9 1999by Humana Press Inc.

All rights of any nature, whatsoever, reserved.

0163-4984/99/71-72--0423 $12.00

Limits of Detection of a Total

Reflection X-Ray Fluorescence
System with Double Reflection
Module
V. F. NASCIMENTO FILHO,*'1'2 V. H. POBLETE,3
P. S. PARREIRA,1'4 E. MATSUMOTO,5 S. M. SIMABUCO,5
E. P. E S P I N O Z A , 1 AND A. A. NAVARRO1
lLaboratOrio de Instrumenta~o Nuclear/CENA/USP, av,
Centen~rio, 303, CP 96, CEP13400-970 Piracicaba, S~o Paulo,
Brazil; 2Departamento de Fisica e Meteorologia/ESALQ/USP,
av, P~dua Dias, 11, CP 9, CEP 13418-900, Piracicaba, S~o Paulo,
Brazil; 3Laboratorio de Fluorescencia de Rayos X/CCHEN,
Amunatequi, 95, Cassila 188-D, Santiago, Chile; 4Departamento
de Fisica/CCE/UEL, Campus Universit~rio, CP 6001, CEP 86051-
970, Londrina, Paran& Brazil; and 5Departamento de Recursos
Hidricos/FEC/UNICAMP, Campus Universitario, CP 6021,
CEP 13081-970, Campinas, S~o Paulo, Brazil

ABSTRACT
An X-ray tube with a Mo target and Zr filter, operated at 45
kV/20 mA, was used to excite samples (5 ~tL deposited on a quartz
support) and the total reflection angle condition was obtained with a
double reflector module built with two 10-cm-long 7-mm-thick quartz
crystals placed 50 ~m apart. A high-resolution spectrometer based on
a Si(Li) detector coupled to a multichannel analyzer was used for
X-ray detection and the spectra were interpreted with the AXIL
software.
The system was calibrated with standard chemical solutions con-
taining Cr, Fe, Cu, Zn, and Pb, and Y was used as an internal standard
to correct eventual geometric errors and high-voltage instabilities of
the X-ray generator. The limits of detection were 19, 9, 5, and 4 n g / m L
for Cr, Fe, Cu, and Zn, respectively, analyzed through characteristic Ks
X-rays, and 7 n g / m L for Pb, through L~ X-rays, considering 50 ~tL

*Author to whom all correspondence and reprint requests should be addressed.

Biological Trace Element Research 423 Vols. 71-72, 1999

424 Nascimento Filho et al.

samples deposited and dried on a quartz support, to be excited/

detected for 1000 s.
Index Entries: Total reflection x-ray fluorescence; TXRF; XRF;
ED-XRF; instrumental analysis.

INTRODUCTION AND OBJECTIVES

The total reflection X-ray fluorescence (TXRF) technique is a variance
of the conventional energy-dispersion X-ray fluorescence (ED-XRF) tech-
nique; it was introduced in 1971 by Yoneda and Horiuchi (1) and devel-
oped by Aiginger and Wobrauschek (2,3). It allows an instrumental,
simultaneous, and multielementary chemical analysis measuring the
intensities of characteristic X-rays emitted by the constituent elements of
the sample. This technique is based on the incidence of an X-ray beam at
a sufficiently small angle (called critical angle of total reflection) on the
flat surface of a support or carrier (e.g., quartz) on which the sample to
be analyzed is deposited. In this condition, the incident beam spreads
parallel to the surface of the support and is totally used in the excitement
of the elements of the sample, producing characteristic X-rays with no
Rayleigh or Compton effect (4) scattering.
Consequently, a spectrum of low intensities of the coherent and inco-
herent scattering peaks coming from the excitation source is obtained. In
addition, there is a reduction of the continuum area under the characteris-
tic peaks, elevating the peak-background ratio considerably, thus leading
to a consequent decrease in the limits of detection. Another characteris-
tic of this technique, when compared to the conventional technique, is
the use of small volumes of liquid samples (in microliters) or small
masses (in micrograms) of solid samples after chemical digestion, being,
therefore, adequate for the analysis of trace elements mainly in samples
for environmental monitoring, of biological fluids and for quality control
of high-purity products.
Because of the way it is prepared, the sample to be deposited on the
support has a very reduced thickness and the absorption and enhance-
ment effects can be negleted. In this case, the quantification of the chem-
ical elements can be made through the employment of Eq. (1), considering
the sample as a thin film:

