Food Chemistry 132 (2012) 1555–1561

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Analytical Methods

Estimation of uncertainties of the method to determine the concentrations of Cd,

Cu, Fe, Pb, Sn and Zn in tomato paste samples analysed by high resolution ICP-MS
Serpil Yenisoy-Karakasß ⇑
TUBITAK-UME, National Metrology Institute, P.O. Box 54, 41470 Gebze-Kocaeli, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Cadmium, copper, iron, lead, tin and zinc were determined in tomato paste samples by using high reso-
Received 24 January 2011 lution inductively coupled plasma mass spectrometry. This study was a part of Proficiency Testing
Received in revised form 15 April 2011 scheme conducted by National Metrology Institute (UME) of Turkey. The method was validated. The per-
Accepted 9 November 2011
centage relative errors obtained for each element ranged between 1.4% and 9.0% for NIST SRM 1573a,
Available online 17 November 2011
tomato leaves. Measurement uncertainties of all elements were calculated by applying bottom-up
approach. The calculated percentage relative uncertainties for the elements were between 8.2% and
Keywords:
16.4%. The major contributions to the uncertainty budget came from the calibration curves, repeatability
Elements
Tomato paste
and recovery. Tomato paste samples gathered from the supermarkets in Turkey were analysed by using
High resolution inductively coupled plasma the validated method. Their levels were compared both with the literature values and European Commis-
mass spectrometry sion Regulations.
Method validation Ó 2011 Elsevier Ltd. All rights reserved.
Recovery
Uncertainty

1. Introduction
A variety of chemicals may enter the food chain by means of
intentional and unintentional addition. These chemicals include
many substances like food additives, pesticide residues, environ-
mental contaminants, mycotoxins, flavouring substances and
micronutrients. Packaging materials are also a source of chemicals
in food products and beverages. It is recognised that chemicals from
packaging and other food-contact materials can migrate into the
food itself and can be ingested by consumer (Poças & Hogg, 2007).
Metals like iron, copper and zinc are essential metals since they play
an important role in biological systems, whereas lead, cadmium and
tin are non-essential metals as they are toxic even in trace amounts
(Blunden & Wallace, 2003; Somers, 1974). Toxic elements can be
very harmful, even at low concentration, when ingested over a long
time period. The essential metal can also produce toxic effects when
the metal is taken excessively.
Canned tomato pastes have a popular usage in Turkey. How-
ever, data on trace metal levels in canned tomato paste produced
in Turkey and also in the world are very scarce if there are any.
There are some research conducted on determination of metals
on food (mushroom, corn, pea, mixed vegetables, tomato, stuffed
grape leaves, pickle, bean and delicatessen) (Tüzen & Soylak,
⇑ Present address: University of Abant Izzet Baysal, Faculty of Sciences, Depart-
ment of Chemistry, 14280 Bolu, Turkey.
E-mail address: yenisoykarakas_s@ibu.edu.tr
2007a) and canned fish marketed in Turkey (Tüzen & Soylak,
2007b).
Due to the importance of trace elements on human metabolism,
their analysis is an important part of public health studies. Because
of its high sensitivity, wide dynamic range and relatively low pos-
sibility of interferences as well as its multi elemental characteris-
tics, inductively coupled plasma mass spectrometry (ICP-MS) has
been chosen as the analytical technique in food matrices (Cubadda
& Raggi, 2005; Nardi et al., 2009). Although there are many advan-
tages of ICP-MS, it suffers from both spectroscopic and non-
spectroscopic interferences. However, mass spectral overlapping
with polyatomic ions, has been recently overcome by the use of
high resolution inductively coupled plasma mass spectrometer
(HR-ICP-MS). High resolution ICP-MS relies on a magnetic sector
to separate ions with greater discrimination according to their
mass/charge ratio. The introduction of high resolution (magnetic
sector) ICP-MS instruments has opened new perspectives in the
field of isotopic measurements. Typically, the repeatability on
isotope abundance ratio measurements that can be achieved in a
reproducible manner is 0.1–0.35% for quadrupole ICP-MS and
down to 0.05% for high resolution ICP-MS (Quétel et al., 2000).
It is unavoidable to calculate the measurement uncertainties
when the trace and ultra trace levels of elements are determined.
For this reason, it is necessary to identify all the possible sources
of uncertainties related to the analytical procedure. The Eurachem/
Citac (2000) guide provides examples on identifying uncertainty
sources and combining the relevant uncertainty contributions. The

