EURACHEM / CITAC Guide

Measurement uncertainty
arising from sampling
A guide to methods and approaches

Produced jointly with

EUROLAB, Nordtest and the
UK RSC Analytical Methods Committee

First Edition 2007

 Measurement uncertainty arising from sampling:
A guide to methods and approaches

Produced jointly by
Eurachem, EUROLAB, CITAC, Nordtest and the RSC Analytical Methods
Committee

Editors
Michael H Ramsey (University of Sussex, UK)
Stephen L R Ellison (LGC, UK)

Composition of the Working Group

Eurachem members
Michael H Ramsey (Chair) University of Sussex, UK
Stephen L R Ellison (Secretary) LGC, UK
Paolo de Zorzi National Environmental Protection Agency, Italy
Pentti Minkkinen Lappeenranta University of Technology, Finland
Roger Wood Food Standards Agency, UK
Mire C Walsh State Chemist for Ireland
Alex Williams UK
Christian Backman GTK, Finland
Maria Belli National Environmental Protection Agency, Italy
EUROLAB members
Manfred Golze BAM, Germany
Rdiger Kaus HS Niederhein, Denmark
Ulrich Kurfrst FH Fulda, Denmark
Mikael Krysell Eurofins Denmark
CITAC member
Ilya Kuselman National Physical Laboratory, Israel
Nordtest representatives
Christian Grn DHI Water & Environment, Denmark
Acknowledgements Bertil Magnusson Technical Research Institute of Sweden
Astrid Nordbotten Food Safety Authority, Norway
This document has been produced primarily by a
joint Eurachem/EUROLAB/CITAC/Nordtest/
AMC representatives
Michael Thompson University of London, UK
AMC Working Group with the composition
Jenny Lyn Food Standards Agency, UK
shown (right) working in the period 20042006.
The editors are grateful to all these individuals Observer
and organisations and to others who have Marc Salit NIST, USA
contributed comments, advice and assistance.
Production of this Guide was in part supported This publication should be cited as:
under contract with the UK Department of Trade
M H Ramsey and S L R Ellison (eds.) Eurachem/EUROLAB/
and Industry as part of the National Measurement
System Analytical Measurement (VAM) CITAC/Nordtest/AMC Guide: Measurement uncertainty arising from
Programme, and also by the UK Food Standards sampling: a guide to methods and approaches Eurachem (2007). ISBN
Agency and Environment Agency. 978 0 948926 26 6. Available from the Eurachem secretariat

UfS:2007 Page i
Foreword
Uncertainty of measurement is the most important single parameter that describes the
quality of measurements. This is because uncertainty fundamentally affects the
decisions that are based upon the measurement result. Substantial progress has been
made in devising procedures to estimate the uncertainty that originates in the
analytical portion of the measurement, and guidance on these procedures is available
[1]. However, a measurement almost invariably involves the process of taking a
sample. This is because it is usually impossible to analyse the entire bulk of the
material to be characterised (the sampling target). If the objective of the
measurement is to estimate the value of the analyte concentration in a sampling target,
then the uncertainty associated with the sampling process must inevitably contribute
to the uncertainty associated with the reported result. It has become increasingly
apparent that sampling is often the more important contribution to uncertainty and
requires equally careful management and control. The uncertainty arising from the
sampling process must therefore be evaluated. While existing guidance identifies
sampling as a possible contribution to the uncertainty in a result, procedures for
estimating the resulting uncertainty are not well developed and further, specific,
guidance is required.
Historically, measurement scientists have been primarily concerned with
measurements made within laboratories, and the process of sampling has been
conducted by, and the responsibility of, a different set of people who are often in
separate organisations. The measurement scientists knowledge of the sampling
process is then very limited. Conversely, the advent of in situ analytical techniques
sometimes enables the measurement scientist to make measurements at the sampling
site and in contact with the material to be sampled. Examples of this situation are
process analysis within industrial production, and in situ measurements on
contaminated land. The placing of the analytical sensor in these situations then
constitutes the taking of a sample, and the measurement scientist becomes not only
aware of, but responsible for, all stages of the measurement process, including the
sampling. Such an awareness of the whole process is important, irrespective of the
division of effort. Since analytical and sampling processes contribute to the
uncertainty in the result, the uncertainty can only be estimated if there is an
understanding of the complete process. Further, optimisation of the relative effort in
sampling and analysis is only possible where sampling and analytical processes are
both understood.
If the different stages are the responsibility of different people, there needs to be good
communication between all of the parties involved. Sampling planners and analytical
scientists need to optimise the whole measurement procedure, and to devise a strategy
to estimate the uncertainty. Both need to discuss the objectives of the measurements
with the customer. All three parties need guidance from the appropriate regulator on
how these estimates of uncertainty are to be acted upon, to ensure the reliability of the
decisions based upon the measurements. To underpin these decisions, all the parties
need reliable estimates of uncertainty, including that arising from sampling. Although
no general guidance can replace expert advice in complex or critical cases, this Guide
describes the methods needed to fulfil the need for reliable estimates of uncertainty
from sampling for most analytical measurement systems.

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Summary
This Guide aims to describe various methods that can be used to estimate the
uncertainty of measurement, particularly that arising from the processes of sampling
and the physical preparation of samples. It takes a holistic view of the measurement
process to include all of these steps as well as the analytical process, in the case where
the measurand is defined in term of the value of the analyte concentration in the
sampling target, rather than in just the sample delivered to the laboratory. The Guide
begins by explaining the importance of knowing the total uncertainty in a
measurement for making reliable interpretation of measurements, and judging their
fitness for purpose. It covers the whole measurement process, defining each of the
component steps, and describing the effects and errors that cause uncertainty in the
final measurement.
Two main approaches to the estimation of uncertainty from sampling are described.
The empirical approach uses repeated sampling and analysis, under various
conditions, to quantify the effects caused by factors such as the heterogeneity of the
analyte in the sampling target and variations in the application of one or more
sampling protocols, to quantify uncertainty (and usually some of its component parts).
The modelling approach uses a predefined model that identifies each of the
component parts of the uncertainty, making estimates of each component, and sums
them in order to make an overall estimate. Models from sampling theory can
sometimes be used in this approach to estimate some of the components from a
knowledge of the characteristics of particulate constituents.
Worked examples are given of each of these approaches, across a range of different
application areas. These include investigations of the environment (of soil and water),
of food (at growing and processing) and of animal feed. The estimates of the total
uncertainty of measurement range from a few per cent up to 84% relative to the
measurand. The contribution of the sampling is occasionally small but is often
dominant (>90% of the total measurement variance). This suggests an increased
proportion of the expenditure needs to be aimed at the sampling, rather than the
chemical analysis, if the total uncertainty needs to be reduced in order to achieve
fitness for purpose.
Management issues addressed include the responsibility of the quality of the whole
measurement process, which needs to include the sampling procedure. Guidance is
given on the selection of the most appropriate approach for any application, and
whether one initial validation of the system is sufficient, or whether there is a need for
ongoing monitoring of the uncertainty from sampling using quality control of
sampling. The extra cost of estimating uncertainty is also considered in relation to the
cost savings that can be made by knowing the uncertainty of measurement more
reliably.
Such a Guide can never be fully comprehensive, and although there are appendices
with details of some of the statistical techniques employed and sources of more
detailed advice, there will often be a need for expert advice in more complex
situations. This Guide aims to be a useful introduction to this subject, but we hope it
will also stimulate further research into improved methods of uncertainty estimation.

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Contents

Foreword ii

Summary iii

List of acronyms and abbreviations vii

PART 1 Introduction and scope 1

1 Introduction 1
1.1 Rationale for the Guide 1
1.2 Aim of the Guide 1
1.3 Application to judging fitness for purpose 1
1.4 Intended audience for the Guide 2
1.5 Relationship of this Guide to other documents 2
1.6 Using the Guide 3

2 Scope and field of application 3

3 Terminology 4

PART 2 Fundamental concepts 5

4 Sampling in the measurement process 5

5 Uncertainty of measurement 6
5.1 Definition of uncertainty of measurement 6
5.2 Specification of measurand 8
5.3 Error, precision and uncertainty 9
5.4 Sampling and physical preparation as sources of uncertainty of measurement 9
5.5 Sources of uncertainty 10
5.6 Heterogeneity as a source of uncertainty 10

PART 3 Estimation of measurement uncertainty including sampling 13

6 Approaches to uncertainty estimation 13

7 The measurand 14

8 General conduct of studies of sampling uncertainty 14

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9 Empirical approach 14
9.1 Overview 14
9.2 Uncertainty sources 14
9.3 Statistical model for the empirical estimation of uncertainty 15
9.4 Empirical estimation of uncertainty 16
9.5 Calculation of uncertainty and its components 18
9.6 Alternative empirical methods of uncertainty estimation 19

10 The modelling approach 20

10.1 Cause-and-effect modelling 20
10.2 Sampling theory for estimation of uncertainty 20

PART 4 Management issues 24

11 Responsibility for quality of sampling 24

12 Selection of uncertainty estimation approach 24

13 Quality control of sampling 25

13.1 Relationship between validation and quality control 25
13.2 Methods of internal quality control of sampling 26

14 Reporting and interpreting uncertainty 27

14.1 Introduction 27
14.2 Expanded uncertainty, U 27
14.3 Relative uncertainty statements 28
14.4 Contributions to uncertainty 28
14.5 Applicability of estimates 29
14.6 Interpretation of uncertainty statements against limits 29

15 Cost of estimating uncertainty from sampling 29

16 Judging fitness for purpose of measurements using uncertainty 30

17 Implications for planning sampling and measurement strategies 32

17.1 Expertise and consultation 32
17.2 Avoiding sampling bias 32
17.3 Planning for uncertainty estimation 32
17.4 Fitness-for-purpose criteria 32
17.5 Use of prior validation data 33
17.6 Acceptability of sampling uncertainty 33

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Appendix A: Examples 34
Example A1: Nitrate in glasshouse grown lettuce 35
Example A2: Lead in contaminated top soil 40
Example A3: Dissolved iron in groundwater 46
Example A4: Vitamin A in baby porridge containing fruit and milled cereals 55
Example A6: Cadmium and phosphorous in agricultural top soil by modelling approach
74

Appendix B: Terminology 83

Appendix C: Useful statistical procedures 91

C1. Estimating bias between two sampling methods, by using paired samples 91

C2. Further description of sampling errors from sampling theory 93

C3. Sources of software for calculations 96

Appendix D: Alternative experimental designs for empirical uncertainty

estimation 97

1. Multi-level designs to estimate other component effects 97

2. Simplified and unbalanced designs, to reduce the cost of implementation 98

Appendix E: Modifying sampling uncertainty using predictions from sampling

theory 99

Appendix F: References 100

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List of acronyms and abbreviations

ANOVA analysis of variance

AQC analytical quality control
BIPM Bureau International des Poids et Mesures
CEN European Committee for Standardization
CH constitution heterogeneity
CRM certified reference material
CTS collaborative trial in sampling
df degrees of freedom
DH distribution heterogeneity
FAPAS trade name of body that organises international proficiency tests
FSE fundamental sampling error
GEE global estimation error
GFAAS graphite furnace atomic absorption spectrometry
GSE grouping and segregation error
GUM Guide to the Expression of Uncertainty in Measurement. ISO
HPLC high performance liquid chromatography
IDE increment delimitation error
IEC International Electrotechnical Commission
IFCC International Federation of Clinical Chemistry and Laboratory Medicine
IPE increment and sample preparation error
ISO International Organization for Standardization
IUPAC International Union of Pure and Applied Chemistry
IUPAP International Union of Pure and Applied Physics
IXE increment extraction error
LOD limit of detection
LOQ limit of quantification
MU measurement uncertainty
NIFES National Institute of Nutrition and Seafood Research
NIST National Institute of Standards and Technology
OIML International Organization of Legal Metrology
PSE point selection error
PME point mineralisation error
PT proficiency test
QA quality assurance
QC quality control
RANOVA robust analysis of variance
RSD relative standard deviation
RST reference sampling target
SD standard deviation
SPT sampling proficiency test
SS sum of squares
SWE weighting error
TAE total analytical error
TSE total sampling error

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 Introduction

PART 1 Introduction and scope

1 Introduction
1.1 Rationale for the Guide
The main purpose of measurement is to enable decisions to be made. The reliability of these
decisions depends on knowing the uncertainty of the measurement results. If the uncertainty
of measurements is underestimated, for example because the sampling is not taken into
account, then erroneous decisions may be made that can have large financial consequences.
The fitness for purpose of measurement results can only be judged by having reliable
estimates of their uncertainty. For this reason it is essential that effective procedures are
available for estimating the uncertainties arising from all parts of the measurement process.
These must include uncertainties arising from any relevant sampling and physical preparation.
Judgements on whether the analytical contribution to the uncertainty is acceptable can only be
made with knowledge of the uncertainty originating in the rest of the measurement procedure.

1.2 Aim of the Guide

1.2.1 The aim of this Guide is to explain the rationale, and practical application, of the
methods available for the estimation of uncertainty that includes the contribution from
sampling. The Guide does not aim to recommend individual sampling protocols, which are
often prescribed in other documents or regulations, but rather to consider the measurement
uncertainty generated by whatever protocol is employed.
1.2.2 The Guide also aims to explain the importance of sampling to the overall uncertainty
budget, and hence to the reliability of the consequent decisions made using the measurements.
As well as explaining how to estimate the uncertainty, the Guide will explain the justification
for including sampling in the overall management of the measurement process.
1.2.3 Unlike the assumption that is often made for estimates of uncertainty for an
analytical method, an estimate for one sampling protocol for one batch of material should not
be assumed as automatically applicable to any subsequent batch of material. For example,
depending on the sampling target, the degree of heterogeneity (i.e. inhomogeneity) may have
changed substantially. There will be a need, therefore, for routine monitoring of key
parameters of sampling quality to examine and update estimates of uncertainty for subsequent
batches.

1.3 Application to judging fitness for purpose

One of the main benefits of knowing the uncertainty of a measurement is to enable a
stakeholder to judge its fitness for any particular purpose. A proper understanding of
uncertainty from sampling must therefore be embedded in the broader perspective of fitness
for purpose. This is important for two reasons. Firstly, it ensures that the estimate of
uncertainty of each measurement is realistic when compared with the optimal value of
uncertainty required to give reliable decisions. Secondly, given this level of uncertainty that is
required to be fit for purpose, it is necessary to distribute effort (or expenditure) between the
sampling and the analytical aspects of the measurement process in order to obtain the required
uncertainty most economically. These ideas are developed further, and a quantitative
approach to judging fitness for purpose by balancing uncertainty against cost is introduced, in
Section 16.

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 Introduction

1.4 Intended audience for the Guide

This Guide is intended primarily for specialists such as sampling planners and for analytical
chemists who need to estimate the uncertainty associated with their measurement results.
Other stakeholders should seek specialist advice for particular applications.

