An inter-laboratory validation of methods of lipid peroxidation measurement in UVA-treated human plasma samples

Free Radic Res. 2010 Oct;44(10):1203-15. doi: 10.3109/10715762.2010.499907.


Lipid peroxidation products like malondialdehyde, 4-hydroxynonenal and F(2)-isoprostanes are widely used as markers of oxidative stress in vitro and in vivo. This study reports the results of a multi-laboratory validation study by COST Action B35 to assess inter-laboratory and intra-laboratory variation in the measurement of lipid peroxidation. Human plasma samples were exposed to UVA irradiation at different doses (0, 15 J, 20 J), encoded and shipped to 15 laboratories, where analyses of malondialdehyde, 4-hydroxynonenal and isoprostanes were conducted. The results demonstrate a low within-day-variation and a good correlation of results observed on two different days. However, high coefficients of variation were observed between the laboratories. Malondialdehyde determined by HPLC was found to be the most sensitive and reproducible lipid peroxidation product in plasma upon UVA treatment. It is concluded that measurement of malondialdehyde by HPLC has good analytical validity for inter-laboratory studies on lipid peroxidation in human EDTA-plasma samples, although it is acknowledged that this may not translate to biological validity.

Publication types

  • Multicenter Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Aldehydes / analysis*
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, High Pressure Liquid / standards
  • Chromatography, Liquid / methods
  • Chromatography, Liquid / standards
  • Clinical Chemistry Tests / methods
  • Clinical Chemistry Tests / standards*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / standards
  • Humans
  • Isoprostanes / analysis*
  • Lipid Peroxidation / physiology*
  • Malondialdehyde / analysis*
  • Mass Spectrometry / methods
  • Mass Spectrometry / standards
  • Plasma / chemistry
  • Plasma / radiation effects*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Ultraviolet Rays


  • Aldehydes
  • Isoprostanes
  • Malondialdehyde
  • 4-hydroxy-2-nonenal