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Automated standard dilution analysis

Cite this: J. Anal. At. Spectrom., 2020, Willis B. Jones, a George L. Donati, a
Clifton P. Calloway Jrb
35, 178 and Bradley T. Jones*a

Standard dilution analysis (SDA) is a recently described calibration method that combines internal
standardization and standard additions to improve accuracy and precision. By combining these two
Published on 25 November 2019. Downloaded on 1/3/2020 7:14:36 AM.

Received 9th October 2019
Accepted 19th November 2019
DOI: 10.1039/c9ja00339h
rsc.li/jaas
traditional methods, SDA offers corrections for sample matrix effects while also minimizing signal
fluctuations due to changes in the excitation source and the sampling process. In its most basic form,
SDA is performed by introducing a constant amount of sample into an instrument while varying the
amount of standard solution added to the sample. The procedure is simple to perform by hand, but
automation of the process is challenging because the dilution of the standard solution by a blank occurs
on-line while the measurement is taking place. The experiments presented here are aimed at improving
the automation of the SDA process. Inductively coupled plasma optical emission spectrometry (ICP OES)
is used as a model for SDA applications, although the calibration method is applicable to any technique
that accepts samples as a liquid flow. The end goal of automation is for successive samples to be
analyzed quickly by using an automatic sampler to switch from sample to sample, as it is the case in
routine analytical methods. The required dilution of the standard by a blank mixture is achieved through
the use of a two-channel pinch valve, with dilution occurring by diffusion as the solution is pulled from
the valve toward the plasma by a second channel in the instrument's peristaltic pump. The proposed
automated SDA method is effective, with analyte recoveries within a few percent of 100 for the elements
evaluated (i.e. Al, Cd, Co, Cr, Cu, Fe, Ni and Pb) and nine different sample matrices. The method also
provides promising results when analyzing real-world samples. The limits of detection obtained with the
automated SDA method are similar to those typically found for external standard calibration and ICP
OES, falling within the single-digit mg L
1 range.

the calibration standards. The SA method is cumbersome and

Introduction
Accurate calibration is the ultimate goal of most analytical
techniques, as it allows for determination of the concentration
of analytes in real-world samples. At its core, calibration
involves the preparation of a series of standard solutions con-
taining analytes of known, increasing concentration, which
allows for the calculation of the unknown analyte concentration
in a sample. The simplest of the traditional calibration
methods, external standard calibration (EC), is performed by
direct comparison of a sample to a series of calibration solu-
tions. It assumes that the sample matrix does not affect the
analytical signal to a signicant degree when compared to an
identical concentration of analyte prepared in pure solvent,
which usually is an inaccurate assumption.1 More advanced
calibration methods have been designed to correct for problems
that arise in simple EC. The standard additions (SA) method
introduces an identical amount of sample solution into each of
time consuming, especially if multiple samples are being
analyzed, but it is effective at correcting for matrix effects, as the
sample matrix is present in each calibration standard in equal
parts.2,3 The internal standard (IS) method is put into practice if
the analytical signal is affected by changes in the sampling
process or the excitation source. An extra species, called the
internal standard, is added to all calibration and sample solu-
tions. For an ideal IS species, changes in the sample position,
volume, or analyte atomization and excitation processes affect
the analyte as well as the internal standard, thus these external
changes are corrected.4 In addition to these traditional cali-
bration methods, other correction strategies such as extrapo-
lation to innite dilution,5 multi-energy calibration,6 and multi-
ow calibration7 have also been proposed in order to correct for
matrix effects.
Standard dilution analysis

a
Standard Dilution Analysis (SDA) is a recently described cali-
Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA.
E-mail: jonesbt@wfu.edu bration strategy that combines the traditional SA and IS
b
Department of Chemistry, Physics and Geology, Winthrop University, Rock Hill, SC methods.8 It is simple to perform, and requires only two solu-
29733, USA tions for a full calibration. The rst solution consists of 50%