0308-8146/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.11.047
1556 S. Yenisoy-Karakasß / Food Chemistry 132 (2012) 1555–1561
ISO/TS 21748 guide (2003) suggests using experimental data from
precision and trueness or recovery studies in the calculation of
uncertainty. The application to real samples is the next step after
the validation giving the evidence that the developed method can
be used in the laboratory in the linear range of the calibration curve
(Forte & Bocca, 2007).
There are some studies giving the estimation of uncertainties
(Forte & Bocca, 2007; Mestek, Komínková, Koplík, Borková, &
Suchánek, 2002; Trešl & Quétel, 2005; Vassileva & Quétel, 2004)
of metals using ICP-MS instrument in several matrices like food,
water, offal and using ICP-OES instrument in complex matrices like
alloys, geological materials (Rabb & Olesik, 2008). However all of
them just gave the resulting uncertainty values without giving
the details of calculations that is to say without the uncertainty
budget. One study conducted by Barwick, Ellison, and Fairman
(1999) is giving the details of uncertainty calculation of the con-
centrations of 60Ni in water samples. The calculations were given
in fully worked equations and very clear to follow.
The approach in uncertainty calculations differs significantly
since the methods and the matrices are very different from each
other even the people use the same technique. We have developed
a strategy for reconciling the existing experimental and non-
experimental data. This strategy results in a structured list of the
possible sources in uncertainty budget associated with method.
Although six metals are considered here, similar calculations can
be undertaken for other routinely analysed elements for which
QC and/or method validation data are available.
This study was started for method validation and uncertainty
calculation of PT (Proficiency Testing) test ‘‘metals in tomato
paste’’ conducted by TUBITAK-UME (The Scientific and Research
Council of Turkey-National Metrology Institute). However, it was
extended to sample the canned tomato pastes and to analyse them
with the validated method. The performance of HR-ICP-MS tech-
nique has been evaluated in terms of uncertainties of the measure-
ments in the determination of six elements in tomato paste
samples. One important contribution of this study is that as far
as we know there is no other study in the literature related with
the determination of Sn in food matrix with the technique of HR-
ICP-MS or quadrupole ICP-MS.
2. Experimental
2.1. Samples
The canned tomato pastes with different trade marks (Turkish)
were purchased from the supermarkets during the year of 2007 in
the cities of Istanbul and Kocaeli. Fourteen cans gathered from the
supermarkets were used for the analyses. NIST SRM 1573a (tomato
leaves) certified reference material (CRM) issued from National
Institute of Standard and Technology (NIST, Gaithersberg, MD,
USA) was used for the trueness.
2.2. Instrumentation and operating parameters
The analyses were performed by using Thermo Finnigan Ele-
ment 2 (Bremen, Germany) high resolution ICP-MS instrument.
The operating parameters are summarised in Table 1. The isotopes
and mass resolution of elements used in the analyses were as fol-
lows: 111Cd(LR), 114Cd(LR), 63Cu (LR, MR), 65Cu (LR, MR), 56Fe (MR,
HR), 57Fe (MR, HR), 206Pb (LR, MR), 208Pb (LR, MR), 118Sn (LR, HR),
66
Zn (LR, MR) and 68Zn (LR, MR). The quantification of Fe was car-
ried out in both medium resolution and high resolution modes. In
