Trends
7, 2004
Trends in quality in the analytical
laboratory. I. Traceability and
measurement uncertainty of
analytical results
Isabel Taverniers , Erik Van Bockstaele, Marc De Loose
Credibility of analytical data has never caught the public’s
eye more than today. The key principle for quality and reli-
ability of results is comparability between laboratories and on
a wider, international basis. In order to be comparable,
analytical results must be reported with a statement of
measurement uncertainty (MU) and they must be traceable to
common primary references. This work focuses on traceabil-
ity and uncertainty of results. We discuss different
approaches to establishing traceability and evaluating MU.
We place both concepts in the broader context of analytical
method validation and quality assurance. We give up-to-date
information in the framework of new, more exacting
European and international standards, such as those from
Eurachem/CITAC, IUPAC and ISO.
ª 2004 Published by Elsevier B.V.
Keywords: Analytical method validation; Measurement uncertainty;
Quality assurance; Reliability of results; Traceability
Abbreviations: AOAC, Association of Official Analytical Chemists;
AQA, analytical quality assurance; CCMAS, Codex Committee on
Methods of Analysis and Sampling; CITAC, Cooperation on Interna-
tional Traceability in Analytical Chemistry; CRM, certified reference
material; EAL, European Cooperation for Accreditation; FAO, Food
and Agricultural Organization; IEC, International Electrotechnical
Isabel Taverniers*, Marc De Loose
Department for Plant Genetics and Breeding (DvP), Centre for Agricultural
Research (CLO), Ministry of the Flemish Community, Caritasstraat 21
B-9090 Melle, Belgium
Erik Van Bockstaele
Department for Plant Genetics and Breeding (DvP), Centre for Agricultural
Research (CLO), Ministry of the Flemish Community, Caritasstraat 21
B-9090 Melle, Belgium
Department for Plant Production, Ghent University, Coupure Links 653
B-9000 Gent, Belgium
*Corresponding author.
Tel.: +32-9-272-2876; Fax: +32-9-272-2901;
E-mail: i.taverniers@clo.fgov.be
480 0165-9936/$ - see front matter ª 2004 Published by Elsevier B.V. doi:10.1016/S0165-9936(04)00733-2
Trends in Analytical Chemistry, Vol. 23, No.
Commission; ILAC, International Laboratory Accreditation Coopera-
tion; IQC, Internal quality control; ISO, International Standardization
Organization; IUPAC, International Union of Pure and Applied
Chemistry; LGC, Laboratory of the Government’s Chemists; MU,
measurement uncertainty; QA, quality assurance; QC, quality control;
RM, reference material; SI, Syst
eme International; U(u), Expanded
uncertainty (individual uncertainty factor); WHO, World Health
Organization; X, concentration or amount of measurand
1. Introduction: quality of analytical results
Innumerable types of analytical methods exist in the fields of
analytical and bioanalytical chemistry, biochemistry,
biology, clinical biology and pharmacology and related
application domains, such as forensic, toxicological,
environmental, agricultural and food analyses. Regardless of
the type of method, the scope and the application,
laboratories must be able to produce reliable data when
performing analytical tests for a client or for regulatory
purposes.
With the fast development of analytical methodolo-
gies, great importance is nowadays attached to the
‘‘quality’’ of the measurement data. Quality of analytical
measurement data encompasses two essential criteria –
utility and reliability (Fig. 1) [1]. Utility means that
analytical results must allow reliable decision making.
A key aspect of reliability or validity of results is that
they are comparable, whatever their origin. Compara-
bility between results in the strict sense is provided by
traceability to appropriate standards. Traceability to
common reference standards underlies the possibility of
making a comparison – i.e., a distinction – between
different results. If results are also to be compared in
terms of their quantities or levels of analyte, additional
information on the analytical result is needed – MU.
Uncertainty of results arises from the combination of all
uncertainties of the reference values (to which the
Trends in Analytical Chemistry, Vol. 23, No. 7, 2004
quality
utility
reliability
(validity)
comparability
1. Are 2 results
traceability
comparable to, i.e.
distinguishable from
each other?
Y1 = Y2 ? calibration
Figure 1. Relationship between quality, traceability and measurement uncertainty (MU) of results [3].
results are traceable) and all additional uncertainties
associated with the measurement procedure. MU and
traceability are related concepts, both defining the
quality of analytical data (Fig. 1) [2,3].
Quality of results reflects adequacy (or inadequacy) of
a method in terms of the extent to which the method
fulfils its requirements or is fit for its particular analytical
purpose (see Section 2 below). Quality is always a rela-
tive notion, referring to the requirements fixed before-
hand on the basis of national or international
regulations or customer needs [1,4].
The need for reliability of analytical data is stressed by
the fact that measurement results will be used and may
form the basis for decision making. Unreliable results
bring a high risk of incorrect decisions and may lead to
higher costs, health risks, and illegal practices. Imagine,
for example, the consequences if results are false posi-
tives or if the uncertainty is much larger than reported
[1,5,6].
2. The role of method validation in traceability
and MU
An analysis is a complex multistage investigation of the
values of the properties of materials, i.e., the identity and
the concentration of a specific component in a specific
sample material [2,7]. van Zoonen et al. [1] presented
chemical analysis as a cyclic process in which the final
objective is the generation of chemical information. This
integrated process starts with defining the basic analyt-
ical problem (specifying the analytical requirement) and
ends with evaluating and reporting the analytical result.
Ideally, the last step provides an answer to the initial
problem, as stated by a client or based on regulatory
requirements.
Trends
2. Which of the 2 results
MU estimation
contains, with a certain level
of confidence, the highest
level of analyte?
Y1>Y2 or Y1<Y2 ?
The process of providing an answer to a particular
analytical problem is presented in Fig. 2. The analytical
system – which is ‘a defined method protocol, applicable
to a specified type of test material and to a defined con-
