But it turned into dark blue colour on mixing. Is the reagent of dark blue color before adding the sample?
Method:
200ml of acetate buffer(pH 3.6) + 20ml TPTZ+20ml ferric chloride solution+24ml water
It is normal for the solution to be an intense blue color. You will be looking for a change in absorbance at 593nm with your spectrophotometer.
I am talking about reagent solution color before adding sample.
what is the color of reagent solution.(without adding sample)
Ideally the solution should be sort of yellow if there is no reduction of the Fe(III). Being blue before adding your sample may indicate improper pH or that the chemicals have aged too long. What is the source of your TPTZ? Are you dissolving it in 40mM HCl fresh before each experiment? Your FeCl3 solution should also be fresh at each experiment. You should also be keeping the mixture at 37oC in a water bath.
I have made my FRAP working solution with ratio of 1:1:10 and it shows a dark blue solution. Is it because my ferric chloride solution is not in orange/straw color but blueish grey once add into distilled water?
Hello @ Ron Reiserer sir..
I wanted to know that How to dilute TPTZ???
I have tried but it wasn't dilutes in 40 mM HCl solution.@
When I added Sodium acetate buffer, tptz stock solution and fecl3 hexahydrate in 10:1:1 ratio, the Mixture (FRAP reagent) showing intense blue colour. Is it normal or having some mistake
When I added sodium acetate buffer , tptz stock solution and fecl3 in 100:10:10 it become straw coloured but a colloidal solution,some white particles formed in the FRAP reagent.
This is fine or is their any mistake...
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