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href="#fb3_img_img_3ed9f46b-8612-5d0e-a64d-19d55fc6c4c0.jpg" alt="Schematic illustration of possible exposure pathways of MPs into freshwater environments."/>

      1.4.1 Sampling of Microplastic

      1.4.1.1 Water Samples

Schematic illustration of techniques reported in the literature for identifying MPs in sediment and water samples.

      1.4.1.2 Sediment Samples

      The sampling of sediment samples can be separated into the collection from the shoreline and bottom of the river or lake. For shore sediments, sampling strategies are transected sampling perpendicular, random sampling, and sampling in single squares or parallel to the water. Most studies applied the grid sampling method with depths of 2–5 cm on the surface layer (Jiang et al. 2019; Klein et al. 2018). Frame and corers are usually used to determine the sampling area. Non‐plastic tools such as scoop, trowels, or shovels, and non‐plastic sampling vessels are required (Alam et al. 2019; Jiang et al. 2019; Peng et al. 2018). Bottom sediment from the riverbed or lakebed can be carried out with grab samplers such as Ekman or Van Veen grabs or corers (Alam et al. 2019; Fan et al. 2019; Ta et al. 2020c; Wang et al. 2017). The sediment samples collected by grab methods are usually disturbed, therefore this is suitable for surface layer (top 5 cm) or bulk sampling. Conversely, sampling by cores allows determining MP depth profiles and undisturbed surface and depth layers. Nevertheless, the number of samples that can be collected is limited. According to Dris et al. (2018), river bottom sediments are mostly collected by grabs, while corers or grabs are used for lake bottom sediments. The number of MPs is usually normalized to the sediment volume or weight, and sampling area.

      1.4.2 Sample Preparation

      1.4.2.1 Extraction of Microplastics

      Due to the complex nature of the sediment, MPs in the samples must be extracted from sample matrices. The density separation is widely applied to extract MPs from sediment samples. The sediment is dried prior to mixing with a concentrated salt solution. After a period of agitation, MPs and light particles float to the surface or stay suspended, whereas heavy particles settle down (Klein et al. 2018). Many studies extract MPs by using sodium chloride (NaCl) solution since this is inexpensive and environmentally friendly (Alam et al. 2019; Campanale et al. 2020; Free et al. 2014; Mani et al. 2015). However, the density of NaCl solution (~1.2 g/cm3) cannot extract some polymers such as Polyvinyl chloride (PVC), Polyethylene terephthalate (PET), polycarbonate, and polyurethane. Therefore, sodium iodide (NaI), sodium zinc chloride (ZnCl2), and sodium polytungstate (Na2WO4) are viable choices (Ballent et al. 2016; Ta and Babel 2019; Yin et al. 2020). Conversely, MPs in water samples are easily filtered and separated during the sampling step (Dris et al. 2018).

      1.4.2.2 Removal of Organic Debris

      1.4.3 Identification of Microplastic

      1.4.3.1 Visual Sorting

      In most studies, visual sorting is the first step to separate MPs from samples before identification of the polymer type. Large MPs (> 1 mm) can be recognized by the naked eye (Anderson et al. 2017), while smaller particles are identified using dissection microscopes (Faure et al. 2015; Mani et al. 2015) or scanning electron microscopy (SEM) (Eriksen et al. 2013; Su et al. 2016). This step requires experienced researchers and good optical quality of the microscope. However, identification of all particles is difficult if they are smaller than a certain size, if they are unable to be distinguished visually or cannot be managed with forceps due to their minuteness. Thus, visual sorting is time‐consuming and easy misidentification or underestimation of MPs is possible. Recently, another visual identification method using fluorescence was applied to detect and quantify small MPs. In most studies, Nile Red (NR) was used and dissolved in different solvent solution such as acetone, chloroform, and

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