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Internal combustion engines, as the dominant power source in the transportation sector and the primary contributor to carbon emissions, face both significant challenges and opportunities in the context of achieving carbon neutral goal. Biofuels, such as biodiesel produced from biomass, and zero-carbon fuel ammonia, can serve as alternative fuels for achieving cleaner combustion in internal combustion engines. The dual-fuel combustion of ammonia-biodiesel not only effectively reduces carbon emissions but also exhibits promising combustion performance, offering a favorable avenue for future applications. However, challenges arise in the form of unburned ammonia (NH3) and N2O emissions. This study, based on a ammonia-biodiesel duel-fuel engine modified from a heavy-duty diesel engine, delves into the impact of adjustments in the two-stage injection strategy on the combustion and emission characteristics.