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sample and 50% standard. The standard contains both the
analytes and an internal standard. The second solution consists
of 50% sample and 50% blank. SDA is performed as the second
solution is poured into the rst, resulting in an on-line dilution
of the standard while the amount of sample is kept constant.
SDA calculations are also simple, as described in detail below.
This new calibration method has been successfully applied to
multiple analytical techniques, including ame atomic emis-
sion spectrometry (FAES),9 ame atomic absorption spectrom-
etry (FAAS),10 microwave-induced plasma optical emission
spectrometry (MIP OES),11,12 inductively coupled plasma OES
(ICP OES),8,13–15 ICP mass spectrometry (ICP-MS),16 Raman
spectroscopy,17 and visible absorption spectrometry.8
In practice, SDA offers signicantly improved accuracies and
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alternating between standard and blank solutions eliminates
the need for an elongated cleaning step beyond the typical rinse
time adopted between samples. This is possible because the
dilution occurs in a separate channel from the sample intro-
duction itself.
SDA theory
The calculations used in SDA have changed since their original
derivation,13 and the newest, simplest iteration is described
here. Again, SDA functions on the premise that there is
a constant amount of sample present in an analytical system,
while the ratio of standard to blank is varied. Thus, the
measured analyte signal has contributions from both the

precisions when compared to traditional EC and IS methods, sample itself and the calibration standard, as described by eqn
while offering slight improvements over SA.8,13 Two reviews are (1), where S is the signal, A is the analyte, and std and sam
referenced that incorporate SDA into summaries of other correspond to the standard and sample, respectively.
commonly used calibration techniques in atomic spectrom-
etry.18,19 The most signicant benet that SDA provides over Stotal
A ¼ Sstd
A + SA
sam
(1)
traditional SA is its improved sample throughput due to
a simpler sample preparation procedure. In SDA, a full cali- To simplify the math, a constant K is dened in eqn (2) as the
bration is performed with only two solutions. On the other ratio of analyte signal in the standard solution alone to internal
hand, an important challenge for implementing SDA in routine standard signal, where I represents the internal standard. This
analysis is associated with its automation. In its earliest itera- denition of K allows for eqn (1) to be rewritten as shown in eqn
tions, SDA was performed manually, by pouring the blank- (3).
containing solution into the standard-containing solution. SAstd
Recent advances in the method have been aimed at the auto- K¼ (2)
SI
mation of solution mixing, namely the use of a three-port
mixing chamber directly coupled to the sample introduction
Stotal
A ¼ KSI + Ssam
A (3)
system of an ICP OES.13 This setup allowed for SDA to be per-
formed in its entirety using only the commercial instrument's Eqn (3) is the equation for a line when the analyte signal is
own automatic sampler to dilute the standard-containing plotted against the internal standard signal, with a slope of K
solution with the blank. However, it involved an elongated and a y-intercept that is equal to the analyte signal that arises
rinsing step to completely wash out residual sample from the from the sample alone. Now consider the ratio of this intercept
mixing chamber before proceeding to the next cycle. In another and slope, as shown in eqn (4).
work, a multi-channel nebulizer was used to perform SDA
Intercept Ssam SI
inside the spray chamber of an ICP OES instrument. However, ¼ A ¼ SAsam std (4)
automation was also limited in this case, as the instrument's Slope K SA
automatic sampler was not used and the sample had to be fed
manually into the system.15 In general, a measured signal is equal to the concentration of
The method presented here uses a new version of SDA analyte, C, multiplied by the calibration sensitivity, m.
solution preparation in order to speed up the automation Substituting this in to eqn (4), the concentration of the analyte
process. These experiments use ICP OES as an application in the sample is now present in the calculation, as shown in eqn
model for the calibration strategy. However, it is essential to (5).
reiterate that SDA is not limited to ICP OES, and it may be
Intercept SI SI
applied to any technique capable of accepting liquid samples ¼ mA CAsam  ¼ CAsam std (5)
Slope mA CAstd
CA
and simultaneously monitoring multiple analytical signals. As
mentioned previously, SDA requires a constant amount of
sample to be introduced into the analytical system, while the This equation can be rearranged to allow for direct calcula-
amount of standard is varied through dilution with a blank. To tion of the analyte concentration in the sample, as shown in eqn
simplify the automation of the method, the two calibration (6).
solutions are introduced into the ICP using two separate
intercept CAstd
channels on the instrument's peristaltic pump. One channel is CAsam ¼ (6)
slope SImax
connected to the instrument's automatic sampler, which moves
between individual samples as would be the case in any typical
analysis. The second channel introduces the standard and When the analyte in the standard is undiluted, the internal
blank solutions in an alternating fashion. Consistently standard must also be undiluted, thus the internal standard

