I immobilize enzyme on chitosan macrobead with Glutaraldehyde as crosslinker. I want to know how FTIR confirm immobilization? Which wavelength is related to chitosan with glutaraldeyde and which of them show immobilized enzyme?
I attach files related to FTIR
p1:chitosan
p2:chitosan+GA
p3:chitosan+GA+Enzyme
p4:free enzyme
Dear Dr. Rahimizadeh,
I was looking on your FTIR spectra and, at the first glance, I could say that only the spectrum of the enzyme is well resolved. First of all, I recommend you to check the purity of your chitosan (CS), and after that to prepare CS microbeads cross-linked with GA. To obtain clear information about the functional groups present in your materials, you have to start with pure CS, otherwise the FTIR spectra could not help you to get valuable information. That is very important because after the cross-linking of CS with GA , imine bonds result, which main absorption band is located at 1655 cm-1, i.e. in the same region with the amide I band in CS. In the spectra 1-3, neither the region characteristic for the anhydroglucose units (AGU) in CS is well solved, i.e. the peaks at 1032 cm–1 and 1074 cm–1 assigned to the skeletal vibration involving the stretching of C-O bonds in AGU are not clear evidenced.
I hope that this answer will help you to go ahead with your investigations.
hi. you have to find N-O IN 1384 cm-1
please read attached file too
good luck !.
check the attached links and I hope to be useful to you
http://www.ftirsearch.com/default2.htm
http://www.labcognition.com/web/Products/irAnalyze/tabid/126/Default.aspx
http://www.infrared-spectra.com/
Dear Dr. Rahimizadeh,
I was looking on your FTIR spectra and, at the first glance, I could say that only the spectrum of the enzyme is well resolved. First of all, I recommend you to check the purity of your chitosan (CS), and after that to prepare CS microbeads cross-linked with GA. To obtain clear information about the functional groups present in your materials, you have to start with pure CS, otherwise the FTIR spectra could not help you to get valuable information. That is very important because after the cross-linking of CS with GA , imine bonds result, which main absorption band is located at 1655 cm-1, i.e. in the same region with the amide I band in CS. In the spectra 1-3, neither the region characteristic for the anhydroglucose units (AGU) in CS is well solved, i.e. the peaks at 1032 cm–1 and 1074 cm–1 assigned to the skeletal vibration involving the stretching of C-O bonds in AGU are not clear evidenced.
I hope that this answer will help you to go ahead with your investigations.
Dear Dr. Rahimizadeh,
I agree with Dr. Dragan. It is important to start with pure chitosan. Your transformations will be easier if you start with chitosan possessing low degree of acetylation. You can also check for CH2 deformation (around 1460 cm-1) and carbonyl from aldehyde (around 1725 cm-1). Which conditions do you used to link glutaraldehyde on chitosan ?
Dear Parastou Rahimizadeh,
Try also to work with dry samples. Your spectra have intense water bands and one of them is placed in the amide region. If you want to observe changes in this region and work with water solutions you have to subtract a reference spectrum of water.
It is also necessary to control the humidity of the environment. Most of your spectra show water vapour peaks.
Good luck.
Ramon
thanks a lot Dr.Stela Dragan and Dr.Gagnan for your guidance.I activate chitosan bead with0.7% GA in28 degree for4hourse.I don;t know why I can't say chitosan crosslinke by GA as IR.but I say immobilization occur,beacause I load Enzyme on chitosan and enzymatic assay show me there are enzymes on support.
It is not possible to tell you the peaks on this message. There are several sites, Go to google search and ask for peak identification key for FTIR results. You will get many ready reckoner. Only remember that you can not identify your compound only by FTIR as it will give you functional groups only. You need to supplement it with NMR and MS data.
Thank you for important discuss. Can i get explain about -CH2 deformation or -NH deformation in ATR-FTIR spectra?
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