Spiros A. Pergantis

Environmental Chemical Processes Laboratory,

Dept. of Chemistry, University of Crete

Voutes Campus, Heraklion, 70013, Greece

The analysis of Metal-containing Engineered Nanoparticles

using SINGLE PARTICLE ICP-MS

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Presentation Outline

• Brief intro to sp ICP-MS

• Application of sp ICP-MS for Ag NP detection in a marine mesocosm experiment

• Limitations of sp ICP-MS

• NP Sizing using HDC, FFF, and ion mobility with sp ICP-MS

• NP characterization using modified sp ICP-MS approaches

• Development and application of single cell ICP-MS analysis

Inductively Coupled Plasma – Mass Spectrometry (ICP-MS)

• Analysis of sample with a heavy or complex matrix (i.e. seawater, biological samples)

• Routine analysis for screening purposes (high throughput with minimum sample preparation)

• Not labor intensive

• Sensitive with LOD adequate for realistic environmental concentrations

Inductively Coupled Plasma – Mass Spectrometry (ICP-MS)

Liquid sample

Single Particle (SP) - ICP-MS

Au+

Au+

Au+

Au+

Au+

Au+

Au+

Au+

Au+

Au+

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Au+

Au+

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Au+

Au+

Au+

Single Particle ICP-MS of a Au NP suspension

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al p

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Degueldre, C.; Favarger, P. Y.; Wold, S. Anal. Chim. Acta 2006, 555, 263-268.

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Particle Size Distribution

𝑑𝑖,𝐴𝑔𝑁𝑃 =3 6𝑚𝑖,𝐴𝑔𝑁𝑃

𝜋 𝜌

Anastasia Tsiola 1,2, Paraskevi Pitta 2, Margarita Kagiorgi 2, Ioanna Kalantzi 2,

Kyriaki Mylona 2,3, Stella Psarra 2, Ioulia Santi 1,2, Claudio Toncelli 2,3,

Spiros A. Pergantis 3, Manolis Tsapakis 2

1 University of Crete, Department of Biology2 Hellenic Centre for Marine Research, Institute of Oceanography

3 University of Crete, Department of Chemistry

Effects of silver nanoparticle exposure at the ng L-1 level

on marine microbial plankton dynamics

Background

engineered NPs are used in a wide range of applications

AgNPs release in coastal zone is expected to increase in the near future

consequences on aquatic biota have been already determined

apoptosis

ROS production

decreased viability and growth rates

decreased chlorophyll content

alterations in community composition

alterations in protein expression patterns

Silver nanoparticles of 60 nm diameter (TEM; Nanoxact,

No MRL1009)

Background

unrealistic and simplified conditions

high concentrations of AgNPs

single-species cultures or pre-treated estuarine & freshwater

communities

short-term experiments

Aim – AQUA-NANO project

potential effects of AgNPs in a natural planktonic communityno pre-filtration step, total marine plankton incubated

environmental relevant concentrations of AgNPs100-1000 ng Ag L-1

long-term effectsover a month

interaction effects on the genus or species level, when possible

considering the ambient physico-chemical parameters

Methodology

Analytical challenges – FI sp ICP MS

very dilute samples

very complex matrix

large number of samples

Whole-community approach challenges – mesocosms

microbial planktonic members present

constant interplay with highly-variable ambient conditions (pH,

temperature, organic matter, O2 and others)

single particle Inductively Coupled Plasma Mass Spectrometry

Ar gas

3-5 mmdilution zone

HPLC pump

Injector (20 μL loop)

di water

10-50 μL min-1

0.8 mL min-1

Fused silica capillary

Toncelli et al., 2016

Methodology

Analytical challenges

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Methodology

Analytical challenges

AgBPEI 60 nm NPs at a concentration of 200 ng Ag L-1. Histogram is the sum of 3 injections.

dH2O seawater

𝑚𝐴𝑔𝑁𝑃𝑠𝑡𝑑 = 𝑘. ത𝑞𝐴𝑔𝑁𝑃

𝑘 =𝑚𝐴𝑔𝑁𝑃

𝑠𝑡𝑑

ത𝑞𝐴𝑔𝑁𝑃

𝑚𝑖,𝐴𝑔𝑁𝑃 = 𝑘. 𝑞𝑖,𝐴𝑔𝑁𝑃

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𝑚n,𝐴𝑔𝑁𝑃 = 𝑘. 𝑞n,𝐴𝑔𝑁𝑃

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Std AgNP 60 nm

ത𝑞𝐴𝑔𝑁𝑃

𝑑𝑖,𝐴𝑔𝑁𝑃 =3 6𝑚𝑖,𝐴𝑔𝑁𝑃

𝜋 𝜌

ҧ𝑑𝐴𝑔𝑁𝑃

𝜀𝑛 =𝑛𝑑𝑒𝑡𝑛𝑖𝑛𝑗

NP number concentration determination

Part of the “Aqua-Nano” project ( http://aqua-nano.hcmr.gr/ )

poly(vinylpyrrolidone), PVP (Branched poly(ethylenimine)), BPEI

AgPVP AgBPEI

40 nm

60 nm

Control

200 ppt Ag for each Ag NP type

15 tanks in total

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Exposure time (min)

AgBPEI 60 nm AgBPEI 40 nm

AgPVP 60 nm AgPVP 40 nm

ҧ𝑑𝐴𝑔𝑁𝑃

(nm

) 9 analytical runs

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AgPVP 40 nm

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Methodology

Whole-community approach challenges

Mesocosm experiment “Cretacosmos- HCMR”

3-m3 incubations, 3 replicates

33 days

biotic and abiotic factors studied

Mesocosm experiment

Microcosm experiments (Toncelli et al., 2017 and Tsiola et al., 2017)

coastal seawater from the Cretan Sea, April-May 2015

branched poly(ethyleneimine) AgNPs, 60 nm

100 ng Ag L-1 spiking concentration per day for 10 days (step-wise)

final maximum concentration: 1000 ng Ag L-1

3x BPEI of 60 nm diameter

3x controls (C)

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Temporal changes in Plankton abundances

Cyanobacterial growth

was inhibited

• The amount of a lysogeny-related gene increased and viral auxiliary metabolic

genes that are involved in cyanobacterial photosynthesis decreased, revealing a

damaged photosynthetic potential after AgNP exposure.

• Microbial plankton was significantly affected due to both increased dissolved silver

ions and decreased AgNP size.

• The presence of AgNPs alters the functioning of the marine food web by hampering

important viral and bacterial processes.

General Conclusions

Environ. Sci.: Nano, 2018,5, 1965-1980