JPI/Healthy Oceans and ICES open session on microplastics€¦ · ICES ANNUAL SCIENCE CONFERENCE 2016 • BASEMAN - Defining the baselines and standards for microplastics analyses

ICES ANNUAL SCIENCE CONFERENCE 2016

JPI/Healthy Oceans and ICES open session on microplastics


Why is this topic important?

JPI-O pilot action “Ecological Aspects of Microplastics”



PlasticsEurope, 2013

1.7MioT

288MioT

• We use plastics



• We discard plastics



„Prä-Plastik Ära“

mellumrat.de

• We act irresponsible



• We have limited knowledge... on pathways, sources and sinks



• We have limited knowledge... on persistence

• Please note: Plastics are not part of biogeochemical cycles



MacroplasticsMicroplastics



• We have limited knowledge... MP numbers & identities ...because...

• we use different methods for sampling, extraction, purification AND identification

Wright et al., Environmental Pollution (2013)



• We have limited knowledge... on effects



OK...

but do we really know quantities and what is causing “harm” and how?

We need facts (and standardizations)!

• “Properties and quantities of marine litter…cause harm to the coastal and marine environment” (known as ‘Descriptor 10’).

• This definition includes microparticles (particularlymicroplastics)“

MSFD


• BASEMAN - Defining the baselines and standards for microplastics analyses in European waters (Gunnar Gerdts, AWI, Germany)

• PLASTOX - Direct and indirect ecotoxicological impacts of microplastics on marine organisms (Andy Booth, SINTEF, Norway)

• WEATHERMIC - How microplastic weathering changes its transport, fate and toxicity in the marine environment (Annika Jahnke, UFZ, Germany)

• (EPHEMARE - Ecotoxicological effects of microplastics in marine ecosystems)

• MikrOMIK - Microplastic as vector for microbial populations in the ecosystem of the Baltic Sea (Sonja Oberbeckmann, IOW, Germany)

JPI-O pilot action “Ecological Aspects of Microplastics”Start January 2016


Defining the BASElines and standards for Microplastics ANalyses in European Waters (BASEMAN)

Gunnar GerdtsMicrobial EcologyShelf Seas Systems EcologyAlfred Wegener Institute Helmholtz Centre for polar and marine research27483 Helgoland

• 24 partners from 11 countries (AWI in lead)• 5 workpackages

WP1 Defining baselines for all relevant identification approachesParticipating institutions

UBAY, UGOT, UDC, AWI, ICBM, NILU, GEOMAR, Rap-ID, IMMM, CNR-IAMC, IVL, CNRS-LOV

Objectives

• To develop and provide a MP reference kit for baseline definition and spiking of samples

• To characterize the effects of artificial/natural weathering on reference MPs

• To generate a spectra database (FT-IR, Raman, Pyrolysis-GCMS, HySpex) for pristine and weathered reference particles

• To evaluate the strengths and limitations of different analytical techniques without interferences of environmental matrices

• To develop methods with submicron capability in order to define a reasonable (methodological) lower size limit of MP

WP 2 Preparation of standardized test samples for inter-lab comparisonsParticipating institutions

AWI, ICBM, NIFES

Objectives

• To generate and provide standardized MP-spiked test samples (sediment, plankton, biota) for inter-lab comparisons (feeding into WP3)

• To generate and provide standardised test samples (sediment, plankton, biota) for inter-method comparisons (feeding into WP3)

WP 3 Inter-lab and inter-method comparisons

Participating institutions

AWI, ICBM, UBAY, NIFES, NIVA, GEOMAR, CNRS-LOV, TUT, IEO, UGOT, VUT, SYKE, Rap-ID, IMMM, CNR-IAMC, NOVA.ID FCT, IVL, IPMA, UDC, UL

Objectives

• To analyse MP-spiked reference samples (sediment, plankton, biota) according to the current analytical workflows in the participating labs with respect to identification, quantification and sizing of MP

• To analyse environmental samples (sediment, plankton, biota) provided by WP4 according to the current analytical workflows in the participating labs with respect to identification, quantification and sizing of MP

• To optimize the extraction of MP from sediment

• To optimize the purification of MP from sediment, plankton and biota in respect to matrix disintegration/removal and polymer preservation

WP 4 Sampling methodologies for MPs in the marine environment: standardization, suitability and intercomparison


IEO, GMIT, NIVA, UBAY, OGS, CNRS-LOV, AWI, ICBM, IMMM, CNR-IAMC, NOVA.ID FCT, TUT, IVL, IPMA, SAHFOS, UDC

Objectives

• To develop standardized methods for sampling MPs in the water column and sediments

• To identify marine biota species appropriate for MPs monitoring in European regions

• To evaluate alternative methods for sampling MPs in the marine environment, particularly those deployable from platforms of opportunity

• To intercalibrate sampling methods for MPs of varying size

• To facilitate interlab comparisons and analyses of environmental samples of MPs

• To develop validated conversion coefficients and harmonized reporting units for MPs

WP 5 Coordination, Integration and Synthesis


AWI & all

Objectives

• To provide “best practice” and “best compromise” SOPs

• To advice stakeholders with respect to MP sampling, detection & analysis

Some preliminary results

Defining the BASElines and standards for Microplastics ANalyses in European Waters (BASEMAN)

WP1 Defining baselines for all relevant identification approachesObjectives

• To develop and provide a MP reference kit for baseline definition and spiking of samples

Methodological comparisons

Methodological thresholds

Best compromise (→ MSFD) & best practise

Different polymers(different size classes)( 1 lab in charge)

• Cryo-milling• Thieving• Size analysis• Polymer-kit prep.

