Biotechnology Assignment Help With NMR spectroscopy
This technique is powerful analytical technique, which is used to characterized organic molecules by defining carbon-hydrogen structures within molecules. There are two very common type NMR spectroscopy, which is used to characterized their organic structure: H-NMR is used to define type and number of H-atom in molecules. C-NMR to define type and number of C-atom in molecules. NMR takes energy from long wavelength radio waves, which have low energy and frequency. When this low energy wavelengths enters into the molecules, then some changes occurs and cause nuclear spin within molecules including H and C atoms. When spining occurs, then there is some magnetic field created and due to this nucleus is acted like tiny magnetic bar. Normally these tiny bars are randomly oriented in space. And the presence of magnetic field molecules are against its applied field. Maximum nuclei are oriented in the applied field due to its arrangement is lower in energy. There is very small energy differences ( < 0.1 cal ). Now there are two energy states created for protons, lower states for nucleus aline with same direction of magnetic field, and higher states for against the magnetic field. When these energy difference is match with external energy sources, then energy is absorbed and cause nucleus spin-flipping forme one to another orientations. Frequency and magnetic fields are proportional to each other.
NMR spectroscopy is preferred as 300MHz, 500MHz instruments and so on, on the basis of RF radiations, which is used for resonance. In this spectrometers use very powerful magnets to create a small and measurable energy differences between two possible states.
H-NMR: In spectrum NMR absorption shows sharp peaks. Maximum proton absorb between 0-10 ppm. The NMR absorption measured according to the position of reference peak at 0ppm on the scale because of tetramethylsilane (TMS). This compound is inert volatile compound. Which gives single peak upfield from typical NMR absorption.
C-NMR:It is easier to analyze in comparison to the H-NMR, due to the signals are not splits. Due to lack of splitting in carbon is cause low natural availability of carbon (1.1%), and due to this increase the chance of bonding of two carbon nuclei, and it is very small (0.01%). Therefor no carbon splitting is occurs. Peak of carbon is singlet, and carbon spectrum provides maximum structural informations, number of signals observed and chemical shift of the signals.
Application of NMR Spectroscopy: It is used in a) Solution culture, b) finding molecular dynamics, c) Determining and residual structures of protein folding, d) Determining ionizations states, e) Screening and designing drugs, f) metabolic analysis, chemical analysis etc.
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