Accred Qual Assur (2013) 18:235–241
DOI 10.1007/s00769-013-0979-6
PRACTIONER’S REPORT
Evaluation of measurement uncertainty components associated
with the results of complexometric determination of calcium
in ceramic raw materials using EDTA
Samarendra Basak • Dipali Kundu
Received: 23 July 2012 / Accepted: 3 April 2013 / Published online: 24 April 2013
Ó Springer-Verlag Berlin Heidelberg 2013
Abstract Calcium is an important constituent of mineral
like calcite, dolomite, gypsum and bio-ceramic raw mate-
rial like hydroxyapatite. Those are frequently used for the
manufacture of cement, mortar, glass, synthetic ceramic
bone supplement, dental enamel, etc. Determination of
exact quantity of calcium in those materials is therefore
very essential. The calcium content has been determined
complexometrically in a ceramic raw material at pH 12,
using di-sodium salt of EDTA. The major sources of
uncertainty of the results of measurement are contributions
from repeatability, standardization of EDTA, volume
measurement by volumetric flask, burette, pipette and end
point detection. Sources of uncertainty have been identified
and combined by following the EURACHEM guidelines.
The results show that the major sources of uncertainty arise
from standardization, repeatability of the experiment and
end point detection by burette. Cause–effect diagram has
been drawn to explain the uncertainty budget.
Keywords Uncertainty measurements  Calcium 
Apatite  Gypsum  Calcite  Lime stone
Introduction
Calcium occurs abundantly in sedimentary rocks in the
minerals calcite, dolomite, gypsum, etc. Calcium carbonate
is used in the manufacture of cement, mortar, lime and glass.
Moreover, antacids frequently contain calcium carbonate
and are commonly used inexpensive calcium supplement.
S. Basak (&)  D. Kundu
Analytical Chemistry Section, Central Glass and Ceramic
Research Institute, 196, Raja S.C Mullick Road,
Kolkata 700 032, India
e-mail: sbasak@cgcri.res.in
Hydroxyapatite usually written as Ca10(PO4)6(OH)2 makes up
70 % of bone. Carbonated calcium deficient hydroxyapatite is
the main mineral constituting the dental enamel and denting.
Bio-ceramics based on calcium phosphate includes hydroxy-
apatite, b-tricalcium phosphate and there composite materials
are bio-compatible and are widely used as bone replacement
materials in the fields of orthopaedic, oral and plastic surgery
[1]. Studies [2] have shown that the sintering stability,
mechanical strengths, thermal stability, degradability and
biological properties of calcium phosphate-based bio-ceramics
are largely affected by the chemical composition of Ca/P ratio.
In all these cases, accurate determination of calcium con-
tent is very essential for quality control of the desired products.
Determination of calcium in samples is usually carried out by
atomic absorption spectroscopy [3] and inductively coupled
plasma atomic emission spectrometry [4]. For the determi-
nation of higher quantity of calcium, it is not appropriate to use
instrumental methods due to large dilution errors. In such case,
complexometry is very widely used using di-sodium salt of
EDTA as complexing agent [5, 6].
The exact value of a quantity is indeterminate. The mea-
surement process, whatever carefully planned and conducted,
there remains uncertainty about the true value of the quantity
being measured. The results of measurements are therefore
incomplete without a realistic statement of its uncertainty.
Often the uncertainty of the result is calculated considering the
standard deviation of repeated determinations. In this way, the
uncertainty is underestimated because only uncertainty com-
ponents arising from random effects (repeatability) are
considered. The overall analytical uncertainty is sometimes
much larger and contains uncertainty components arising
from systematic effects. The EURACHEM guide for
‘‘Quantifying Uncertainty in Analytical Measurement’’ [7] is
based on the ‘‘Guide to the expression of uncertainty in
measurement’’ published by ISO [8]. According to the guide,
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the uncertainty may be estimated from its components by
using the rules for propagation of errors in order to combine
them into total uncertainty. The uncertainty budget considered
in this study is based on these guidelines.
The estimation of uncertainty requires the analyst to
look closely at all the possible sources of uncertainty [9].
Measurement uncertainty evaluation of the results of
measurement is one of the few requirements of the standard
