BY: Wasim Khalili

1D NMR
Pulse Sequence

Spectrum (EtOH)

Four periods in 2D experiment

Preparation

spin system relaxes and then excited by r.f.

Evolution (t1) chemical shifts & spin-spin couplings evolve. This is the timedomain which is incremented during a 2D experiment Mixing r.f. pulses are applied and create observable transverse magnetization

Detection (t2) observable transverse magnetization is recorded. It is usually labeled with t2

2D NMR
1. Primary 2D matrix consists of a series of FIDs.

2. A set of 1D NMR spectra is obtained by Fourier transformation with respect to t2

3. The signals of each transformation may differ in amplitude and phase. A second Fourier transformation with respect to t1 yields the final 2D matrix with frequency axes F1 and F2

1.

2.

3.

2D NMR
This is how 2d NMR graph looks like Pulse Sequence

Spectrum

2D NMR

Phases... Now you can notice the spots on this line which is called diagonal (same information from 1d NMR) But there is extra spots here which refers to the correlation between neighboring atoms

Here you can notice how 2d NMR magnifies the signal and it well appear in a 3d shape like here

Now here after 2 FT we well got this shape The dotted line refers to negative signal and the rigged line refers to the positive ones Also the no. of circles refers to the height of the signal and by that we can determine small signals from big ones

2D NMR

2D NMR
This graph describes how by changing t1 you can get this 3d signal in 2d NMR

Here are the types of 2d NMR You can see that every type record a different correlation signal I am going to describe the important ones But I want you to know that j refers to the correlation signal between tow atoms and n is how far these tow atoms are far from each other

COSY L.R. TOCSY

Relayed COSY. (with one homonuclear relayed).

NOESY. ROESY.

2D J-resolved

COSY 45. COSY 90. COSY DQF. TOCSY.

H,H- COSY Experiment

N3J

HH

First type Correlation spectroscopy ( COSY) The first and most popular two-dimension NMR experiment is the homonuclear correlation spectroscopy (COSY) sequence, which is used to identify spins which are coupled to each other. COSY spectra show two types of peaks. Diagonal peaks have the same frequency coordinate on each axis and appear along the diagonal of the plot, while cross peaks have different values for each frequency coordinate and appear off the diagonal. Diagonal peaks correspond to the peaks in a 1D-NMR experiment, while the cross peaks indicate couplings between pairs of nuclei

Cross peaks result from a phenomenon called magnetization transfer, and their presence indicates that two nuclei are coupled which have the two different chemical shifts that make up the cross peak's coordinates. Each coupling gives two symmetrical cross peaks above and below the diagonal. That is, a cross-peak occurs when there is a correlation between the signals of the spectrum along each of the two axes at these values. One can thus determine which atoms are connected to one another (within a small number of chemical bonds) by looking for cross-peaks between various signals.

TOCSY of strychnine
The second type am going to discuss is the Total correlation spectroscopy or TOCSY The TOCSY experiment is similar to the COSY experiment, in that cross peaks of coupled protons are observed. However, cross peaks are observed not only for nuclei which are directly coupled, but also between nuclei which are connected by a chain of couplings. This makes it useful for identifying the larger interconnected networks of spin couplings.

N>3J

HH

In this slide you can see the difference easily

COSY & TOCSY

NOESY of strychnine
Third type is the Nuclear Overhauser effect spectroscopy (NOESY) This method establishes correlations between nuclei which are physically close to each other regardless of whether there is a bond between them. They use the Nuclear Overhauser effect (NOE) by which nearby atoms (within about 5 ) undergo cross relaxation by a mechanism related to spinlattice relaxation.

xJ

HH

C,H Correlation by Polarization Transfer (HETCOR)

Forth type is Heteronuclear through-bond correlation methods or HETCOR Heteronuclear correlation spectroscopy gives signals based upon coupling between nuclei between two different types. Often the two nuclei are protons and another nucleus (called a "heteronucleus"). You can see here at f1 is H and in f2 is C .. not identical

1J

HC

Basic HSQC Experiment

5 .. Heteronuclear single-quantum correlation spectroscopy (HSQC) HSQC detects correlations between nuclei of two different types which are separated by one bond. This method gives one peak per pair of coupled nuclei, whose two coordinates are the chemical shifts of the two coupled atoms.

1J

HC +

JHH

Basic HMQC Experiment

6- is Heteronuclear multiple-quantum correlation spectroscopy (HMQC) which gives an identical spectrum as HSQC, but using a different method. The two methods give similar quality results for small to medium sized molecules, but HSQC is considered to be superior for larger molecules

F2 displays the doublets with the spin coupling constant 1J(C-H) in addition to the H,H spin coupling.

1J

HC +

JHH

Basic HMBC Experiment

7- is Heteronuclear multiple-bond correlation spectroscopy (HMBC) HMBC detects heteronuclear correlations over longer ranges of about 24 bonds.

Cross signals caused by 1J of C-6 and C-3 are still observable but those arising from 3J and 2J are predominant.

24J

HC