ASTM D8333-20

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation:D8333 −20
Standard Practice for
Preparation of Water Samples with High, Medium, or Low
Suspended Solids for Identification and Quantification of
Microplastic Particles and Fibers Using Raman
Spectroscopy, IR Spectroscopy, or Pyrolysis-GC/MS
This standard is issued under the fixed designation D8333; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 1.5 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
1.1 This practice provides for the sample preparation of
responsibility of the user of this standard to establish appro-
collected water samples with high, medium, or low suspended
priate safety, health, and environmental practices and deter-
solids to determine the presence, count, polymer type, and
mine the applicability of regulatory limitations prior to use.
physical characteristics of microplastic particles and fibers. It
1.6 This international standard was developed in accor-
has been designed for the preparation of samples collected
dance with internationally recognized principles on standard-
from drinking water, surface waters, wastewater influent and
ization established in the Decision on Principles for the
effluent (secondary and tertiary), and marine waters using
Development of International Standards, Guides and Recom-
collection practice (Practice D8332). This practice is not
mendations issued by the World Trade Organization Technical
limited to these particular water matrices; however, the appli-
Barriers to Trade (TBT) Committee.
cability of this practice to other aqueous matrices must be
demonstrated.
2. Referenced Documents
1.2 This practice consists of a wet peroxide oxidation 2
2.1 ASTM Standards:
followed by progressive enzymatic digestion to the extent
D883 Terminology Relating to Plastics
necessary to remove interfering organic constituents such as
D1193 Specification for Reagent Water
cellulose, lipids and chitin that are typically found in abun-
D8332 Practice for Collection of Water Samples with High,
dance in water matrices of samples with high to medium
Medium, or Low Suspended Solids for Identification and
suspended solids such as wastewater influent. For water
Quantification of Microplastic Particles and Fibers
samples with low suspended solids, such as but not limited to
drinking water and tertiary treated wastewater, the oxidation
3. Terminology
and digestion steps may not be necessary.
3.1 Definitions:
1.3 Water samples prepared using this practice are suitable
3.1.1 For definitions of terms used in this standard, refer to
for analysis utilizing either Pyrolysis-GC/MS methods for
Terminology D883.
qualitative identification and mass quantitation, or IR spectros-
3.2 Definitions of Terms Specific to This Standard:
copy or Raman spectroscopy for identifying the quantity
3.2.1 effluent, n—any stage of treated wastewater.
(number count) and composition (polymer type) of microplas-
3.2.2 influent, n—raw sewage entering a wastewater treat-
tic particles. If desired, microplastic particle size and shape
ment facility.
may be ascertained with appropriate instruments such as a
scanning electron microscope (SEM) and microscopy tech-
3.2.3 microplastic, n—any solid, synthetic organic poly-
niques.
meric material to which chemical additives or other substances
may have been added, which are particles <5 mm in their
1.4 Units—The values stated in SI units are to be regarded
largest dimension, and fibers no longer than 15 mm in length
as standard. No other units of measurement are included in this
with an aspect ratio of at least 30:1 and <500 µm in its smallest
standard.
dimension.
This practice is under the jurisdiction of ASTM Committee D19 on Water and
is the direct responsibility of Subcommittee D19.06 on Methods for Analysis for For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Organic Substances in Water. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Current edition approved July 15, 2020. Published August 2020. DOI: 10.1520/ Standards volume information, refer to the standard’s Document Summary page on
D8333-20. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D8333−20
3.2.4 surface waters, n—a water body with its surface in 6. Reagents and Materials
contact with the ambient atmosphere, examples which include
6.1 PurityofwatershallbereagentwaterasdefinedbyType
lakes, rivers, and streams.
IV of Specification D1193.
3.2.5 suspended solids, n—refers to all matter which remain
6.2 Conical centrifuge tubes, 50 mL, with lids.
in suspension in water media and that are removed by a 0.45
6.3 Temperature controlled orbital test tube shaker,or
µm filter.
similar test tube rocker.
4. Summary of Practice
6.4 Laboratory centrifuge and rotor, suitable for containing
50 mL conical centrifuge tubes.
4.1 Thispreparationpracticeisapplicableforwatersamples
with high to very low suspended solids. All water samples
6.5 Analytical balance (precision to 0.1 mg).
shouldbecollectedusingprocedureinPracticeD8332,Section
6.6 Digital pH meter.
7. For those water samples with high to medium suspended
solids, it consists of a wet peroxide oxidation followed by
6.7 Protease Reagent—Proteasefrom Bacilius licheniformis
progressive enzymatic digestion of sieve contents. It is appro-
– ≥ 2.4 units/g with storage temperature from 2–8°C.
priate for collected raw wastewater influent, secondary waste-
6.8 Lipase Reagent—Lipase from Aspergillus oryzae –
water effluent, and surface waters samples which are high in
solution, ≥ 100 000 units/g with storage temperature from
suspended solids to remove interfering organic constituents
2–8°C.
such as cellulose, lipids, and chitin that are typically found in
abundance in these water matrices. The preparation steps are 6.9 Hydrogen peroxide,30%.
completed in the following sequence:
6.10 Tris HCl buffer1M,pH8.
4.1.1 Wet peroxide digestion (1 hr),
6.11 Methanol.
4.1.2 Centrifuge and decant excess liquid (15 min),
4.1.3 CellulosedigestionwithmodifiedSchweizer’sreagent
6.12 Graduated cylinder, 25 mL, glass.
(5 min),
6.13 Copper (II) hydroxide, granular.
4.1.4 Centrifuge and decant excess liquid (5 min),
6.14 Ammonium hydroxide,30%
4.1.5 Schweizer’s reagent quench with 30 % NH OH (5
min),
7. Procedure
4.1.6 Buffer sample to pH 8 with Tris-HCl (5 min),
4.1.7 Simultaneous protease and lipase digestion (20 hrs),
7.1 High to Medium Suspended Solids:
4.1.8 Centrifuge and decant excess liquid (5 min),
7.1.1 Transfer collected sieve materials [from the 0.25 L
4.1.9 Water wash (5 min), and
glass collection jar (petri dish) using procedure in Practice
4.1.10 Centrifuge and decant excess liquid, add 10 mL of
D8332, 7.1.8 and 7.2.7] into a 50 mL centrifuge tube using 25
CH OH.
mL of 30% H O . Begin wet peroxide oxidation of organic
2 2
matter by capping and placing centrifuge tube in the test tube
4.2 Samples collected using Practice D8332 are suitable for
rocker for 60 minutes at 20 rpm.
preparation using this practice.
7.1.2 Centrifuge contents at 5000 rpm for 3 minutes to
create a sediment pellet within the tube.
5. Significance and Use
7.1.3 Using a disposable glass pipette, remove as much of
5.1 Large volumes of water are required to be sieved for
the remaining hydrogen peroxide solution as possible from the
accurate quantification of microplastics. Water with high to
sample without disturbing the sediment fraction.
medium content of suspended solids can lead to an excess of
7.1.4 Prepare a modified Schweizer’s reagent consisting of
inorganic and organic background material which can interfere
2.5 g copper (II) hydroxide added to 100 mL of 30%
with the ability to conduct reliable analyses. The presence of
ammonium hydroxide. Stir contents for 10 minutes to dissolve.
this background material can often impede the ability to
Contents will not dissolve completely due to an excess of
accurately discern, distinguish and identify the number of
copper hydroxide.
microplastic particles in solution.
7.1.5 Initiate cellulose digestion by adding 40 mL of the
5.2 The digestion described in this
...