Radiation Protection Dosimetry (2009), Vol. 133, No. 4, pp. 234–239 doi:10.

1093/rpd/ncp027
Advance Access publication 18 March 2009

URANIUM ISOTOPIC RATIO DETERMINATION IN URINE USING

FLOW-INJECTION ICP-MS: A TOOL FOR EMERGENCY
MONITORING
Maria Luiza D. P. Godoy, Ligia M. Q. C. Julião and José Marcus Godoy*
Instituto de Radioproteção e Dosimetria, Comissão Nacional de Energia Nuclear, Av. Salvador Allende s/n,
CEP 22780-160 Rio de Janeiro, RJ, Brazil

Received October 20 2008, revised January 28 2009, accepted February 18 2009

An inductively coupled plasma mass spectrometry-based method is presented aimed at total uranium concentration and
atomic ratio determination. The method includes flow-injection uranium separation based on TRU Eichromw extraction chro-
matographic cartridges. The method was tested with urine interlaboratorial exercise samples and certified reference materials
(NBL-CRM-U020A and NBL-CRM-U050) providing reliable results. The proposed methodology was also applied to urine
samples obtained after an incident at a nuclear facility. The obtained 235U/238U and 234U/238U atomic ratios in the urine of
an exposed worker were equal to those observed for an enriched uranium solution from the same origin.

INTRODUCTION Herein, the objective of the present work was the

Nuclear industry workers are routinely and,
potentially non-routinely, exposed to uranium-rich
aerosols. Therefore, an occupational monitoring pro-
gramme is a common practice and includes, among
others, in vitro measurements, such as urine analysis.
In the past, the total uranium content in a given
sample was determined by chemical methods, such
as fluorimetry, and uranium isotopes were deter-
mined by alpha-spectrometry. On the one hand, the
measuring equipment is relatively simple and low
cost, but on the other hand, both methods involve
sample digestion and chemical separation, which are
quite time consuming. This last characteristic is an
important handicap during emergency situations.
Therefore, a large number of alternative analytical
techniques, such as inductively coupled plasma mass
spectrometry (ICP-MS), are gaining popularity in
this field.
Since 238U ICP-MS determination has almost no
isobaric interference and the detection limit is in the
range of tenths of nanogram per litre, it can be
applied to urine samples for uranium determination
after simple dilution(1 – 5). However, for isotopic
analysis, matrix separation is advisable in order to
pre-concentrate the sample, lower the detection limit
and enable the determination of isotopes, such as
235
U and 234U. Several off-line methods for uranium
separation are available in the literature(6 – 8),
however, on-line methods(9 – 11) are particularly
attractive since they allow automatisation and yield
high precision, lower blanks and lower reagent
consumption.
*Corresponding author: jmgodoy@ird.gov.br
development of an ICP-MS-based analytical method
aimed at the fast determination of uranium concen-
tration and isotopic composition in urine samples.
Monitoring results taken after an accidental intake
of enriched uranium by workers of a nuclear facility
are also presented.
METHODOLOGY
238
U concentration was determined by ICP-MS, as
described by Godoy et al. (12), by applying a quanti-
tative method and using thallium as an internal
standard. The determination procedure can be sum-
marised as follows: starting from a Perkin-Elmer
multi-elemental standard solution, a 238U calibration
curve was built with the following total uranium
concentrations: 1, 5, 10, 15 and 20 mg l21, using
thallium as an internal standard. A 0.5 ml urine
sample aliquot was taken; thallium was added as the
internal standard, diluted to 10 ml with 2% nitric
acid (HNO3), and the 238U concentration was deter-
mined via ICP-MS. The detection limit typically
achieved was 4 ng l21 urine.
Isotopic ratio (IR) determinations were performed
using a Perkin-Elmer ELAN 6000 ICP-MS using
the same methodology previously developed for
environmental samples(13). However, some changes
were introduced: instead of UTEVA cartridges, TRU
cartridges were used, since uranium is more effec-
tively retained on TRU than on UTEVA columns,
which is desirable when working with samples with a
high salt content, as is the case with urine(9,10,14).
For the uranium separation, the system was coupled
to a Perkin-Elmer flow-injection system equipped
with an automatic sample changer. The original

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 URANIUM ISOTOPIC RATIO

Figure 1. Flow injection system with the autosampler, also showing the tubing connections according to the applied valve
position.

