Use of 2H-labeled nanoplastics to study their accumulation and toxicity in Daphnia magna.

Roxane Danquigny, Bruno Grassl, Séverine Le Faucheur, Javier Jiménez-Lamana, Marc Metian, Stéphanie Reynaud
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Abstract

Understanding the interactions between nanoplastics (NP) and biota is essential for risk assessment. However, NP quantification in complex matrices remains a challenge, especially when they are not labeled. Most labeling strategies consists in adding another compound (fluorophore, metal, lanthanide…) to the polymer, which can alter the NP properties and poses a risk of leaching phenomena. In the present study, we synthesized spherical and monodisperse 2H-labeling NP (129 nm) with carboxyl groups at their surface (8.7 nm-2) using surfactant-free polymerization. A pyrolysis coupled with gas chromatography/mass spectrometry method was developed to enable their quantification (limit of detection = 7.4 ng), without extensive sample preparation or significant matrix effects. The 2H-labeled NP were then used to study their toxicity and accumulation in the planktonic crustaceans Daphnia magna (D. magna). Daphnids were exposed for 48 hours to the deuterated NP at different concentrations, including environmentally realistic ones (from 0.03 to 36 mg kg-1). Little to no acute toxicity were observed in this range of concentration. During the experiment, adsorption on polycarbonate well walls was observed, suggesting that the NP concentration during exposure experiments with organisms should be monitored to take into account possible loss. The NP accumulation (mainly in the digestive tract) increased with the exposure concentration, tending to a plateau at higher concentrations. The depuration was also investigated and was significantly higher (97 % against 59 % in average) when the daphnids have access to algae, which highlight the need to add food in D. magna depuration studies to study the NP fate in these organisms. The present study demonstrated the advantages of stable isotope-labeled NP to better understand the processes controlling NP accumulation and impacts on aquatic biota.

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使用 2H 标记的纳米塑料研究其在大型蚤体内的积累和毒性。
了解纳米塑料(NP)与生物群之间的相互作用对于风险评估至关重要。然而,复杂基质中 NP 的定量仍然是一项挑战,尤其是在没有标记的情况下。大多数标记策略都是在聚合物中添加另一种化合物(荧光团、金属、镧系元素......),这会改变 NP 的特性,并带来浸出现象的风险。在本研究中,我们采用无表面活性剂聚合法合成了表面带有羧基(8.7 nm-2)的球形单分散 2H 标记 NP(129 nm)。所开发的热解耦合气相色谱/质谱法可对其进行定量(检测限 = 7.4 ng),而无需大量的样品制备或显著的基质效应。然后,利用 2H 标记的 NP 来研究它们在浮游甲壳动物大型水蚤(D. magna)体内的毒性和积累。将水蚤暴露在不同浓度的氚代 NP(包括符合环境条件的浓度(0.03 至 36 毫克/千克))中 48 小时。在此浓度范围内,几乎没有观察到急性毒性。在实验过程中,观察到聚碳酸酯井壁上有吸附现象,这表明在对生物进行暴露实验时,应监测 NP 浓度,以考虑到可能的损失。NP 的积累(主要在消化道中)随着暴露浓度的增加而增加,在浓度较高时趋于平稳。研究还调查了水蚤的净化情况,当水蚤能接触到藻类时,净化率明显更高(97%,而平均净化率为 59%),这突出表明有必要在水蚤净化研究中添加食物,以研究 NP 在这些生物体内的归宿。本研究证明了稳定同位素标记的 NP 在更好地了解 NP 累积过程及其对水生生物群影响方面的优势。
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