New method for HPLC separation and fluorescence detection of malonaldehyde in normal human plasma.
A new method for the detection of free and total malonaldehyde (MDA) in human plasma samples based on the derivatization of MDA with 9-fluorenylmethoxycarbonyl hydrazine (FMOC-hydrazine) in an acidic medium was developed. Derivatization was achieved after 4 h at 50 degrees C. The derivatized samples were analyzed by HPLC using a reversed-phase C18 column with fluorescence detection (Ex=270 nm, Em=310 nm). The benefit of this direct injection of deproteinized plasma is to avoid the use of an internal standard. The detection limit was 0.1 pmol (4.0 nmol/L). The recovery of MDA spiked in different human plasma samples was 95.3% (n=25; R.S.D. 5.1%) for the hydrolysation procedure. The total and free MDA in plasma of 15 healthy male volunteers are 426+/-29.8 nmol/L and 153+/-9.6 nmol/L, respectively.
Mao
Jianjun
J
Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, PR China.
Zhang
Hongwu
H
Luo
Jia
J
Li
Li
L
Zhao
Rui
R
Zhang
Renen
R
Liu
Guoquan
G
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008297:Male; D008315:Malondialdehyde / Q000097:blood*; D012015:Reference Standards; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D008315:Malondialdehyde