Study On Quality Of Enzalutamide API And Comparison Of Determination Methods For Benzene

Enzalutamide,an androgen receptor antagonist,was approved by FDA and launched in United States in August 2012 for the treatment of castration-resistant prostate cancer.In this thesis,the quality of enzalutamide API was studied for providing the basis for the formulation of its quality standard.Meanwhile,the comparison of two methods for determination of benzene was provided reference for the quantitative analysis of benzene.The research work are listed as follows.Part 1 Quality study of enzalutamide API 1)Structure confirmation,physicochemical properties,identification and general inspection item of enzalutamide API.In this thesis,the structural elucidation of enzalutamide API was determined by UV,IR,NMR spectrum(1H-NMR,13C-NMR,HSQC,HMBC and so on)and High resolution mass spectrometry.Its physical and chemical properties such as appearance,melting point and solubility were investigated.Three methods,namely UV,IR and HPLC,were established for identification.Water content,burned residue and heavy metal test were also examined and the limits were determined.2)Related substances research of enzalutamide APIIn this thesis,13 impurities which were produced or degraded in the production and of enzalutamide API were studied.Four degradation impurities were respectively identified by the LC-MS/MS method,the impurity contrast peak comparison method and the NMR spectra.Through various combinations and systematic verification,a HPLC-UV method with strong specificity,high sensitivity,accuracy and stability was finally established for the determination of related substances.At the same time,the correction factor of each impurity with respect to the main peak was also calculated,13 impurities were quantitatively determined by method of principal component self-comparison considering the correction factors.3)Residual solvent test of enzalutamide APIA GC-FID method was established for the separation and analysis of 5 kinds of residual solvents(alcohol,isopropyl alcohol,DMSO,IPAc,THF)in the enzalutamide API,which was systematically validated meanwhile.The five kinds of residual solvents were quantitatively determined by external standard method.4)Content determination of enzalutamide APIA HPLC-UV method was established for the content determination of enzalutamide API with systematical validation,and the external standard method was also used for it.And the accuracy of HPLC-UV method was verified by quantitative NMR spectra.Part 2 Comparison for the measurement of benzene by two methodsTwo methods,namely the gas chromatography tandem mass spectrometry and the headspace gas chromatography,were established for the quantitative determination of benzene residue in the raw material by taking LKSL as API model,with a systematical comparative study meanwhile.1)Establishment of gas chromatography tandem mass spectrometryBy selecting the appropriate quantitative ion and using selected ion mode,a GC-MS method was established to quantitatively determine benzene residual in raw materials,with systematical validation.The quantitative limit of method can be up to 10.7 ng/m L.2)Establishment of headspace gas chromatography methodSampling by headspace,selecting appropriate digestion solvent to improve the response values of benzene,a headspace gas chromatography was established to quantitatively determine benzene residual in the raw materials,with systematical validation.The quantitative limit of method can be up to 20.34 ng/mL.3)A comparative study of two methods for quantitative determination of benzene was carried out to determine the applicability of the method.Through the above research,systematic quality study of enzalutamide API was set up,which will lay solid foundation for its drug declaration;comparative study of two methods for benzene determination was also established,which will provide a useful reference for analysis workers to select the quantitative method of benzene.