NMR SPECTROSCOPY
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Transcript of NMR SPECTROSCOPY
RRMM
SSBB
NMR SPECTROSCOPYNMR SPECTROSCOPY
JM FranconiJM Franconi
RRMM
SSBB MR SPECTROCOPY
In a spectrum, each signal represent the resonance frequencyof a nucleus in a specific environment
Principle of FT NMR spectroscopy
Bo is fixed
a RF pulse irradiate a broad range of frequency
t
RFirradiation
FID spectrum
sampling
f
FT
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Non selective RF pulse
In spectroscopy the RF pulse used is < 100 microsecond
RF rectangular sinc
B1 > 2F
F frequency range
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RF pulse Magnetisation evolution
Bo
MMz
MxyO
B1
During RF pulse M rotateAround x direction with an angle:
x
y
z
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spectrum parameter chemical shift
f1 f2 f3 f4
A same nucleus can have different resonance frequency
f
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chemical shift phenomenon
Bo
H1
O
Bo1 Bo2
H2Bo1
Bo
Bo2
C
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chemical shift measurement
f
reference
6
10ref
refi
i
ref i
DiamagneticHigh fieldLow electronégativity
paramagneticlow fieldhigh electronégativity
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J coupling
J coupling is an interaction between close nuclei
This interaction is independant of Bo field
It does not exit on one isolated molecule
f fA fB
JAB
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J scalar coupling
H1
C C
H2
50%(+1/2)
50%(-1/2)
H1 is influenced by the 2 H2 sites
Jcoupling is transported by chemical bond
f1 f1+dBH2 f1- dBH2
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J dipolar coupling
Coupling is also possible directly through space
H1 H2
1/r6
r
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J coupling
The Nuclei magneticaly equivalent have the same chemicalshift. They do not show any coupling effect
C
CHa
HbHc
fHa = fHb = fHc
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J coupling
Rules for multiplicity determination
A I nucleus coupled to n spin S give a signal with:
2.n.S + 1 componants
The componants relative intensity are given by a binomialDistribution.
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Signal area
NMR spectroscopy is a quantitative method
f
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spectrum quality
Concept of digital resolution
Digital resolution
True NMR resolution
At
DW
FT
Time domain Frequency domain
R = 1/AT
t
T2*
f
TR=1/T2*
CONDITION: TR > DR
DR
ft
AT = N.DW
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Sampling rules
DW
F=1/Dw
f2
f2
f1
f1f2
Application of Nquist theorem
2. F. Aq = N
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Sensitivity
3
ofy Sensitivit ofy Sensitivit
H
XaHX
-The gyromagnetic ratio and the natural isotopic abundancedetermine the NMR signal available
- the nucleus sensitivity is proportional to Bo2 and
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concept of population and coherence
E1
E2
Mz
population
E2
Cohérence=transition
Mz
E1
MR signal
RF
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Polarisation transfer (NOE) C
H
+1/2
-1/2
Niveau H1
+1/2
+1/2
Niveau C13
2 independant level of energy
C
H
NO COUPLING
+1/2+1/2
-1/2-1/2
-1/2+1/2
+1/2-1/2
Transition H1
Transition C13
COUPLING
system of 4 levels of energy
Longdistance
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Polarisation transfer (NOE)
H1 transition
C13 transition
POLARISATION
dH+dC
dH-dC
-dH-dC
H1 TRANSITION SATURATION
-dH+dC
-dH+dC
+dH-dC
-dH-dC
+5
-3
ff
dH = 4.dC
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SEQUENCESEQUENCE
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SEQUENCESEQUENCESPIN ECHO SEQUENCE
90°180°
te/2
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SEQUENCESEQUENCE
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SEQUENCESEQUENCE
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DEMI TOUR!
SEQUENCESEQUENCE
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SEQUENCESEQUENCE
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ECHO = coherence recovery
SEQUENCESEQUENCE
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SEQUENCE SPIN ECHOSEQUENCE SPIN ECHO
FIDFIDECHO DE SPINECHO DE SPIN
T2T2T2*T2*
Mxy
t
90 o 180o
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RF
GS
GP
GR
DAC on
TE TR
CONCEPT OF PULSE SEQUENCE
SPIN ECHO SEQUENCE
90°180°
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NMR SPECTROSCOPY APPLICATION
-Chemical structure determination (protein, ADN,…)
-Quantitative measurement(isotopic enhancement determination)
food and beverage quality control
-Spectroscopy in vivo (metabolism caracterisation)