I i = siC i (1)

where Ii represents the intensity (cps) of the K or L characteristic X-ray of

the interest element i, Ci represents its concentration (~tg/mL), and si is the
sensitivity of the system (cps/~tg/mL for this element.
The main objective of this work was the installation of a TXRF sys-
tem, which was nonexistent in Brazil, to determine the relative elemen-
tary sensitivities and the limits of detection for some elements of interest
in liquid samples.

Biological Trace Element Research Vols. 71-72, 1999

Total Reflection X-ray Fluorescence System 425

Fig. 1. Total reflection X-ray fluorescence system: visualization of the

tower with the X-ray tube (XRT), high-voltage generator (HVG), X-ray detector
(XRD), and the total reflection module (TRM).

MATERIALS AND METHODS

Nuclear Instrumentation
The detection of the X-rays was made using a Si(Li) semiconductor
detector coupled to a preamplifier, amplifier, and multichalmel pulse
analyzer card inserted in a microcomputer. The detector, manufactured
by EG & G Ortec, model SL 30165, is 3-mm-thick and has an active area
of 30 m m 2, 7.5-gm-thick a Be window, and a distance of 5 m m between
the Be window and the crystal surface, presenting a resolution of 165 eV
to 5.9 keV (Fig. 1).

Total Reflection Module

The module is basically constituted of a lead collimator with a 0.1-mm
aperture mounted by the Zr filter, a double crystal reflector module (com-
posed of two quartz sheets 7-mm-thick and 100-mm-long, placed in paral-
lel, 50 gm apart) and a sample holder (Fig. 1) at the exit of the reflector
system. This reflector system has three micrometric screws for the adjust-
ment of the incidence angle (total reflection) of the radiation on the sample
support. More details about the module can be found in ref. 5.

Biological Trace Element Research Vols. 71-72, 1999

426 Nascimento Filho et al.

Excitation System
The excitation system is composed of a fine-focus X-ray tube, with a Mo
target and Zr filter, coupled to a high-voltage generator operated at 45 kV
and 20 mA. The X-ray tower has four radiation exits (two point types and
two line types), at an angle distance of 90~ from each other, which can be
used simultaneousl~ given that one line type is used in the TXRF (Fig. 1).

Calibration of the Total Reflection Module

The total reflection module is calibrated to obtain the best geometric
configuration, so that the excitation radiation shines on the sample sup-
port in the condition of total reflection. The incidence angle for total
reflection is called the critical angle (I)crit (min) and can be calculated by
eq. (2), where E represents the energy of the incident radiation (in keV)
and p, Z, and A are the density (g/cm3), the number of electrons in an
atom or molecule, and the atomgram or mol (g/mol) of the sample sup-
port, respectively (6,7):

E ]PA
dilcrit = __99
(2)
For the system in use, with a Mo target X-ray tube and quartz (SiO2)
as the sample support, the parameters are p = 2.5 g/cm 3, Z = 30 elec-
trons/molecule, A = 60.0863 g/mol, and an energy of Mo Kc~ X-ray inci-
dent radiation E = 17.44 keV, yielding a critical angle of 6.4 min. This
angle is adjusted using the micrometer screws of the module reflector.

