Determination of malondialdehyde in biological fluids by high-performance liquid chromatography using rhodamine B hydrazide as the derivatization reagent |
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| Affiliation: | 1. School of Materials Science & Engineering, Key Laboratory of Advanced Technologies of Materials (Ministry of Education), Southwest Jiaotong University, Chengdu, 610031, China;2. Analytical and Testing Center, Southwest Jiaotong University, Chengdu, 610031, China;1. Department of Animal Science and Biotechnology, Chungnam National University, Daejeon 34134, South Korea;2. Department of Animal Science and Technology, Sunchon National University, Suncheon 57922, South Korea;3. Department of Agricultural Biotechnology, Center for Food and Bioconvergence, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, South Korea;1. Xinxiang Key Laboratory of Forensic Science Evidence, School of Forensic Medicine, Xinxiang Medical University, Jinsui Road No. 601, Xinxiang 453003, Henan Province, PR China;2. Henan Key Laboratory of Biological Psychiatry, The Second Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, PR China;3. School of Materials Science and Engineering, Henan Normal University, Xinxiang 453003, Henan Province, PR China;4. School of Basic Medicine, Xinxiang Medical University, Jinsui Road No. 601, Xinxiang 453003, Henan Province, PR China |
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| Abstract: | Malondialdehyde (MDA) is a biomarker for lipid peroxidation, and studies of sensitive and selective analytical methods for it are very important for pathological research. The aim of this work was to develop and validate a novel HPLC method for the quantification of MDA in biological fluids using rhodamine B hydrazide (RBH) as the derivatization reagent. After pretreatment and derivatization in acid medium at 50 °C for 40 min, the RBH-derivatized MDA was separated on a Kromasil C18 column at 25 °C and detected by a fluorescence detector at excitation wavelength of 560 nm and emission wavelength of 580 nm. The results showed linearity in the range of 0.8–1500.0 nM with a detection limit of 0.25 nM (S/N = 3). The recovery of MDA from plasma and urine was 91.50 to 99.20%, with a relative standard deviation range of 1.45 to 3.26%. In comparison to other methods reported for the determination of MDA, the proposed method showed superiority in simplicity, more sensitivity, shorter derivatization time, and less interference. The developed method was applied to quantification of MDA in human biological fluids collected from five volunteers with a concentration range of 24.62–245.00 nM. |
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| Keywords: | Determination High-performance liquid chromatography Malondialdehyde Precolumn derivatization Rhodamine B hydrazide Free radicals |
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