35), teipenes (problems 47, 48, 50 and 51), flvans (problem 10) and glycosides (ptoblem 45, Table 2.10). In these cases also, the relative configuration of the protons which is deduced from (he ‘Jj; coupling, constant reveals the conformation of the six-membered rings. ‘Thus the coupling constant 9 Hz of the protons in positions | and 2 of the methyl-B-D-gluco- pyranoside 15 determines not only the diaxial configuration of the coupling protons but also the °C, conformation of the pyranose ring. If the sterically more crowded 'C, conformation (exclusi- vely with axial substituents) were present then a diequatorial coupling (4 Hz) of protons J-H and 2-H would be observed. If the conformers were inverting (50: 50 population of the “C; and 'Cy conformers), then the coupling constant would be the average (6.5 Hs) : 0 HO. on 15 "GX ie ey We The couplings of vicinal protons in 1,2-isubstituted alkenes lie in the range 6-12 Hz for cis pro- tons (dihedral angle 0°) and 12-17 Hz for srans protons (dihedral angle 180°), thus also following the Karplus-Comoy equation. Typical examples are the alkene proton 4B systems of coumarin (la, cis) and rrans-cinnamic acid (16b), and of the cis-trans isomers 17a and b of ethyl isopente nyl ether, in addition to those in problems 3, 4, 8, 11, 13 and 382.3. Relative configuration and conformation 46 ax ox SHz(c1s) yg = 155 He (rans) 16a 16b 2a Hote CHICHabe Het) HA of et CHICHaie Supa = 0 He (us) 17a 17b 2yag = 13 He ans) Table 2.10. “J coupling constants (Hz) and relative configuration **. The coupling path is shown in bold ysleprapanos yotanes ‘cup H cont Naat p ‘Norbamanes e y 99 NN 80 4 er0-090- onde onde Cyctieranes nibequatarial Pranoses Bo Rw eo ‘keno » zt HH o-92 ende-oxo Sees YER 10 i 4 slewvatuial iat ore, WoL oor Hiei =46 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 2.3.2 CH Coupling constants Geminal CH coupling constants “Jey, reflect the configuration of electronegative substituents in molecules with a defined geometry such as pyranose and alkenes '*, If an electronegative substitu- ent ig attached cis with respect to the coupling proton, then the coupling constant “Joy has a higher negative value; if it is located frans to the coupling proton, then “cy, is positive and has a lower value: this is illustrated by Band a D glucopyranose (18a and b) and by bromoethene (19). wot wi , 18a 18b 19 Vicinal CH coupling constants Voy vesemble vicinal HH coupling constants in the way hat they depend on the cosine” of the dihedral angle o between the CC bond to the coupled C atom and the CH bond to the coupled proton “* (cf. Fig, 2.16), as illustrated by the Newman projections of the conformers 20a-c of a propane fragment. 6 = 100° x H Jon 2 Ja B5 Si 2 20a 20b 20¢ Ti follows from this (hat where there is equal population of all conformers in an alkyl group (free rotation about the CC single bond) then an averaged coupling constant “Jey = (Qin + Sans) /3 of between 4 and 5 Hz can be predicted, and that vicinal CH coupling constants “Uc have values about nwo thirds of those of vicinal protons, “Jynr '. Like “Vy couplings, “Jey couplings give conclusive information concerning the relative configa- ration of C and J/ as coupled nuclei in cycluhexane: aul pyranose rings snd in alkenes (Table 2.11), Substituted eyctohexanes have Voy, = 2-4 Hz for cis and 8-9 Hz for trans configurations of the coupling partners; electronegative OH groups on the coupling path reduce the magnitude of ‘Jou in pyranoses (Table 2.11), When deducing the configurations of multi-substituted alkenes, e.g. in solving problem 19, the ‘Joy; couplings of the alkenes in Table 2,1 | are useful. eso > Veuies) holds throughout. Tlectronegative substituents on the coupling carbon atom increase the J-value, whilst reducing it on the coupling path, Moreover, ‘Jey reflects changes in the honding stale (carbon hybridisation) and also steric hindrance (impeding coplanarity), as ther examples in Table 2.11 show.23 Relative configuration and conformation 47 Table 2.11. “lq, coupling constants (Hz) and relative configuration "The: coupling path is shown in bald yclohcxane derivatives and pyranosas mh ote faa wane ena Alwanes carbon nyortsation cis 15H Cs 1a a tron aon not sari “city | Eiostanogatvo subatuents ee G41 HCHp-Br on tho coupling C ABH CHy ros aon agit “Br anime eouning pam | Steric interactions 4 4 i 104 4 a5H Hee Yo fom aw iw Oe now oa | 2.3.3 NH Coupling constants The relationship between “Jay and the dihedral angle of the coupling nuclei, of the type that applies to vicinal couplings of ‘77 and "C, very rarely permits specific configurational assignments because the values (Uy < 5 Hz) are too small 7, Tu contrast, geminal couplings “Jyy; distinguish the relative configurations af aldimines very clearly. Thus, anti-furan-2-aldoxime (21a) shows a considerably larger “Jy coupling than does the syn isomer 21b; evidently in imines the non- bonding electron puir cis to the CH bond of the coupled proton has the effect of producing a high negative contribution to the geminal NH coupling. Wea Buh (re) How ©. ay ate 2ta 21b"°8 2 RCCOGNITION OF STRUCTURAL FRAGMENTS BY NMR 23.4 C Chemical shifts ‘A carbon atom in an alkyl group is shielded by a substituent in the y-position, that is, it experien- ces a smaller '°C chemical shift or a negative substituent effect *. This originates from a sterically induced polarisation of the CH bond: the yan der Waals radii of the substituent and of the hydro- gen atom on the ¥-C overlap; as a result, the ¢-bonding clectrons are moved from H towards the y- C atom; the higher electron density on this C atom will cause shielding. As