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signal is at its maximum. The maximum internal standard
signal can be estimated by rst introducing the standard into
the experimental system and allowing the signal to stabilize
before dilution by the blank,8 or it can be calculated by intro-
ducing a second internal standard into the blank solution,
providing an inverse relationship between the two internal
standards. The internal standard present with the analytes in
the standard solution is referred to as the primary internal
standard, while the internal standard present in the blank is
referred to as the secondary internal standard. The two internal
standards are plotted against each other, and the maximum
internal standard signal is dened as the y-intercept of this
second plot. The use of a secondary internal standard improves
sample throughput, as the maximum primary internal standard
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signal is calculated from the same points in time that provide
the full calibration.13
Note that the nal calculation presented in eqn (6) assumes
that the sample solution and the standard plus blank solution
mix are present in the analytical system in equal parts. The
addition of a simple dilution factor to eqn (6) corrects for an
experiment in which the sample to standard plus blank ratio is
not one to one.
Experimental
Solution preparation
Standard and blank solutions were prepared using distilled-
deionized water (DDI) and trace-metal grade nitric acid. A
multi-element stock solution containing Al, Cd, Co, Cr, Cu, Fe,
Ni and Pb as analytes was prepared using 1000 mg L
1 single-
element standards. This solution was prepared in such a way
that the selected analyte lines provided intensities on the same
order of magnitude. Scandium and Y were used as internal
standards, also prepared from 1000 mg L
1 single-element
standards. The stock solution was diluted in 1% v/v HNO3,
and contained nal concentrations of 1.36 mg L
1 Al and Cu,
2.27 mg L
1 Cr, 4.54 mg L
1 Cd, Co, Fe and Ni, and 13.6 mg L
1
Pb as analytes, with 0.5 mg L
1 Y as an internal standard. The
blank solution, prepared in 1% v/v HNO3, contained 0.5 mg L
1
Sc as the second internal standard and 100 mg L
1 Li to help
balance the ionic strength of the two solutions.
Nine different sample matrices were spiked with the stock
solution to have the same added analyte concentration as the
diluted standard, in order to determine how well the automated
SDA process corrected for matrix effects. The examined
matrices included DDI, tap water, 50% v/v ACS grade nitric acid,
40% v/v ethanol, a so drink, a zero-sugar so drink, a red wine,
50% v/v mouthwash, and 1% m/v Ca.
Several reference materials were analyzed in order to eval-
uate the accuracy of the automated SDA method. A water
pollution standard (VHG Labs, Manchester, NH, USA) was
diluted in 1% v/v HNO3 before performing SDA. A standard
reference material of Oyster Tissue (NIST 1566B) from the
National Institute of Standards and Technology (NIST, Gai-
thersburg, MD, USA) and two standards of metals in hydro-
carbon oil (300 and 900 mg g
1, Agilent Technologies,
Wilmington, DE, USA) were subjected to microwave-assisted
180 | J. Anal. At. Spectrom., 2020, 35, 178–187
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digestion before analysis. The digestion program used with an
Ethos Up system (Milestone, Sorisole, Italy) consisted of
a 15 min ramp to 200  C, a 15 min hold at 200  C, and a 10 min
cooling step. Aliquots of 1.5 mL trace-metal grade HNO3, 6.5 mL
DDI and 2 mL 30% v/v H2O2 were used in the digestion process.
A second stock solution was prepared for the analysis of the
reference materials and for determination of the method's
limits of detection (LODs). A multi-element standard containing
all of the 8 analytes and Sc as an internal standard at 100 mg L
1
(Teledyne Leeman Labs, Hudson, NH, USA) was diluted 1 : 100
in DDI with added 1% v/v HNO3. A second blank was prepared
in 1% v/v HNO3, with 0.5 mg L
1 Y as an internal standard and
100 mg L
1 Li to help balance the ionic strength of the two
solutions.
The analyte and internal standard emission lines used here
are provided in Table 1. Emission intensities of each analyte
and internal standard were monitored at two different wave-
lengths, but only results from one line are presented here. The
use of different emission lines for a given analyte changes the
calculations to a minimal extent, and the use of multiple
internal standard emission lines is even more inconsequential.
Unless otherwise noted, all results discussed here use the
measured intensity of the lines provided in Table 1.
Automation of SDA
The instrument used to evaluate the automated SDA method
was an Agilent 5110 ICP OES (Agilent Technologies, Santa Clara,
CA, USA). However, as previously discussed, it is applicable to
any system that accepts samples as a liquid ow, and is thus not
limited to ICP OES. Fig. 1 provides a basic schematic of how the
SDA process has been automated in this experimental setup.
Solutions are introduced into the ICP through the use of the
standard peristaltic pump and nebulizer on the instrument.
The novelty of the automation technique presented here lies
in how the sample and standard plus blank solutions are
introduced into the plasma simultaneously, with the standard
being diluted from a maximum to blank level in an alternating
fashion. Sample solutions are placed in an automatic sampler
and drawn up through the tubing by the peristaltic pump. A
second channel on the peristaltic pump creates the standard
Table 1 Selected analyte and internal standard emission lines. Species
designation is typical spectroscopic nomenclature, with I signifying an
atomic line, and II designating a singly positively-charged ion
Wavelength
Element Species (nm)
Al I 396.152
Cd I 228.802
Co II 237.863
Cr II 267.716
Cu I 324.754
Fe II 259.940
Ni I 341.476
Pb I 405.781
Sc II 361.383
Y II 371.029
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Fig. 1 Schematic of the automated SDA system. Samples are placed in an automatic sampler. The standard plus blank dilution is introduced
through the use of a two channel pinch valve. The peristaltic pump introduces the sample and standard plus blank mixture into the plasma at the
same rate, as the two solutions travel through identical pieces of tubing. A plastic Y junction is used to allow the pump to draw either standard or
blank through the same piece of tubing, as well as to combine the sample and standard plus blank flows before introduction into the plasma.

