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An efficient and gentle enzymatic digestion protocol for the extraction of microplastics from bivalve tissue
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A novel tissue digestion protocol using pancreatic enzymes and a pH buffer was developed for extracting microplastics from bivalve tissue, achieving 97.7% tissue removal overnight without damaging four plastic polymers or six textile fiber types as verified by FTIR. The method provides a faster, gentler, and more polymer-compatible alternative to existing chemical digestion protocols for bivalve microplastic analysis.
Standardized methods for the digestion of biota for microplastic analysis are currently lacking. Chemical methods can be effective, but can also cause damage to some polymers. Enzymatic methods are known to be gentler, but often laborious, expensive and time consuming. A novel tissue digestion method with pancreatic enzymes and a pH buffer (Tris) is here presented in a comparison to a commonly applied digestion protocol with potassium hydroxide. The novel protocol demonstrates a highly efficient removal of bivalve tissue (97.7 ± 0.2% dry weight loss) already over-night. Furthermore, it induces no impairment in terms of ability to correctly identify four pre-weathered plastic polymers and six textile fiber polymers by Fourier transform infrared spectroscopy after exposure. The high-throughput protocol requires minimal handling, is of low cost and does not pose risk to the performer or the environment. It is therefore suggested as a candidate for a standardized digestion protocol, enabling successful analysis of microplastics ingested by bivalves.
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Researchers compared several existing methods for extracting microplastics from bivalve tissue, including hydrogen peroxide, enzymatic digestion, and potassium hydroxide treatments. They found that only the KOH method allowed filtration through very fine filters down to 1.2 micrometers when a neutralization step was added, enabling recovery of much smaller microplastics. The study recommends KOH digestion as the most practical extraction method for studies aiming to assess human exposure risk from consuming shellfish.
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