Stardard Solutions
To perform the calibration of the system, four standard solutions
containing Ca, Cr, Fe, Cu, Zn, Ba, and Pb were prepared at different and
well-known concentrations (ranging from 2 to 30 gg/mL, approximately).
Yttrium was added, always to a final concentration of 8.33 gg/mL, as an
internal standard to eliminate the errors caused by excitation/detection
geometry. The thin film obtained from the sample formed on the quartz
support does not have a regular geometry; therefore, the intensity of the
characteristic X-ray depends on the position of the thin film on the sup-
port. This geometric effect of the sample (8,9) can be corrected by calcu-
lating the relative intensity of each element in relation to this internal
standard [(Eq. (3)].

Preparation of Samples
for Excitation~Detection
An aliquot of 5 gL of each one of the standard solutions was pipeted
on a quartz support and dried under vacuum, forming a thin film (two
replicates for each standard solution). The samples were excited for 500 s
and the X-ray spectra obtained were fitted by the AXIL software. The net

Biological Trace Element Research Vols. 71-72, 1999

Total Reflection X-ray Fluorescence S y s t e m 427

1000

9. - . 80O
o
o

" 600
8

Z 400
uJ
m

~, 20o
)<

0
2 4 6 8 10 12 14 16 18 20
ENERGY (keV)

Fig. 2. Characteristic X-ray spectrum of the elements of one standard

sample.

areas of the characteristic X-ray peaks of the elements that compose the
samples were evaluated.

RESULTS AND DISCUSSION

Relative Intensities
As an illustration, Fig. 2 shows the X-ray spectrum of one standard
sample, where the concentration of the all elements is 8.33 ~tg/mL. With
the intensities of the characteristic X-rays obtained using standard sam-
ples, the relative intensities Ri were calculated using Eq. (3). It has to be
pointed out that the relative intensities Ri is not only the ratio between
the intensities of the measured element Ii and that of the internal stan-
dard Iy, because the concentration of the internal standard Cy (8,9) has to
be taken into account. In other words, using Eq. (1), the following equa-
tion can be written:

Ii _ Si Ci
Iy Sy Cy
or
I-Li C y = s--LC i
Ir Sy

Biological Trace Element Research Vols. 71-72, 1999

428 Nascimento Filho et al.

10 I I I I I I

R= 0,2EBOZn I]
~8 Zn r2 = 09993
v

[]

Cu R = Q2256Q
Z
~6 r2= 0.981
z
m

uJ R= 0.1422Fe
Fe r2 = Q9923
LU
OE
R=0.074t.Cr i
r2 = 99981

I r I I
5 1~ is 9 2s 30 35
CONCENTRATION (~g/mL)

Fig. 3. Linear regressions for the estimation of the relative elementary

sensitivities.

I I "" I I I I

~ 18 ~k~ LDO=054e6.Z2-31.860Z+4686
v
Z
O14
I-

I-

14.
O8
I-
$8-
,--I

4-
2~
Cr IVh Fe Co Ni Cu Zn
0 ! I I I I I I
23 24 25 ;6 27 28 29 30
ATOMIC NUMBER(Z)
Fig. 4. Limits of detection (ng/mL) for some elements in liquid samples
using 50 ~tL aliquots deposited on quartz supports and analyzed for 1000 s.

Biological Trace Element Research Vols. 71-72, 1999

Total Reflection X-ray Fluorescence System 429

I-L Cy
R i = [y

and because
Si si (3)
Sy

the result is
R i = SiC i (4)

where Ii and Ci are the intensity and concentration of element i in the

sample, respectively, Iy and Cy are the intensity and concentration of the
internal standard Y in the sample, respectively, si and Sy are the sensitiv-
ities of the system for the element i and for the internal standard Y,
respectively, Ri is the relative intensity for element i, and Si is the relative
sensitivity (adimensional) of the system for the element i.

Relative Elementary Sensitivities

Using the concentrations Ci of the elements and the relative intensi-
ties Ri, linear regressions shown in Fig. 3 were obtained for each element,
where the angular coefficients are the relative elementary sensitivities.

