The working solution was prepared by taking a volume of the previous solution and diluting it in ethanol until its absorbance at 734 nm was 0.70 ± 0.02. The solution was also pre-equilibrated at 30oC before use.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C.
Antioxidant activity applying an improved ABTS radical cation decolorization
assay. Free Rad Biol Med 1999;26:1231–7
Thank you!
Dear Duchuy,
I am sending you the ABTS protocol that I have used in my experiments:
Firstly, the stock solutions of 7 mmol/L ABTS solution and 2.45 mmol/L of potassium persulfate solution were prepared. The working solution was then prepared by mixing 3 mL of each stock solution and allowing them to react for 12-16 h at room temperature (25 °C) in the dark. The solution was then diluted by mixing 4.0-4.5 mL ABTS radical solution with 250 mL distilled water to obtain an absorbance of 0.70 at 734 nm using the spectrophotometer.
Sincerely,
Acácio Zielinski
Hello!
What we do is the following:
- ABTS solution: 192 mg of ABTS in distiled water to 50 mL. Store this solution in ambar flask.
- Potassium persulfate (140 mM): Mix 378.4 mg of the salt with 10 mL of distilled water. Store in ambar flask
ABTS*: The day before the experiment mix in another ambar flask 5 mL of the ABTS solution with 88 uL of the potassium persulfate and leave it at room temperature for 16 hours.
Then you have to dilute 1 mL of the ABTS* to almost 100 mL of methanol. Be careful in this step because you will reach absorbances around 0.750.. then you have to add amounts of methanol to reach the 0.700 +/- 0.050. If you pass below this SD, you can add 10 uL of the ABTS* and measure again.
I hope this can help you, best regards.
Hi, can we use some other compound in place of Potassium persulphate?
Dear Sandeep Rai Kaushik,
I think you must follow the right protocol (using Potassium persulfate), just because you need to compare your result with another result from other journal (they using standard protocol) to make your own conclusion.
Thanks,
Hey everyone, I used distilled water to dilute the working solution and also to prepare stock solutions. It is ok right??
Another quick question, it is ok to prepare Trolox standard stock using 100% methanol right?
Could sodium persalfate be used insead of potassium persulfate?!
and how much sample is used with 1 ml of ABTS*? @Felipe
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