1.5 Relationship of this Guide to other documents

1.5.1 Current practice in the estimation of uncertainty for a broad range of measurements
follows the Guide to the Expression of Uncertainty in Measurement (the GUM) [2],
published in 1993 by ISO in collaboration with BIPM, IEC, IFCC, IUPAC, IUPAP and
OIML. The GUM sets out the concepts required, established the general principles, and
provided a procedure applicable to cases where an adequate model of the measurement
process is available. The application of this approach to chemical analysis was described in
1995 in a Eurachem Guide for Quantifying Uncertainty in Analytical Measurement [3], and
broadened to include the use of validation and method performance data in a second edition in
2000 [1]. Other useful contributions to the practical estimation of uncertainty of analytical
measurements using collaborative study data have been made by the Analytical Methods
Committee of the Royal Society of Chemistry in 1995 [4], and by ISO TC/69 in 2004 [5].
This Guide on sampling is consistent with the general principles established in the GUM.
1.5.2 Sampling theory has developed largely independently of analytical chemistry and
chemical metrology. Sampling quality has generally been addressed in sampling theory by the
selection of a correct sampling protocol, appropriate validation, and training of sampling
personnel (i.e. samplers) to ensure that this protocol is applied correctly [6]. It is then
assumed that the samples will be representative and unbiased, and the variance will be that
predicted by the model.
1.5.3 An alternative approach is to estimate the uncertainty of sampling for typical
materials, or for sampling targets, during validation of the sampling protocol, and to confirm
compliance in practice using ongoing quality control. This is more consistent with procedures
already in place for the rest of the measurement process. Interestingly, the quality of sampling
is only quantifiable through the measurements that are made upon the resultant samples.
1.5.4 Sampling protocols have been written to describe the recommended procedure for
the sampling of innumerable types of material and for many different chemical components.
These protocols are sometimes specified in regulation or in international agreementsa[7].
These procedures rarely identify the relative contributions of sampling and chemical analysis
to the combined uncertainty.b
1.5.5 There is accordingly a range of prior literature on the theory and practice of
sampling. As explained in Section 1.2.1, this Guide therefore does not seek to propose further
sampling protocols but rather to provide methodologies to quantify the uncertainty that arises
when a given protocol is used.
1.5.6 A handbook describing procedures for the estimation of uncertainty from sampling,
derived from this Guide but with further case studies, has been prepared by the Nordtest
group [8].

a
The acceptance sampling procedures are applied to the sampling of a wide range of materials [7].
b
Some concepts from sampling theory can be usefully adapted for estimation of uncertainty of measurement
(Section 10.2).

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 Scope and field of application

1.6 Using the Guide

1.6.1 This document summarises the concepts necessary for understanding uncertainty in
sampling, and provides procedures that allow their practical implementation. The Guide
additionally covers issues related to management, quality assurance and reporting of results
with uncertainty. The scope and intended field of application are set out in Section 2, which
also summarises the approaches covered. Terminology is discussed in Section 3, and key
terms defined in Appendix B.
1.6.2 Fundamental concepts are covered in Sections 4 and 5. An overview of the
measurement process is provided in Section 4. This includes an explanation of the sampling
terminology used, and indicates which steps in the process are considered in detail in this
Guide. Measurement uncertainty and its sources are discussed further in Section 5.
1.6.3 Sections 6 to 10 describe methodologies for the estimation of uncertainty, with a
discussion of the merits of the various options. The two broad approaches available are
summarised in Section 6, and covered in detail in Sections 1 and 10 respectively. The intent is
to provide a range of options that may be applied, rather than to specify any particular
approach.
1.6.4 Management and quality issues are addressed in Sections 11 to 1. These include a
very brief discussion of responsibilities for quality in sampling (Section 11) before discussing
the selection of uncertainty estimation approach in Section 1. The use of sampling quality
control to monitor sampling performance is covered in Section 1. Reporting and use of
uncertainty, and its effect on the reliability of decisions, are discussed in Section 14. Cost is
an important factor and selection of the most cost-effective and appropriate method of
estimation is explained in Section 15. Knowing the value of the uncertainty helps to judge the
fitness for purpose of the measurement as a whole, and its component parts, and this is
discussed in Section 16.
1.6.5 A range of examples, a detailed glossary of terms and definitions used in this Guide,
some important statistical procedures and experimental designs, and a discussion of
improving sampling uncertainty using predictions from sampling theory are provided as
appendices.

2 Scope and field of application

2.1 The principles of this Guide are applicable to the estimation of uncertainty from the
full range of materials that are subject to analytical measurement (e.g. gaseous, liquid and
solid). These include environmental media (e.g. rock, soil, water, air, waste and biota), foods,
industrial materials (e.g. raw materials, process intermediaries and products), forensic
materials and pharmaceuticals. This approach is applicable to sampling by any protocol,
whether it uses single or composite samples, or single or multiple determinations.
2.2 The Guide describes the estimation of uncertainty using i) replicated measurement
and sampling (the empirical approach) and ii) modelling based on identified influence
quantities and theoretical considerations (the modelling approach).
2.3 The use of uncertainty estimates in the assessment of fitness for purpose and in the
optimisation of effort among individual parts of the measurement process is covered. Methods
of assessing fitness for purpose that are described include those based upon percentage of
total variance and others based on cost-benefit analysis.

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 Terminology

2.4 Although the general principles of this Guide apply, it does not specifically discuss
microbiological sampling. Nor does it discuss the estimation of uncertainty in spatial or
temporal information such as the location or size of areas of high analyte concentration.

3 Terminology
3.1 The precise definitions of many of the terms used in this Guide vary depending on
the area of application. A full listing of terms and their different definitions is given in
Appendix B. In this Guide, normative definitions of each term have been selected that are as
generally applicable as possible to all sectors of application. These terms are listed in
Appendix B and in bold on first use in the text.

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 Sampling in the measurement process

PART 2 Fundamental concepts

4 Sampling in the measurement process

Figure 1: Schematic diagram of the typical measurement process

Process step Form of Description of process step

material

Sampling Sampling Target Collection of a single sample, or several

increments combined into composite sample

Primary Sample Comminution and/or splitting

Sub-sample Further comminution and/or splitting

Physical sample
preparation

Laboratory Physical preparation, e.g. drying, sieving,

sample milling, splitting, homogenisation

Test sample Selection of test portion for chemical

treatment preceding chemical analysis

Test portion Chemical treatment leading to analytical

determination
Analysis

Test solution Determination of analyte concentration

The figure shows a complete measurement process, starting with primary sampling and ending in the analytical
determination. There are many intermediary steps, such as transportation and preservation of samples, not all of
which are always present. Each step gives rise to a contribution towards the uncertainty of measurement. This
Guide concentrates on the process steps of sampling and physical sample preparation (shaded boxes), as the
last step is well covered in previous guidance [1]. Notice that two of the sampling steps occur within the
laboratory (light grey) and are frequently considered to be part of the analytical process. For definitions of terms
see Appendix B.

A sampling target is the portion of material, at a particular time, that the sample (and therefore
the measurement result) is intended to represent. The sampling target needs to be defined
prior to the design of the sampling plan. It may be defined by regulation, such as the whole of
a batch, lot or consignment. If the properties and characteristics (e.g. analyte concentration) of
the material in a certain area, or time period, are of interest and must be known, then it can be
considered a sampling target. When the composition of a whole batch is required (e.g. of a
food material), then the whole batch constitutes the target. When the spatial (or temporal)

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 Uncertainty of measurement

variation of concentration is required (e.g. in finding hot spots within a contaminated

material), then each location where the concentration is required will be a separate sampling
target. Any one sampling target will ultimately generate one reported measurement result and
an uncertainty.
Primary samples are often made up of a number of increments, which are combined to form
a composite sample before a measurement is made. It is the uncertainty on this single
measurement value, made on this composite sample, and caused by all of the preparatory
steps, that is required. The value of this uncertainty will often be affected by the number of
increments that are taken. This contrasts with the situation when several distinct primary
samples (n) are taken from different parts of the sampling target, and measured separately. If
the composition of the sampling target is calculated by taking the mean value of these
separate measurements, then the uncertainty on the mean value is calculated using the
standard error of the mean (s/n). This is not the same as the uncertainty on the single
measurement, the estimation of which is the objective of this Guide.
4.1 The whole process of measurement (Figure 1) typically begins with the taking of the
primary sample from a sampling target. The resulting sample goes through one or more of a
series of steps prior to the analytical determination. All steps contribute to uncertainty in the
final result, when the analyte value required (i.e. measurand value or true value), is
expressed in terms of the analyte concentration in the sampling target. Guidance already
exists on the estimation of the analytical steps of the measurement process [1]. This will
certainly include the selection of the test portion, the chemical treatment preceding
measurement and the analytical determination, but may also include the physical preparation
of the laboratory sample by means such as drying, sieving, milling, splitting and
homogenisation.
4.2 In common practice, all the various portions of material in the second column of
Figure 1 are often referred to simply as a sample. It is clearly important to differentiate them
carefully in discussion, especially those considered particularly in this Guide (in shaded boxes
on Figure 1). This is discussed in more detail in Section 5.2.
4.3 Methods described in the Guide will help to identify the dominant source of the
uncertainty, such as the sampling rather than the chemical analysis, but will not necessarily
explain the cause. However, heterogeneity within the sampling target, either spatial or
temporal, is known to be a significant cause of uncertainty in many circumstances. Separate
studies would be needed to characterise the variability that contributes to the uncertainty. For
the purpose of this Guide, heterogeneity within the sampling target is treated as just one cause
of uncertainty in the final measurement. This is the case, whatever actions are taken to
minimise the effects of the heterogeneity by the application of any particular sampling
protocol.

5 Uncertainty of measurement
5.1 Definition of uncertainty of measurement
5.1.1 Uncertainty of measurement, or measurement uncertainty (MU), is defined in
metrological terminology [2] as:

Parameter, associated with the result of a measurement, that characterises the dispersion of the
values that could reasonably be attributed to the measurand.

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 Uncertainty of measurement

The definition includes several important features, which are discussed in the following
paragraphs.
5.1.2 The parameter may be, for example, a range, a standard deviation, an interval (like
a confidence interval) or half-interval (u is a statement of a half-interval) or other measure of
dispersion such as a relative standard deviation. Note that when MU is expressed as a
standard deviation, the parameter is known as standard uncertainty, usually given the
symbol u. Other forms of expression are considered in Section 14.
5.1.3 Uncertainty is associated with each measurement result. A complete measurement
result typically includes an indication of the uncertainty in the form xU, where x is the
measurement result and U an indication of the uncertainty (it will be seen that the symbol U
has a special meaning, in this Guide; it indicates an expanded uncertainty, which will be
discussed further in Section 14). This form of expressing a result is an indication to the end-
user of the result that, with reasonable confidence, the result implies that the value of the
measurand is within this interval.
5.1.4 The measurand is simply a quantity, such as a length, mass, or concentration of a
material, which is being measured. The term value of the measurand is closely related to the
traditional concept of true value in classical statistical terminology. From this alternative
viewpoint uncertainty has also been defined [9] as:

An estimate attached to a test result which characterises the range of values within which the true
value is asserted to lie
This definition (which will be referred to as the statistical definition) has the advantage of
being easier to explain to decision makers, who often recognise the phrase true value as the
value of interest for their decision. It has the disadvantage that the true value itself can never
be known and this generally requires further explanation.
5.1.5 The metrological definition asserts that uncertainty expresses the dispersion of the
values that could reasonably be attributed to the measurand. This is a particularly important
phrase. It indicates that although the uncertainty is associated with a measurement result, the
range quoted must relate to the possible range of values for the measurand. For example, the
measurand could be the total mass of gold in a geological deposit. This is quite different from
a statement of precision, which would describe the range of results that might be observed if
the measurement were repeated. In requesting information about where the measurand value
might be, this definition of uncertainty implicitly requires the measurement scientist to
consider all the effects that might influence the measurement result. These effects obviously
include the causes of random variation from one measurement to the next over the
measurement timescale. But it is also essential to consider sources of bias during the
experiment, and very often, these generate larger effects than can be observed by repeated
measurement alone. That is, measurement uncertainty automatically asks for a range that
includes an allowance for both random and systematic effects.
5.1.6 To consider a simple analytical example, a simple measurement of concentration in a
solid will typically involve extraction of material, weighings, volumetric operations and
perhaps spectrometry or chromatography. Repeated measurement will show a spread of
values due to random variations in these operations. But all analysts know that extraction is
rarely complete, and, for a given material, that failure to extract material will lead to a
consistently low result. While good analytical practice always attempts to reduce such effects
to insignificance, some bias will remain. In expressing the uncertainty about the value of the
measurand, then, the analyst must take into account the reasonable possibility of bias from

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 Uncertainty of measurement

such causes. (Usually, this is done by considering such information as the range of analyte
recoveries observed on reference materials or from spiking experiments.)
5.1.7 The same considerations apply in the case of sampling. It is well known that different
samples taken from a bulk material will often show real variation in value, which is clear
from repeated measurement. It is also well known that sampling may be biased, for example
by differential removal of materials, inappropriate timing of sampling where temporal
fluctuations occur, or by access restrictions. These effects will influence the relationship
between the value of the measurand and the result that is observed. While good practice in
sampling is intended to reduce these effects to insignificance, a careful assessment of
uncertainty always considers the possibility of residual systematic effects.
5.1.8 Current guidance on measurement uncertainty [2] makes it clear that uncertainty of
measurement (Section 2.2 of reference [2]) is not intended to allow for gross error. This
would preclude, for example, mistakes caused by transcription errors or gross misuses of the
measurement protocol. Sampling can, however, produce high levels of uncertainty (e.g. 80%
of the concentration value), simply through the routine application of an accepted
measurement protocol to a highly heterogeneous material. Even when procedures are
nominally correct, there will also be slight variations in the actual procedures due to the
ambiguity in the measurement protocols, and the minor adaptations that are made to protocols
in real-world sampling situations. Whether these high levels of uncertainty lead to
unacceptable levels of reliability in the decisions that are based upon them, depends upon a
rigorous evaluation of fitness for purpose (see Section 16).

