lecture #2 pulsed nmr experiments introducing the chemical shift problems, problems, problems……
TRANSCRIPT
Lecture #2
Pulsed NMR experiments
Introducing the Chemical Shift
Problems, Problems, Problems……
RF Pulses and NMR Experiments
• Until the mid 1970’s all NMR spectrometers worked by shining a RF freq on sample and slowly scanning the magnetic field
• One passage took ca. 12min• Lots of those minutes were expended on
scanning through regions with only empty baseline
RF Pulses and NMR
• Recall the thinking on Fourier Analysis• Measure all frequencies at once (better use of time)
deconvolute later• How to do this? Hold magnet constant and irrad with a
short pulse that contains all the relevant RF frequencies• All the magnetic moments oscillating at characteristic RF
freq should come into resonance (absorb energy)• In returning to equilibrium, they should release energy,
oscillating at their proper frequencies• Oscillating magnets should induce an AC voltage in a
nearby coil• Thank you Professor R. Ernst
Let’s look at this as a picture…Z
X
Y
M
x
y
z
Recall that this is vector resultant from individuals all oscillating around Z at Larmor frequency
RF pulse with whole range of frequencies
Absorbs energy
Destroys Boltzmann excess
M Still has precessional torque around z (field) axis
The RF Pulse Generates Phase Coherence
X
Y
Z
X
y
x-Pulse tips the ensemble of M down to the y axis of the x-y plane.
Another effect is to start them precessing about z at the same time point, therefore with the same phase. This is called coherence
Free Precession of M about z…
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
time
But, M shows a Damped Oscillation….
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
time
Because M is spiraling back to the Boltzmann equilibrium
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
x
y
z
time
And we are left to see the oscillation of the projection that remains in x,y plane
Pictoral of how this becomes the NMR experiment
H0
If we apply a second field at the same frequency, but from a different direction, the same kind of torque is experienced by I. This amounts to perturbing the population equilibrium
And now a miracle occurs..This “perturbation” acts like any other momentum vector in the H0 field and begins to precess about z (what frequency?)
The secret is creation of a phase-coherence that starts off the individual vectors comprising Iy having the same phase
This induced precession can be detected by contriving to have a sensing coil at right angle to H0. Coil produces voltage at same periodicity.
H0Induction!!!
Our Friend, the Chemical Shift
The Chemical Shift
• The Chemical shift makes NMR useful in Chemistry (they
named it after us)
• Arises from the electrons surrounding our nuclei, responding to
a magnetic field.
• Induced circulation of electrons, Lenz’s law; this circulation
generates a small magnetic field opposed to H0
• The small negative field diminishes the H0 experienced by a
nucleus. This differentiates sites, based on chemical nature
• Effect grows directly proportional to H0
Some History
W.G. Proctor, F.C Yu; Physical Review, 77, 717 (1950)
W.C. Dickinson; Physical Review, 77, 736 (1950)
An early example, revealing the sorting out by chemical environment, and response proportional to number of hydrogen atoms
Theory Underlying the Chemical Shift
• Bulk Susceptibility is corrected for by internal shift reference
• Shielding by electron cloud is experienced at the nucleus
• Induced circulation of electrons such that a “current flow” is set up, generating a magnetic field counter to H0 (Lenz’s Law)
• Implies that if we know about the electron cloud distribution, we could Predict chemical shifts
• Predicts direct proportionality of the chemical shift (when expressed in Hz) to the applied field. The ppm scale normalizes out this effect. This means that a 3ppm shift on a 100 MHz instrument is 300 Hz from TMS. The same 3ppm signal on a 500 MHz instrument is 1500 Hz from TMS.
A Picture of this…
A Vector Picture Chemical shift is the ultimate precessional frequency of the vector component of M in the plane perpendicular to H0
Precesses at a frequency
This is in units of (radians)/sec
At some time, has distinct angle and as a vector in x,y can be resolved into x, y components.
