Article

pubs.acs.org/ac

Size Discrimination and Detection Capabilities of Single-Particle

ICPMS for Environmental Analysis of Silver Nanoparticles
Jani Tuoriniemi, Geert Cornelis, and Martin Hassellöv*
Department of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, Sweden
*
S Supporting Information

ABSTRACT: The detection capabilities of single particle

inductively coupled plasma-mass spectrometry (spICPMS)
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with respect to particle size and number concentrations are

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investigated for the case of silver nanoparticles (ca. 20−80

nm). An iterative algorithm was developed where particle
measurement events were distinguished as outliers from the
more continuous dissolved ion signal if the measured intensity
was more than five times the standard deviation of the whole
data set. The optimal dwell time for 40−80 nm particles,
limiting both incomplete and multiple particle events, was 5
ms. The smallest detectable particle size (ca. 20 nm) is mainly
limited by the overlap of particle events and dissolved signal that increases with noise on both signals. The lowest measurable
number concentration is limited by the relative frequency of erroneously identified particle events, a limit that can be reduced by
acquiring more data points. Finally, the potential of spICPMS for environmental detection of nanoparticles is demonstrated for a
wastewater treatment plant effluent sample.

T he increasing application of engineered nanoparticles

(ENP) in consumer products is forecasted to have a major
impact on most segments of the future society, and ENP are
currently being used in a large variety of consumer products.1
Following indications that they are potentially harmful for
organisms in the environment,2 the effect and exposure
assessment of ENP need to be supported by in situ analysis
and physicochemical characterization, which is difficult with
currently available methods. For instance, models predicting
the environmental concentrations of ENP in different compart-
ments such as surface waters, soils, and sediments3,4 need to be
validated with environmental measurements,5 but the quanti-
fication of ENP in complex environmental matrixes is very
challenging due to the likely low environmental concen-
trations3,4 (<106 particles mL−1; < pg L−1), the high abundance
of natural nanoparticles,6 and the presence of interfering
elements.7 Similar challenges also apply to cosmetics, food, and
nanomedicine protection goals. To address these challenges,
new sensitive and selective methods working on a single
particle level need to be developed.7−9 Moreover, given many
indications that the mode of action depends on the size of
ENP, these novel methods need to be able to at least quantify
both particle numbers and size distributions. Electron
microscopy methods have been shown to provide powerful
information,10,11 but sample preparation artifacts, the high
particle count required, and unsuitability for high-throughput
applications limit the applicability of these techniques for
environmental samples.
The capabilities of single particle inductively coupled plasma-
mass spectrometry (spICPMS) are explored in this study as a
technique to simultaneously determine size and ultratrace
concentrations of ENP in aqueous samples. If a sufficiently
dilute suspension of metal-rich particles is nebulized into the
plasma, then a burst of ions will be generated when each
discrete particle is vaporized, atomized, and ionized. The ions
of the mass of interest are subsequently counted with a very
high frequency similar to the time scale of a particle event. The
theory of spICPMS for liquid analysis was outlined by
Degueldre et al.,12−16 where they showed that the concen-
tration of particles can be derived from the time-resolved
signals. They also experimentally demonstrated the feasibility of
the method for a range of particles of different materials with
sizes larger than 80 nm.12−16 The spICPMS approach is further
explored in this study for the case of Ag nanoparticles (AgNP).
AgNP can be found in many consumer products today,
particularly because of their antibacterial properties, and
concerns have been raised about emissions into the environ-
ment.17−21 spICPMS has already been applied for character-
ization of the effluent of a washing machine in which AgNP are
used.22 Laborda et al.23 estimated both AgNP particle number
concentration and dissolved silver concentration from
spICPMS measurements.
