212
T. Casagrande do Nascimento et al.
Table 9.1
Characteristics of the most relevant in vitro chemical antioxidant capacity assays
Assay
Mechanism
Reaction medium
Radical generator/Reactive
specie/Probe
T (ºC)
Monitoring
(λ)
Advantages
Limitation
References
DPPH
SET (main) and
HAT (marginal
reaction path)
Methanol
none/DPPH/DPPH
25 °C
515 nm
Rapid; Easy to
execute; Evaluation
of hydrophilic and
lipophilic
compounds
Non-competitive;
Radical nonexistent in
the organism; Reacts
different from
physiological ROS and
RNS; Spectral
interference with natural
pigments that absorbs at
the same monitoring
wavelength
Brandt-Williams
et al. (1995)
ABTS
SET
Ethanol or
Phosphate buffer
(pH ~ 7.4)
ABTS
+ Potassium
persulphate/ABTS •+
/ABTS •+
30 °C
750 or 734 nm Rapid; Easy to
execute; Evaluation
of hydrophilic and
lipophilic
compounds; Low
spectral interference
Non-competitive;
Radical nonexistent in
the organism; Reacts
different from
physiological ROS and
RNS
Re et al. (1999)
FRAP
SET
Acetate buffer (pH
~ 3.4)
None (metal reduction)
37 °C
593 nm
Rapid; Easy to
execute
Non-competitive; No
reactive specie; pH
outside physiological
conditions (acid);
Nonspecific reaction
Bezie and Strain
(1996)
RC
SET
Aqueous solution
(pH 11.0)
None (metal reduction)
25 °C
750 nm
Easy to execute;
Many results
available
Non-competitive; No
reactive specie; pH
outside physiological
conditions (basic);
Several compounds
react with
Folin-Ciocalteu reagent
Singlenton and
Joseph (1965)
(continued)