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By
MUHAMMAD AKRAM
PhD Chemistry
Worthy Teacher : Dr. UZMA YUNUS
(Associate Professor)
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Introduction of 31P NMR Spectroscopy
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Introduction of 31P NMR Spectroscopy
Nuclei with I > 1/2 are called quadrupolar nuclei. They have a non-
spherical charge distribution, and rise to non-spherical
electric and magnetic fields. They have a quadruple moment.
Table 1.1 A quick guide as to whether an isotope has zero, half-integer, or integer nuclear spin
I Atomic Mass Atomic Number Nuclei
Half-integer Odd Odd 1 31 P()
1H(), 15
From Table, it can be seen that the magnetogyric ratio for phosphorus is
about 2.5 times smaller than that for hydrogen.
The required radiofrequency is therefore also about 2.5 times smaller than that of
the spectrometer. For a 400 MHz NMR spectrometer, that would calculate to
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approximately 161 MHz
Standard in 31P NMR Spectroscopy
7
.
Chemical shift range of different type of phosphorus compounds
200 150 100 50 0 -50 -100
RPH2
R2PH
R3P
R2PX
P(NR)3
P(OR)3
P(SR)3
PX3
P(OR)2X
CPX2
CPXN
O=PX3
O=P(OR)X2
O=P(OR)2X
O=P(OR)3
S=P(OR)3
O=P(R)(OR)
S=P(R))(OR)
O=P(R)(OH)
(RO)2POH
200 150 100 50 0 -50 -100
( ppm) 8
Chemical Shift Values in 31P NMR Spectroscopy
The upfield end in 31P NMR is due to white phosphorus P4 at P= 527 to 488
ppm, depending on solvent and water content of the sample.
Chemical shift values of Phosphorus resonance also dependence upon the bond
angle, electronegativity of the substituent's and the p-bonding character of the
substituent's .
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Dependence of 31P-NMR chemical shifts on the number of hydrogen substituent's
Tertiary phosphanes (PMe3 , PEt3, PPrn3 , PBun3, and PBut 3 ) showing the continuous
downfield shift as the length of the alkyl chain increases.
The last phosphane in the series, PBut3 , should show an additional contribution for steric
effect. The actual chemical shift values are P = 62 ppm, P = 20 ppm, P = 33 ppm,
P =33 ppm, and P =63ppm, meaning that from going to methyl to n-propyl, the chemical
shift difference is = 13 ppm upfield, and not downfield as expected.
Chemical shift difference between n-propyl and n-butyl ( = 0 ppm), PBut 3,is
=83ppm downfield from PEt3 , and thus in the expected region, if one takes the
observed chemical shift difference between PMe3 and PEt3 ( = 42 ppm downfield) as
representative 10
The 31P chemical shift range for phosphorus
At the upfield end, white phosphorus P4 resonates at P = 488 ppm. This is readily
explained by an electron rich cluster structure with P-P-P angles of 60.
Coordination to a transition metal causes a reduction in electron density on phosphorus, as
a P4 lone pair is used to create a donor bond.
Coordinating the phosphane PMe3 to a transition metal again results in the expected
downfield shift of = 107 ppm, since the phosphorus lone pair is utilized for the donor
bond, resulting in a decrease of electron density on phosphorus.
However, when we form covalent bonds between phosphorus and the metal, the picture
changes. Looking at [Cp2 Hf(PCy2 )2 ], we see two different phosphorus resonances at
P = 270.2 ppm and d P = 15.3 ppm, respectively.
We note that the two phosphorus atoms are trigonalplanar and tetrahedrally coordinated,
and suspect that they are part of a P = Hf double and a P-Hf single bond, respectively.
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31P-NMR chemical shift values for phosphorus carbon multiple bonds
Normally five membered heteroaromatic ring systems ( i.e. pyrrole ) are electron rich, and the corresponding
six membered (pyridine) ones are electron poor. The 1,3,5-triphosphabenzene resonates at P = 232.6 ppm,
considerably downfield from the benzazaphosphole at d P = 69.8 ppm, as expected.