A 3*σ threshold limit is usually adopted to discriminate the
ion bursts of a particle from the background signal. While 3*σ
is a commonly used definition for detection limits (DL), this
concept cannot be used for spICPMS where the intensity of
particle signals are outliers within a data set of which the mean
Received: December 5, 2011
Accepted: April 6, 2012
Published: April 6, 2012

© 2012 American Chemical Society 3965 dx.doi.org/10.1021/ac203005r | Anal. Chem. 2012, 84, 3965−3972
Analytical Chemistry
is mainly determined by the continuous signal originating from
dissolved analytes. Moreover, it can be calculated that 0.13% of
a Gaussian distributed dissolved signal is erroneously counted
as particles if a 3*σ detection limit is assumed. A Poisson
distribution is probably more appropriate for mean dissolved
signals approaching zero,23 and in this case, ca. 0.5% of the data
points are erroneously counted as particles. Previous work on
spICPMS12−16,23 did not elucidate how many particles need to
be counted in order to reach a certain level of confidence in the
results. In addition, no method for determining the particle size
without the use of standard particles of known sizes, which are
currently not available for most nanoparticles, has hitherto been
presented. In addition, the previous studies by Degueldre et
al.12−16 only showed detection of particles of 80 nm and above.
More recent work showed that 40 nm AgNP can be
discriminated from the dissolved signal,23 but enhanced
reactivities and toxicity have been demonstrated for sizes
below approximately 30 nm.24,25 Consequently, there is a need
to further develop and optimize spICPMS to reduce the lowest
detectable size limit and to improve the methods for
discriminating between the particle events and the background
signal.
The current study investigates the detection capabilities for
AgNP using a sector-field ICPMS (Thermo-Finnigan Element
2). The software of this instrument allows one to reduce dwell
times (also called sampling time), i.e., the detector acquisition
time for each data point below 5 ms and the minimum dwell
time used in previous work.12−16,23 It is discussed how this
affects the size detection limits of spICPMS. A method for
determining particle size using calibration with dissolved
standards is also presented, and data analysis is improved. In
addition, the number concentration detection limit of spICPMS
and how it may be improved is discussed. Finally, as a feasibility
check, an environmental sample from a wastewater treatment
plant (WWTP) effluent is analyzed.
■
THEORY
Signals of Dissolved and Particle Bound Analytes. The
number of ions of dissolved analytes, Idiss, arriving at the
detector in each data point (dwell) during the dwell time, tdwell
(s), is given by:23
Idiss = tdwellfneb cdissAq × fis fion fplasma ftrans
(1)
Equation 1 contains two parts that account for dilution and
efficiency factors in the sample introduction system, and plasma
and mass spectrometer, respectively. In the first part giving the
number of analyte atoms introduced to the plasma, f neb is the
fraction of the analyte that passes through the nebulizer and
spray chamber, the nebulization efficiency, cdiss, is the
concentration of ions in the sample in mol L−1, A is Avogadro’s
number, and q is the flow rate of sample into the nebulizer in
Ls−1. The second part is the counting yield that determines the
fraction of analyte atoms entering the plasma that are detected.
f is is the abundance of the monitored isotope, f ion is the fraction
of analyte that becomes singly charged positive ions in the
plasma, f plasma is the fraction of analyte exiting the plasma
through the sampler cone, and f trans is the transport efficiency or
the fraction of ions that is transported from the sampler cone to
the detector. The terms in the second part of eq 1 are most
likely equal for dissolved or particulate signals because several
studies have shown that there is no significant difference
between conventional ICPMS analysis of acid digested or
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nondigested Au and Ag nanoparticles smaller than 100 nm
diameter.26−28 However, it is possible that the f neb for particles
still differs slightly from that of the dissolved analyte in the
lower concentrations used in this study due to increased
adherence on, e.g., tubing and spray chamber walls. If this is the
case, it would influence number concentration measurements
based on nebulization efficiency (see below), but the analyte
mass per particle reaching the plasma is not affected by f neb.
Ipart, the signal intensity caused by one nanoparticle, can thus be
calculated replacing the first part of eq 1 by the number of
atoms in one particle npart. It should thus also be possible to
measure the mass of analyte in the particles using calibration
with dissolved standards.