ISO/IEC 17025 [10] which must be fulfilled by the labo-
ratory in order to obtain accreditation for certain test
methods. Very few studies on measurement uncertainty
have so far been reported [11–20].
In the present study, mass fraction of CaO in the apatite
concentrate has been determined by complexometric
method. Calcium present in samples is determined by
titration with EDTA at pH 12 using fluorexone (calcein) as
indicator. As per GUM/EURACHEM guidelines, major
sources of uncertainties have been identified, uncertainties
at each step have been calculated and have been included
in the calculation of combined uncertainty.
Experimental
Reagents and apparatus: the reagents, di-sodium salt of
EDTA, concentrated hydrochloric acid (HCl), concentrated
nitric acid (HNO3), sodium hydroxide (NaOH), etc. used
are all of AR/GR grade from E. Merck (Germany). Indi-
cator is prepared by thoroughly mixing 0.2 g of fluorexone
with 20 g potassium chloride and about 0.1 g of this
mixture is used for each titration. Volumetric standard
calcium carbonate (E. Merck, Germany) is used for stan-
dardization of EDTA solution. Burette, pipette and
volumetric flask used are all of class A grade, and double
distilled water is used in all the cases.
Description of the method: Concentration of calcium is
determined according to the standard method [5, 6]. EDTA
solution of about 0.025 mol/L strength is prepared as usual.
For standardization of EDTA, 0.0625 g of volumetric
standard calcium carbonate, dissolved in 10 % HCl solu-
tion is directly titrated with EDTA solution at pH 12 using
fluorexone indicator. The sample solution is prepared by
using hydrochloric acid and hydrofluoric acid, and final
volume is made 100 mL in a volumetric flask.
Results and discussion
Specification of the measurand and identification
of the sources of uncertainty
The main sources of uncertainty which are considered in
the measurement are identified as the repeatability of the
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Accred Qual Assur (2013) 18:235–241
experiment, measurement of strength of EDTA, volume
measurement by volumetric flask, burette and pipette,
measurement of mass and reagent purity. The sources of
uncertainty are schematically presented in Figs. 1 and 2
with serial numbers (#1–#13) on each parameter, and those
serial numbers are also included in the text. Repeatability
contributions to uncertainty are combined into one contri-
bution for the overall experiment, and values are obtained
from the method validation study [5, 6]. The uncertainty
associated with the digestion of the sample and homoge-
neity (#10) are assumed to be negligible. The mass fraction
of CaO, WCaO, in the sample (#9) is expressed by the
model:
0
VEDTA SEDTA VSOL FREP2
WCaO ¼ ð1Þ
VALQ m2
where WCaO denotes mass fraction of CaO, VEDTA 0
the
volume of EDTA consumed for sample, SEDTA the CaO-
equivalent concentration of EDTA, VSOL the volume of
solution prepared for sample, VALQ the volume of aliquot
taken for sample, m2 the mass of sample and FREP2 the
repeatability of the overall measurement of mass content
CaO.
SEDTA is calculated from the equation:
m1 P f FREP1
SEDTA ¼ ð2Þ
VEDTA
where m1 denotes the mass of calcium carbonate, P the
purity of calcium carbonate in g/(100 g), f the molecular
weight ratio of CaCO3/CaO, FREP1 the repeatability of
standardization of EDTA and VEDTA the volume of EDTA
consumed for standardization.
Linearity (L) of the balance is the maximum difference
between the actual mass on the pan and the reading of the
scale. Balance manufacturer’s own uncertainty evaluation
recommends the use of a rectangular distribution to convert
the linearity contribution to a standard uncertainty. The
distribution has to be counted twice, once for tare and once
for gross weight, because each is an independent obser-
vation and the linearity effects are not correlated.
Therefore, standard uncertainty of mass m is:
pffiffiffi pffiffiffi
uðmÞ ¼ 2L= 3 ð3Þ
The uncertainty associated with the measurement of
volume of EDTA depends on the uncertainty of the volume
of the burette itself, the uncertainty associated with the use
of the burette at a temperature different from that of the
calibration temperature and the repeatability of the volume
delivery. In this case, limits of accuracy of the measurement
of volume are indicated by the manufacturer with no
information on distribution. The ISO guide recommends [8]
a triangular distribution assumption. Therefore, to obtain
Accred Qual Assur (2013) 18:235–241 237