the Newman projecti- ons 22a-e show, a distinction can be made between the stronger 7-syn and hie weaker punt effect If there is free rotation, then the effects are averaged according (@ the usual express (2% You/3, and one observes a negative y-substituent effect of -2.5 to -3.5 ppm **, which is typical for alley! groups. Ton 22a 22 In rigid molecules, strong 7 effects on the “U shift (up to 10 ppm) allow the different configura tional isomers to be distinguished unequivocally, as cis- and trans-3- and -4-methyleyelohexanol (able 2.12) illustrate perfectly: if the OM group is positioned axial, then its van der Waals repul- sion of a coaxial H atom shiclds the attached C atom in the y-position, /.3-Diaxial relationships between substituents and /Y atoms in cyclohexane, norbornane and pyranosides shicld the affected C atoms, generating smaller "°C shifts than for isomers with equatorial substituents (Table 2.12) The "C chemical shift thus reveals the relative configuration of substituents in molecules with a definite conformation, e.g, the axial position of the OH group in trans 3-methyleyclohexanol, cis- 4-methyleyclohexanol, [-D-arabinopyranose and a D-xylopyranose (able 2.12, p. 50). It turns out, in addition, that these compounds also take on the conformations shown in able 2.12 (ara~ binopyranose, 'C. ; the others, “C,); if they occurred as the other conformers, then the OH groups on C-1 in these molecules would be equatorial with the result that larger shifts for C-1, C-3 and C-5 would be recorded. A ring inversion (50: 50 population of both conformers) would an average "C shift. Compared with 'H chemical shifts, '°C shifts are more sensitive to steric effects, as a comparison of the ’H and the "C NMR spectra (cis- and trans-4-tert-butyleyclohcxanol 23) in Fig, 2.20 shows. The polarisation through space of the y-CI7 bond by the axial OI/ group in the: eis isomer 23b shiclds C-I hy -5.6 and €-3 hy 4.8 ppm (y-efTect). In contrast the ‘H shifts reflect the con- siderably smaller anisotvopic: elect (see Section 2.5.1) of cyclohexane bonds: equatorial substitu- ents (in this case Hand OF) display larger shifts than axiai substituents; the equatorial 1-H in the is-igomer 231 (4,=3.92) has a larger shift than the axial 1-H in the trans-compound 23a 40); the difference is significantly smaller ( 0.52 ppm) than the y-effect on the °C shitts (ca -5 ppm). Both spectra additionally demonstrate the value of NMK spectroscopy for quantitative analysis of mixtures by measuring integral levels or signal intensities, respectively. Finally, D;O ‘exchange eliminates the OH protons and their couplings trom the ‘HW NMR spectra (Fig. 2.20d).23. Relative configuration and conformation 0 400 34 ppm trans as 39 307 us a W% ans 279 ass 28 a mo aa 23a Ha 23b on sor 1% oe n% 20%) Loot lal PF. mr ppm ido 366 38629. 783 trans 64 188 were 78 ans cis eve Figure 2.20. NMR spectra of trans-and cis-4-ter-butylcyclohexanol (28a and 2b) [(CDsjC0. 25 °C, 400 MHz for 'H, 100 Miz for "C], (a) 'H decoupled "c NMR spectrum (NOE uppressed, comparable signal intensities) (b) ‘HNMR spectrum: (¢) section of (b) (dy = 3-4) wilh integrals, (W) partial spectrum (c) following M0 ‘change. The integrals (¢) and the "°C signal intensities (a) give the trans : cis isomer ratio 71 : 29, Proton 4-H (a 9.40) nthe trans isomer 23a forms a triplet (10.8 Hz, two anf protons in 2,2-poxitons) of quate (3 H7, {io sym protons In 2,2-positions and the OH proton as additonal Coupling partner; folowing U,O exchange triplet (10.8 He) of tplets (4.3 He) appears, because the coupling to O!/is missing. Inthe cis isomer 296 proton 4-H forms a sexlel (3.0 He, four synetinal potons in 2.7-postions and OH) whieh appears as a quintet following 1,0 exchange because the coupling to UH is then lost90 2 _RCCOGNITION OF STRUGTURAL FRAGMENTS BY NMR ‘Table 2.12. "C chemical shifts () an relative configurations of cycloalkanes, pyranoses and alkenes (appl- cation of effects)“ ‘The shifts whict are jstinted in boldface reflect y-effects on C atoms in the corresponding ‘somer pare HS 204 ns 84 mz sa 2s mee ON }-Methyicyolohexanol DIHg- RE trans- Ge BSH A AEM atic yenrol Aaa ms c on as ne ani maine: trans TNH tetramer! ay = ea ar ou on 36 D-Arbinoyarose ay, 0) a pu (Cy entoration) : on © a oC Bie HD OH Hom Xt82_o D-Xyinpyrannse - Oat OM ey conormaton) o va at 4 ssa ny | ean OA baa 128 2atenyneronmane si mB oxo. 2.Norbomarel nds 14 ma 05 He Ba a “hs noo The y-cffect on the "C shift also causes (Ie: difference between (E)- and (Z)-configurations of the alkyl groups in alkenes. Here: the C-ur atom shift responds most clearly 10 the double bond confi gorational change; these atoms in cis-alkyl groups occupy y-positions with respect to each other; they are eclipsed, enclosing a dihedral angle of 0°, which leads to an especially strong van der ‘Waals interaction and a correspondingly strong shielding of the "°C nucleus, For this reason, the relationship [Sra > dr] holds for the a-C’ atoms of alkencs, as shown in Table 2.12 for (E)- and (2) 2 hexene. The °C shitts of the doubly bonded carbon atoms behave similarly, although the effect is considerably smaller.23. Relative configuration and conformation st a,f-Unsaturated carbonyl compounds show smaller °C shifts than comparable saturated com pounds “*, provided that their carbonyl and CC double bonds are coplanar. If steric hindrance prevents coplanarity, conjugation is reduced and so larger '°C shifts are observed, In o,8-un- saturated carbonyl compounds such as benzophenones and benzoic acid derivatives the twist angle 8 between the curbonyl double bond and the remaining a-system can be