Accred Qual Assur (2005) 10:190196

DOI 10.1007/s00769-005-0908-4

Yoshihiro Hirano
Kyoko Imai
Kazuo Yasuda

Received: 4 August 2004

Accepted: 11 August 2004
Published online: 21 April 2005

Springer-Verlag 2005
Y. Hirano ())
Hitachi High-Tech
Manufacturing & Service Corporation,
882 Ichige, Hitachinaka,, Ibaraki,
312-8504, Japan
e-mail:
hirano-yoshihiro@naka.hitachi-hitec.com
Tel.: +81-29-2766155
Fax: +81-29-2755278
K. Imai K. Yasuda
Hitachi High-Technologies Corporation,
882 Ichige, Hitachinaka,, Ibaraki,
312-8504, Japan

PRACTITIONERS REPORT

Uncertainty of atomic absorption spectrometer

Abstract The uncertainty of the

flame atomic absorption spectrometer
was studied. Drift of signal, noise,
and nonlinearity of the calibration
curve were found to be main sources
of uncertainty for the absorbance
measurement, and factors affecting
these sources of uncertainty were
evaluated using Cd as a typical element. The combined uncertainty was
0.00133 absorbance units at a Cd
concentration of 0.5 mg/l (0.078 absorbance units). The relationship between the flame conditions and the
burner optics contributed strongly to
the uncertainty. Therefore, the matrix

Introduction
Atomic absorption spectrometry (AAS) has been widely
used for determination of trace metals in various areas,
such as environmental, biological, clinical and material
analyses. The system of the atomic absorption spectrometer is relatively simple. However, there has been no
paper on the estimation of the uncertainty for the instrument.
Several works on the estimation of uncertainty in AAS
have been reported. Kurfrst et al. [1] studied the uncertainty in the analysis of solid materials by graphite
furnace AAS comparing sample digestion AAS and solid
sampling AAS. Stringari et al. [2] evaluated sample
grinding methods for analysis of plant samples by
graphite furnace AAS. Bettencourt da Silva et al. [3]
applied their uncertainty validation method to the analysis
of leaves by acid digestiongraphite furnace AAS. Synek
et al. [4] studied the uncertainties of mercury determination by cold vapor AAS and reported that the contribution of the sample absorbance component was highest.

effect of the sample must to be taken

into consideration in actual analyses.
Keywords Atomic absorption
spectrometer Flame Uncertainty
of instrument

Also an example of estimation of uncertainty in AAS was

introduced in the Eurachem/CITAC Guide [5]. However,
in these works, most of the discussions concentrated on
the uncertainty of sample preparation procedures. Despite
the fact that the uncertainty of instruments is thought to be
important, it was treated as one factor and the uncertainty
sources of the instrument have not been analyzed further.
In some reports, the estimation of uncertainty for the instrument was limited to those concerning the calibration
of the instrument, while the instrument itself was considered as a black box.
We studied the uncertainty in flame AAS. In estimating the uncertainty of the atomic absorption spectrometer,
we considered the measurand of the atomic absorption
spectrometer is a value of related absorbance. Therefore,
in this study, we identified uncertainty sources affecting
measurement of the absorbance with the instrument, and
evaluated those sources.

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Fig. 2 Uncertainty sources for absorbance measurement in atomic

absorption spectrometry

Fig. 1 Schematic diagram of the atomic absorption spectrometer

Materials and methods

Hitachi Z-5010 and Z-2000 atomic absorption spectrometers were
used. The construction of the Z-5010 instrument is shown in Fig. 1.
Although the Z-2000 is a new model, the main constructions of the
two models are almost the same.
In Fig. 1, the light beam from the hollow cathode (HC) lamp is
passing through a flame above the burner head, mirrors, slits and a
grating, and finally reaches the photomultiplier tube (PMT) as a
detector. The signal from the PMT is amplified and converted into
absorbance units by electric circuit boards, and is transferred to a
control PC.
A Zeeman background correction system is used. It can be
considered as double-beam optics. However, in this study, do not
discuss the background correction by the Zeeman method. We
would like to discuss only the uncertainty concerned with singlebeam and double-beam measurements. We call the non-Zeemancorrected signal the measured signal obtained with single-beam
optics and the Zeeman-corrected signal the measured signal obtained with double-beam optics.
Cadmium was selected as a element showing a wide linear
range of the calibration curve, or the working curve, in AAS. Occasionally, molybdenum was used for comparison as an element
which shows opposite character to that of Cd. A reference solution
was prepared by diluting a commercially available reference solution for AAS.