Interestingly, the 3,4 diphosphinidene-cyclobutene, featuring a conjugated, non-aromatic -electron system with
resonance structures, has a P chemical shift of P = 147.0 ppm midway between the two. The analog to
1,3,5-triphosphabenzene is stable, and shows P resonances at P = 93.6 and 336.8 ppm.
These 31P chemical shifts are evidence for the folded roof-like structure of the compound, featuring a
tertiary phosphane phosphorus atom at P = 93.6 ppm, and an isolated P=C at P = 336.8ppm, respectively.
In 13C-NMR, chemical shift of benzene at P = 128 ppm with that of the cyclopentadienide ligand Cp at
P = 118136 ppm, depending on the cationic metal that it binds to. In 31P-NMR, 1,3,5-triphosphabenzene with a P
resonate at P = 232.6 ppm, and 1,3,6 triphosphafulvenide anion, the relevant P atoms resonating at P = 216, 223 ppm.
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The influence of coordination to a Lewis acid on the 31P chemical shift
Two phosphanes for comparison, PMe3 and PPh3, and note that the downfield shift for PPh3
upon coordination ranges from = 2140 ppm, and that for PMe3 is in an even narrower
band of = 4660 ppm.
In each case, the actual downfield shift depends on the Lewis base. Factors are the geometry
around the metal (cis or trans), and the substituent's on phosphorus (P+ or PPh2 + ).
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The influence of the halogen on the phosphorus resonance of the phosphorus trihalide
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The coupling constants in 31P-NMR spectroscopy
Generally larger in magnitude than those in 1 H- or 13C-NMR spectroscopy, but they are
governed by the same principles. 1 J coupling constants of 1000 Hz are readily observed.
As with the more common nuclei, 1 H and 13C, the phosphorus coupling constants
decrease with an increase of bonds between the coupling nuclei.
However, it is frequently observed that 3 JPX coupling constants are larger than 2 JPX
coupling constants. In the following we will examine the trends in the coupling
constants of phosphorus with the more common nuclei 1 H, 13C, and 31P.
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Phosphorus coupling to Hydrogen
As with other nuclei, n JPH coupling constants have a tendency to decrease when the value of n increases.
Although 1 JPH values are frequently in the range of 4001000 Hz, the coupling constants drop rapidly
and are detectable for 4 JPH only in special circumstances.
JPH Coupling Constants value for 1 JPH increases with decreasing electron density on phosphorus. In the
series PH4 + , PH3 , and PH2 , the 1 JPH coupling constants are 547 Hz, 189 Hz, and 139 Hz, respectively.
It appears that the influence of electron lone pairs on the 1 JPH coupling constant becomes smaller with
each lone pair. 1 JPH for PH4 + /PH3 is 358 Hz, whereas for PH3 /PH2 , 1 JPH is only 50 Hz.
When the phosphorus binds to a -acceptor, such as a transition metal or a main group Lewis acid. As a
result, the electron density on P drops sharply and the 1 JPH coupling constant increases accordingly.
A good example is with PH3 , which has a 1 JPH =189 Hz as a free ligand, but a significantly larger one in
its complexes: 307 Hz in [Cr(CO)4 (PBu3 )(PH3 )], and 366 Hz in Me3 BPH3 , respectively.
Note: PF2 H (1 JPH = 181.7 Hz) has a smaller 1 JPH coupling constant than PH3 (1 JPH = 189 Hz) despite the
greater electronegativity of fluorine compared to hydrogen. The reason, of course is -back-bonding
(hyperconjugation) observed in P-F, but not in P-H bonds, decreasing the effective electronegativity of F
towards P .
