ANALYTICAL SCIENCES FEBRUARY 2005, VOL.

21 143
2005 © The Japan Society for Analytical Chemistry

Sample Preparation for Evaluation of Detection Limits

in X-ray Fluorescence Spectrometry
M. K. TIWARI,† A. K. SINGH, and K. J. S. SAWHNEY

Synchrotron Utilization Division, Centre for Advanced Technology, Indore-452 013, India

The influence of analyte mass concentration on determination of detection limits in X-ray fluorescence spectrometry has
been investigated experimentally. Both the total reflection X-ray fluorescence (TXRF) and the conventional energy-
dispersive X-ray fluorescence techniques have been used to derive the dependence of analyte mass concentration on the
values of detection limits. Results obtained indicate that values of detection limits are optimum, or in other words, they
are closer to the true detection limit of the technique, when analyte concentrations are in the range of 10 times of the
detection limit.

(Received June 30, 2004; Accepted September 10, 2004)

described in detail by Pajek et al. They employed the process

Introduction
X-ray fluorescence spectrometry is one of the widely used
techniques for elemental analysis of materials. Using the
energy-dispersive X-ray fluorescence (EDXRF) analysis
technique, non-destructive multielement analysis at ppm levels
can be performed for elements ranging from sodium to uranium
in the periodic table.1,2 The technique finds several applications
to a variety of fields such as archeological, geological and
environmental.3–6 Total reflection X-ray fluorescence (TXRF)
technique is a variant of the conventional EDXRF technique,7–9
where the principle of total external reflection is used to restrict
the penetration of the primary beam into the specimen substrate.
This yields a substantial reduction of scattered background,
which improves the detection limit of the system to sub-
ppm/ppb levels.
The term “detection limit” (DL) is an important aspect of any
technique because it is an indication of the sensitivity of the
technique. As the name itself indicates, the detection limit is a
measure of the minimum quantity that can be detected by an
instrument or technique under the given experimental
conditions (sample geometry, sample matrix, analysis time,
etc.). A through description about detection limits can be
obtained from publications of Currie et al.10–14 The detection
values for an element are largely influenced by these
parameters. For example, in a multielement sample, the
presence of several peaks in the fluorescence spectrum can
modify the spectral background and hence change the DL value
of the element of interest. Such a limitation sometimes results
in “non-detects” i.e. some concentrations that cannot be
measured directly.15 The problem of non-detects often occurs
for biological and geological samples where the sample matrix
strongly influences the spectral background. The presence of
non-detects in XRF and TXRF analysis techniques has been
†
To whom correspondence should be addressed.
E-mail: mktiwari@cat.ernet.in
K. J. S. S. present address: Diamond Light Source Ltd., Rutherford
Appleton Laboratory, Chilton, Didcto-OX11 0QX, UK.
of “Censoring” for determining the detection sensitivities in
TXRF. The parameters that improve the detection limits of
various X-ray spectrometric techniques, such as PIXE,
WDXRF, synchrotron excited XRF and γ-ray spectroscopy,
have been discussed thoroughly by several workers.16–19
Furthermore, it has also shown that in X-ray tube excited XRF,
the detection limit can be improved by properly selecting the
primary filters.20 Moreover, a more elaborate theoretical
description for detection limit calculation in TXRF with various
modes of analysis, viz. grazing incidence (TXRF) and grazing
emission (GE-XRF) conditions, has been given by Sanchez.21
With developments/innovations in the fields of X-ray sources,
sample preparation, nuclear electronics, detection systems and
data processing procedures in the last decades, the XRF
techniques have also been refined to give better and better DLs.
Synchrotron radiation based X-ray fluorescence (SR-XRF) and
total reflection X-ray fluorescence (SR-TXRF) techniques are
the examples of such trends, where parts per trillion (ppt) level
or, in absolute mass, femto gram detection sensitivities have
been achieved.22–24 Normally, XRF techniques are comparative
in nature and type-standards are generally used to calibrate the
spectrometers. Similarly, synthetic standards are employed to
determine the DLs. In the literature, no guidelines are available
for the selection of analyte concentration to be used for detection
limit determination. Our experience, over the years, has been
that the DLs tend to depend on the analyte concentration in the
synthetic standard employed to determine the DLs.
We have undertaken a systematic study to resolve this issue;
our results give guidelines for what the analyte mass
concentrations should be for achieving the best possible DLs
under the given experimental conditions. This study indicates
that, to derive the optimum detection limit values, one should
locate the analyte mass concentration in the synthetic standards
in the range of 10 times the detection limit.
Theoretical Background
Mathematically, as per the IUPAC recommendation, the
144 ANALYTICAL SCIENCES FEBRUARY 2005, VOL. 21

