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Anal Biochem ; 441(2): 109-14, 2013 Oct 15.
Article in English | MEDLINE | ID: mdl-23872008

ABSTRACT

The use of potentiometry to measure plasma antioxidant capacity to contribute to oxidative stress evaluation is presented. In this assay, plasma (n=60) diluted (0.3 to 1 ml) in phosphate buffer, pH 7.4, NaCl 9%, was submitted to potentiometry. A platinum wire was the working electrode and saturated calomel the reference. The results are presented as the difference between sample and buffer potential (ΔE). ΔE presented a good inverse correlation with added increasing concentrations of ascorbate (2.5-75 µmol/L; R=-0.99), urate (9.0-150 µmol/L; R=-0.99), and bilirubin (0.78-13 µmol/L; R=-0.99). Increase in the antioxidant capacity decreased ΔE. Depletion of the antioxidant capacity by tert-butylhydroperoxide (6.5-50 µmol/L) presented a direct correlation (0.97) with ΔE. Furthermore, ΔE presented an inverse correlation (R=-0.99) with increased antioxidant capacity of plasma (FRAP) induced by the addition of ascorbate (2.5-75 µmol/L). The response of the potentiometric method proved be adequate for measuring the plasma antioxidant depletion induced by acute exhaustive exercise in rats (control, n=15; exercised, n=15). This exercise decreased the concentration of urate (p<0.05), decreased FRAP (p<0.5), increased TBARS (p<0.5), and decreased the potentiometer sensor response (p=6.5×10⁻³). These results demonstrate the adequacy of potentiometry for evaluating the antioxidant capacity of blood plasma samples.


Subject(s)
Antioxidants/metabolism , Plasma/metabolism , Potentiometry/methods , Adolescent , Animals , Antioxidants/analysis , Ascorbic Acid/blood , Ascorbic Acid/metabolism , Exercise , Humans , Male , Oxidative Stress , Rats , Rats, Wistar , Thiobarbituric Acid Reactive Substances/analysis , Thiobarbituric Acid Reactive Substances/metabolism , Uric Acid/blood , Uric Acid/metabolism , Young Adult
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