centration rate of the analyte’ – must be ‘fit for a partic-
ular analytical purpose’ [4]. This analytical purpose
reflects the achievement of analytical results with an
acceptable standard of accuracy. Without a statement of
uncertainty, a result cannot be interpreted and, as such,
has no value [8]. A result must be expressed with its
expanded uncertainty, which, in general, represents a
95% confidence interval around the result. The proba-
bility that the mean measurement value is included in the
expanded uncertainty is 95%, provided that it is an
unbiased value that is made traceable to an interna-
tionally recognized reference or standard. In this way, the
establishment of traceability and the calculation of MU
are linked to each other. Before MU is estimated, it must
be demonstrated that the result is traceable to a reference
ANALYTICAL SYSTEM
measurand
VALIDATION ANALYTICAL RESULT VALIDATION
measurement
± uncertainty
value
fitness-for-purpose
INTERPRETATION & EVALUATION
Figure 2. Role of method validation in quality of analytical
measurements. Validation is the process to demonstrate the fitness-
for-purpose of the analytical system [4,8,14,15].
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Trends
or standard which is assumed to represent the truth
[9,10].
Traceability and MU both form parts of the purpose of
an analytical method. Validation plays an important role
here, in the sense that it ‘confirms the fitness-for-purpose
of a particular analytical method’ [4]. The ISO definition
of validation is ‘confirmation by examination and
provision of objective evidence that the particular re-
quirements of a specified intended use are fulfilled’ [7].
Validation is the tool used to demonstrate that a specific
analytical method measures what it is intended to
measure, and thus is suitable for its intended purpose
[2,11]. In part II of this review, the classical method-
validation approach is described, based on evaluation of a
number of method-performance parameters. Summa-
rized, the criteria-based validation process consists of
precision and bias studies, a check for specificity/
selectivity, a linearity check, robustness studies and,
eventually, based on the practical requirements of the
method, an assessment of the limits of detection and/or
quantification.
The objective of validation is to verify that the
measurement conditions and the equation used to
calculate the final result include all the influences that
will affect the final result. Validation measures the dif-
ferent effects, throughout the whole analytical system,
that influence the result, and ensures that there are no
other effects that have to be taken into account. A
specificity test ensures that the method responds to the
specific analyte of interest only, and not to other interf-
erents or contaminants. A linearity check verifies that
the supposed relationship between the signal and units
used for the analyte may be used. A bias study is a
certified reference material (CRM) check that demon-
strates that the method is not significantly biased; and,
precision and robustness studies cover the effects of
variability in conditions, operators, equipment and time.
The role of method validation in the achievement of
reliable results is:
(1) to include all possible effects or factors of influence
on the final result;
(2) to make them traceable to stated references (refer-
ence methods, RMs or SI units);
(3) to know the uncertainties associated with each of
these effects and with the references.
Validation is thus a tool to establish traceability to
these references [2–4]. In this context, it is important to
see the difference between traceability and accuracy. A
method that is accurate, in terms of ‘true’ (i.e., approx-
imating the ‘true value’), is always traceable to what is
considered to be the true value. However, the opposite is
not correct. A method that is traceable to a stated ref-
erence is not necessarily true (accurate). Errors can still
occur in this method, depending on the reference [12].
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Trends in Analytical Chemistry, Vol. 23, No. 7, 2004
Analytical method validation forms the first level of
quality assurance (QA) in the laboratory. Analytical QA
(AQA) is the complete set of measures a laboratory must
undertake to ensure that it is able to achieve high quality
data continuously. Besides the use of validation and/or
standardized methods, these measures are effective
internal quality control (IQC) procedures (use of RMs,
control charts,. . .), participation in proficiency testing
schemes and accreditation to an international standard,
normally ISO/IEC 17025 [4]. Method validation and the
different aspects of QA form the subject of part II of this
review (Method validation and AQA).
3. Guidelines on traceability and uncertainty
of results
Table 1 shows an overview of prominent institutions
offering guidance and their guidelines on traceability,
MU and related topics. In Europe, a leading role is
played by Eurachem, a working group on analytical
chemistry centralized at and originating from the UK’s
LGC (formerly Laboratory of the Government Chemist).
Basic references are CITAC/Eurachem guides on
‘Quality in Analytical Chemistry’ [2] and ‘Traceability
in chemical measurement’ [3], and a Eurachem guide
on MU [13,14]. Eurachem has also published guides
on related topics, such as RMs [7] and method
validation [15].
At the international level, relevant standards are
available from IUPAC, ISO and AOAC International
[4,8,16,17] and from the Codex Alimentarius’s working
group, CCMAS [18–21]. Other helpful guides have been
published by EAL [22] and ILAC [23] (see Table 1 for
explanations of abbreviations).
4. The concept of traceability
4.1. Definitions
Traceability is a relatively new term, gaining more and
more attention in analytical measurement sciences.
Traceability can be assigned to different aspects related
to a measurement – such as traceability of a result,
method, procedure, laboratory, product, material, and
equipment. As such, there is no single definition of
traceability.
Before exploring the different concepts of traceability,
we can look to a more general, extended meaning.
According to Valcarcel and Rios [24], the basic meaning
of traceability integrates
(1) the establishment of one or more relationships to
well-stated references or standards, and
(2) the documented ‘history’ of a product or a
system.
Trends in Analytical Chemistry, Vol. 23, No. 7, 2004 Trends