Distinguishing Particle Events and Size Detection
Limits. Counting particle events in this study is based on
detecting outliers in the more continuous dissolved signal. In
the ideal case, the signal intensities of dissolved ions do not
overlap with nanoparticle signal intensities in a frequency−
intensity diagram. However, reducing particle size may lead to
overlap with the dissolved signal, due to the cubic relation of
particle size and mass. Additionally, all noise factors of the
dissolved and/or particulate signal increase overlap, e.g.,
increasing number of incompletely measured particle events
or low stability in the dissolved signal. Concentration detection
limits in conventional ICPMS are usually stated in terms of
standard deviation of the background signal, σb. The size
detection limit in spICPMS may similarly be expressed in terms
of standard deviation of the dissolved signal σdiss, but it is not
possible to calculate σdiss unless one can distinguish the
dissolved signal from the particle events. Mitrano et al.26
proposed an iterative algorithm based on a detection limit 3*σ,
where σ is the standard deviation of the whole data set. The
data points exceeding μ + 3*σ, where μ is the mean of the data
set, are considered nanoparticle signals and are removed.
Calculation of μ + 3*σ is then repeated multiple times, and
nanoparticle signals are removed from the remaining data
points until no further data points are removed. More details
can be found in Supporting Information. The resulting
detection threshold signal intensity for particle detection set
by this algorithm is for a low frequency of particle events
approximately Idiss + n*σdiss. This sets the requirement Ipart >
n*σdiss for a particle event to be recognized as such (derivation
in Supporting Information). The choice of detection threshold,
n*σ, in the algorithm is thus critical for the analysis, and n will
be varied in this study.
Counting Particle Events and the Particle Number
Concentration Detection Limit. The average number of
particles λ entering the plasma during a dwell, based on the first
part of eq 1, is given by:
λ = qfneb c ptdwell (2)
where cp is the particle number concentration in the sample.
The probability, pk, for obtaining single (k = 1) and multiple (k
> 1) particle events and the fraction of dwells containing
particle events, pp, can be calculated using Poisson statistics
(further discussion in Supporting Information). The total
number of particle events counted in a sequence of D dwells is
D*pp. At low λ, the chance for multiple particle events, pk>1, is
negligible relative to pk=1 and the approximation pp ∼ λ is valid.
The particle count at the detector increases linearly with cp in
this case but, at higher λ, i.e., higher cp, nebulization efficiency,
flow rate, and/or dwell time, a lower particle count than
predicted from the initial linear trend is obtained. The particle
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Analytical Chemistry
Figure 1. Time resolved signals of spICPMS and the corresponding signal intensity frequency histograms for samples of ultrapure water, a 1 μg L−1
dissolved Ag standard, a fresh 40 nm AgNP suspension, and an aged 40 nm AgNP suspension. The dashed lines indicate the 5*σ detection threshold.
number concentration can be determined either on the basis of
measured nebulization efficiency and using eq 2 or from
calibration curves prepared from particle standards (see below).
The particle number detection limit can be related to the
relative error in particle counting Er (derivation in Supporting
Information):
1
Er = 2z
N (3)
where z is the z-score of the normal distribution which assumes
the value of 1.96 for a 95% confidence interval.29 Given a
desired accuracy, the particle number detection limit thus
depends only on the number of particles counted, which
increases with D.
■ EXPERIMENTAL SECTION
Chemicals. AgNP suspensions were obtained from British
biocell international (BBI, UK) (20, 40, 60, and 80 nm in
nominal diameter). The particles were citrate-coated and
consisted of elemental silver. Unless otherwise stated, the
samples were measured briefly after the first opening of the
container. Reference Au nanoparticle suspensions (30 and 60
nm nominal diameter) were obtained from NIST. Dissolved
silver standards (ultra scientific) for ICPMS calibration were
diluted in ultrapure water from a 1000 ppm solution in 2%
HNO3.
AgNP spICPMS Measurements. A Thermo Element 2
ICPMS (settings given in Table SI-2, Supporting Information)
was used in all experiments after sensitivity and short-term
stability optimization using 1 μg L−1 In in 2% HNO3. A
discussion regarding issues with the dwell times that are specific
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for this instrument is found in Supporting Information.
