The Measurement Of Benzene-induced Lipid Peroxidation Metabolite Malondialdehyde By High-performance Liquid Chromatography And The Application Of The Crowd

Benzene is the simplest compound in aromatic compounds, which is a good organic solvent widely used in industrial and agricultural production. Chronic exposure to benzene can give rise to damage to the human blood system, mainly for the pancytopenia, anemia, aplastic anemia and leukemia, etc. However, benzene does not cause cancer itself, its carcinogenicity is caused by the toxic metabolites which is the production of metabolic activation in vivo. The toxic metabolites of benzene--hydroquinone and p-benzoquinone can generate reactive oxygen species (ROS) in the bone marrow, which can trigger lipid peroxidation chain reaction to produce hydroxyl peroxide metabolites. The hydroxyl peroxide metabolites can generate some small molecule end metabolites--α,β-unsaturated aldehydes by the non-enzymatic degradation, such as Malonaldehyde (MDA) and 4-hydroxy-2-nonenal (HNE). In this thesis, we focus on the study the method for determination of MDA in plasma and urine samples by HPLC. This dissertation consists of three parts:1. Through the optimization of chromatographic conditions and sample pre-treatment methods, we established a method for the detection of malondialdehyde (MDA) in human plasma samples by High-Performance Liquid Chromatography after derivatisation with 2-thiobarbituric acid (TBA). The retention time and detection limit of TBA-MDA were 4.1min and 0.043nmol/mL, respectively. In this study, we used a lower temperature (80℃) of derivative reaction than before, which overcome the shortcomings of introducing non-sampling inherent MDA due to the high reaction temperature. we found that the method was rapid and had better sensitivity. This method appears simple, accurate and precious.2. We established a method for the detection of malondialdehyde (MDA) in human plasma samples by High-Performance Liquid Chromatography after derivatisation with 2,4-dinitrophenylhydrazine (DNPH), after the optimization of chromatographic conditions and sample pre-treatment methods. The retention time and detection limit of DNPH-MDA were 25.9min and 0.098nmol/mL, respectively. The samples were derivatised with DNPH since this procedure proceeds rapidly under mild acid pH and temperature conditions and the resulting derivatives are unique for a given aldehyde.3. A selective and sensitive method based on derivatisation with 2,4-dinitrophenylhydrazine (DNPH) and consecutive HPLC gradient separation is described for the determination of malondialdehyde (MDA) in urine. The retention time and detection limit of DNPH-MDA were 8.9min and 0.017nmol/mL, respectively. In this study, MDA in urine measured derivatisation with DNPH at 37℃, and the Preparation of urine samples involved a one-step derivatisation/extraction procedure. This method appears simple, rapid and suitable for the determination of a large number of samples.