read off and hence the conformation derived from the “C shift”, as several benzoic acid esters (24) illustrate. ete anzen —e p * R bog O° e\ H 18800 He0y 4 oly 170449 CHiCHse 1713.57 ClCHya 1734 0 2.3.5 NOE Difference spectra Changes in signal intensities caused by spin decoupling (double resonance) are referred to as the clear Overhauser effect (NOF) *#°. In proton decoupling of "C NMR spectra, the NOT. increa~ ses the: intensity of the signals generated! by the C atoms which are bonded to hydrogen hy up to 300 %; almost all techniques for measuring "°C NMR spectra exploit this gain in sensitivity 7° 12 in recording 'H NMR spectra certain proton resonances are decoupled (homonuclear spin decoup- ling), then the changes in intensity due to the NOE are considerably smaller (much less than 50 %). Tor the assignment of configuration itis uscful that, during perturbing the equilibrium population of a particular proton by irradiating it for long enough, other protons in the vicinity may be affec- ted although not necessarily coupled with this proton, As a result of molecular motion and the dipolar relaxation processes associated with it, the popnlations of energy levels of the protons change ™*, their signal intensities change accordingly (NOB). For exemple, if the signal intensity ‘of one proton increases during imadiation of another, then these protons must be positioned close to one smother in the molecule, irrespective of the number of bonds which separate them, NOE difference spectroscopy has proved to be a useful method for studying the spatial proximity of protons in a molecule ””. In this experiment the ‘JJ NMR spectrum is recorded during the: irra diation of a particular proton (measurement 1), an additional measurement with an irradiation frequency which lies far away (the ‘off-resonance’ experiment) yal iso conditions, is then the basis for a comparison (reference measurement 2). The difference between the two measurements provides (hee NOE difference spectrum, in which only those signals are shown whose: intensities are increwsed (positive signal) or decreased (negative signal) by NOE. Figure 2.21 illustrates NOE difference spectroscopy with a-pinene (1): irradiation of the methyl protons at dy = 1.27 (experiment c) gives a significant NOE on the proton at dy ~ 2.34; if for comparison, the methyl protons with ¢ = 0.89 ate perturbed (experiment b), then no NOE is observed at 5, = 2.34. From this the proximity of the methylene-H atom at é, ~ 2.34 and the me- thyl group at 1.27 in a-pinene is detected. In addition, both experiments contirm the assignment of the methyl protons to the signals at 2, ~ 0.89 and /.27. A negative NOE, as on the protons at 5y = 1.16 in experiment e, is the result of coupling, c.g. in the ease of the geminal relationship with the affected proton at diy ~ 2.44, Further applications of NOE difference spectroscopy are provided in problems 29, 33, 34, 36, 44, 48, and 50-54,v2 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR 1 goth Ati i b Paes wu — A ” cone g g 1S 5 ’ : ror 0 (OR 8 ay Ba pom 234 219 206 1.93 1a 127116 Figure 2.21. Hr NOE difference spectra (b. ¢) and HH NOESY diagram (d) of u-pinene (1) with 'M NMR spec ‘tum (a) for comparison [(CD3):CO, 10% viv, 28 °C, ZOU MHz, section Irom 2 ~ 0.85 to 2.37] Vertical arrows in (b) and (c) indicate the iradiation frequencies; in the HHY NOESY plot (d), crass-signals linked by a dulled line show the NOF detected in (¢)23. Relative confiquration and conformation 63 2.3.6 HH NOESY and ROESY The HH COSY sequence for ascertaining the HH connectivities also changes the populations of the energy levels, leading to NOFs. Thus, the HH’ COSY experiment has been modified to pulse sequences such as HH NOESY '** which allows a two-dimensional detection of NOEs. The re- sult of such measurements is shown in the HH NOESY plot with square symmetry (Fig, 2.214) which is evaluated in the same way as HH COSY. Fig. 2.21d shows by the cross signals at éy 2.34 and 1,27 that the appropriate protons in a-pinene (1) are close to one another; the experiment also illustrates that the HAT COSY cross signals (duc to through-bond coupling) are not completely suppressed. Therefore, before evaluating @ two-dimensional NOE experiment, it is essential to know the /Z// connectivities from the III COSY plot. The NOESY sequence sensitively reacts towards the change of the sign of NOEs depending on molecular motion. The ROBSY experiment *" involving an isotropic mixture of magnetisations by spin locking detects NOEs without these limitations. Thus, the proximity of the methyl protons in e-pinene ((d, ~ 0.84 in CDCI.) to the exo-methylene proton ( ~ 2.//) is additionally indica- ted and assigned in the ROESY experiment (Fig. 2.22). Problems 46 and 55 exemplify further applications. 234 207 19% 167 tie 6.84 tn ot Figure 2.22, HH ROESY diagram of a-pinene (1) with 'H NMR spectrum [CDCh, 10% wv, 25 °C, 500 MHz, section from 4, = 0.84 to 2.34], Deviations ot chemical shtts trom those in other experiments (Figs, 2.14, 2.16) arise from solvent effects: the methylene protons collapsing in Fig. 221 al d ~ 2.19 (200 MHZ) dlsplay i this ‘experiment an AB systom with 3, = 2.1/ and dy ~ 2.27 (800 MHz)of 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR A comparison of the methods of proton-proton NOE detection has shown that two-dimensional NOE detection such as NOESY and ROESY are better suited to the investigation of the stereo chemistry of biopolymers whereas for small- to medium sized molecules (up to 30 C atoms) NOE difference spectroscopy is less time consuming, more selective and thus more conclusive 2.4 Absolute configuration 2.4.1 Diastereotopism Where hoth // atoms of a methylene group cannot be brought into a chemically identical position by rotation or by any other movement of symmetry, they are said to be diastereotopic **. The precise mening of diastereotapisi is est illustrated by means of an example, that of methylene protons HH and H® of glycerol (25). Where there is free votation about the CC bonds, the terminal CH.OH groups rotate through three stable conformations. They are best shown as Newman pro= jections (25a-c) and the chemical environments of the CH,O protons, HP and H® are examined ‘with particular reference to geminal and synelinal neighbours. 