Dissolved standards were included in each analysis sequence,
and the nebulization efficiency was measured in triplicate using
the waste collection method.30,31 A detailed description of the
measurement of nebulization efficiency and calibration of
particle size using dissolved standards is found in Supporting
Information. The nanoparticle stock suspensions were diluted
in series up to 109 times in ultrapure water. The concentrations
are listed in Table SI-2, Supporting Information. The nominal
dwell time was set at 1, 5, or 10 ms, and 10 000 or 40 000
dwells of 107Ag measurements were collected for each dilution.
Calibration curves, frequency histograms, and particle counts
were calculated from the exported raw data using spICPMS
theory in MATLAB (version 6). For each dilution series, the
particle concentrations were determined using eq 2 and the
measured nebulization efficiency on a sample with a high
enough concentration to obtain sufficient counting statistics
(details in Table SI-2, Supporting Information). These
measured values were used to calculate the concentrations of
the other samples in the dilution series using the dilution factor.
WWTP Samples. WWTP effluents were sampled at the
GRYAAB WWTP (Gothenburg, Sweden) the 19th of May
2009 and analyzed the same day. The sample was a daily flow
composite sample from the final treated water from which three
subsamples were filtered through an acid rinsed and neutralized
5.0 μm Millipore Durapore Millex (PVDF syringe filter with 3.9
cm2 filter area) followed by a 0.45 μm Millipore Durapore
Sterivex-HV (PVDF filter with 10 cm2 filter area). The first 2
mL were discarded, and 10 mL of filtrate was collected into acid
washed and neutralized polycarbonate vials. The dwell time was
not optimized for these samples, and the minimum possible
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dwell time, 0.1 ms, was used together with a relatively large
number of dwells (D = 65 500) to maximize the chance of
detecting a significant number of nanoparticle events in this
first feasibility study that did not involve an elaborate sample
characterization. To calibrate for particle number concentration
dilution series of the NIST, Au standard particles were
measured (calibration curves in Figure SI-8, Supporting
Information). Concentrations in the WWTP samples were
obtained from the calibration curves containing the frequency
of particle events plotted against the number concentration
provided by the manufacturer. To ensure that sensitivity was
not size-dependent to any significant extent, the calibration
curves were measured for both 30 and 60 nm Au nanoparticle
sizes.
■ RESULTS AND DISCUSSION
Dissolved and Particulate Signals. Typical spICPMS raw
data and frequency histograms using 5 ms dwell time are shown
in Figure 1. The low intensity dissolved signal and the particle
related spike in ultrapure water is due to both impurities and
carry-over from previous samples. The signal of the 1 μg L−1
dissolved Ag standard has a RSD of 34% and is relatively
symmetrically distributed around a mean close to 100 ion
counts. Even in the “fresh” 40 nm AgNP sample (analyzed
within one week after delivery), one can recognize a nonzero
dissolved signal resembling that of ultrapure water and particle
events that appear as intense spikes. The intensities of these
particle spikes are lower for the “aged” particle suspension
(analyzed 3 months after opening) compared to the fresh
suspension, suggesting a particle size decrease while the
dissolved signal intensity is increased, probably due to partial
dissolution upon exposure of the fresh standard to oxygen. The
overlap of nanoparticle and dissolved signal is more
pronounced in the aged standard. Figure 1 also shows that a
5*σ threshold value indeed identifies most measured intensities
of ultrapure water and 1 μg L−1 dissolved Ag as below the
particle detection threshold, whereas the nanoparticle events of
the fresh 40 nm AgNP suspension are identified as outliers and
are thus retained as nanoparticle events. This approach also
demonstrates the difficulty in distinguishing nanoparticles from
dissolved signals in the case of overlapping signals as is the case
for the aged 40 nm AgNP suspension. Both the fresh and aged
40 nm standard may contain false positives, i.e., signal
intensities of dissolved Ag that are above the 5*σ threshold
and that are thus erroneously regarded as particle signals, but
the proportion of false positives to total detected particle events
is likely higher in the aged sample than in the fresh sample.
Analogous data for 1 ms dwell time is shown in Figure SI-3,
Supporting Information.
