A novel and eco-friendly UPLC-ESI-MS method for the quantification of Aceclofenac-NDSRI (Nitroso Drug Substance Related Impurity) from Aceclofenac drug substance and combination formulations

Nitrosamine drug substance-related impurities (NDSRIs) have proven to be an overwhelming challenge to pharmaceutical manufacturers and regulatory bodies. In the current study, Aceclofenac(ACF)-NDSRI was synthesized by nitrosation of ACF drug substance, isolated using column chromatography, and its genotoxicity was assessed by (Q)-SAR tools. A simple, accurate, and highly sensitive UPLC-MS/MS method was developed for the quantification of ACF-NDSRI from ACF drug substances and combination formulations. The chromatographic resolution was accomplished using an Agilent Zorbax SB-C18 column (150 mm × 4.6 mm, 3.5 μm) in an isocratic elution mode with 0.1% Formic acid in a mixture of water and Acetonitrile in the ratio of 50:50 v/v, as mobile phase at a pump flow of 0.5 mL/min. Mass analysis was accomplished utilizing Xevo TQ-XS mass spectrometer with electrospray ionization in the positive mode. ACF-NDSRI quantification was carried out using multiple reaction monitoring mode as the mass transition: m/z 383.06 → 353.06. The methodology was validated successfully, in accordance with ICH guidelines and is proficient in quantifying at level 1/20th of the specification limit (0.30 ppm). The validated method revealed excellent linearity over the concentration range of 0.30–15 ng. The Pearson correlation coefficient of ACF-NDSRI was >0.999. The %RSD from method and intermediate precision were determined to be 0.96% and 1.12%, respectively; the %recovery values ranging from 91.89 to 99.14%. There were no matrix interference peaks detected during the entire chromatography run, which indicates the method is specific. In addition, the stability study of ACF-NDSRI in the sample solution was conducted and found to be stable for at least 48 Hrs. Enhanced tools, such as GAPI, AGREE, and analytical eco scale, have been used to establish the greenness and eco-friendliness of the current method. The current method could be used to quantify ACF-NDSRI in drug substance, as well as combination formulations during commercial release and stability testing. [Display omitted] •Currently, NDSRIs are a challenge to the pharma industry and regulatory bodies.•Synthesis and mutagenicity evaluation of Aceclofenac (ACF)-NDSRI by (Q)-SAR tools.•Acceptable intake of ACF-NDSRI was calculated using recent regulatory guidelines.•UPLC-MS Method greenness was evaluated by GAPI, AGREE and Analytical Eco-scale.•This novel method can be used to quantify ACF-NDSRI with a shorter run time.