Rapid quantification of malondialdehyde in plasma by high performance liquid chromatography–visible detection
Abstract
Background Malondialdehyde (MDA) is one of the better-known secondary products of lipid peroxidation, and it is widely used as an indicator of cellular injury. The employment of the thiobarbituric acid reactive substances (TBARS) technique to measure MDA has received criticism over the years because of its lack of specificity. Thus, a specific and reliable method for MDA determination in plasma by high performance liquid chromatographic (HPLC)-VIS was validated; alkaline hydrolysis, n-butanol extraction steps and MDA stability were established. Methods The plasma underwent alkaline hydrolysis, acid deproteinization, derivatization with TBA and n-butanol extraction. After this, MDA was determined at 532 nm by HPLC-VIS. The method was applied to 65-year-old subjects from a retirement home. Results The assay was linear from 0.28 to 6.6 microM. The reproducibility of intra-run was obtained with CV% Conclusion The method is reproducible, accurate, stable, sensitive, and can be used in the routines in clinical laboratories. Besides, this technique presents advantages such as the complete release of protein bound MDA with the alkaline hydrolysis step, the removal of interferents with n-butanol extraction, mobile phase without phosphate buffer and rapid analytical processes and run times.
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Further methods such as HPLC with UV or fluorescence detection have been developed to improve specificity of MDA measurement.
“Beside this assay, further methods have been developed in order to improve the specificity of MDA measurement, such as HPLC combined with UV or fluorescence detection [182,184].”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [182]
The TBARS assay is unspecific and measures total oxidative stress rather than MDA alone.
“This approach is the method of choice for the quantification of free MDA due to simplicity and low costs, but is often criticized as being unspecific and rather measuring total oxidative stress instead of MDA alone [181,182].”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [182]
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Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases
Kehm, R., Baldensperger, T., Raupbach, J., & Höhn, A. (2021) · Redox Biology · doi:10.1016/j.redox.2021.101901 · Cited by 348 · see citing works
“This approach is the method of choice for the quantification of free MDA due to simplicity and low costs, but is often criticized as being unspecific and rather measuring total oxidative stress instead of MDA alone [181,182].”
MethodSafavi, A., Ahmadi, R., & Ramezani, A. M. (2018) · Microchemical Journal · doi:10.1016/j.microc.2018.07.036 · Cited by 101 · see citing works
- Authors
- Grotto, D., Santa Maria, L. D., Boeira, S., Valentini, J., Charão, M. F., Moro, A. M., Nascimento, P. C., Pomblum, V. J., & Garcia, S. C.
- Affiliations
- Universidade Federal de Santa Maria (BR)
- Venue
- Journal of Pharmaceutical and Biomedical Analysis
- Publisher
- Elsevier BV
- Published
- 2007-01
- Volume
- 43
- Issue
- 2
- Pages
- 619-624
- ISSN
- 0731-7085 (ISSN-L)
- OpenAlex
- W2084727563
- Cited by
- 351
- References
- 0
- Citations by year
- 2026: 12 · 2025: 18 · 2024: 26 · 2023: 13 · 2022: 12 · 2021: 17 · 2020: 24 · 2019: 26 · 2018: 21 · 2017: 20 · 2016: 32 · 2015: 22 · 2014: 16 · 2013: 20 · 2012: 21
- FWCI
- 4.73
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Pharmaceutical and Biomedical Analysis (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutscher Akademischer Austauschdienst
- Topics
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- malondialdehyde, plasma, lipid peroxidation, alkaline hydrolysis, n-butanol extraction, method validation, sample stability, older adults, oxidative stress biomarkers
- MeSH terms
- Aged, Brazil, Chromatography, High Pressure Liquid, Female, Humans, Hydrolysis, Male, Malondialdehyde, Reference Values, Sensitivity and Specificity, Sodium Hydroxide, Solvents, Spectrophotometry, Thiobarbiturates, Time Factors, Reproducibility of Results, Linear Models, 1-Butanol
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