plus blank mixture through the use of a two channel pinch valve Table 2 ICP instrumental parameters
(Cole-Parmer, Vernon Hills, IL, USA). Tubing from one channel
Instrumental parameter Operating condition
in the valve draws the prepared standard solution, while the
second channel draws from the prepared blank solution. One Plasma
channel of the valve is open by default. When 12 V are applied to RF applied power 1.20 kW
the valve, the rst channel closes and the second one opens. A Viewing mode Axial
digital timer outlet (NEARPOW, TX, USA) is used to switch a low Nebulizer gas ow rate 0.7 L min
1
Plasma gas ow rate 12.0 L min
1
cost, simple 12 V DC power supply (120 VAC to 12 VDC) adapter, Auxillary gas ow rate 1.0 L min
1
from off to on, and vice versa, every 60 s. This results in an
alternating ow of pure standard or blank solution once per General
minute. The two channels are joined together into a single piece Pump speed 12 rpm
of tubing by a plastic Y junction. The pinch valve allows for the Uptake delay 90 s
Stabilization time 30 s
standard and blank solutions to be interchanged without the
Rinse time 30 s
introduction of air bubbles into the tubing. As the liquid ow Replicates 100
remains constant, the standard solution is diluted by the blank Read time 1s
solution on-line inside of the tubing as it travels from the valve
through the peristaltic pump. The sample and the standard
plus blank ows are joined together on the back end of the default values. Only minimal changes were required to the
peristaltic pump by a second Y junction before introduction soware in order to run automated SDA. The pump speed was
into the nebulizer and into the plasma. kept constant during the rinse step instead of speeding it up in
All instrumental parameters used in the present work are order to preserve the nature of the standard plus blank dilution.
provided in Table 2. All of the ICP OES operating parameters The read time and number of replicates are the two most
were kept at their default values. As the pinch valve setup important parameters when SDA is performed. The read time
provided dilution of the standard plus blank mixture indepen- was kept short (to the soware minimum of 1 s) to provide the
dent of the ICP OES0 automatic sampler, the uptake delay and highest possible time resolution of the signal change provided
stabilization time relate to the introduction of the sample itself by the dilution of the standard by the blank, and vice versa. The
into the plasma. These parameters were also kept at their number of replicates was kept high (to the soware maximum

