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Quantification of Antioxidant Ability Against Lipid Peroxidation with an ‘Area Under Curve’ Approach

  • Original Paper
  • Published:
Journal of the American Oil Chemists' Society

Abstract

When oxygen is passed through a linoleic acid (LA) emulsion containing copper(II), primary (hydroperoxides) and secondary oxidation products (aldehydes and ketones) are formed, monitored by ferric thiocyanate (Fe(III)-SCN) and thiobarbituric acid reactive substances (TBARS) colorimetry, respectively. As total antioxidant capacity (TAC) against lipid peroxidation was not quantified before, both methods were adapted to an ‘area under curve (AUC)’ approach. LA peroxidation followed pseudo-first order kinetics in aerated emulsions. Absorbance changes as a function of incubation time exhibited sigmoidal curves, enabling the calculation of ‘area under curve’ (AUC) and net AUC = AUCblank−AUCsample, standard calibration curve as net AUC versus concentration, and trolox-equivalent antioxidant capacity of the tested compounds. Garlic extract showed an antioxidative effect on hydroperoxide formation, but a prooxidative effect on TBARS. Although inhibition of lipid peroxidation was described qualitatively before, it was not evaluated quantitatively, e.g., the trolox-equivalent antioxidant capacities (TEAC values) of antioxidants with respect to their inhibitive effect against lipid peroxidation were not calculated and compared. Additionally, real-time monitoring of lipid oxidation products requires highly sophisticated but costly instrumental techniques, but no single oxidation product is a direct measure of lipid oxidation or its antioxidative prevention. The AUC approach is the first quantitative method measuring antioxidant protection against lipid oxidation, with a slightly different order of antioxidative effectiveness from reductive assays because of interfacial effects.

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Abbreviations

LA:

Linoleic acid

Fe(III)-SCN:

Ferric thiocyanate

TBA:

Thiobarbituric acid

TBARS:

Thiobarbituric acid-reactive substances

MDA:

Malondialdehyde

AUC:

Area under curve

TEAC:

Trolox-equivalent antioxidant capacity

k 1 :

Pseudo-first order rate constant with respect to hydroperoxides formation (measured by Fe(III)-SCN method in the presence of antioxidant)

k ′1 :

Pseudo-first order rate constant with respect to aldehydes and ketones formation (measured by TBARS method in the presence of antioxidant)

k 0 :

Pseudo-first order rate constant of the control reaction (in the absence of antioxidant)

I,%:

Percentage inhibition

A :

Absorbance, A 500 nm or A 532 nm, proportional to the total concentration of hydroperoxides or secondary products (aldehydes and ketones)

t :

Incubation time

TocH:

Tocopherol

TR:

Trolox

AP:

Ascorbyl palmitate

HQ:

Hydroquinone

TBHQ:

tert-Butyl hydroquinone

TBQ:

tert-Butyl quinone

QR:

Quercetin

MR:

Morin

CT:

Catechin

AA:

Ascorbic acid

FOX:

Ferrous oxidation xylenol orange

PV:

Peroxide value

HAT:

Hydrogen atom transfer

RG:

Raw garlic

ESR:

Electron spin resonance

CIS-MS:

Coordination ion spray-mass spectrometry

ESI-MALDI-TOF:

Electrospray ionization-matrix-assisted laser desorption/ionization-time of flight

TAC:

Total antioxidant capacity

ROS:

Reactive oxygen species

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Acknowledgements

This work was supported by Yildiz Technical University, Research Fund, YTÜ-BAPK Project No: 29-01-02-03. One of the authors (Dr. Apak) would like to thank Istanbul University–Application & Research Center for the Measurement of Food Antioxidants (Istanbul Universitesi Gida Antioksidanlari Olcumu Uygulama ve Arastirma Merkezi) for the development of antioxidant/prooxidant test methods.

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Authors and Affiliations

  1. Department of Chemistry, Faculty of Science and Letters, Kastamonu University, Kuzeykent, 37150, Kastamonu, Turkey

    Temelkan Bakır

  2. Department of Chemistry, Faculty of Science and Letters, Yildiz Technical University, Esenler, 34010, Istanbul, Turkey

    İnci Sönmezoglu

  3. Division of Analytical Chemistry, Department of Chemistry, Faculty of Engineering, Istanbul University, Avcilar, 34320, Istanbul, Turkey

    Reşat Apak

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  1. Temelkan Bakır
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  2. İnci Sönmezoglu
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Correspondence to Reşat Apak.

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Bakır, T., Sönmezoglu, İ. & Apak, R. Quantification of Antioxidant Ability Against Lipid Peroxidation with an ‘Area Under Curve’ Approach. J Am Oil Chem Soc 94, 77–88 (2017). https://doi.org/10.1007/s11746-016-2918-2

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