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Updated: Apr 17, 2026
Evaluation of Oxidative Stress in Biological Samples Using the Thiobarbituric Acid Reactive Substances Assay
Published on: May 12, 2020
Determination of seminal plasma malondialdehyde by high-performance liquid chromatography in smokers and non-smokers
Objectives:
The aim of this study was to establish and validate a high-performance liquid chromatography method for the determination of malondialdehyde in seminal plasma in smokers and non-smokers and to find possible differences between the two groups.
Background:
Malondialdehyde is used as a diagnostic marker of lipid peroxidation and indicator of oxidative stress. Smoking is suspected to be responsible for an increase in its level. Malondialdehyde has been thought to have cytotoxic and damaging effects.
Methods:
Semen samples were obtained from male partners of couples requesting a fertility evaluation. Malondialdehyde was derivatized with 2-thiobarbituric acid. The malondialdehyde-2-thiobarbituric acid complex was determined by liquid chromatography with fluorescence detection. The mobile phase consisted of 20% ethanol in 25-mmol/L potassium dihydrogenphosphate (v/v), pH 6.00 ± 0.05.
Results:
Analytical performance was satisfactory. Malondialdehyde levels were as follows: 1.50 ± 0.55 µmol/L in all patients, 1.40 ± 0.57 µmol/L in smokers, and 1.50 ± 0.53 µmol/L in non-smokers.
Conclusion:
The method presented here is sensitive and accurate for seminal plasma malondialdehyde determination. Our results showed a relationship between sperm motility and the malondialdehyde level in all patients and non-smokers. Malondialdehyde may induce poor sperm functionality and negatively affect the fertilization processes (Tab. 1, Fig. 1, Ref. 23).
