NMR

Course,

Lecture

14.29 April 14, 1999

Lecture

The

Chemical

Shift

Chapters (pages) in 'Modern NMR Techniques for Chernistry Research' by A. E. Derorne : .Notbing
~

unfortunately
papers related to the topics in this lecture : by P. w. NMR Atkins : Ch. 20.1 An Integrated Approach' by R. J. Abraham and P.

Other useful books or scientific .'Physical .'Proton Chemistry' and Carbon-13

Spectroscopy.

Loftus : Ch. 2 ~ .'Nuclear Magnetic Resonance Spectroscopy. A physicochemical View' by R. K. Harris : Ch. 1.8- 1.10 and Ch. 8.1 -8.16 .'N MR Spectroscopy' by H. Gtinther : Ch. 4.1 .Many books on chemistry, spectroscopy and physical chemistry in general cover the basics of NMR including chemical shifts and their origin

Page 111

NMR Lecture 4

14.55 April 12. 1999

With a slight oversimplification one can say that virtually everything that produces a magnetic field will influence the chemical shiit (the resonance frequency) of a given nucleus. The most important influence on the resonance frequency of a nucleus : -A nucleus is shielded by the electrons surrounding it -The electrons create a small magnetic field opposed to Bo. the external field coming from the magnet. due to the circular motion of the electrons around the nucleus induced by BO -Since the new field is opposed to BO the nucleus will 'feel' a magnetic field slightly smaller than BO Bett = BO -Bshield Bett refers to the 'felt' magnetic field. the one which determines the Larmor frequency. Vo -The above formula can also be written as Beft = BO -BO * 0"

Where 0" is the shielding constant From this we can see that the Bshield is dependendent on BO. the external field -The higher the electron densityaround our nucleus. the higher is Bshield or 0" and the more shielded our nucleus will be -For a hydrogen atom where the distribution of electrons is spherical (in atoms. does not happen in molecules) since there are only s-electrons the 0" is purely due to diamagnetic effects and therefore O"=O"d -In C W NMR the external field Bo must be increased to compensate for Bshield. Therefore. nuclei experiencing large Bshield are said to resonate upfields. In Fr NMR this corresponds to small ~values. Normally the right side of the spectrum -Accordingly. the left part of our spectrum is called the downfield region. BO is small in C W NMR and the ~values are high in Fr NMR In molecules and in atoms with p-ele(:trons (unsyrnmetric and non-spherically distributed), the total 0" of a nucleus is also affe(:ted by circulation of ele(:trons within the entire molecule

'--J

These 'molecular' electron movements cause a reduction of 0" for our nucleus and this is called a paramagnetic shift, O"p Paramagnetic effects (O"p-term) causes large downfield shifts (to higher vor ppm, to the left in our spectrum) If we knew the nature of electronic motion in a molecule (induced by BO) we could calculate allO" and subsequently all chemical shifts This we cannot do (except for H2 and LiR) and approximations are made to give a model which can be used to predict chemical shifts 0" = O"I~Cal + O"I~Cal+ 0", Here d docal and ~ocal are the local contributions to 0" for a particular nucleus and 0"' can be solvent effects, anisotropic effects, ring current effe(:ts etc. For protons the d docal and 0"' terms are most important since paramagnetic effects can not arise from its own valency electrons (no p-orbitals in H). The neighbouring atoms may however contribute to the ~ocal term of a proton

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NMR Lectln"e 4

14.55 April 12, 1999

The lack of p-orbitals on a hydrogen atom explains w hy virtually all protons resonate within -10 ppm while other nuclei resonate within hundreds or thousands of ppm (C -250 ppm, N 900 ppm, F 800 ppm, Co 18000 ppm etc )

