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DTNB peak on HPLC

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#1 herukarwadji



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Posted 29 August 2010 - 08:06 PM

Hi all,

I need an urgent help here please. I am doing a research related to whey protein denaturation right now and tried to investigate possible use of a sulfhydryl reagent, in this case DTNB ( dithio(bis)-p-nitrobenzoate, as a compound that could protect the protein from denaturation. So, I tried to skim my protein samples with DTNB on HPLC machine to see the difference peak in terms of protecting the protein from denaturation. However, I found a sharp peak everytime I run my sample and I am sure that the sharp peak is coming from DTNB (since i've already run DTNB itself on HPLC and still showing the same sharp peak at the same retention time as well). So, my question is has anyone here have done a research related to DTNB or have tried to skim the DTNB peak on HPLC? is that sharp peak suppose to come like that or is the sharp peak is DTNB's product which is TNB (after interacting with my protein sample)? Thanks guys. Look forward to hear from u all^_^ Moi

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