A Novel and Sustainable HPLC Method for Monitoring Mutagenic Impurity in Injectable Medicines and Raw Ingredients via Microscale Preparation: Assessment via Greenness Tools

Abstract

The goal of the presented methodology was to evolve and verify a simple, environmentally friendly, white HPLC stability-indicating technology that can identify the mutagenic contaminant allyl bromide in both raw materials and parenteral rocuronium bromide medications. The mobile phase composition was ethyl alcohol and double-distilled water in a ratio of 35%:65% at a flow rate of 1.0 mL/min. The linearity range concentrations of allyl bromide were 1.25-10 μg/mL. The injectable volume was 20 μL, and the injection run time and total run time were less than 9 min. With a retention time of allyl bromide at 7.11 min, the detection wavelength was 210 nm. The method's BAGI degree was 77.5, the MoGAPI degree was 82 with seven green sections, the final AES degree was 87, the AGSA degree was 80.56, the CaFRI degree was 86, the CACI degree was 77, and the whiteness score was 97.1. The HPLC method features simple implementation, environmental sustainability, and compliance with ICH Q2(R2), ICH M7, and US FDA validation criteria. It offers advantages like shorter retention time, reduced reagent use, microscale sample preparation, and lower costs compared to complex systems like UPLC-MS.