The antioxidant capacity of human plasma was determined by following the oxidation kinetics of the lipid-soluble fluorescent marker BODIPY using 2,20-azobis(4-methoxy-2,4-dimethylvaleronitrile) (MeO-AMVN) as the lipophilic radical initiator. The results are expressed as a total antioxidant performance (TAP) value based on the inhibition of BODIPY oxidation, as determined by the appearance of green fluorescence, with respect to a control sample (phosphatidylcholine with or without delipidized human serum). The suitability of the assay was evaluated on the basis of its precision, reproducibility, and specificity. The intra- and interassay coefficients of variation both were less than 5%. The addition of a representative substrate of plasma peroxidation, phosphatidylcholine, up to 750 lg/ml did not induce significant changes in the TAP value. Also, BODIPY photooxidation was not observed during the experimental time course (220 min). The TAP values of 6 plasma samples from healthy donors were measured and correlated with the main plasma waterand lipid-soluble antioxidants (uric acid and ascorbic acid, a-tocopherol, and carotenoids) and lipid profiles. Significant correlations were found between TAP and uric acid (R = 0.97, P < 0.05) and cholesterol-adjusted a-tocopherol (R = 0.93, P < 0.01). The results confirm that the TAP assay is suitable to measure the antioxidant activity of plasma antioxidants localized in both the lipophilic and hydrophilic compartments
Total antioxidant performance: a validated fluorescence assay for the measurement of plasma oxidizability / G. Beretta, G. Aldini, R. Maffei Facino, R. M. Russell, N. I. Krinsky, K. J. Yeum. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - 354:2(2006), pp. 290-298. [10.1016/j.ab.2006.04.001]
Total antioxidant performance: a validated fluorescence assay for the measurement of plasma oxidizability
G. BerettaPrimo
;G. AldiniSecondo
;R. Maffei Facino;
2006
Abstract
The antioxidant capacity of human plasma was determined by following the oxidation kinetics of the lipid-soluble fluorescent marker BODIPY using 2,20-azobis(4-methoxy-2,4-dimethylvaleronitrile) (MeO-AMVN) as the lipophilic radical initiator. The results are expressed as a total antioxidant performance (TAP) value based on the inhibition of BODIPY oxidation, as determined by the appearance of green fluorescence, with respect to a control sample (phosphatidylcholine with or without delipidized human serum). The suitability of the assay was evaluated on the basis of its precision, reproducibility, and specificity. The intra- and interassay coefficients of variation both were less than 5%. The addition of a representative substrate of plasma peroxidation, phosphatidylcholine, up to 750 lg/ml did not induce significant changes in the TAP value. Also, BODIPY photooxidation was not observed during the experimental time course (220 min). The TAP values of 6 plasma samples from healthy donors were measured and correlated with the main plasma waterand lipid-soluble antioxidants (uric acid and ascorbic acid, a-tocopherol, and carotenoids) and lipid profiles. Significant correlations were found between TAP and uric acid (R = 0.97, P < 0.05) and cholesterol-adjusted a-tocopherol (R = 0.93, P < 0.01). The results confirm that the TAP assay is suitable to measure the antioxidant activity of plasma antioxidants localized in both the lipophilic and hydrophilic compartmentsPubblicazioni consigliate
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