fact, the most abundant isotope at mass 56 is strongly affected by
the interfering biatomic species 40Ar16O. In the case of quadrupole
ICP-MS instrument, the very high amount of Ar and O present in
the plasma makes the use of the mathematical equation based
on the 18O signal inapplicable. This problem can be solved by
HR-ICP-MS at a mass resolution of medium and high (D’llio, Ales-
sandrelli, Cresti, Forte & Caroli, 2002).
2.3. Reagents and standards
All reagents used in the analysis are analytical or higher grade.
Suprapure nitric acid (65%, w/w) and analytical grade hydrogen
peroxide (30%, w/w) were purchased from Merck (Germany).
Deionised water obtained from Millipore MilliQ Academic water
purification system (France) with a resistivity of 18.2 MO was used
throughout the experiments. The multi-element standard stock
solutions were prepared from 1000 mg L1 (High Purity, USA) mix-
ture. The standard solutions were prepared from 10 mg L1 stock
mixture freshly.
2.4. Sample preparation and standard addition
The microwave (MW) digestion system used for the decompo-
sition and digestion of the samples was a Milestone Ethos SEL
(Sorisole, Italy) equipped with MPR 600/12 rotor. 1–1.5 g of sample
was placed into PTFE digestion vessel and 7 mL of nitric acid (65%,
w/w) and 1 mL of hydrogen peroxide (30%, w/w) was added. In
each set of digestion, one reagent blank sample was prepared in
one of the PTFE digestion vessels by just transferring the reagent
mixture (7 mL nitric acid and 1 mL hydrogen peroxide) into this
vessel. The samples and blank sample in closed vessels were sub-
jected to a two-step microwave digestion program. In the first step
of the optimised digestion program, the power of 1000 W was used
and the temperature was increased from room temperature to
200 °C within 10 min. In the second step, the temperature was kept
constant at 200 °C for 20 min and 10 min ventilation time was set
up. Then the digestion vessels were cooled down to room temper-
ature before opening and transferring to volumetric flasks. The
samples and blank sample were diluted to 50 mL with deionised
water using precleaned perfluoroalkoxy (PFA) volumetric flasks,
which were calibrated and certified.
Standard addition method was used for all calibrations. For the
dilution of standard solutions, a representative sample matrix was
prepared by mixing at least 10–15 sample digests. The standards
were prepared in 1:2 diluted sample matrices.
3. Results and discussion
3.1. Validation
The method performance was evaluated in terms of linearity,
limit of detection and quantification, trueness, recovery and preci-
sion. The results obtained for the concentration of Cd, Cu, Fe, and
Zn of NIST SRM 1573a CRM was used to validate the method.
3.1.1. Linearity
Standard addition method was used to obtain calibration
curves. The calibration lines for all the isotopes monitored had a
correlation coefficient of at least 0.994 or better (Table 2). The
calibration curve plot for Cd was shown in Fig. 1. The residual stan-
dard deviation (S) value and calibration data values were also
added to the graph to have an idea about the residuals of the plot.
3.1.2. LOD and LOQ
The limit of detection was estimated as the concentration corre-
sponding to three times the standard deviation (3r) of the blank
signals obtained from a set of reagent blanks (n = 10). The limit
of detection (LOD) and the limit of quantification (LOQ, 10r) for
 S. Yenisoy-Karakasß / Food Chemistry 132 (2012) 1555–1561 1557

