Determination of malondialdehyde in biological samples by solid-phase extraction and high-performance liquid chromatography.
An analytical procedure for determination of malondialdehyde in tissue homogenates and blood serum was developed. A reaction with 2,4-dinitrophenylhydrazine is used followed by cleaning up of the derivative by solid-phase extraction. The samples were analyzed by isocratic high-performance liquid chromatography (HPLC) using a narrow-bore HPLC-column. A good separation of the 1-pyrazole peak from that of 2,4-dinitrophenylhydrazine was observed. A high linear dependence was established by the concentration of 1-pyrazole in the range of 10-5000 ng/ml. The detection limit of the method applied for tissue homogenates and blood serum was approximately 10 ng/ml or lower, and RSD of the method was 9% (n = 8). The peak of 1-pyrazole for these samples was well separated from the other matrix peaks. Experiments carried out evaluated that the solid-phase extraction might be an effective step of the sample preparation, significantly increasing the selectivity of the analysis and the life-time of the column. The method seems to be applicable for determination of malondialdehyde in different biological samples.
Bakalova
R
R
Department of Medical Physics and Biophysics, Medical University, Sofia, Bulgaria. rumi@akita-noken.go.jp
Mileva
M
M
Kotsev
C
C
Bardarov
V
V
Ribarov
S
S
D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008315:Malondialdehyde / Q000032:analysis* / Q000097:blood; D010659:Phenylhydrazines; D051381:Rats; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D007202:Indicators and Reagents; D010659:Phenylhydrazines; C004787:2,4-dinitrophenylhydrazine; D008315:Malondialdehyde