A selective and sensitive LC-MS/MS method for the simultaneous determination of twopotential genotoxic impurities in celecoxib

Abstract

Abstract Background Impurity profiling is now receiving critical attention from regulatoryauthorities. For trace level quantification of potential genotoxic impurities(PGIs), conventional analytical techniques like high-performance liquidchromatography (HPLC) and gas chromatography (GC) are inadequate; consequently,there is a great need to apply hyphenated analytical techniques to developsensitive analytical methods for the analysis of pharmaceuticals. Methods A selective and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS) method was developed for the simultaneous determination of(4-sulfamoylphenyl)hydrazine hydrochloride (SHH) and(4-methyl-acetophenone) para -sulfonamide phenylhydrazine hydrochloride(MAP) PGIs in celecoxib active pharmaceutical ingredient (API). The LC-MS/MSanalysis of SHH and MAP PGIs was done on Symmetry C18 (150 mm ×4.6 mm, 3.5 μm) analytical column, and the mobile phase used was5.0 mM ammonium acetate-acetonitrile in the ratio of 30:70( v / v ). The flow rate used was 0.7 mL/min. Triplequadrupole mass detector coupled to positive electrospray ionization operated inmultiple reaction monitoring (MRM) mode was used for the quantification of SHH andMAP PGIs. The method was validated as per International Conference onHarmonization (ICH) guidelines and was able to quantitate both SHH and MAP PGIs at1.0 ppm with respect to 10 mg/mL of celecoxib. Results The proposed method was specific, linear, accurate, precise, and robust. Thecalibration curves show good linearity between the concentration range of 0.06 and7.5 ppm for both SHH and MAP PGIs. The correlation coefficient obtained was>0.9998 in each case. The method has very low limit of detection (LOD) andlimit of quantification (LOQ). The obtained LOD and LOQ values were 0.02 and0.06 ppm, respectively, for both SHH and MAP PGIs. For both the PGIs,excellent recoveries of 95.0% to 104.0% were obtained at a concentration range of0.06 to 3.0 ppm. The developed method was also applied to determine the SHHand MAP PGIs in three formulation batches of celecoxib. Conclusions The proposed method is simple and accurate and is a good quality control tool forthe simultaneous quantitative determination of SHH and MAP PGIs at very low levelsin celecoxib during its manufacturing.