Similarily, the 1 JPH coupling constant increases with an increasing sum of the electronegativities of the
substituent's on phosphorus.: H < alkyl < aryl < OR < OPh. Note: The 1 JPH coupling constant increases
with decreasing steric bulk of the substituent's on phosphorus
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Phosphorus coupling to Hydrogen
In 2 JPH (H-A1-P) and 3 JPH ( H-A1-A1-P) commonly feature is a minimum of
the coupling constant around = 90 with maxima at = 0 and = 180,
respectively. Typically, the maximum at = 180 is larger than at = 0.
The relationship between the magnitudes of 2 JPH (gem) and 3 JPH (cis and trans)
coupling constants is similar to that observed in JHH coupling constants.
3J 3 2
PH (trans) is frequently larger than JPH (cis), but JPH (gem) can be larger than
3J
PH (trans).
The electron density on phosphorus has a pronounced effect on 2 JPH (gem) and
3J
PH (cis). Coordination to a transition metal markedly increases both, whereas
electronegative substituent's on phosphorus effect only 3 JPH (cis) in a variable
manner.
The effect on 3 JPH (trans) is small, but nonetheless significant. The coupling
constants for 3 JPH (trans) are typically 1030 Hz, 2 JPH (gem) up to 25 Hz, and
3J
PH (cis) up to 15 Hz. Without electronegative substituent's on phosphorus, the
values for 2 JPH (gem) and 3 JPH (cis) remain well under 10 Hz.
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Phosphorus coupling to Hydrogen
2,6,7-trioxa-1,4-
dicyclo[2.2.2.]Octane
Phosphorus shifts at 90.0 ppm for P
and -67.0 ppm for P
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Phosphorus coupling to Hydrogen
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Coupling to Carbon
(PhCH2 ) 2 PPh PhCH2 (Ph)2 P Ph3 P Ph4 P+ Ph3 P = O
The coupling of phosphorus to carbon follows the general trends observed with other
nuclei. The magnitude of n J PC is dependent on the oxidation state and the
coordination number of phosphorus .The electronegativity of substituent's on
phosphorus also plays an important role.
The magnitude of 1JPC increases dramatically upon loss of the lone pair on phosphorus.
3J 2
PC becomes larger than JPC upon loss of the lone pair on phosphorus. For methyl
phosphorus derivatives, the 1JPC coupling constant for compounds with phosphorus in the
oxidation state +III is generally found to be small or even negative, whereas the
corresponding 1 JPC coupling constant for phosphorus compounds where the phosphorus is in
the oxidation state +V is usually substantially larger and positive.
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Coupling to Carbon
n JPC 1J
PC
2J 3J
PC PC
1J
PC
3J
PC 2J
PC
Upon coordination to transition metals, the nJPC coupling constants in trialkyl phosphanes are
influenced differently. The 1JPC coupling constant increases substantially, the 2JPC coupling
constant decreases substantially, and the 3JPC coupling constant increases slightly. A typical
example is shown in Fig.
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Coupling to Hydrogen and Carbon
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Coupling to Phosphorus
JPP in organ phosphorus compounds cover a wide range of values , where as P2 Me4 1JPP = 180 Hz, this value drops to
1 J = 290 Hz for ButMePPMeBut , 1 J = 318 Hz for Me PPBut , and as low as 1 J = 451 Hz for P But .
PP PP 2 2 PP 2 4
The corresponding doubly oxidized derivatives R2 (E)PP(E)R2 (E = O, S, Se) follow the same trend, but they can be
found to the left (larger coupling constants) from the non-oxidized analogues. P2 Me4 S2 has a coupling constant of
1 J = 19 Hz, and {P(S) MeBut} a coupling constant of 1 J = 109 Hz.
PP 2 PP
Finally, the monoxidized diphosphanes R2 (E)PPR2 (E = O, S, Se) have similar 1 JPP coupling constants to the
diphosphanes, indicating that one PALP is sufficient for the magnitude of the 1 JPP coupling constant. Note: Steric
crowding results in a decrease of the 1 JPP coupling constant in diphosphanes. Note: Lower electron density results in
an increase of 1 JPP constants in diphosphanes.
