DOI: 10.11648/j.ajset.20261104.23 ISSN: 2578-8353

Determination of Sensitive Method for Quantification of N-Nitroso EZD3A Impurity in Enzalutamide by LC-MS/MS Method: Applications for Routine Laboratory Testing

Prasad Pasumarthi, Santosh Bindhani, Bodroth Prasad, Bhagavathula Diwakar
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Enzalutamide, an anti-androgen agent for prostate cancer, has recently been associated with the occurrence of nitrosamine impurities, which are potential carcinogens of regulatory concern. Among these, N-Nitroso EZD3A [4-((2-cyanopropan-2-yl) (2-nitroso) amino)-2-fluoro-N-methylbenzamide] has been detected in certain batches of Enzalutamide, highlighting the need for highly sensitive methods for monitoring its presence at trace levels. Current work reports the development and validation of a robust LC-MS/MS method employing a valve-switching approach for selective quantification of N-Nitroso EZD3A in Enzalutamide. The method minimized matrix interference by preventing co-elution of the parent drug into the mass spectrometer, thereby enhancing sensitivity and specificity. Validation of the method showed linearity between 0.5 and 9.4 ppm (r2 = 0.999), precision with an RSD of 15% or less at the specification level, and recovery between 70% and 130% across LOQ to 150% levels. There was no interference from Enzalutamide or Blank, which shows that the test is specific and reliable under different analytical conditions. The method acquired quantification limitations that were much lower than the regulatory threshold. On commercial batches of Enzalutamide, the method showed that the quantities of impurities varied, with some samples going over the acceptable limit of 6.25ppm. This shows that routine monitoring is needed. The developed LC-MS/MS method provides a reliable analytical tool for trace-level quantification of N-Nitroso EZD3A in Enzalutamide.

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