Protocol for extraction and analysis of microplastics in freshwater, sediment, and fish samples

Microplastic pollution threatens aquatic ecosystems and food webs. Here, we present a protocol for extracting microplastics from freshwater, sediment, and fish digestive system samples. We collect water using manta nets, sample sediment with benthic grabs, and dissect fish to isolate digestive organs. We perform chemical digestion, density separation, vacuum filtration, and microscopic enumeration, followed by polymer identification using μ-Raman spectroscopy. Quality control steps ensure high recovery and minimize contamination. For complete details on the use and execution of this protocol, please refer to Gündoğdu et al.^1,2.