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of 100) to provide additional points in the SDA calibration curve
(as each point in time provides one point on the curve).
Results and discussion
Basic examples
Representative time-resolved intensity traces for three analytes
and both internal standards are provided in Fig. 2. An addi-
tional standard solution was prepared for the sake of making
this representative plot. The analytes and primary internal
standard, Sc in this example, are present in the standard solu-
tion, and thus signal intensities increase for all of them
following the same prole, at the same time. The secondary
internal standard, Y in this example, is present in the blank
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solution as a means of calculating the undiluted signal of the
primary internal standard. Its signal intensity increased at the
same time as the analytes and primary internal standard
decreased. Note that in the representative traces provided in
Fig. 2, as well as investigations of reference standards and
determination of limits of detection, Sc served as the primary
internal standard while Y was the secondary internal standard
present in the blank. The primary and secondary internal
standards are reversed for the other further discussed examples.
As the valve was switching channels every 60 s, the signal
intensity traces switch from increasing to decreasing every 60 s.
Fig. 2 Time resolved intensities for three analytes (Cd, Cr and Pb) and two internal standards (Y and Sc). The red curve corresponds to the primary
internal standard present in the standard solution. The black curve corresponds to the secondary internal standard present in the blank solution.
The gray boxes represent two successive SDA measurement windows, with the rinse, sample uptake, and stabilization times accounting for the
gap between.
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Although the channels in the pinch valve operate in an on/off
logic fashion, the intensities changed rapidly over a time of
approximately 30 s before stabilizing instead of changing
immediately. This means that the dilution of the standard by the
blank happened by diffusion as the solution travels from the
valve through the peristaltic pump and into the plasma. That
explains why one observes a ramp rather than a jump in the
signal proles, as shown in Fig. 2. The gray boxes in Fig. 2 show
representative times for which a single automated SDA run is
performed (100 time points), with the rinse, sample uptake, and
stabilization times accounting for the gap in between. A 100-
point collection consisted of a full cycle of signal change from
minimum to maximum and back to minimum again, but not
necessarily beginning and ending with a maximum or minimum.
Fig. 3 provides an example of the plots needed to perform
SDA calculations. The sample was DDI spiked with 2.27 mg L
1
Cr, which is the same concentration of Cr present in the
prepared standard. Recall that Y is present in the standard
solution as an internal standard, with Sc in the blank solution
as the secondary internal standard. The Cr versus Y signal plot
has a y-intercept of 16 304 and a slope of 0.437. The Y vs. Sc
signal plot has a y-intercept of 36 988. Eqn (7) provides an
example SDA calculation using the data shown in Fig. 3, which
return a result of a 2.29 mg L
1 Cr, only a + 0.9% error from the
true spike value of 2.27 mg L
1.
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Fig. 3 Example plots needed to complete an SDA calculation, including analyte vs. primary internal standard (left), and primary internal standard
vs. secondary internal standard (right). These two plots and the known concentration of analyte in the standard are all that is needed to calculate
the concentration of analyte in a sample.

intercept CAstd 16304 2:27 mg L
1 Cr samples. The topmost trend in Fig. 4 (black diamonds) corre-
CAsam ¼  max ¼  sponds to spiked DDI, which was the cleanest matrix chosen.
slope SI 0:437 36 988
Note that the other 8 matrices all suppressed both the analyte
¼ 2:29 mg L
1 Cr (7)
and internal standard signals, at least to a slight degree, as
evidenced by the lowering of both the y-intercept and maximum
internal standard signal. This makes it simple to envision that
Robustness of automated SDA an external calibration method with standards prepared in DDI
would provide inaccurate results, as most of the examined
Fig. 4 displays SDA plots obtained for Cr spiked into 9 sample
sample matrices suppress the analytical signal.
matrices. This analyte was not detected in any of the unspiked

Fig. 4 SDA plots obtained for Cr spiked into 9 different matrices. The topmost curve (black diamonds) corresponds to spiked deionized water.

