Manchuri Krishna Moorthy, Shaik Mahammad Ali, G. Reddy
Jan 12, 2022
Citations
0
Influential Citations
4
Citations
Journal
Biomedical chromatography : BMC
Abstract
The foremost objective of the present study is to develop and validate a new LC-QTOF-MS/MS method for identification and quantitative determination of 4,6-dichloro-5-nitro-2-(propylthio) pyrimidine (DPP) genotoxic impurity through derivatization process in ticagrelor active pharmaceutical ingredient (API). Due to low response of DPP at specification level, the DPP is converted to 4,6-dibenzylamino-5-nitro-2-(propylthio) pyrimidine (DPP derivative) by addition of benzyl amine, then analyzed using mass spectrometry with a time-of-flight analyzer and accomplished good separation under the experimental conditions described. The effective separation of DPP derivative was achieved using ACQUITY UPLC BEH C18 reverse phase column (100 mm x 4.6 mm x 1.7 μm) with an isocratic program wherein mobile phase-A used as 0.1% formic acid in milli Q water and mobile phase-B used as acetonitrile in the ration of 20:80 v/v. Flow rate maintained 0.4 mL/min, injection volume 2 μL, auto sampler temperature 5°C, column oven temperature ambient and run time 6.0 minutes. Diluent used as 0.2% benzyl amine in water and acetonitrile in the ratio of 30:70 v/v. Retention time of DPP derivative was found at 2.87 minutes. Limit of detection (LOD) and limit of quantification (LOQ) were found at 0.03 ppm and 0.08 ppm, respectively. The DPP derivative was linear from 1.68 ppm to 12.78 ppm with R2 of 0.9958. Thus, the developed method is valid for identification and quantitative determination of 4,6-dibenzylamino-5-nitro-2-(propylthio) pyrimidine (DPP derivative) in ticagrelor API.