Rizatriptan is the most popular medicine used for the treatment of acute migraine headaches and its extensive use damages the structure of deoxyribonucleic acid leading to several biological malfunctions. During its synthesis, it is possible to form nitrosamine impurity which is a potential genotoxic nature. In the present study, a new ultra-performance liquid chromatographic (UPLC) method was developed to determine Genotoxic impurity (GI) in active pharma ingredient (API) Rizatriptan benzoate and emphasizes its controlled consumption. A UPLC method was optimized for the dimer impurity 2-(5-((1H-1,2,4-triazol-1-yl)methyl)-2-((3-(2-(dimethylamino)ethyl)-1H-indol-5-yl)methyl)-1H-indol-3-yl)-N, N dimethylation-1-amine and Rizatriptan benzoate were separated on Waters Acquity BEH C18 column with dimensions of 100 mm length, 3.0 mm internal diameter and 1.8 μm of particle size by using 0.1% orthophosphoric acid in water as buffer with organic modifier acetonitrile as a gradient composition with flow rate of 1.0 ml/minute. Different column oven temperatures have been tested between the 25ºC and 45ºC temperatures and we found that the 40ºC column temperature was best for the well-separated and reproducible results. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines. The primary standard solution of dimer impurity 2-(5-((1H-1,2,4-triazol-1-yl)methyl)-2-((3-(2-(dimethylamino) ethyl)-1H-indol-5-yl)methyl)-1H-indol-3-yl)-N,N dimethylation-1-amine was prepared by dissolving 3.8 mg into a 100 ml volumetric flask. It was further diluted to make 0.38 μg/ml of working standard solution was prepared. The experiment was carried out for six replicates of 5 μl of this solution was injected into the UPLC system to check the system’s suitability. The observed results are % RSD and Tailing factors are 1.9724, and 1.12, respectively. The validation results show linearity with a correlation of coefficient values not less than 0.99. The proposed method was validated according to ICH guidelines in the 5–28 μg/g concentration range. The UV absorption maximum of dimer impurity-A was observed at 280.7 nm. The proposed UPLC technique is significant concerning the accuracy, simplicity and sensitivity for the determination of potential genotoxic dimer impurity 2-(5-((1H-1,2,4-triazol-1-yl)methyl)- 2-((3-(2-(dimethylamino)ethyl)-1H-indol-5-yl)methyl)-1H-indol-3-yl)-N, N dimethyletha n-1-amine in Rizatriptan benzoate as per API during its manufacturing. This method is reliable to use for the identification of potential GI in commercially viable drugs and pharmaceutical industries.
Key words: : Ultra Performance Liquid Chromatography; Rizatriptan benzoate; Genotoxic impurity; Acquity BEH C18 column; UV absorption maximum at 280.7 nm
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