Table 1. Overview of European and international guiding institutions and regulatory bodies with their guidelines and standards on traceability,
measurement uncertainty (MU) and related topics

Body Full name Guidance on References

Eurachem A focus for analytical chemistry in Europe Traceability [2,3]

CITAC Cooperation on international traceability in analytical MU [13,14]
chemistry Reference materials [7]
Validation [15]
IUPAC International Union of Pure and Applied Chemistry MU [4,8,16,17]
ISO International Standardization Organisation
AOAC International Association of Official Analytical Chemists
FAO/WHO: Codex/CCMAS Food and Agricultural Organization/World Health MU [18–21]
Organisation: Codex Committee on Methods of Analysis
and Sampling

EAL European Cooperation for Accreditation MU [22]

ILAC International Laboratory Accreditation Cooperation MU [23]

These two parts of the basic meaning can be found
again when defining traceability as a property or a
characteristic of different analytical facets. The different
concepts of traceability are shown in Fig. 3.
The most obvious definition of traceability is a ‘prop-
erty of the result of a measurement or the value of a
standard whereby it can be related to stated references,
usually national or international standards, through an
unbroken chain of comparisons all having stated un-
certainties’. The different elements and the practical use
of this basic definition will be explained in Section 4.2
below.
Traceability of a result is related to traceability of a
method, which in turn is linked to traceability of stan-
dards and traceability of the equipment used in the
analytical procedure (Fig. 3). A method is called trace-
able when it produces results (with their uncertainties)
that are characterized by a defined traceability to well-
stated references [24]. Walsh [25] defines traceable
methods as ‘validated official or standard methods or
validated methods which contain uncertainty state-
ments and which are embedded into a quality system
and anchored to a common reference point’. Traceability
among standards is considered as the most relevant basis
for traceability of results [26], as shown in Fig. 3.
Traceability of equipment is defined as ‘the detailed,
timely, and customised recording of installation,
malfunctioning and repairs, periodic calibration and
corrections (if needed), hours of use, samples processed,
standard used, etc., in such a way that all questions
(what?, how?, who?, etc.) should have a detailed answer
in the pertinent documents’ [24].
Calibration is the set of operations used to establish
the relationship between values shown by a measuring
instrument and the values of measurement standards.
By calibrating, the results of measurements are related to
and thus made traceable to values of standards or
references. In practice, calibration is performed by
measuring samples with known amounts of analyte,
such as CRMs, and monitoring the measurement
response [2,3]. These definitions confirm the links between
traceability of equipment, standards and results (Fig. 3).