How do all these theories affect a real spectrum? Practical considerations .Reducing electron density around a nucleus obviously reduces~ocal .This is very important for 1H resonances .In 'normal' molecules the H is however not bonded directly to a highly electronegative atom (F, Cl, O etc). Instead the proton is bonded to a carbon which in tum is bonded to the highly electronegative atom. The electronegativity of the substituentaffects the partia! charge on the carbon which then influences the shielding of the proton
",-..,

The O-values (in ppm) for the serles of compounds (CH3-X) below show that a more electronegative substituent (higher electron-withdrawing power) reduces the O'~cal for the protons of the methyl group and accordingly we seehigher chemical shifts, the signal moves
downfield

Si(CH3)4 0.00

CH3CH3 0.88

CH31 2.16

CH3Cl 3.05

CH30H 3.38

CH3F 4.26

-Reverse effects, with respect to (lH) versus electronegativity, CH3-CH2-X where X={F, Cl, Br, I}.

can be observed for

o(CH3-CH2-F) < o(CH3-CH2-Cl) < o(CH3-CH2-Br) < o(CH3-CH2-I) Other, geometrical, factors are responsible for this There is also an additive effect when several subtituents are present : -Higher
(""' , J

number of electronegative substituents on the <x-carbon add to the total deshielding of

thelH I CHCl3 I CH2Cl2 I CH3Cl I I 7.27 I 5.30 I 3.05 I The effect of the electronegativity of X decreases for protons further away. Protons bonded to the J3-carbon (3 bonds from X) or to the y-carbon (4 bonds) are less affected then protons bonded to the a-carbon (2 bonds)

"

-CH2-Br -CH2-CH2-Br -CH2-CH2-CH2-Br 3.30 1.69 1.25 -Linear relationships have been found and NMR chemical shifts have been used to measure electronegativities of a substituentX : EX = 0.684 (CH2-CH3) + 1.78 -Caution must be exercised since other effects, like the 'reverseeffect' for ethyl halides above, may appear, but within a small group of similar molecules the formula can be used

Page2115

NMR Lecture 4

14.55 April 12, 1999

Proton chemical shifts for substituted CH2XY and CHXYZ following formula and table : ~H = 0.23 + 1: contributions

groups can be predicted from the

Accuracy is approx. :i:O.3 ppm for CH2 groups, less accurate for the CH group
~

Ex. 1: (CH2CI2) Ex.2:

= 0.23+2.53+2.53

= 5.29 ppm = 1.37 ppm

Exp. = 5.30 Exp. = 1.25

(CH3CH2CH2CH2Br)

= 0.23+0.47+0.67

-The hybridization shift

state of the carbon atom to which a proton is bonded affects the chemical

H-v-.

H I ,..
H

H I
'"', ""

H /
H

'\
H

H-C=C-H

/c=c,

-Sp3 -Sp2

: mix ofl

s and 3 p orbitals ~ little s-character of the molecular orbital

: mix of 1 s and 2 p orbitals ~ more s-character of the molecular orbital

r-"

-sp : mix of 1 s and 1 p orbitals ~ much s-character of the molecular orbital -The more s-character the closer the electrons are to the C nucleus, and the further away from the H-nucleus. This results in a deshielding of the proton -Expected chemical shifts are therefore : o(ethyne) > o(ethene) > O(ethane)

THIS IS NOT THE CASE IN REAL LIFE, HOWEVER!!!

There are other effects which change this order The correct ranges of chemical shifts are : (ethyne) ~ 2- 3 (ethene) ~ 4- 7 (ethane) ~ O -2

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NMR Lecture 4

14.55 April 12. 1999

Chemical shifts of olefinic protons

The effect of substitution for olefmic protons have been investigated and a fonnula proposed : 3H = 5.25 + Zgem+ Zcis + Z trans Where Zgem.Zcis and ~s are the contributions (in ppm) for the substituent at that position.

