Hi guys, Needed a bit of help..

how do u predict the number of peaks and relative peak area for compounds given as molecular formula (i) CH3CH2CH (ii) CH3CHOHCH3 (iii) CH3CH2OCH3 I can do it for C-13 but im all confused with H NMR HELP PLEASEE!! Thanks It's very similar for the proton NMR. a) the number of peaks in the low resolution spectrum = the number of hydrogen environments (ie hydrogens that have exactly the same groups surrounding them) b) the peak area = the relative number of hydrogens in each environment If you are asked about coupling/splitting patterns in the high resolution spectrum then it's just the number of hydrogens on adjacent carbon atoms (to the carbon the hydrogen in question is connected to) +1. eg if the hydrogen is attached to a carbon with a CH3 next to it, the splitting pattern will be 3+1 = 4 = a quartet. I suggest you draw out each of the structures above and try to see which hydrogens are in identical environments to each other. Then follow the above. Good luck.

Did it for the first one (which i assume should be CH3CH2CH3 correct?

"Predict the splitting pattern that would be seen in the proton n.m.r. spectrum of the isomeric compound ClCH2CH2COOH" I got a triplet for the CH2Cl, a triplet for the CH2 it's bonded to, and a singlet for the OH group; but the mark scheme only says two triplets. Do OH groups not have peaks in NMR or is it a mistake on the mark scheme? Thank you! s there anything on the question about it being mixed with D2O (deuterium oxide), if so the OH peak disappears. If not, I can't see how you're wrong...

How are they in separate environments???????????????????? ???????????????????????????????? ? You need symmetry for them to be in the same environment. There isn't symmetry there, because of the OH group on one side compared to the ethyl, methyl and OH groups on the other side. The point is, they are equivalent, because of their relative positions to the other groups. You need to think about what each group does individually, the effects of them don't just stop at the adjacent carbon, so the OH group on the right affects both the C it's attached to, and the methylene group attached to that. It also helps to think of them in their 3D structure, and electron density in different places.
So it matters what the environment of the carbon (or group) bonded to the carbon bonded to the hydrogen as well?

the two in the middle you are pointing at are in separate environments, the carbons next to them may both have C=O groups but the next group on the right is an OH group, whereas the

one on the left has another carbon then an OH group. Unless it is exactly the same on both sides then they are separate environments am assuming that you lack basic information about the nature of NMR spectroscopy. The links I have provided go through the basics of the technique with the science behind it. Hope this is helpful.

Source(s):
http://orgchem.colorado.edu/hndbksupport http://www.chemguide.co.uk/analysis/nmrm http://www.cem.msu.edu/~reusch/VirtualTe
In the NMR spectroscopy, the atomic nuclei is absorbed radio frequency radiation under the influence of an applied magnetic field is studied. 1. It is base on the principle that the atomic nucleus which have odd mass and atomic number contain both the spin angular momentum and a magnetic moment. 2. The possible orientated spin states = 2I + 1 where I = nuclear spin quantum number. 3. The magnetic moments are randomly oriented or orientation of equal energy in the absence of magnetic field while the spins are arranged in or in opposition the alignment in the presence of external magnetic field. 4. In the presence of magnetic field, the energy level are splitted an each level gives a new magnetic quantum number. 5. The excitation of nuclei from the lower spin state to the upper spin state is done by absorption of energy. 6. This excitation produces a signal of the NMR spectrum. 7. The energy difference between the two spin states depends on the strength of external magnetic field which is generally very small. 8. The NMR spectroscopy requires strong magnetic field. 9. The NMR spectroscopy is used to determine the structure of molecules. 10. The hydrogen has three isotopes; hydrogen, deuterium, and tritium. The magnetic moment of the proton is approx = 2.7927. 11. The hydrogen atoms should give the resonance signals at the same frequency due to same magnetic moment of all protons but it is different in different compound. The difference is due to different surrounding of hydrogen atom. 12. Like in case of CH3CH2CH2OH contains three different types of hydrogen; the H in the methylene (CH2), H in the CH3 and the H in OH. 13. The compounds with more shielded proton give resonance signals at the higher field like CH4, HCl, HBr, HI etc while other give at lower field. For example in water, the resonance signal is shown at field 2.3488 T at the initial external field value 2.3487 T and 100 MHz radiation. 14. It occurs at very small increment so the frequency scales are used.