25a 25b on He 0 we HoH cron cron ction He: OH, HP OH, GHZOH: 5, (OF HY; GHLOH HP: dy OH. HR: HE, OH 8 HR: HE, OH; CHZOH HE: & HALO: HE, OH: 8 HH OH; OH ,CHOH: 3, It can be seen that the six possible near-neighbour relationships are all different, TC rotalion were frozen, then three different shifts would be measured for H" and Hin each of the conformers a, b and ¢ (4), 6; and 6; for H“, &,, 6; and d; for H"). If there is free rotation at room temperature and if ay %p and ye are the populations of conformers a, b and ¢, then according to the equations 4, Be Halt patie + Yat and — p= Bali bet + Heli (4) different average shifts 5, and dy are recorded which remain differentiated when all three confor- ‘mations occur with equal population (24 = x» = x2 = 1/3). Chemical equivalence of such protons would be purely coincidental. Figure 2.23 shows the diastereotopism of the methylene protons (CH*H°OD) of glycerol (25) in D0 solution (OF exchanged to OD); it has a value of dp ~ 04 = 0.09. The spectrum displays an (AB),C system for the symmetric constitution, (CH“#OD),CH°OD, of the molecule with geminal coupling “J4x~ 1.6 Hz and the vicinal coupling constants °Jj4:— 6.4 and “Uw: ~ 4.3 Hz. The un- equal “J couplings provide evidence against the unhindered rotation about the CC bonds of gly cerol and indicate instead that conformer a or ¢ predominates with a smaller interaction of the substituents compared to b.24 Absolute configuration 55 pase —on 25 Ho-aue CuaHe: 2, = 350 & - 359 % = 379 oH Figure 2.23. 'H NMR spectrum of alyceral [25, D,O, 10 %, 25 °C, 400 MHz Diastereotopism indicates prochtrality, as exemplified by glycerol (25, Fig, 2.23). Other examples of this include diethylacetals, in which the OCH; protons are diastereotopic on account of the prochiral acetal-C atoms, thus forming 4B systems of quartets because of coupling with the me thyl protons, ‘The Newman projections 2Sa-e draw attention to the fact that the central C atom, as seen from the terminal CH,OD groups, appears asymmetric. It follows from this that diastereotopism is also 2 way of probing neighbouring asymmetric C atoms. Thus the methyl groups of the isopropyl resi- dues in D- or L-valine (26) are diastereotopic and so show different ‘H and ‘°C shifts, although these cannot be individually assigned to the two groups. In chiral alcohols of the type 27 the dia- stercotopism of the isopropyl C nucleus increases with the size of the alkyl residues (methyl < isopropyl < tert butyl)”. Boy Hy 7H 098 oJ LH Hy 4 CHICHs)e ‘ey 198 105 cean Ifa molecule contains several asymmetric C atoms, then the diastereomers show diastereotopic shifts. Cliomasteral (282) and sitosterol (28b) for example, are two steroids that differ only in the absulute configuration at one carbon atom, C-24 ™. Differing shifts of "C nuclei close to this asymmetric C atom in 28a and b identify the two diastereomers including the absolute configura. tion of C-24 in both, The absolute configurations of carboxylic acids in pyrrolizidine ester alka- loids are also reflected in diastereotopic ’H and °C shifts *!, which is used in solving problem 54,56 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR a ee Ae? 288 cionasieri (745) 28b siosteroi zany 2.4.2 Chiral shift reagents (ee determination) The presence of asymmetric C atoms in a molecule may, of course, be indicated by diasterentopic shifts and absolute configurations may, as already shown, be determined empirically by compari- son of diastereotopic shifts *"'. However, enantiomers are not differentiated in the NMR spec- trum. The spectrum gives no indication as to whether a chiral compound exists in a racemic form or as a pure enantiomer. Nevertheless, it is possible to convert a racemic sample with chiral reagents into diastereomers or sirnply fo dissolve it in an enantiomerically pure solvent R or S; following this process, solvation diastereoiuers arise: from the ravemate (RP + SP) of the sample P, e.g. R-RP and R:SP , in which the enantiomers are recugnisable because of their different shifts. Corupounds with groups which influence the chemical shift because of their anisotrupy effect (see Sectioms 2.5.1 aud 2.5.2) are suitable for use as chiral solvents, e.g. I-phenylethylamine and 2,2,-triflworo-1-phenylethanol ” A reliable method of checking the enantiomeric purity by means of NMR uses curopium(III) or prasendymium(I) chelates of type 29 as chiral shift reagents **. With a racemic sample, these form diastereomeric eurupiwin(II]) or praseodyrnivia(I) chelates, in which the shifts of the two enantiomers are different, Different signals for & and S will be observed only for thase nuclei in immediate proximity to a group capable of coordination (OH, NH;, C=O). The separation of the signals increases with increasing concentration of the shift reagent; unfortunately, line broadening of signals due to the paramagnetic ion increases likewise with an increase in concentration, which limits the amount of shift reagent