The receiver works by counting how many times this electric vector whizzes past in a unit of time
X
Y
H0(Z)t
X
Y
H0(Z)
After a pulse…
t
Free Precession, Rotating Frames and the Chemical Shift
•Our vector picture can help
Rotates at H0 MHz
Stands Still!
What if we could contrive to measure
once every H0 seconds? Strobe effect
Is The Rotating Frame
Now, more than one chemical shift wil move with just a difference from H0
Don’t have to distinguish 25000002 from 25000005 Hz, but 2 cf. 5
Imagine a “blinking eyeball”, (strobe effect) blinks at Larmor frequency……
What would our “blinking eyeball” receiver in the X,Y plane see, watching
this Vector over time?
Time
Vol
tage
The strobe effect cancels out the Larmor (MHz) frequency, leaving behind the chemical shift frequency
Seems to “die away…
Because the nuclear spin is also spiraling back to the Boltzmann equilibrium, leaving less “signal” in the x,y plane. (The red vector is “seen” by the x,y plane detector)
“Practical” Theory
• The real triumph of the shift theory is in its relationship to electronegativity and hybridization and easy prediction of trends based on qualitative notions from structural theory.
• Withdrawing electron density diminishes the screening ability of the electron cloud and the nucleus goes to higher field.
• Feeding in electron density sends nucleus to lower field.• “Moving” electrons have some real consequences on
nearby chemical shifts.
Defining Shift Scales
Some Useful Shift Ranges
0246810
Acids, H-bonded OH aromatic
alkenyl
CH2 allylic, acetylenic, to carbonyl
CHX CHR-Omethylene methyl
CH=O
OH, br, variable, SH sh. ca 1.5
CHR-N
ppm from TMS
1H
04080120160200
C=O aldehyde, ketone C=O,
acid
C=O, amide
CH aromatic, alkene
heteroaromatic
CR(-O)-O
C-subs aromatic, alkene
Alkyl N-subs
O-subs
13C
O-subs aromatic, alkene
15N Chemical Shift Ranges
Taken from G. Levy in Concepts in Magnetic Resonance, 6, p 338 (1994) Shifts vs. NH3(liq) Subtract 380.4 to scale to nitromethane=0.0
See also G.C. Levy and R.L. Lichter, “15N Nuclear Magnetic Resonance Spectroscopy”, J. Wiley and Sons (1979); also a massive collection of tabulated data in NMR: Principles and Applications 18, in the Chemistry Library at Temple
Chemical Shifts Sense and Report on Structure
• 13C Shift is sensitive to branching, e.g.branched hydrocarbons
Kth Carbon =
€
Bs + DM2
4
∑ ASM +γ S NK 3 + Δs NK 4
NKP = number of carbons P bonds away
DM = number of carbons bonded to Kth carbon,
with M attached carbons
S = number of carbons bonded to Kth carbon
• Sterics, electronegativity, strain, hybridization all contribute to the observed value for chemical shift
Electronegativity and substituent Shift Effects
• More Reliable in 13C
• Best used as general predictive for trends. Evaluate for consistency
• Here probably separation into resonance, inductive would help
• Changes in hybridization
• Other contributor is steric compression effect (branching?), shielding effect
Electronegativity Effects on 13C
Shifts
No Surprises Here
Chemical Shifts; Predicting and Additivity Rules
• Sometimes prediction works
• Better for carbon than for proton
• Multiple substitution can lead to push-pull deviations due to resonance, etc.
• Protons have larger relative effects on them from anisotropic neighboring fields mostly because the range of the shift domain is so small.
• Best efforts are in interpolation schemes based on mapping of assigned shifts in chemical-bond space
• The good news is that relevant model compounds are really effective in predictive value
Anisotropic Shielding Near Electrons
H
Circulating Electron Cloud responding to H0
Increases the total .field felt at H by ca
1.5 ppm
Deshielding Region
Shielding Region
Applied H Field0
Induced Current Induced Magnetic Field
Pronounced effect for aromatic, in line with e circulation
Other Anisotropic Shielding Cones Nitriles, acetylenes
isonitriles
Carbonyl, alkene +
+
++
+
++
+
++
++
In planedeshielded
Above, below planeshielded
OC
Polarized effect
Small pos
• Effects are ca. 2 ppm at most.
