NUCLEPAR – Specially manufactured nucleic acids marked nanoplastics for water technology applications using Electrohydrodynamic Atomization (EHDA)

Environmental nano- and micro-plastics (NMPs) are highly diverse [2]. Accounting for this diversity is one of the main challenges to develop a comprehensive understanding of NMPs detection, quantification, fate, and risks [3]. Two major issues currently limit progresses within this field: (a) validation and broadening the current analytical tools (b) uncertainty with respect to NMPs occurrence and behaviour at small scales (< 20 micron). Tracking NMPs in environmental systems is currently limited to micron size plastics due to the size detection limit of the available analytical techniques. There are currently many uncertainties regarding detecting nanoplastics in real environmental systems, e.g. the inexistence of commercially available NMPs and incompatibility between them and those generated from plastic fragments degradation in the environment. Trying to tackle these problems some research groups synthesized NMPs dopped with metals inside [16]. However, even though elemental analysis techniques (ICP-MS) are rather sensitive, the low volume of these metals encapsulated in the nanoparticles make their detection rather challenging. At the same time, due to Sars-Cov-19 pandemic, nucleic acid identification technologies (LAMP, PCR) experienced a fast evolution and are able to provide detection at very low levels with very compact and reliable equipment. Nuclepar proposes the use of Electrohydrodynamic Atomization (EHDA) to generate NMPs coated with nucleic acids of different polymer types, sizes, and shapes, which can be used as support for detection of such particles using PCR-LAMP technology. If proven possible, Nuclepar might become a first step towards an easy NMPs detection tool. This knowledge will certainly impact current risk assessment tools, efficient interventions to limit emissions and adequate regulations related to NMPs.

Environmental nano- and microplastics (NMPs) are highly diverse, posing challenges for their detection,
quantification, and risk assessment. Two key limitations are the lack of validated analytical tools and the
difficulty in tracking NMPs smaller than 20 μm. Nanoplastics are especially hard to study due to the absence
of commercial standards and poor compatibility with naturally degraded fragments. While some studies use
metal-doped NMPs for detection, their low metal content limits sensitivity. Meanwhile, nucleic acid detection
methods such as LAMP-PCR have advanced significantly during the COVID-19 pandemic, offering high
sensitivity in compact formats. Fabrication of nucleic acid-doped NMPs presents a promising strategy to
enhance nanoplastics research through PCR-LAMP detection. Nuclepar proposed employ
Electrohydrodynamic Atomization (EHDA) also known as electrospray, to produce nucleic acid-doped
micro/nanoplastic particles (NMPs), enabling their detection through PCR-LAMP technology. In this project,
the OmniLAMP device developed by Visuri was used for LAMP PCR testing to detect nucleic acid-marked
NMPs. Originally designed for molecular pathogen detection, this portable, low-cost device uses colorimetric
Loop-Mediated Isothermal Amplification to amplify genetic material. A resulting color change indicates the
presence of the target, which is automatically captured and analyzed by the device’s integrated software. Using
the EHDA technique, nucleic-acid dope NMPs (polystyrene particles) with a target size of 1 μm were
successfully produced. Key parameters, such as flow rates, collection time, nozzle-to-collector distance, and
applied voltage, were optimized. Particles, with and without DNA, were tested via OmniLAMP, showing a pinkto-yellow color change in the presence of target DNA. These results highlight the potential of nucleic-acid
doped particles as molecular tracers, using the LAMP-PCR technique, particularly with the OmniLAMP device,
to support the development of advanced tools for monitoring and removing NMPs in wastewater and
environmental systems. AND