RCIS .H
", C'

/ :c

/ Rtrans

"
Rgem

1'"-'-

ICOOH ! 0.97 1.41 0.71 ! Some strongly electronwithdrawing groups (OR, NR, F) have a strong deshielding effect if it is in the gemmal position (bonded to the samecarbon as the proton) due to inductive effects The same group has a rather strong shielding effect if it is in cis or trans position (the 'other' carbon compared to the proton) due to electron-donatingconjugative effects

Anisotropic

effects

Proton chemical shifts are greatly affected by magnetic dipoles at neighbouring atoms or groups -The magnetic dipoles alter the local magnetic field 'felt' by the proton. -In our molecule (diatomic, C-H) we get a magnetic moment, ~A, due to the effect the external field BO has on electrons around atom C. We can consider ~A as a localized point dipole at the centre of atom C. Please note that 'atom C' is not a carbon atom. ~A(Z) is big, ~A(Y) is small, and ~A(X) is intermediate in the picture on the next page. -When our molecule tumbles in solution, the secondary field due to the dipole ~A will increase or decrease the local, effective, field at atom H. -We assume that our molecule tumbles freely and no position is favoured. Then we will see the avecage effect of the secondary field. -The effects on H of the induced dipole at atom C are clearly not the same for all molecular orientations. C is said to be magnetically anisotropic if at least one of the ~A(X), ~A(Y) or ~A(Z) is different from the others. If ~A(X)=~A(Y)=~A(Z) then C is said to be magnetically isotropic and C will have no effect (on average) on ~ of the H close to it. It is a through-space not transmitted via bonding electrons. efTect, it is

Page 4 115

NMR Lectw-e 4

14.55 Apri112, 1999

z
~

B
/

"'\ -'\\'1"-

I
f

1/

~~

\
\ .'

I
I
t~1

I: HI ..

-\ I -"" /L ~

i\ ~~ ""

\ ~)
z

"r"

.\

I
\

,x

\t
\ y

\
/

""-"- I

:'--. "-/"-<- I ~A(x)

J L 'I,

-"..:: ~A(z)

Here we see the effect of the secondary field on the local effective magnetic field at atom H when the molecule (and the molecular coordinate system) is tumbling in solution. (left) Berr at H is increased due to the secondary field beeing parallell to Bo (H is deshielded) (middle) The molecule's y-axis is allgned with BO and Berris increased again (H is deshielded) (right) The molecule's x-axis is aligned with BO and Berr is decreased at atom H (H is shielded)
r

-Many

chemical features (atoms, bonds, functional groups etc) give rise to anisotropic effects.

For instance, even a carbon-carbon single bond is anisotropic : + Ha 0.3 nm AO'=+0.14ppm ~ -C-C
AO' = -0.28 ppm
~

K
~
\

l... 0.3 nm..1

.'., '~ ~ Hb

"'"""

Shielding cones showing how the shielding constant is changing due to anisotropic effects. -The effect on the shielding constantis negative (leading to higher chemical shift) along the axis of the bond while the effect on the shielding constantis positive in regions perpendicular to the bond (lower chemical shift) -The nodal planes where there is no change in O'(AO'= 0) goes along the surface of a cone with a 54.7 angle ('magic angle') to the bond (1-3cos2 ~) -The C-C bond anisotropic effect is clearly seenin cyclohexane where axial and equatorial protons have a -0.5 ppm chemical shift difference. At low temperaturethe chair-chair interconversion is slow and the A between Hax and Heq can be measured

Heq

4 H2

Page 5 115

NMR LectW"e4

14.55 Apri112, 1999

On the previous page, it can be seen that it is the shielding cones of the C2-C3 and the C5-C6 bonds that produce the anisotropic effects on the axial and equatorial protons at C 1 -Heq is in the negative shielding cone of both C-C bonds, while "ax is in the positive region. -"ax is shielded by -0.5 ppm compared to Heq

a.-methoxygalactose and fi-methoxygalactose can be distinguished due to the differences in chemical shifts of the methoxyprotons
~