• Most Significant when a nucleus is fixed in geometry with respect to the neighboring field.
Best description is in
L.M. Jackman, S. Sternhell, Applications of Nuclear Magnetic Resonance Spectroscopy in Organic Chemistry, Pergamon Press, (1969) ch.2
Examples of Anisotropic Shielding
OH
O
H
1.27 1.67
H H
-.7 (to higher field)
Shielding by cyclopropyl ring
Use for both assigning signals, and interpreting the structure
Powerful Application in Study of Aromaticity
H
H
0.2 ppm
8.9 ppm
The shift anisotropy cone from the aromatic ring current requires a deshielding region outside and a shielding on the inside. An excellent review of the use of this probe is found in W. LeNoble, Highlights of Organic Chemistry, Marcel Dekker , (1974) ch. 9
L.M Jackman, F Sondheimer, A.A Bothner-By, Y. Gaoni, R. Wolovsky, Y. Amiel, D.A. Ben-Efraim, J Amer. Chem Soc. 84, 4307 (1962)
18-AnnuleneAlso for porphyrins, etc
9.2 ppm
-3 ppm
Deshielding from the C-OH bond
• Here is a dramatic example
H HOH
H
3.88 0.55
Isotropic vs. Anisotropic Chemical Shifts
• Anisotropic has shifts differ according to the angle of the molecule compared to H0
• Solids
• Preserves all the information about the interaction
• Isotropic has motions fast enough to average the chemical shift, and remove the dependency on the angle
• Liquids• Simple enough to understand
because some information is lostPowder Patterns and Chemical
Shift Anisotropy
Pattern on Right is onlyfor axial shift tensors
Typically these patternsare 10’s of ppm wide
σA ( )is the normal liquid or“isotropic” shift
To be effective at wiping out these patterns the spinning must occur at a rate comparable to thelinewidth.
3-8 This amounts to spinning at KHz for NMR field strengths commonly. .used Spin rate must be stable
Imagine frozen cyclopentadiene. Its grid has angle w.r.t. magnetic field
Different interaction of electrons with H0--Different chemical shifts! H0
In Liquids motions Averages out the Chemical Shift
The same average shift for the same chemical-electronic environment
Here the CH2s are all the same, as are all the CH next to the methylene, etc.
How do we know what “same” means?
H0
Magnetic field
How many signals do we Expect in an NMR Spectrum?
• The Chemical shift implies that we see (potentially) a different signal for every different chemical environment.
• Chemical environment here is the electronic structure (electrons, hybridization, charge, polarizability etc.) These are all things able to be predicted to some extent by theory.
• What do we mean by “different”?
When are NMR signals from a nucleus Equivalent?
• Isochronous (same frequency)• Only if the atoms are exchanged by any* symmetry operation
for the molecule. Example C2, C6
• Could be made equivalent in rapid chemical process, e.g rotation, exchange
• True always in achiral solvents• *Atoms only exchanged by mirror plane symmetry are
enantiotopic. Non-equivalent in chiral solvents• For molecules as units, similarly, enantiomers are only
distingushed with different shifts in chiral solvents. Diastereomers, like other isomers have different shifts regardless of solvent.
Are two signals equivalent, or not?
Some definitions and examples…
Homotopic
Enantiotopic
Diastereotipic
Relationship Example: 2CH3 Appearance
CH3CH3
CH3CH3
OH
CH3CH3
H
In any solvent
In chiral solvent
In any solvent
In normal solvent
A test for enantiotopic protons or 13C
Draw two structures, successively replacing A, then B.
If the two structures are enantiomers, then the signals will be enantiotopic. The carbon they are attached to is termed “prochiral”.
Basis for a lot of structure work
• Number of symmetry different positions can differ for isomeric possibilities--rule structures out
• Symmetry, Symmetry, Symmetry…..but….
