Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris

Summary

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

Abstract

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only recently have microplastics in freshwater systems, such as rivers and lakes, been quantified. Microplastic sampling at the surface usually consists of deploying drift nets behind either a stationary or moving boat, which limits the sampling to environments with low levels of suspended sediments and floating or submerged debris. Previous studies that employed drift nets to collect microplastic debris typically used nets with ≥300 µm mesh size, allowing plastic debris (particles and fibers) below this size to pass through the net and elude quantification. The protocol detailed here enables: 1) sample collection in environments with high suspended loads and floating or submerged debris and 2) the capture and quantification of microplastic particles and fibers <300 µm. Water samples were collected using a peristaltic pump in low-density polyethylene (PE) containers to be stored before filtering and analysis in the lab. Filtration was done with a custom-made microplastic filtration device containing detachable union joints that housed nylon mesh sieves and mixed cellulose ester membrane filters. Mesh sieves and membrane filters were examined with a stereomicroscope to quantify and separate microplastic particulates and fibers. These materials were then examined using a micro-attenuated total reflectance Fourier transform infrared spectrometer (micro ATR-FTIR) to determine microplastic polymer type. Recovery was measured by spiking samples using blue PE particulates and green nylon fibers; percent recovery was determined to be 100% for particulates and 92% for fibers. This protocol will guide similar studies on microplastics in high velocity rivers with high concentrations of sediment. With simple modifications to the peristaltic pump and filtration device, users can collect and analyze various sample volumes and particulate sizes.

Introduction

Plastic was first observed in the ocean as early as the 1930s¹. Recent estimates of marine plastic debris range from over 243,000 metric tons (MT) of plastic on the ocean's surface to 4.8-12.7 million MT of plastic entering the ocean from terrestrial sources annually^2,3. Early studies on marine plastic debris focused on macroplastics (>5 mm diameter) as they are easily visible and quantifiable. However, it was recently discovered that macroplastics represent <10% of plastic debris, by count, in the ocean, indicating that the overwhelming majority of plastic debris is micropl....

Protocol

1. Water Sample Collection

1. Collect water samples and water quality data of interest by boat where the river is well-mixed, ideally at locations where river stage or discharge is known (e.g., United States Geological Survey (USGS) gauging stations).²⁰ To assure that the water is well-mixed, guide the boat using a handheld meter immersed in the river to where conductivity stays relatively constant.
2. At the sampling sites, record location coordinates and depth. To find the 0.6-depth, simply multiply the total depth by 0.6. Measure water quality parameters of interest (e.g., turbidity, temperature, conductivit....

Results

To validate the recovery rates of this protocol, three samples (V₁-V₃) from Oso Bay, Corpus Christi, Texas (adjacent to the Texas A&M University Corpus Christi Campus), were spiked with 10 blue PE particulates (ranging from 50-100 µm in diameter) and 50 green nylon fibers of various lengths (Figure 3). Sample TSS was calculated (Section 2) and then the samples were filtered using the methods outlined in Sections 3-5. The blue PE part.......

