On-line antioxidant activity determination: comparison of hydrophilic and lipophilic antioxidant activity using the ABTS*+ assay.
The ABTS/H(2)O(2)/HRP decoloration method is capable of determining both hydrophilic (in buffered media) and lipophilic (in organic media) antioxidant properties in complex samples. Now, we have adapted this method for on-line chromatographic determination. The easy, rapid and controlled generation of the ABTS radical and its great stability in buffered and organic media were important characteristics in the measurement of antioxidant activities. The HPLC-ABTS method used two pumps (one for isocratic eluting-phase and the other for preformed ABTS radical) and an UV-VIS diode array detector. The dual analysis of samples -- conventional (with UV-VIS detection) and ABTS-scavenging (at 600 nm) -- provided valuable on-line information about the correspondence between the presence of a determined compound and its possible antioxidant activity, and was applicable to both hydrophilic and lipophilic antioxidants (HAA and LAA). A comparison between HAA and LAA determined by the end-point method and by the on-line HPLC method is presented. The application to juices showed that both methods are suitable, sensitive and selective, gave similar values, and the HPLC-ABTS method contributed additional information about the antioxidant activity profile.
Department of Plant Biology (Plant Physiology), Campus Espinardo, University of Murcia, 30100-Murcia, Spain.
Antonio
A
Cano
Olga
O
Alcaraz
Manuel
M
Acosta
Marino B
MB
Arnao
D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology; D052160:Benzothiazoles; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005609:Free Radicals / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007202:Indicators and Reagents; D009862:Online Systems; D013053:Spectrophotometry; D013329:Structure-Activity Relationship; D013451:Sulfonic Acids
D000975:Antioxidants; D052160:Benzothiazoles; D005609:Free Radicals; D007202:Indicators and Reagents; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D001205:Ascorbic Acid