5.2 Specification of measurand

5.2.1 When an end-user is presented with a concentration result quoted for a bulk sample
in the form xU, they will very naturally interpret that interval as including the range of
values attributable to the concentration in the sampling target (e.g. a batch of material).
Implicit in this view is the idea that the measurand is the (true) concentration (of the analyte)
in the batch of material, and that the uncertainty includes any necessary allowance for
heterogeneity in the bulk. The analyst, by contrast, might refer to the concentration in the
laboratory sample analysed, implicitly ruling out the variation between laboratory samples.
Clearly, one viewpoint includes the effects of sampling, while the other does not. The effect
on the uncertainty can, of course, be very considerable. In metrological terms, this distinction
arises because the two views are considering different measurands. One is considering
concentration in the sampling target, the other concentration in the laboratory sample.
Another example might be contaminant concentration at a factory outlet at the time of
sampling, compared to the average contaminant concentration over a year.
5.2.2 These ambiguities in interpretation can be avoided only by careful specification of
the measurand. It is clearly necessary to state the quantity (mass, length, concentration etc.). It
is equally important to be clear on the scope of the measurement, by including information on
factors such as the time, location, or population to which the measurement result will be
assumed to apply. Some particular instances of measurand specification and their implications
for uncertainty estimation are discussed below.
It is never possible to avoid all ambiguity in implementing the wording of the sampling
protocol.
5.2.3 When a composite sample is taken by the combination of several increments from
across a sampling target, and analysed as a single primary sample, that single determination of
analyte concentration provides an estimate of the value of the measurand (i.e. the average
composition of the target), as discussed briefly in Section 4. The uncertainty on this single

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 Uncertainty of measurement

value reflects the uncertainty in the estimate of the measurand value. In contrast, if several
independent primary samples are taken from the target, each analysed once, and the mean
value calculated, this mean value will also be an estimate of the value of the measurand.
However, the uncertainty will not be that of the measurement (expressed as standard
deviation, s), but the standard error of the mean value (expressed as s/n). This later
uncertainty on the mean can be reduced by taking more primary samples,c whereas the
uncertainty on the measurement cannot.

5.3 Error, precision and uncertainty

5.3.1 Uncertainty is related to other concepts, such as accuracy, error, trueness, bias and
precision. Other guidance discusses the relationships in some detail [1, 2]. However, it is
worth repeating some of the important differences:
Uncertainty is a range of values attributable on the basis of the measurement result and
other known effects, whereas error is a single difference between a result and a true (or
reference) value
Uncertainty includes allowances for all effects that may influence a result (i.e. both
random and systematic errors); precision only includes the effects that vary during the
observations (i.e. only some random errors).
Uncertainty is valid for correct application of measurement and sampling procedures, but,
as noted in Section 5.1.8, it is not intended to make allowance for gross operator error

5.4 Sampling and physical preparation as sources of uncertainty of measurement

5.4.1 The act of taking a sample introduces uncertainty into the reported measurement
result wherever the objective of the measurement is defined in terms of the analyte
concentration in the sampling target and not simply in the laboratory sample.
5.4.2 Sampling protocols are never perfect in that they can never describe the action
required by the sampler for every possible eventuality that may arise in the real world in
which sampling occurs. The location in space (or time) for the taking of a sample is rarely
specified exactly (e.g. to the nearest millimetre or second). The sampler has to make such
decisions (ideally on objective criteria), but as heterogeneity is inevitable (in space or time)
such decisions will affect the estimated concentration. An appreciation of these sources of
uncertainty is important in the design and implementation of methods for the estimation of
uncertainty. When duplicate samples are taken, for example, taking them at exactly the same
place and time may not reflect the uncertainty of the measurement that really exists. This will
be discussed further in the description of methods of estimation (Sections 6 to 10), and in the
various worked examples (Appendix A).
5.4.3 Heterogeneity always gives rise to uncertainty. If the sampling target were perfectly
homogeneous then this contribution would be zero, but nearly all materials are heterogeneous
to some extent at some scale. If the test portion is a few micrograms, then nearly all material
will be heterogeneous and the sampling step will contribute to the uncertainty in the
measurement of an analyte concentration. Heterogeneity can be quantified in a separate
experiment, but if the aim is to estimate the analyte concentration in the larger sampling
target, then this heterogeneity is just one cause of measurement uncertainty (as discussed in
Section 4.2).

c
Assuming that the samples are random and independent, and assuming zero bias.

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 Uncertainty of measurement

5.4.4 Similar arguments can be made for the uncertainty that arises in the processes of
physical preparation (e.g. transportation, preservation, comminution, splitting, drying, sieving,
homogenisation) that happen after the act of sampling and before the chemical preparation of
the test sample (Figure 1). Each step can introduce errors from a range of mechanisms, such
as loss of analyte, loss of fine particles, or contamination from equipment or previous
samples. The methods employed, and training given, should aim to reduce these errors to a
minimum. In addition, however, procedures are required to estimate the uncertainty that all of
these steps, when applied in practice, generate in the final measurement value.

5.5 Sources of uncertainty

5.5.1 Uncertainty arises from a variety of sources, and these have been categorised in
different ways. For example, the Eurachem Uncertainty Guide identifies eight major
categories of effects that are important in estimating uncertainty have been identified [3], of
which the first two are sampling and sample preparation. Specific effects identifiable in these
two categories are shown in Table 1. A modelling approach might use these effects as the
basis for a mathematical model. Alternatively, sampling theory identifies eight distinct
sources of error in sampling (Table 2); each of these can also be reduced to a variety of causal
factors, which in turn can be used in various modelling approaches. A further alternative
approach is to consider all of the steps in the measurement process (Figure 1) as sources of
uncertainty that make some contribution to the uncertainty of the final measurement. In this
Guide, the simplest study designs treat uncertainty as arising from four classes of effect
(Table 3), and the classes are treated as sources of uncertainty in a simple statistical model;
this is consistent with the grouping of uncertainty sources explicitly suggested in reference
[3]. In its simplest form, this categorisation can be reduced to two categories: sampling
uncertainty and analytical uncertainty.
5.5.2 The important feature of each of these different classifications is that each is intended
to ensure that, however they are grouped and evaluated, all practically important effects are
taken into account in estimating the uncertainty. As long as this requirement is met, any
categorisation scheme may be applied to the estimation of uncertainty. The categorisation
schemes listed in Table 2 and Table 3 cover all practically important effects.
5.5.3 Each different categorisation of sources will generally lead to a different study
design, and very often to fundamentally different methods of evaluation of uncertainty
contributions. This results in substantially independent estimates of uncertainty via different
approaches. As noted elsewhere [5], grossly different estimates of uncertainty for the same
system suggest that at least one study methodology is in error. This forms the basis of a check
on the validity of an approach. Where practicable, therefore, comparison of uncertainty
estimates arising from independent evaluation approaches is recommended as a means of
validating particular estimates and of assessing the validity of different approaches.

5.6 Heterogeneity as a source of uncertainty

5.6.1 IUPAC currently define both homogeneity and heterogeneity as The degree to
which a property or constituent is uniformly distributed throughout a quantity of material.
(see Appendix B for definitions). So defined, heterogeneity is among the most important
factors contributing to uncertainty associated with sampling. Increments from different
locations in the sampling target will have different concentrations of analyte in a
heterogeneous material and there will be a sample-to-sample variation in analyte
concentration usually visible as a contribution to the observed variation of results. In
general, the exact dependence of concentration on location is unknown, so no correction can

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be made. This results in uncertainty in any given result or, in general, any average of such
results.
5.6.2 IUPAC note, as an addendum to the above definition, that The degree of
heterogeneity (the opposite of homogeneity) is the determining factor of sampling error. The
note is a good indication of the importance of heterogeneity in sampling. There are other
sources of error and uncertainty in the general operation of sampling; for example, cross-
contamination and imperfect stabilisation of samples, either of which can result in (unknown)
bias or additional variability. Yet heterogeneity and its effects such as random variability
and selection bias remain the largest problem in properly managed sampling and will
generally be the most significant source of uncertainty.
5.6.3 An alternative definition of homogeneity is sometimes used for particulate material,
which, if it consists of particles of different materials, cannot ever be homogeneous in the
sense defined by IUPAC. In this context, a mixture in which the probability of selection of
different types of particle is constant throughout the sampling target may be termed
homogeneous to denote that the expected concentration in would be the same in a sample
taken at any point in the material. Even here, however, it must be recognised that the
particulate nature of the material leads to sample-to-sample variation due to slightly different
composition of the increments actually taken; heterogeneity, as defined by IUPAC, still has
an effect under these circumstances, and consequently still contributes to the uncertainty.

Table 1: Some sources of uncertainty in sampling and sample preparation, adapted

from reference [3]
Sampling Sample preparation
- Heterogeneity (or inhomogeneity) - Homogenisation and/or sub-sampling
- Effects of specific sampling strategy (e.g. effects
random, stratified random, proportional - Drying
etc.) - Milling
- Effects of movement of bulk medium
- Dissolution
(particularly density selection)
- Extraction
- Physical state of bulk (solid, liquid, gas)
- Contamination
- Temperature and pressure effects
- Derivatisation (chemical effects)
- Effects of sampling process on
composition (e.g. differential adsorption - Dilution errors
in sampling system) - (Pre-)Concentration
- Transportation and preservation of - Control of speciation effects
sample

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Table 2: Sources of sampling uncertainty in sampling theory*

Source Description
Fundamental sampling error (FSE) A result of the constitutional heterogeneity
(the particles being chemically or
physically different)
Grouping and segregation error (GSE) A result of the distributional heterogeneity
Long-range point selection error (PSE1) Trends across space or over time
Periodic point selection error (PSE2) Periodic levels across space or over time
Increment delimitation error (IDE) Identifying the correct sample to take.
Considers the volume boundaries of a
correct sampling device
Increment extraction error (IXE) Removing the intended sample. Considers
the shape of the sampling device cutting
edges
Increment and sample preparation error Contamination (extraneous material in
(IPE) sample):
Losses (adsorption, condensation,
precipitation etc.):
Alteration of chemical composition
(preservation):
Alteration of physical composition
(agglomeration, breaking of particles,
moisture etc.):
**Involuntary mistakes (mixed sample
numbers, lack of knowledge, negligence):
**Deliberate faults (salting of gold ores,
deliberate errors in increment delimitation,
forgery etc.)
Weighting error (SWE) The result of errors in assigning weights to
different parts of an unequal composite
sample
*This classification follows that of Gy [17] and others (discussed further in Section10).
** Excluded from uncertainty estimates as gross errors [2].
Table 3: Uncertainty contributions in the empirical approach
Process Effect class*
Random (precision) Systematic (bias)
Analysis Analytical variability Analytical bias (combined effect
(combined contribution of of bias sources)
random effects)
Sampling Sampling variability Sampling bias
(dominated by heterogeneity (combined effect of selection bias,
and operator variations) operator bias etc.)
*The differentiation of random from systematic effects can depend on the context. A systematic effect in
measurements by one organisation (e.g. analytical bias) can also be considered a random effect when viewed in
the context of the consensus value from an inter-organisational proficiency test.

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 Approaches to uncertainty estimation

PART 3 Estimation of measurement uncertainty including

sampling

6 Approaches to uncertainty estimation

6.1 There are two broad approaches to the estimation of uncertainty. One of them,
described as empirical, experimental, retrospective, or top-down, uses some level of
replication of the whole measurement procedure to give a direct estimate of the uncertainty
for the final result of the measurement. This approach is called the empirical approach in
this Guide. The second, variously described as modelling, theoretical, predictive or
bottom-up, aims to quantify all of the sources of uncertainty individually, and then uses a
model to combine them. It will accordingly be referred to as the modelling approach. These
approaches are not mutually exclusive. The empirical method can be adapted to estimate
contributions to uncertainty from one or more effects or classes of effect. Both approaches can
usefully be used together to study the same measurement system, if required. The
applicability of the two approaches varies between the different materials to be sampled.
6.2 The approach taken in this Guide is to describe in detail the empirical approach,
which has the widest applicability to the broadest range of measurement systems and
applications (e.g. gaseous, liquid and solid). Modelling approaches are described for
particular situations to which they apply (e.g. particulate solids). Advice will also be given on
how a combination of these different approaches can be used to give more reliable and cost-
effective estimates of uncertainty in a range of measurement systems. This dual approach is
intended to enable a user of the Guide to select the most appropriate method of uncertainty
estimation for their particular situation. (Section 1 provides guidance on selection of the
approach.)
6.3 Reference [5] notes that the modelling approaches and the type of empirical study
used in collaborative trial are extremes of a continuum:
Note, however, that observed repeatability or some other precision estimate is very
often taken as a separate contribution to uncertainty even in the [modelling] approach.
Similarly, individual effects are usually at least checked for significance or quantified
prior to assessing reproducibility. Practical uncertainty estimates accordingly often
use some elements of both extremes.
In referring to either extreme, therefore, it is important to be aware that these are extremes and
that many practical estimates involve elements of both approaches.
6.4 The overall objective of any approach is to obtain a sufficiently reliable estimate of
the overall uncertainty of measurement. This need not necessarily require all of the individual
sources of uncertainty to be quantified, only that the combined effect be assessed. If, however,
the overall level of uncertainty is found to be unacceptable (i.e. the measurements are not fit
for purpose) then action must be taken to reduce the uncertainty. Alternatively, the
uncertainty may be unnecessarily small, in which case there may be justification for
increasing the analytical uncertainty, and thereby decreasing the cost of analysis. Methods for
modifying uncertainty are discussed in Appendix E. At this stage, however, it is essential to
have information on which general part of the measurement procedure is causing the
dominant contribution to the overall uncertainty, and it may then be necessary to evaluate
individual effects. The advantage of detailed early study is that this information is already
available; the disadvantage is that it is costly to obtain and may prove unnecessary if
uncertainty is acceptable. Planners should accordingly consider the level of detail required in

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 The measurand and general conduct of studies of sampling uncertainty

an uncertainty estimate, taking account of the probability of requiring detailed information for
further development.

7 The measurand
7.1 In the following discussion, it is assumed that the measurand is an average value
representing the composition of the whole sampling target, and that the measurand is to be
estimated through a process of sampling and analysis. This relates to the specification of the
measurand (Section 5.2) and the definition of the sampling target (Section 4.1).

8 General conduct of studies of sampling uncertainty

8.1 Analytical work should be undertaken under an appropriate quality system, including
validated analytical methods, proficiency testing, internal quality control and external
assessment where appropriate. Validation procedures should include all the steps normally
undertaken within the laboratory (including any sub-sampling of test samples), and should
include checks on bias using certified reference materials, or other methods, for the estimation
of analytical bias. Note that the uncertainty estimation methods described in this Guide can
also be applied to the estimation of uncertainties associated with sub-sampling.
8.2 Laboratories undertaking the chemical analysis should report the concentration
estimates exactly as found; in particular, values must not be censored, truncated or reported as
less than a reporting limit, whether below the limit of detection (LOD) or below zero.
Failing to report negative or sub-LOD observations will result in an underestimate of the
uncertainty.

9 Empirical approach
9.1 Overview
9.1.1 The empirical (top-down) approach is intended to obtain a reliable estimate of the
uncertainty, without necessarily knowing any of the sources individually. It relies on overall
reproducibility estimates from either in-house or inter-organisational measurement trials. It is
possible to describe the general type of source, such as random or systematic effects, and to
subdivide these as those arising from the sampling process or the analytical process. Estimates
of the magnitude of each of these effects can be made separately from the properties of the
measurement methods, such as sampling precision (for random effects arising from
sampling) or analytical bias (for systematic effects arising from chemical analysis). These
estimates can be combined to produce an estimate of the uncertainty in the measurement
result. This approach is illustrated in detail in Examples A1, A2, A3 and A4.