Chemical shift in proton resonance

The NMR resonance signals depend on the external magnetic field and frequency of radiation. The NMR spectrum is the graphical representation between applied radio frequency and absorption. Thus the frequency of resonance signal is known as chemical shift and denote as d.

The chemical shift is defined as the resonance frequency when standard compound is taken at 0ppm as reference. The chemical shift does not depend on frequency of spectrometer. So the chemical shift is = *Frequency of (signal reference) / frequency of spectrometer+ x 106

Shielding in H-NMR
The structural factors are the factors which cause the change in resonance frequency and then the chemical shift due to alter the value of magnetic field by nucleus. Thus the different protons show the different chemical shifts. This is the basic for the structure determination of the compounds. This includes the electro negativity, magnetic anisotropy, and hydrogen bonding. 1. Electronegativity The isolated protons show the resonance at lower value of magnetic field strength than the covalently bonded hydrogen. The protons are surrounded by negatively charged particles. When the external magnetic field (Bo) is applied then these charged particles move and a secondary magnetic field is produced. This secondary field is very stronger than applied external field. Thus the protons are shielded by electrons. The change in the experienced magnetic field of proton depend electron density. Heff = Ho-He where Ho is the spectrometer magnetic field He is secondary magnetic field. But the presence of electronegative group reduces the shielding of proton by electrons as it decreases the electro density around the proton. This results in increasing the chemical shifts of proton. Thus the presence of more electronegative group increases the deshielding and hence the chemical shift. So the descending chemical shift order is CH3FCH3Cl The order is according to the decreasing electronegativity of F,OH,Cl,Br,I etc. This inductive effect is decreased as the distance form the electronegative atom increases. Like in case of chain (CH2-CH2-CH2Br), the chemical shift is decreased form 3.40ppm (for first proton attached to Br) to 1.20 ppm for the last proton. 2. Magnetic Anisotropy

The magnetic anisotropy defined as the non uniform magnetic field. This is due to the presence of p electron system in the compounds like aromatics, alkenes etc. When the external field is applied, the p electron system interacts with the applied magnetic field.

This interaction generates induced magnetic field which is the reason for the anisotropy. It can be shielded or in deshielded form. Now the protons experience three magnetic fields; the applied magnetic field, the secondary field due to shielding of electrons and induced field due to p electron system. And this changes the chemical shift.

3. Hydrogen Bonding

It is the bond between hydrogen atom and electronegative atom. The hydrogen bonded protons show the high range of chemical shift due to high deshielding of protons. But the hydrogen bonding is also affected by acidity, temperature, concentration etc. This figure shows the chemical shifts in the presence of different groups.

The deshielding of proton is due to hydrogen bonding of the alcohol OH to the sulfoxide oxygen. The NMR spectrum of CH3Br is shown below; it shows one peak at 2.7ppm due to single type of proton which is slightly deshielded due to Br.

4. Coupling in H-NMR

Generally in NMR spectrum of protons, all the different types of protons shows as singlet but in some cases there is group of peaks are shown. The groups of peaks are formed due to the coupling between adjacent protons. It can be seen in the NMR spectra of 1,1-dichloroethane (CH3-CHCl2). The compound contains two types of hydrogen so it gives two peaks but one is in the form of doublet and other is in the quartet form at 2.1ppm and 5.9ppm respectively. The doublet peak is for -CH3 methyl group. The CH group gives two orientations with respect to applied field which split the signal of -CH3 methyl group into two peaks. While the quartet denotes for CH group which is affected by the -CH3 methyl group. It gives four magnetically different effects which splits the CH signal into quartet with intensity ratio 1:3:3:1. The signals are splitted into n+1 lines if the "n" equivalent hydrogen atoms are present on neighboring carbon atoms. This splitting is known as the coupling of protons. But the equivalent protons which have the same chemical shifts do not undergo the coupling.

5. Coupling Constant

It is expressed in terms of J as Hz unit. The coupling constant is the measurement of the interaction between pair of protons. Like in Ha-C-C-Hb then the coupling of Ha and Hb is given as Jab. The coupling of H-NMR is not always follow the (n+1) rule because of the non-equivalent nature of protons. If the proton has two types of neighboring protons then number of lines is equal to (n1 + 1) (n2 + 1). For example in 3-bromopropene or allyl bromide (Br-CH2-CH=CH2), the H on C2 has three different protons so it is coupled by three different coupling constant.