traceability of results
1. relationship(s) to
well-stated
references or
standards
traceability of methods extended meaning
of traceability
2. documented
'history' of a product
or a system
traceability of traceability of
standards equipment

Figure 3. Different concepts and extended meaning of traceability [24].

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Trends
4.2. Traceability in practice
The practical establishment of traceability is based on a
step-by-step implementation of the definition. The ISO
definition of traceability, originating from metrology (see
above), can be translated into three basic steps:
(1) to establish one or more links to well-stated refer-
ences,
(2) through an unbroken chain of comparisons, and
(3) to estimate all uncertainties associated with those
comparisons [12,24,27].
This definition is very much in line with the more
practical definition described by Eurachem/CITAC [3] in
its procedure for traceability that consists of the follow-
ing steps:
(1) specifying the measurand, the scope of measure-
ment and the required uncertainty;
(2) choosing the method of measurement;
(3) validating the method of measurement;
(4) identifying/quantifying all influences that will
affect the result;
(5) choosing appropriate references;
(6) estimating the uncertainty components associ-
ated with all influences and references [3].
The key principle in both approaches is that rela-
tionships to stated references are established and that
this is done through an unbroken chain of comparisons.
Practically, this means that the analytical procedure is
first described as a chain or a flow diagram (step (2) in
ISO definition; steps (1) and (2) in Eurachem/CITAC
definition). The word unbroken means that there is no
loss of information when considering the different steps
in the analytical procedure leading to the measurement
result. Each step in the procedure then needs to be linked
to either a reference method, a RM or an SI unit (step (1)
in-house/working RMs
higher level of traceability
certified RMs
primary RMs
SI units
Figure 4. The traceability chain and the relationship between traceability and uncertainty of measurements. The three possibilities for establishing
traceability referred to in Fig. 5, are indicated in bold [25,28].
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Trends in Analytical Chemistry, Vol. 23, No. 7, 2004
in ISO definition; step (5) in Eurachem/CITAC definition)
[12,24,27].
Fig. 4 depicts the successive classes of stated references
(materials or methods) in a so-called ‘traceability chain’.
Establishing traceability through a traceability chain
brings a certain level of uncertainty, called ‘calibration
uncertainty’ or ‘traceability uncertainty’ (see also
Section 6 below) [3,7,28].
This then brings us to the third key element when
applying definitions of traceability – the stated uncer-
tainties. Each step in the traceability chain, with the
uncertainty components of all the stated references, will
contribute to the measurement result and thus to the
uncertainty associated with it. Uncertainty components
must thus be estimated at each step in the analytical
process (step (3) in ISO definition; step (6) in Eurachem/
CITAC definition).
As described above, validation is a tool to identify all
possible effects or factors within the analytical procedure
that can influence the final result. As such, steps (3) and
(4) in the Eurachem/CITAC definition are additional
steps that can be very helpful in establishing traceability
[3].
Examples of how traceability is established in practice
can be found in the literature. Recently, a Special Issue
of TrAC was published on ‘Challenges for achieving
traceability of environmental measurements’ (TrAC,
Volume 23, 2004). This issue contains a lot of up-to-
date information and practical examples in the particu-
lar domain of environmental analysis. Many authors
reported on the most important and most difficult step in
establishing traceability – the selection of stated refer-
ences or standards. For different stages in the traceability
chain shown in Fig. 4, descriptions and examples are
given by Quevauviller and Donard [27], Charlet and
Marschal [29] and Segura et al. [30]. Pan [28], F€ orstner
[31] and Theocharopoulos et al. [32] applied the ISO
definition for establishing traceability in different types of
in-house/working method
higher level of method bias
spiking
and uncertainty
reference method
primary method
Trends in Analytical Chemistry, Vol. 23, No. 7, 2004 Trends

