Quantitative Determination of 5 Nitrosamines in Moxifloxacin by UHPLC-HRMS

OBJECTIVE To develop and validate a UHPLC/HRMS method for simultaneous quantitation of 5 genotoxic nitrosamines in moxifloxacin.METHODS ACE Excel 1.7 C₁₈-PFP(100 mm×2.1 mm, 1.7 μm) column was used, separated by gradient elusion using a mixture of water and methanol with 0.1% formic acid at 0.3 mL·min^-1, and 5 nitrosamines were detected by parallel reaction monitoring using Q Exactive mass spectrometer in electrospray positive mode.RESULTS The lower limits of quantitation for 5 genotoxic nitrosamines ≤0.5 ng·mL^-1. Good linearity was established for NPYR, NEMA and NDEA over 1.0-100 ng·mL^-1, and for NPIP and NDBA over 0.1-100 ng·mL^-1(R²≥0.998), respectively. The intra-assay accuracy of 5 nitrosamines in moxifloxacin hydrochloride was determined to be from 93.1% to 114.1%, and the RSD was found to be from 0.7% to 4.6%.CONCLUSION The validated method provides satisfactory sensitivity and specificity for analysis of genotoxic nitrosamines as impurities in moxifloxacin hydrochloride.