The Study On The Related Substance Of Propyphenazone And Its Analytical Method

In the Pharmacopoeia of each country, the impurities contained in Propyphenazone are not controlled by HPLC method and nor is there detailed study of impurities. Regarding API-Propyphenazone produced by Shandong Xinhua Pharmaceutical Co., Ltd., the study of analytical method of impurity is conducted and the best chromatographic condition is found. And the methodology validation is conducted. The related substances contained in Propyphenazone are synthesized, and their chemical structures are classified. The chemical structures are confirmed for the unspecified impurities contained in Propyphenazone. Propyphenazone is sampled at random and tested to verify if the analytical method applies to the determination of related substances contained in Propyphenazone.Firstly, analytical method of related substances contained in Propyphenazone is established. Degradation experiment is made for Propyphenazone under strong acid, strong alkali, heat, light and oxidation conditions. Aiming at the impurities generated under each condition, determination conditions are optimized and selected, including chromatographic column, mobile phase, wavelength, etc to confirm the chromatographic conditions. Meanwhile, methodology validation is conducted, such as specificity, accuracy, precision, linearity and range, robustness, LoD and LoQ. Secondly, aiming at the unspecified impurities contained in Propyphenazone, chemical structure is confirmed by adopting two ways:first is HPLC method, the retention times of two unspecified impurities in the chromatograms obtained with Propyphenazone produced by Shandong Xinhua Pharmaceutical Co., Ltd totally correspond to that in the chromatogram obtained with Propyphenazone Reference Standard; Second is LC-MS hyphenated techniques. The MS spectrums of two unspecified impurities are comprehensively analyzed, meanwhile, the MS spectrums and UV spectra of the impurities are respectively compared with that of the reference standard to confirm the structure of the unspecified impurities.The best HPLC conditions are obtained through the study:Wavelength:210nm; Chromatographic column:Shim-pack CLC Cg (4.6×150mm,5μm); Mobile phases: Phosphate buffer solution0.05mol/L KH2PO4[(solvent:water:acetonitrile=60:40); pH5.2.] The validation result of the analytical method shows that the specificity, precision, linearity and range, and accuracy of this analytical method meet the requirement, sensitivity is high. The robustness tests, changing each condition, such as change in standing time of each impurity contained in Propyphenazone and change in ratio of organic phase in mobile phase, meet the requirement. The validation result also proves sample solution of Propyphenazone is very stable, and the analytical method applies to the determination of related substances.The analytical method established in this topic can better control the content of related substances contained in Propyphenazone, and can accurately determine all the impurities contained in Propyphenazone. This analytical method is of high sensitivity, simple and easy to use and accurate, and has given an important significance to the quality control of finished Propyphenazone.