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. 2001 May;54(5):356–361. doi: 10.1136/jcp.54.5.356

Method for the measurement of antioxidant activity in human fluids

D Koracevic 1, G Koracevic 1, V Djordjevic 1, S Andrejevic 1, V Cosic 1
PMCID: PMC1731414  PMID: 11328833

Abstract

Aim—To develop a new, simple, and cheap method for estimating antioxidant activity in human fluids.

Methods—The assay measured the capacity of the biological fluids to inhibit the production of thiobarbituric acid reactive substances (TBARS) from sodium benzoate under the influence of the free oxygen radicals derived from Fenton's reaction. A solution of 1 mmol/litre uric acid was used as standard.

Results—The following mean (SD) antioxidative activities were found (as uric acid) in the various biological fluids: serum, 2.04 (0.20) mmol/litre; urine, 176.5 (25.6) µmol/litre; cerebrospinal fluid, 95.0 (26.9) µmol/litre; aqueous humour oculi, 61.25 (9.9) µmol/litre; saliva, 838.5 (48.2) µmol/litre; tears, 247.0 (17.0) µmol/litre; ascites fluid, 270.0 (63.3) µmol/litre; kidney cyst fluid, 387.1 (28.1) µmol/litre. Small samples of the biological material were needed for the analyses: 10 µl of serum and 50–100 µl of other body fluids. In the sera of 48 healthy individuals there was a significant positive correlation between values obtained with the Randox method (as a reference method) and the new method proposed here (correlation coefficient, 0.8728; mean difference between methods, <0.4%).

Conclusions—This method is easy, rapid, reliable, and practical for the routine measurement of total antioxidant activity in serum and other human body fluids. Small samples of biological material are needed for the analyses and the results are comparable with the reference (Randox) method.

Key Words: antioxidant activity • free oxygen radicals • human fluids

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Figure 1 Influence of the concentration of reagents on antioxidant activity (AOA). Na benzoate was used at 5–40 mM (while Fe–EDTA was kept at 1 mM and H2O2 at 10 mM). H2O2 was used at 5–40 mM (while Fe–EDTA was kept at 1 mM and Na benzoate at 10 mM). Fe–EDTA was used at 0.5–4 mM (while H2O2 was kept at 10 mM and Na benzoate at 10 mM).

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Figure 2 Influence of incubation time on benzoate degradation. AOA, antioxidant activity.

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Figure 3 Frequency distribution of serum antioxidant activity (AOA) in healthy subjects.

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Figure 4 Mean difference between the two methods.

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Figure 5 Mean difference between the Randox method and our proposed method after correction.

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Figure 6 Antioxidant activity (AOA) in human serum.

Selected References

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