1. An inter-laboratory validation of methods of lipid peroxidation measurement in UVA-treated human plasma samples.
- Author
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Breusing N, Grune T, Andrisic L, Atalay M, Bartosz G, Biasi F, Borovic S, Bravo L, Casals I, Casillas R, Dinischiotu A, Drzewinska J, Faber H, Fauzi NM, Gajewska A, Gambini J, Gradinaru D, Kokkola T, Lojek A, Luczaj W, Margina D, Mascia C, Mateos R, Meinitzer A, Mitjavila MT, Mrakovcic L, Munteanu MC, Podborska M, Poli G, Sicinska P, Skrzydlewska E, Vina J, Wiswedel I, Zarkovic N, Zelzer S, and Spickett CM
- Subjects
- Chromatography, High Pressure Liquid methods, Chromatography, High Pressure Liquid standards, Chromatography, Liquid methods, Chromatography, Liquid standards, Clinical Chemistry Tests methods, Enzyme-Linked Immunosorbent Assay methods, Enzyme-Linked Immunosorbent Assay standards, Humans, Mass Spectrometry methods, Mass Spectrometry standards, Plasma chemistry, Reproducibility of Results, Sensitivity and Specificity, Ultraviolet Rays, Aldehydes analysis, Clinical Chemistry Tests standards, Isoprostanes analysis, Lipid Peroxidation physiology, Malondialdehyde analysis, Plasma radiation effects
- Abstract
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.
- Published
- 2010
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