The first evidence of microplastic occurrence in mine water: The largest black coal mining area in the Czech Republic

doi:10.1016/j.watres.2023.120538

Water Research 水研究

IF 11.4JCR Q1环境科学与生态学1区TopEI

Volume 244, 1 October 2023, 120538
卷 244， 1 十月 2023 ， 120538

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https://doi.org/10.1016/j.watres.2023.120538

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Highlights 突出

•
First study about the abundance of microplastics (MPs) in mine water.
关于矿井水中微塑料（MPS）丰度的首次研究。
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MPs in mine water at depths of 622 and 709 m ranged from 2.5 to 17.5 items/L.
在622米和709米深的矿井水中，MP含量为2.5至17.5件/升。
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MPs in shallow wells at a depth of 5 m ranged from 2.5 to 20 items/L.
在5 m深度的浅井中，MPs范围为2.5至20个项目/升。
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Most likely, microplastics have polluted all water sources, including mine water.
最有可能的是，微塑料已经污染了所有水源，包括矿井水。

Abstract 抽象

Climate change is creating new challenges for water supply worldwide, making the search for new sources of water vital. As mine water could serve as a potential source, this study investigated the presence of microplastics in water from terminated deep mines in the largest coal basin in the Czech Republic, as well as in water from nearby shallow wells. The particles found were analyzed for size, polymer composition, color and morphology using the ImageJ tool, infrared spectroscopy with Fourier transform (FTIR) and an optical stereomicroscope with a digital camera. Microplastics were detected in all tested sites. Their range accounted for 2.5–17.5 items/L for mine water samples and 2.5–20 items/L for well samples, with fibers being the dominant type. The average width of particles from mine water and wells amounted to 58 µm; 71 µm, length to 655 µm; 501 µm and area to 22,067 µm2; 28,613 µm2, respectively. Blue color was prevalent, among materials, in both cases, plastic coated paper was found dominant to Polyethylene terephthalate (PET), Polyester (PES), Tetrafluoroethylene-perfluoro (Propyl Vinyl Ether) - Copolymer (TFE-PPVE), and polypropylene (PP). The research provides the first evidence of microplastics' presence in underground waters from deep mines and shallow wells in the same area. The data suggest that it is almost impossible to find underground water sources free of microplastic contamination. In this context, atmospheric contamination from mine ventilation and infiltration through terminated mines were identified as potential sources, while infiltration through soil and rock formations is unlikely given the geological composition. The results of this study can serve as a relevant basis for further research on microplastics in mine waters. Additionally, the conclusions can advance the development in remediation technologies of microplastics from deep underground waters and their implementation in practice, particularly in light of upcoming legislation.
气候变化正在给全球供水带来新的挑战，因此寻找新的水源至关重要。由于矿井水可能是一种潜在的来源，因此本研究调查了捷克共和国最大的煤盆地已终止的深矿井水中以及附近浅井水中微塑料的存在。使用 ImageJ 工具、带有傅里叶变换（FTIR）的红外光谱和带有数码相机的光学立体显微镜分析发现的颗粒的大小、聚合物组成、颜色和形态。在所有测试地点都检测到微塑料。矿井水样品的含量范围为2.5-17.5个项目/L，井水样品的含量范围为2.5-20个项目/L，其中纤维为主要类型。矿井水和井中颗粒的平均宽度为58 μm;71 μm，长度至 655 μm;501 μm，面积为 22,067 μm2;分别为28,613μm2。在两种材料中，蓝色普遍存在，在这两种情况下，塑料淋膜纸都以聚对苯二甲酸乙二醇酯（PET）、聚酯（PES）、四氟乙烯-全氟（丙基乙烯基醚）-共聚物（TFE-PPVE）和聚丙烯（PP）为主。该研究提供了微塑料存在于同一地区深矿和浅井地下水中的第一个证据。数据表明，几乎不可能找到没有微塑料污染的地下水源。在这方面，矿井通风造成的大气污染和通过已终止的矿井渗透被确定为潜在来源，而鉴于地质成分，通过土壤和岩层渗透的可能性不大。本研究结果可为进一步开展矿井水中微塑料研究提供参考。此外，这些结论可以推动来自深层地下水域的微塑料修复技术的发展及其在实践中的实施，特别是考虑到即将出台的立法。

Graphical abstract 图形摘要

Keywords 关键字

Microplastics

Mining

Groundwater

FTIR

Pollution

Characterization

微塑料

采矿

地下水

FTIR

污染

表征

1. Introduction 1. 引言

Due to climate change and population growth, exploring alternative water supply sources is necessary (da Silva and Blanco, 2023; Fonjong and Zama, 2023; Salehi, 2022). Among other water sources, mine water has already been utilized in some regions facing long-term water scarcity, mainly for domestic and irrigation purposes (Anjali et al., 2023; Chen et al., 2020; Gao et al., 2020). The Czech Republic is also experiencing prolonged droughts (Debiec, 2021) and has several mining localities with substantial water reserves (Wlosok and Vostarek, 2020).
由于气候变化和人口增长，探索替代供水来源是必要的（da Silva 和 Blanco，2023 年;Fonjong 和 Zama，2023 年;萨利希， 2022 年）。在其他水源中，矿井水已经在一些面临长期缺水的地区得到利用，主要用于家庭和灌溉目的（Anjali 等人，2023 年; Chen et al.， 2020;Gao 等人，2020 年）。捷克共和国也正在经历长期干旱（Debiec，2021 ），并有几个拥有大量水储量的矿区（Wlosok 和 Vostarek，2020 ）。

Understanding the composition of these mine waters is crucial, as it varies greatly depending on the location. It can range from clean water suitable for water infrastructure to contaminated mine waters that pose environmental challenges. The composition of mine water has been extensively discussed by various authors for decades (Arefieva et al., 2016; Banwart and Malmström, 2001; Kennedy et al., 2023; Lottermoser, 2010; Opitz et al., 2023; Tomiyama and Igarashi, 2022). Nevertheless, microplastics are still among the newly monitored contaminants in the aquatic environment (Pan et al., 2023; Reddy et al., 2021).
了解这些矿井水的成分至关重要，因为它会因位置的不同而有很大差异。它的范围从适合水利基础设施的清洁水到构成环境挑战的污染矿井水。几十年来，矿井水的成分一直被各种作者广泛讨论（Arefieva 等人，2016 年;Banwart 和 Malmström，2001 年; 肯尼迪等人，2023 年;Lottermoser，2010 年;Opitz 等人，2023 年;Tomiyama 和 Igarashi，2022 年）。尽管如此，微塑料仍然是水生环境中新监测的污染物之一（Pan 等人，2023 年;Reddy 等人，2021 年）。

Microplastics are defined as plastic particles smaller than 5 mm (Arthur et al., 2008; Halfar et al., 2021; Sparks et al., 2023) and can be categorized as primary or secondary depending on their origin (Galindo Montero et al., 2023; Lamichhane et al., 2023; Li et al., 2023; Nakat et al., 2023; Saeedi, 2023). Primary microplastics are intentionally manufactured for their specific properties. In contrast, secondary microplastics result from external influences, such as aging, degradation and fragmentation of plastic materials due to abiotic (mainly mechanical, thermal, photo and hydrolytic) and biotic (biological activities, e.g., bacteria, fungi, enzymes) processes (Bher et al., 2022; Dimassi et al., 2022; Rose et al., 2023; Zha et al., 2022). These mechanisms interact and lead to changes in the physicochemical properties of polymers. (He et al., 2023; Hu et al., 2023)
微塑料被定义为小于 5 毫米的塑料颗粒（Arthur 等人，2008 年;Halfar 等人，2021 年;Sparks 等人，2023 年）根据其来源，可分为主要或次要（Galindo Montero 等人，2023 年;Lamichhane 等人，2023 年; Li 等人，2023 年;Nakat 等人，2023 年;Saeedi，2023 年）。初级微塑料是有意制造的，具有特定的特性。相比之下，次生微塑料是由外部影响引起的，例如由于非生物（主要是机械、热、光解和水解）和生物（生物活动，例如细菌、真菌、酶）过程导致的塑料材料老化、降解和碎裂（Bher et al.， 2022;Dimassi 等人，2022 年;Rose 等人，2023 年;Zha 等人，2022）。这些机制相互作用并导致聚合物物理化学性质的变化。（He 等人，2023 年; 胡等，2023）