Table 1 Table 2
Operating conditions for the HR-ICP-MS. Linear range, LOD and LOQ values.

Parameter Value Elements r (correlation Lineara range LOD LOQ

coefficient) (lg L1) (lg L1) (lg L1)
RF power (W) 1150
Coolant gas flow (L min1) 16 Cd 0.998 0–200 0.18 0.60
Auxiliary gas flow (L min1) 0.8 Cu 0.994 0–200 0.16 0.53
Nebulizer PFA micronebulizer (50 lL min1) Fe 0.998 0–200 0.90 3.00
Sample gas flow (L min1) 1.18 Pb 0.999 0–200 0.19 0.63
Sampling cone (orifice dia., mm) Platinum (1) Sn 0.998 0–200 2.75 9.16
Skimmer cone (orifice dia., mm) Platinum (0.8) Zn 0.997 0–200 0.64 2.14
Injector Sapphire a
Peristaltic pump flow (rpm) 18 Values represent the concentration of standard spiked into sample matrix.
Spray chamber PFA
Nominal resolution LR = 400, MR = 4000, HR = 10000
Scanning mode Both (analog and counting) values for the metals changed between 1.7 and 6.5 except for the
Number of scans for each isotope 9 for each resolution
values for Fe with a value of 16. The %RSD values for the first spik-
Sample time (ms) 10 (LR), 20 (MR), 50 (HR)
Ion lens setting Adjusted to obtain maximum ing level are higher than that of the higher spiking level as
signal/noise ratio expected. Generally, the recovery values for the first level of spik-
Auto sampler CETAC ASX-510 ing are lower than the recovery values for the second level.
Rinse time (s) 20
Sample uptake time (s) 80
3.2. Estimation of uncertainty
LR, low resolution; MR, medium resolution; HR, high resolution.
3.2.1. Identification of uncertainty sources
The element concentrations in the samples expressed in lg g1
the elements determined are shown in Table 2. The detection lim- were calculated by using the Eq. (1):
its for all the elements investigated were found to be in the range
of 0.16 and 2.75 lg L1. CA  V final 1
C¼  DF  ð1Þ
M  1000 R
3.1.3. Trueness where C: concentration of the element in tomato sample (lg g1);
In order to evaluate the trueness of the complete method CA is the element concentration in the sample obtained from the
including digestion and the measurements, NIST SRM 1573a (to- calibration (lg L1); Vfinal is the final diluted volume (50 mL) after
mato leaves) CRM was analysed together with the samples apply- the digestion; M is the mass of a sample (g); DF: Dilution Factor
ing the same digestion and measurement procedure. Each set of (samples are diluted five times before the analysis); R: Recovery.
sample digestion contained one CRM (0.5–0.7 g) which is mea-
sured together with the same sample set. Reagent blank values 3.2.2. Identification of standard uncertainties associated with each
were subtracted from the sample values in the calculation of recov- step
ery values. Bias values for the elements Cd, Cu, Fe and Zn whose The main parameters that affect the concentration of the ele-
values were certified in the CRM are shown in Table 3. The calcu- ments were the mass of sample, the concentrations that were
lated percentage relative error values varied between 1.4% and obtained from the calibration curve, final volume of sample
9.0%. digested, dilution factor, recovery and repeatability. Therefore,
the following parameters were involved in the calculation of the
3.1.4. Recovery and precision concentrations of the elements in the tomato paste samples:
For recovery calculations, the spiking was performed at two for-
tification levels on sample matrix representing different parts of (1) Calibration curves and stock solutions of standards (CA)
the linear range. Five tomato paste samples were spiked with met- (2) Final volume of the sample digested (Vfinal)
als at each fortification level at the same day and analysed in the (3) Mass of the sample (M)
consecutive 5 days. The mean recovery and the precision (the per- (4) Repeatability (Rep)
centage relative standard deviation (%RSD)) data are given in Table (5) Dilution factor (DF)
4. The recoveries ranged from 91% to 103%. The percentage RSD (6) Recovery (R)

y = 112301x + 37721745 S=554642

30000000
r=0.9985
25000000
Intensity (cps)

20000000

15000000

10000000

5000000

0
0 50 100 150 200
Cd Concentration (ug L-1)

Fig. 1. The calibration plot of Cd.