Uncertainty sources
We assessed three main uncertainty sources for the
measurand of absorbance: drift of signal, noise, and
nonlinearity of the calibration curve. Each source is
considered to be influenced by several factors. The relationship is summarized in Fig. 2.

Fig. 3 Drift of baseline of Cd absorbance just after the lamp has

been turned on: a Single-beam measurement; b double-beam
measurement

Quantification of uncertainty sources

Drift of signal
Drift of lamp intensity
The light emission intensity of the HC lamp changes
shortly after it is turned on. The intensity of the lamp is
also changed by the temperature of the room and fluctuation of air surrounding the cathode of the lamp. The drift
of the Cd lamp intensity is shown in Fig. 3. The data were
obtained in a condition such that the cover of the lamp
housing was closed and the temperature of the air around
the lamp would not be disordered. The drift of the lamp
intensity directly affects the stability of the baseline in a
single-beam measurement. Double-beam optics can correct the drift of the light intensity and the baseline is
stable.
Figure 4 shows the drift of the Cd lamp intensity in
conditions such that the air surrounding the HC lamp was
disordered after a lamp warming-up time of 30 min. Note
that the ordinate is enlarged compared with that in Fig. 4.
The drift of the baseline in single-beam measurements is

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Fig. 5 Drift of Cd sensitivity just after turning on the lamp (0.5 mg/
l Cd)
Fig. 4 Drift of baseline of Cd absorbance after the lamp has
warmed up: A lamp housing closed, B lamp housing open, C: airblown. a Single-beam measurement; b double-beam measurement
Table 1 Uncertainty (absorbance units) of the drift of the lamp
intensity just after turning it on
Optics

020 min

2040 min

4060 min

Single beam
Double beam

0.07028
0.00042

0.00189
0.00018

0.00564
0.00011

not so different in conditions when the lamp housing of

the instrument is open or closed. However, when the HC
lamp was air-blown, the variation of the baseline became
large. In all cases, on the other hand, double-beam optics
was able to correct the drift of lamp intensity and the
baselines were very stable.
Table 1 shows the results of calculations of the uncertainty of the drift of the lamp intensity just after the
lamp has been turned on. The data correspond to those in
Fig. 3 and the uncertainties are expressed as standard
deviations of absorbance in each 20 min. In the singlebeam measurement, the uncertainty was very large in the
first 20 min, and became smaller in next 20 min. However, the uncertainty did not become much smaller in 40
60 min. The uncertainty did not remain constant. These
phenomena were typical in the single-beam measurement.
In the double-beam measurement, the uncertainty was
small in the first 20 min, and became smaller in next
20 min. Therefore, we should usually start measurements
about 20 min after turning on the lamp. The data obtained
with double-beam optics 20 min after turning on the lamp
were used later for calculation of the combined uncertainty.
Drift of sensitivity
The drift of Cd sensitivity is shown in Fig. 5. The absorbance values for 0.5 mg/l Cd were collected for 60 min
after the lamp had been turned on for the double-beam
measurement. The drift of the lamp intensity could be
corrected in the double-beam measurement as shown

Table 2 Uncertainty of the drift of the sensitivity for 0.5 mg/l Cd

(absorbance units) just after turning on the lamp
020 min

2040 min

4060 min

0.00382

0.00036

0.00016

Fig. 3. However, the sensitivity changed just after the

lamp was turned on. The decrease in the sensitivity is
caused by self-absorption of the resonance emission line
of the HC lamp. The result shows that 1020 min is required to reach a temperature equilibrium in the HC and
its surrounding area in the HC lamp.
Table 2 shows the uncertainty for the drift of Cd
sensitivity.
To obtain reliable data with smaller uncertainty, 20
40 min is required after turning on the lamp.
Noise
Flickering of flame, lamp energy and noise of the PMT
The baseline noise for Cd is shown in Fig. 6. The data
were collected for each 5 s of data measuring time with
different C2H2 flow rates, while the air flow rate was
fixed to 15 l/min. When the flame was turned off, the
noise became relatively small. This noise was considered
as the result of a variation of HC lamp energy and PMT
noise. When the flame was ignited, the noise became
greater and changed according to the increase in fuel flow
rate. These noises were considered as the result of the
flickering of the flame.
Table 3 shows the uncertainty for baseline noise.
Standard deviations of absorbance were calculated from
each 20 ms of data of computer output. When the flame
was turned off, the uncertainty changed little. When the
flame was ignited, the uncertainty increased especially in
a fuel-rich condition because of flickering of the flame.
The optimum fuel flow rate for Cd was 2.0 l/min.
Therefore, the uncertainty at that condition was used later
for calculation of the combined uncertainty. Also, this

