A Novel UHPLC–MS/MS Method Development and Validation for Identification and Quantification of Genotoxic Impurity Bis (2-Chloroethyl) Amine in Aripiprazole Drug Substance

A selective and sensitive novel reverse-phase method was developed and validated successfully using waters acquity ultra-performance liquid chromatography H-class system coupled with waters Micromass Quattro Premier XE (UHPLC–MS/MS) for the identification and quantification of bis (2-chloroethyl) amine genotoxic impurity in aripiprazole drug substance. The developed method employs a positive-ion electrospray ionization (ESI) with multiple reaction monitoring (MRM) detection mode using ACE 3 C18 (100 mm × 4.6 mm × 3.0 µm) column. An isocratic program was developed for the rapid analysis using 0.2% formic acid (mobile phase-A) in milli Q water and methanol (mobile phase-B) in the ratio of 45:55 v/v. The elution of bis (2-chloroethyl) amine monitored using triple quadrupole mass spectrophotometer with injection volume 2 μL, column oven temperature 40 °C, auto-sampler temperature 15 °C, and flow rate 0.5 mL/min. The retention time of bis (2-chloroethyl) amine was observed at 2.00 min. The limit of detection and the limit of quantification were observed at concentration levels of 0.070 ppm and 0.206 ppm, respectively. The correlation coefficient (R2) obtained was 0.9892 and the percentage recoveries of the method was between the ranges of 92.0–111.0%. The method was successfully developed and validated according to the current requirements of International Council for Harmonization (ICH Q2 (R1) and ICH M7 (R1)) for genotoxic impurities. The developed method was sensitive, selective, rapid, accurate, and precise, and, therefore, can be used for the identification and quantification of bis (2-chloroethyl) amine genotoxic impurity in aripiprazole drug substance.