Microplastics occur in various forms, including fibers, fragments, flakes, films, spheres and more (Kurki-Fox et al., 2023; Rios Mendoza and Balcer, 2019). Their small size, large specific surface area, hydrophilicity and hydrophobicity can negatively impact the environment, as they act as carriers for other contaminants and can easily enter all components of the environment, including living organisms (K.l. et al., 2022).
微塑料以各种形式存在，包括纤维、碎片、薄片、薄膜、球体等（Kurki-Fox 等人，2023 年;里奥斯·门多萨（Rios Mendoza）和巴尔瑟（Balcer）， 2019年）。它们的体积小、比表面积大、亲水性和疏水性会对环境产生负面影响，因为它们充当其他污染物的载体，并且很容易进入环境的所有组成部分，包括生物体（K.l. 等人，2022）。

As a consequence, Directive (EU) 2020/2184 of the European Parliament and the Council introduced a mechanism for listing regulated substances, including microplastics. It is vital to consider the possibility that microplastics may be subject to restrictions in water designated for human consumption. Simultaneously, efforts are underway to develop ISO 24187, which outlines principles for analyzing microplastics in the environment.
因此，欧洲议会和理事会的指令（EU） 2020/2184 引入了一种列出受管制物质（包括微塑料）的机制。至关重要的是，要考虑到微塑料在指定供人类食用的水中可能受到限制的可能性。与此同时，正在努力制定 ISO 24187，它概述了分析环境中微塑料的原则。

Current research focuses on the occurrence of microplastics in different environments, including air, soil and water. As for water, surface and groundwater have been studied for MPs (Chia et al., 2022; Jeong et al., 2023; Kim et al., 2023; Mu et al., 2022; Ren et al., 2021; Sangkham et al., 2023; Shu et al., 2023; Singh and Bhagwat, 2022; Viaroli et al., 2022; Wu et al., 2022; Zhou et al., 2022). However, to our knowledge, there is currently no study addressing the issue of microplastics in mine waters. We hypothesize that despite the deep mine environment, MPs are present in deep mine water in terminated mines, likely as the result of the communication of the mine with the ground surface.
目前的研究主要集中在微塑料在不同环境中的发生，包括空气、土壤和水。至于水，已经为国会议员研究了地表水和地下水（Chia et al.， 2022;Jeong 等人，2023 年;Kim 等人，2023 年; Mu 等人，2022 年; 任等人，2021;Sangkham 等人，2023 年; Shu et al.， 2023;Singh 和 Bhagwat，2022 年;Viaroli 等人，2022 年; Wu et al.， 2022; 周等人，2022）。然而，据我们所知，目前还没有关于矿井水中微塑料问题的研究。我们推测，尽管矿井环境较深，但 MP 仍存在于已终止矿井的深矿水中，这可能是矿井与地表通信的结果。

Hence, the aim of this study is to detect the presence and identify microplastics in water from deep mines and nearby shallow wells to provide valuable data for further research on microplastics in groundwater. To determine the occurrence, shape, size, type of polymer and color of the microplastics we used Fourier transformation infrared spectroscopy.
因此，本研究旨在检测深矿和附近浅井水中的微塑料的存在并进行鉴定，为进一步研究地下水中的微塑料提供有价值的数据。为了确定微塑料的出现、形状、大小、聚合物类型和颜色，我们使用了傅里叶变换红外光谱。

The novelty of our study lies in being the first to prove the occurrence of microplastics in deep mine waters and providing characteristics of such pollutants so that effective methods to remove or mitigate their presence can be developed. Moreover, we detected microplastics in comparable quantities to samples taken from shallow wells in the same region, which is the largest coal basin in the Czech Republic. The presence of microplastics in mine water led to the detection of potential infiltration routes of microplastics into great depths. Identifying these routes together with characterizing the found microplastics is highly valuable for designing appropriate remediation technologies. Additionally, the study provides essential information on anthropogenic contamination in potential sources of groundwater, approximately 700 m below surface.
我们研究的新颖性在于首次证明了深矿水中微塑料的存在，并提供了此类污染物的特性，以便可以开发有效的方法来去除或减轻它们的存在。此外，我们检测到的微塑料数量与从同一地区的浅井中采集的样本相当，该地区是捷克共和国最大的煤炭盆地。矿井水中微塑料的存在导致人们发现了微塑料向深处的潜在渗透途径。确定这些途径并表征发现的微塑料对于设计适当的修复技术非常有价值。此外，该研究还提供了关于地表以下约700米的潜在地下水源中人为污染的重要信息。

2. Material and methods 2.材料与方法

2.1. Sampling sites and collection
2.1. 采样点和采集

Groundwater samples from mines and wells were collected at four locations in the Moravian-Silesian Region, Czechia, see Fig. 1. The region includes a densely populated agglomeration with approximately one million permanent residents. Moreover, coal mining and heavy industry have impacted the region a great deal.
在捷克摩拉维亚-西里西亚地区的四个地点采集了矿山和水井的地下水样本，见图 1。该地区包括一个人口稠密的集聚区，拥有约100万常住居民。此外，煤炭开采和重工业对该地区产生了很大影响。

Mine water was collected from two terminated black coal mines, Jeremenko Mine in Ostrava-Vítkovice and Žofie Mine in Orlová-Poruba. The water is continuously pumped from the mines and discharged into surface waters to prevent flooding and subsidence in the mining region. The pumped waters have a neutral pH and are saline (Arnošt Grmela, 2017; Arnošt et al., 2003). Additionally, water was sampled from two nearby shallow wells for comparison. These were metal wells without any plastic fittings and serve as sources of water for gardening and flushing in private houses located in Paskov.
矿井水是从两个已终止的黑煤矿收集的，即俄斯特拉发-维特科维采的Jeremenko煤矿和Orlová-Poruba的Žofie煤矿。水不断从矿井中抽出并排放到地表水中，以防止矿区发生洪水和沉降。泵送水的pH值为中性，是咸水（Arnošt Grmela，2017 年;Arnošt 等人，2003 年）。此外，还从附近的两口浅井中取样取水进行比较。这些是没有任何塑料配件的金属井，用作位于帕斯科夫的私人住宅的园艺和冲洗水源。

Water samples were collected following relevant international standards, EN 5667–11:2009 and EN ISO 5667–14:2006. The sampling took place between May 2022 and September 2022, over a 5-month period. Monthly, 400 mL of water were sampled – see the sampling sites in Fig. 1 and Table 1. A total of 20 samples were collected during this period, with 10 samples from shallow wells located 5 m below the surface (5 samples each from Well 1 and Well 2) and 10 samples of mine water (5 samples each from the Jeremenko pit and the Žofie pit). Trained workers collected mine water directly into brown glass bottles from the mine pit. Before sampling, the bottles were thoroughly washed with distilled and filtered water (A/C Glass Fiber, 1 μm, 47 mm, PALL Mexico), followed by a rinse with pure ethanol (VWR; 97%; CAS: 64–17–5). The bottles were sealed with glass stoppers and made ready for sampling. Due to the complex layout of the deep mine, the worker collected water one bottle at a time.
水样的采集遵循相关国际标准EN 5667-11：2009和EN ISO 5667-14：2006。抽样时间在 2022 年 5 月至 2022 年 9 月之间，为期 5 个月。每月取样 400 mL 水 – 参见图 1 和表 1 中的取样点。在此期间共收集了20个样品，其中10个样品来自地表以下5米的浅井（1号井和2号井各5个样品）和10个矿井水样品（Jeremenko坑和Žofie坑各5个样品）。训练有素的工人从矿坑中直接收集矿井水到棕色玻璃瓶中。取样前，用蒸馏水和过滤水（A/C 玻璃纤维，1 μm，47 mm，PALL Mexico）彻底清洗瓶子，然后用纯乙醇冲洗（VWR; 97%;CAS：64-17-5）。瓶子用玻璃塞密封，准备取样。由于深矿布局复杂，工人们一次收集一瓶水。

