Hi,
Most antioxidant assay for ABTS are done via spectrophotometric method.
I am still a noob in all this antioxidant assay. But it should be fine to quantify it using HPLC?
Is the below method right?
1) Mix sample + ABTS solution
2) Let it react in the dark for 7 minutes
3) Filter the solution with 0.45um.
4) Inject into HPLC. Get reading at sample retention time (example 8 minutes) at 734nm (for ABTS).
Appreciate if anyone of you can give feedback.
Definitely NO! You still use a UV-vis detector in HPLC. In ABTS assay you are not measuring ABTS itself, which is colorless. A green colored species is one-electron oxidized ABTS, commonly abbreviated as ABTS+., which has a very complex chemistry. Who knows what happens with ABTS+. during HPLC analysis?
Finally, the main requirements for all antioxidant assays are the simplicity and cost. While you want to make the assay very complex and expensive.
Thank you for the reply Yurii. As I want to quantity my sample antioxidant activity, I thought HPLC might be more sensitive and accurate, as I will take the reading at the sample retention time than direct spectrophotometric method.
So I see that most Antioxidant assays are done via spectro as it simplier and cheaper.
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