2.5. Off-line spectrophotometric DPPH assay
The antioxidant capacities of sixteen tea samples were evaluated by
off-line DPPH radical scavenging assay. The method is based on the reduction
of the relatively stable radical, DPPH, to the formation of a non
radical form in the presence of hydrogen donating antioxidant. The
tea samples showed antioxidant activity by the reduction of purple
colored DPPH to the yellow colored diphenylpicrylhydrazine derivatives.
DPPH radical scavenging capacity was estimated according to
Brand-Williams, Cuvelier, and Berset (1995) and Shyu and Hwang
(2002) with slight modifications. In the assay, 1 mL of diluted extract
was mixed with 2 mL of 0.1 mmol/L solution of DPPH in methanol.
The mixture was incubated in the dark at room temperature for
30 min, and the absorbance at 517 nm was measured. All tests were
performed in triplicate. The scavenging capacity was calculated as:
(1 − As/Ac) × 100%, where Ac is the absorbance of the control and As
is the absorbance of the tested sample after 30 min. Trolox was used
as standard. Free radical scavenging capacities of tea samples were
expressed as mM Trolox equivalent and IC50 values (concentration of
samples required to scavenge 50% of DPPH radicals).
2.5. Off-line spectrophotometric DPPH assayThe antioxidant capacities of sixteen tea samples were evaluated byoff-line DPPH radical scavenging assay. The method is based on the reductionof the relatively stable radical, DPPH, to the formation of a nonradical form in the presence of hydrogen donating antioxidant. Thetea samples showed antioxidant activity by the reduction of purplecolored DPPH to the yellow colored diphenylpicrylhydrazine derivatives.DPPH radical scavenging capacity was estimated according toBrand-Williams, Cuvelier, and Berset (1995) and Shyu and Hwang(2002) with slight modifications. In the assay, 1 mL of diluted extractwas mixed with 2 mL of 0.1 mmol/L solution of DPPH in methanol.The mixture was incubated in the dark at room temperature for30 min, and the absorbance at 517 nm was measured. All tests wereperformed in triplicate. The scavenging capacity was calculated as:(1 − As/Ac) × 100%, where Ac is the absorbance of the control and Asis the absorbance of the tested sample after 30 min. Trolox was usedas standard. Free radical scavenging capacities of tea samples wereexpressed as mM Trolox equivalent and IC50 values (concentration ofsamples required to scavenge 50% of DPPH radicals).
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