Table 1. Location of the selected sampling sites.
表 1.选定采样点的位置。

Location 位置	GPS coordinates GPS坐标	Elevation (MASL) 高程（MASL）	Sampling depth (m) 采样深度（m）
Jeremenko mine Jeremenko矿	49°48′18.3″N 18°16′12.8″E 49°48 ′ 18.3 “N 18°16 ′ 12.8”东	232.3	622
Žofie mine	49°50′45.4″N 18°25′15.4″E 49°50 ′ 45.4“N 18°25 ′ 15.4”东	233.6	709
Well 1 井 1	49°43′56.6″N 18°17′53.4″E 49°43′56.6“N 18°17′53.4”东	255.6	5
Well 2 井 2	49°43′41.6″N 18°17′58.3″E 49°43′41.6“N 18°17′58.3”东	257.8	5

Shallow well groundwater samples were obtained using identically prepared containers. Water was sampled directly from the wells using a metal bucket and poured into the prepared sampling bottle. Next, samples were immediately transported to the lab in a cooler for analysis and thus no sample stabilization was required.
浅井地下水样品是使用相同制备的容器获得的。使用金属桶直接从井中取样水，然后倒入准备好的取样瓶中。接下来，样品立即在冷却器中运送到实验室进行分析，因此不需要样品稳定。

2.2. Quality control 2.2. 质量控制

Precautions were taken to prevent contamination from plastic materials during the sampling process and while using sampling tools. We followed the guidelines outlined in the EN ISO 5667–14:2006 standard for sample collection, which focuses on reducing errors during sampling. All staff involved in handling the samples received proper training and were familiar with the sampling protocols.
在取样过程中和使用取样工具时，采取了预防措施以防止塑料材料的污染。我们遵循 EN ISO 5667–14：2006 标准中概述的样本采集指南，该标准侧重于减少采样过程中的错误。所有参与处理样本的工作人员都接受了适当的培训，并熟悉采样规程。

Mine water samples were collected using amber glass storage bottles, which were pre-washed with pure ethanol (VWR; 97%; CAS: 64–17–5) and wrapped in aluminum foil. The operator wore disposable latex gloves and cotton clothing during the process. Mine water samples were directly obtained from the pumped pits and shallow groundwater samples were collected using steel buckets from the wells and transferred to amber glass storage bottles. The samples were immediately sealed and stored at 4 °C for future analysis. In the laboratory, before filtration, all surfaces were rinsed with pure ethanol (VWR; 97%; CAS: 64–17–5) and cleaned with a cotton towel. Only one person was present in the lab and the air conditioner was turned off to minimize potential contamination. All laboratory glassware was rinsed with clean ethanol (VWR; 97%; CAS: 64–17–5) and covered with aluminum foil. Throughout the process, cotton coats and disposable nitrile gloves were used. During filtration, a blank test was conducted by placing dry filter paper (A/C Glass Fiber, 1 μm, 47 mm, PALL Mexico) in an open Petri dish alongside the single sample filtration to assess air contamination.
矿井水样品是使用琥珀色玻璃储存瓶收集的，这些储存瓶用纯乙醇（VWR; 97%;CAS：64-17-5）并用铝箔包裹。操作人员在此过程中佩戴一次性乳胶手套和棉质衣物。矿井水样直接从抽水坑中获取，浅层地下水样使用井中的钢桶收集，然后转移到琥珀色玻璃储存瓶中。将样品立即密封并储存在4°C以备将来分析。在实验室中，在过滤之前，所有表面都用纯乙醇（VWR;CAS：64-17-5），并用棉毛巾清洁。实验室里只有一个人在场，空调被关闭，以尽量减少可能的污染。所有实验室玻璃器皿均用干净的乙醇（VWR;97%;CAS：64-17-5）并用铝箔覆盖。在整个过程中，使用了棉质外套和一次性丁腈手套。在过滤过程中，通过将干燥滤纸（A/C 玻璃纤维，1 μm，47 mm，PALL Mexico）与单个样品过滤一起放入开放式培养皿中进行空白测试，以评估空气污染。

2.3. Microplastic extraction and identification
2.3. 微塑料的提取和鉴定

Water samples, each with a volume of 400 ml, were transferred to clean glass beakers and prepared for filtration. Due to limited filter capacity, each sample was divided into two beakers, with a volume of 200 ml each. In total, 20 samples (40 filters) were analyzed: 10 samples (20 filters) of groundwater from two wells (Well 1, Well 2) and 10 samples (20 filters) of mine water from two mines (Žofie Mine, Jeremenko Mine). All samples were filtered using vacuum filtration through glass fiber filters (A/C Glass Fiber, 1 μm, 47 mm, PALL Mexico). After filtration, the glass fiber filters were carefully transferred to glass Petri dishes using tweezers. Subsequently, the Petri dishes were placed in a desiccator at room temperature for 48 h to dry.
将每个体积为400ml的水样转移到干净的玻璃烧杯中并准备过滤。由于过滤容量有限，每个样品被分成两个烧杯，每个烧杯的体积为 200 ml。总共分析了 20 个样品（40 个过滤器）：来自两口井（1 号井、2 号井）的 10 个样品（20 个过滤器）和来自两个矿井（Žofie 矿、Jeremenko 矿）的 10 个样品（20 个过滤器）。所有样品均通过玻璃纤维滤光片（A/C 玻璃纤维，1 μm，47 mm，PALL Mexico）使用真空过滤进行过滤。过滤后，用镊子小心地将玻璃纤维过滤器转移到玻璃培养皿中。随后，将培养皿置于室温干燥器中48小时以干燥。

After filtration and drying, the samples underwent a preliminary visual analysis using a stereomicroscope (SZX10, Olympus, Japan) with a 60x magnification digital camera (Canon EOS 5000, Canon, Japan). Suspected particles were selected and their properties (color, particle type, size) were assessed for each sample. The total number of suspected particles was optically quantified and photographed.
过滤和干燥后，使用体视显微镜（SZX10，奥林巴斯，日本）和60倍放大数码相机（佳能EOS 5000，佳能，日本）对样品进行了初步的视觉分析。选择可疑颗粒，并评估每个样品的特性（颜色、颗粒类型、大小）。对可疑颗粒的总数进行光学量化和拍照。

The particles were analyzed using ImageJ (Schneider et al., 2012, p. 25) to determine their size, including length, width and area. The threshold function was used to modify the particles and their maximum length, maximum width and total area were measured by outlining the entire particle.
使用 ImageJ（Schneider 等人，2012 年，第 25 页）分析颗粒以确定其大小，包括长度、宽度和面积。利用阈值函数对粒子进行修饰，通过勾勒出整个粒子的轮廓来测量粒子的最大长度、最大宽度和总面积。