Figure 2. Obtained 238U and 235U elution curves after loading with 45 ml of 0.1 mg l21 natural uranium.

Table 1. Applied uranium on-line separation schema.

Programme step Read Time (s) Pump speed (rpm) Valve position

Pump 1 Pump 2

Pre-sample 60 60 1
1a 99 120 1
2 99 60 1
3 20 120 2
4 Yes 40 120 2
5 40 60 2
6 60 60 1
Post-run 30 60 2

a
This step is repeated three times.
235
 M. L. D. P. GODOY ET AL.

Table 2. Description of programme steps.

Programme step Description of step

Pre-sample Column rinse solution is pumped through the column to waste at 4.5 ml min21 to prepare the column
for loading. Water is pumped to the nebuliser at 3.5 ml min21 through the bypass.
1 Sample is pumped through the column to waste at 6 ml min21. Analytes are loaded onto the column.
This step can be repeated several times depending on the sample solution volume. Water is pumped to
the nebuliser at 4.5 ml min21.
2 Column rinse solution is pumped through the column to waste at 4.5 ml min21 to remove residual
sample. Water is pumped to the nebuliser at 3.5 ml min21 through the bypass.
3 Eluent is pumped through the column to the nebuliser at 7 ml min21. This step is used to delay the
start of the read cycle until the analytes approach the nebuliser. Rinse solution is pumped to waste at
9.0 ml min21 through the bypass.
4 This is the same as Step 3 except that the ICP-MS read cycle is initiated.
5 Eluent is pumped through the column to the nebuliser at 3.5 ml min21. Rinse solution is pumped to
waste at 4.5 ml min21 through the bypass. The purpose of this step is to rinse any remaining analyte.
6 Column rinse solution is pumped through the column to waste at 4.5 ml min21 to rinse residual eluent
and prepare the column for loading the next sample. Water is pumped to the nebuliser at 3.5 ml min21
through the bypass.
Post-run The autosampler returns to the rinse position for 30 s to rinse the sample inlet line with water at
3 ml min21. Water is pumped to the nebuliser at 2 ml min21.

scheme was modified in order to avoid the introduc- Table 3. Perkin-Elmer SCIEX ELAN 6000 ICP-MS
tion of ammonium oxalate into the ICP-MS during operation conditions for both measurement modes: IR and
the steps when the valve remained in position 1 concentration.
(Figure 1). The obtained 238U elution curve, starting
with 45 ml of solution containing 0.1 mg U(NBL- Instrument settings
CRM-U050) l21, is shown in Figure 2. It is possible RF Power 1050 W
to see that 40 s is sufficient time to elute all the Nebuliser Cross-flow (Ryton)
retained uranium in the cartridge. Spray chamber Scott (Ryton)
Sample cone Nickel 1.1 mm
The urine sample solutions were prepared by Skimmer cone Nickel 0.9 mm
mixing 5 ml of urine sample with 10 ml of sub-boiled Plasma gas flow rate 15 l min21
HNO3 and letting the mixture stand overnight. After Auxiliary gas flow rate 1.2 l min21
this period of time, the mixture was diluted to 45 ml Nebuliser gas flow rate 0.9 l min21
with Milli-Qw quality water. The sample solution Data acquisition settings
(3 mol HNO3) was pumped through the cartridge, Scan mode Peak hop transient
followed by a washing step with 3 mol HNO3 and Lens scanning Enabled
uranium was eluted using a 0.05 mol ammonium Detector mode Dual—pulse and analog
oxalate solution, as shown in Table 1. The loading Resolution 0.7 amm at 10% peak
maximum
step was repeated several times, depending on the Signal processing Spectral peaks integrated
volume of the sample solution, and each step com- Integration type Average
prised approximately 10 ml of sample solution. Atomic ratio measurements
Tables 2 and 3 show the description of the flow injec- Isotopes measured 234
U, 235U, 236U, 238U
tion analysis system programme steps and the Sweeps per readings 1
Perkin-Elmer ELAN 6000 ICP-MS operation con- Readings per replicate 300
ditions for both concentration and IR measurements, Number of replicates 1
respectively. Dwell time 30 ms
The urine samples represent a 24-h excretion. Total time of analysis 12 min
Concentration measurements
Samples were collected in 2-l glass bottles, acidified Isotopes measured 205
Tl, 238U
with 10 ml of sub-boiled HNO3 and stored at temp- Read delay 30 s
eratures below 48C until analysis. Sweeps per reading 20
For testing the precision and accuracy of the Readings per replicate 1
uranium atomic ratio determination, NBL-CRM- Number of replicates 3
U020A, NBL-CRM-U050 and a natural uranium Dwell time 30 ms
standard solution were utilised. Total time of analysis 40 s