environmental methods of analysis. A similar approach error have been taken into account [37]. Within this
was followed by Sabe and Gauret [33] and Drolc et al. interval, the result is regarded as being accurate, i.e.,
[34]; however, they based it upon the Eurachem/CITAC precise and true [11].
Guide on Traceability [3]. In their examples, all the It cannot be overemphasized that MU is different from
authors took into account specific steps or influences in error. The error of an individual analytical result, the
the analytical procedure that can lead to a ‘broken’ difference between the result and the true value of
chain of comparisons, such as sampling and sample the measurand, is always a single value [38]. Part of the
treatment or preparation steps. Some authors reported value of a known error, the systematic error, can be used
on uncertainties associated in particular with sampling to correct a result. This means that, after correction, the
[35,36]. result of an analysis may be very close to the true value.
However, the uncertainty of the measurement may still
be very large, because there is doubt or limited knowl-
5. The concept of MU edge about how close the result is to the value. Uncer-
tainty is expressed as a range and applies to an
MU is the most important criterion in both method analytical procedure and a specific sample type, but to
validation and IQC. It is defined as ‘a parameter, asso- different determinations and thus measurement results.
ciated with the result of a measurement, that charac- The value of the uncertainty cannot be used to correct a
terizes the dispersion of the values that could reasonably measurement result.
be attributed to the measurand’ [11,14]. The measurand The error of an analytical result is related to the
refers to the particular quantity or the concentration of (in)accuracy of an analytical method and consists of a
the analyte being measured. The parameter can be a systematic component and a random component (Fig. 5)
standard deviation or the width of a confidence interval [14]. Precision and bias studies form the basis for eval-
[14,37]. This confidence interval represents the interval uation of the accuracy of an analytical method [18]. The
on the measurement scale within which the true value accuracy of results relates only to the fitness-for-purpose
lies with a specified probability, given that all sources of of an analytical system, assessed by method validation.

analytical result

true value difference between

error
analytical result and
true value
inaccuracy

expected value
(limiting mean)
systematic error random error
difference between difference between
expected value and bias analytical result and imprecision
true value expeted mean value

run effect reproducibility intermediate repeatability

persistent bias
variations within the whole analytical
system, over longer periods
precision
variations during a inter-laboratory within-lab variation due to inter-assay precision=
particular run variation, tested by random effects= variability variability over a short
collaborative studies over a longer period of time, time interval, under the
under different conditions same conditions

matrix variation method laboratory run random

effect bias bias bias bias

indicator for difference between TRUENESS indicator for difference between PRECISION
expected value and true value result and expeced value

analysis of CRMs + duplicate analysis

statistical control

single-laboratory validation

minimally needed in method validation

Figure 5. Composition of the error of an analytical result related to trueness and precision [4,8].

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However, reliability of results has to do with more than error components, forming together the ‘ladder of
method validation alone. errors’:
MU is more than just a single figure expression of
accuracy. It covers all sources of errors that are relevant (1) the method bias, a systematic error associated
for all analyte concentration levels. MU is a key indicator with the method as such;
of both fitness-for-purpose and reliability of results, (2) the laboratory bias, which is either a systematic
binding together the ideas of fitness-for-purpose, QC and error – if the laboratory is considered on its
thus covering the whole QA system [4,37]. own, or a random error – if the laboratory is
The MU of an analytical procedure is thus derived considered as one of a group, as is the case in in-
from, but differs from, the error of a single analytical terlaboratory studies;
result. The deviation of the measurement result from (3) the run error, seen as a systematic error for one
the true value comprises a number of systematic and run and as a random variation over several runs
random errors as shown in Fig. 5. Each of these error performed intralaboratory;
components adds its own uncertainty to the total (4) the repeatability error, which is a random error
uncertainty budget of the analytical procedure. The from the replicate measurements performed
different error components are therefore referred to as within a single run [10].
‘sources of uncertainty’. Depending on the sources of
uncertainty taken into account and thus the conditions As the error being considered applies for only a
of the measurement, the overall MU will be different and specified concentration of analyte isolated from a speci-
another definition of MU will apply. This means that fied type of sample or matrix, sampling errors and matrix
there is no single, straightforward definition of MU. It is variation effects are not included here [4].
rather a concept, the interpretation of which changes The more traditional distinction between error compo-
according to the measurement conditions and to the nents is between random errors and systematic errors
reference to which the result is traceable [10]. The dif- (Fig. 5). In this classical approach, random errors are
ferent definitions of MU are subjects of the following generally referred to as ‘precision’ (repeatability, inter-
section. mediate precision and reproducibility), while systematic
errors are typically attributed to the uncertainty on the
bias-estimate and calibration uncertainty. To this classi-
6. Different operational definitions of MU fication, other uncertainty contributions are added, such
as sampling effects, matrix effects and uncertainties
As illustrated in Fig. 6, the error of an analytical result associated with certain assumptions that underlie the
for a specified analyte concentration comprises different measurement method and/or the calculation equation [2].