Fig. 1 Variation of Zn detection limits with analyte concentration determined using EDXRF
technique for different acquisition times. Excitation source: Cd109. a, Solid square (s); b, solid circle
(s); c, solid triangle (g) show the measurements performed for acquisitions time 1000, 5000 and
10000 s, respectively whereas a solid line (m) shows the polynomial fit.

detection limit has been defined in terms of standard counting of analyte. From Eq. (1), it is clear that, for a specified counting
error of the background intensity.1 If σP and σB are the counting time, the minimum detectable concentration of analyte or
errors of the individual peak and the background, respectively, detection limit depends mainly on two parameters: the
then the standard counting error for a net count can be background intensity IB and the net area intensity per unit
expressed as, analyte mass (IA/CA).
—————
σ = √ σ P2 + σ B2
Experimental
In the limit of detection one can assume σP ≈ σB. Therefore
Both the EDXRF and TXRF techniques have been used to
——
σ ≈ √ 2σB perform X-ray fluorescence measurements. For EDXRF, an in-
house developed EDXRF spectrometer25 has been employed
For 95% confidence level, the counting error would be that comprises of a Si(Li) detector of energy resolution ∼155 eV
at 5.9 keV and an EG&G ORTEC 92X spectrum master that has
——
2σ ≈ 3ρB = 3 √ NB a built-in spectroscopy amplifier, a multi channel analyzer and a
HV bias supply. A 25 mCi Cd109 annular radioisotope is used as
By taking into account the counting time T and the slope m of an excitation source. Several synthetic standards, ranging in
fluorescence intensity vs. analyte concentration curve, one can three decades of analyte concentrations, were prepared. Zn and
write the expression for minimum detectable concentration for Cr elements were chosen as the analyte elements and their
an analyte element as synthetic standards were prepared in the form of pellets of mass
∼100 mg/cm2, by mixing their appropriate salts in cellulose
3 NB matrix. Moreover, to confirm the reproducibility of system
CDL = m
T geometry, we prepared synthetic standards of various elements
in various concentrations range and we analyzed them
or separately. The system geometry was found to reproduce
within ±0.5%. To perform the same studies using the TXRF
technique, researchers have used the indigenously developed
CDL = 3 IB (1)
 IA  TXRF spectrometer.26 Aqueous residues of U of different
 CA  concentrations were prepared from liquid solutions and polished
Si substrates were used as the sample carriers. Mo X-ray tube
where, IB = background intensity (NB/T); IA = net area intensity; (17.4 keV) was used as the excitation source.
CA = mass concentration of analyte; NB = background counts in
a given time T; m = IA/CA, i.e., slope of analyte counts-
concentration curve; CDL = minimum detectable concentration
ANALYTICAL SCIENCES FEBRUARY 2005, VOL. 21
Fig. 2 Variation of relative uncertainty for a signal at different
analyte concentrations (expressed in DL units) for two different risk
levels, 5% and 0.05%, calculated using Eq. (2). The variation of self-
absorption, A-factor, for Zn-Kα signal at different analyte
concentrations is also shown in this figure. The figure shows that for
analyte mass ranging in 10 – 30 times of DL value, both self-
absorption and relative uncertainties are optimum.
Results and Discussion
Figures 1a, b and c show the variation of detection limits of Zn
with various concentrations of Zn for analysis times of 1000 s,
5000 s and 10000 s, respectively. Since the DLs depend
inversely on the square root of analysis time, the DLs are better
for larger analysis times (Fig. 1). The figure shows that the
dependency of DL on analyte mass concentration generates
broad minima and that the DLs are minimum for analyte
concentrations ranging from 10 – 30 times of DL. Moreover,
one can see that the selection of an analyte concentration about
10 times that of DL is more appropriate for getting the
minimum DL value. For example, for 1000 s (5000 s) analysis
time, the minimum of DL for Zn is ∼59 (∼27) ppm which
corresponds to analyte mass of 600 (300) ppm. Similarly, for
analysis time of 10000 s, Zn DL is minimum (∼18 ppm) for the
analyte mass of 200 ppm. The appropriateness of analyte
concentration corresponding to 10 times that of DL can be
explained by the Currie hypothesis of “third detection limit”.10
Beside detection limit, which is a characteristic of a
measurement, Currie given a concept of third detection limit,
“determination limit”, LQ, above which the precision of a
measurement can be made good enough. The relationship
between determination limit LQ and detection limit CDL can be
approximated by16
CDL
LQ = ——
2kr
where, k defines the risk factor for a signal in terms of a fractile