which may be used. Figure 2.24 shows the determination of an enantiomeric excess (ge) following the equation 5 Rs eo = 2S s 00¢%) o for |-phenylethanol (30) by ‘H and ''C NMR, using tris[3-(heptafluoropropylhydroxymethylene)- D camphoratopraseodymium(II1) (29d) as a chiral shift reagent ToS cits Hs R u 29a ca 29b pr24 Absolute configuration or mon m3 BE ana 1 Figure 2.24. Detention of the enantionvaic excess of t-phenyletranol[90, 0.1 mmol in 0.3 mi CDCb, 25 °C} by addition of the chiral praseodymium chelate 29b (0.1 mmol). (a, b) 'H NMR spectra (4U0 MHz), (a) without ‘and (b) with the shift reagent 28b. (c, d) °C NMR spectra (100 MHz). (c) without and (d) withthe shift reagent 29b. In the "C NMR spectrum (d) only the C-x atome of enantiomers 30R and 30S are resolved. Ihe 'H and ¢ signals of the phenyl residues are not shifted: these are not shown for reasons of space. The evaluation of the integrals gives 73 %% Rand 27 % S, ie. an enantiomeric excess (ee) of 46 %90 2_RECOGNITION Or STRUCTURAL FRAGMENTS BY NMR 2.5 Intra- and intermolecular interactions 2.5.1 Anisotropic effects The cheinical shift of a nucleus depends in part om its spatial position in relation to a bond ar a bonding system. The knowledge of such anisotropic effects ** is useful in structure elucidation, An example of the anisotropic effect would be the fact that aria? nuclei in cyclohexane almost always show smaller ‘H shifts than equarortal nuclei on the same C atom (illustrated in the soluti- ons to problems 37, 47, 48, 50 and 51), The y-effect also contributes to the corresponding beha- viour of ‘°C nuclei (see Section 2.3.4), Me pe re Multiple bonds are revealed clearly by anisotropic effects. Textbook examples include alkynes, shielded slong, the C=C tiple bond, and alkenes snd carbonyl compounds, where the muclei are deshielded in the plane of the C=C and C=O double bonds, respectively °' One criterion for di- stinguishing methyl groups attached to the double bond of pulegone (31), for example, is the car- bonyl anisotropic effect. } 31 2.5.2 Ring current of aromatic compounds Benzene shows a considerably larger 'H shift (64 = 7.28) than alkenes (cyclohexene, dy ~ 5.59) or cyclically conjugated polyenes such as cyclooctatetraene (4, ~ 5.69). This is generally explained by the deshiclding of the benzene protons by a ring current of x-electrons °° which is induced when an aromatic compound is subjected to a magnetic field. The ring current itself produces its ‘own magnetic field, opposing the external field within and above the ring, but aligned with it outside **. As a result, nuclei inside or above an aromatic ring display « smaller shift whereas nuclei outside the ring on a level with it show a larger shift. The ring current has a stronger effect ‘on the protons attached to or in the ring tht on the ring C atoms themselves, so that particularly TL shifts prove a useful means of detecting ring currents and as aromaticity criteria for investiga- ting annulenes. \ 4 Bo exteraimaet ring curent model for benzene fing caret att25_Intre- and intermolecular interactions 59 1,4-Decamethylencbenzene (32) illustrates the ring current of benzene by a shielding of the me- thylene protons (6, = 0.6) which lie above the aromatic ring plane in the molecule, A clear repre sentation of the ring current effect is given by [18Jannulene (33) at low temperature and the viny- ogous porphyrin 38 with « diaza{26Jannulene periseter * the inner protons are strongly shiel- ded (4) = -2.88 and -11.64, respectively); the outer protons are strangly deshielded (6, = 9.75 and 13.67, respectively), The typical shift of the inner NH! protons (6, = ~2 w —3) indicates that porphyrin 34 and its expanded analogues such as 38 occur as dixza{18Jannulene tautomers. Pro- blem 39 draws attention to this point. 2.5.3 Intra and intermolecular hydrogen bonding Hydrogen bonding can be recognised in ‘H NMR spectra by the large shitts associated with it; these large shifts are caused by the electronegativity of the heteroatoms bridged by the hydrogen atom, The OH protons of enol tautomers of 1,3-diketones are extreme examples. They form an intramolecular H bond and appear between dy ~ 12.5 (hexafluoroacetylacetone enol, Tig. 2.25) and 15.5 (acetylacctone enol) Intermolecular H bonding can be recognised in the 'H NMR spectrum by the fact that the shifts due to the protons concemed depend very strongly on the concentration, as the simple case of methanol (36) demonstrates (Fig.2.25a); solvation with tetrachloromethane as a solvent breaks down the hydrogen bridging increasingly with dilution of the solution; the OH shift decreases in proportion to this. In contrast, the shift of the ‘21 signal of an intramolecular bridging proton ve- mains almost unaffected if the solution is diluted as illustrated in the example of hexafluoroacety!