Result = true value + method bias + lab bias + run error + repeatability error

Result = true value + traceability U + U on estimated bias + lab bias + run error + repeatability error
systematic error random error

random error
intermediate precision

Uncertainty = within-laboratory uncertainty

random error
reproducibility precision

Bias is estimated with: 1. reference method U (reference) = 0 difference between

reproducibility uncertainty
two methods
2. spiking U (spike) ~ 0 difference between
two measured results intermediate precision

bias-included uncertainty
3. certified reference U (CRM) = 0 difference between
material (CRM) measured & certified value intermediate precision

absolute uncertainty

Figure 6. Composition of the error of an analytical result related to measurement uncertainty. Different operational definitions of measurement
uncertainty [10]. Below on the left are the three possibilities for establishing traceability (see also Fig. 4).

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Trends in Analytical Chemistry, Vol. 23, No. 7, 2004
As mentioned above, each of these error components is
a potential source of uncertainty. Depending on the con-
ditions under which the analysis is performed, different
sources of uncertainty contribute to the overall uncer-
tainty. Hund et al. [10] introduced different operational
definitions of uncertainty, according to the number and
type of uncertainty sources considered (Fig. 6):
(1) within-laboratory uncertainty, derived from
intermediate precision and including only the
repeatability error and the run error;
(2) reproducibility uncertainty, derived from repro-
ducibility precision (interlaboratory tests) and
accounting for the repeatability error, run and
laboratory effects;
(3) bias-included uncertainty and absolute uncer-
tainty additionally take into account the method
bias, which is the most important source of uncer-
tainty because it refers to a reference or a stan-
dard, to which the method is considered to be
traceable. If the working method is not a primary
method – which is traceable to SI units (see Fig. 4)
– the method is always compared to another, ref-
erence method or is applied using appropriate
CRMs. This reference or standard needs to be con-
sidered when the uncertainty associated with the
method bias is estimated. In addition to the uncer-
tainty associated with this reference or standard,
there is the uncertainty on the estimated bias
(Fig. 6). The different possibilities of bias estima-
tion – and thus of traceability – are depicted in
Fig. 5. If the method is compared to a reference
method, the uncertainty associated with this ref-
erence method is considered negligible and the
bias is estimated as the difference between the
two methods (case 1 in Fig. 6). If there is no
method to compare with, bias can be estimated
by spiking samples and assessing the difference
between the spiked sample and the measured
sample. In this case also, the uncertainty on the
spike will approximate to zero (case 2) and the
only method bias is the difference between
the measured sample and the spiked sample. Ab-
solute uncertainty can be estimated only if CRMs
are used (case 3). Only in this case can full trace-
ability to SI units be guaranteed [10].
7. Approaches to establishing MU
In general, to estimate the overall uncertainty on a
particular result, it is necessary to know:
(1) all uncertainties arising from the measurement
procedure itself;
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(2) all uncertainties associated with the references or
standards, the analytical results are made trace-
able to [3].
Different approaches exist for the estimation of overall
MU, as reviewed by several authors [9,10,39] and
summarized in Table 2.
The most well-known, traditional approach is based
on identifying, quantifying and combining all individual
contributions to uncertainty. In this ‘bottom-up
approach’, the overall uncertainty is derived from the
uncertainties of the individual components. The com-
ponent-by-component assessment of MU was originally
developed for physical measurements and adopted by
Eurachem for chemical measurements [13]. However,
because of its complexity, this methodology has signifi-
cant costs in time and effort and has never found wide-
spread applications.
A simplified approach to assessing MU is the
‘fitness-for-purpose approach’, defining a single
parameter called the fitness function. This fitness
function has the form of an algebraic expression
u ¼ f ðcÞ and describes the relationship between the
MU and the concentration of the analyte. For example
u ¼ 0:05c means that the MU is 5% of the concen-
tration. Calculation of the MU will hereby rely on
data obtained by evaluating individual method-
performance characteristics, mainly repeatability and
reproducibility precision, and preferably also bias
[21,40,41]. This approach can more or less be seen as
a simplification of the step-by-step protocol for testing
the MU, as described by Eurachem [14].
Although MU comprises more than systematic and
random errors, it can be estimated from method-valida-
tion data. Data from method-performance studies can
give all or nearly all the information required to evaluate
the uncertainty [2,4,18,37]. This includes the use of
data from in-house and collaborative validation studies
(typically precision data), proficiency-testing schemes
(typically bias data) or QA data, relevant for uncertainty.
If such data are available and used to estimate the un-
certainty, it is not necessary to estimate MU using the
component-by-component approach [18,19]. In partic-
ular, validation studies and QC measures are considered
highly relevant sources for estimating MU [42,43]. Table
2 describes three methodologies for assessing MU based
on validation data.
In the Analytical Methods Committee’s ‘top-down
approach’ [37], the laboratory is seen from a higher
level, as a member of a population of groups. As a
consequence, systematic errors within one laboratory
become random errors and the estimated uncertainty is
the reproducibility uncertainty (Fig. 6). Examples of
MU-estimation studies using data from collaborative ring
trials are works by Dehouck et al. [44] and Maroto et al.
[45,46].
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Trends Trends in Analytical Chemistry, Vol. 23, No. 7, 2004