Then, Fourier transform infrared spectroscopy (μ-FTIR) was performed on a Nicolet iN10 instrument (Thermo Fisher Scientific, USA) with an MCT detector cooled by liquid nitrogen. Each sample, having 2 filters, allowed selecting a maximum of 10 particles for μ-FTIR analysis from each filter. Ten particles were manually chosen from each filter using tweezers and placed on a glass slide for analysis under a microscope. The particles were measured in reflection mode with 64 scans in high quality, covering the spectral range of 400–4000 cm⁻¹. Under the microscope, the shape and color of the particles were primarily determined. The obtained vibrational spectra and the measured particles were compared with 5 spectral libraries of plastic materials and plastic additives. A threshold of over 60% match with the spectra in the library was set to confirm the polymer composition of particles. For matches within the range 50%−70% manual analysis was conducted to compare the characteristic peaks of the identified plastic material. Finally, the experimental data obtained were evaluated and subjected to statistical processing.
然后，在Nicolet iN10仪器（Thermo Fisher Scientific，USA）上进行傅里叶变换红外光谱（μ-FTIR），该仪器带有由液氮冷却的MCT检测器。每个样品有 2 个滤光片，允许从每个滤光片中选择最多 10 个颗粒进行 μ-FTIR 分析。使用镊子从每个过滤器中手动选择10个颗粒，然后放在载玻片上，在显微镜下进行分析。这些颗粒在反射模式下进行了64次高质量扫描，覆盖了400-4000 cm⁻¹的光谱范围。在显微镜下，主要确定了颗粒的形状和颜色。将得到的振动光谱和被测颗粒与塑料材料和塑料添加剂的5个光谱库进行了比较。设定了与谱库中光谱匹配度超过60%的阈值，以确认颗粒的聚合物组成。对于50%−70%范围内的匹配，进行了手动分析，以比较鉴定出的塑料材料的特征峰。最后，对得到的实验数据进行评估和统计处理。

The technique was selected based on methods reported in the available literature (Table 3) and due to the absence of a standardized methodology for analyzing microplastics in groundwater. It is important to mention that ISO 24187 - "Principles for the analysis of microplastics present in the environment" is currently under preparation. Once adopted, this could help standardize both the data obtained and the methodologies used.
该技术的选择基于现有文献中报告的方法（表 3），并且由于缺乏分析地下水中微塑料的标准化方法。值得一提的是，ISO 24187 - “环境中存在的微塑料分析原则”目前正在制定中。一旦获得通过，这将有助于使所获得的数据和所使用的方法标准化。

2.4. Statistical analysis
2.4. 统计分析

Factor Analysis of Mixed Data (FAMD) was employed to examine overall trends in the data, allowing a simultaneous assessment of qualitative and quantitative active variables. The variables used for constructing the model were particle shape, particle color and the number of estimated plastic particles. Additionally, variables like sample (two wells and two mines), site (mine X well) and sampling month were projected to the model.
混合数据因子分析（FAMD）用于检查数据的总体趋势，从而可以同时评估定性和定量的活动变量。用于构建模型的变量是颗粒形状、颗粒颜色和估计的塑料颗粒数量。此外，样本（两口井和两口矿井）、地点（X井）和采样月份等变量也被预测到模型中。

To assess the dependence of the number of estimated plastic particles on the sample, site and sampling month, as well as the combination of the sampling month and sample or site, one-way analysis of variance (ANOVA) was applied. The analyses and visualizations were conducted using the R environment (R Core Team, 2021), specifically utilizing the FactoMineR (Lê et al., 2008) and factoextra packages (Kassambara and Mundt, 2020).
为了评估估计的塑料颗粒数量对样品、地点和采样月份的依赖性，以及采样月份和样品或地点的组合，应用了单因素方差分析（ANOVA）。分析和可视化是使用 R 环境（R Core Team，2021 年）进行的，特别是利用 FactoMineR（Lê 等人，2008 年）和 factoextra 包（Kassambara 和 Mundt，2020 年）。

3. Results and discussion
3. 结果与讨论

3.1. Microplastics abundance
3.1. 微塑料的丰度

Microplastic presence was verified in all four sampling locations of mine water and shallow groundwater wells. Utilizing the μ-FTIR method, 302 out of 445 suspected particles were examined. Among these analyzed particles, 80 were positively identified as plastic or plastic additives, comprising approximately 27% of the total. The total microplastic abundance across all samples ranged from 2.5 to 20 items/L. To facilitate a more comprehensive understanding of the sampling distribution and the number of identified items, we have provided Table 2 below.
在矿井水和浅层地下水井的所有四个采样点都验证了微塑料的存在。利用 μ-FTIR 方法，检查了 445 个可疑颗粒中的 302 个。在这些分析的颗粒中，有80个被肯定为塑料或塑料添加剂，约占总数的27%。所有样品的总微塑料丰度范围为 2.5 至 20 件/升。为了更全面地了解抽样分布和已识别项目的数量，我们在下文提供了表 2。

Table 2. Number of items in samples (sample volume 400 ml).
表 2.样品中的项目数（样品体积 400 ml）。

Sample 样本	Date (day.month.year) 日期（日、月、年）	Selected items from stereomicroscope 体视显微镜的精选项目	Measured items by FTIR FTIR测量项目	Confirmed plastic/additives items by FTIR FTIR确认的塑料/添加剂项目
Well 1 井 1	08.05.2022	12	12	8
Well 2 井 2	08.05.2022	10	10	3
Well 1 井 1	15.06.2022	32	20	1
Well 2 井 2	15.06.2022	32	20	6
Well 1 井 1	22.07.2022	20	15	5
Well 2 井 2	22.07.2022	22	16	3
Well 1 井 1	05.08.2022	22	16	6
Well 2 井 2	05.08.2022	19	15	3
Well 1 井 1	12.09.2022	14	14	4
Well 2 井 2	12.09.2022	11	11	6
Jeremenko 杰列缅科	07.05.2022	19	18	6
Žofie 索菲亚	07.05.2022	16	16	7
Jeremenko 杰列缅科	15.06.2022	33	20	5
Žofie 索菲亚	15.06.2022	39	20	3
Jeremenko 杰列缅科	16.07.2022	30	17	4
Žofie 索菲亚	16.07.2022	54	20	4
Jeremenko 杰列缅科	23.08.2022	14	14	2
Žofie 索菲亚	23.08.2022	7	7	2
Jeremenko 杰列缅科	21.09.2022	7	7	1
Žofie 索菲亚	21.09.2022	32	14	1

In the well samples, there were 194 suspected particles, of which 149 were analyzed using μ-FTIR. Among these particles, 45 were confirmed as plastic or plastic additives, accounting for 30% of the total. The microplastic abundance was found to be 2.5–20 items/L.
在井样中，有194个疑似颗粒，其中149个使用μ-FTIR进行分析。在这些颗粒中，有45种被确认为塑料或塑料添加剂，占总数的30%。发现微塑料丰度为 2.5-20 件/L。

The mine water samples exhibited a lower microplastics load. Out of 251 suspected particles, 153 were analyzed using μ-FTIR, but only 35 were confirmed as plastic or plastic additives, making up 23% of the total. The microplastic abundance ranged from 2.5 to 17.5 items/L.
矿井水样品的微塑料含量较低。在 251 个疑似颗粒中，有 153 个使用 μ-FTIR 进行了分析，但只有 35 个被确认为塑料或塑料添加剂，占总数的 23%。微塑料丰度范围为2.5至17.5件/升。

The highest abundance of microplastics, 20 items/L, was found in the first well sample collected on 08.05.2022. The second-highest concentration of microplastics, 17.5 items/L, was observed in the Žofie Mine sample, collected on 07.05.2022. Microplastics found in well water likely come from anthropogenic sources. They are able to infiltrate through the soil to a depth, in our case of 5 m, into the groundwater (Khant and Kim, 2022). For further comparison, Table 3 displays studies that have investigated microplastics in groundwater.
在 2022 年 5 月 8 日收集的第一个井样品中发现微塑料丰度最高，为 20 件/升。在 2022 年 5 月 7 日采集的 Žofie 矿样品中观察到第二高的微塑料浓度，为 17.5 件/升。在井水中发现的微塑料可能来自人为来源。它们能够通过土壤渗透到深度，在我们的例子中为 5 m，进入地下水（Khant 和 Kim，2022 年）。为了进一步比较，表 3显示了对地下水中微塑料进行调查的研究。

