Detection Of Fumonision B₁ And Combined Effect With Sterigmatocystin On Hepatic Cell Of Mouse

Part one A novel capillary electrophoresis-chemiluminescence method for determination of fumonisin B₁Objective: Established a series method of using immunoaffinity column purification and enrichment of fumonisin B₁ and using capillary electrophoresis chemiluminescence detection of fumonisin B₁ in maize, and the method were applied to detect fumonisin B₁ in corn.Methods: 1 The fumonisin B₁ standard stock solution diluted with double distilled water to a series of standard solution with the concentration of 100.0μg/ml, 50.0μg/ml,25.0μg/ml,10.0μg/ml and 1.0μg/ml, to detect conditions for selection and optimization. 2 Using acetonitrile: methanol: water（1:1:2） extracted FB₁ from corn samples. Using immune affinity column purification of fumonisin B₁, with water as mobile phase, detected the content of FB₁ with capillary electrophoresis chemiluminescence in corns.Results: 1 Based on the inhibitory effect of fumonisin B₁ on luminol trivalent silver chemiluminescence system, combined with capillary electrophoresis separation technology, to optimize the detection method and the emission conditions: 5.0×10-3mol/L borax buffer solution, the concentration of luminol to 2.0×10-3mol / L, the concentration of Ag（III） for 3.0×10-5mol/L dissolved in 0.01mol/L sodium hydroxide; 2 Under the optimum detection conditions, the linear range of fumonisin B₁ was 1.0μg/ml²00.0μg/ml and the detection limit（S/N=3） was 1.0μg/ml. After 8 times parallel determination of 200.0μg/ml of fumonisin B₁,the peak time and peak height relative standard deviation（n = 8） were 2.64% and 7.86%.The average recovery rate was 86.37% and the coefficient of variation was 7.43%.Conclusions: The experimental application of capillary electrophoresis with chemiluminescence method to detect the fumonisin B₁ in maize had simple operation, rapid detection, and accurate and reliable result. Fumonisin B₁ was detected for this application in corn with satisfactory results.Part two Toxicity and oxidative damage of Fumonisin B₁ and sterigmatocystin on cellObjective: The inquiry fumonisin element and combined processing of sterigmatocystin（ST） on rat hepatocytes injury types, and application of superoxide dismutase（SOD）, malondialdehyde（MDA）, glutathione peroxidase（GSH-Px） index of intracellular protein oxidation injury level in verification.Methods: 1 Determined fumonisin and sterigmatocystin combined exposure dose using CCK-8 cell survival rate in experiment, and the results of five-mixed toxin concentration were applied in following tests. 2 Using the application of q value calculated the combined effect of the correlation coefficient（q）, and determined the type of interaction both toxins. 3 Explained and analysed the correlation between cytotoxicity and oxidative damage of two kinds of toxin. 4 The results of each step of the experiment were verified by the 3 independent repeated experiments.Results: 1 FB₁ can make the cell survival rate decreased significantly with the increase of the concentration, and there is dose dependent（correlation coefficient r=0.990, P=0.001）. After 48 h of ST treatment the cell viability was decreased, and the toxic effects were significantly enhanced with the concentration increasing, there is dose dependence（correlation coefficient r= 0.989, P=0.001）. Both of the two toxins and the combined effects of the two toxins significantly reduced the cell viability, and showed a dose-dependent; 2 After 48 hours of the combined effects of sterigmatocystin and fumonisin, calculated the joint coefficient for q1=1.77, q2=2.01, q3=2.81, q4=2.10. So,when ST concentration was 10μmol/L there was a synergistic effect, when the ST concentration increased to 20.0μmol/L, 40.0μmol / L, 80.0μmol/L there was a significantly enhanced; 3 Cytotoxicity and oxidative damage of the cells showed a significant correlation with the combined effects of the two toxins.Conclusions: 1 Sterigmatocystin and fumonisin toxins both have cytotoxic effects alone, and with the increasing toxin concentration increased in a dose-dependent manner. 2 The combined effect of FB₁ and ST was synergistic or significant enhancement, while two kinds of toxin treated cells increased the degree of liver cell damage in rats. 3 Effects of FB₁ and ST on oxidative damage in rat liver cells.