Table 2. Different approaches for estimating measurement uncertainty (MU)

Reference Name of approach Basic principle Strengths Weaknesses

Eurachem [13] Bottom-up, error-budget, Identification, quantification and Holistic ¼ all important Complex, Expensive,
and ISO [16] error-propagation or combination of all sources of sources of error should be Time-consuming
component-by- uncertainty included
component

Codex Fitness-for-purpose Establishment of a fitness-function, Simple MU can be assessed Some sources of

Alimentarius/ u ¼ f(c), based mainly on precision for different concentrations uncertainty may be
CCMAS [18–21] and bias studies overlooked
Analytical Top-down Based on data obtained from MU can be assessed for Some sources of
Methods inter-laboratory studies (precision) different concentrations uncertainty may be
Committee [37] overlooked
Only if data on
collaborative studies
are available
Eurachem [14] Validation-based Based on inter- or intra-laboratory Extension of validation work, Some sources of
Barwick & validation studies (precision, so no extra work is needed uncertainty may be
Ellison [47] trueness, robustness) overlooked
Hund et al. [39] Robustness-based Based on robustness tests as Simple, time-efficient Some sources of
intra-laboratory simulations of uncertainty may be
inter-laboratory studies overlooked
Method must first show to
be robust

The other two approaches mentioned in Table 2
[14,39,47] make use of different method-performance
parameters. All three validation-based methodologies
can be seen as simpler, and more time- and cost-efficient
extensions of validation.
However, it is important to note that not all sources of
uncertainty are covered by method-performance data.
Some sources that may need particular consideration in
addition to the available data are sampling, pre-treat-
ment, method bias, variation in conditions and changes
in the sample matrix [14,18,41].
Many of those principles for estimating MU were
applied in a case study for toluene in ground water
performed by Armishaw [48]. His idea was that, for a
routine method that has been validated previously and
performed in a laboratory where QC measures are in
place, it is possible to estimate MU in a working after-
noon. The key is to extract all relevant information from
already available data: validation studies (bias/recovery
and precision data); instrument calibration data (RM
uncertainties); and, information from regularly per-
formed QC measurements (replicate analyses and control
samples) and from the method procedure itself
(sampling, homogeneity of samples,. . .). After identifying
the components contributing to the uncertainty budget
and assigning the relevant sources of information,
standard uncertainties were quantified and combined to
form the combined standard uncertainty.
The results for toluene in water were reported as
x
U, where U was the expanded uncertainty obtained
by multiplying the combined standard uncertainty, uc ,
with a coverage factor of 2. In addition to this bottom-up
MU estimation (Approach 1), Armishaw reported the
expanded uncertainty as a function of the concentration
of toluene (Approach 2). Finally, the authors compared
this experimentally assessed MU with calculated MU
values, based on: (1) a within-laboratory reproducibility
estimate; (2) proficiency test data; (3) the models of
Horwitz [49] and Thompson and Lowthian [50]. These
models allowed calculation of % RSD values as a func-
tion of the analyte concentration. To obtain expanded
uncertainty values, Armishaw multiplied the predicted
SD values from the models by 2 [48]. All three MU
calculations are variants of validation-based approaches
(Approaches 3 and 4, Table 2).
8. Importance of traceability and MU
The underlying motivation to establish traceability and
MU is the need to make decisions based on the analytical
results obtained or to be in compliance with regulatory
limits (for quantitative determinations) [51]. MU is an
essential feature of analytical results, for three different
reasons:
(1) Customers want to have an idea about the range of
results and about the comparability of results
between different laboratories [43]. Any result
must be accompanied by a statement of MU, so that
the user of the result knows the level of confidence
associated with it [52]. The concepts of compara-

