Toxic Effects And Pollution Status Of Mian Gossypol Degradation Products In Cottonseed Oil

Objective:Previous studies have shown that the degradation product tetramethoxy gossypol has some toxicity on spermatocytes and can induce the mice sperm to be aberrant and genotoxic in the experimental dose.In this experiment,through self-made non-detoxified cottonseed oil and separation and purification of higher purity degradation product tetramethoxy gossypol,a more comprehensive study of the toxicity of the degradation product tetramethoxy gossypol by in vivo experiments,while testing the Xinjiang region pollution status of degradation products in selling cottonseed oil.Methods:1.Self-made non-detoxified cottonseed oil samples,the main free gossypol degradation products prepared by ODS column method for the later animals experiments;2.Using Kunming mice to investigate the acute toxicity of tetramethoxy gossypol and to provide doses for late long-term toxicity experiments;3.The long-term toxicity of tetramethoxy gossypol and its effect on rats in SD rats after intragastric administration for 30 days;4.Comprehensive assessment of the toxicity of tetramethoxy gossypol by HE staining of tissue sections,biochemical indicators of body fluids and NMR metabolomics.5.High-performance liquid chromatography was used to determine the content of tetramethoxy gossypol in 95 batches of commercially available cottonseed oil,which was used to evaluate the contamination of commercially available cottonseed oil.Results:1.The ODS was used to separate and purify the free gossypol degradation products from non-detoxified cottonseed oil.HPLC analysis showed that the purity was 85.30%tetramethoxy gossypol according to the area normalization method,which was 7.10%higher than that of the previous study;2.Acute toxicity test results:No deaths occurred after administration of tetramethoxy gossypol maximum dose of 25mg/Kg and maximum dose volume of 0.40mL/10g.Therefore,LD50 values could not be calculated,and spssl7.0 statistical software was used for small The statistical analysis of the rats' weight and organs showed significant differences in the cardiac coefficients between the mice in the administration group and the control group(P<0.01),and there was no significant difference in other organs and weight coefficients(P>0.05);3.Long-term toxicity of tetramethoxy gossypol:The statistical analysis of rats' body weight and organs was performed using spss 17.0 statistical software.The liver and spleen coefficients were significantly different between the mice in the administration group and the control group(P<0.01).There was no significant difference between other organs and weight coefficient(P>0.05);4.Results of tissue section:There were no obvious pathological changes in the heart,spleen,lung,and kidney sections of the rats in the control group and the administration group,but there were obvious changes in the liver of the rats in the administration group.It shows that long-term administration of tetramethoxy gossypol may affect the morphology of rat liver tissue;5.The results of biochemical indicators:biochemical indicators of AST,BUN,GLU,Bb were significantly different(P<0.05),other biochemical indicators had no significant difference(P>0.05).The contents of AST and BUN in the drug-administered group were lower than those in the control group,and the contents of GLU and Bb were higher than those in the control group;6.Through the NMR metabolomics test of rat tissues,urine and feces,the results showed that tetramethoxy gossypol can affect the aerobic oxidation of sugar,glycolysis and anaerobic glycolysis,amino acid synthesis and metabolism,Lipid metabolism,energy metabolism,and damage to liver tissue,causing changes in renal function,changes in the intestinal environment,causing certain inflammation,and disorders of the psycho-endocrine system,etc;7.95 batches of commercially available cottonseed oil samples were tested.Among them,tetramethoxy gossypol was detected in 54 batches of cottonseed oil samples.The content was 0.026-1.513?g/mL.Conclusion:Only by free gossypol evaluation cottonseed oil detoxification situation is not comprehensive enough,but also through the gossypol degradation products traceability of cottonseed oil production enterprises detoxification process,so as to effectively regulate,but also for cottonseed oil free gossypol degradation products testing standards laid the foundation for testing.