Table 3. Studies focusing on microplastics in groundwater.
表 3.重点关注地下水中的微塑料研究。

Reference 参考	Sampling location 采样地点	Water type 水类型	Analytic technique 分析技术	Abundance 丰富	Most common polymer composition of MPs MPs最常见的聚合物组成
(Jeong et al., 2023) （Jeong 等人，2023 年）	Eastern Korea 朝鲜东部	groundwater 地下水	μ-FT-IR μ-傅立叶红外光谱	7–513 particles/500L 7–513 颗粒/500L	PP, PE, PET PP、PE、PET系列
(Samandra et al., 2022) （Samandra 等人，2022 年）	Victoria, Australia 维多利亚，澳大利亚	groundwater 地下水	LDIR	16–97 particles/L 16–97 颗粒/L	PE, PP, PS, PVC PE、PP、PS、PVC
(Shu et al., 2023) （Shu et al.， 2023）	Southwest China 西南地区	groundwater 地下水	Raman/FTIR 拉曼/傅里叶变换红外光谱	0–4 items/L 0–4 项/L	PP, PS, PET PP、PS、PET系列
(Esfandiari et al., 2022) （Esfandiari 等人，2022 年）	Southwest Iran 伊朗西南部	groundwater 地下水	Binocular microscope+heated probe 双目显微镜+加热探针	0.1–1.3 MP/L 0.1–1.3 英里/升	PS, PE, PET PS、PE、PET系列
(Balestra et al., 2023) （Balestra 等人，2023 年）	Italy 意大利	water from cave 洞穴中的水	μ-FT-IR μ-傅立叶红外光谱	12–54 items/L 12–54 件/升	PE, PVC, PES PE、PVC、PES系列
(Valsan et al., 2023) （Valsan 等人，2023 年）	southwest India 印度西南部	groundwater 地下水	FTIR-ATR	0.15 (±0.1) MPs/L 0.15 （±0.1） MPs/L	PES, LDPE 聚醚醚酮、低密度聚乙烯
(Lapworth and Shockley, 2022) （拉普沃思和肖克利， 2022 年）	UK	groundwater 地下水	FTIR	0.4 MPs/L 0.4 英里/升	PP, PE, PS, PC, acrylate PP、PE、PS、PC、丙烯酸酯
(Kim et al., 2023) （Kim 等人，2023 年）	Jeju volcanic Island, Korea 韩国济州火山岛	groundwater 地下水	μ-FT-IR μ-傅立叶红外光谱	0.006 to 0.192 particles/L 0.006 至 0.192 颗粒/升	PP, PE PP、PE

Legend: PP = polypropylene, PE = polyethylene, PET = polyethylene terephthalate, PS = polystyrene, PVC = Polyvinyl chloride, PES = polyethersulfone, LDPE = low-density polyethylene, PC = polycarbonate.
图例：PP = 聚丙烯，PE = 聚乙烯，PET = 聚对苯二甲酸乙二醇酯，PS = 聚苯乙烯，PVC = 聚氯乙烯，PES = 聚醚砜，LDPE = 低密度聚乙烯，PC = 聚碳酸酯。

The comparison of the abundance of microplastics in mine waters with other regions, cities, or states is currently not feasible due to the lack of similar studies. As for the origin of MPS, in the Czech Republic the presence of microplastics at significant depths below the surface might be linked to mine ventilation. During mine operation, forced air flow with fans creates a negative pressure, allowing fresh air distribution to all parts of the mine. Consequently, microplastics from the surface air could have accumulated in mine shafts to be later released into mine waters while flooding the mine upon termination. (Personal communication, 10/12/2022) Atmospheric transport as a potential cause of microplastic occurrence in underground spaces is supported by findings from Baraza in a cave environment (Baraza and Hasenmueller, 2023). However, other pathways cannot be ruled out. Surface water influx during heavy precipitation events might carry anthropogenic particles, contributing to the microplastic presence. The extensive surface area of deep mines and insufficient mapping of potential underground pathways add to the uncertainty. Infiltration of polymer particles through soil and rock environments is another possible pathway, but this process is influenced by various factors like particle size, polymer composition, hydrophobicity and specific soil types (Guo et al., 2022). Fig. 2 shows a negative Pearson correlation between precipitation and microplastics in mine water (r = −0.61 for Jeremenko and r = −0.68 for Žofie). However, the correlation was not found to be significant. There was also a lower negative correlation to the groundwater samples from wells (r = −0.24 and t = −0.13), which suggests that precipitation might not be the sole factor affecting microplastic abundance in the selected area.
由于缺乏类似的研究，目前无法将矿井水中微塑料的丰度与其他地区、城市或州进行比较。至于MPS的起源，在捷克共和国，地表以下大量深度的微塑料的存在可能与矿井通风有关。在矿井作业期间，通过风扇的强制气流会产生负压，使新鲜空气能够分布到矿井的所有部分。因此，地表空气中的微塑料可能会在矿井中积聚，然后在矿井结束时释放到矿井中，同时淹没矿井。（个人来文，2022年10月12日）大气迁移是地下空间中微塑料发生的潜在原因，得到了 Baraza 在洞穴环境中的发现的支持（Baraza 和 Hasenmueller，2023 年）。但是，不能排除其他途径。在强降水事件期间，地表水的流入可能会携带人为颗粒物，从而导致微塑料的存在。深部矿山的广阔表面积和潜在地下通道的测绘不足增加了不确定性。聚合物颗粒通过土壤和岩石环境渗透是另一种可能的途径，但这一过程受到颗粒大小、聚合物组成、疏水性和特定土壤类型等各种因素的影响（Guo et al.， 2022）。图 2显示了矿井水中降水与微塑料之间的负Pearson相关性（Jeremenko的r = -0.61，Žofie的r = -0.68）。然而，没有发现相关性显着。与井中地下水样品的负相关也较低（r = −0.24和t = −0.13），这表明降水可能不是影响所选地区微塑料丰度的唯一因素。

3.2. Microplastics characterization - morphology, color and size
3.2. 微塑料表征 - 形态、颜色和大小

The samples contained particles with variations in shape, color and size. Four distinct shapes were identified: fibers, fragments, particles and film. Among all the samples, fibers were the most prevalent, as depicted in Fig. 3; S. 1 and S. 2. This dominance of fibers aligns with findings from previous studies examining groundwater microplastics (Baraza and Hasenmueller, 2023; Esfandiari et al., 2022; Fajaruddin Natsir et al., 2021; Shu et al., 2023; Wu et al., 2022). However, there are studies that emphasize the prevalence of microplastic fragments in groundwater (Cha et al., 2023; Jeong et al., 2023; Kim et al., 2023). This suggests that the type of particles found might be related to the specific utilization of the location.
这些样品含有形状、颜色和大小各不相同的颗粒。鉴定出四种不同的形状：纤维、碎片、颗粒和薄膜。在所有样品中，纤维是最普遍的，如图3所示 ;第1条及第2条。纤维的这种主导地位与之前检查地下水微塑料的研究结果一致（Baraza 和 Hasenmueller，2023 年;Esfandiari 等人，2022 年;Fajaruddin Natsir 等人，2021 年; Shu et al.， 2023; Wu 等人，2022 年）。然而，有研究强调地下水中微塑料碎片的普遍存在（Cha et al.， 2023;Jeong 等人，2023 年;Kim 等人，2023 年）。这表明发现的颗粒类型可能与该位置的特定利用有关。

In contrast, the presence of films in our samples was either infrequent or nonexistent. The irregular shape of fragments and films indicates that they likely originated from the breakdown of larger plastic objects like plastic bags, bottles, or foils. On the other hand, fibers exhibit a regular shape and varying lengths, suggesting their possible sources as synthetic clothing, upholstery, or the breakdown of larger plastic pieces (Chaudhari and Samnani, 2022).
相比之下，我们的样本中薄膜的存在要么很少见，要么根本不存在。碎片和薄膜的不规则形状表明它们可能起源于塑料袋、瓶子或箔纸等较大塑料物体的分解。另一方面，纤维表现出规则的形状和不同的长度，这表明它们可能的来源是合成服装、室内装潢或较大塑料片的分解（Chaudhari 和 Samnani，2022 年）。

