FTIR analysis of secondary structure of protein
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A brief about FTIR spectroscopy and detailed description to analyse the secondary structure of proteins by Origin 8.5 software.
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1. FTIR spectroscopy can now be used to study protein structure as it requires small protein amounts and can separate overlapping signals, avoiding issues like fluorescence.
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13th Brazilian Meeting on Organic Synthesis
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FTIR analysis of secondary structure of protein
- 1. FTIR analysisFourier Transform Infar-Red spectroscopy Scope of in food research Non-Thermal Processing Laboratory Indian Institute of Technology Kharagpur Visit us: https://www.myprotocol.net/Laboratory/
- 2. Why FTIR? Fast, less sample requirement, non-destructive, qualitative & quantitative, precise and great repeatability with many others benefits.
- 4. Why Infra-Red? • IR absorption occurs from the stretching and bending of the covalent bonds in molecules. • To be accompanied by IR absorption a stretch or bend must change the dipole moment of the molecule. • Molecules with symmetric bonds such as N2, O2, or F2 do not absorb radiation in the infrared since bond stretching does not change the dipole moment of the molecule.
- 5. Two types of vibrations on energy absorbance
- 7. FTIR system
- 8. Sample Preparation A B Pellet
- 10. FTIR in food sample analysis
- 11. Oils quality
- 13. Starch quality
- 14. How to proceed? 1. Obtain the FTIR spectrum data. 2. For quantitative analysis amount of sample as well as pallet is important. These should be same. ( around 0.01 g sample and 0.5 g pallet) 3. Get the software OMNIC or ORIGIN Pro versions. 4. Some supporting and reference literature. 5. Important steps are: Baseline correction, peak finding, height/area of peaks, hidden peaks findings, deconvolution, curve fitting, area calculations It will be demonstrated how to proceed with FTIR data for protein structure analysis (one of most difficult analysis) with ORIGIN software.
- 15. Data loading and plotting
- 16. Baseline correction Baseline correction at 1600 nm because I am trying to find out the hidden peaks in amide II band of spectrum.
- 17. Peak finding
- 18. Hidden peaks fitted curves
- 20. Conclusion • FTIR analysis is extensively used as non-destructive qualitative as well as quantitative analysis for various types o samples. • FTIR analysis can be used for oils, starch, protein, or any specific functional component present in the sample. • Care should be taken while removing background or doing baseline correction. • Proper literature should be there for assignment of spectral peaks. • Chances of interpretational error can be very high if not properly analyzed.
- 21. Thanks for your attention, amitchauhan9091@gmail.com Visit us: https://www.myprotocol.net/Laboratory/