Abstract
An improved system for measuring antioxidant activity via thiobarbituric acid reactive substances and ferric thiocyanate assays is reported, on the basis of oxidation of a linoleic acid (LA) emulsion. Oxidation times were reduced from 20 h to 5 h by increasing the reaction temperature from 37 °C to 50 °C and with an acceptable precision of <10% coefficient of variation (CV). Antioxidants varying in polarity and chemical class—250 µM Trolox, quercetin, ascorbic acid and gallic acid—were used for method optimisation. Further reductions in reaction time were investigated through the addition of catalysts, oxygen initiators or increasing temperature to 60 °C; however, antioxidant activity varied from that established at 37 °C and 20 h reaction time—the method validation conditions. Further validation of the method was achieved with catechin, epicatechin, caffeic acid and α-tocopherol, with results at 50 °C and 5 h comparable to those at 37 °C and 20 h. The improved assay has the potential to rapidly screen antioxidants of various polarities, thus making it useful in studies where large numbers of plant extracts require testing. Furthermore, as this assay involves protection of a lipid, the assay is likely to provide complementary information to well-established tests, such as the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay.
Highlights
Lipid oxidation is problematic in the food industry as it leads to rancidity [1], and is more broadly detrimental to human health as it is implicated in such diseases as atherosclerosis, cancer and arthritis [2]
The latter assay is commonly used for dairy products such as milk or fats [3,4] while the thiobarbituric acid reactive substances (TBARS) assay is used for meat products [5] and in human physiology [6]
Oxidation of a lipid substrate is related to the size of the droplets in an emulsion [25] and to the polydispersity of droplet size
Summary
Lipid oxidation is problematic in the food industry as it leads to rancidity [1], and is more broadly detrimental to human health as it is implicated in such diseases as atherosclerosis, cancer and arthritis [2]. Polyunsaturated fatty acids (PUFAs), such as linoleic acid (LA), are very susceptible to oxidation in the presence of heat, light or trace amounts of metal ions. This oxidation produces harmful primary oxidation products such as peroxides, which breakdown to numerous toxic secondary oxidation products, such as aldehydes, carbonyl compounds, conjugated dienes and furans. Peroxides may be measured by the ferric thiocyanate (FTC) assay The latter assay is commonly used for dairy products such as milk or fats [3,4] while the TBARS assay is used for meat products [5] and in human physiology [6]
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