A selective and sensitive novel methods were developed and validated for identification and trace level quantification (0.5 ppm) of genotoxic impurities Methyl 3‐amino‐4‐methylbenzoate, 3‐amino‐4‐methylbenzoic acid, and 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline in Nilotinib dihydrochloride active pharmaceutical ingredient by using liquid chromatography‐tandem mass spectrometry. The developed and validated methods were more accurate and capable to confirm the m/z values of parent and fragment ions through mass spectrometry and tandem mass spectrometry for further fragmentation. In addition, the developed methods were validated through current regulatory guidelines with a lower detection level of 0.15 ppm and a quantification level of 0.5 ppm for all the three genotoxic impurities. The correlation coefficient was observed as 0.9997 for Methyl 3‐amino‐4‐methylbenzoate genotoxic impurity‐I, 0.9998 for 3‐amino‐4‐methylbenzoic acid, and > 0.9999 for 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline. The recovery percentage of all the three genotoxic impurities was found to be between 93% to 105%.

A selective and sensitive novel method was established and validated for well-characterized tert-butyl ((1S, 3R)-3- acetylcyclopentyl) Carbamate (Acetyl BOC) and tert-butyl (1S, 3R)-3-hydroxycyclopentyl) Carbamate (Hydroxy BOC) genotoxic impurities in Bictegravir sodium active pharmaceutical ingredient by using liquid chromatographytandem mass spectrometry. The developed method was more accurate and capable to confirm the m/z values of parent and fragment ions, method was validated through ICH guidelines to detect 0.3 ppm and estimate up to 1.0 ppm of both impurities. The correlation coefficient was observed > 0.9993 and the recovery percentage of genotoxic impurities was found to be between 93% to 108%.

A new rapid run time LC-MS method was developed and validated for detection (0.03 ppm) and quantification (0.1 ppm) in ultra-trace level of genotoxic impurity (GTI) ethyl-(1R,5R,6R)-7-(tertbutyl)- 5-(pentan-3-yloxy)-7-azabicyclo[4.1.0]hept-3-ene-3-carboxylate in oseltamivir phosphate active pharmaceutical ingredient (API). The method is price effective, time redeemable and capable to confirm the parent and daughter ion masses through mass spectrometry and tandem mass spectrometry for further fragmentation. An isocratic program and YMC Pack Pro C4 reverse phase column (150 mm × 4.6 mm × 3.0 μm) was used to achieve separation between oseltamivir phosphate and ethyl-(1R,5R,6R)- 7-(tert-butyl)-5-(pentan-3-yloxy)-7-azabicyclo[4.1.0]hept-3-ene-3-carboxylate impurity. Mobile phase-A used was 0.01 M ammonium acetate in water and mobile phase-B used was acetonitrile in the ration of 40:60 v/v. Diluent was used methanol. The chromatographic conditions were used, injection volume: 20 μL, flow rate: 1.0 mL/min, oven temperature: 50 ºC, auto sampler: 5 °C and run time 8.0 min. The detection and quantification levels found at 0.03 and 0.1 ppm, respectively. Ethyl (1R,5R,6R)- 7-(tert-butyl)-5-(pentan-3-yloxy)-7-azabicyclo[4.1.0]hept-3-ene-3-carboxylate impurity is linear from 0.1 to 15 ppm levels with regression coefficient 0.9994. The recoveries were found in the range of 93.8% to 105.0%.