Fig. 3 illustrates the distribution of different microplastic morphologies in the samples. Out of the 80 confirmed microplastics, fibers accounted for the largest portion at 59%, followed by particles (20%), fragments (19%) and films (2%). In both mine and well samples, fibers were the most abundant, making up 49% and 67% respectively. However, films were only observed in the mine water samples.
图 3显示了样品中不同微塑料形态的分布。在80种已确认的微塑料中，纤维占最大比例，为59%，其次是颗粒（20%）、碎片（19%）和薄膜（2%）。在矿山和油井样品中，纤维含量最高，分别占49%和67%。然而，仅在矿井水样中观察到薄膜。

Considerable color variability was observed, with a total of 11 distinct colors identified. Based on the data in Fig. 4, the most prevalent colors were as follows: blue (27%), gray (24%), brown (18%), transparent (10%), black (6%), white (4%), pink (4%), red (3%), beige (2%), orange (1%) and silver (1%). Notably, the dominance of the blue color aligns with findings from previous studies by Wu et al. and Zhang et al. However, our study observed a relatively lower prevalence of the transparent color, contrasting with the higher representation reported in their respective studies (Wu et al., 2022; Zhang et al., 2019). In contrast, K et al. reported blue as the second least abundant color (K et al., 2021). The chi-squared test revealed no significant differences in color distribution between the mine water and well samples (p = 0.17).
观察到相当大的颜色变异性，总共识别出 11 种不同的颜色。根据图 4中的数据，最流行的颜色如下：蓝色（27%）、灰色（24%）、棕色（18%）、透明（10%）、黑色（6%）、白色（4%）、粉红色（4%）、红色（3%）、米色（2%）、橙色（1%）和银色（1%）。值得注意的是，蓝色的主导地位与Wu等人和Zhang等人先前的研究结果一致。然而，我们的研究观察到透明色的患病率相对较低，这与他们各自研究中报告的较高代表性形成鲜明对比（Wu 等人，2022 年;Zhang 等人，2019 年）。相比之下，K 等人报告说蓝色是第二不丰富的颜色（K 等人，2021 年）。卡方检验结果显示，矿井水和油井样品的颜色分布无显著差异（p = 0.17）。

In well samples, the most prevalent colors were gray (33%), blue (23%) and brown (20%), while no white, silver, red, or orange particles were observed in the selected samples (S. 3a). In the first well, the largest share of particles was gray (29.5%) and brown (29.5%), while the second well had the highest proportion of gray particles at 38%.
在油井样品中，最普遍的颜色是灰色（33%）、蓝色（23%）和棕色（20%），而在选定的样品中没有观察到白色、银色、红色或橙色颗粒（S.3a）。在第一口井中，灰色（29.5%）和棕色（29.5%）的颗粒所占比例最大，而第二口井的灰色颗粒所占比例最高，为38%。

In mine samples, blue particles were the most abundant at 34%, followed by brown (14%) and gray (11%) (S. 3b). The Jeremko mine had the highest proportion of blue particles at 39%, while in Žofie, the color blue appeared most often at 29%.
在矿山样本中，蓝色颗粒的含量最高，为34%，其次是棕色（14%）和灰色（11%）（S.3b）。Jeremko矿的蓝色颗粒比例最高，为39%，而在Žofie，蓝色出现频率最高，为29%。

The analysis of microplastics in the samples focused on their size, specifically length, width and area. The results were categorized into six size groups: 0–50 µm, 50–100 µm, 100–250 µm, 250–500 µm, 500–1000 µm, and above 1000 µm. Additionally, for a more detailed view of the microplastic surface area, four distinct categories were established: 1000–10,000 µm², 10,000–15,000 µm², 15,000–25,000 µm² and 25,000–600,000 µm².
样品中微塑料的分析主要集中在它们的大小上，特别是长度、宽度和面积。结果分为6个尺寸组：0-50 μm、50-100 μm、100-250 μm、250-500 μm、500-1000 μm和1000 μm以上。此外，为了更详细地观察微塑料表面积，建立了四个不同的类别：1000-10,000 μm²、10,000-15,000 μm²、15,000-25,000 μm² 和 25,000-600,000 μm²。

Fig. 5 presents the findings related to microplastics discovered in mine waters. The particles in mine waters exhibited lengths ranging 58–3822 µm, widths spanning 5.7–226.6 µm and areas covering values between 1088 and 116,579 µm². In contrast, microplastics obtained from wells showed different ranges. The length varied 104.7–1798 µm, the width 7.6–692.6 µm and the area was recorded within the interval of 2647.6–520,626 µm².
图 5显示了与在矿井水中发现的微塑料相关的发现。矿井水中的颗粒长度范围为58-3822 μm，宽度范围为5.7-226.6 μm，面积范围在1088-116,579 μm之间²。相比之下，从孔中获得的微塑料显示出不同的范围。长度变化为104.7-1798 μm，宽度为7.6-692.6 μm，面积记录在2647.6-520,626 μm²的区间内。

The most prevalent microplastics were those with a width ranging from 0 to 50 µm, accounting for 50 pieces (62.5%) of all 80 identified microplastics. This group of microplastics prevailed and was the dominant component in the analyzed samples.
最普遍的微塑料是宽度范围为0至50μm的微塑料，占所有80种已确定的微塑料的50块（62.5%）。这组微塑料占主导地位，是分析样品中的主要成分。

Regarding the length of microplastics, the most numerous were those in the range of 100–250 µm and 250–500 µm, representing 24 items (30%) for each category. Together, these two categories constituted a significant portion of the microplastics (60%) that we recorded. Table 4 presents a comparison between the morphology, abundance and size of microplastics found in both mine and well water samples.
关于微塑料的长度，数量最多的是100-250μm和250-500μm之间的微塑料，代表每个类别24个项目（30%）。这两类微塑料共同构成了我们记录的微塑料（60%）的很大一部分。表 4比较了在矿井和井水样本中发现的微塑料的形态、丰度和大小。

Table 4. Comparison of microplastics morphology, abundance and size of samples from mine water and wells.
表 4.矿井和井中微塑料形态、丰度和样品大小的比较。

Type of groundwater 地下水的种类	Morphology 形态学	Abundance 丰富	Size characterization 尺寸表征	Average size 平均大小	Min. 最小值	Max. 麦克斯。
Mine water 矿井水	fiber 纤维	21	length µm 长度 μm	1156.2	245	3822
			width µm 宽度 μm	31.5	7.6	155
			area µm² 面积^{μm 2}	32,899.8	5072	116,579
	particle 粒子	10	length µm 长度 μm	132.3	66.3	217.5
			width µm 宽度 μm	86.3	45.1	136
			area µm² 面积^{μm 2}	9379.2	1922.4	20,042
	fragment 片段	9	length µm 长度 μm	295.6	58	670
			width µm 宽度 μm	76	5.7	226.6
			area µm² 面积^{μm 2}	16,194.7	1088	63,919
	film 影片	2	length µm 长度 μm	96.5	91	102
			width µm 宽度 μm	91	45	137
			area µm² 面积^{μm 2}	7165	3327	11,003
Well 井	fiber 纤维	36	length µm 长度 μm	618.8	138.5	1798
			width µm 宽度 μm	24.5	7.6	102.9
			area µm² 面积^{μm 2}	14,025.5	2647.6	64,411
	particle 粒子	10	length µm 长度 μm	176.3	104.7	357.3
			width µm 宽度 μm	146.8	61.5	429.5
			area µm² 面积^{μm 2}	26,959.6	5393	126,770
	fragment 片段	7	length µm 长度 μm	367.6	135.8	860.5
			width µm 宽度 μm	184.4	37.5	692.6
			area µm² 面积^{μm 2}	93,020.2	8181.7	520,626

