An inter-laboratory validation of methods of lipid peroxidation measurement in UVA-treated human plasma samples.
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.
Breusing
Nicolle
N
Institute of Biological Chemistry and Nutrition, University of Hohenheim, Stuttgart, Germany.
Grune
Tilman
T
Andrisic
Luka
L
Atalay
Mustafa
M
Bartosz
Grzegorz
G
Biasi
Fiorella
F
Borovic
Suzana
S
Bravo
Laura
L
Casals
Isidre
I
Casillas
Rosario
R
Dinischiotu
Anca
A
Drzewinska
Joanna
J
Faber
Heidemarie
H
Fauzi
Norsyahida Mohd
NM
Gajewska
Agnieszka
A
Gambini
Juan
J
Gradinaru
Daniela
D
Kokkola
Tarja
T
Lojek
Antonin
A
Luczaj
Wojciech
W
Margina
Denisa
D
Mascia
Cinzia
C
Mateos
Raquel
R
Meinitzer
Andreas
A
Mitjavila
María Teresa
MT
Mrakovcic
Lidija
L
Munteanu
Maria Cristina
MC
Podborska
Martina
M
Poli
Giuseppe
G
Sicinska
Paulina
P
Skrzydlewska
Elzbieta
E
Vina
Jose
J
Wiswedel
Ingrid
I
Zarkovic
Neven
N
Zelzer
Sieglinde
S
Spickett
Corinne M
CM
D000447:Aldehydes / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D019963:Clinical Chemistry Tests / Q000379:methods / Q000592:standards*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D006801:Humans; D028421:Isoprostanes / Q000032:analysis*; D015227:Lipid Peroxidation / Q000502:physiology*; D008315:Malondialdehyde / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards; D010949:Plasma / Q000737:chemistry / Q000528:radiation effects*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014466:Ultraviolet Rays
D000447:Aldehydes; D028421:Isoprostanes; D008315:Malondialdehyde; C027576:4-hydroxy-2-nonenal