Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 832, Issue 1, Pages 103-108Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2005.12.041
Keywords
malonaldehyde; 9-fluorenylmethoxycarbonyl hydrazine; plasma; fluorescence detection
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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 4h at 50 degrees C. The derivatized samples were analyzed by HPLC using a reversed-phase C-18 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. (c) 2006 Elsevier B.V. All rights reserved.
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