9.2 Uncertainty sources

9.2.1 It is possible to consider uncertainty of measurements to arise from four broad
sources of error. These four sources are the random errors arising from the methods of both
the sampling and analysis, and also the systematic errors arising from these methods. These
errors have traditionally been quantified as the sampling precision, analytical precision,
sampling bias and the analytical bias respectively (Table 4). If errors from these four sources
are quantified, separately or in combinations, it is possible to estimate the uncertainty of the
measurements that these methods produce. Methods for the estimation of three of the four
errors are well established. Sampling and analytical precision can be estimated by duplication

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 Empirical approach

of a proportion (e.g. 10%) of the samples and analyses respectively. Analytical bias can be
estimated by measuring the bias on well-matched certified reference materials, and assuming
that this bias represents that present for the test material, or by taking it directly from the
validation of the analytical method.
9.2.2 Procedures for estimating sampling bias include the use of a reference sampling
target [10, 24] (the sampling equivalent of a reference material). Alternatively they utilise
measurements from inter-organisational sampling trials, in which the sampling bias
potentially introduced by each participant is included in the estimate of uncertainty based on
the overall variability (Section 9.5). Although some of the components of uncertainty
associated with systematic effects may be difficult to estimate, it may be unnecessary to do so
if there is good evidence that systematic effects are small and under good control. Such
evidence may be qualitative, as in prior knowledge of the chemical or physical nature of the
sampling target, or quantitative, such as information, for example from prior measurements
on complete batches. (See Examples A3 and A4, Appendix A.)

Table 4: Estimation of uncertainty contributions in the empirical approach

Process Effect class
Random (precision) Systematic (bias)
Analysis e.g. duplicate analyses e.g. certified reference materials
Sampling Duplicate samples Reference sampling target,
inter-organisational sampling trial
Four classes of effects that contribute to the uncertainty of measurements, and methods for their estimation.

9.3 Statistical model for the empirical estimation of uncertainty

In order to design experimental methods to estimate uncertainty using the empirical approach
it is necessary to have a statistical model describing the relationship between the measured
and true values of analyte concentration. This random effects model considers a single
measurement of analyte concentration (x), on one sample (composite or single), from one
particular sampling target:
x = X true + sampling + analysis

where X true is the true value of the analyte concentration in the sampling target (i.e. equivalent
to the value of the measurand). For example, this could be the total mass of the analyte in the
target divided by the total mass of the target. The total error due to sampling is sampling and the
total analytical error is analysis

In an investigation of a single sampling target, if the sources of variation are independent, the
measurement variance 2meas is given by,
2 meas = 2 sampling + 2 analytical
where 2 sampling is the between-sample variance on one target (largely due to analyte
heterogeneity), and 2 analysis is the between-analysis variance on one sample.
If statistical estimates of variance (s2) are used to approximate these parameters, we get

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 Empirical approach

s 2 meas = s 2 sampling + s 2 analytical

The standard uncertainty (u) can be estimated using smeas , which is therefore given by

u = smeas = s 2 sampling + s 2 analytical Equation 1

Variance caused by physical sample preparation can be included into the sampling variance,
or expressed as a separate term if required.
In a survey across several sampling targets, which is recommended for the estimation of
sampling uncertainty (Section 9.4.2), the model needs to be extended to
x = X true + target + sampling + analysis

where the additional term target represents the variation of concentration between the targets
and has variance 2 betweent arg et . Appropriate ANOVA generates estimates of the variances
2 betweentarget , 2 sampling and 2 analysis , and the uncertainty is estimated exactly as before, using
Equation 1.
The total variance 2 total , given by
2total = 2betweentarget + 2 sampling + 2 analytical
is also a useful parameter in assessing fitness for purpose; this is discussed further in section
16.2. For practical purposes the population variances are replaced by their estimates s2 to
give
s 2 total = s 2 betweentarget + s 2 sampling + s 2 analytical Equation 2

9.4 Empirical estimation of uncertainty

9.4.1 Four types of method are applicable to the estimation of uncertainty using the
empirical approach (Table 5). A fifth variographic method is described briefly in Section 9.6.
The main method described further in this Guide is the duplicate method (#1). If one
sampler uses several sampling protocols in Method #2, any bias between the protocols can be
detected. If multiple samplers all apply one protocol (Method #3, which is equivalent to a
collaborative trial in sampling CTS, or method performance test), then bias between
different samplers can be detected and included in the uncertainty estimate. If multiple
samplers apply different protocols that are selected to be the most appropriate for the stated
objective, in their professional opinion (Method #4, which is equivalent to a sampling
proficiency test SPT), then any sampling bias introduced by either the sampling protocol, or
the sampler, can be detected and included in the estimate of uncertainty.

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 Empirical approach

Table 5: Four empirical methods for estimating combined uncertainty including

sampling
Method Method Samplers Protocols Component estimated
# description (persons) Sampling Analytical
Precision Bias Precision Bias
1 Duplicates Single Single Yes No Yes No1
2 Protocols Single Multiple Between protocols Yes No 1
3 CTS Multiple Single Between samplers Yes Yes 2
4 SPT Multiple Multiple Between protocols Yes Yes 2
+between samplers

1
Analytical bias information may be obtained by including certified reference materials in the analytical run (see
Example A2, Appendix A).
2
Analytical bias is partially or completely included in collaborative exercises where multiple laboratories are
involved.

9.4.2 The duplicate method is the simplest and probably most cost-effective of the four
methods described in Table 5. It is based upon a single sampler duplicating a small proportion
(i.e. 10%, but no less than eight targets) of the primary samplesd [11, 12]. Ideally the
duplicates are taken from at least eight sampling targets, selected at random to represent the
typical composition of such targets. If only one target exists, then all eight duplicates can be
taken from it, but the uncertainty estimate will only be applicable to that one target. The
duplicated samples are taken by repeating the same nominal sampling protocol, with
permitted variations that reflect the ambiguity in the sampling protocol and the effect of
small-scale heterogeneity of the analyte of interest on the implementation of that protocol. For
example, in a W design for collecting a composite sample over a bay of lettuces, the initial
starting point and orientation of the W is altered for the duplicate sample; for a grid design,
again, starting point and orientation are altered (ExampleA1, Appendix A). The duplicate
samples are obtained using a single sampling protocol and by a single person (sampler). Both
of the duplicated samples are subject to physical preparation resulting in two separate test
samples. Duplicate test portions are drawn from both of the test samples and analysed in
duplicate (i.e. duplicate chemical analysis). This system of duplicated sampling and chemical
analysis is known as a balanced design (Figure 2). Note that the duplicate method does not
include any contribution from sampling bias, which must be either assumed to be negligible,
or estimated separately using, for example, multiple samplers, multiple protocols and/or inter-
organisational sampling trials as in the other three methods.
Note: Although the duplicate method is generally described in terms of a single sampler and
protocol, the same design can be used with different samplers to incorporate the between-
operator contribution to uncertainty (equivalent to Method #3).

d
A higher level of replication can be used, but duplication is usually the most effective form of replication in
sampling studies. It is better to take duplicates from 12 sample targets, than take triplicates from eight targets, for
example, as although each estimate of the uncertainty of sampling (ssampling) has a lower standard error, the
estimate is based on a smaller proportion of the entire population of sample targets, and is therefore less
representative. The minimum number of eight duplicates is required to provide sufficiently reliable estimates of
the uncertainty [12].

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 Empirical approach

Figure 2: A balanced design

10% of targets in whole
Sampling survey n 8
target between-target variance

Sample 1 Sample 2 between-sample variance

sampling precision s samp

-between-analysis variance
Analysis 1 Analysis 2 Analysis 1 Analysis 2 analytical precision s anal

Balanced experimental design for empirical estimation of uncertainty (i.e. two-stage nested design), using the
duplicate method.

9.4.3 The test portions are then chemically analysed anonymously by an appropriate
analytical method under repeatability conditions (e.g. distributed randomly within an
analytical batch). If estimates of the analytical portion of the measurement uncertainty have
been made independently by the laboratory, this will be useful for comparison with estimates
made by this method. Variance caused by physical sample preparation can be included into
the sampling variance by having independent preparation on each of the sample duplicates.
Alternatively this variance can be estimated separately by inserting an extra level of
replication in the experimental design (Appendix D).
9.4.4 The balanced design proposed here will only give the repeatability standard deviation
of the analytical measurements. In order to estimate the other part of the analytical
uncertainty, an allowance has to be made for potential analytical bias. The limitations of this
approach, and a worked example, are given in Section 6 of Example A2. One alternative is to
ask the measuring laboratory for the repeatability and measurement uncertainty, and then to
check that the repeatability obtained in this study is similar to that claimed by the laboratory.
If this is the case, we can use the measurement uncertainty given by the lab as u(analytical)
(normally U/2). A second alternative is to use the estimation of analytical bias made from the
well-matched certified reference materials contained in the analytical batch. This bias
estimate can then be combined with the repeatability to obtain the measurement uncertainty
[1, 30].

9.5 Calculation of uncertainty and its components

9.5.1 The random component of the uncertainty can be estimated by applying analysis of
variance (ANOVA)e or range calculationf to the measurements of concentration on the
duplicated samples. The estimation is based upon the model described in Section 9.3, applied
to whatever measurement protocol that is being employed (with its specified number of
sample increments and analytical replicates).

e
There is often a small proportion (i.e. <10%) of outlying values in the frequency distributions of the analytical,
within-sample and between-sample variability. This requires the use of some method of down-weighting the
effect of the outlying values on classical ANOVA, such as the use of robust statistical methods. This gives a
more reliable estimate of the variances of the underlying populations. A fuller explanation of these methods is
given in the worked example in Appendix A1 (p40), A2 (p45).
f
See example in Appendix A2.

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 Empirical approach

9.5.2 The values of ssamp and sanal from the ANOVA are estimates of sampling precision
and analytical precision respectively. The random component of the measurement uncertainty
is calculated by the combination of these two estimates (Equation 1). The expanded
uncertainty, for approximately 95% confidence for example, requires this value to be
multiplied by a coverage factor of 2. The expanded uncertainty (U) is then calculated using
U = 2 smeas Equation 3
U can also be expressed relative to the reported value x and expressed in terms of a
percentage, as a relative expanded uncertainty U':
2 s meas
U ' = 100 % Equation 4
x
The relative uncertainty is more widely applicable than the standard uncertainty, as it does not
change appreciably as a function of concentration at values well above the analytical detection
limit (>10 times). Other coverage factors can be selected as appropriate. The improvement of
this estimate of uncertainty to include the systematic error from the chemical analysis is
discussed in Example A2, Appendix A.
The relative expanded uncertainty for the sampling or analysis alone can similarly be
expressed as
2 ssamp
U samp '= 100 %
x
and
2sanal
U anal ' = 100 %
x
9.5.3 Because the uncertainty of many measurement systems is dominated by
heterogeneity within the sampling target, the use of the simplest duplicate method often
gives a reasonably reliable estimate of uncertainty. Studies of environmental systems have
shown that between-operator and between-protocol effects are often much smaller than those
caused by heterogeneity [43]. Further information on selection of the most effective method
for uncertainty estimation is provided in Section 1. Examples of applications of the duplicate
method are given in Examples A1 and A2, Appendix A.
9.5.4 In addition to an initial single estimate of uncertainty for a particular sampling
protocol applied to a particular sampling target, routine application of the duplicate method
is also useful as a way of monitoring the ongoing sampling quality (Section 13). This can
allow for the effect on uncertainty of changes in the heterogeneity of the sampling target
between different applications of the same sampling protocol. Quantitative evidence of the
quality of sampling can then be gained, rather than relying solely on the assumption that
samples are representative, if taken by a correct protocol.

9.6 Alternative empirical methods of uncertainty estimation

9.6.1 Variography has also been suggested as a further empirical means of estimating the
uncertainty of measurement from the combined sources of sampling and analysis [13]. It is
particularly useful in situations where there is large-scale spatial and/or temporal variation in
contaminant concentration that can be quantified and modelled. This is the case for some
instances of rock and soil geochemistry, and in emission control of (e.g. waste water), when

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 The modelling approach

large numbers (n>100) of evenly distributed samples have been taken. Further guidance on
the principles and application of variography for this purpose, with a case study, is available
[8].

10 The modelling approach

10.1 Cause-and-effect modelling
10.1.1 The modelling approach, often colloquially known as bottom-up, has been
described for measurement methods in general [2], and applied to analytical measurements
[1]. It initially identifies all of the sources of uncertainty, quantifies the contributions from
each source, and then combines all of the contributions, as a budget, to give an estimate of the
combined standard uncertainty. In the process, the measurement method is separated into all
of its individual steps. This can usefully take the form of a cause-and-effect, or fish-bone,
diagram [3]. The uncertainty of measurement generated by each of these steps is estimated
independently, either empirically or by other methods. The combined uncertainty is then
calculated by combining the uncertainty from all of the steps by appropriate methods. This
approach is well established for analytical methods [1], but has only recently been applied to
the process of sampling [13, 14]. For particulate systems, sampling theory uses a similar
approach to identifying seven types a sampling error. One of these errors (fundamental) is
estimated using an equation based on a detailed knowledge of the individual particles being
sampled, as discussed in the next section (and Example A5, Appendix A).

10.2 Sampling theory for estimation of uncertainty

10.2.1 Sampling theory has been proposed as an appropriate method for estimating
uncertainty from sampling [15]. This approach relies on the use of a theoretical model, such
as that of Gy. Pierre Gy has developed a complete sampling theory described in many
publications [6, 16, 17, 18, 19, 20], including its latest developments [19]. Figure 3 shows
Gys classification of sampling errors. Most sampling errors, except the preparation errors,
are due to the material heterogeneity, which can be divided into two classes: 1) constitution
heterogeneity (CH), and 2) distribution heterogeneity (DH). Both heterogeneities can be
mathematically defined and experimentally estimated. Constitution heterogeneity refers to the
fact that all natural materials are heterogeneous, that is, they consist of different types of
particles (molecules, ions, grains). The distribution is heterogeneous if the particles are not
randomly distributed in the sampling target (or lot) to be investigated.

