• Written by Gianpaolo Rota, Application Specialist | 17 September 2022

Frequently Asked Questions on microplastic determination


DETERMINATION OF MICROPLASTIC CONTENT IN SEAFOOD AND IN IN BEACH SAND SAMPLES

The increasing use of plastics in our daily lives and in manufacturing processes often culminates in a slow degradation of the materials and the subsequent contamination of our environment and food chain. Scientists define microplastics as plastic fragments less than 5 mm, or about 0.2 inches, in diameter. Nanoplastics are even smaller, with diameters less than 0.001 mm. Recent studies demonstrated that the human ingestion of MPs is about 120 thousand particles annually, which comes from water, food or even air. Research in wildlife and animal models has linked micro- and nanoplastic exposure to infertility, inflammation and cancer, but health outcomes in people are currently under assessment, with effects mainly associated with the blockage of the digestive tract. While regulators are trying to understand the extent and toxicity of the problem, researchers and analytical methods bodies are working towards standardized analytical solutions to best characterize these particles.
  1. Frequently asked questions answered by two experts in the field, prof. Cezar A Bizzi of University of Santa Maria, and prof. Jacopo La Nasa, of the University of Pisa.

Questions and Answers

Q1: How important is sample pre-treatment before Py-GC/MS? Do proteins and related tissues degrade before the polymers?
The sample pretreatment is very important before the Py-GC-MS analysis. One of the biggest problems for the protein is related to the study of polyamides. In fact, the sample pretreatments used for the digestion of polyamides could affect also these polymers, leading to a depolymerization and consequent loss of information on the sample. For this reason, it is important to define first which material you want to study and then define the right sample pretreatment considering all the possible effects on the polymers that you want to analyze.
Q2: Would the microwave assisted extraction work for other types of plastics, like PET, PVC, PA, PMMA and rubbers NR and SBR?
In the conditions that we used in this preliminary study we can also recover, together with PS, PE and PP, polycarbonate, and polyvinyl chloride. To extend the type of polymers that can be extracted/solubilized we are currently testing other types of solvents.
Q3: For GC-MS quantification, What matrix do you use for density separation? SPT? NaI? Will the digestion temperature digest PMMA? Can compost work on GC-MS for MPs determination?
In this study we did not use any density separation step, we performed directly the analysis on the sand. The extraction did not entailed any digestion, but we are working on the implementation of different solvents to extend the types of polymers that can be analyzed with this approach
Q4: Have you considered how bioplastics will be affected by either the solvent extraction/digestion. Do you think they will dissolve alongside other organic matter?
We are currently working on bioplastics, in fact these polymers are characterized by a compostable portion that solubilize easily, while the polyester portion of the material must be treated differently with proper more strong conditions
Q5: Did you find a significant difference in the quantification of MP by weigh? Do you considered quantification by counting?
The proposed methos was developed for replacing the counting step. Since we know the initial mass of MP, it was possible to evaluate and quantificate possible losses.
Q6: Did you check if there are only microplastics left on the filter, no other material?
Considering the optimized sample digestion conditions, the final solution was completely free of solid particles from the organic sample (fish tissue). The only solid material retained in filter was the particles from MPs.
Q7: Did you check the recovery by adding one size of microplastics, or a size range? What was the smallest size tested?
Three different MP size range were evaluated: 0.3-0.8 mm; 0.81-1.69 mm; and 1.7-5 mm. All the different ranges behaved in the same way.
Q9:Is the proposed method addressed also for saponification issue, especially for fatty samples?
Considering the results obtained for dissolved carbon content in the solution from digestion, it is possible to assume that all the organic sample (including fat) was, at least, partially oxidized. Saponification was not observed or explored. But, the amout of fat in sample would impair the digestion efficiency (not observed for the evaluated samples)
Q10: Did you maybe check if there were some changes on the MP surfaces after the digestion?
Taking into account the used temperature (200 ºC), they were partially melted at the end of the process (but without significative losses in the MP mass)
Q11: Will the high temperature be able to digest MPs too?
The digestion conditions (acid concentration, temperature, and reaction time) were optimized to avoid MPs degradation or solubilization
Q12: How do you work on recovery experiment with spike samples? Directly add to your samples?
MPs and fish samples were mixed previous to the digestion step but without significant changes in the fish sample and no modification in the MP particle size/shape.
Q13: Only with HNO3 the fish material will digest?
If concentrated acid is used, for sure. But, using diluted HNO3 solution (< 3 mol/L), at temperatures below 200ºC, the MPs were saved
Q14: Have you used a microplastic CRM material to evaluate the MW digestion procedure?
Not in the optimizations presented, which resulted in the STOTEN (Journal) publication. After that, we joined a collaborative work for producing a CRM of MP in fish tissue.
Q15: Is the proposed method addresses saponification issue, especially for fatty samples?
Considering the results obtained for dissolved carbon content in the solution from digestion, it is possible to assume that all the organic sample (including fat) was, at least, partially oxidized. Saponification was not observed or explored. But, the amount of fat in sample would impair the digestion efficiency (not observed for the evaluated samples)
Q16: Have you considered how bioplastics will be affected by either the solvent extraction/digestion. Do you think they will dissolve alongside other organic matter?
Bioplastics were not evaluated. I think the dissolution of them could be avoided by proper operational optimization.
Q18: Which are the best conditions to degrade MP's?
The use of higher conditions of temperature, combined with concentrated reagents.
Q19: Are you planning to evaluate your method in water samples like in fresh water and marine water?
This aspect was not considered up to this moment. I think it might work.

Guidelines for sample preparation and analysis for microplastic determination

Sample acid digestion was performed with two types of system based on different microwave technologies.
ETHOS UP

ETHOS UP

The ETHOS UP is a flexible and high performing platform used for elemental analysis. Equipped with easyTEMP contactless sensor, it directly controls the temperature of all samples and solutions, providing accurate temperature feedback to ensure complete digestion in all vessels and high safety. ETHOS UP works with SK-15 rotor capable of high temperature (up to 300 °C) and pressure (up to 100 bar). The SK-15 also features Milestone’s patented “vent-and reseal” technology for controlling the internal pressure of each vessel. This ensures complete, safe and reproducible digestions of even the most diffi cult samples.
ultraWAVE

ultraWAVE

The ultraWAVE, developed and patented by Milestone, with Single Reaction Chamber (SRC) technology utilizes high-performance stainless steel, allowing to reach higher pressures and temperatures (up to 199 bar and 300°C respectively) and to use any type of acids. Disposable vessels eliminate the need to assemble, disassemble or clean between processing. Just as important, dissimilar samples can be processed simultaneously using any mixture of disposable glass, quartz or TFM vials, thus saving time and money. The ultraWAVE is simply the fastest, easiest and most efficient digestion system ever made.