The previously described method was followed to measure hydrogen peroxide content (H2O2) in plant extracts [145 (link)]. The freeze-dried samples (0.3 g each) were blended with 5 mL 0.1% trichloroacetic acid (TCA) in an ice bath and centrifuged at 12,000 rpm for 20 min. The obtained supernatant (0.5 mL) was combined with 10 mM potassium phosphate buffer (0.5 mL, pH 7.0) and 1 M potassium iodide (1 mL). The reaction mixture intensity was recorded at 390 nm using a spectrophotometer (MultiskanTM GO Microplate Spectrophotometer, Thermo Fisher Scientific, Waltham, MA, USA).
The lipid peroxidation level of freeze-dried samples (0.2 g) was determined by applying the thiobarbituric acid (TBA) test [146 (link)]. The mixture contained 0.5 mL of 0.1% TCA extract that was added to 1 mL of 0.5% TBA (prepared in 20% TCA). The mixture was incubated in boiling water (95 °C, 30 min), followed by cooling in an ice bath (10 min). Subsequently, the homogenate was centrifuged (12,000 rpm, 5 min) and the supernatant absorbance was measured at 532 and 600 nm. The lipid peroxidation level was calculated according to a standard curve (MultiskanTM GO Microplate Spectrophotometer, Thermo Fisher Scientific, Waltham, MA, USA).
The lipid peroxidation level of freeze-dried samples (0.2 g) was determined by applying the thiobarbituric acid (TBA) test [146 (link)]. The mixture contained 0.5 mL of 0.1% TCA extract that was added to 1 mL of 0.5% TBA (prepared in 20% TCA). The mixture was incubated in boiling water (95 °C, 30 min), followed by cooling in an ice bath (10 min). Subsequently, the homogenate was centrifuged (12,000 rpm, 5 min) and the supernatant absorbance was measured at 532 and 600 nm. The lipid peroxidation level was calculated according to a standard curve (MultiskanTM GO Microplate Spectrophotometer, Thermo Fisher Scientific, Waltham, MA, USA).
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