Fig. 1 Cause-effect diagram for standardization of EDTA by standard CaCO3 (symbols are explained in text and Table 1)

Fig. 2 Cause-effect diagram for estimation of uncertainty of mass fraction of CaO in a sample (symbols are explained in text and Table 1)

the standard uncertainty, the values are divided by H6. coefficient of water (c) lead to a volume variation calculated
Volume uncertainty due to temperature variation, u(VT) is as:
calculated from an estimate of the temperature range and VDTc
the volume expansion coefficient. A temperature variation uðVT Þ ¼ pffiffiffi ð4Þ
3
of ±3 °C is taken as reasonable estimate (with a 95 %
confidence). The volume expansion of water is considerably Error due to end point detection has a rectangular
larger than that of glass, so only the former is considered. distribution, and the corresponding uncertainty may be
The temperature variation DT and the volume expansion calculated as:

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0:5Si
uðVEND Þ ¼ pffiffiffi
3
where Si denotes scale increment in volume/graduation of
the concerned burette.
All uncertainty components are then combined to obtain
the uncertainty in volume measurement by the burette
u(VBUR) as:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðVBUR Þ ¼ uðVCAL Þ2 þuðVT Þ2 þuðVEND Þ2
where, u(VCAL) is the uncertainty associated with the cal-
ibration of the burette or volumetric flask or pipette (as the
case may be) which are obtained from producer’s certifi-
cate and u(VT) is the uncertainty associated with the use of
the burette or volumetric flask or pipette (as the case may
be) at a temperature different from that of calibration
temperature.
The uncertainty associated with measurement of volume
by volumetric flask, u(VSOL) is obtained following the
equation:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðVVOL Þ ¼ uðVCAL Þ2 þuðVT Þ2 ð7Þ
Similarly, the uncertainty associated with measurement
of volume by pipette, u(VPIP) is obtained following the
equation:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðVPIP Þ ¼ uðVCAL Þ2 þuðTT Þ2 ð8Þ
Calculation of the combined and expanded uncertainty
The uncertainty associated with the measurement of cal-
cium content in the sample is estimated by combining the
standard uncertainties of all the components under con-
sideration. To calculate the expanded uncertainty of the
result of measurement at 95 % confidence level, the result
of combined uncertainty is multiplied by a coverage factor
of k = 2. Relative uncertainty variance contributions are
used to illustrate the relative impact of different uncertainty
components.
The complexometric method for the determination of
calcium using EDTA involves two main steps:
Step A: Quantification of uncertainty components
for EDTA standardization
Here, tolerance values for class A burette (IS 1997:1982) of
25 mL capacity is ±0.05 mL, and for 25 mL pipette (IS
1117:1975), it is ±0.03 mL. For 100-mL volumetric flask
(IS 915:1975), the corresponding value is ±0.10 mL.
Repeatability (#4): Our in-house study shows a relative
standard deviation (RSD) of repeatability value of 0.20 %
for the standardization of EDTA by CaCO3 (CRM). This
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Accred Qual Assur (2013) 18:235–241
value has been directly used for the calculation of standard
ð5Þ uncertainty.
Mass measurement (#5): Analytical balance used is
Mettler Tolledo AG 245 with deviation from linearity by
up to ±0.15 mg. Therefore, balance linearity contribution
is 0.15 mg/H3 = 0.0866 mg. So using Eq. 3 standard
uncertainty for the measurement of mass of calcium car-
bonate is u(m1) = 0.000122 g.
Purity of calcium carbonate (#6): The uncertainty
ð6Þ
associated with the purity (P) of the reagent is calculated
from the producer’s specification. Volumetric standard
CaCO3 (E. Merck, Germany) having the assay
(99.94 ± 0.05) % has been used for standardization.
Assuming rectangular distribution uncertainty associated
with purity of CaCO3 is u(P) = 0.05/H3 = 0.028867.
Molecular weight of CaO and CaCO3: Standard uncer-
tainty of the molecular weight u(MCaO) is calculated from
the uncertainties of MCa and MO by using the formula as :
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðMCaO Þ ¼ uðMCa Þ2 þuðMC Þ2 þ3uðMO Þ2 ¼ 0:002374:
and standard uncertainty of the molecular weight
u(MCaCO3 ) is calculated from the uncertainty values of
MCa, MC and MO by using the formula as:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðMCaCO3 Þ ¼ uðMCa Þ2 þuðMC Þ2 þ3uðMO Þ2 ¼ 0:002374:
The uncertainties and distribution corresponding to the
atomic weights of concerned elements are obtained from
literature [21], and the corresponding uncertainty of factor
f (#1) may be calculated as:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uðf Þ ¼ uðMCaO Þ2 þuðMCaCO3 Þ2 ¼ 0:003342:
Measurement by burette (#2): data corresponding to
uncertainty associated with measurement of volume by
burette (25 mL) are shown in Table 2, wherefrom using
Eq. 6, we get the uncertainty of the EDTA volume,
u(VEDTA) = 0.036478 mL. All the above calculated values
are tabulated in Tables 1 and 2. Considering the data from the
Table 1 and using Eq. 2, we get SEDTA = 0.001417 g/mL; the
quantity is listed as #3 in Fig. 2.
Now combining the standard uncertainties of all the
components under consideration combined uncertainty
uc(SEDTA) may be obtained by using the formula as:
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
     ffi
uc ðSEDTA Þ uðm1 Þ 2 u ð PÞ 2 uð f Þ 2
¼ þ þ
SEDTA m1 P f
 2  2
uðFREP1 Þ uðVEDTA Þ
þ þ ¼ 0:006764
FREP1 VEDTA
ð9Þ
Therefore, uc(SEDTA) = 0.0000096 g/mL.
Accred Qual Assur (2013) 18:235–241 239

