Fourier Transform Infrared Spectroscopy or FTIR is a technique which is widely used for biophysical characterization. It is widely used for characterize the protein secondary structure of Biopharmaceutical quantitatively. Fourier Transform Infrared Spectroscopy is a pure instrument-based test. Also, it is able to perform quickly.
The method of the Fourier Transform Infrared Spectroscopy depends on the infrared radiation absorption by the proteins as they undertake the vibrations of the molecular. Vibration of Amide 1 is particularly sensitive to the local environment. Also, it is able to reveal the information related to the protein secondary structure. The quantitative information which is obtained from this FTIR spectroscopy is not as good as what circylar dichroism (CD) obtained.
However, the Fourier Transform Infrared Spectroscopy has some advantages for you since this technique is applicable to be used for solid protein or even solution studies. Thereby, this technique is ideal to be used for biophysical characterization of lyophilized proteins biopharmaceutical formulations. The amount of the material which is used for the analysis of the FTIR is as little as 10 microlitres with the concentrations of around 0.5mM.
The method of the Fourier Transform Infrared Spectroscopy is non-destructive. It allows for protein recovery for the characterization by employing other biophysical techniques. Also, FTIR spectroscopy is able to study the formulation. Also, it is able to reveal the denaturation and the aggregation of the protein as the result of the temperature.
How does Fourier Transform Infrared Spectroscopy work?
The work of the Fourier Transform Infrared Spectroscopy is not the same as the conventional spectrometer in which the sample have to be exposed to electromagnetic radiation and the response of this exposure is then monitored. The radiation energy is varied to meet the desired range and thereafter the response is schemed as a radiation energy function.
The method which is used by the Fourier Transform Infrared Spectroscopy is not varying the electromagnetic radiation energy. Instead, it only exposes the sample to the single radiation pulse then it measures the response of the exposure. The resulting signal which is called as the free induction decay comprises a fast decaying composite of any possible frequencies. The resonant frequencies which are resulted will be leading in the signal depending on the resonance of the sample.
The principles of the Fourier Transform Infrared Spectroscopy approach can then be compared to the musical tuning fork behavior. The exposed tuning fork to sound waves of the varying frequencies, it will be vibrating while the frequencies of the sound wave are in tune. However, this is virtually the same as the conventional spectroscopic techniques in which the frequency of the radiation is varied. The response of the sample which is exposed to the radiation pulse is a signal which contains mainly the characteristic frequencies of the sample. In short, FTIR spectroscopy is a mathematical technique which is aimed to determine the characteristic frequencies of one composite signal.
Fourier Transform Infrared Spectroscopy is typically applied for the identification of the foreign material such as fibers, residues, and particulates, bulk material compounds identification, identifications of constituents in multilayered materials, and also quantitation of contamination of various materials.
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