Our results were compared with Balesta et al.'s study, which reported 78% of microplastics within the 100–990 µm length range (Balestra et al., 2023). In our research, we observed a similar outcome, with 88.75% of microplastics falling within this specific length range.
我们的结果与 Balesta 等人的研究进行了比较，该研究报告了 78% 的微塑料在 100-990 μm 长度范围内（Balestra 等人，2023 年）。在我们的研究中，我们观察到了类似的结果，88.75%的微塑料属于这个特定的长度范围。

3.3. Microplastics characterization - polymer composition
3.3. 微塑料表征 - 聚合物成分

Thirty-one types of microplastics were measured and identified in all samples.
在所有样品中测量和鉴定了 31 种类型的微塑料。

In the analysis of water from wells, plastic-coated paper was the most abundant plastic material type, with 15 identified particles. Other substances found included Polyethylene terephthalate (PET), Polyester (PES) and Tetrafluoroethylene-perfluoro (Propyl Vinyl Ether) Copolymer (TFE-PPVE), each accounting for 7% of the total sample volume.
在对井水的分析中，塑料淋膜纸是最丰富的塑料材料类型，有15个已确定的颗粒。发现的其他物质包括聚对苯二甲酸乙二醇酯（PET）、聚酯（PES）和四氟乙烯-全氟（丙基乙烯基醚）共聚物（TFE-PPVE），每种物质均占总样品体积的 7%。

In the analysis of water from mines, plastic-coated paper accounted for 27% of the total particles, while polyester (PES), polyethylene terephthalate (PET) and polypropylene (PP) made up 18%, 12% and 9% of the total volume, respectively (see Fig. 6).
在矿井水的分析中，塑料涂层纸占总颗粒量的27%，而聚酯（PES）、聚对苯二甲酸乙二醇酯（PET）和聚丙烯（PP）分别占总体积的18%、12%和9%（见图 6）。

The identified particles in the samples can originate from various sources, including packaging, industrial waste, or other waterborne contaminants. The high prevalence of plastic-coated paper in all samples strongly suggests its origin in the packaging industry. Chen et al. provide an example, mentioning the widespread use of disposable paper cups and PE-coated packaging as potential sources (Chen et al., 2023).
样品中识别出的颗粒可能来自各种来源，包括包装、工业废物或其他水性污染物。塑料淋布纸在所有样品中的高含量强烈表明其起源于包装行业。Chen et al. 提供了一个例子，提到一次性纸杯和 PE 涂层包装的广泛使用是潜在来源（Chen et al.， 2023）。

3.4. Statistical analysis
3.4. 统计分析

Fig. 7 visualizes the results of FAMD for the variables. The first two dimensions explain a similar amount of variability in the data. The only quantitative variable, the number of estimated plastic particles, showed a strong positive correlation with the first dimension (r = 0.81, p = 0.4 . 10⁻²²). Particle color, sampling month and particle shape were also significantly associated with this dimension (R2 = 0.68, R2 = 0.38 and R2 = 0.28, respectively). The second dimension was associated with particle shape, color and sample (R2 = 0.75, R2 = 0.78 and R2 = 0.11, respectively). The significant association of both sampling months and sampling site is noteworthy since these variables were only projected to the model and not used for its construction. However, the sampling site was not linked to any of the dimensions, as evident when the individuals (i.e., individual particles) were plotted on the first factor plane based on their sampling site (Fig. 8).
图 7 可视化了变量的 FAMD 结果。前两个维度解释了数据中相似的可变性量。唯一的定量变量，即估计的塑性颗粒数量，与第一维（r = 0.81，p = 0.4 . 10⁻²²）呈强正相关。颗粒颜色、采样月份和颗粒形状也与该维度显著相关（R2 = 0.68、R2 = 0.38和R2 = 0.28）。第二个维度与颗粒形状、颜色和样品相关（R2 = 0.75、R2 = 0.78 和 R2 = 0.11）。值得注意的是，采样月份和采样地点之间的显著关联是值得注意的，因为这些变量只是投影到模型中，并未用于模型的构建。然而，采样点与任何维度都没有关联，当个体（即单个粒子）根据其采样点绘制在第一因子平面上时，这一点很明显（图 8）。

The results of One-way Analysis of variance corroborated the aforementioned phenomenon. There was no significant difference found in the number of estimated plastic particles determined in mine waters and wells (Fig. 9a). On the other hand, there was a significant difference (p = 0.021) between the samples (Fig. 9b). While the samples from wells were rather similar in the number of plastic particles estimated, samples from mines were found to be much more variable – both between the samples and in the samples. In addition, the sampling month was found to have a significant effect (p = 0.11 . 10⁻⁵) on the number of estimated plastic particles as well as the combination of the sampling month and the sample (p = 0.2 . 10⁻¹⁵, two-way ANOVA).
单因素方差分析结果证实了上述现象。在矿井和油井中测定的估计塑料颗粒数量没有发现显着差异（图 9a）。另一方面，样品之间存在显着差异（p = 0.021）（图 9b）。虽然来自井的样品在估计的塑料颗粒数量上相当相似，但发现来自矿山的样品的可变性要大得多——无论是在样品之间还是在样品中。此外，还发现采样月份对估计的塑料颗粒数量以及采样月份和样品的组合具有显着影响（p = 0.2 . 10⁻¹⁵，双向方差分析）。

3.5. Remediation strategies in water sources
3.5. 水源地修复策略

Concerningly, microplastics are found in significant quantities both in shallow groundwater and deep water. As mentioned previously, it is highly probable that legislation will restrict the presence of microplastics in water sources in the future. Consequently, addressing the issue of their removal from water bodies becomes essential. This calls for supplementing the existing water treatment technologies with processes specifically designed to eliminate microplastics.
令人担忧的是，在浅层地下水和深层水中都发现了大量的微塑料。如前所述，未来立法极有可能限制水源中微塑料的存在。因此，解决将其从水体中移除的问题变得至关重要。这就要求用专门设计的工艺来补充现有的水处理技术，以消除微塑料。

To this end, various remediation technologies for water and wastewater treatment have been investigated, including membrane filtration, adsorption, coagulation-flocculation-sedimentation, bioremediation, and advanced oxidation processes. Next, in their review paper, Lu et al. analyze the advantages and disadvantages of each technique to ensure the efficient removal of microplastics from water sources (Lu et al., 2023).
为此，人们研究了用于水和废水处理的各种修复技术，包括膜过滤、吸附、混凝-絮凝-沉淀、生物修复和高级氧化工艺。接下来，在他们的评论论文中，Lu 等人分析了每种技术的优缺点，以确保有效去除水源中的微塑料（Lu 等人，2023 年）。

Moreover, a hybrid treatment approach using technologies like a membrane bioreactor or a combination of coagulation and membrane filtration is proving to be highly effective in microplastic removal (Adegoke et al., 2023).
此外，使用膜生物反应器等技术或混凝和膜过滤相结合的混合处理方法被证明在去除微塑料方面非常有效（Adegoke 等人，2023 年）。

4. Conclusion 4. 结论

This study presents the first evidence of microplastics in mine waters from two terminated underground mines in the largest coal region in the Czech Republic. It also compares the amount of microplastics in groundwater from nearby shallow wells. Microplastics were detected in all samples. Among 302 particles, 80 were confirmed to be polymers or additives used in plastic production. The identified microplastics included plastic coated paper, PET, PES, PP and TFE-PPVE, with fibers being the dominant shape and blue as the most common color. The presence of fibers suggests the existence of secondary microplastics originating from larger plastic pieces.
这项研究提供了捷克共和国最大的煤炭地区两个已终止的地下矿井的矿井水中微塑料的第一个证据。它还比较了附近浅井地下水中的微塑料含量。所有样本均检出微塑料。在302个颗粒中，有80个被确认是塑料生产中使用的聚合物或添加剂。鉴定出的微塑料包括塑料涂布纸、PET、PES、PP和TFE-PPVE，其中纤维是主要形状，蓝色是最常见的颜色。纤维的存在表明存在源自较大塑料片的次生微塑料。

