I am preparing DPPH solution for antioxidant assay. As the reagent is costly n sensitive enough to prepare it freshly, so may i know the exact way with detailed step wise protocol to prepare the reagent .....
Will be greatly thankful for getting the answer
Dear Anup,
We use this verified protocol:
0.1 mL of a water solution of the antioxidant tested (1 mM) was added
to 2.9 mL of an ethanolic solution of DPPH (62.1 μM) to give the
final concentrations 33.3 and 60 μM for the antioxidant and DPPH,
respectively.
Dear Anup,
You can prepare the solution with making 0.1 N DPPH in methanol.
Best regards,
You can use 0.1mM DPPH in MeOH.
But make sure your drugs or extract is colourless.
Good Luck
Dissolve 1.9 mg DPPH in 100 mL MeOH and shake for few mints to ensure that all DPPH dissolved in solvent.
Dear Anup,
we dissolve 0.4mg DPPH in 1 ml absolute ethanol. keep it away from light and ensure that all DPPH dissolved in solvent.
Best regards
The usual 0.004% dpph solution is an equivalent of 100mM. Hence % solutions such as this give a molar concentration since the molar mass of dpph is known.
In addition, and in my personal opinion and experience which is supported by studies such as Mishra et al and others, there's still the need for standardisation of dpph concentrations (and actual assay mix ratios) used in various studies to ensure uniformity in antiradical capacity reports on extracts, compounds etc.