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 The modelling approach

Figure 3: Classification of sampling errors in Gys sampling theory

Global
GlobalEstimation
EstimationError
Error
GEE
GEE

Total Sampling Error Total Analytical Error

TSE TAE

Point
PointMaterialization
MaterializationError
Error Weighting Error
PME
PME SWE

Increment
Increment Delimi-
Delimi- Increment
IncrementExtraction
Extraction Increment
Incrementand
andSample
Sample
tation
tationError
Error Error
Error Preparation
PreparationError
Error
IDE
IDE IXE
IXE IPE
IPE

Point Selection Error

PSE

Long Range Periodic

Point Selection Error Point Selection Error
PSE1 PSE2

Fundamental Grouping and

Sampling Error Segregation Error
FSE GSE

GEE = TSE + TAE

TSE = ( PSE + FSE + GSE ) + ( IDE + IXE + IPE ) + SWE
* incorrect sampling errors are indicated by shaded boxes, and are excluded from the estimates of uncertainty
10.2.2 The classification of errors of sampling forms a logical and useful framework for
designing and auditing sampling procedures. Those that are central to the estimation of
uncertainty (e.g. FSE in Figure 3) are discussed below, and others (SWE, PSE and GSE) in
Appendix C.
10.2.3 The total determination error, which Gy calls the global estimation error (GEE), is
the sum of the total sampling error (TSE) and total analytical error (TAE). The components of
TSE can be divided into two major groups: 1) errors of incorrect sampling, 2) errors of correct
sampling. Some incorrect sampling errors arise from what the GUM [2] refers to as gross
errors, and as such would be excluded from estimates of uncertainty. The errors of correct
sampling occur within good practice and can be considered for inclusion within estimates of
uncertainty following the GUM approach [2].
10.2.4 Incorrect sampling errors arise from sampling equipment and procedures that do not
follow the rules of sampling correctness defined in the sampling theory. In Figure 3 these
errors are shown in shaded boxes. Increment delimitation error (IDE) is error that is generated
if the shape of sample is not correct. For example, from a process stream the correct sample is
a complete slice of equal thickness cut through the process stream. The sampling device
should be designed so that it can extract the intended sample profile (i.e. all constituents have
an equal chance to end up in the sample). Otherwise sample or increment extraction error

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 The modelling approach

(IXE) is created. Sample preparation errors (IPE) have several potential causes listed in Table
2, two of which are excluded as gross errors by the GUM definition.
10.2.5 Incorrect sampling errors have the following properties in common: 1) they create
sampling bias and increase the total variance in an unpredictable way, 2) they are
circumstantial and, therefore, any attempt to estimate them experimentally is normally not
useful, because it is expensive and the results cannot be generalised. The correct way is to
minimise or eliminate these errors by carefully auditing the equipment and procedures, by
replacing structurally incorrect devices and procedures with those that follow the rules of
sampling correctness, and by sufficient training of sampling personnel. Only if this technical
part is correctly executed does the theoretical part of uncertainty evaluation have predictive
value. However, sampling uncertainty estimation and quality control may alert users to
procedures that are not behaving correctly.
10.2.6 Correct sampling errors are shown in the lower part of Figure 3. When the
incorrect sampling errors are eliminated these errors can be modelled and used for estimating
the uncertainty of sampling. The fundamental sampling error is among the most important and
will be considered further here; others are discussed in Appendix C2.
10.2.7 Fundamental sampling error (FSE) is the minimum error of an ideal sampling
procedure. Ultimately it depends on the number of critical particles in the samples. For
homogeneous gases and liquids it is very small but for solids, powders and particulate
materials, especially at low concentrations of critical particles, fundamental error can be very
large. If the lot to be sampled can be treated as a one-dimensional object, fundamental
sampling error models can be used to estimate the uncertainty of the sampling. If the lot
cannot be treated as a one-dimensional object, at least the point selection error has to be taken
into account, when the variance of primary samples is estimated. If the sample preparation
and size reduction by splitting are carried out correctly, fundamental sampling error models
can be used for estimating the variance components generated by these steps. If the
expectance value for the number of critical particles in the sample can be estimated easily as a
function of sample size, Poisson distribution or binomial distribution can be used as sampling
models to estimate the uncertainty of the sample. In most cases the fundamental sampling
error model can be used.
10.2.8 If the material to be sampled consists of particles having different shapes and size
distributions it is difficult to estimate the number of critical particles in the sample. An
equation can be used to estimate the relative variance of the fundamental sampling error:
1 1
r2 = Cd 3 Equation 5
MS ML
where
a
r = = relative standard deviation of the fundamental sampling error
aL
a = absolute standard deviation (in concentration units)
aL = average concentration of the lot
d = characteristic particle size = 95% upper limit of the size distribution
MS = sample size
ML = lot size

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 The modelling approach

C is a sampling constant that depends on the properties of the material sampled; C is the
product of four parameters:
C = f gc Equation 6
f = shape factor (see Figure 4)
g = size distribution factor (g = 0.25 for wide size distribution and g = 1 for uniform particle
sizes)
= liberation factor (see Figure 4). For materials where the particles are completely
x
L
liberated, = 1. For unliberated material an empirical equation, = , is used, where
d
values of x ranging from 0.5 to 1.5 have been suggested.
c = constitution factor and can be estimated if the necessary material properties are available
by using:
2
aL
1
a
c= c + 1 L m Equation 7
aL

Here aL is the average concentration of the lot, the concentration of the analyte in the
critical particles, c the density of the critical particles and m the density of the matrix or
diluent particles.
10.2.9 If the material properties are not available and they are difficult to estimate, the
sampling constant C can always be estimated experimentally. Certified reference materials,
for example, are a special group of materials for which the sampling constant can be
estimated from existing data.
10.2.10 An example of how the fundamental sampling error model can be used in practice is
given in Example A5, Appendix A.

Figure 4 Estimation of factors for the estimation of fundamental sampling error.

d d d d

f= 1 f= 0,524 f= 0,5 f= 0,1

default in most cases

d

L=d
L
x
L =1
=
d

The particle shape factor f (upper part), and liberation factor for unliberated material (lower
left) and liberated material (lower right). L is the liberation size of the critical particles.

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 Responsibility for quality of sampling

PART 4 Management issues

11 Responsibility for quality of sampling

11.1 The implications of regarding sampling as an integral part of the measurement
process are far reaching, and include management issues. The rigour that is applied to
assessing and improving the quality of activities within the analytical laboratory should be
applied equally to the sampling procedures. The responsibility for the quality of the whole
measurement process should ultimately rest with one organisation, and responsibilities for
different parts of the process must additionally be defined. Similarly, one body should take
responsibility for estimating the measurement uncertainty, based on information from all
participants. This organisation can then inform all of the participants of the contributions
arising from the main steps in the measurement procedure.

12 Selection of uncertainty estimation approach

12.1 The empirical (top-down) and modelling (bottom-up) approaches each have their
advantages in certain circumstances. These should be considered in selecting the approach for
a particular sampling exercise.
The empirical approach includes all sources of uncertainty, without the scientist having to
know their identity in advance. For example, it is automatically applicable to the
particular contaminants and mineralogy present at a geological site. The calculations do
not require any prior knowledge of the nature of the material (e.g. grain size, analyte
speciation, degree of heterogeneity). It is relatively quick and easy to apply practically
(especially for the duplicate method). There are at least four options available to allow
progressively more accurate (and more expensive) estimates of uncertainty, as appropriate
(Table 5). Some of these methods can allow for systematic error (such as sampling bias)
within the estimate of uncertainty. Sampling proficiency tests and reference sampling
targets are still in the early stages of development, but already show considerable promise
for this application.
Among the disadvantages of the empirical approach is that it does not necessarily quantify
any of the individual components of uncertainty (although this knowledge can be added
with limited resolution). It is not based on a theoretical model of particulate sampling, but
this may be an advantage in applications to materials that are not particulate in form (e.g.
gaseous, liquids, biota). The empirical approach only gives an approximate value of
uncertainty, which is assumed to be constant over the target, but this is also true of the
modelling approach. Extreme values in the replicate measurements may lead to an
overestimate of the uncertainty value, which is not representative of most measurements.
This effect can be minimised, however, by the use of robust statistics.
The principal advantage of the modelling approach is that it allows the largest source of
uncertainty to be readily identified, if it was in the model. It gives a transparent method
showing which components of uncertainty have been considered in the summation of
uncertainty. Finally, where prior information is available, modelling approaches can be
less costly than extensive experimental studies.
The disadvantages of the modelling approach include that the theoretical predictions of
uncertainty may require detailed prior measurements of the mineralogy, grain size and
analyte speciation of the material to be sampled (e.g. soil), and how these vary across the

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 Selection of uncertainty estimation approach

target. Idealised assumptions have to be made therefore about the make up of the material
(e.g. mineralogy, grain size and analyte speciation). The modelling approach using
sampling theory requires estimates or assumptions about eight types of sampling error,
and also how these might vary across the target. Both theoretical and empirical
approaches can be relatively time consuming and therefore expensive to implement.
Generic estimates may be too general and not reflect the specific circumstances at any
particular sampling target. Further, not all of the sources of uncertainty might be
identified, leading to an underestimate of the total uncertainty.
On balance, therefore, the empirical methods tend to be more generally applicable across a
wide range of types of material, and do not depend as heavily on prior knowledge of the
system or all of the sources of uncertainty. This will make them less time consuming, and
therefore less costly to apply, which is particularly valuable in one-off testing of different
sampling targets. The modelling approaches, by contrast, lead to a more detailed assessment
of individual known sources of uncertainty and are more appropriate when developing a long-
term sampling scheme for a specific well-characterised application.

13 Quality control of sampling

13.1 Relationship between validation and quality control
13.1.1 Once an uncertainty that makes the measurements fit for purpose has been
established, evaluation of the sampling and analytical procedures proposed to meet those
purposes can be undertaken. Two evaluation tools are needed for this purpose: validation and
continual quality control.
13.1.2 Validation comprises a one-time estimation of the uncertainty components
determined under conditions expected to be encountered in the routine use of the procedures.
The validation may be done generically for the sampling method (initial validation) or site
specifically for the method used on site to the selected target (on-site validation). Initial
validation is used when sampling is done as a one-time campaign (spot sampling, e.g.
contaminated site investigation) and on-site validation is repeated at intervals (repeated
sampling, e.g. time or flow-proportional sampling of waste water). In short, validation
demonstrates what can be achieved and, if that conforms to the fitness-for-purpose
requirement, the procedures are deemed suitable for routine use. The methods for validation
are described in the previous chapters of this Guide.
13.1.3 Validation alone cannot ensure that routine results are indeed fit for purpose,
however. Routine or site specific conditions may differ from those prevailing during
validation, either systematically or occasionally. This is especially true for sampling, where
the larger part of the uncertainty component often stems from the heterogeneity of the target,
that is, where the degree of heterogeneity may vary markedly from one target to the next. This
is also true when a sampling method is applied at different sites. These circumstances
emphasise the need for an ongoing internal quality control that includes sampling, to ensure
that conditions prevailing at validation (and therefore the expected uncertainty attached to the
results) are still applicable every time that the sampling and analytical procedures are
executed. The combined use of validation and quality control is shown in Table 6.

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 Quality control of sampling

Table 6: Illustration of the combined use of validation and quality control of sampling
One method used at One method used
many sites repeatedly at one site
Validation Initial validation yielding On-site validation yielding
generic performance data the performance data for
the specific target
Quality control Extensive quality control Spot quality control
with site specific verifying the performance
verification of generic data consistency over time
performance data

13.1.4 The need for internal quality control of sampling is not widely recognised at present,
and methods for executing it are not well established, except in some specialised areas such as
geochemical prospecting [21]. Specific suggestions for sampling quality control are given for
some environmental sampling matrices in [22]. However, no new principles are involved;
with minor qualification, the principles of internal quality control of analysis are applicable to
sampling [23, 24, 25]. Moreover, the methods used in validation are, with some
simplification, applicable to internal quality control. The reason for the simplification is that
validation needs to provide a good estimate of uncertainty, while quality control merely needs
to demonstrate consistency, over space and time, with the uncertainty established at
validation.

13.2 Methods of internal quality control of sampling

13.2.1 The focus of interest is almost exclusively the precision aspect. Bias is difficult to
address in validation and almost impossible in internal quality control. The reference target,
the conceptual equivalent in sampling of a certified reference material [26], is rarely available.
Moreover, it is not fully useful: we need to see whether results for individual sampling targets
are fit for purpose, not whether unbiased and reproducible results can be obtained on a
possibly unrepresentative reference target.
13.2.2 The principal tool is replication. This is minimally executed by taking two samples
from each target by a complete (and suitably randomised) duplication of the sampling
protocol. Each sample is analysed once and the difference between the results
D = x1 x2 calculated. If the validated uncertainties of sampling and analysis are u s and u a

respectively, the combined standard uncertainty is u meas = (u 2

s )
+ u a2 . Consequently, a one-
sided range control chart can be constructed with a control limit (at the 95% confidence
interval) of 2.83u meas and an action limit (at the 99% confidence interval) of 3.69u meas [25]
(Figure 5). An out-of-control value of d shows that the result should be scrutinised as possibly
unfit for purpose. Such a result is not diagnostic and may stem from a disturbance in either
sampling or analysis; the latter should be detected by standard methods of analytical quality
control.

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 Reporting and interpreting uncertainty

Figure 5: Example of an R-chart for quality control of sampling

R-Chart: NNH4

2.2
2.0
1.8
1.6
1.4
1.2

g/l
1.0
0.8
0.6
0.4
0.2
0.0
14-Oct 20-Oct 26-Oct 29-Oct 5-Nov 17-Nov 24-Nov 30-Nov 10-Dec
Date of analysis

For the construction of this R-chart see the Nordtest Guide [25].

13.2.3 The data from quality control can also be used to update sampling method precision
as obtained in method validation using the same methods, ANOVA or relative difference
calculations.
13.2.4 In some instances, the extra cost of duplicate sampling can be eliminated by use of
the SAD (Split Absolute Difference) method in which the normal number of increments to be
combined as the sample is segregated at random into two equal sized sub-sets, each of which
is processed and analysed separately [27, 28]. The difference between such results has an
uncertainty of 4u s2 + 2u a2 if conditions applying to validation are maintained. This again
could be used to define an action limit in a one-sided control chart.

14 Reporting and interpreting uncertainty

14.1 Introduction
14.1.1 It is crucial to ensure that reports are clear as to the measurand being reported. In
particular, it is important to be clear whether the result and its uncertainty apply to a single
test portion, a laboratory sample, the whole of a sampling target (e.g. a bulk material), or a
series of targets. Using the principles of the GUM [2] and previous Eurachem/CITAC Guides
[1], uncertainty will initially be estimated in the form of a standard uncertainty, u, which
includes due allowance for all effects which may reasonably influence the result. Uncertainty
may be quoted in this form without change. However, it is often convenient to report in other
forms for increased confidence or for wider applicability. It is essential to note any limitations
in the estimate of uncertainty, such as the exclusion of sampling bias or other neglected
effects. The following paragraphs describe the most important issues, and give some guidance
on their interpretation.