6. Solvent effect

The most common solvent for NMR spectrum is Chloroform-d as it is good solubilizing agent and unreactive character. Some other used solvent are deuterium oxide, benzene-d6, acetone-d6, and DMSO-d6 etc. the chemical shift is also depend upon the type of solvent. Some solvent shows the minor changes in the chemical shift. But the resonance spectra for C-H signals in benzene-d6 shows peak at small upfield shift while it is approx five times higher in the acetone. This is due to shielding of proton by formation of complexes with benzene ring. Similar the NMR spectrum of tert-butanol in DMSO is shifted to down field than the spectrum in dilute chloroform solution. This is because of deshielding of proton and spin-spin splitting.

Protein NMR Spectroscopy

Back to Top

Proteins play a very important role in living organism for skeleton support, movement of muscles, digestion system; defend for infection etc. they are in various shapes like round shape in hemoglobin, long shape collagen, strong spectrin-c etc. Proteins are made with amino acids which gives the specific character to protein. The protein folding state is a stable state than the unfolded state of protein. In both the states, they form various hydrogen bonds and non-ionic interactions. The folding state is more ordered and compact containing the long range interactions among the different chains of proteins. The protein folding process is quite complex because it is very difficult to know that how protein folds into correct structural forms with in tiny of seconds. So the important techniques were found to know about the complex process. One is X-ray crystallography and another is nuclear magnetic resonance spectroscopy (NMR). It is based on the identification of the observed signals in the spectra. These observed signals are correlated with the amino acid protons which gives the signals. 1. 2. 3. 4. 5. The 3D-structure of protein is determined under the magnetic field. Thus nuclei start to flip between two energy states in the specific radio frequency region. This is detected by spectrum. The proton-proton distances are determined by knowing the signal intensities. These intensities are determined by 2D NOESY, 3D 15N-NOESY-HSQC, and 3D 13C-NOESYHSQC spectra method. It depends on distance between two nuclei. If the i and j are two nuclei then the Signal intensity is NOEij ~ 1/rij6. 6. Thus the distances are measured from the spectra with use of NOE signals for standard distances which are divided in few groups. Two bounds are assigned to the all five NOE classes which are given as below. NOE class Lower bound A- Upper bound in A-

Very strong Strong Medium Weak Very weak

2.3 2.8 3.1 3.5 4.2

2.5 3.1 3.4 3.9 5

All non-sequential signals are the signals which are approx greater than 1000 in a medium-sized protein (contain 120 amino acids). These visible signals are assigned in the NOESY spectra. There is also medium range and long range NOEs. The medium range NOEs (less than five amino acids) are indicate the conformation of backbone and are used for determination of secondary structure. While the long range NOEs shows the global structure of protein and use for the tertiary structure of protein. The COSY spectrum or a HNCA-J spectrum is used for the detection of p-dihedral angle of proteins.

Tertiary Structure
The computer calculations is used to detect the distance- and torsion-angle data into a visible structure but this information is not sufficient for the proper determination of protein structure. So the determination of randomly folded starting is done by the known amino acid sequence. Then the computer program folds the structure in order to compensate the observed inter proton distance with calculated structures. The energy potential is given by known parameters. The protein molecule have tendency to take various conformation due to free rotation in chemical bonds. All the possible conformations are known as conformational space. So for determination of structure of protein by the NMR, the conformation space should be restricted. Especially computer programmes are based on two programmes for calculating a protein structure in solution which are as below; 1. Distance geometry (DG)

It is not used the Cartesian coordinate system directly. In this method, atom of each pair matrices of distance constraints from available distance constraints, torsion bond angles, and Van der Waals radii is measured. Then these calculated set of distances are projected from the n-dimensional distance space to the three-dimensional space of a coordinate system. Thus coordinates of all atoms of proteins are determined.

2. Simulated Annealing (SA)

The Cartesian coordinate system is directly used in this method. It is a molecular dynamics method. The process starts with increasing thermally mobility of atoms. This is done by heating the structure up to a high temperature in a simulation. It undergoes from many discrete cooling steps. And then converts into the energetically favorable final structure. The final structure is obtained under the influence of a force field of the constraints. Thus the NMR technique is useful for the spatial orientation of protein without the need of crystallize proteins. The process is done in the solutions of test material of protein or macromolecules. The sample can be measured in the short range of time by NMR spectroscopy. So it can be used to detect the individual molecular group movement and complete domains of proteins.