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Spectra of some Phosphorus compounds
The energy supply in cells is provided by molecules that contain phosphorus. Especially PCr
(Phosphocreatine) and ATP (Adenosine-Tri-Phosphate) carry the energy needed for most
processes in the brain and other organs. Magnetic Resonance Spectroscopy has the potential
to quantitatively detect these substances in living subjects noninvasively.
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Spectra of some Phosphorus compounds
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Pt Coupling to Phosphorus
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NMR Spectra of some other Nuclei having Phosphorus
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Applications: Monitoring the Reaction
-4.40 ppm
Verkade, G.J.; Quin D.L. Phosphorus -31-NMR Spectroscopy in seterochemical analysis. 1987. 30
Applications
Reaction pathway for
the conversion of
methylphosphonate to
PRcP. The proteins
PhnG, PhnH, PhnI,
PhnJ, PhnL and PhnM
are required for this
transformation. The 31P NMR spectra of the reaction products catalysed
role of PhnK is by PhnI, PhnM and PhnJ. a, RPnTP from the reaction
unknown. PhnGHL of methylphosphonate and MgATP catalysed by PhnI
in the presence of PhnG, PhnH, and PhnL at pH 8.5.
denotes PhnG, PhnH The methylphosphonyl group is labelled as 1-Pn.
and PhnL. Inset, 31P31P coupling of the triphosphate portion of
the RPnTP product (the , and phosphoryl
groups).
Johan V.; Andrea G. V. High-Resolution Characterization of Organic Phosphorus in Soil Extracts Using 2D 1 H31P NMR Correlation Spectroscopy. Environ. Sci. Technol. 32
2012, 46, 39503956
Applications
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Applications
Fig. 31P NMR spectra (202 MHz) of ATP (10 mM) in the absence and presence of receptor 1Zn2222 in D2O at 300
K. (a) [1Zn2222] = 0. (b) [1Zn2222] = 0.5 equiv. (c) [1Zn2222] = 1 equiv. (d) [1Zn2222] = 2 equiv.
Possible binding modes for 1Zn2222 with PPi and ATP suggested by31P NMR data.
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Applications
Enlarged portions of in vivo
proton-decoupled 31P-NMR
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Applications
A normal spectra of a forearm human muscle
displaying 6 pics and their variation during a rest -
exercise -recovery protocol. The major peaks of the
Phosphorus-MR spectrum are: phosphocreatine
(PCr), inorganic phosphates (Pi), phosphodiesters
(PDE), and the three peaks (, , ) of ATP. The
chemical shift (in parts per million [ppm]) of the Pi
is used for calculation of intracellular, cytosolic pH.
Note the decrease of PCr concentration and the
increase of Pi concentration during exercise.
Mitochondrial Disorders.
Genetic and biochemical
heterogeneity. A majority of
these diseases include
neurological symptoms
besides.
Glycolytic Defects. painful
cramps, contractures, and with
progressive weakness.
Mattei J.P.; Bendahan D .; Cozzone P. P-31 Magnetic Resonance Spectroscopy. A tool for diagnostic purposes and pathophysiological insights in muscle diseases. Reumatismo, 2004; 56(1):9-14
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Applications
In order to estimate the pseudo first-order forward rate constant (kf) of the CK reaction, we
need to measure the phosphocreatine (PCr) signal while we saturate the -adenosine
triphosphate (-ATP) resonance for different durations (i.e. the progressive ST experiment)
Assuming a two-pool exchange system between PCr and -ATP and complete saturation of -
ATP, we can estimate the exchange rate between the two metabolites by solving the modified,
for chemical exchange, Bloch equation . To confirm the efficiency of our saturation pulses,
we acquired unlocalized 31P spectra in the entire volume of the lower leg muscles in all of our
volunteers.
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Prodromos P. ; Ding X.; Gregory C.; Ravinder R. Three-dimensional Saturation Transfer31P-MRI in Muscles of the Lower Leg at 3.0 T Scientific Reports. 2014
Applications