A simple, rapid, highly sensitive and reproducible quantification method for plasma malondialdehyde by...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Chemistry

Background:

• Plasma malondialdehyde (MDA) is a marker of lipid peroxidation and oxidative stress.
• Accurate quantification of MDA is crucial for clinical diagnostics and research.
• Existing methods for MDA quantification can be complex or lack sensitivity.

Purpose of the Study:

• To develop a simple, rapid, sensitive, and reproducible method for quantifying plasma MDA.
• To establish a high-performance liquid chromatography (HPLC) assay for MDA detection.

Main Methods:

• The method utilizes the thiobarbituric acid (TBA) reaction followed by reversed-phase HPLC separation.
• Fluorescence detection (excitation/emission: 515/553 nm) is employed for sensitive MDA quantification.
• Plasma samples undergo a simplified preparation involving TBA reaction and heating, with direct injection into the HPLC system.

Main Results:

• The developed method demonstrates high sensitivity with a detection limit of 0.5 fmol per injection.
• Excellent linearity (0.5-500 fmol/injection), precision (within-assay CV 1.2%, between-assay CV 3.1%), and recovery (98.6%) were achieved.
• The method is capable of analyzing 250-300 samples per day with an autosampler.

Conclusions:

• The developed HPLC-based method offers a robust and efficient approach for plasma MDA quantification.
• Its simplicity, sensitivity, and reproducibility make it highly suitable for clinical applications.
• This assay facilitates large-scale analysis of oxidative stress markers in patient samples.