14.2 Expanded uncertainty, U

14.2.1 The standard uncertainty u applied to a result in the form xu, and associated with a
normal distribution, describes an interval including only about 68% of the area of the
distribution. This is usually taken to indicate that there is a greater than 32% probability of the
measurand value being outside this interval. This is considered insufficient confidence for
most practical applications. It is therefore normal practice to apply a suitable multiplier to the
standard uncertainty so that the quoted interval includes a greater proportion of the dispersion.

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 Reporting and interpreting uncertainty

Conventionally, this multiplier, usually designated k, is referred to as the coverage factor, and
the product ku=U is referred to as the expanded uncertainty.
14.2.2 The choice of k is discussed in considerable detail in other publications [1, 2].
However, the key principles are:
k should be chosen to reflect an approximate confidence interval for the particular
distribution.
If a particular distribution is known to be applicable, it is used. Otherwise, a normal
distribution is considered a reasonable assumption where the dominant contributions to
uncertainty are all normally distributed or there are several approximately equal
contributions from arbitrary distributions. With this assumption, k is typically based on the
value of Students t for an appropriate (two-tailed) level of confidence and number of
degrees of freedom.
In the modelling approach, the number of degrees of freedom is formally derived from the
degrees of freedom for contributing uncertainties according to a published formula [1, 2],
or approximated from the number of degrees of freedom for the dominant contribution
[1]. More commonly, the number of degrees of freedom is assumed to be sufficiently
large to justify a choice of k=2 for approximately 95% confidence.
For most practical purposes, k=2 is considered acceptable, and is sometimes mandatory [29].
However, it is important to state the value of k used, and the approximate level of confidence
that k implies, when reporting expanded uncertainty.

14.3 Relative uncertainty statements

14.3.1 It is often found that the standard uncertainty from sampling increases approximately
proportionally with the value of the result. Under these circumstances, it is often most
practical to quote the uncertainty in a relative form, such as a relative standard deviation (u/x)
or percentage interval using Equation 4 (e.g. 10%). The relative value quoted is usually
based on an estimate of uncertainty for one or more representative results, but is applicable
over a greater range of concentration values.
14.3.2 It is important not to extrapolate a simple relative standard deviation to zero
concentration, as uncertainty does not normally disappear entirely at very low levels and the
proportionality assumption is no longer valid. More general approaches to these situations can
either specify a range of concentration over which the relative uncertainty value applies [25],
or else express the uncertainty as a function of concentration [1, 42].

14.4 Contributions to uncertainty

14.4.1 The exact steps that are included in each contribution to the measurement uncertainty
need to be stated. Depending on the method of estimation employed, the details of the
experimental design, and the person to whom the information is intended, it is possible to
quantify some specific components of the measurement uncertainty. For example, the
experimental design in Figure 2 will give separate estimates of two components called
sampling and analysis. When the details of this particular implementation of the design are
examined it becomes evident that uncertainty from physical sample preparation is included
within the general title of sampling, whereas that from chemical preparation is included
within analysis. If required, it is possible to insert a further level of duplication of physical
preparation within the experimental design to estimate the separate contribution which that
particular step introduces [30]. The exact steps that are included in each contribution to the
measurement uncertainty need to be documented. For less experienced users of analytical

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 Cost of estimating uncertainty from sampling

measurements, it may be better to report one value for the whole uncertainty of the
measurement, stating in a footnote which sources have been considered.

14.5 Applicability of estimates

14.5.1 Recalling the discussion of specification of the measurand (Section 5.2) it is crucial
to ensure that reports are clear as to the measurand being reported. As observed in Section
14.1.1, it is particularly important to state clearly whether the result and its uncertainty apply
to a single test portion, a laboratory sample, the whole sampling target, or to a series of
targets. Unlike estimates of uncertainty for analytical measurements, it is very probable that
the same sampling protocol will produce measurements with different levels of uncertainty
from sampling when it is applied to a new sampling target. New estimates will be required for
substantially different targets, particularly when there is reason to suppose that the degree of
heterogeneity has changed.

14.6 Interpretation of uncertainty statements against limits

14.6.1 Results are often compared with tolerances or regulatory limits in order to assess
compliance with a requirement. In making such comparisons, it is important to take
uncertainty into account. A full discussion is beyond the scope of the present Guide; more
detailed discussion will be found in references [1] and [31]. The basic principles are:
Decide whether the decision requires proof of compliance, proof of non-compliance, or a
shared risk approach, and set an appropriate level of confidence.
For proof of compliance, the result and its uncertainty interval must be entirely within the
permitted range.
For proof of non-compliance, the result and its uncertainty interval must be entirely
outside the permitted range.
For shared risk approaches, set a range for acceptable measurement results based on the
permitted interval, adjusted to provide a specified probability of false acceptance and false
rejection rates. Recent guidance gives useful details of the procedure [32].
For regulatory purposes, it is important to consult the specific regulations applicable, as no
general guidance can currently cover all cases. For example, it is generally considered unsafe
to pass material that is not proven compliant, dictating a proof of compliance approach.
Criminal prosecution in most countries, however, requires clear proof of non-compliance and
in these circumstances (e.g. blood alcohol prosecutions) it is normal practice to seek proof of
non-compliance at high levels of confidence.

15 Cost of estimating uncertainty from sampling

15.1 It would seem logical to consider the total budget for validation and quality control
of sampling to be judged together against the costs that will arise from erroneous decisions
based on inadequate estimates of uncertainty. It is recognised that implementing uncertainty
estimation will increase the overall costs of measurement. Applying the duplicate method, for
example, can increase the cost of sampling by up to 10%, and the analysis by 30% (i.e. three
additional analyses are required for applying the balanced design to 10% of the sampling
targets. An unbalanced experimental design may be performed, in which only one of the
duplicate samples is analysed twice, if suitable statistical treatment is performed. This
increased cost can be justified, however, by the additional information gained and the reduced

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 Judging fitness for purpose of measurements using uncertainty

potential losses from incorrect decisions that might have been made without knowledge of the
uncertainty (Section 16).
15.2 It is more difficult to evaluate general costs for the other methods of uncertainty
estimation. Inter-organisational sampling trials require the expenses of at least eight different
participants (to obtain an acceptable reliability [11]), and are therefore likely to be
significantly higher than those for the duplicate method. Modelling methods will require
detailed information about the material being sampled. For some materials that are relatively
consistent over many batches these values may be generally applicable, and therefore make
this approach more cost-effective than empirical methods that take larger numbers of extra
measurements on each batch. This discussion must therefore include the extent to which the
uncertainty value for a particular protocol/material combination is estimated at a preliminary
validation, and how much the value is continually monitored and/or updated by an ongoing
sampling quality control scheme (Section 1). It would seem logical to consider the total
budget for validation and quality control of sampling to be judged together against the costs
that will arise from erroneous decisions based on inadequate estimates of uncertainty.

16 Judging fitness for purpose of measurements using uncertainty

16.1 A proper understanding of uncertainty from sampling must be embedded in the
broader perspective of fitness for purpose. Three approaches have been suggested for setting
fitness-for-purpose criteria. The first approach is to set an arbitrary limit on the maximum
value of uncertainty that is considered acceptable. This approach has been widely applied in
the analytical sector, where a target relative uncertainty has been applied (e.g. 10%). The
problem with this approach is that it does not necessarily relate to intended purpose for which
the user requires the measurement.
16.2 The second approach is to compare the variance generated by the measurement
(sampling and analysis) to the variance of the measurements between the different sampling
targets. There are many situations where the objective of the measurements is to compare
concentrations between different targets, such as in mineral exploration where the objective is
to locate a target with significantly higher concentration of an element of interest (e.g. gold).
One application of this approach, for example, sets the fitness-for-purpose criterion so that the
measurement variance does not contribute more than 20% to the total variance (defined in
Equation 2) [33].
16.3 The third, and most generally applicable, approach to judging the fitness for purpose
of measurements, is consider the effect of the measurement on its ultimate purpose. All
analytical measurement is undertaken to support a decision. A decision can be either correct
or incorrect. An incorrect decision involves extra costs, and an incorrect decision is more
likely if the uncertainty is higher. Consider, for example, the manufacture of a material
against a specification of a maximum acceptable level of an impurity.g Each batch of material
is analysed to determine the level of the impurity. A false positive result has the outcome
that the batch of material is discarded or reworked unnecessarily to reduce the apparently
unacceptable level of impurity. A false negative result means that a defective batch is
released to the customer, a situation that may require financial compensation. Both of these
situations are more likely to occur if the uncertainty is higher. This seems to suggest that the
measurement should be undertaken so that the uncertainty is the smallest that can be

g
This concept is equally applicable to situations where materials have regulated minimum analyte
concentrations, in which case the terms false compliance and false non-compliance are applicable.

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 Judging fitness for purpose of measurements using uncertainty

achieved. However, reducing the uncertainty of a measurement result requires rapidly

escalating costs. A useful rule here is that, where random variation dominates the uncertainty,
the cost of a measurement is inversely proportional to the square of the uncertainty; a
reduction in uncertainty by a factor of 2 calls for an increase in expenditure by a factor of 4.
16.4 The true cost of a decision is the sum of the measurement costs and the excess costs
of incorrect decisions. From the above we can see that this sum has a minimum value at some
particular level of uncertainty (Figure 6), and this uncertainty is the definition of fitness for
purpose.

Figure 6: Schematic loss functions dependent on uncertainty of measurement

Line A shows the costs of measurement. Line B shows costs of incorrect decisions. The sum
of these two lines (the total cost shown by the highest line) shows a minimum cost at point C,
which is the uncertainty that can be regarded as fit for purpose.

16.5 The optimal apportionment of resources between sampling and analysis is also a
matter of costs. Even an elementary consideration (excluding costs) shows that the
uncertainties of sampling and analysis should be roughly balanced. For example, if the
uncertainties of sampling and analysis are 10 and 3 units respectively, the overall uncertainty
of measurement is 10 2 + 32 = 10.4 . The overall uncertainty is hardly affected by a reduction
of the uncertainty of analysis: if it is reduced to (say) 1 unit, the overall uncertainty is reduced
to 10 2 + 12 = 10.05 , an inconsequential change. A more sophisticated approach takes into
account the different costs of analysis and sampling. If the unit costs of sampling and analysis
are A and B for the same specific level of uncertainty, the optimum ratio of sampling
uncertainty u samp to analytical uncertainty u anal is given by
14
usamp A
= .
uanal B

This ratio provides the minimum expenditure for a given overall uncertainty of 2
usamp + uanal
2

or, alternatively, the minimum uncertainty for a given expenditure [34].

Methods for modifying uncertainty from sampling are discussed in Appendix E, although
operating at minimum total cost is not always achievable or necessary.

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 Implications for planning sampling and measurement strategies

17 Implications for planning sampling and measurement strategies

17.1 Expertise and consultation
As Section 4 shows, the sampling and analytical processes cover a range of activities.
Different parts of the process are frequently allocated to different staff, who may have very
different knowledge of the objectives and, more importantly, differing knowledge of the
effect of different parts of the process. In general, all of those involved will have good
knowledge of some part of the process, but few are able to advise on the complete process. It
is therefore important that sample planners involve analytical chemists and experienced
sampling technicians where possible in planning sampling. It is also prudent to include
statistical experts in most circumstances (see below). Decision makers (i.e. business managers
and those acting on the results of sampling activities) should be involved in planning for new
applications, and regulators should also be consulted where a protocol is intended to support
regulation.
Although the principles of this Guide are widely applicable, expert statistical guidance is
always valuable and should be considered essential in some circumstances. These include:
where the observed or expected frequency distributions are not normal, for example where
the results contain more than 10% outliers, or where the results show markedly
asymmetric distributions;
where large financial or social consequences depend on a reliable estimate of uncertainty;
where confidence intervals are needed on the estimates of uncertainty or, for more
complex sampling plans, on the measurement results;
where the sampling strategy is more complex than simple random sampling with
replicated measurements, for example in implementing stratified sampling.

17.2 Avoiding sampling bias

The methods described in this Guide are suitable for establishing the variability of sampling,
but only the more complex methods can begin to assess uncertainties associated with possible
bias in sampling. For this reason, close attention should be paid to minimising potential
sources of bias. These include possible bias associated with differential sampling due to
particle size, density or flow-rate; bias in selection of sampling points; the effect of different
sampling equipment etc. Specific expertise in sampling methodology should be sought unless
these factors can be demonstrated to be adequately controlled or are completely specified by
an established sampling protocol.

17.3 Planning for uncertainty estimation

Sampling exercises should always make provision for at least some replicated samples and
measurements in order to assess the uncertainty of the results.

17.4 Fitness-for-purpose criteria

Planning should ideally begin with the establishment of clear fitness-for-purpose criteria,
taking into account the relative costs and uncertainties of sampling and analysis where they
are known or can reasonably be determined in advance. Section 16 provides guidance on how
analytical and sampling effort can be optimised.

UfS:2007 Page 32
 Implications for planning sampling and measurement strategies

17.5 Use of prior validation data

The main uncertainties associated with analytical measurements are often estimated during, or
on the basis of, analytical method validation, a process which is carried out prior to bringing
the method into use. Consideration accordingly needs to be given as to whether the variability
found as part of the sampling experiment should replace, inform, or simply serve as a check
on, the analytical measurement uncertainty assessed using prior information. In considering
this issue, it should be noted that the variability observed during a relatively short series of
analyses is rarely sufficient as an estimate of uncertainty. Long-term studies are generally
more reliable. It is accordingly safer to rely on prior validation data unless the observed
variation is significantly higher.
Uncertainties associated with sampling variability can themselves be estimated in advance,
particularly where a long-term sampling programme is to be planned and implemented. Under
these circumstances, it is usually prudent to obtain an initial estimate of sampling uncertainty.
Ongoing studies can then serve as a check on continuing validity of the uncertainty estimate,
for example by applying internal quality control principles as discussed in Section 13.

17.6 Acceptability of sampling uncertainty

Before reporting measurements, it should be evaluated whether they are acceptable and in
accordance with the quality objectives set for the whole uncertainty and its sampling
component, probably based on some fitness-for-purpose criterion, prior to the measurements.

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 Appendix A: Examples

Appendix A: Examples

Introduction
The most effective way to explain the methodologies described in the main text of this Guide
is to show worked examples. These examples are not intended to cover all circumstances, but
to show how the general principles can be applied to a variety of situations across a range of
different sectors. These include food (production and retail), animal feed, and environment
(soil and water). The examples are all structured using the same basic format, so as to aid
comprehension and comparability.

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 Example A1

Example A1: Nitrate in glasshouse grown lettuce

Measurand Uncertainty estimation

Analyte/ Unit1 Sector/ Sampling Purpose Design Statistics
Technique Matrix target(s)
Nitrate/Hot mg kg-1 Food/ 1 bay of Uncertainty Empirical Robust
water as received Lettuce Iceberg total - duplicate ANOVA
extraction and lettuce measurement, method
determination grown under sampling and
by HPLC glass analytical

1 Scope
Estimate the measurement uncertainty, and contributions from sampling and analysis, for
routine monitoring of glasshouse grown lettuce, using a standard sampling protocol.