Other bands give rise to anisatrapic effects as weIl :

""""

+ The shielding cones for C-C single bond, C-C double bond and C-C triple bond

+ The shielding cones for the carbonyl

+ group and the nitro group

Page

6 /

15

NMR Lecture 4

14.55 April 12, 1999

Another very important contributor to anisotropic effects on chemical shifts : Ring currents
In an aromatic ring system, a circular movement of the delocalized 7t-electrons will be induced This cUn'ent will, as we saw before, create a small magnetic field opposed to the external field BO (in the center of the cicrular path) Since the aromatic ring is planar, the created field will only be created when the plane of the aromatic ring is perpendicular to Bo. As a result we have anisotropy and the chemical shiit of nearby protons will be affected

B due to circulation

of 1t-electrons

Protons located above or below the aromatic system will be in a strong shielding zone leading to upfield chemical shifts Protons located in the plane of the ring (the benzene protons) will experience a significant downfield shift due to the 'reinforced' magnetic field -The 6 protons of benzene resonate at 7.27 ppm -The olefmic protons ofcyclohexa-l,3-diene resonates at 5.86 ppm -The difference of 1.4 ppm is attributed to the ring current shift

Page 7 115

NMR Lecture 4

14.55 April 12, 1999

Another interesting example :

I-I II

r
In [16]-annulene

[16]-annulene

[18]-annulene

the 'inner' protons resonate at = 10.3 and the 'outer' protons at = 5.28 on the other hand, the 'inner' protons resonate at = -4.22 and the 'outer'

In [18]-annulene,

r--

protons at = 10.75 It is the ring current effect which is responsible for these dramatic differences in This shows that [18]-annulene is aromatic while [16]-annulene is not. According to the '4n + 2' rule the same result is predicted

The ring current shifts are very important in NMR of ONA and RNA as weIl as in the NMR of proteins -All nucleobases in RNA and ONA (A, C, G, T, U) are aromatic and give rise to ring current shifts -Adenine and guanine give rise to the strongest ring current shifts, cytosine of intermediate strength and uracil of rather low strength

r'

"

Page 8 115

NMR Lecture 4

14.55 April 12, 1999

The confonnation of ONA is stabilired by both 'vertical interactions' (stacking) and 'horizontal interactions' (hydrogen bonding)
H H~N ,~..r-< ~N-< 'N-H N H
H

CH3

Distance between two base planes: 3.4 ngstrm

C1'

o
HH

'c1'
/H

o
N

N~H
C1

N'--~>H >-N,
O

C 1

The protons of a nucleobase as well as Hl' are to a large extent within the shielding cone of the next base and experience an upfield shift Transitions of a ONA duplex ~ randoffi coil can be ffionitored by NMR. Melting curves from NMR are as precise as the 'normal' uv melting curve -When the ordered, stacked structure becomes destacked (randoffi coil) the aromatic and
(

anomeric protons are no longer in the shielding cone of the neighbouring nucleobase ~ downfield shift -In non-standard structures the (temperature dependent) ring current shifts are used to determine which nucleobases are stacked to each other

Chemical shifts of exchangeable protons etc

-Protons involved in hydrogen bonds (X-R Y) are strong ly affected by the electrical dipole field created (the R-bond is an electrostatic bond) and this leads to downfield chemical shifts (deshielding) (useful for NMR of ONA and RNA) -In general, chemical shifts of these protons are very dependent on temperature, solvent, pR etc -In samples which can form intermolecular R-bonds (R-OR, R-NH2)' the chemical shift of the R-bonding proton is very rnuch dependent on the concentration -EtOR -EtOH in EtOR (neat, many R-bonds) : in CC14 (5%, almost no R-bonds) : in CC14 (high conc., many R-bonds) : in CC14 (low conc., almost no R-bonds) : 8 = 5.3 8 = 2.5 (~8 = -2.8) 8 = 7.5 8 = 4.4 (~8 = -3.1)