Step B: Quantification of uncertainty components
for sample analysis
Repeatability (#11): The method validation study [5, 6]
shows a relative SD (RSD) of 0.56 % for the overall pro-
cess of measurement of calcium by EDTA. This value can
be directly used for the calculation of standard uncertainty.
shown in Table 2, and from those data using Eq. 7, we get
u(VSOL) = 0.054678 mL.
Pipette (#13): Data for uncertainty due to pipette of
25 mL capacity are shown in Table 2, and here using
Eq. 7, we get u(VALQ) = 0.015254 mL.
Analytical balance (#12): Same analytical balance is
also used here, and using Eq. 3, corresponding standard

Table 1 Uncertainty components and their relative standard uncertainties

Uncertainty Description Value (x) Standard Relative standard
components uncertainty uncertainty u(x)/x
u(x)

FREP1 (Step A) Repeatability for standardization 1.0 0.0020 0.0020

FREP2 Repeatability for overall measurement of Ca 1.0 0.0056 0.0056
m1 Mass of calcium carbonate in g 0.0625 0.000122 0.001952
m2 Mass of sample in g 0.2006 0.000122 0.000608
P Purity of calcium carbonate in g/100 g 99.94 0.028867 0.000289
f Molecular weight ratio of WCaO/MCaCO3 0.560287 0.003342 0.005965
VEDTA Volume of EDTA consumed for standardization in mL 24.70 0.036478 0.001477
0
VEDTA Volume of EDTA consumed for sample in mL 16.20 0.035842 0.002212
VSOL Volume of solution prepared for sample in mL 100 0.051565 0.000516
VALQ Volume of aliquot taken for sample in mL 25 0.015254 0.000610
SEDTA CaO-equivalent concentration of EDTA (in g/mL) 0.001417 0.000010 0.007057