- acetone (37), which is 100 % enolised (Fig. 2.25b). Intermolecular H7 bonding involves an exchange of hydrogen between two heteroatoms in two different molecules. The H atom does not remain in the same molecule but is exchanged. If its exchange frequency is greater than is given by the Heisenberg uncertainty principle, Mourn © tdyg (IB ~ 22a ®60 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR then its coupling J,y to a vicinal proton #7" is not resolved. Hence CH,, protons do not generally show splitting by vicinal SH, OH or NH protons at room temperature. ‘The same holds for Ui couplings with such protons. If the hydrogen bonding 1s hindered by solvatation or is intramole- cular, then coupling is resolved, as the cxample of salicylaldchydce (10) has already shown (sco Section 2.2.4; for applications, sce problems 17 and 25). He-0, 4 ii HN AA ets 36 oan ff 37 HW 4 eee) 7 =e % i j Jo - | \ ii 7 7 ; 00 opm "E ° Figure 2.25, 'H NMR spectra of metnanol (36, a) and hexatiuoroacetylacetona (37, b), both in the pure state (above) and dted in tetrachloromethane solution (6%. below) [25 °C. 90 MHz, CW recording] 2.5.4 Protonation effects If a sample contains groups that can take up or lose a proton, I" (Nib, COO), then ane rust expect the pH and the concentration to affect the chemical shift when the experiment is carried out in an acidic or alkaline medium to facilitate dissolution. The pH may affect the eheraical shift of ‘more distant, nonpolar groups, as shown by the amino acid aluine (38) in neutral (betaine form 38a) or alkaline solution (anion 38b). The dependence of shift on pH follows the path of titration curves; it is possible to read aff the pK value af the equilibrium from the point of inflection **.2.6 Molecular dynamics (ftuxionality) 6 182 Hy 128 38a Hse—CH—CO; = H:c—cH—co; 38D SNe aoa Nita pase pa 2 "SC shifis respond to pH changes with even greater sensitivity; this is demonstrated by the values nee (39I) ancl ifs cation (39a), mG) a oes 39a The effect of pH is rarely of use for pK measurement; it is more often of use in identifying the site of protonation/deprotonation when several basic or acidic sites are present, Knowing the incre- mental substitutent effects Z“* of amino and ammonium groups on benzene ring shifts in aniline and in the anilinium ion (40), one can decide which of the nitrogen atoms is protonated in proca ine hydrochloride (problem 24) 2.6 Molecular dynamics (fluxionality) 2.6.1 Temperature-dependent NMR spectra Figure 2.26 shows the ‘H NMR spectrum of N,N dimethylacetamide (41) and its dependence on temperature. At 55°C and below two resonances appear for the two N-methyl groups. Above 55°C the signals become increasingly broad until they merge to form one broad signal at 80°C. This temperature is referred to as the coalescence temperature, T-. Above T. the signal, which now be- fongs to both N-methyl groups, becomes increasingly sharp. The temperature-dependent position and profile of the N-methyl signal result from amide canoni- cal formulae of 41 shown in Fig. 2.26: the CN bond is a partial double bond; this hinders rotation of the N,N-dimethylamino group. One methyl group is now cis (8 = 3.0) and the other is trans (5 = 2.9) to the carboxamide oxygen. At low temperatures (55°C), the N-methyl protons slowly ex- change positions in the molecule (slow rotation, slow exchange). If energy is increased hy heating, (10 above 90 °C), then the N.N-dimethylamino group rotates so that the N-methyl protons «x= change their position with a high frequency (free rotation, rapid exchange), and one single, sharp Nemethyl signal of double intensity appears with the average shift (6p + 8,)/2 = 2.95o2 2 RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR Figure 2.26. ‘HNMR spacira of N,Nutimethytecetaricte (64) al the lenperatunes indicated [(CD;})80, 75 % vy, 80 MHz} The dimethylamino group rotation follows a firsorder rate law; the exchanging methyl protons show no coupling and their singlet signals are of the same intensity. Under these conditions, equa- tion 7” *“"! affords the rate constant k, at the coalescence point 7, ke = alve- wa) = eaw/p = 222 Av 0 where Avis the full width at half-maximum of the signal at the coalesence point 1; it corresponds to the difference in chemical shift (vy~v4) observed during slow exchange. In the case of dime- thylacetamide (41) the difference in the chemical shifi is 0.7 ppm (Fig. 2.26), ie. & Hz (at 80 MH?). From equation 7 it can then he calculated that the N-methyl groups at the coalescence point (80°C or 353 K) rotate: with an exchange frequency of k, — 2.22 x 8 = 17.8 Hz, According to the Fyring equation 8, the exchange frequency £, decreases exponentially with the free molar activati- on energy 4G 8) where R is the gas constant, & is the Boltzmann constant and h is the Planck constant. Equations 7 and & illustrate the value of temperature-dependent NMR for the investigation of molecular dyna- mics: following substitution of the fundamental constants, they give equation 9 for the free molar activation energy AG for first-order exchange processes: AG = 19.4 Te [ 10.32 + log (T2/ ki)1x 10-3 kd! mol fa)2.6 Molecular dynamics (fuxionaity) 63 ence the activation energy barricr to dimethylamino group rotation in dimethylacetamide (41) is calculated from equation 9 with k, = 17.8 sat the coalescence point 353 K (Fig, 2.26): Ass, = 78.9 kJ/mol oF 18.7 keal /mMol ‘Temperature: dependent (dynamic) NMR studies are suited to the study of processes with rate constants between 10"! and 10° s"' *, Some applications are shown in Table 2.13 and in problems 13and 14. Table 2.13. Selected applications of dynamic proton resonance **” Tek) Gy, | GH (kJ) mol) ‘enaney 7 Na “o _ ‘ Auxionaty) ‘COLHs Ss COREHy 2.6.2 '3C Spin-lattice relaxation times The spin-lattice relaxation time 7; i the time constant with which an assembly of a particular nuclear spin in a sample hecones rmagnetised parallel to the magnetic field us it is introduced into i. The sample: agnetisation Mp is regenerated after every excitation with this time constant. For organic molecules the 7; values of even differently bonded protons in solution are of the same order of magnitude (0.1 - 10 s). '"C nuclei behave in a way which shows greater differentiation between nuclei and generally take more time: in molecules of varying size and in different chemi- cal environments the spin-lattice relaxation times Iie between a few milliseconds (macromolecu Jes) and several minutes (quaternary C atoms in small molecules), With 'H broadband decoupling only one T; value is recorded for each C atom (rather than n T, values as for all n components of a complex 'H multiplet), and these "C spin-lattice relaxation times are useful parameters for pro- ‘bing molecular mobility in solution ‘The: technixjue for measurement which is most easily interpreted is the inversion-recovery method, in which the distribution of the nuclear spins among the energy levels is inverted by means of a suitable 180° radiofrequency pulse “*"’, A negative signal is observed at first, which becomes increasingly positive with time (and hence also with increasing spin-lattice relaxation) and which6 2_ RFCOGNITION OF STRUCTURAL FRAGMENTS BY NMR finally approaches the equilibrium intensity asyruptotically. Figures 2.27 and 2.28 show asympto: tical increases in the signal amplitude due to “C spin-lattice relaxation up to the cquilibrium value using two instructive examples. A simple analysis makes use of the: 'zcim intensity interval’, to, without consideration of standard deviations: after this time interval tp, the spin-lattive relaxation is precisely far enough advanced for the signal amplitude to pass through zero. Equation 10 then gives 7; for cach individual C atom. Ty = mlm2 ~ 1.45% (10) ‘Thus, in the series of 7; measurements of 2-octanol (42, Fig. 2.27) for the methyl group al the hydrophohic end of the molecule, the signal intensity passes through zero at ty = 3.8 s. From this, using equation 10, a spin-lattice relaxation time of 7; = 5.5 s can be calculated, A complete rela- xation of this methyl C ator requires about five times longer (more than 30 s) than is shown in the last experiment of the series (Fi. 2.27), 7; itself is the time constant for an exponential increase, in other words, after 7) the difference between tle: observed signal intensity and its final value is still Ve of the final amplitude. ro me fo 1a Ri ae RA DOS BS AM Tr ns 49 a9 30 22 22 39 22 AT, 96 08 78.60 ad ad 38 78 6 Figure 2.27. Sequence of measurements Ip dalermine tle: °C spin-alice relaxation times of 2-octanol (42) [(CD,),CO, 75% viv, 25 °C, 20 MHz, inversion-recovary sequanca, stacked plot}. The times at which the signals pass Iheough 7er0, %, have ben usta lo calculate, by equation 10, the T; values shown above for the "°C nuclei ‘ot 2 octanol2.6 Molecular dynamics (fluxionalty) 95 If smaller NOE cnhancements arc recorded for ccrtam ‘°C nucle, then other mechanisms (¢.g. spin-rotation) contribute to their spin-lattice relaxation ‘The main contribution to the spin-lattice relaxation of °C nuclei which are connected to hydrogen ig provided by the dipole-dipole interaction (DD mechanism, dipolar relaxation). For such °C nuclei a nuclear Overhauser enhancement of almost 2 will be observed during 'H broadband de- coupling according to: no = y/ 2ye = 1.988 un where yy and 7 are the gyromagnetic constants of "Hand "°C. ipolar relaxation of °C nuclei originates from the protons (larger magnetic moment) in the same or in neighhouring molecules, which move with molecular motion (translation, vibration, rotati- on), This motion generates Muctuating local magnetic fields which affect the observed nucleus. If the frequency of # local magnetic field matches the Tarot fiequency of the °C nuckens being observed (resonance condition), then this nucleus can undergo transition from the excited state (0 the ground state (relaxation) or the reverse (excitation). From this, it follows that the spin-lattice relaxation is linked to the mobility of the molecule or molecular fragment. If the average time taken between two reorientations of the molecule or fragment is defined as the effective correlati- on time +, and if m #/ atoms are connected to the observed C, then the dipolar relaxation time Tray is given by the correlation function: Top! = constant x nt (12) Accordingly. the relaxation time of a C atom will increase the fewer hydrogen atoms it bonds to and the faster the motion of the molecule or molecular fragment in which it is located. From this, it can be deduced that the spin-lattice relaxation time of '°C nuclei provides information concerning. four molecular characteristics: Molecular size: smaller molecules move: more quickly than larger ones; as a result, C atoms in sirall molecules relax more slowly than those in lirge molecules. The C atoms in the more mobile cyclohexanes (7; = 19-20 s) take longer than those in the more sluggish cyclodecane (7; = 4-5 s) = The number of bonded H atoms: if all parts within a molecule move at the same rate (the same +, for all C atoms), the relaxation times 1; deerease from CH via CH, to CH; in the ratio given by TH(CH) 2 Ty (CH) £1, (CH) = 62322 (13) Since methyl groups also rotate freely in otherwise rigid molecules, they follow the ratio shown in equation 13 only in the case of considerable steric hindrance °, In contrast, the 7; values of "C nuclei of CH and CH groups follow the ratio 2 : | even in large, rigid molecules. ‘Typical examp- les are steroids such as cholesteryl chloride (43), in which the CH, groups of the ring relax at approximately double the rate (0.20.3 s) of CH carbon atoms (0.5 s). Contrary to the prediction made by equation 13, freely rotating methyl groups require considerably longer (1.5 3) for spin- lattice relaxation.06 2_ RECOGNITION OF STRUCTURAL FRAGMENTS 8Y NMR ‘= Segmental mobility: if one examines the 7; series of 2-octanol (42, Fig. 2.27) calculated accor- ding to equation 10, it becomes apparent that the mobility parameters m7; increase steadily from C-2 0 C28, ASa result of