1558 S. Yenisoy-Karakasß / Food Chemistry 132 (2012) 1555–1561

Table 3 3.2.2.2. Estimation of the uncertainty derived from the weighing of the
Percentage relative error obtained for each element using NIST SRM 1573a (tomato sample, u(M). The balance uncertainty is obtained from the certif-
leaves) CRM.
icate given by TUBITAK-UME with the Eq. (5) at 95% confidence
Element Number Experimental Certified % Relative level.
of concentration value ± U (k = 2) error ± STDEV
samples (lg g1) (lg g1) 0:1 þ 1:59  105  w ð5Þ
Cd 4 1.48 1.52 ± 0.04 2.52 ± 0.97
Cu 4 4.74 4.7 ± 0.14 1.74 ± 2.13 where w is the weight of the sample in milligrams. The sample
Fe 4 335 368 ± 7 9.01 ± 1.81 weight was taken as 1500 mg. The uncertainty from this equation
Zn 4 30.7 30.9 ± 0.7 1.39 ± 0.78 was calculated as 0.12385. In order to convert the calibration uncer-
tainty to the standard uncertainty it was divided by 2 and 0.062 was
obtained.
The calibration source of uncertainty was considered twice
Table 4
Comparison of the recovery and repeatability data at two fortification levels.
because the weighing process involves a difference (weighing va-
lue of sample was subtracted from the tare value). The repeatabil-
Elements 1st level 2nd level ity of the balance was included in the above equation. Therefore,
Spike Mean RSD Spike Mean RSD the combined standard uncertainty was calculated by the using
(lg g1) recovery (%) (lg g1) recovery (%) the Eq. (6):
Cd 1.6 91 5.2 3.0 101 2.1 qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Cu 4.0 95 3.0 9.0 96 2.9 uðmsample Þ ¼ ðucalsample Þ2 þ ðucaltare Þ2 ð6Þ
Fe 7.8 96 16 23 99 4.6
Pb 4.0 103 6.5 7.7 103 1.7 It was calculated as 0.0876 mg for a 1500 mg sample.
Sn 11 99 3.2 23 100 1.7
Zn 10 96 4.6 23 95 4.2
3.2.2.3. Estimation of the uncertainty derived from the estimation of
the calibration curves, stock solutions of standards, u(CA). This term
is a combination of the uncertainties associated with the prepara-
Thus the uncertainty in the concentrations is the combination tion of the stock solutions and the calibration curve. However, the
of the uncertainties of each above mentioned parameters. The calculations of uncertainties coming from the calibration curve
combined uncertainty (in terms of relative uncertainty) can be cal- and standard stock solution were performed separately. The uncer-
culated by using the Eq. (2): tainty of preparation of standard stock solution was calculated
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
using the Eq. (7). The uncertainty of the preparation of stock solu-
urel ðCÞ ¼ u2rel ðCAÞ þ u2rel ðV final Þ þ u2rel ðMÞ þ u2rel ðrepÞ þ u2rel ðDFÞ þ u2rel ðRÞ
tions (10 mg L1) is also a combination of uncertainty of elemental
ð2Þ concentrations given in the certificate by the manufacturer, uelement,
and uncertainty coming from the volumetric flasks and pipettes
3.2.2.1. Estimation of the uncertainty derived from the dilution of the used for dilution, uV. The uncertainties of calibration standards
sample digest, u(Vfinal) and dilution factor, u(DF). The uncertainty of are certified as 1000 ± 3 mg L1 for each element. The Eq. (4) was
the dilution depends on the temperature and on the volume mea- used to calculate the uncertainty coming from the volumetric flasks
surement error. After the decomposition procedure the samples as well as the micropipettes.
were diluted to 50 mL with deionised water. qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
The temperature effect describes the dispersion produced by a ustd ¼ ðuV =VÞ2 þ ðuelement =C element Þ2  C std ð7Þ
variation of liquid temperature through a rectangular distribution
Celement: concentration of element in main stock solution
(Eurachem/Citac guide, 2000) within ±3 °C around the calibration
(1000 mg L1), Cstd: concentation of intermediate standard solu-
temperature.
tions (10 mg L1).
3V Q The uncertainty associated with the calibration curve is calcu-
uvtemp ¼ ð3Þ
1:73 lated by using the Eq. (8) given in Eurachem/Citac guide, 2000.
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
where uvtemp represents the standard uncertainty of the tempera- X
S 1 1 ðc0  cÞ2 n
ture effect, V is the measured volume (e.g. 50 mL) and Q is the coef- uðc0 Þ ¼ þ þ Sxx ¼ ðci  cÞ2 ð8Þ
ficient of volume expansion of the water (Qwater = 0.000124 °C1). B1 p n Sxx i¼1
The volumetric calibration standard uncertainty, uvcal, for 5 mL
where S: residual standard deviation, B1: slope, p: number of
automatic pipette, is calculated from the manufacturer reported
measurements to determine c0, n: number of measurements for
CV value of 0.08 (i.e. uvcal = (0.08/100) ⁄ 5 = 0.004 mL). These values
the calibration, c0: determined analyte concentration in tomato
are combined in the Eq. (4) to calculate the uncertainty arising from
paste, c : mean value of the different calibration standards (n num-
the dilution of the sample solutions.
ber of measurements), i: index for the number of measurements to
These sources are combined in the Eq. (4):
obtain the calibration curve, ci: individual calibration standard va-
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
lue obtained from the calibration equation.
uV ¼ ðuvcal Þ2 þ ðuvtemp Þ2 ð4Þ
The five calibration standards were prepared and the instru-
The standard uncertainty of repeatability is not included here ment was run to acquire nine replicate measurements from each
since the repeatability for all possible sources is collected under solution (in this case, p = 9 and n = 45).
repeatability term separately. The uncertainty of the volumetric
flask with 100 mL capacity was ±0.014 mL, the uncertainty of the 3.2.2.4. Estimation of the repeatability uncertainty, u(rep). The uncer-
volumetric flask with 50 mL capacity was ±0.023 mL. The variation tainty from the repeatability covers the contributions from purity
coefficient of an automatic pipette with the range of 0.5 and 5 mL of acids, losses from digestion procedure, accuracy of calibration,
is given as 0.08 and the variation coefficient of an automatic pip- stability of the instrument etc. Whole procedure was applied to
ette with the range of 100 and 1000 lL is given as 0.23 in the five subsamples prepared and analysed in five different days. The
certificates. standard deviation of the repeatability data mentioned above
 S. Yenisoy-Karakasß / Food Chemistry 132 (2012) 1555–1561 1559

