Quantitative Analysis Method Research Of Free Gossypol In Animal Rerived Food

In this paper the determination methods of free gossypol concentration in cottonseed and animalderived food were studied. The free gossypol was split through pre-column derivatization, the HPLC-MSmethods for determination of free gossypol concentration in cottonseed, cotton stalk and leaf wereestablished. The HPLC-MS methods for determination of (-)-gossypol and (+)-gossypol in cottonseed werebuilt. Based on that, the HPLC methods for determination of (-)-gossypol and (+)-gossypol in rat plasmawere established, the HPLC-MS methods for determination of (-)-gossypol and (+)-gossypol in rat plasma,liver and muscle were further established. The (-)-gossypol and (+)-gossypol concentration in animalderived food, such as chicken liver, pigs liver, chicken and pork, which were purchased from markets inUrumqi, were detected by the established methods. The results are as follows:1. The free gossypol concentration in cottonseed, cotton stalk and leaf were detected by HPLC-MSinternal standard method with glycyrrhetic acid as internal standard. Results showed: the free gossypol incottonseed, cotton stalk and leaf were6.66~10.14mg/g,0.018mg/g,0.015mg/g respectively. The HPLC-MSinternal standard methods established are sensitive and specific with convenient and feasible sampletreatment method, and could be used to determined accurately the free gossypol concentration in cottonseed,cotton stalk and leaf.2. The content of (-)-gossypol and (+)-gossypol in cottonseed were determined by HPLC-MS methodusing pre-column derivation with (R)-(-)-2-Amino-1-propanol ast he chiral separation reagent. Resultsshowed: the (-)-gossypol and (+)-gossypol in cottonseed were1.60~4.09mg/g,2.30~5.69mg/g respectively.The method was higher sensitivity and lower interference of dopant, and can provide valuable preferencesfor detecting the optical isomers of gossypol in the biological sample.3. The contents of (-)-gossypol and (+)-gossypol in rat plasma were determined by HPLC methodusing pre-column derivation. The sample was treated with acetonitrile to remove protein and extract thefree gossypol, then the free gossypol was concentrated by the nitrogen flow. Being simply and usable,methodological study of the method was conducted, which could be used to determine the content of(-)-gossypol and (+)-gossypol in blood plasma. Resulted showed: the content of (-)-gossypol and(+)-gossypol in flood plasma of rat fed with drug were0.0767~0.1318μg/mL and1.5213~1.7417μg/mLrespectively.4. The contents of (-)-gossypol and (+)-gossypol in rat were determined by HPLC-MS internalstandard method using pre-column derivation. Results showed: the free gossypol were accumulated in ratplasma, liver and muscle with different levels, the content of free gossypol in liver are higher than plasmathan muscle. However, the (+)-gossypol concentration was higher than that of (-)-gossypol, especially inflood plasma. The method established can determine free gossypol in animal derived food5.The content of free gossypol in different animal derived food which were bought from two maketwere detected using this method. The result showed animal livers was low, some of which did not reach thedetection limit, the free gossypol was not detected in animal muscle. There are seventeen chicken liversamples, thirteen samples have free gossypol, the content of (-)-gossypol is0.0337~0.6385μg/g, the contentof (+)-gossypol is0.0753~0.5664μg/g, Eight pig liver samples, and seven samples have free gossypol, thecontent of (-)-gossypol is0.0626~0.7713μg/g, while (+)-gossypol is0.0301~0.4392μg/g. Six fish liversamples, three have free gossypol the content is lower, the content of (-)-gossypol is0.0316~0.0510μg/g,while (+)-gossypol is0.0342~0.0737μg/g.