2 Scenario and sampling target

Nitrate is essential for plant health; however, there are concerns for human health associated
with eating elevated levels of nitrate. The concentrations of nitrate in lettuce are regularly
monitored in line with EC requirements. Concentration estimates are made for each bay of
up to 20,000 lettuce heads, and the result for each bay used individually in assessing
conformance with the relevant Regulation. Each bay is accordingly considered a sampling
target, rather than individual heads of lettuce. In order to make a reliable comparison of the
measured nitrate concentrations against the European regulatory threshold [35] (4500 mg kg-
1
), an estimate of the measurement uncertainty is desirable.

3 Sampling protocol
The accepted protocol for this purpose specifies that one composite sample is prepared from
10 heads of lettuce harvested from each bay of lettuce [36]. The lettuces are selected by
walking a W shape or five-point die shape through the bay under investigation. This protocol
is applied to all bays regardless of the size. Samples were taken in the morning and
transported to the contracted analytical laboratory in ice-packed cool boxes to arrive within 24
hours of sampling.

4 Study design duplicate method (Section 9.4.2)

The minimum of eight targets were selected for inclusion in the uncertainty estimation
protocol. For each of these bays a second 10-head sample was taken (S2) in addition to the
routine sample (S1). This duplicate sample was taken in a way that represented the variation
that could occur due to the ambiguities in sampling protocol, for example positioning of the
origin of the W design, and its orientation.

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 Example A1

Figure A1.1: Example of the 'duplicate method'

Example of how the duplicate method can be applied. Using the W design as an example, the protocol
stipulates the design but not the position or orientation. The W is equally likely to start on the left or
the right. Ten heads are taken along the line of the W to create a composite sample for one target.

5 Sampling and analysis in the laboratory

Primary samples were frozen on receipt at the laboratory. A lettuce (increment) from each 10-
head sample was cut into four equal quarters and two quarters retained. This was repeated for
each of the 10 increments in the sample. The resultant 20 quarters were place in a Hobart
processor and macerated to process a composite sample. Two analytical test portions (10 g)
were taken. Each test portion was extracted using hot water and the nitrate concentration was
determined by HPLC (ultra-violet detector). Quality control samples (spike recovery) were
analysed concurrently with the real samples. No significant analytical bias could be detected
and so bias correction was considered unnecessary for the resultant data. The raw
measurement values use for the estimation of uncertainty had appropriate rounding, and no
suppression of values less than either zero, or the detection limit.

6 Results
The best estimates of the nitrate concentration at each of the eight target locations are shown
in Table A1.1.
Table A1.1: Measurements of the concentration (mg kg-1) of nitrate in eight duplicated samples. The duplicate
samples are labelled S1 and S2. Likewise, duplicate analyses are labelled A1 and A2. Hence, DS1A2 (value
4754 mg kg-1) is analysis 2, from sample 1 from sampling target D

Sample
S1A1 S1A2 S2A1 S2A2
target
A 3898 4139 4466 4693
B 3910 3993 4201 4126
C 5708 5903 4061 3782
D 5028 4754 5450 5416
E 4640 4401 4248 4191
F 5182 5023 4662 4839
G 3028 3224 3023 2901
H 3966 4283 4131 3788

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 Example A1

Before applying statistical methods it is useful to inspect the data to ascertain the general
levels of variability. The analytical duplicates (e.g. BS1A1 and BS1A2) are generally within
300 mg kg-1 of each other, suggesting an analytical precision of less than 10%. The sample
duplicates (e.g. DS1 and DS2) agree less well, but generally differ by less than 20%.
However, one target (C) displays a greater difference, suggesting an outlying value.
Quantification of the random component of the measurement uncertainty and two of its main
components (sampling and analysis) was made using robust analysis of variance (RANOVA,
Appendix C3, with output in Figure.A1.2. Robust ANOVA was used here as outlying targets
are, in this relatively well-controlled environment, considered likely to be anomalies, rather
than reflecting the underlying population statistics, and as a precaution against analytical
outliers.
Note: Robust methods should not be used where apparent outliers arise as part of the typical
population of sampled increments or targets, unless the specific implementation allows for
non-normal distributions for part of the assumed error structure.

Figure A1.2
CLASSICAL ANOVA RESULTS
Mean = 4345.5625
Standard Deviation (Total) = 774.5296
Sums of Squares = 12577113 4471511 351320
Between-target Sampling Analysis
Standard Deviation 556.2804 518.16089 148.18063
Percentage Variance 51.583582 44.756204 3.6602174

ROBUST ANOVA RESULTS:

Mean = 4408.3237
Standard Deviation (Total) = 670.57617
Between-target Sampling Analysis Measurement
Standard Deviation 565.39868 319.04834 167.94308 360.5506
Percentage Variance 71.090791 22.636889 6.2723172 28.909209
Relative Uncertainty 14.474814 7.6193626 16.357719
(% at 95% confidence)
The output of ANOVA for data produced form a balanced experimental design (n = 8, Table A1.1). Both robust
and classical estimates are given for comparison. Standard deviation estimates are computed for between-
target (sbetween-target), within-target (ssamp) and within-chemical analysis (sanal). Results are in the same units of
concentration as the input data (i.e. mg kg-1 in this case).

Extracting the robust estimates from this output gives:

ssamp = 319.05 mg kg-1
sanal= 167.94 mg kg-1
Equation 1 can be used to calculate:
smeas = (ssamp2 + sanal2 ) = 360.55 mg kg-1

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 Example A1

This can be used as an estimate of the random component of the standard uncertainty (u).
The expanded relative uncertainty is given by Equation 3 as:
Umeas' = 200 * 360.55 / 4408 = 16.4% (of concentration value)
For the sampling alone, the expanded relative uncertainty (random component) is similarly
given by:
Usamp' = 200 * 319.05 / 4408 = 14.5%
For comparison the expanded uncertainty for the analytical contribution (random component)
is given by:
Uanal' = 200 * 167.94 / 4408 = 7.6%
This value is less than the normal limits set within internal analytical quality control (e.g.
10%).
The analytical recovery estimates were not statistically different from 100% recovery (i.e. no
analytical bias was detected). For this example, therefore, no additional allowance was made
for uncertainty associated with analytical bias.

7 Comments
This uncertainty estimate does not include any estimate of the possible sampling bias.

8 Assessment of the fitness for purpose of these measurements

The fitness-for-purpose criterion used initially is that based on the percentage of total variance
(Section 16.2). When using RANOVA the program computes how much the between-target,
within-target (or sampling) and analytical variance contributes (as a percentage) to the total
variance (Figure A1.2). For this study of nitrate in lettuce the maximum contribution to the
total variance is from between-target variability (71.1%). By combining the sampling (22.6%)
and analytical contributions (6.3%) it is clear that the combined measurement process
contributes 28.9% of the total variance. This is marginally greater than the ideal of 20%. Of
this measurement variance, sampling is the dominant factor, responsible for 78.2% of the
measurement variance.
Fitness for purpose may also be assessed using the optimised uncertainty (OU) methodology.
This method addresses fitness-for-purpose assessment with financial considerations (Section
16.3)[37]. In this case it can be shown that an increase from a 10-head to a 40-head composite
sample is required to achieve fitness for purpose (Appendix E, and [38]).

9 Reporting and interpretation

For each bay of lettuce (sampling target), the nitrate concentration of the 10-head composite
sample is compared to the threshold value (4500 mg kg-1). Each nitrate concentration should
be reported with the measurement uncertainty (16.4% of the measured value) Table A1.2. The
interpretation of whether each batch exceeds a threshold value, based upon its measurement
and associated uncertainty, depends on the wording of the appropriate regulation [32]

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 Example A1

10 Summary

Measurement uncertainty
Sampling Analytical Total
14.5% 7.6% 16.4%

Table A1.2
Sample Expanded
S1A1
target Uncertainty

A 3898 639.3
B 3910 641.2
C 5708 936.1
D 5028 824.6
E 4640 761.0
F 5182 849.8
G 3028 496.6
H 3966 650.4
The nitrate concentrations associated with S1A1 (routine sample) are shown with the associated measurement
uncertainty (calculated from U = 16.4%). As an example, Target F has a value of the measurand (or true value)
between 4332 mg kg-1 and 6032 mg kg-1.

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 Example A2

Example A2: Lead in contaminated top soil

Measurand Uncertainty estimation

Analyte/ Unit Sector/Matrix Sampling Purpose Design Statistics
Technique target(s)
Total lead mg kg-1 Environmental/ 100 targets Uncertainty Empirical Robust
/ICP-AES dry Top soil each of area total - duplicate ANOVA
basis 30 m x 30 m, measurement, method
with depth of sampling and
0150 mm analytical

1 Scope
Estimate the measurement uncertainty, and contributions from sampling and analysis, at each
of 100 different sampling targets within one site, using a common sampling protocol.

2 Scenario and sampling target

An investigation was made of a 9-hectare site, as part of the assessment of the land for
potential housing development. The most important analyte element for human health risk
assessment was found to be lead. In order to compare the concentration of lead in the soil with
the national regulatory threshold limit (450 mg kg-1), an estimate of the lead concentration
and the measurement uncertainty was required for each of 100 sampling targets.

3 Sampling protocol
One hundred samples of top soil (nominal depth 0150 mm) were taken with a hand auger
(diameter 25 mm) at 100 locations. These locations were distributed on a regular grid with
sample spacing of 30 m (Table A2.1), and therefore each is intended to represent an area 30 m
by 30 m. The surveying was conducted with a measuring tape and compass.

4 Study design duplicate method (Section 9.4.2)

Ten of the samples (i.e. 10% of the total number), at randomly selected locations, were
sampled in duplicate using the balanced design (Figure 2). The duplicate samples were taken
at a distance of 3 m from the original sample, in a random direction. This aims to reflect the
ambiguity in the sampling protocol, the uncertainty in locating the sampling target (e.g. the
surveying error) and also the effect of small-scale heterogeneity on the measured
concentration within the specified target. Six soil certified reference materials (CRMs) were
selected for analysis to estimate analytical bias over a range of concentration.

5 Sampling and analysis in the laboratory

Primary samples were oven dried overnight at 60 C, disaggregated, sieved to remove particles
with a natural grain size greater than 2 mm (based upon the definition of soil). The sieved
samples (<2 mm) were all ground (95% < 100m) and mixed. Test portions of 0.25 g were

UfS:2007 Page 40
 Example A2

taken for dissolution with nitric and perchloric acids, prior to determination of lead by ICP-
AES [39]. The measurements were subject to full analytical quality control (AQC), and
corrected for reagent blank concentrations where these values were statistically different from
zero. The raw measurement values use for the estimation of uncertainty had no rounding or
suppression of values less than either zero, or the detection limit.

6 Results
The best estimates of the lead concentration at each of the 100 target locations are shown in
the format of a map (Table A2.1).

Table A2.1: Measured lead concentrations at each target on the sampling grid (mg kg-1), shown by the actual
coordinates used in the regular sampling grid (spacing 30 m). They show a high degree of variability between-
locations of roughly a factor of 10. The variability within 10 of these locations selected at random (i.e. A4, B7,
C1, D9, E8, F7, G7, H5, I9 and J5) was used for the estimation of uncertainty from sampling (Table A2.2). This
within-target variation is substantial (e.g. a factor of 2) but substantially less than the between-target variability.
Row A B C D E F G H I J
1 474 287 250 338 212 458 713 125 77 168
2 378 3590 260 152 197 711 165 69 206 126
3 327 197 240 159 327 264 105 137 131 102
4 787 207 197 87 254 1840 78 102 71 107
5 395 165 188 344 314 302 284 89 87 83
6 453 371 155 462 258 245 237 173 152 83
7 72 470 194 82.5 162 441 199 326 290 164
8 71 101 108 521 218 327 540 132 258 246
9 72 188 104 463 482 228 135 285 181 146
10 89 366 495 779 60 206 56 135 137 149

Four measurements from the balanced design for each of the 10 sample targets selected for
duplication were used for the estimation of uncertainty (Table A2.2). Visual inspection of the
data allows an initial qualitative assessment of the relative importance of the two sources of
measurement uncertainty. The low level of agreement between the concentration values from
some of the sample duplicates is indicative of a high level of sampling uncertainty (e.g. S1
compared to S2 for target D9). The agreement between the analytical duplicates (A1 and
A2) is however generally much better for most samples (< 10% difference) than that between
the sample duplicates.

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 Example A2

Table A2.2: Measurements of the concentration (mg kg-1) of a lead on 10 duplicated samples from the total of
100 targets in a survey of contaminated land (Table A2.1). The duplicate samples are labelled S1 and S2.
Likewise, duplicate analyses are labelled A1 and A2. Hence, D9S1A2 (value 702 mg kg-1) is analysis 2, from
sample 1 from sampling target D9. Values shown are rounded for clarity, and used for subsequent calculations,
but generally un-rounded values are preferable for these calculations.
Sample S1A1 S1A2 S2A1 S2A2
target
A4 787 769 811 780
B7 338 327 651 563
C1 289 297 211 204
D9 662 702 238 246
E8 229 215 208 218
F7 346 374 525 520
G7 324 321 77 73
H5 56 61 116 120
I9 189 189 176 168
J5 61 61 91 119

Quantification of the random component of the measurement uncertainty and two of its main
components (sampling and analysis) was made using robust analysis of variance (RANOVA),
with typical output shown in Figure A2.1. Robust statistics were selected to allow for the
outlying values that are evident in this data (e.g. target A4, sample duplicate D9S1/S2,
analytical duplicate B7S2A1/A2), and in most similar data sets (but see the Note in Example
A1, section 6). The estimates of uncertainty are averaged over the 10 targets, assuming that
the uncertainty is not varying significantly over this range of concentration. The uncertainty is
expressed in relative terms so that it is applicable over this range of concentration (Section
14.3).
Extracting the robust estimates from this output gives:
ssamp = 123.8 mg kg-1
sanal = 11.1 mg kg-1
Equation 1 can be used to calculate:
smeas = (ssamp2 + sanal2 ) = 124.3 mg kg-1
This can be used as an estimate of the random component of the standard uncertainty (u).
The expanded relative uncertainty is given by Equation 3, with a coverage factor of 2 as:
Umeas' = 200 * 124.3 / 297.3 = 83.63% (of concentration value)
For the sampling alone, the expanded relative uncertainty (random component) is similarly
given by:
Usamp' = 200 * 123.8 / 297.3 = 83.29%

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 Example A2

Figure A2.1: The output of ANOVA for data produced from a balanced experimental
design (n = 10, Table A2.2)
CLASSICAL ANOVA RESULTS
Mean = 317.79999
Standard Deviation (Total) = 240.19238
Sums of Squares = 1738031.9 370075.5 6473
Between-target Sampling Analysis
Standard Deviation 197.55196 135.43246 17.990274
Percentage Variance 67.646327 31.792678 0.5609926

ROBUST ANOVA RESULTS:

Mean = 297.30884
Standard Deviation (Total) = 218.48763
Between-target Sampling Analysis Measurement
Standard Deviation 179.67409 123.81386 11.144044 124.31436
Percentage Variance 67.62655 32.113293 0.26015487 32.373447
Relative Uncertainty 83.289726 7.4966113 83.626415
(% at 95% confidence)

Both robust and classical estimates are given for comparison. Standard deviation estimates are computed for
between-target (sbetween-target), within-target (ssamp) and within-chemical analysis (sanal). Results are in the same
units of concentration as the input data (i.e. mg kg-1 in this case).