represented the repeatability standard uncertainty. To achieve the tcritical for all elements. That means that all Rs values were statisti-
standard uncertainty, mean %RSD of these samples was divided by cally significantly different from 1 and it was convenient to include
pffiffiffi
the square root of the number of samples, n = 5, (%RSD/ n). The the recovery correction factor in the model equation.
variability from the samples was eliminated by combining and Therefore, for the u(R) calculation of the elements Cd, Cu, Fe and
homogenising different digestions. Zn, the dominant factor is (u(Rm)/Rm)2. The absence of this factor in
the calculation of u(R) for the elements Pb and Sn caused underes-
3.2.2.5. Estimation of the recovery uncertainty, u(R). The overall R is timation of u(R) values for the elements Pb and Sn.
calculated by considering the contribution from the method Rm However, in the routine application of the method, the differ-
and the contribution from the tested sample Rs. Rm represents a ence is not considered to be of practical significance and no correc-
test of bias against a particular reference value, it is also necessary tion to the final result is applied (D’llio, Alessandrelli, Cresti, Forte &
to consider differences between it and the recovery for real sam- Caroli, 2002). In this case, the uncertainty associated with method
ples. Rs represents this difference between the reference and a recovery must be increased to include this uncorrected bias using
particular sample (Barwick et al., 1999). The overall recovery, R, the Eq. (15):
is calculated by the Eq. (9) as follows: sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
  2
 ¼ 00 1R  2
R ¼ Rm  Rs ð9Þ uðRÞ þ uðRÞ ð15Þ
k
The method recovery, Rm, for a particular sample can be consid- where k is the coverage factor that will be used in the calculation of
ered as Cobserved/CCRM. Rm is an estimate of the recovery for the expanded uncertainty.
entire procedure, including digestion and analysis. Rm is measured
(ideally) with a suitable certified reference sample. This value has
3.3. Estimation of expanded uncertainties
an uncertainty associated with the reference value used and with
the variability of the particular measurement of the recovery on
As an example, the calculation of relative uncertainty for ele-
that material. The standard uncertainty of Rm is calculated by using
ment Cd is given in Table 5. The relative expanded uncertainties
the Eq. (10):
(%) of elements Cd, Cu, Fe, Pb, Sn and Zn were 13.8, 13.9, 14.0,
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 2 8.20, 13.2 and 16.4, respectively. The percentage contribution of
s2obs uðC CRM Þ
uðRm Þ ¼ Rm þ ð10Þ each source to the relative combined uncertainties is shown in
n  C 2m C CRM Fig. 2. It might be seen that the contribution from the calibration
This calculation was performed for the elements, Cd, Cu, Fe and curve, repeatability and recovery were the major sources. The
Zn, because NIST SRM 1573a (tomato leaves) has certified concen- uncertainties coming from the mass of sample, preparation of
tration for these elements. For the elements Pb and Sn, the recov- stock solutions (contributes to the concentration obtained from
ery values obtained from the spiking experiments were used. If the the calibration curve, CA), dilution after digestion and dilution
spiked samples are prepared from a sample matrix which contains required for the analysis did not contribute very much to the rela-
the analyte, the situation is complex (Barwick & Ellison, 2000). The tive combined uncertainties.
recovery for each sample, Rs(i), is given by the Eq. (11):
3.4. Analysis of real samples
C obsðiÞ  C native
RsðiÞ ¼ ð11Þ
C spikeðiÞ The tomato pastes obtained from the supermarkets were ana-
lysed using the validated method. Number of samples, arithmetic
where spike is pure substance which is added to a tomato paste mean, standard deviation, minimum and maximum values were
matrix/sample. presented in Table 6. The concentrations of elements Cu, Zn and
The simplified form of the standard uncertainty of recovery is Fe were previously determined in various types of canned foods
calculated by the Eq. (12). marketed in Turkey (Tüzen & Soylak, 2007a). The observed metal
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi concentrations in Turkish tomato paste samples are comparable
 ðC obsðiÞ Þ2
u
sÞ ¼ 1
uðR þ uðC native Þ2 ð12Þ
with the values in that study (Tüzen & Soylak, 2007a) and also
 spike
C n studies cited in the paper. For example, the lowest average value
for Cu was 2.85 lg g1 (canned pickle) and the highest average
The uncertainty associated with R, u(R) is given by the Eq. (13): value was 7.77 lg g1 (canned bean). The lowest and the highest
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi average values for Zn were found as 8.46 lg g1 in canned pickle
uðRm Þ 2 uðRs Þ 2 and 21.9 lg g1 in canned mushroom samples. Iron content ranged
uðRÞ ¼ R  ð Þ þð Þ ð13Þ
Rm Rs from 27.5 lg g1 in canned delicatessen to 79.6 lg g1 in canned
A significance test was used to determine whether the R value
was significantly different from 1.0. If the resulting R value is sta- Table 5
tistically different from 1 after applying the significance test, the Calculation of relative uncertainty (%) for Cd determination in tomato paste samples.
recovery correction factor should certainly appear in the model Parameter Value (X) u(X) u(X)/X
equation. The test statistic t, was used to perform the significance
Stock solution (mg L1) 10 0.028 0.00280
test and calculated by the Eq. (14): Sample mass (mg) 1500 0.0876 0.0000584
Sample volume (mL) 50 0.0293 0.000590
j1  Rj Sample dilution 5 0.0010 0.000207
t¼ ð14Þ
uðRÞ Calibration uncertainty (lg L1) 50 2.384 0.0477
Recovery 1 0.0468 0.0468
The value from this equation was compared with two-tailed Repeatability 100 1.753 0.0175
critical values at 95% confidence level. According to the test, the Measurement result (lg g1) 0.0691
Rm value (Table 3) for Fe was statistically significantly different Standard combined uncertainty 0.00484
Expanded uncertainty (k = 2) 0.00978
from 1. The Rm values were taken as 1 for Cd, Cu and Zn. Using
Relative uncertainty (%) 13.8
the Rs values (first level) (Table 4), the calculated t was greater than
1560 S. Yenisoy-Karakasß / Food Chemistry 132 (2012) 1555–1561