Diastereotopic Signals
O
O
CH3CH3
OH
CH3
H
H
These methyl groups are not chemical shift equivalent--No matter how fast they rotate, they never see the same environment
Symmetry and
Equivalence
A Symmetry Example
HOH
H
HH
OH
H
H
H
H
OHH
H
HOH
HH
H
H
H c
c
R,S pair
Ha,b are diastereotopic and never have same chemical shift
Hc are equivalent except in chiral solvent
Ring flipping only able to distinguish at low temperature (use highest symmetry)
d
eC2
OHOH
H
HH
HH
H
H
H
a
bMeso
d
e
Ref: E. Eliel and S. Wilen, Stereochemistry of Organic Compounds, J. Wiley & Sons (1994) ch. 6R. Silverstein, G.C. Bassler, T. Morrill, Spectrometeric Identification of Organic Compounds, Wiley, (3rd Ed is 1974) ch. 4
Use What We Learned about Symmetry and Chemical Shifts
How many 13Carbon signals would we predict for these compounds?
Motion has an EffectTwo protons or carbons that are technically not exchanged by a symmetry operation can be nevertheless equivalent, if they are exchanged by a chemical process on a time scale faster than the NMR time scale.
Example, ring flipping of conformers; rotation of methyl groups.
H
H
H
H
H
H
H
HH
H O
ax
eq
ax, eq. H not symmetry equivalent but you could only see the difference at low temperature
O
Ha
Hb
At room temperature, motions make it seem “flat” with Ha, Hb at same shift.
What is meant by “The NMR Time Scale”?
•Imagine two signals that are chemically changing their identities.
•They have chemical shifts, 1, 2
•These shifts are also separated by a given number of Hz; (=1-2)
•Remember, that Hz has units of 1/sec.
•The chemical shift difference in Hz can be compared to a “chemical lifetime” or its reciprocal the reaction rate constant k. k has units of 1/sec.
•If the reaction rate k is faster than , we can only observe a signal at the average of the two chemical shifts. Intensity will be the sum.
•We can address this experimentally by making k smaller (lower the temperature) or making bigger (use a higher field NMR magnet)
•Practically, the relevant time scale for exchange here is 10s of msec.
Pictorally,
A B
An irony, samples appear “colder” w.r.t. kinetics on higher field NMR systems
Problems…
A Mini-ParadigmTabulate
Observation or Fact Inference about StructureStep 1.
Do we have enough Data? What questions do we need to address?
Molecular Weight (mass Spec)?
Inventory proton,carbon counts into shift categories, number of unique signals
Assess purity, can we “ignore” some signals?
From above, can we write down Molecular Formula?
UV chromophore?
Step 2. Tabulate Obvious features
Check the IR spectrum, and 13C NMR for Functional groups (C=O, CN, OH etc.
Mass Spectrum--Any Fragments or losses that are structurally useful? (loss of water, CO2, CH2=CH2; tropylium, acylium present?
Evaluate chromophore, from UV if available
Are there obvious 1H NMR signals by inspection? (methyls, methoxys, aromatics,
Evaluate exchangeable H from mass spec, or 1H NMR (D2O exchange)
Step 3. Start putting the pieces together
From the list of inferences in our table write out fragments that must be present.
Some must be at ends, some must be internal
Compare to molecular weight and deduce formula
Can we infer the presence of heteroatoms?
Compute DBE
Specify Fragments from 1H NMR spin patterns
Recognize that some of the parts and fragments overlap or are redundant. Tabulate this
Write down trial structures. Cross check against data.
Formulate questions. (how can I exclude ….?; how could I distinguish A from B, etc. Symmetry comes in.
Step 4. Confirmatory
Exact Mass
Comparison with known structure
Literature, data bases
“Fingerprint” available?
Inductive and Deductive Reasoning
The steps discussed frame a sort of inductive reasoning
Your knowledge of Chemistry, e.g. valence, what bonds to what, molecules you know, provide for inductive reasoning.
These two converge when you can write down a reasonable structure that agrees with all the data.