Limits of Detection
The limits of detection LODi (10,11) were calculated using Eq. (5) for
a analysis time of 1000 s and the results obtained are shown in Fig. 4. For
this, the background intensities Ii(BG) (i.e., the areas under the peaks of
the elements of interest), were obtained through the fitting of the spectra
by the AXIL software, the intensity Ir for the element Y was taken as one
obtained with the concentration of 8.33 ~tg/mL:

LOD i = 3~/ Ii(BG) Cy

t IySi (5)
It must be observed that aliquots of 5 ~tL were used for the cal-
culation of the limits of detection. These values could even be smaller.
Thus, if aliquots of 50 ~tL of aqueous samples are used, LODi would be
19, 9, 5, and 4 n g / m L for elements Cr, Fe, Cu, and Zn, respectively,
when analyzed through the characteristic KR X-rays, and 7 n g / m L for
Pb, analyzed through the characteristic L~ X-rays. As for solid samples,
such as soil, sediments, biological tissues, and so forth, when 10 mg of
the sample are previously digested chemically, completed to a volume
of 1 mL, and later using 50 ~tL in the analysis, the LODi would be 1.9,
0.9, 0.5, 0.4, and 0.7 ~ g / m L for the same elements. Another field for
the application of TXRF include studies of environment air contami-
nation; in this case, for the digestion of the filters, after collection of
a volume of air of 10 m 3, with a volume completion to 1 mL and

Biological Trace Element Research Vols. 71-72, 1999

430 Nascimento Filho et al.

use of 50 gL in the analysis, the LODi would be 1.9, 0.9, 0.5, 0.4, and
0.7 n g / m 3, respectively.

ACKNOWLEDGMENTS
The authors thank Funda~io de Amparo ~ Pesquisa do Estado de
Silo Paulo, Silo Paulo State Foundation for Support to Research (Procs.
95/0018-7 and 96/5818-4) for the financial support.

REFERENCES
1. Y. Yoneda and T. Horiuchi, Optical flats for use in X-ray spectrochemical micro-
analysis, Rev. Sci. Instrum. 42, 1069 (1971).
2. H. Aiginger and P. Wobrauschek, A method for quantitative X-ray fluorescence
analysis in the nanogram region, Nucl. Instrum. Methods 114, 157-158 (1974).
3. P. Wobrauschek and H. Aiginger, Total reflection fluorescence spectrometric determi-
nation of elements in ng amounts, Anal. Chem. 47, 852-855 (1975).
4. C. Koopmann and A. Prange, Multielement determination in sediments from the
German Wadden Sea--investigations on sample preparation techniques, Spectrochim.
Acta 46B, 1395-1402 (1991).
5. V. H. Poblete, Ph.D. Thesis, Facultad de Ciencias F~sicas y Matematicas, Universidad
de Chile, Santiago, Chile (1996), Chap. III, p. 48.
6. H. Aiginger, Historical development and principles of total reflection X-ray fluores-
cence analysis (TXRF), Spectrochim. Acta 46B, 1313-1321 (1991).
7. A. Prange and H. Schwenke, Trace element analysis using total-reflection X-ray flu-
orescence spectrometry, Adv. X-Ray Anal. 35, 899-923 (1992).
8. W. Ladisich, R. Rieder, and P. Wobrauschek, Quantitative total reflection X-ray fluo-
rescence analysis with monoenergetic excitation, X-ray Spectrom. 23, 173-177 (1994).
9. R. Klockemk/imper and A. von Bohlen, Elemental analysis of environmental samples
by total reflection fluorescence: a review, X-ray Spectrom. 25, 156-162 (1996).
10. L. A. Curie, Limits for quantitative detection and quantitative determination, Anal.
Chem. 40, 586-593 (1968).
11. W. Ladisich, R. Rieder, P. Wobrauschek, and H. Aiginger, Total reflection X-ray fluo-
rescence analysis with monoenergetic excitation using rotating anode X-ray tubes,
Nucl. Instrum. Methods Phys. Res. 330A, 501-506 (1993).

Biological Trace Element Research Vols. 71-72, 1999