Effect of Dwell Time. Figure 2 shows frequency−intensity
histograms for 80 nm Ag nanoparticles measured using 1, 5,
and 10 ms dwell times. While the intensity of the dissolved
signal increases with tdwell, the particle signal does not. The
overlap between dissolved and nanoparticle signals thus reduces
as tdwell is reduced (equations found in Supporting Informa-
tion). However, the standard deviation of the dissolved signal
and the probability that a particle event is only partially
measured increase with reducing tdwell, especially when the
dwell time is comparable to or lower than the duration of
particle event, (ca. 0.1−0.5 ms).32 Shortening tdwell can thus also
increase the overlap of dissolved and nanoparticle signals, and
an optimum tdwell with a minimal signal overlap therefore exists.
Figure 2 shows that 5 ms is in most cases an optimum dwell
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Figure 2. Frequency−intensity histograms of 80 nm AgNP measured
with dwell times of 1, 5, and 10 ms. The dashed lines indicate the
particle detection threshold.
time because increasing tdwell to 10 ms does not increase
resolution of the particle signal from the dissolved signal. The
broad tail in this sample can be explained by multiple particle
events. The overlap in the data with tdwell = 1 ms is high because
of a large proportion of incompletely measured particle events.
Similar trends were observed for 40 and 60 nm particles
(Figures SI-4 and SI-5, Supporting Information).
Measurement of Particle Size. The particle sizes were
determined by calibrating the ion count with respect to mass of
Ag introduced into the plasma using dissolved standards.
Provided that the chemical composition is known, this allows
estimation of the particle size distribution by assuming a
spherical shape for all particles. The calculated diameters are
thus corresponding volumetric spherical diameters. Figure 3
shows the calculated size distributions of 20, 40, 60, and 80 nm
particles calculated from data obtained using a 5 ms dwell time.
The calculated diameters of the 40, 60, and 80 nm AgNP
standards are in relatively close agreement with the nominal
diameters. Diameter calculation requires the nebulization
efficiency, a value that was determined in this study using an
indirect determination of which the accuracy is disputed,
especially when the nebulization efficiency is relatively low (i.e.,
< 5%).30 However, it can be shown (Supporting Information)
that any error in nebulization efficiency is reduced to its third
root when propagated to an error in diameter. Figure 3 shows
that a 50% variation of the nebulization efficiency results in ca.
± 10 nm variation of the peak maximum of 80 nm particles.
The indirectly measured nebulization efficiencies were 15−20%
where the discrepancy with, e.g., directly measured nebulization
efficiency, a value that is generally regarded as more accurate, is
likely lower than 50%.30 Moreover, direct methods cannot be
run at the same time as the spICPMS determination, whereas
the nebulization efficiency is highly dependent on sample
introduction settings33 that may differ from the conditions
during a direct nebulization efficiency determination where the
plasma is not ignited. However, the accuracy of the nominal
particle sizes was not investigated in this study that was not
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Analytical Chemistry
Figure 3. The size distributions of the 20, 40, 60, and 80 nm nominal
diameter Ag particles determined by spICP-MS. The error bars
demonstrate the effect of possible 50% errors on the determination of
nebulization efficiency on the calculation of size. A tdwell of 5 ms was
used except for the 20 nm AgNP for which it was 1 ms.
aimed at demonstrating the accuracy of particle size
determination of spICPMS.
Since spICPMS measures the mass of particles that varies
with the third power of the diameter, this technique can
potentially achieve a very high resolution in diameter. However,
studies on single droplets introduced into plasma indicate that,
e.g., variation in the point of vaporization and trajectories that
lie off the sampler cone axis may introduce variation in the
intensity of particle signals.34−36 Since there is no information
about the polydispersity of the samples, the size resolution
capabilities could not be assessed in this study. As for particle
sizing by microscopy, a large number of particles are required
for the estimation of the mean diameter with sufficient
accuracy, and the sizing capabilities will ultimately be
determined by the available sample volume.