Different environmental samples (3 labs in charge) • Kit + Sediment (3 x (3+1)) • Kit+ Plankton (3 x (3+1))• Kit + Biota (3 x (3 +1))

Participating labs• Optical microscopy• µFT-IR• Raman• PyGCMS

Problems…

• High organic (inorganic) load

• Interference with analytical approaches

• Overload of analytical filters

• Agressive versus inagressive agents

• Effectiveness in terms of time and costs (→MSFD)

WP 3 Inter-lab and inter-method comparisonsTo optimize the purification of MP from sediment, plankton and biota in respect to matrix disintegration/removal and polymer preservation

Solutions…

• Gentle, plastics-preserving treatments (e.g. enzymes)

• Effectiveness in terms of costs (→MSFD)

• Technical grade agents

• “used” (in Mani et al. (2016) as pers. comm)

• “publication” soon

Lorenz, 2014; Ladehoff, 2015


Challenges

• Time consuming: Need for optimization

• Manual steps: Risk of contamination

Lorenz, 2014; Ladehoff, 2015


Challenges and plans…


• Manual steps: Risk of contamination

• Development of a “reactor”

• All steps (incl. incubation) in one containment

• Fill & drain through 10 µm filters (pump; vacuum/pressure)

• Processing of multiple samples

• Monitoring (→MSFD)

©Lorenz


Challenges and plans…


• Overload of analyses filters: Failure in final analyses

• Usage of “FlowCam”

• Process control

• Estimation of filtration volume for µFT-IR analysis

©Lorenz


Bruker Tensor 27

Bruker Tensor 27 with ATR

Bruker Hyperion 3000 (FPA 64 x 64)Norhof LN2 (liquid nitrogen filling system)

• Sortable (single) particles (> 500 µm): ATR-FT-IR

• Bulk samples (< 500 µm; extracts from plankton, sediment, animalhomogenates, sea ice….). Chemical imaging of whole filter areas: µFT-IR

µFT-IR analysis

From 09/2016 two Hyperion systems

FT-IR MikroscopeFPA Detector (64x64)

• ~ 106 Spektra• 1 Pixel = 10.74 µm

µFT-IR analysis

• Sortable (single) particles (> 500 µm): ATR-FT-IR

• Bulk samples (< 500 µm; extracts from plankton, sediment, animalhomogenates, sea ice….). Chemical imaging of whole filter areas: µFT-IR

VIS Image Chemical Image(polymer)

FT-IR spectrum

Polymer-signatures

µFT-IR analysis

• 100% Microplastics!• Re-analysis by µFT-IR

• 1.4 %: „Plastics“• Quartz (sand)

dominating the „MP particles“

Quartz-signature

Polymer-signature

„…up to 70% of particles thatvisually resemble microplasticsare not confirmed as plastics byFT-IR spectroscopy…“ Hidalgo-Ruz

et al. 2012

µFT-IR analysis

WP 3 Inter-lab and inter-method comparisonsTo analyse environmental samples (sediment, plankton, biota) ...according to the current analytical workflows in the participating labs with respect to identification, quantification and sizing of MP

Problems…

• ~ no commercial pipeline (hardware, software)

• Raman: Horiba, Rap-ID (to be evaluated in BASEMAN)

Solutions...

• Development of an automated pipeline for µFT-IR (spectrum by spectrum and pixel by pixel...; ~ 106 spectra)

• Bruker Opus Macros

• Image analysis by Python and SimpleITKscripts

Light Microscopy µFT-IR and Image analysis

Light Microscopy µFT-IR and Image analysis

„Signature-image“


Solutions...

• Development of an automated pipeline for µFT-IR

• Bruker Opus Macros

• Image analysis by Python and SimpleITKscripts

Identities!

Numbers!

Sizes!


Kantenlänge [µm]

0 50 100 150 200 250

N

0

100

200

300

400

Kantenlänge [µm]

0 50 100 150 200 250

N

0

100

200

300

400

y = 6012 * e-0.26 *x

adj r2 = 0.9846

< 10.74 µm ?????

10.74 µm = 1 Pixel

WP1 Defining baselines for all relevant identification approaches – to be continued....

Kantenlänge [µm]

0 50 100 150 200 250

N

0

100

200

300

400

Kantenlänge [µm]

0 50 100 150 200 250

N

0

100

200

300

400

y = 6012 * e-0.26 *x

adj r2 = 0.9846

< 10.74 µm ?????

To develop methods with submicron capability in order to define a reasonable (methodological) lower size limit of MP

Documents

JPI/Healthy Oceans and ICES open session on microplastics€¦ · ICES ANNUAL SCIENCE CONFERENCE 2016 • BASEMAN - Defining the baselines and standards for microplastics analyses