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Fig. 6 Baseline noise for Cd:

a flame off; b fuel 2.0 l/min;
c fuel 2.6 l/min; d fuel 3.6 l/min

Table 3 Uncertainty (absorbance units) for baseline noise of Cd

Flame off

Fuel 2.0 l/min

Fuel 2.6 l/min

Fuel 3.6 l/min

0.00005

0.00007

0.00013

0.00028

Table 4 Uncertainty (absorbance units) for baseline noise of Mo

Flame off

Fuel 3.4 l/min

Fuel 3.7 l/min

Fuel 4.0 l/min

0.00003

0.00009

0.00011

0.00010

uncertainty includes uncertainties for lamp energy and

PMT noise.
Figure 7 and Table 4 are other examples for the
baseline noise of Mo. In contrast to Cd, a fuel-rich condition is preferable for Mo. Figure 7 shows that the noise
was small when the flame was turned off, and became
large when the flame was ignited. However, the noise was
not so different even when the fuel flow rate was changed.
Accordingly, the uncertainties were not so different even
when the fuel flow rate was varied as shown in Table 4.

Fig. 7 Baseline noise for Mo:

a flame off, b fuel 3.4 l/min,
c fuel 3.7 l/min, d fuel 4.0 l/min

Log transformation
The signal output from the PMT is transformed to absorbance in an electric circuit. The error in the transformation process may affect the uncertainty of absorbance.
In the log transformation process in the instruments we
used, light energy, which correspond to 100% transmittance, is adjusted to 3,000,000 counts of an analog-todigital converter output. Therefore, the electrical 1
count error is calculated as log(2,999,999/3,000,000)=
0.00000014 aborbance units. This means that the uncertainty for the log transformation process was negligible.
Nonlinearity of calibration curve
Line profile
In AAS, the line profile of the HC lamp and the absorption line profile are not exactly the same for some reasons
[6]. Doppler broadening is caused by thermal motion of
atoms. Because the temperatures of the lamp and the
flame are very different, the effects are different for

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Fig. 9 Theoretical calibration curve curvature caused by stray light.

Stray light a 0%, b 1%, c 2%, d 5%

Fig. 8 Relationship between the light emission profile and the

atomic absorption profile

emission and absorption line profiles. Pressure broadening is caused by collisions of atoms of different species or
of the same species. The pressure inside the HC lamp is
different from the pressure in the flame. Therefore, the
effects are different for emission and absorption line
profiles. Also, this effect acts to shift the profile to longer
wavelength. As a result, the relationship between the
emission profile and the absorption profile becomes like
that in Fig. 8 [7]. This relationship causes the calibration
curve bend. Moreover, self-absorption of the emission
line is caused by atomic absorption inside the HC lamp.
This phenomenon also acts to bend the calibration curve.
These are well-known phenomena in AAS.
However, the calibration curve bending caused by the
difference of the line profiles occurred at over about 0.3
absorbance units. Therefore, when the measurement is
performed within 0.3 absorbance units, the uncertainty of
the calibration curve nonlinearity caused by these phenomena will be negligible.
Stray light
Theoretical calibration curve curvatures caused by stray
light are shown in Fig. 9. The stray light has been discussed in for conventional spectrophotometry. When
there is stray light, the curvature becomes significant with
increasing stray light intensity. However, this effect is
prominent only in the region of absorbance larger than
1.0. Atomic absorption analysis is usually done in a region below 1.0 absorbance units, and it may be done
below 0.1 absorbance units. In addition, stray light for
recent spectrophotometer models is less than 0.05%.

Fig. 10 Burner optics in the flame of the atomic absorption spectrometer

These facts mean that extremely low stray light optics is

not required for AAS, and the uncertainty caused by the
stray light is negligible.
Flame condition and optics
The temperature in the center of the flame is highest and
the temperature in the outer region is lower. According to
the temperature distribution, the atomic density of the
analyte is not uniform in the flame. For example, Mg
shows a large area of high atomic density in the flame and
the shape of its contour lines of the density is gentle. The
contour lines are not so different between a fuel-rich
condition and a fuel-lean condition. Mo shows a small
area of high atomic density and the shape of the contour
lines of the density is steep. Also, the contour lines are
different between fuel-rich and fuel-lean conditions [8].
Figure 10 shows the light beam path from HC lamp in
the flame. The light beam is focused in the center of the
flame. As a result, the beam passes diagonally through
high and low atomic density areas in the flame. This