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Note that the slope of the line was preserved between sample
matrices. This is predicted by the theory, as the slope is dened
as the signal ratio of the analyte present in the standard alone to
the internal standard. If this ratio was not preserved between
samples, the internal standard reacts differently to sample
matrices than the analyte, making it a poor choice of internal
standard (in a traditional sense) when examining multiple
sample types. However, SDA would still work if the lines have
different slopes, as the sample matrix for a given sample is
preserved between the standard and blank solutions for an
individual sample run.
Full investigations comparing SDA to traditional calibration
methods have been performed and published previously.8,13 The
focus of this research was simplifying the automation of SDA,
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and thus a comparison with a traditional calibration method
was not performed. Nevertheless, previous results can be
summarized as follows:8,13 SDA provides improved accuracy and
precision when compared to a simple external standard cali-
bration or internal standard method, while providing similar or
slightly improved results compared to the traditional standard
additions method. The major benet provided by SDA over
standard additions is the simplied solution preparation, and
thus improved sample throughput.
Each of the 9 matrices was initially run in triplicate. One
additional measurement was taken for the tap water, soda, and
wine matrices, resulting in a total number of 30 measurements
for each element. This additional measurement was to assess
the effect of small bubbles passing through the peristaltic pump
tubing, as shown in Fig. 5.
Traditional ICP OES methods involve averaging emission
signals over extended lengths of time in order to improve signal
precision. However, this is not possible in the case of automated
SDA, as the signals need to be collected as quickly as possible in
order to achieve a wide range of signal coverage. If too long of
a time period is averaged, the dilution of the standard by the
blank is lost in the averaging, resulting in no signal change over
time. Collecting data points quickly provides many points on
Fig. 5 Influence of bubbles passing through the peristaltic pump tubing. Samples were wine (blue), 1% m/v Ca (red), and tap water (yellow) spiked
with all eight analytes. Both time resolved Cr emission signals (left) and the corresponding Cr SDA plots (right) are provided. The linear fits shown
depict all 100 collected points, including those that fall far from the line due to a bubble passing through the tubing.
184 | J. Anal. At. Spectrom., 2020, 35, 178–187
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individual calibration curves. Although short measurement
times are more subject to noise than longer integrations, the
additional points collected in a single calibration averages the
noise out in the end.
When a bubble comes through the pump tubing, disconti-
nuities appear in the time resolved signals, as shown by the Cr
emission intensities on the le-hand side of Fig. 5. The domi-
nant, slowly increasing and decreasing signals are created by
the dilution of the standard by the blank, and vice versa, that
allow SDA to function. The sharp deviations in signal that occur
over two or three data points were caused by bubbles. SDA plots
of these data sets, bubbles included, are provided on the right-
hand side of Fig. 5. The linear ts depict all 100 data points,
including the bubble points that fall far away from the line. The
sheer number of points in the calibration curve allows for the
method to correct for a bubble or two passing through the
sample introduction line over the course of the measurement.
Recall that the sample matrices were spiked with 2.27 mg L
1
Cr. When including all 100 data points, the data sets shown in
Fig. 5 return a calculated spiked Cr concentration of 2.31, 2.07,
and 2.32 mg L
1 in the wine, 1% m/v Ca, and tap water matrices,
respectively. When removing the points caused by bubbles,
these calculated concentrations are basically unchanged in
wine and 1% m/v Ca, and only slightly changed in tap water,
returning values of 2.30, 2.06, and 2.20 mg L
1, respectively,
showing that this method is capable of handling bubbles
passing through the pump tubing, even when collecting data
points quickly.
Limits of detection
Limits of detection (LODs) were calculated as three times the
standard deviation of the concentration found for each element
in 10 successive replicates of unspiked DDI. The standard used
for LOD determinations was the previously discussed
100 mg L
1 multi-element standard containing all 8 analytes
and Sc as an internal standard. This solution was diluted down
to both 10 and 1 mg L
1 concentrations, and LODs were
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Table 3 Limits of detection obtained using two different standard Table 5 Average percent recoveries obtained for 9 sample matrices
concentrations across 8 spiked elements

