Electrochemical Detection Of 3-Chloro-1,2-Propandiol In Food And Its Toxicity Assessment

3-MCPD belongs to the chloropropanol chemicals which are formed by the reaction between residual vegetable fat and hydrochloric acid in the process of producing HVP. In recent years, HVP as a common savory ingredient is widely added to soy sauce. Therefore,3-MCPD has been detected in a wide range of blended soy sauces. Animal studies show that the main toxity of 3-MCPD is carcinogenicity, reproductive toxicity, nephrotoxicity. In this paper, we establish MIP film-based electrochemical sensor and vivo and vitro experimental evaluation of 3-MCPD toxicity hazards, aiming at building a simple and rapid 3-MCPD detection methods and exploring new biomarkers.1. In this work, a novel electrochemical sensor for 3-MCPD detection based on the Au NPs/GCE coated with a molecular imprinted polymer(MIP) film was constructed.p-Aminothiophenol(p-ATP) and 3-MCPD were self-assembled on a Au NPs/GCE surface,and then a MIP film was formed by electropolymerization. The 3-MCPD template combined with p-ATP through hydrogen bonding during self-assembly and electropolymerization. Then,the cavities matching 3-MCPD were remained after the removal of the template. The MIP sensor was characterized by CV, DPV and SEM. The DPV current was linear with the log of the 3-MCPD concentration in the range from 1.0×10-17 to 1.0×10-13mol/L(R2=0.9939), and the LOD was 3.8×10-18 mol/L(S/N=3). The average recovery rate of 3-MCPD from the spiked soy sauces was between 95.0% and 106.4%(RSD<3.49%). Practically, the sensor showed ideal selectivity, sensitivity, stability and reproducibility.2. 8-OH-d G has been acknowledged as a typical biomarker for oxidative DNA damage.We investigate here the selective cytotoxicity of 3-MCPD on HEK293 cells by analyzing the cell viability, morphological changes, ROS and DNA damage. We further demonstrate that3-MCPD inhibits growth of cells and induces 8-OH-d G generation via ROS-mediated oxidative DNA damage in HEK293 cells. To provide a detection system, we fabricated a modified electrode with poly(3-acetylthiophene)(P3AT), for electrochemical detection of8-OH-d G in damaged cells. Using electropolymerization cyclic voltammetry(CV), we deposited 3-acetylthiophene(3-AT) on a glassy carbon electrode. The conducting polymer,P3 AT, greatly enhances the peak current via dramatic electrocatalytic effect on the chemical behavior of 8-OH-d G. The enrichment potential, time, scan rate, and p H value of supporting electrolyte were optimized. DPV currents maintain a linear relationship with 8-OH-d G concentration between 0.5-35 μmol/L, with a correlative coefficient of 0.9963. We estimate the detection limit of 8-OH-d G to be 31.3 nmol/L(S/N=3). This proposed modified electrode had ideal stability and reproducibility, making it an optimal candidate for detecting 8-OH-d G.We performed detection on real cell samples with satisfactory results.3. During the animal study, we evaluated the effect of 3-MCPD on behavior, body weight,volume diet, tissue, urine and serum biochemical index of rats, and searched for biomarkers in urine through 1H NMR metabolomics. The results revealed that 3-MCPD-dosed rats looked sluggish, ate less and lost significant amounts of weight. The kidneys-body ratio and testes-body ratio for the 3-MCPD-treated group has a significant increase and decrease,respectively. Urinalysis showed significant increase in urinary NAG and GAL activity in3-MCPD-treated rats. BUN and serum creatinine values were increased significantly. Severe focal tubule hydropic degeneration and tubular cell hyperplasia was observed and glomerular morphology was incomplete in high-dosed group. A mild to severe decrease in the spermatozoa and sustentacular cells number in the seminiferous tubules was observed from controls to high-dosed group. Based on the metabolomic as the core, overall metabolic phenotype and related metabolic markers were investigated through chemometrics. 10 kinds of biomarkers and four related metabolic pathways were selected.