Discussion

Microplastic collection using drift nets is the conventional method in environments like the ocean where both sediment and plastic concentrations are low, thus requiring large sample volumes. However, drift nets are not always practical or safe in rivers with high sediment loads and large floating or submerged debris. Additionally, it is not feasible to use a drift net when attempting to thoroughly capture and quantify microplastic materials, particularly fibers, as most nets used for plastic surveys have mesh sizes X.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
1L Cubitainer Containers, Low-Density Polyethylene	VWR	89094-140	Containers used to collect and store samples.
2-1/2" Clear Schedule 40 Rigid PVC Pipe	United States Plastic Corporation	34138	The PVC pipe used to make the device comes as an 2.43 m pipe. The pipe was then cut to the desired lengths for each section seperated by union joints. Section lengths were decided by predicting smaller pore sizes would clogg the device quicker. Longer sections were placed above the smaller pore sizes to collect and hold water to prevent needing to disassemble the device to change a filter while a sample remained in the device. For one filtration device one 18 in, one 12 in, and two 6 in peices are needed.
2-1/2" PVC SCH 40 Socket Union	Supply House	457-025	Union joints were glued to PVC pipe to house nylon sieves and mixed cellulose membranes.
Nylon 6 Woven Mesh Sheet, Opaque Off-White, 12" Width, 12" Length, 500 microns Mesh Size, 38% Open Area (Pack of 5)	Small Parts via Amazon	CMN-0500-C/5PK-05	Mesh sheets were cut into circles to match the diameter of the outer diameter of the PVC pipe. The edges were glued to esure no fraying would occur. The glue 's diamter should not extend into the inner diameter of the PVC so that it will not be affected during filtration.
Nylon 6 Woven Mesh Sheet, Opaque White, 12" Width, 12" Length, 100 microns Mesh Size, 44% Open Area (Pack of 5)	Small Parts via Amazon	B0043D1TB4	Mesh sheets were cut into circles to match the diameter of the outer diameter of the PVC pipe. The edges were glued to esure no fraying would occur. The glue 's diamter should not extend into the inner diameter of the PVC so that it will not be affected during filtration.
Nylon 6 Woven Mesh Sheet, Opaque White, 12" Width, 12" Length, 50 microns Mesh Size, 37% Open Area (Pack of 5)	Small Parts via Amazon	B0043D1SGA	Mesh sheets were cut into circles to match the diameter of the outer diameter of the PVC pipe. The edges were glued to esure no fraying would occur. The glue 's diamter should not extend into the inner diameter of the PVC so that it will not be affected during filtration.
Mixed Cellulose Ester Membrane, 0.45um, 142mm, 25/pk	VWR	10034-914	Mixed cellulose membrane filter with 0.45 um was used as the last filter. A large diameter was used to allow the filter to be folded into a cone to increase surface area of the filter to prevent clogging.
Metal Mesh Basket Tea Leaves Strainer Teapot Filter 76mm Dia 3pcs	Uxcell via Amazon	a15071600ux0260	The mesh basket used to provide extra support for the membrane filter to prevent tearing when pressure was applied by a vacuum pump.
1/2" PVC Barbed Insert Male Adapter	Supply House	1436-005	A vacuum adapter was added to allow vacuum filtration in the case of slow filtration due to high sediment concentration.
1/2 in. O.D. x 3/8 in. I.D. x 10 ft. PVC Clear Vinyl Tube	Home Depot	702229	Tubing used to connect the vacuum pump to the filtration device.
YSI Professional Plus Multiparameter Instrument with Quatro Cable	YSI	6050000	Handheld meter used to measure additional water quality parameters parameters (e.g., turbidity, temperature, conductivity, pH, and dissolved oxygen (DO)).
2100P Portable Turbidimeter	Hach	4650000	Handheld meter used to measure turbidity.
FEP-lined PE tubing	Geotech	87050529	Tubing used with perestaltic pump to collect water samples from desired depths.
Geopump Peristaltic Pump Series II	Geotech	91350123	Pump used to collected water samples.
MeiJi Techno EMZ-8TR Microscope	Microscope.com	EMZ8TR-PLS2	Microscope used analyze mesh sieves and membrane filters to quanitfy suspect microsplastics.
Nicolet iS10 FTIR Spectrometer	Thermo Electron North America	912A0607	FTIR used to analyze suspect microplastics.
Nicolet iN5 FTIR microscope	Thermo Electron North America	912A0895	FTIR microscope used to analyze suspect microplastics.
Germanium (Ge) ATR	Thermo Electron North America	869-174400	Geranium ATR accessory used along with the Nicolet iN5 FTIR microscope to analyze suspect microplastic.
Aluminum EZ-Spot Micro Mounts (Pkg of 5)	Thermo Electron North America	0042-545	Microscope slides used along with the Nicolet iN5 FTIR microscope to analyze suspect microplastic.
Aluminum Coated Glass Sample Slides	Thermo Electron North America	0042-544	Microscope slides used along with the Nicolet iN5 FTIR microscope to analyze suspect microplastic.