236
 URANIUM ISOTOPIC RATIO
The associated uncertainty for one single determi-
nation of the IR was calculated as follows:
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
1 1
Uncertainty ¼ IR  þ ; ð1Þ
C1 C2
where Ci is the total counts obtained for the
isotope i.
RESULTS
In order to test the proposed procedure, aliquots
from the NBL-CRM-U020A, NBL-CRM-U050 or
natural uranium standard solution were taken and
added to 45 ml of 3 mol HNO3, providing a concen-
tration level equivalent to 0.1 mgU l21, from which
the 236U/238U, 235U/238U and 234U/238U atomic
ratios were determined. Based on the results shown
in Table 4, it is possible to verify that high precision
and accuracy are achieved with the proposed
methodology.
The achieved accuracy and precision when dealing
with real urine samples were determined using the
2007 PROCORAD(15) intercomparison samples, as
shown in Tables 5 and 6. For the uranium content,
it is possible to verify that a precision level of 1–2%
was achieved for both concentration levels; accuracy
values as low as 2% were obtained at the 40 mg l21
level and around 10% even at the 2 mg l21 level.
However, it should be noted that there are differ-
ences between the reference values provided by
the intercomparison exercise organiser and the
mean values obtained by consensus among all par-
ticipants, 40.02 and 1.928 mg l21, for samples A and
B, respectively. When applying the consensus values
instead of the reference ones, the observed relative
error is 3–4% for both samples. As expected, due to

Table 4. Obtained precision and accuracy of the atomic ratio determination of certified solutions with a uranium
concentration of 0.1 mg L21 after chemical purification according the developed schema.

Atomic ratio Certified Measured Precision (%) Accuracy (%)

234
CRM U020-A U/238U 0.00017683 0.0001790 + 0.0000039 2.2 1.2
235
U/238U 0.02068718 0.020633 + 0.000042 0.20 20.3
236
U/238U 0.00012037 0.0001220 + 0.0000032 2.6 1.4
234
CRM U050 U/238U 0.0002939 0.000287 + 0.000016 5.5 22.3
235
U/238U 0.0527841 0.05073 + 0.00021 0.42 23.9
236
U/238U 0.0005057 0.000409 + 0.000019 4.6 219
234
Natural uranium U/238U 0.0000554 0.0000550 + 0.0000021 3.8 20.72
235
U/238U 0.0072526 0.007133 + 0.000024 0.33 21.6

Plus or minus denotes uncertainty of one determination.

Table 5. Uranium content during the PROCORAD 2007 interlaboratorial exercise.