"'"""

,.-,
-In

-Phenol -Phenol

samples which can form intramolecular R-bonds, the chemical shift of the R-bonding proton is on ly little dependent on the concentration -salicylaldehyde -salicylaldehyde (neat, many R-bonds) : in CC4 (5%, still many R-bonds) : 8 = 11.5 8 = 11.1 (~8 = -0.4)

Page

9 / 15

,
NMR Lecture 4

14.55 April 12, 1999

H I

NMR Lecture 4

14.55 April 12, 1999

NMR

of carbon-13
The basic theories for chemica! shifts, as weIl as for NMR in genera!, are true a!so for carbon atoms One important difference is the relative sensitivity -The nuclear spin of 13C, I, is t hut the magnetogyric ratio, y, is only 25% of y for protons

This 1eads to a 1ower sensitivity since AB = ~ 27t Another important aspect concerning sensitivity is the natural abundance -For -For
~

hydrogen atoms the natural abundance is 99.98% for lH carbon atoms the natural abundance is 1.108% for 13C compared

Taking 'Yand natural abundance into account the relative sensitivity of l3C is ~ to lH -The

low sensitivity of l3C is an advantage when looking at lH spectra. With a higher

abundance of l3C, the lH spectra would become very complicated due to lH-l3C couplings. In a l3C spectrum, all those lH-l3C couplings are observed unIess special precautions are taken -The low sensitivity is naturallya disadvantage when observing l3C

Chemical shifts of carbon atoms

One important difference to 1H is that carbons have p-electrons -Carbon -The resonances are spread out over 200 -300 ppm

local paramagnetic shielding term (~ocal ) is of great importance while

ddocal(the diamagnetic term) is ofsma11 importance contrary to the lH case

"

The chemical shifts of carbon atoms follow roughly the same order as for protons (aldehyde) > o(alkene) > (alkyne) > o(alkane) > TMS Effects of substituents on 13Cchemical shifts are similar to their effects on r H) but not identical -In CH3-I the 13Cresonancemoves upfield (to -20 ppm) contrary to the lH case. Similarly, the 'expected' large downfield shifts (from lH analogy) are not seenfor CH3-Cl and CH3-Br The substituent effect propagatesthrough more bonds for 13Cshifts than for lH -For lH chemical shifts, the substituent effect is virtually only observed for protons bonded to the a.-carbon (the carbon carrying the substituent) -For 13Cchemical shifts, substituent effects are clearly observed at the y-carbon (3 bonds), in some caseseven at carbons further away -In acyclic alkanes the effect on the - and e-carbonsare small but in cyclic compounds these effects may be large

Page11/15

NMR Lecture 4

14.55 April 12, 1999

NMR

of carbon-13
The basic theories for chemica! shifts, as weIl as for NMR in genera!, are true a!so for carbon atoms One important difference is the relative sensitivity -The nuclear spin of 13C, I, is t hut the magnetogyric ratio, y, is only 25% of y for protons

This 1eads to a 1ower sensitivity since AB = ~ 27t Another important aspect concerning sensitivity is the natural abundance -For -For
~

hydrogen atoms the natural abundance is 99.98% for lH carbon atoms the natural abundance is 1.108% for 13C compared

Taking 'Yand natural abundance into account the relative sensitivity of l3C is ~ to lH -The

low sensitivity of l3C is an advantage when looking at lH spectra. With a higher

abundance of l3C, the lH spectra would become very complicated due to lH-l3C couplings. In a l3C spectrum, all those lH-l3C couplings are observed unIess special precautions are taken -The low sensitivity is naturallya disadvantage when observing l3C

Chemical shifts of carbon atoms

One important difference to 1H is that carbons have p-electrons -Carbon -The resonances are spread out over 200 -300 ppm

local paramagnetic shielding term (~ocal ) is of great importance while

ddocal(the diamagnetic term) is ofsma11 importance contrary to the lH case

"