The distributions of particle event signals of the 20 nm
particles were to a large extent overlapping with the dissolved
signal although a dwell time of 1 ms was used. Only the high
intensity tail corresponding to 20−25 nm volume equivalent
spherical diameters could be discriminated. Figure 3 thus
confirms that 20 nm AgNP are possible to detect and to some
extent possible to discriminate. The size detection limit is for
these samples set by the dissolved background and its standard
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deviation rather than the sensitivity of the ICPMS. Further
improvement of the detection capabilities with respect to size
should thus focus on reducing the dissolved signal intensity and
its standard deviation. Since the samples are diluted up to 109
times to avoid measuring multiple particle events, the dissolved
background predominantly consists of sample carry-over and
not dissolved silver coming from the sample itself. However, if a
detectable concentration of dissolved silver from the sample
remains, the magnitude of its signal can be reduced by
decreasing the sample uptake rate. Reducing the noise of the
dissolved signal can be achieved by, e.g., monodisperse droplet
delivery32,37 or a Peltier cooled desolvation system37 by which
it is possible to partially resolve 20 nm AgNP from the
dissolved background, whereas this is not possible with
conventional sample introduction.37 Developing algorithms
capable of deconvoluting overlapping signals can also
contribute to detection capabilities. The theoretical smallest
detectable equivalent spherical diameter is 6.1 nm, because this
value corresponds to the approximate minimum silver mass
needed to deliver at least one 107Ag ion to the detector; given
that knowing the mass of silver introduced to the plasma during
dwell, it can be calculated that only ca. 1/30 000 of the 107Ag
atoms reaches the detector in a typical measurement.
Choice of Detection Threshold. The number of particles
counted per 10 000 dwells are plotted as a function of the
parameter n for 20 and 40 nm AgNP and for a 2 μg L−1
dissolved standard using a 5 ms dwell time (Figure 4; data for 1
Figure 4. The number of particle events counted by the iterative
algorithm as a function of threshold value (n) for data obtained using a
5 ms dwell time. The following data sets are included: 2 μg L−1
dissolved Ag and 40 and 20 nm AgNP suspensions.
ms dwell time is shown in Figure SI-6, Supporting
Information), suggesting that an n value of at least 4 must be
used to reduce the number of false positives to less than 0.1% of
the total count in the case of the dissolved standard. The
particle count should thus reach a value that is independent of n
if n > 4, in the absence of false positives. This behavior is
approached for the 40 nm particles measured at 5 ms dwell
time where the particle events are completely resolved from the
dissolved signal (Figure 3). In the case of 40 nm particles
measured at tdwell = 1 ms and for all 20 nm data, the particle
events are not resolved from the background and it can be seen
that the particle count continues to decrease upon increasing n.
Increasingly more true particle counts are removed together
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Analytical Chemistry
with false positives in this case because of the overlap of particle
and dissolved signals. It is evident that to reduce the number of
false positives to a predefined acceptable level, e.g., 0.1% of the
total count, n values considerably higher than 3 must be used to
reach this level over a broad concentration range. However,
when very high values for n are used, particles are omitted from
counting. For the remainder of the discussion, n = 5 was
therefore chosen as a compromise between retaining a
sufficiently high particle count over a broad concentration
range and ensuring that contribution of false positives is less
than 0.1% of the actual particle events. (Particle count over a
range of concentrations are compared for n values of 3, 5, and 8
in Figure SI-7, Supporting Information.)
Concentration Calibration Curves. The logarithm of
particle count divided by concentration is plotted as a function
of the logarithm of concentration for the 20, 40, 60, and 80 nm
AgNP measured using 1 and 5 ms dwell time in Figure 5. The
Figure 5. The logarithm of counted particle events divided by the
concentration as a function of the logarithm of concentration. The
data obtained for 20, 40, 60, and 80 nm Ag particles using 1 ms (left)
and 5 ms (right) dwell time and 5*σ detection threshold is shown. 10
000 dwells were used for each sample. The boxes indicate the region
where the relative frequency of false positives (solid line) and multiple
particle events (dashed line) becomes significant.
