Quantifying Leaf Antioxidant Activity

For antioxidant activity (TAA), leaf powder (∼100 mg) was homogenized with methanol and incubated for 30 min at 40°C (Re et al., 1999 (link)). After centrifugation (15 000g, 15 min at 4°C), the supernatant was mixed with an ABTS (2,20-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) solution. The absorbance was read at 734 nm using a Jenway 7305 spectrophotometer. Antioxidant activity was determined using a gallic acid standard curve (y=0.0002x + 0.173, R² = 0.984).

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Castro H., Dias M.C., Castro M., Loureiro J, & Castro S. (2023). Impact of genome duplications in drought tolerance and distribution of the diploid-tetraploid Jasione maritima. Frontiers in Plant Science, 14, 1144678.

Publication 2023

Abts Antioxidant activity Centrifugation Gallic acid Leaf Methanol Powder Sulphonic acid

Corresponding Organization : University of Coimbra

Top 5 similar protocols

Variable analysis

independent variables

Leaf powder (∼100 mg)

dependent variables

Antioxidant activity (TAA)

control variables

Methanol incubation time (30 min)
Methanol incubation temperature (40°C)
Centrifugation speed (15,000g)
Centrifugation time (15 min)
Centrifugation temperature (4°C)
Spectrophotometer (Jenway 7305)

positive control

Gallic acid standard curve (y=0.0002x + 0.173, R^2 = 0.984)

negative control

Not explicitly mentioned

Annotations

Based on most similar protocols

Optimize homogenization by using a vortex for 1 min to ensure complete extraction of antioxidants (Protocol 1, Protocol 2).

Incubate the samples in a shaker incubator at 30°C for 1 hour to maximize antioxidant extraction (Protocol 1, Protocol 2).

Centrifuge the samples at 14,560 x g for 15 min at 5°C to ensure complete separation of the supernatant (Protocol 1, Protocol 2).

Use a spectrophotometer to measure the absorbance of the samples at 734 nm, which is the optimal wavelength for the ABTS assay (Input protocol, Protocol 1, Protocol 2).

Prepare a gallic acid standard curve to quantify the antioxidant activity in terms of gallic acid equivalents (Input protocol, Protocol 4).

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