Improved analysis of malondialdehyde in human body fluids

Area of Science:

• Biochemistry
• Analytical Chemistry

Background:

• Lipid peroxidation is a marker of oxidative stress.
• The thiobarbituric acid (TBA) assay is widely used to measure lipid peroxidation.
• Artefactual oxidative degradation can occur during the TBA assay.

Purpose of the Study:

• To modify the TBA assay to minimize artefactual lipid peroxidation.
• To investigate the effects of oxygen exclusion and BHT on TBA assay results.
• To adapt the assay for preliminary screening of human body fluids.

Main Methods:

• Modified the TBA assay by depleting oxygen using argon gassing.
• Examined the effect of BHT concentration on TBA-MDA condensation product formation.
• Measured n-butanol extracts using a multititer plate reader.

Main Results:

• Exclusion of oxygen decreased TBARS production in plasma samples.
• High concentrations of BHT also reduced TBARS production.
• Oxygen exclusion had no additional effect at BHT concentrations above 3 mmol/l.

Conclusions:

• The modified TBA assay minimizes artefactual lipid peroxidation.
• The optimized method is suitable for preliminary screening of lipid peroxidation in human body fluids.
• This enhanced assay provides more reliable measurements of oxidative stress markers.