488 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 23, No. 7, 2004
bility and reliability of results have been discussed
briefly in Section 1 and are presented in Fig. 1.
(2) It demonstrates traceability. Before MU can be eval-
uated, traceability to stated references or standards
must be established. Moser et al. [43] claim that
traceability is proved by the appropriate use of
RMs or standards and by a full uncertainty budget.
(3) There is a requirement to know the method, to
understand the underlying principles and
mechanisms of the measurement procedure. Lack
of profound knowledge on the method itself
will lead to certain unknown uncertainty
contributions not being taken into account and
thus to gaps in the uncertainty budget. MU can
be estimated only if the method is well understood
[43,53].
MU is increasingly gaining attention, in particular
within the framework of accreditation. The new
accreditation standard ISO/IEC 17025 [17], which has
been in force from December 2002, contains clear
requirements about estimating MU and when and how it
should be stated in test reports. ISO/IEC 17025 requires
MU to be reported when required by the client and when
relevant to the application and the interpretation of the
measurement results, within the framework of certain
specifications or decision limits. The MU should be readily
available and reported together with the result as X
U,
where U is the expanded uncertainty [17,47,51,54].
Eurachem and CCMAS within the Codex Alimentarius
deal with MU as a separate issue [14,18–20].
The Analytical Methods Committee even claims that
MU will become the main unifying principle of analytical
data quality [37].
9. Summary
Rather than focusing on the techniques and methodol-
ogies being used, attention is nowadays paid to the
quality and the reliability of the final results of analysis.
This is influenced by greater demand for regulatory
compliance and greater awareness of the customer – the
client wants to know the level of confidence of the
reported result. In order for results to be comparable,
they must be reported with a statement of MU and they
must be traceable to common primary references.
Methods must be validated to show that they actually
measure what they are intended to measure; that they
are fit for exacting European and international stan-
dards, such as the ISO/IEC 17025 norm for laboratory
accreditation. On the basis of quality and reliability of
analytical data rests the comparability of results for their
specific purpose.
An analytical method is a complex, multi-step process,
starting with sampling and ending with the generation
Trends
of a result. Although every method has its specific scope,
application and analytical requirement, the basic prin-
ciples of QA are the same, regardless the type of method
or the sector of application. The information in this
article is taken mainly from the analytical chemistry, but
it also applies to other sectors. The validation of
analytical methods, the establishment of traceability of
results and the assessment of MU should be done in a
uniform, harmonized way, conforming with interna-
tionally recognized standards from institutions such as
Eurachem, IUPAC or ISO. This update on analytical
quality provides a common understanding for the topics
of method validation, traceability and MU of measure-
ments. It has elucidated the interrelationships between
method validation and traceability and MU of results.
From all the guidelines and standards, we selected and
summarized the most relevant information. We dis-
cussed different approaches to establishing traceability
and assessing MU of analytical methods in general. We
highlighted the importance of both concepts and the link
with method validation and analytical QA.
Acknowledgements
We wish to thank Andrew Damant for giving sugges-
tions and Friedle Vanhee for reading and assistance.
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Isabel Taverniers graduated in Agricultural and Applied Biological
Sciences from the University of Gent, Belgium, in 1999. Until April
2001, she worked at AgriFing, a joint spin-off laboratory of Gent
University, Hogeschool Gent and the Department for Plant Genetics and
Breeding (DvP), where she specialized in DNA fingerprinting technol-
ogies. She is now preparing a Ph.D. thesis in the Laboratory of Applied
Plant Biotechnology of the Department for Plant Genetics and Breeding
(CLO, Flemish Community).
Erik Van Bockstaele is Head of the Department for Plant Genetics
and Breeding and Professor at the Faculty of Agricultural and Applied
Biological Sciences of the University of Gent.
Marc De Loose is Head of the Section Applied Plant Biotechnology at
the Department for Plant Genetics and Breeding.