Isolation and identification of anthropogenic particles in fish stomachs by Raman spectroscopy: a new method

Microplastic particles (MP) contaminate oceans and affect marine organisms in several ways. Ingestion combined with food intake is generally reported. However, data interpretation is often circumvented by the difficulty to separate MP from bulk samples. Visual examination is often used as one or the only step to sort these particles. However, color, size and shape are insufficient and often unreliable criteria. Here we present an isolation method of MP specially adapted to a subsequent analysis by Raman spectroscopy. This method avoids fluorescence problems allowing the identification anthropogenic particles (AP) from stomach contents of fish by Raman spectroscopy. It was validated with commercial samples of microplastics and cotton along with stomach contents from three different Clupeiformes fishes: Clupea harengus, Sardina pilchardus and Engraulis encrasicolus. The optimized digestion and isolation protocol showed no visible impact on microplastics and cotton particles while the spectroscopic analysis allowed precise identification of microplastics and textile fibers. This approach allowed us to isolate 35 particles. These were analyzed by Raman spectroscopy: eleven were microplastics and thirteen were made of cellulose or lignin, or both (mostly fibers). Some particles were not identified but contained artificial colorants. This isolation protocol will help to assess the presence, quantity and composition of AP in planktivorous fish stomachs.