of a Gaussian distribution and r is the required relative
uncertainty.
If LQ is set to n times of CDL i.e. LQ = nCDL then one can derive a
relation,
1 sample. By optimizing the width of the fluorescence peak
r = —— (2)
2nk
145
Fig. 3 Influence of analyte concentration on the background
intensity and slope m of analyte fluorescence intensity–concentration
calibration curve. Analysis time, 10000 s; analyte, Zn; slope m, (å);
background intensity, (©).
In the limit of detection where self-absorption effects27 for
analyte elements are significantly low, LQ simply represents the
smallest analyte mass that can be expected to generate a net
signal intensity with a relative error less than a selected limit r.
This can be understood more clearly from Fig. 2. Here, the
relative uncertainty for a signal at different levels of analyte
mass (expressed in DL units) has been plotted for two different
risk level 5% (k = 1.645) and 0.05% (k = 3.291) along with the
self absorption of Zn-Kα signal. For analyte masses ranging
around 10 – 30 times of DL, the relative uncertainties are
significantly low, e.g. comparable to acceptable limits of
statistical errors (∼0.5 – 2%) and at the same time self-
absorption effects do not affect severely the analyte signal
intensity. Beyond this range, at higher analyte concentrations,
though the relative uncertainties improve, the sample self-
absorption effects become dominant. Figure shows that
selecting analyte concentration in the range of 10 – 30 times of
DL would yield a fluorescence signal under acceptable
uncertainties. Self-absorption effects for analyte signal can be
ignored in this range. Though one can use any analyte mass
between 10 – 30 times of DL value for detection limit
measurement, it is a common practice to use an analyte mass
close to the DL value i.e. ∼10 times of DL.
The applicability of analyte mass corresponding to 10 times of
DL is shown in Fig. 3. Here, the net area intensity per unit
analyte concentration and the background fluorescence intensity
have been plotted as a function of Zn analyte concentration.
This figure shows that net area intensity of Zn-Kα per unit
analyte concentration, also referred to as the slope in Eq. (1),
show a peaking behavior for Zn analyte concentration of ∼200
ppm. On the other hand, the background intensity increases
monotonously, though this increase is very small. This is
because, at higher analyte concentrations, apart from sample
absorption effects, the tailing of the fluorescence peak increases,
leading to higher background below the fluorescence peak,
which in turn reduces the value of slope m. If one lowers the
analyte concentration, the background that appears due to
increase of tailing width of the fluorescence peak can be
reduced significantly. In this case it becomes almost constant
and equal to the value that one obtains in case of a blank
within 3σ limit and keeping background line well-known in the
area of the peak, the optimum value of m can be obtained for
146
Fig. 4 X-ray fluorescence spectra recorded for various
concentrations of Zn analyte. Analysis time, 10000 s; excitation
source, Cd109 radioisotope source. The figure shows that background
region/intensity under a fluorescence peak increases non-linearly
with increasing analyte concentrations.
analyte mass ∼10 times of DL. Figure 4 shows a more
elaborated picture of this non-linear behavior of the background
intensity, wherein fluorescence spectra for various analyte
concentrations have been plotted. The figure shows that, at
large analyte concentrations the background region/intensity
under a fluorescence peak is higher compared to that at lower
analyte concentrations, at the same time the net area intensity
behaves linearly at lower concentrations, which is not strictly
true at higher analyte concentrations because of sample
absorption effects. Since slope m as well as the background
intensity IB governs the detection limit value as defined in Eq.
(1), the detection limits improve for analyte concentrations
ranging in 100 – 500 ppm and a minimum is obtained at analyte
concentration of ∼200 ppm. Further reduction of analyte
concentration leads to poor statistics of analyte peak, resulting
in less than optimum values for DL. It is often true at very low
analyte concentrations that the number of counts that appears
under a fluorescence peak sometimes may not be sufficient to
support the Gaussian approximation i.e. the shape of the
fluorescence peak no longer remains Gaussian; rather, it shifts
to a Poisson distribution. This not only results in large errors in
net area estimation but it also sometimes becomes difficult to
define properly a window/ROI for a fluorescence peak in a
multichannel pulse height analyzer (MCA) spectra. Although