LOD (mg L
1) Matrix Recovery (%) n

1
1
Element 1 mg L standard 10 mg L standard DDI 100  2 24
Tap water 102  6 32
Al 0.006 0.05 50% v/v nitric acid 98  2 24
Cd 0.005 0.03 40% v/v ethanol 99  3 24
Co 0.005 0.04 Soda 99  1 32
Cr 0.002 0.04 0 sugar soda 98  4 24
Cu 0.003 0.05 Wine 100  3 32
Fe 0.006 0.05 1% m/v Ca 92  2 24
Ni 0.008 0.05 50% v/v mouthwash 99  2 24
Pb 0.1 0.06
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calculated using both dilutions. The values calculated for each
element are provided in Table 3.
The LODs are largely similar for each element, with lead
being the lone exception. When using a 1 mg L
1 standard, the
LOD for each element (except lead) is on the order of single digit
parts per billion. When using a 10 mg L
1 standard the LODs
increase similarly, up an order of magnitude, to tens of parts per
billion. The LOD for lead is on the order of 100 mg L
1 for both
standard concentrations. These are typical LODs for ICP OES
when using traditional calibration methods. The LODs differ
when using different concentration standards because a full
calibration is collected every time SDA is performed.
When performing a calibration by SDA, using a higher
concentration standard results in more uncertainty in values
near zero. If the standard concentration is the only parameter
that is changed between two SDA calibrations, the total number
of points in the changing signal prole is unchanged. Thus
using lower concentrations improves estimates of the noise
level as the detection limit is approached, as there are more
points in the calibration at low standard concentrations
because the maximum standard concentration is lower. As a full
calibration is required in order to return a blank signal using
SDA, using a lower concentration standard provides slightly
lower LODs.
Spiked sample matrices
Before comparing the results obtained across the spiked
sample matrices, each matrix was run in triplicate without the
spike to determine the amount of analytes inherently present
Table 4 Average percent recoveries obtained for 8 spiked elements
across 9 sample matrices
Element Recovery (%)
Al 98  3
Cd 99  3
Co 98  3
Cr 100  3
Cu 99  7
Fe 100  3
Ni 97  3
Pb 96  3
in each matrix. Most of the analytes returned concentrations
near or below their LODs in every matrix. The exceptions were
approximately 100 mg L
1 Cu in tap water, 120 mg L
1 Cu in
soda, and 700 mg L
1 Al, 200 mg L
1 Cu, and 2 mg L
1 Fe in
wine. The average blank concentrations for each element in
every examined sample matrix were subtracted from their
corresponding spiked matrix results before calculating
recovery values.
Table 4 displays the average percent recovery calculated for
each analyte element across the 9 sample matrices. The auto-
mated SDA method accurately corrects for matrix effects regard-
less of the analyte selected, returning a recovery of near 100% for
each element. Table 5 displays the same data in a different way,
showing the average percent recovery calculated for each sample
matrix across the 8 spiked analytes. Again, SDA is able to provide
accurate results across a wide range of matrix types. Even a 1% m/
v Ca solution, which is a particularly difficult matrix, returns an
accurate average recovery of 92%. The repeatability of the auto-
mated SDA method was examined by running spiked DDI, 1% m/
v Ca, and 50% v/v mouthwash matrices in 10 successive repli-
cates. The results are shown in Table 6. As expected from the
previous results, both DDI and 50% v/v mouthwash return
concentrations within a few percent of the spiked value. The 1%
m/v Ca matrix again returns results that are below the spiked
value, but still accurate especially considering the difficulty of the
matrix itself. Regarding the precision of the automated SDA
technique, the RSDs were on the order of 3% irrespective of the
matrix examined. It is important to note that accuracy and
precision need to be combined in order to provide better esti-
mates of uncertainty between different measurement tech-
niques.20 As the goal of this work was to simplify the automation
of SDA as a calibration technique, a full comparison of SDA to
other traditional calibration methods was not performed, but has
been previously reported. SDA has been shown to offer similar
n results to traditional standard additions methods, while
improving upon simple external calibration or internal stan-
30 dardization methods.8,13
30
30
30 Reference standards
30
30 Several reference materials were analyzed to reemphasize the
30 applicability of automated SDA to real-world samples. Sample
30 digestions and the samples themselves were described in the

This journal is © The Royal Society of Chemistry 2020 J. Anal. At. Spectrom., 2020, 35, 178–187 | 185
 View Article Online

JAAS Paper

Table 6 Repeatability of SDA measurements across three different spiked sample matrices

