Development of chromatographic methods for thebaine detection and quantification along with some of related alkaloid derivatives

Abstract

A RP-HPLC-UVD method for qualitative analysis and quantification of thebaine was developed utilizing RP-C18 monolithic column, gradient elution with trifluoroacetic acid and formic acid in water and in acetonitrile and detection wavelength of 285 nm. The effects of the relevant chromatographic con-ditions were investigated and the optimized and validated method was used for analysis of three in-house prepared batches of thebaine. Additionally, a complementary GC-MS method was developed for identifi-cation of thebaine and potential impurities. In parallel the batches were analyzed by routine methods for preliminary quality assessment based on physical and spectroscopic properties (melting point, specific optical rotation, UV-Vis and IR spectroscopy). There was correlation between the samples purity as de-termined by the developed HPLC method and the melting point ranges. The HPLC method herein presented is suitable for routine quantitative analysis regardless of the method of thebaine preparation and it is suitable for its corresponding free base or salt form. Additionally, considering the mobile phase com-patibility, it can be adapted/transferred easily for HPLC-MS analysis, especially if the identity of the im-purities needs to be determined.