During the defatting of lemuru fish in the canning production, about 15-20% of fish oil with low quality is produced. This condition demands a refinement process. Our study investigates the impact of charcoal particle size and concentration in the refining process of Lemuru fish oil. The method involved using activated charcoal with particle sizes of 177 µm and 248 µm at concentrations ranging from 2.5% w/v to 10% w/v in the bleaching process. The refined fish oil was further processed into margarine, and its stability was tested over three months at varying temperatures (5°C, 15°C, and 25°C). The research indicated that activated charcoal adsorbents in the size of 177 μm provide better results than 248 μm in terms of free fatty acid content, peroxide value, p-anisidine, and TOTOX, and thus has the potential as margarine raw material. The margarine made from the refined oil has a massive lauric acid and myristic acid. This study concludes that the particle size and concentration of charcoal significantly impact the refining process of Lemuru fish oil. The underutilized fish oil from the Lemuru canning industry shows potential as a valuable by-product for producing nutrient-rich margarine, offering a more innovative approach to support sustainable fisheries.