Table 2 Uncertainty associated with the measurement by burette, volumetric flask and pipette
Uncertainty Value Distribution Divisor Standard uncertainty

For burette (25 mL)

Calibration ±0.05 Triangular H6 0.020412
Temperature variation (standardization) 24.70 9 3 9 2.1 9 10-4 Rectangular H3 0.008984
Temperature variation (sample) 16.20 9 3 9 2.1 9 10-4 Rectangular H3 0.005892
End point variation ±0.05 Rectangular H3 0.028867
For volumetric flask (100 mL)
Calibration ±0.10 Triangular H6 0.040825
-4
Temperature variation 100 9 3 9 2.1 9 10 Rectangular H3 0.036373
For pipette (25 mL)
Calibration ±0.03 Triangular H6 0.012247
Temperature variation 25 9 3 9 2.1 9 10-4 Rectangular H3 0.009093

Burette (#7): Data corresponding to uncertainty associated
with measurement by burette (25 mL) are shown in Table 2,
wherefrom using Eq. 6, we get the uncertainty due to volume
0
measurement of EDTA, u(VEDTA ) = 0.035842 mL.
Volumetric flask (#8): Data corresponding to uncer-
tainty due to volumetric flask of 100 mL capacity are
uncertainty for the measurement of mass of sample is
u(m2) = 0.000122 g. All the above calculated values
are tabulated in Tables 1 and 2. Considering the data from
Table 1 and using Eq. 1, we get WCaO = 45.77 g/(100 g).
For a multiplicative expression, the standard uncertain-
ties are used as follows:

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sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
    0  2 
uðWCaO Þ uðm2 Þ 2 u VEDTA
¼ þ
WCaO m2 0
VEDTA
¼ 0:009121
Therefore, uc(WCaO) = 0.42 g/(100 g) and by multi-
plying with coverage factor (k = 2) we get expanded
uncertainty U(WCaO) = 0.84 g/(100 g).
The values of different uncertainty components with
their standard uncertainty and relative standard uncertainty
values for the sample are given in Table 1. Relative con-
tributions of uncertainty components are graphically
presented in Fig. 3. Therefore, the result of measurement
may be represented as WCaO = (45.77 ± 0.84) g/(100 g).
It is evident from the Fig. 3 that the largest contributions
0
are due to u(SEDTA), u(FREP2) and u(VEDTA ). Calculations
in respect of apatite concentrate are shown here, and sim-
ilar calculation may be done for gypsum, limestone,
calcite, etc. using proper repeatability values.
In the present study, almost all the important uncertainty
sources have been included in the calculation of combined
measurement uncertainty. In routine calculation of mea-
surement uncertainty, all the components smaller than one-
third of the largest one have no significant influence on the
final result and can be neglected. Here, almost all the
sources have been taken into consideration in order to asses
the implication of each of the uncertainty components and
their influence on the final results.
Fig. 3 Comparison of the total uncertainty in the mass fraction
(WCaO) and the parameters that contribute to this uncertainty
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Accred Qual Assur (2013) 18:235–241
ffi
      
uðVSOL Þ 2 uðFREP2 Þ 2 uðVALQ Þ 2 uðSEDTA Þ 2
þ þ þ þ
VSOL FREP2 VALQ SEDTA ð10Þ
Conclusion
The best way for improving the performance of a mea-
surement is the detection of the major uncertainty
components. Systematic uncertainty budget facilitates the
uncertainty evaluation process, and as a result, different
uncertainty components can be easily compared. In this
case standardization, repeatability of the experiment and
end point detection are the target of operation for reducing
the measurement uncertainty.
Acknowledgments The authors are thankful to Prof. I. Manna,
Director of the Institute for his kind permission to publish this paper.
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