Hello everyone! I have two questions that I'd like to discuss.
First of all, why do ABTS concentrate once is diluted? I mean, the basic protocol says that ABTS must be diluted in PBS to an absorbance of 0.7 approximately to make the reading, and in the lab we noted once a couple hours later that the absorbance increased slightly (0.8 and little more). I suppose that is because ABTS is an oxidant agent, and if it is not stored in vaccum to avoid oxidation reactions ABTS should oxidaze more and then the absorbance should increase. Maybe I be correct?
And my other quest. I been having problems with the calculations for total phenolic content from the Folin-Ciocalteau method, and I need to know why some researchers consider the intercept from their calibration curve, and others don't. Can someone help me?
Thanks in advance for all the answers!
Concentrated ABTS gives a very dark color. The spectrophotometer can accurately measure absorbance over a limited range and fail to abide by the Lambert-Beer law over that range. Therefore, absorbance is taken in a condition that gives the best response for which ABTS is diluted to an Abs of 0.7.
You can calculate the phenolic content from the standard curve using the equation:
y = mx + c.
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