I want to know, what's the cause that we are measuring the absorbance at different wavelength? why not the same? what would be if I used a same wavelength for all the assays related to it.
Suraiya Yeasmin: We do not select wavelengths at random. A scientific method or technique should be "Selective" to have value. We develop different analytical methods to take advantage of the fact that a compound's own physical and chemical properties (and/or when altered with reagents) will help define how energy is absorbed or transmitted through them.
Total phenolic contents (TPC) relies on a special reagent for analysis ~ 750 nm (To understand the TPC reaction method, please read up on the Folin–Ciocalteu method). Total Flavonoid Content (TFC) is also measured using a specialized reagent method which is selective for 415nm.
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Dear Suraiya Yeasmin ,
These two testing methods are related to their characteristic responses. Otherwise, I learned that not all the same reactions use the same absorbance. I measured the absorbance values of different concentration gradients under some absorbance, and fitted the standard curve, measured the correlation, and chose the wavelength that is most suitable for my sample. If you are interested in this aspect, you can also determine. If the concentration gradient of the characteristic reaction of your sample has a high correlation with the absorbance at this wavelength, you can certainly use this wavelength for measurement.
Hope this could help you.
Best regards,
Weihao
Dear Suraiya Yeasmin ,
William Letter and Weihao Meng have given you some Useful Idea I also try to
make you satisfy.
It is clear that not all Compounds have the some types of reactions with all other compounds. The difference between any too compounds have exactly the same reaction properties. They may be have properties very close to each other while with others, little or very close each other. That is why all of us try to find out best compound better for his work. Of course this is because of the difference between them in their reactions. Thus, I think this will be quite enough for me to go further. I wish you the best.
Kindly check..
https://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/uv-vis/spectrum.htm
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