the detection limit values determined for two analytes Cr and Zn
are found to be minimum for analyte concentrations ranging
from 10 – 30 times of DLs, the dependency of DL on analyte
concentrations still shows that the selection of analyte
concentration corresponding to 10 times of DLs would be better
for getting optimum values of DL.
To further investigate the influence of analyte concentrations
on detection limit values, an independent study using TXRF
technique has also been performed. The detection limit values
in TXRF technique are generally determined by employing
aqueous residue generated from synthetic or standard solutions.
We used various concentrations of synthetic-standard solution
of uranium for determining the detection limit of uranium using
this technique. The dependency of detection limit on analyte
concentration studied using TXRF technique, Fig. 5, reconfirmed
that the best DL is obtained for analyte mass concentration
corresponding to ∼10 times of DL value. For instance, if the
true detection limit of the TXRF technique for uranium is 10
ANALYTICAL SCIENCES FEBRUARY 2005, VOL. 21
Fig. 5 Variation of U detection limits with analyte concentration
determined using TXRF technique. Excitation source: Mo X-ray
tube, 40 kV, 25 mA. Open circles (A) shows the measurements
performed for an acquisition time of 500 s whereas the solid line
(m) shows the polynomial fit. The inset shows the variation of net
area intensity for uranium Lα (slope m) and the background intensity
at different analyte concentrations.
ppb, then 100 ppb (0.1 ppm) of analyte concentration would be
appropriate for determining this detection limit. In TXRF, as
we have used aqueous samples for determination of DLs, apart
from the reasons discussed above that influences background
intensity, there is a certain probability of increased surface
roughness at large volumes of analyte. The inset of Fig. 5
shows the variation of net area intensity and background
intensity as a function of analyte (uranium) concentration. It is
found that net area intensity varies like it does in cases of
conventional XRF measurements, while the increase of
background is more rapid at large analyte concentrations. This
is mainly due to relatively large surface roughnesses induced at
higher analyte mass/volumes.
We have also taken into account the total errors contributed in
DL measurement as well as in preparation of analyte
concentration. Since the detection limit depends linearly on
analyte concentration, the net error in these two parameters has
been calculated partially. We have assumed that all
measurement errors; viz., errors in net area intensity, sample
preparation error, statistical error, or geometry reproducibility,
influence the DL value and have been determined using
synthetic standards of known concentrations. The total error
contributed to DL measurement has been found to vary from 2 –
8%. The error is higher at low analyte concentration and in this
case the statistical errors are dominant. On the other hand, to
estimate errors in preparation of analyte concentrations, we
prepared three sets of synthetic standards and analyzed each
standard for each concentration of analyte element. The net
error in analyte mass concentration has been determined
indirectly by measuring the fluorescence intensity. The total
error in analyte concentration has been found to vary from 2 –
10 %.
Conclusions
The systematic study presented here gives guidelines for the
most suitable analyte mass concentrations for achieving the best
possible DLs in XRF techniques. The results obtained indicate
that the optimum value of detection limit will correspond to
analyte mass ~10 times of the best detection limit. The same
ANALYTICAL SCIENCES FEBRUARY 2005, VOL. 21
inference can also be drawn from the TRXF analysis techniques
results, thereby indicating widespread possible applications of
this guideline especially in calibration of XRF spectrometers for
their DLs under given experimental conditions.
Acknowledgements
The authors thank R. V. Nandedkar for fruitful discussions and
suggestions.
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27. Sample absorption correction factor, A, in X-ray fluorescence
1 – exp –  µ(E0) µ(Ei) 
sinφ + sinϕ  M
can be evaluated from A = ,
 µ(E0) + µ(Ei)  M
 sinφ sinϕ 
where µ(E0) and µ(Ei) represent sample mass absorption
coefficients at incident energy E0 and characteristic
fluorescence line energy of the element of interest. φ and ϕ
are incident and exit angles; M represents the mass of the
pellet in g/cm2. To determine the self-absorption correction
for analyte signal, normalization of A factor to that in case
of blank sample is needed (A = 1 represents no absorption
and A = 1.10 means 10% absorption of analyte intensity).