Detection Of Gossypol And Its Metabolite Gossypolone In Animal Derived Food

In this paper, the metabolites of gossypol in chicken and crucian were preliminarily studied, and the gossypolone of gossypol metabolites was prepared. The LC-MS/MS methods for determination of free gossypol, gossypolone,(+)-gossypol and(-)-gossypol in chicken liver were established. The sampling survey was conducted to determinate the contents of free gossypol, gossypolone,(+)-gossypol and(-)-gossypol in different animal derived foods. The main results are as follows:(1) Samples of blood plasma and liver of chicken and crucian were analyzed by ESI-QTOF/MS to find gossypolone of gossypol’s oxidization product. The molecular formula and molecular weight of gossypolone were C30O26H11 and 546.1526, and its structural formula was 6,6’,7,7’-tetrahydroxy-5,5’-diisopropyl-3,3’-dimethyl-[2,2’]-binaphthyl-1,1’,4,4’-tetraone-[8,8]-dicarboxaldehyde.(2)The oxidization process of gossypol was simulated by chemical synthesis methodology to prepare the gossypolone. Then the structure of oxidization product was identified by LC-MS/MS and ESI-QTOF/MS, the purity was analyzed by HPLC. The results indicated that the molecular weight, molecular formula, structural formula of product prepared in experiment were consistent with gossypolone. It can be inferred that the oxidization product was gossypolone, and the product content was up to 95 %.(3)The contents of gossypol and gossypolone in chicken liver were determined by LC-MS/MS. Though the method ological verification and re-verification, the quantitative analysis was not interfered with the endogenetical matrices in samples, and the method has a higher linearity. All of the methodological verification indexes met basically the requirements of quantitative analysis of the guiding principles, so the method could be used in the quantitative analysis of free gossypol and gossypolone in chicken liver, chicken, fish liver, fish and other biological samples.(4)The LC-MS/MS method for the quantitative analysis of(+)-gossypol and(-)-gossypol in chicken liver was established. The quantitative analysis of samples was not interfered with the endogenetical matrices in samples, and the method has a higher linearity. All of the methodological verification indexes met basically the requirements of quantitative analysis of the guiding principles, so the method could be used in the quantitative analysis of(+)-gossypol and(-)-gossypol in chicken liver, chicken, fish liver, fish and other biological samples.(5)Ninety animal derived food samples were purchased from three farmers’ markets of Urumqi, two farmers’ markets of Fukang and two farmers’ markets of Changji, including 15 chicken liver, 15 pork liver, 15 fish liver, 15 chicken, 15 pork and 15 fish. The contents of free gossypol, gossypolone,(+)-gossypol and(-)-gossypol in ninety animal derived food were determined by LC-MS/MS. The results indicated that the contents of free gossypol in liver samples was 0.2231~1.5463μg/g, and the detection rate was 82.2%. The(+)-gossypol and(-)-gossypol in liver samples were 0.2143~0.7420μg/g and 0.2045~0.7372μg/g, respectively, with the detection rate of 71.1%, 64.4%. The gossypolone in liver samples and free gossypol, gossypolone,(+)-gossypol and(-)-gossypol in muscle samples were not detected.