Methods for controlled preparation and dosing of microplastic fragments in bioassays

Microplastic fragments (microfragments) are among the most abundant microplastic shapes found in marine ecosystems throughout the world. Due to their limited commercial availability, microfragments are rarely used in laboratory experiments. Here a novel method of microfragment production has been developed and validated. Polyethylene and polypropylene plastic stock (2 and 3 mm thick respectively) was ground using a cryomill, washed, and rinsed through a stack of sieves. Microfragments were prepared at three distinct size classes (53-150, 150-300, 300-1000 μm) and were confirmed to be accurate and consistent in size. Employing a novel ice cap dosing technique, microfragments were accurately dosed into experimental vials while excluding headspace, facilitating particle suspension without the aid of chemical surfactants. A proof of principle ingestion experiment confirmed the bioavailability of 53-150 μm polyethylene microfragments to brine shrimp Artemia sp. Together, these methods provide a controlled way to produce and dose microplastic fragments for experimental and analytical research.