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Fig. 11 Calibration curves for Cd and Mo with different fuel flow rates. a Cd: a fuel 2.0 l/min; b fuel 3.6 l/min. b Mo: a fuel 3.4 l/min; b
fuel 3.7 l/min; c 4.0 l/min

phenomenon exists in the horizontal and vertical directions. This fact may cause nonlinearity of the calibration
curve. The contour line also changes when the flame
condition is changed; therefore, the uncertainty of the gas
flow rate will affect the uncertainty of absorbance.
Figure 11 shows calibration curves for Cd and Mo with
different fuel flow rates. In both cases, the calibration
curves bend in the high absorbance region. In the fuel-rich
condition, the sensitivity of Cd decreased and the calibration curve bending became prominent. The bending of
the Mo calibration curve was more prominent with a
small change of the fuel flow rate.
For the data shown in Fig. 11, the calculation of the
uncertainty at 0.5 mg/l Cd was performed as follows:
1. The absorbance of 0.5 mg/l Cd is 0.0782 at a fuel flow
rate of 2.0 l/min, and 0.0432 at 3.6 l/min. Then, the
absorbance change is 0.0219 for 1 l/min fuel difference.
2. The specification of the gas flow controller is 5%.
Then, an optimum flow rate of 2.0 l/min will be
controlled within 0.l l/min. Assuming a rectangular
distribution,
the uncertainty is estimated to be
p
0:1 3 0:0577l=min.
3. The corresponding uncertainty of absorbance will be
0.00126.
The calculation of uncertainty for Mo was done in the
same manner, resulting in 0.0222 absorbance units at
100 mg/l Mo.

Wavelength selection
Figure 12 shows the emission line spectrum of HC lamps
for six elements including typical complicated profiles of
Co, Fe and Ni. These line spectra include the instrumental
function (slit width); therefore, they were not substantially different from the line profile shown in Fig. 8. Most
of the line profiles for element not shown in Fig. 12 have
simple profiles. In early models in AAS history, the
wavelength had to be adjusted manually, and that could
seriously affect the uncertainty of the measurement.
However, in the latest instruments, the computer automatically searches the wavelength and sets the wavelength at the highest light intensity. Moreover, with the
development of high-performance HC lamps, the intensity
of the main resonance lines became higher than that of
neighboring lines. Therefore, the uncertainty caused by
wavelength selection is considered to be negligible.

Combined uncertainty
All the deviations caused by the previously mentioned
factors are considered to be incorporated in the measurand
(absorbance) by the linear combination. Therefore, the
combined uncertainty should be calculated from the
square root for the sum of square of each uncertainty. The
calculation is summarized in Table 5. The combined
uncertainty was estimated as 0.00133 at a Cd concentration of 0.5 mg/l (0.078 absorbance units). It was found

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Fig. 12 Emission line profiles of hollow cathode lamps: a Ca; b Cd; c Co; d Fe; e Mg; f Ni
Table 5 Estimation of combined uncertainty for Cd measurement

Uncertainty source

Uncertainty

Drift of lamp energy

Drift of sensitivity
Flickering of flame
(lamp energy and photomultiplier tube noise)
Log transformation
Line profile, stray light, wavelength selection
Flame condition/optics
Combined standard uncertainty=
(0.000182+0.000362+0.000072+0.001262)0.5

0.00018 absorbance units

0.00036 absorbance units
0.00007 absorbance units

that the contribution of the flame conditions and optics

was the largest. Therefore, the matrix effect of the sample
must be taken into consideration. For bending of the
calibration curve, a more reliable multicalibration curve

Negligible
Negligible
0.00126 absorbance units
0 00133 absorbance units
(0.5 mg/l Cd , 0.078 absorbance units)

method has also been proposed [9]. We will not express

an expanded uncertainty because our measurand was
absorbance in this report. We limited the discussion to the
uncertainty of the instrument.

References
1. Kurfrst U, Rehnert A, Muntau H (1996)
Spectrochim Acta B 51:229
2. Stringari G, Pancheri I, Mller F, Failla
O (1998) Accred Qual Assur 3:122
3. Bettencourt da Silva R, Cames M,
Seabra e Barros J (1998) Accred Qual
Assur 3:155

4. Synek V, Subrt P, Marecek J (2000)

Accred Qual Assur 5:58
5. Eurachem/CITAC Guide (2000) Quantifying uncertainty in analytical measurement. 2nd edn
6. Vandecasteele C, Block C (1993) Modern methods for trace element determination. John Wiley & Sons, New York

7. Yasuda K (1966) AnalChem 38:592

8. Rann C, Hambly A (1965) Anal Chem
37:879
9. Yasuda K, Imai K (1999) CITAC 99
Abstract p 56