hydrogen bonding, dhe: molecule close the: OM groups is almost rigid (nT, between 3.5 aul 4.4.) With increasing distance fron the anchoring effect of the OF group the mobility increases; the spin-lattice relaxation time becomes correspondingly longer. The nT; values of the (wo methyl groups also reflect the proximity to (7.8 s) and distance from (16.5 s) the hydrogen bond as a braking’ d & vag wor we tO 9 ppm c 26 3 4 55 35 cath 55 s Figure 2.28. Sequsnce of measurements to determine the spin-lattice relaxation times of the "°C nuclsi of the pyndine ning in L-nicotine (46) [(CD,),CO, 75 % viv, 25 °C, inversion-recovery sequence, 20 MI lz). The times at which signals pass through zero liave been used Iu calaulale, by eruation 10, me T; values for ie pyriaine C atoms in L-nieoting ® Anisotropy of molecular motion: monosubstituled benrene rings, e.g, phenyl benzoate (44), show a very typical characteristic: in the pura position to the substituents the CH nuclei relax con- siderably more rapidly than in the: urtho awl meta positions. ‘The reson for this is the anisotropy27 Summary 6 of the molecular motion: the benzene rings rotate more easily around an axis which passes through the substituents and the para position, becausc this requires them to push aside the least number of neighbouring molecules. This rotation, which affects only the o- and m-CH units, is too rapid for an effective spin-lattice relaxation of the o- and m-C atoms. More efficient with respect to relaxa~ tion are the frequencies of molecular rotations perpendicular to the preferred axis, and these affect the CH bond in p-positinn, Ifthe phenyl rotation is impeded by hulky substituents, e.g. in 2,2'6,6- tetramethylbiphenyl (48), then the 7, values of the CF atoms can be even less easily distinguished! in the mera and para positions (3.0 and 2.7, respectively). Particularly large spinlatice relaxation times obtained for non-protonated carbon nuclei of 45 and 46 arise from less efficient, non-dipolar relaxation mechanisms. . 6 EG. aD 44 45 46 Figure 2,28 shows the anisotropy of the rotation of the pyridine ring in nicotine (46). The main axis passes through C-3 and C-6; C-6 relaxes correspondingly more rapidly (3.5 s) than the three other C/T atoms (5.5 s) of the pyridine ring in nicotine, as can be sccn from the times at which the appropriate signals pass through zero, 2.7 Summary Table 2.14 summarizes the steps by which molecular structures can be determined using the NMR methods discussed thus far to determine the skeleton stmcture, relative configuration and confor- mation of a specific compound In the case of completely unknown compounds, the molecular formula is a useful source of ad tional information; it can be determined using small amounts of substance (a few micrograms) by high resolution mass spectrometric determination of the accurate molecular mass. It provides information concerning the double-bond equivalents (the ‘degree of unsaturation’ ~ the: nusuher of multiple bonds and rings). For the commonest heternatoms in organic molecules (nitrogen, oxy- gen, sulphur, halogen), the number of double-hond equivalents can be derived from the molecular formula by assuming thal oxygen and sulphur may be omitted and require no replacement atom, Inalagen tay be replaced by hydrogen and nitrogen may be replaced by CH, The resulting empiri- cal formula C,H, is then compared with the empirical formula of an alkane with m C atoms, C,Hoyi23 the number of double-bond equivalents is equal to half the hydrogen deficit, (n+ 2-x) 12 From CyH;NO (problem 4), for example, the empirical formula yf 13 derived and compared with the alkane formula Cy/z9; a hydrogen deficit of ten and thus of five double-bond equivalents is deduced, If the NMR spectra have too few signals in the shift range appropriate for multiple bonds, then the double-bond equivalents indicate rings (see, for example, a-pinene, Fig. 2.4)C 2_ RECOGNITION OF STRUCTURAL FRAGMENTS BY NMR |f the amount of the sample is sufficient, then the carbon skeleton is best traced out from the two- dimensional INADEQUATE experiment, If the absolute configuration of particular C atoms is needed, the empirical applications of diastereotopism and chiral shift reagents are useful (Section 2.4), Anisotropic and ring current effects supply information about conformation and aromaticity (Section 2.5), and plI effects can indicate the site of protonation (problem 24), Temperature- dependent NMR spectra and °C spin-lattice relaxation times (Section 2.6) provide insight into molecular dynamics (problems 13 and 14), Table 2.14. Suggested tactics for solving structures using NMR chemical enaisis, —s molecular formula o zi *Problem 46 141 Problem 46, continued (¢) HH TOCSY plot with 'NMR spectrum and selective HH TOCSY with soft pulse irradiation at diy = 5.70 (top); (d) HH ROESY plot with 'H NMR spectrum, same section and same scales as shown in (¢). 46d | i Wi Lah he of sow " o ; to | ho ie ” ® 4 | t : | | oe wen a | | of . «| id a 3 | o 8 | o atte af & | 4 A EEE EEEeE Ene eee eee]192 9 PROBLEMS Problem 47 ‘A compound with the elemental composition C, 00 Boo & 3 qe 66-8 0 0 2534 cia 0 o0 i | 654 0 i ws: Yoo > pon tat boa 388 u88 10 | 49d198 3 PROBLEMS Problem 50 ‘A compound with the elemental composition Cyyl/zsO¢ determined by mass spectrometry, was isolated fiom the Tight petroleum extract of the leaves of Senecio darwinii (Compositae, Hooker and Arnoll) ®, a plant which grows in Tiewa del Fuego (Chile). What structure can be derived from the set of NMR experiments 50? Conditions: CDChs, 25 mg per 0.3 ml, 25°C, 400 MHz. ('#2), 100 MHz.("C), (a) HH COSY plo; 50a