particles were counted with a 5*σ detection threshold. The data
points in such plots are on horizontal lines in the particle
concentration range where true particle counts dominate over
false positives and a linear response of the experimental particle
count therefore occurs. The level of these lines depends on the
measured particle number concentration, whose accuracy
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depends on the determination of nebulization efficiency, and
the ability to discriminate particle events from the dissolved
signal. Although the number concentrations were not
confirmed in this study, Figure 5 demonstrates the dynamic
range and linear concentration dependent response of
spICPMS particle counting. The contribution of false positives
is significant when the particle concentration is low and the
particle number is consequently overestimated. The frequency
of false positives thus sets the limit of detection with respect to
concentration when present. At higher concentrations, the
counting yield decreases due to the occurrence of multiple
particle events that are each counted as one particle event and a
downward slope can be observed. This behavior is observed
regardless of size or dwell time for the particles investigated
here. A plateau corresponding to the linear response to
concentration is obtained over ca. 2 orders of magnitude of
particle number concentrations, even if particle events are not
resolved from the background, as is the case for 20 nm AgNP
and measurements using 1 ms dwell time. This suggests that
the iterative algorithm may also be successful if the nanoparticle
and dissolved signal intensities partly overlap. However, the
particle count is less accurate in this case because a fraction of
the true particle events are not counted as such. Moreover, any
error in nebulization efficiency is also propagated to the particle
count, and the effective dwell time in eq 2 is not necessarily
equal to the nominal tdwell (further discussion and illustration in
Figure SI-1, Supporting Information).38
Effect of Number of Dwells. Figure 6 shows how the
linear range of particle number determination depends on
Figure 6. The logarithm of counted particle events divided by the
concentration as a function of the logarithm of concentration for 40
nm AgNP measured using a 5 ms dwell and 5*σ detection threshold.
The number of data points used was varied by randomly selecting
from the 40 000 measured data points for each sample.
randomly selecting smaller data point sets among the 40 000
measured dwells. When D = 1000, only one or no particles are
counted for the first 7 data points and the true particle number
is increasingly overestimated at lower concentrations, because
the number of false positives becomes more significant relative
to the total count. Moreover, there is more scattering in data
obtained from smaller data sets, because of higher statistical
fluctuations. Increasing D to 5000 reduces this scatter and a
linear response is obtained over at least 2 orders of magnitude,
but the scatter in data is still significantly higher than for D >
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10 000. Figure 6 thus shows that the concentration detection
limit can be decreased by increasing D. However, the extent by
which this limit is reduced is lower at higher D values as Figure
6 shows that increasing D from 20 000 to 40 000 provides
much less reduction in scatter and the proportion of false
positives. The results agree with eq 3.
Wastewater Effluent Samples. Of the screened elements
(Ag, Ce, Ti, Si, Zn, Cr, Cu, Mo, Pt, Sb, W, Y, and Zr), only the
Ag, Ce, and Ti analyses demonstrated particle events (Figure
7). The filtration through the 450 nm nominal cutoff filters sets
Figure 7. Time resolved signals of spICPMS for wastewater effluent
samples. The monitored elements were Ti, Ce, and Ag.
an upper limit on the particle sizes. Gold particle reference
materials were used to estimate the particle number
concentrations (Figure SI-8, Supporting Information). The
dwell time of 0.1 ms does not allow accurate sizing of the
particles. The numbers of particle events were low and
therefore a 8*σ detection threshold was chosen to obtain a
sufficiently low frequency of false positives. The concentrations
were 9568 particles mL−1 for Ag, 2312 particles mL−1 for Ce,
and 32 656 particles mL−1 for Ti. The predicted environmental
nano-Ag concentration in wastewater effluents for nano-Ag in
Europe is 42 ng L−1.39 Assuming a spherical particle size of 20,
40, and 60 nm, this predicted concentration corresponds to 95
493, 11 937, 354 particles mL−1, suggesting that the measured
particle number concentration in WWTP is in the order of
magnitude of the predicted concentration. It is unknown to
what extent AgNP are being emitted as particles or whether
they in fact occur as dissolved ions in applications such as food
packaging plastics, kitchen utensils and surfaces, textiles,
washing processes, etc., but recent investigations suggest that
both silver forms are reprecipitated as nanoscale silver sulfide
(Ag2S) particles in wastewater treatment plants.40−42 spICPMS
analysis does not allow one to verify whether the detected silver
particles are elemental silver nanoparticles or silver sulfide,
which in this case would also impede size estimation.
