![Page 1: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/1.jpg)
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
510
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95 1…
1…
110
0
40
80
120
160
200
90 110 130 150 170 190
![Page 2: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/2.jpg)
![Page 3: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/3.jpg)
![Page 4: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/4.jpg)
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
![Page 5: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/5.jpg)
![Page 6: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/6.jpg)
![Page 7: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/7.jpg)
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
![Page 8: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/8.jpg)
Inductively Coupled Plasma – Mass Spectrometry (ICP-MS)
Liquid sample
![Page 9: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/9.jpg)
Single Particle (SP) - ICP-MS
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
Au+
![Page 10: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/10.jpg)
![Page 11: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/11.jpg)
Single Particle ICP-MS of a Au NP suspension
0
50
100
150
200
250
300
350
400
0 5000 10000 15000 20000 25000 30000
Sign
al p
er d
wel
l tim
e (5
mse
c)
Time / msec
Degueldre, C.; Favarger, P. Y.; Wold, S. Anal. Chim. Acta 2006, 555, 263-268.
![Page 12: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/12.jpg)
0
5
10
15
20
25
30
35
40
45
50
21 25 29 33 37 41 45 49 53 57 61 65 69 73 77 81 85 89 93 97
NU
MB
ER O
F N
P C
OU
NTE
D
PARTICLE SIZE / NM
Particle Size Distribution
𝑑𝑖,𝐴𝑔𝑁𝑃 =3 6𝑚𝑖,𝐴𝑔𝑁𝑃
𝜋 𝜌
![Page 13: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/13.jpg)
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
![Page 14: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/14.jpg)
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)
![Page 15: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/15.jpg)
Background
unrealistic and simplified conditions
high concentrations of AgNPs
single-species cultures or pre-treated estuarine & freshwater
communities
short-term experiments
![Page 16: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/16.jpg)
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
![Page 17: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/17.jpg)
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)
![Page 18: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/18.jpg)
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
![Page 19: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/19.jpg)
0
20
40
60
80
100
120
140
160
0 100 200 300 400 500
Cou
nts
pe
r 1
0 m
s f
or
m/z
10
7
Time (s)
0
20
40
60
80
100
120
140
160
0 100 200 300 400 500
Cou
nts
pe
r 1
0 m
s f
or
m/z
10
7
Time (s)
a b
c d
0
5
10
15
20
25
30
35
5 15 25 35 45 55 65 75 85 95 105
Fre
qu
en
cy
Size (nm)
0
5
10
15
20
25
30
35
5 15 25 35 45 55 65 75 85 95 105
Fre
qu
en
cy
Size (nm)
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
![Page 20: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/20.jpg)
𝑚𝐴𝑔𝑁𝑃𝑠𝑡𝑑 = 𝑘. ത𝑞𝐴𝑔𝑁𝑃
𝑘 =𝑚𝐴𝑔𝑁𝑃
𝑠𝑡𝑑
ത𝑞𝐴𝑔𝑁𝑃
𝑚𝑖,𝐴𝑔𝑁𝑃 = 𝑘. 𝑞𝑖,𝐴𝑔𝑁𝑃
0
40
80
120
160
200
0 100 200 300 400
Co
un
ts p
er 1
0 m
s fo
r m
/z 1
07
Time (s)
Seawater sample
*
******
**
*
** *
𝑚n,𝐴𝑔𝑁𝑃 = 𝑘. 𝑞n,𝐴𝑔𝑁𝑃
...
...
...
Std AgNP 60 nm
ത𝑞𝐴𝑔𝑁𝑃
![Page 21: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/21.jpg)
𝑑𝑖,𝐴𝑔𝑁𝑃 =3 6𝑚𝑖,𝐴𝑔𝑁𝑃
𝜋 𝜌
ҧ𝑑𝐴𝑔𝑁𝑃
𝜀𝑛 =𝑛𝑑𝑒𝑡𝑛𝑖𝑛𝑗
NP number concentration determination
![Page 22: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/22.jpg)
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
![Page 23: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/23.jpg)
25
30
35
40
45
50
55
0 1000 2000 3000
Exposure time (min)
AgBPEI 60 nm AgBPEI 40 nm
AgPVP 60 nm AgPVP 40 nm
ҧ𝑑𝐴𝑔𝑁𝑃
(nm
) 9 analytical runs
![Page 24: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/24.jpg)
160
510
1660
2990
0
1E-12
2E-12
3E-12
2530354045505560657075808590
Exp
osu
re T
ime
(min
)
Ag
mas
s in
AgN
Ps
(g)
Size (nm)
AgPVP 60 nm
160
510
1660
29900
2E-12
4E-12
6E-12
2530354045505560
exp
osu
re t
ime
(min
)
Ag
in A
gNP
s (g
)
Size (nm)
AgPVP 40 nm
160
510
1660
29900
1E-12
2E-12
3E-12
4E-12
2530354045505560
Exp
osu
re T
ime
(min
)
Ag
in A
gNP
s (g
)
Size (nm)
AgBPEI 40 nm
160
510
1660
2990
0
1E-12
2E-12
3E-12
2530354045505560657075808590
Exp
osu
re T
ime
(min
)
Ag
mas
s in
AgN
Ps
(g)
Size (nm)
AgBPEI 60 nm a
c d
b
![Page 25: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/25.jpg)
Methodology
Whole-community approach challenges
Mesocosm experiment “Cretacosmos- HCMR”
3-m3 incubations, 3 replicates
33 days
biotic and abiotic factors studied
![Page 26: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/26.jpg)
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)
![Page 27: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/27.jpg)
13
57
911
26
13
57
911
26
35
40
45
50
55
60
Ave
rage
AgN
P s
ize
(nm
)
0
200
400
600
800
1000
0 5 10 15 20 25 30 35
Ag
Co
nce
ntr
atio
n (
ng/
L)
Days of Incubation
b
c
- 0.24
- 0.35
- 0.50
- 0.68
- 0.91
- 1.19 Ag m
ass equ
ivalence (fg)
a
d
Ag diameter size (nm)
Ag
con
cen
trat
ion
(n
g/L
)A
gNP
nu
mb
er c
on
cen
trat
ion
(# p
er m
L)
Days of incubation
![Page 28: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/28.jpg)
Temporal changes in Plankton abundances
Cyanobacterial growth
was inhibited
![Page 29: The analysis of Metal-containing Engineered Nanoparticles](https://reader031.vdocuments.us/reader031/viewer/2022012501/617ac8c3294fb655bc7119b0/html5/thumbnails/29.jpg)
• 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