Sample Uranium (mg l21) Precision (%) Accuracy (%)

a
Reference value Measured

A 38 38.06 + 0.58 1.5 0.16

B 2.02 1.866 + 0.022 1.2 27.6

a
Data represent the mean + 1 standard deviation of three analyses.

Table 6. Uranium atomic ratios during the PROCORAD 2007 interlaboratorial exercise in urine samples.

Sample Atomic ratio Reference value (natural uranium) Measureda Precision (%) Accuracy (%)

234
A U/238U 0.0000554 0.0000537 + 0.0000058 11 23.1
235
U/238U 0.0072526 0.007183 + 0.000072 1.0 2.3
234
B U/238U 0.0000554 0.0000567 + 0.000015 27 21.0
235
U/238U 0.0072526 0.007097 + 0.000048 0.67 22.1

a
Data represent the mean + 1 standard deviation of three analyses.

237
 M. L. D. P. GODOY ET AL.
Table 7. Observed uranium concentration and atomic ratios in the urine of two workers after an incident with an enriched
uranium solution, and the atomic ratio of this solution.
Sample U (mg L21)a
Worker no. 1 0.1180 + 0.0007
Worker no. 2 1.1895 + 0.0059
Feed solution
a
Mean + 1 standard deviation of three analyses.
b
Result + uncertainty of one determination.
the higher abundance of the 235U isotope in relation
to the 234U isotope, higher precision is achieved for
the 235U/238U than for the 234U/238U atomic ratio
(Table 6). Specifically, for the 235U/238U atomic
ratio determination, 1% precision is obtained. The
observed accuracy is similar for the 235U/238U and
234
U/238U atomic ratios in both samples with rela-
tive errors of 3% or below.
This procedure was tested under real circum-
stances. After an incident at a nuclear facility in
which two workers were exposed to an enriched
uranium feed solution, 24-h urine samples were col-
lected and sent to the laboratory for analysis. Table 7
shows the observed results in terms of total uranium
content and the 235U/238U and 234U/238U atomic
ratios in the urine samples. The 235U/238U and
234
U/238U atomic ratios in the enriched uranium sol-
ution are also shown. Taking into account a uranium
background level in urine of about 0.006 mg l21 for
individuals living in Southeastern Brazil proposed by
Santos(16), it is possible to verify that both workers
presented an increased uranium content in their
urine. The 235U/238U and 234U/238U atomic ratios for
both workers are quite different from the normal
ratios, 0.0072526 and 0.0000554, respectively. The
most contaminated worker presented 235U/238U and
234
U/238U atomic ratios equal to those observed for
the enriched uranium solution. Eighteen days later, the
second worker presented a 0.019 mg l21 uranium con-
centration in the urine, which allowed the estimation
of a urinary excretion half-life of 2.8 d. This half-life is
shorter than expected for acute ingestion of a soluble
compound or the inhalation of a class F material.(17)
This incident shows that the proposed method-
ology can be applied for personal monitoring pur-
poses after an emergency situation, providing fast
and reliable results to the authorities.
The separation, measurement and preparation for
the next sample takes 10.8 min, which gives an
analysis rate of 5.7 samples per hour. One cartridge
can be used up to 200– 250 times.(18) Normally,
urine is mixed with nitric acid and allowed to stand
overnight to digest; however, if necessary, microwave
digestion can be applied, for example, a programme
238
Atomic ratiob
234
U/238U 235
U/238U
0.000134 + 0.000024 0.02898 + 0.00036
0.000333 + 0.000011 0.03904 + 0.00013
0.000344 + 0.000007 0.03983 + 0.00036
with 10 min at 600 W and 1808C produces a clear
and colourless solution.
CONCLUSION
The proposed methodology for determination of the
uranium concentration and atomic ratio in urine
provides fast and reliable results, as was demon-
strated after a real-world contamination situation.
Based on this case, a urinary excretion half-life of
2.8 d was estimated.
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