Manufactured CeO2 nanoparticles are used in a number of
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applications, e.g., additive to diesel fuel to improve combustion
efficiency, support in the automobile catalysts, and as chemical-
mechanical polishing in certain applications, e.g., electronics
industry. The Gothenburg wastewater system receives road
runoff and industrial and domestic wastewater, so the Ce
particles may originate from man-made sources. TiO2 nano-
particles are used in a large number of applications from
sunscreens and other cosmetics to paint.43 The predicted
environmental concentration in wastewater effluents for TiO2
in Europe is 3.5 μgL−139 leading to 1 975 300, 246 920, and 73
160 particles mL−1, assuming spherical 20, 40, or 60 nm sizes,
respectively; values that are again comparable to the measured
32 656 mL−1, considering the large uncertainty on modeled
values. However, natural sources cannot be ruled out for both
Ce- and Ti-containing nanoparticles. Both brookite TiO2 and
Ce bearing allanite, for instance, have been found in river
sediments.11
■ CONCLUSIONS AND OUTLOOK
AgNP spICPMS signal intensities can be resolved from
dissolved signal intensities when the size exceeds ca. 20 nm
and the dwell time exceeds 1 ms. When these signal intensities
are not fully resolved, the particle number determination is less
accurate. Further improvement of the detection limit can be
achieved by reducing the dissolved signal intensity and its noise,
e.g., using monodisperse drop delivery.32 The choice of a
proper detection threshold is important for the quantification
of the particle number concentration. Using n values of ca. 5
compromises between a relatively low frequency of false
positives and a relatively high effective particle count. The
lowest detectable concentration is determined by the number
of acquired dwells but also by the relative frequency of false
positives. The volume equivalent spherical diameters calculated
from the signal intensities using calibration with dissolved
standards agree with the nominal diameters of the particles,
suggesting that spICPMS can be used as a fast and sensitive
particle sizing method for environmental samples.
The detection of particles in an environmental sample was
demonstrated. The shortest possible tdwell of 0.1 ms was used in
order to minimize the size detection limit. Increasing the tdwell
increases sensitivity with respect to concentration and allows
more accurate determination of the mass of analyzed element
in the particle and thus its size. With the instrumentation used
here, it is only possible to measure a single mass within the
duration of a particle event which does not allow determination
of the composition of individual particles. This limits the
applicability of the method for unknown samples, where it
would be interesting to adapt the concepts developed in this
paper for instruments capable of simultaneous multielement
detection.44 Such a method could, for instance, detect whether
the detected particles are in fact AgNP or whether these are
transformed into Ag2S nanoparticles.
■
ASSOCIATED CONTENT
*
S Supporting Information
Additional information as noted in the text. This material is
available free of charge via the Internet at http://pubs.acs.org.
■
AUTHOR INFORMATION
Corresponding Author
*E-mail: martin.hassellov@chem.gu.se. Tel.: +46-31-786 9050.
Fax: +46-31-77227856.
dx.doi.org/10.1021/ac203005r | Anal. Chem. 2012, 84, 3965−3972
Analytical Chemistry Article

Notes (26) Mitrano, D. M.; Lesher, E. K.; Bednar, A.; Monserud, J.;
The authors declare no competing financial interest. Higgins, C. P.; Ranville, J. F. Environ. Toxicol. Chem. 2011, 31, 115−

■
ACKNOWLEDGMENTS
We thank the following funding agencies for support: CEFIC
Long range research initiative, European Commission FP7
projects NANOFATE and MARINA, the Swedish Environ-
mental Research Council FORMAS, and the Gothenburg
graduate school Environment and Health. We thank Dr.
Nicklas Paxeus at GRYAAB for help with the wastewater
sampling.
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