The results show that microplastics contaminate waters to depths exceeding 700 m below the surface, supporting the hypothesis that almost all water sources, including mine waters, are affected by microplastics. Considering the expected limitations of microplastics in water for human consumption related to ISO 24187, the development of suitable technology for the removal of microplastics becomes imperative. Our findings therefore point to the importance of implementing effective microplastic remediation technologies when using mine waters as a source of drinking water.
结果表明，微塑料将水污染到地表以下超过700米的深度，这支持了几乎所有水源（包括矿井水）都受到微塑料影响的假设。考虑到与 ISO 24187 相关的人类消费水中微塑料的预期限制，开发合适的去除微塑料的技术变得势在必行。因此，我们的研究结果指出，在使用矿井水作为饮用水源时，实施有效的微塑料修复技术非常重要。

Author contributions 作者贡献

K.B., Conceptualization, investigation, methodology, writing - original draft, writing - review and editing, visualization, project administration; J.H., Conceptualization, methodology, investigation, formal analysis, writing - original draft, writing - review and editing, visualization; O.M., data curation, writing - review and editing; P.H., investigation, formal analysis; S.D., investigation; K.Č., investigation, validation, writing - review and editing; S.H., writing - review and editing, supervision. All authors have read and agreed to the published version of the manuscript.
K.B.，概念化，调查，方法论，写作 - 原始草稿，写作 - 审查和编辑，可视化，项目管理;J.H.，概念化，方法论，调查，形式分析，写作 - 原始草稿，写作 - 审查和编辑，可视化;O.M.，数据管理，写作 - 审查和编辑;P.H.，调查，形式分析;S.D.，调查;K.Č.，调查、验证、写作 - 审查和编辑;S.H.，写作 - 审查和编辑，监督。所有作者均已阅读并同意手稿的出版版本。

Institutional review board statement
机构审查委员会声明

Not applicable. 不適用。

Informed consent statement
知情同意声明

Not applicable. 不適用。

Declaration of Competing Interest
利益冲突声明

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
作者声明，他们没有已知的相互竞争的经济利益或个人关系，这些利益或关系可能会影响本文所报告的工作。

Funding 资金

This research was funded by the Student Grant Competition financed by VSB—Technical University of Ostrava within the project “Study of the occurrence of microplastics as an indicator of anthropogenic pollution in groundwater and mining waters” (no. SP2022/51) and project “Research and detection of micropollutants in drinking and waste water from sewage treatment plants.” (no. SP2023/045) and co-funded by the Technology Agency of the Czech Republic within the Environment for Life program-the project SS02030008 “centre of environmental research: Waste management, circular economy and environmental security”.
这项研究由VSB-俄斯特拉发技术大学资助的学生资助竞赛资助，在“研究微塑料的发生作为地下水和采矿水中人为污染的指标”项目（no.SP2022/51）以及“污水处理厂饮用水和废水中微污染物的研究与检测”项目。（编号。SP2023/045），并由捷克共和国技术局在“生命环境”计划中共同资助——该项目SS02030008“环境研究中心：废物管理、循环经济和环境安全”。

Acknowledgments 确认

Authors wish to thank DIAMO, s.p. for providing mine water samples and expert consultation and Alena Kašpárková and Kamila Průšová for her feedback.
作者感谢 DIAMO， s.p. 提供矿井水样本和专家咨询，并感谢 Alena Kašpárková 和 Kamila Průšová 的反馈。

Appendix. Supplementary materials
附录。补充材料

Download: Download Word document (2MB)

Data availability 数据可用性

Data will be made available on request.
数据将根据要求提供。

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Cited by (3) 被引文献（3）

Current research status on the distribution and transport of micro(nano)plastics in hyporheic zones and groundwater
微纳米塑料在低流变区和地下水中的分布与运移研究现状
2025, Journal of Environmental Sciences (China)
2025，环境科学学报
Micro(nano)plastics, as an emerging environmental pollutant, are gradually discovered in hyporheic zones and groundwater worldwide. Recent studies have focused on the origin and spatial/temporal distribution of micro(nano)plastics in regional groundwater, together with the influence of their properties and effects of environmental factors on their transport. However, the transport of micro(nano)plastics in the whole hyporheic zone-groundwater system and the behavior of co-existing substances still lack a complete theoretical interpretation. To provide systematic theoretical support for that, this review summarizes the current pollution status of micro(nano)plastics in the hyporheic zone-groundwater system, provides a comprehensive introduction of their sources and fate, and classifies the transport mechanisms into mechanical transport, physicochemical transport and biological processes assisted transport from the perspectives of mechanical stress, physicochemical reactions, and bioturbation, respectively. Ultimately, this review proposes to advance the understanding of the multi-dimensional hydrosphere transport of micro(nano)plastics centered on groundwater, the microorganisms-mediated synergistic transformation and co-transport involving the intertidal circulation. Overall, this review systematically dissects the presence and transport cycles of micro(nano)plastics within the hyporheic zone-groundwater system and proposes prospects for future studies based on the limitations of current studies.
The effect of water ozonation in the presence of microplastics on water quality and microplastics degradation
微塑料存在下水臭氧化对水质和微塑料降解的影响
2024, Science of the Total Environment
2024，整体环境科学
The occurrence of microplastics in water treatment plants poses a concern for the quality of treated water. When microplastics pass through water treatment plants, they can be oxidized, changing their surface characteristics and the quality of the treated water. This work aimed to investigate the impact of ozone and the association of ozone and hydrogen peroxide on five different microplastic particles that are commonly detected in water samples. The changes in the concentration of total organic carbon and the change in the pH of the water, the leaching of phthalic acid esters, as well as the changes in size and chemical changes in the structure of the tested microplastics were evaluated. The influence of ozonation time, water pH, and type of microplastics, as well as the influence of the addition of hydrogen peroxide, was analyzed. The effect of ozonation was an increase in DOC values ranging from 0.8 to 28 mg/L. The eluting substances included phthalic acid esters, plasticizers with a proven negative impact on organisms. The percentage loss of the surface area of the microplastic was in the range of 1.3 to 26.7 %. PE was more susceptible to degradation. LDIR analyzes were carried out to investigate the effect of O₃ and O₃/H₂O₂ treatments on the surface of MPs. This study demonstrated that MPs could change their physical and chemical characteristics if they are subjected to oxidation processes used in water treatment plants. The parameters of purified water change to unfavorable ones due to the leaching of additives. Although much research has been conducted on the occurrence of microplastics in treated water, awareness needs to be raised about the interactions between plastic particles and water treatment technology processes.
Microplastics in water: Occurrence, fate and removal
水中的微塑料：发生、归宿和去除
2024, Journal of Contaminant Hydrology
2024，污染物水文学报
A global study on tap water samples has found that up to 83% of these contained microplastic fibres. These findings raise concerns about their potential health risks. Ingested microplastic particles have already been associated with harmful effects in animals, which raise concerns about similar outcomes in humans. Microplastics are ubiquitous in the environment, commonly found disposed in landfills and waste sites. Within indoor environments, the common sources are synthetic textiles, plastic bottles, and packaging. From the various point sources, they are globally distributed through air and water and can enter humans through various pathways. The finding of microplastics in fresh snow in the Antarctic highlights just how widely they are dispersed. The behaviour and health risks from microplastic particles are strongly influenced by their physicochemical properties, which is why their surfaces are important. Surface interactions are also important in pollutant transport via adsorption onto the microplastic particles. Our review covers the latest findings in microplastics research including the latest statistics in their abundance, their occurrence and fate in the environment, the methods of reducing microplastics exposure and their removal. We conclude by proposing future research directions into more effective remediation methods including new technologies and sustainable green remediation methods that need to be explored to achieve success in microplastics removal from waters at large scale.

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