Spike DDI matrix 1% Ca m/v matrix 50% v/v mouthwash matrix

1
Element mg L Avg StDev RSD (%) Accuracy (%) Avg StDev RSD (%) Accuracy (%) Avg StDev RSD (%) Accuracy (%)

Al 1.36 1.34 0.04 2.7
1.4 1.42 0.09 6.3 4.6 1.44 0.05 3.6 5.9
Cd 4.54 4.5 0.1 2.5
1.4 4.2 0.1 3.0
8.4 4.5 0.1 3.2 0.06
Co 4.54 4.5 0.1 2.6
1.9 4.1 0.1 3.2
9.5 4.4 0.2 3.7
2.1
Cr 2.27 2.27 0.06 2.7
0.1 2.09 0.08 3.7
7.8 2.27 0.09 3.9
0.04
Cu 1.36 1.34 0.03 2.5
1.9 1.25 0.03 2.7
8.3 1.34 0.05 3.4
1.3
Fe 4.54 4.5 0.1 2.6 0.2 4.2 0.2 3.8
7.5 4.5 0.2 3.7 0.07
Ni 4.54 4.4 0.1 2.4
2.6 4.0 0.1 2.9
11.6 4.5 0.2 3.4
1.9
Pb 13.6 13.0 0.3 2.4
4.9 12.1 0.3 2.6
10.9 13.2 0.5 3.7
3.3
Published on 25 November 2019. Downloaded on 1/3/2020 7:14:36 AM.

Table 7 Analyte concentrations and recoveries obtained for SDA determinations by ICP OES while analyzing reference materials

Water pollution standarda Oyster tissueb 300 mg g
1 metals in oilc 900 mg g
1 metals in oilc

Concentration Recovery Concentration Recovery Concentration Recovery Concentration Recovery

Element (mg L
1) (%) n (mg L
1) (%) n (mg L
1) (%) n (mg L
1) (%) n

Al 4.53 91  8 6 0.38 93  3 3 0.40 110  6 6 1.10 103  3 6

Cd 0.27 110  2 6 0.34 94  2 6 0.99 93  2 6
Co 0.97 98  3 6
Cr 1.01 102  3 6 0.25 67  1 6 0.84 79  2 6
Cu 1.12 113  3 6 0.16 107  4 3 0.34 93  2 6 1.00 94  3 6
Fe 0.96 97  5 6 0.41 95.3  0.8 3 0.32 88  2 6 0.90 85  3 6
Ni 1.17 118  4 6 0.37 102  2 6 1.13 106  3 6
Pb 1.08 109  4 6 0.32 89  4 6 1.04 97  5 6
a
VHG Labs, Manchester, NH, USA. b NIST, Gaithersburg, ND, USA. c Agilent Technologies, Wilmington, DE, USA.

solution preparation section. A summary of the obtained results
is provided in Table 7. Blanks in the table correspond to
elements that were not certied in concentration. The concen-
trations reported are those obtained directly from the SDA
calculations, not accounting for dilution or digestion, to allow
for comparison to LODs.
Overall the results obtained using automated SDA for real
samples were accurate, with average recoveries largely falling
between 90 and 110%. The lone exception is Cr in the hydro-
carbon oil samples, which provided recoveries signicantly
below 90%. However, the results for the EC method applied to
the same sample were worse than those obtained by SDA, which
suggests not a problem with the SDA method in the determi-
nation of Cr, but more likely a problem during the sample
preparation.
The results obtained for Al in the water pollution standard
and oil standards used a secondary emission line at
167.019 nm. This was necessary due to an interfering emission
that was observed in the sample causing a falsely high recovery
value.
traditional calibration methods, SDA offers corrections for
sample matrix effects using standard additions while also
offering corrections for changes in the excitation source and
sampling process through the use of an internal standard.
SDA provides results that are superior (or at worst compa-
rable) to simple external standard calibration and internal
standardization methods, while providing slightly improved
results when compared to traditional standard additions
alone.8,13
Experiments presented here were aimed at improving the
automation of the SDA process, allowing for successive samples
to be analyzed quickly by simply using an automatic sampler to
switch from sample to sample, as is the case in most routine
analytical methods. The automation of the dilution of the
standard plus blank mixture was achieved through the use of
a two-channel pinch valve, with the dilution occurring by
diffusion as the solution is pulled from the valve to the plasma
by a second channel in the instrument's peristaltic pump. The
proposed automated SDA method proved to be effective over
various sample matrices, and also provided promising results
when analyzing real-world samples.

Conclusions
Conflicts of interest
Standard dilution analysis has proven to be a robust, simple to
perform, accurate, and precise calibration method as it has The authors declare that there are no conicts of interest
been developed over the past several years. Combining two associated with this manuscript.

186 | J. Anal. At. Spectrom., 2020, 35, 178–187 This journal is © The Royal Society of Chemistry 2020

Paper
Acknowledgements
The authors would like to thank the Graduate School of Arts and
Sciences and Department of Chemistry at Wake Forest Univer-
sity for supporting this study.
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