Fig. 2. Percentage contribution of each effect to the total uncertainty for tomato paste samples.

elements, by high resolution ICP-MS. The method was successfully

Table 6
Statistical overview of analysed tomato samples gathered from the supermarkets in validated with the parameters such as trueness, precision, recov-
Turkey (lg g1). ery, LOD, LOQ and linearity.
Using the approach of the bottom up method, dilution factor,
Elements N Arithmetic average Standard deviation Min. Max.
final volume, mass of the sample, calibration curve, standard solu-
Cd 14 0.070 0.040 0.023 0.17
tion, recovery and repeatability were found to be the sources of
Cu 14 2.56 0.651 1.57 3.37
Fe 14 22.5 11.1 11.4 57.5
uncertainty. In the calculation of uncertainty, it was seen that
Pb 14 0.045 0.026 0.019 0.096 the calibration curve, recovery and repeatability have the domi-
Sn 14 1.94 1.85 0.030 6.04 nant effects on the combined uncertainty. The percentage relative
Zn 14 4.91 1.11 3.57 7.72 expanded uncertainties for each element using coverage factor 2
were between 8.2% and 16.4%.
mushroom samples (Tüzen & Soylak, 2007a). However, the maxi- The validated method was used to analyse 14 different trade
mum iron level permitted for canned food is 15 lg g1 according mark canned tomato pastes that were purchased from the super-
to Turkish Food Codex (Turkish Official Gazette, 2002). Iron levels markets. The concentrations of Cd and Fe exceeded the limits of
in analysed canned tomato paste samples were found to be higher European Commission Regulations and Turkish Food Codex,
than the legal limits. This may be due to processing of raw materi- respectively.
als in iron and steel vessels.
As a result of the use of tinplate for food and beverage packag-
Acknowledgements
ing, it is obvious that some tin will dissolve into the food content,
particularly when plain internal (unlacquered) surfaces are used
The author would like to thank to Ibrahim Akdağ and Dr. Oktay
(Blunden & Wallace, 2003). The recommended maximum permis-
Cankur for their assistance in sample preparation and analysis. The
sible levels of tin, lead and cadmium in food are 200, 0.1 and
author is grateful to Dr. Nur Banu Öztasß for her valuable
0.05 mg kg1, respectively in European Commission Regulations
suggestions.
(2006). While lead concentrations came close to the maximum
permissible level of European Commission Regulations, cadmium
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