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.
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, Systeme 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
doi:10.1016/S0165-9936(04)00733-2
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
MU estimation 2. Which of the 2 results
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
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], Forstner€
[31] and Theocharopoulos et al. [32] applied the ISO
definition for establishing traceability in different types of
in-house/working method
bias
spiking
reference method
primary method
 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 Trends

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

analytical result
difference between
true value error

analytical result and

true value
inaccuracy

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

persistent bias run effect reproducibility intermediate repeatability

precision
variations within the whole analytical variations during a inter-laboratory within-lab variation due to inter-assay precision=

system, over longer periods particular run variation, tested by random effects= variability variability over a short
collaborative studiesover 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 indicator for difference between

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

bias-included uncertainty
3. certified referenceU (CRM) = 0 difference between

material (CRM)
measured & certified valueintermediate 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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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.