The chemical shifts of carbon atoms follow roughly the same order as for protons (aldehyde) > o(alkene) > (alkyne) > o(alkane) > TMS Effects of substituents on 13Cchemical shifts are similar to their effects on r H) but not identical -In CH3-I the 13Cresonancemoves upfield (to -20 ppm) contrary to the lH case. Similarly, the 'expected' large downfield shifts (from lH analogy) are not seenfor CH3-Cl and CH3-Br The substituent effect propagatesthrough more bonds for 13Cshifts than for lH -For lH chemical shifts, the substituent effect is virtually only observed for protons bonded to the a.-carbon (the carbon carrying the substituent) -For 13Cchemical shifts, substituent effects are clearly observed at the y-carbon (3 bonds), in some caseseven at carbons further away -In acyclic alkanes the effect on the - and e-carbonsare small but in cyclic compounds these effects may be large

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NMR Lecture 4

14.55 April 12, 1999

OH ICHO

48.3 31.4

10.1 0.7

-6.0 -1.9

0.3 0.8

0.2 0.5

The table shows the differences in 13Cchemical shifts (L\5c) for various l-substituted pentanes.The chemical shifts for pentane are : (C1)=(C5)=13.7 , (CV=(C4)=22.6 and (C3)=34.5 -The substituent effects on the a.- and j3-carbonsare normally deshielding substituent effect on the 'Y-carbonis shielding (the 'Y-effect) 'Y-effectis very clearly seenin substitutedcyclohexanes

,-.,

-The -The

-The 'Y-effectdependson the dihedral angle betweenthe substituent and the 'Y-carbon. The classical staggeredrotamers gauche- (-600), trans (1800)and gauche+ (+60) are normally considered as the only three possible states -In acyclic compounds the rotations around C-C bonds are generally so fast that we seean averageeffect only from the three staggeredrotamers -In cyclic compounds the rotation is restricted and there will only be two possible rotamers for a dihedral X-C-C-C (where X is an exocyclic substituent)one gauche and trans. orten one rotamer is highly preferred over the other (dependingon the nature of the endo- and exocyclic substituents) -In cyclohexane an axial substituent on the a.-carbon will be gauche to the 'Y-carbonof the ring and cause an upfield shift of 4-7 ppm for the 'Y-carbon -Accordingly, an equatorial substituent on the a.-carbon will be trans to the 'Y-carbonof the ring and cause an upfield shift of 0-3 ppm for the 'Y-carbon 33.2

~CH3 H3C
(\

ty

13

36.2

(H3C)3C

Equatorial

Axial

Page12 115

NMR Lecture4

12:02 PM April 14, 1999

-The 31p chemical shiit is also sensitive to the R-O-P-O and O-P-a-R torsional angles in a phosphodiester -The torsional angles and the bond angle not uncorrelated phenomena : Changing one structural feature is accompanied by a change in the other -This interdependence is called the 'stereoelectronic 31p effect' C

,,\\\\\c 0.."' it 0p ~ ,..0 C \\\\\\"' C \\\\\,"' gauche-0 gauche0rtt ,..0 it P-O gauche() trans

"'

f"'

In practice as weIl as from semiempirica! MO ca!culations we seethat phosphateesterswith both P-O torsions in gauche conformation (-60 or +60) resonatesseveral ppm (3-6 ppm) upfield from a phosphateester which has at least one P-O torsion in trans conformation (-180) The A(31p)arising from torsiona! changesare used to monitor DNA and RNA conformation : In a stacked conformation the ~ and a. torsions (P-O torsions) take up a gauche,gauche conformation while in a non-stacked (random coil) a!so the gauche,transand trans,trans conformers exist These conformationa! changeslead to downfield shifts of the 31 p resonanceswhen the conformation changestoward random coil which can be induced by increasing temperature

f' '-"

r,

Page

14/14