For comparison the expanded uncertainty for the analytical contribution (random component)
is given by
Uanal' = 200 * 11.1 / 297.3 = 7.5%
This value is less than the typical limits set within internal analytical quality control (e.g.
10%).

Inclusion of analytical bias

The analytical bias was estimated as 3.41% (1.34%) using a linear functional relationship
[40] established between the measured values on the certified values of the six CRMs (Table
A2.3).
There is currently no consensus on the best way to combine random and systematic effects
into an estimate of uncertainty, although four options have been identified [30]. One option
[25] is to consider the estimated analytical bias (e.g. 3.41%) to be a typical value for
participants in an inter-organisational trial. If this bias, and its own uncertainty (1.34%) is
then added to the random component of the uncertainty (using the sum of squares) it will
increase the variance to that which would be found in such a trial. The logic of this approach
is that the extra uncertainty that is usually detected in inter-organisational trials is due to the
unsuspected bias within each organisation. Where an estimate can be made of the extra
variance caused by these biases between different laboratories, this can be added to the

UfS:2007 Page 43
 Example A2

random component within one organisation. In this case, the standard relative analytical
uncertainty is increased to 5.24% [ = (3.752 + 3.412 + 1.342)0.5]. The expanded analytical
uncertainty (10.48%) is then greater than the analytical target value of 10%, but it can also
usefully be compared with an independent estimate of the analytical measurement uncertainty
made within the laboratory. The expanded uncertainty for the whole measurement is thereby
increased to 83.95% [ = (83.292 + 10.482)0.5], which is practically identical to the purely
random component of 83.63%.

Table A2.3: Measured and certified lead concentration values for CRMs for the
estimation of the bias of the analytical method
CRM name Mean Standard Certified U on
(n=4) (mg kg-1) Deviation value certified
(mg kg-1) (mg kg-1) value
(95% conf.)

NIST2709 19.7 3.2 18.9 0.5

NIST2710 5352.0 138.0 5532.0 80.0
NIST2711 1121.4 14.7 1162.0 31.0
BCR141 34.4 3.9 29.4 2.6
BCR142 36.2 4.6 37.8 1.9
BCR143 1297.5 33.0 1333.0 39.0

7 Comments
This estimate of uncertainty does not make allowance for any undetected sampling bias
(Section 9.4.2). However, because the uncertainty is often dominated by the heterogeneity of
the sampling target, the extra uncertainty introduced by bias in sampling can often be
assumed to be insignificant by comparison (as shown for the analytical bias). Where the
highest quality of uncertainty estimate is required, due perhaps to potentially large financial
consequences from underestimating the uncertainty, it may be preferable to use one of the
more elaborate methods using multiple samplers and/or protocols (Table 5).
If the measurand (or true value) had been defined as the mean concentration of lead across the
whole site, the uncertainty would have had to include the contribution from the standard error
on the calculated mean value, expressed as stotal / n . For this example stotal is 403 mg kg-1, n
= 100 and the uncertainty on the mean (291.9 mg kg-1) is therefore 27.6% of the value, at 95%
confidence. This value can be calculated without knowing the individual contribution of the
uncertainty from either the sampling or the analysis, and is often dominated by sbetween-sample.

8 Assessment of the fitness for purpose of these measurements

Using the percentage of total variance method (Section 16.2), the output in Figure A2.1
attributes the percentage of the total variance ([standard deviation (total)]2 that is contributed
by between-target, sampling (within-target) and analysis (within-sample). In this particular

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 Example A2

example there is clearly a dominance of the between-target variance (67.6% of total

variance), although this is less than the ideal threshold of 80% (Section 16.2). Furthermore,
sampling dominates (32.11% of total variance) over chemical analysis (0.26% of total
variance) as a contributor to the measurement variance. Sampling variance (i.e. within-target)
is identified as the principal contributor (99.2%) of uncertainty in the measurement process in
this case (i.e. 100 * 32.11 / [32.11 + 0.26]).
The assessment of fitness for purpose of measurements in contaminated land investigation
using the optimised uncertainty method (Section 16.3), is described elsewhere [41].

9 Reporting and interpretation

Individual measurements of lead concentration reported for these targets should have attached
uncertainty values equal to 83.9% of the concentration value. This applies to all of these
measured values (Table A2.1), which are at least 10 times higher than the analytical detection
limit (estimated as 2 mg kg-1 in this case). In applications where this is not the case, it will be
necessary to express the uncertainty as a function of the concentration [42]. Furthermore, the
uncertainty on the mean measurements taken at the 10 targets where duplicate samples were
taken (e.g. those listed in Table A2.2) will have reduced uncertainty estimates of 59.3% (=
83.9/2).
Knowing the value of the uncertainty, it is also possible to make a probabilistic interpretation
of the level of lead contamination on the site [43].

10 Summary
Measurement uncertainty*
Sampling Analytical Total
83.3% 10.5% 83.9%
* with coverage factor of 2 (i.e. 95% confidence)

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 Example A3

Example A3: Dissolved iron in groundwater

Measurand Uncertainty estimation

Analyte/ Unit Sector/Matrix Sampling target Purpose Design Statistics
Technique
Dissolved mg l-1 Environment/ The groundwater Total Empirical Range
iron/ groundwater near one selected uncer- duplicates
ICP-AES monitoring well in tainty used in
a groundwater validation
body and quality
control

1 Scope
The scope is determination of the total uncertainty of the measurement of dissolved iron in a
sampling validation study and subsequent control of sampling uncertainty during monitoring.

2 Scenario and sampling target

A groundwater body that is an important drinking water resource for the city of rhus, the
second largest city of Denmark, has through surveillance monitoring been identified as at risk
for deterioration of the quality due to intensive drinking water abstraction. An operational
monitoring programme has been established in order to control the trend in water quality
development.
The groundwater body is in glacial outwash sand with Miocene sands and clays below and
glacial till above. The geology at the site is complicated with several local aquifers
(underground layer of water-bearing permeable rock, or permeable mixtures of
unconsolidated materials) and aquitards (geological formation of layers comprised either of
clay or on non-porous rock that restrict water flow from one aquifer to another). The
groundwater body as identified is 2 km x 2 km x 10 m, starting 2030 m below the surface.
The natural quality of the groundwater is anaerobic without nitrate, with sulphate and reduced
iron, but without hydrogen sulphide and methane. One of the threats to the groundwater body
is oxygen intrusion into the aquifer as the result of the water abstraction and concomitant
groundwater table drawdown.
In the groundwater body, nine wells had been sampled for chemical analysis during
surveillance monitoring, and six wells are now available for sampling. In the operational
monitoring plan, it was decided to aim at monitoring one well twice per year. The objective of
the operational monitoring was set to having a 95% probability of recognising a 20% quality
deterioration. It was decided to use dissolved iron as a target parameter that would be a
sensitive indicator of aquifer oxidation (decreasing iron concentration with increasing
oxidation) and with redox potential as supporting evidence. Oxygen, pH, electrical
conductivity and redox potential were used as on-line indicators of sampling stability and
sodium, calcium and chloride as general groundwater quality parameters. Only the two key
parameters, dissolved iron and redox potential, are discussed here.

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 Example A3

Meeting the monitoring objective requires a measurement uncertainty including both

sampling and analysis of not more than 10% (comparison of two means each for two samples,
95% confidence interval, two-sided test) corresponding to an expanded measurement
uncertainty of 20%. To ensure the compliance of the monitoring programme with the stated
objective, a sampling validation study was initially conducted including all wells available
and, based upon the results from this, a routine sampling quality control programme was set
up for implementation with the monitoring programme for the selected monitoring well.
The properties of the groundwater body were summarised based upon previous monitoring
activities (surveillance monitoring). Table A3.1 shows a summary for the two key parameters
including variability in time and space as well as measurement (sampling and analytical)
uncertainty.

Table A3.1: Key chemical parameters for nine wells to the groundwater body,
surveillance monitoring
Redox potential Dissolved iron
mV mg l-1
Mean -123 1.11
Relative standard deviation 27% 56%
Oxygen impact during sampling
Main cause of uncertainty Filtering
and on-line measurement

The chemical data suggest that the groundwater composition is quite uniform over time and
space with respect to the main components (data not shown, relative standard deviation 1.9
16%), whereas the variability is high for the redox parameters (oxygen, redox potential and
dissolved iron). The expected main causes of uncertainty are indicated in the table for the two
key parameters and the causes were controlled during sampling.

3 Sampling protocol
Sampling was done according to the rhus County groundwater monitoring protocol, with
permanent, dedicated pumps (Grundfos MP1) set in the middle of the screened interval of
each well. Pump rates were 12 m3 h-1 (well purging) with a 10% reduction just before
sampling. Two of the six wells were large-diameter abstraction wells equipped with high
yield pumps. These were pumped with 4060 m3 h-1 for well purging followed by pump rate
reduction just before sampling. During well purging, the development in water quality was
followed with on-line measurements of oxygen, pH, electrical conductivity and redox
potential until stable readings and then samples were taken. A field report was filled in during
the sampling, including pump yields and pumping times as well as water table measurements.

4 Study design empirical

The empirical approach was selected for study design in order to provide estimates of
heterogeneity in the groundwater body (between-target variation well-to-well and over time)
and measurement uncertainty, split to show sampling uncertainty and analytical uncertainty.

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 Example A3

4.1 Validation
The objective of the validation programme was to ensure that a measurement uncertainty
meeting the set quality objective could be obtained and to describe the components of
uncertainty in order to identify points of improvement, if required. The validation programme
was set up with sampling of six wells, two independent samplings per well and two sub-
samples per sample analysed, see Figure A3.1.

Figure A3.1 Design outline for validation

Ground-
water body

Well 1 Well 2 Well 3 Well 4 Well 5 Well 6

Sample 1 Sample 2

Analysis 1 Analysis 2

A total of 12 samples were taken and 24 sub-samples were sent for analysis in one sampling
round as a validation study.

4.2 Quality control

The objective of the quality control programme for the operational monitoring was to ensure
that measurement uncertainty did not increase over time during the monitoring. The quality
control programme was set up after careful evaluation of the results from the validation study.
Quality control was designed to include duplicate sampling, each with duplicate analysis, on
one of the two annual sampling occasions of the monitoring programme, see Figure A3.2. In
total, six sampling occasions with 12 samples and 24 sub-samples analysed were included in
the first phase of the quality control programme.

UfS:2007 Page 48
 Example A3

Figure A3.2 Design outline for quality control, shown for one sampling occasion
Groundwater body

Monitoring well

Sample 1 Sample 2

Analysis 1 Analysis 2

5 Sub-sampling and analysis

The sample pre-treatment and analytical set up for the two key parameters (redox potential
and dissolved iron) are shown in Table A3.2.

Table A3.2: Pre-treatment and analytical programme

Redox potential Dissolved iron
Pre-treatment On-line analysed On-line filtered, preserved with
nitric acid, laboratory analysed

5.1 Sub-sampling and sample pre-treatment

Duplicate on-line measurements/sub-samplings for laboratory analysis were done by taking
out split sample streams and treating each stream independently. This means that the
analytical uncertainty obtained with the duplicate design also included sub-sampling, pre-
treatment, such as filtering, and transportation. An estimate of the analytical uncertainty alone
could be obtained from the laboratory quality control data, see Section 5.3.
Samples were on-line filtered excluding oxygen through 0.45 m cellulose acetate membrane
filters and sub-samples were preserved in the field for metal analysis by acidification with
nitric acid. Sub-samples were stored in polyethylene containers in the dark at less than 10C
during transport to the analytical laboratory.

5.2 Field analysis

The sample stream was pumped through an on-line measuring array of a flow-through cell
with sensors set up in series. The sensor used for redox potential is described in Table A3.3.

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 Example A3

Table A3.3: On-line sensor used for redox potential measurements

Parameter Instrument Cell Instrument Calibration and
accuracy control
Redox potential pH 340 Sensolyt Pt 2 mV Daily service

No quality control was performed of on-line measurements in the field.

5.3 Laboratory analysis

Analyses were performed at an independent, accredited (ISO 17025) laboratory using
accredited methods subject to the required quality assurance and analytical quality control.
Methods and performance data from quality control are shown in Table A3.4.

Table A3.4: Methods and performance data from quality control for laboratory analyses
Method Within-series Between- Total Total Detection
repeatability series reproducibi expanded limit
reproduci- -lity uncertainty
bility
Iron ICP-AES 0.95% 4.2% 4.3% 8.6% 0.01 mg l-1

The certified reference material (CRM) VKI Metal LL2, nominal 0.200 mg Fe l-1 was used
for quality control with 101.9% recovery (mean for 92 control results).

5.4 Calculation methods

The replicate data were treated using the range method (ISO 3085). For comparison,
uncertainty estimates were calculated by analysis of variances (ANOVA) and robust ANOVA
(RANOVA) using ROBAN version 1.0.1 (Appendix C3).
The applied calculation methods are demonstrated in Section 7 (below). The range
calculations are easily done using standard spreadsheets, and an example can be downloaded
from http://team.sp.se/analyskvalitet/sampling/default.aspx.
The occurrence of systematic sampling errors was not assessed quantitatively, but the
consistency of the obtained results was used as a qualitative control of systematic errors. As
an example, if dissolved iron was found above 0.1 mg l-1 in the same sample as oxygen was
determined to be above 0.1 mg l-1, this would indicate a systematic sampling and/or pre-
treatment error. Similarly, redox potential and oxygen contents were checked to correspond in
order to control systematic errors.

6 Results
The data set from the validation study is shown in Table A3.8 for dissolved iron with the
range calculations. The calculations for redox potential in the validation study and for both
dissolved iron and redox potential during quality control were done similarly.
The data from the validation study (six different wells) using range calculations are shown in
Table A3.5.

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 Example A3

Table A3.5: Relative expanded uncertainty (%, coverage factor 2) for analysis, sampling
and between-target (between wells) as obtained during validation using range
calculations
Range calculations Analyses Sampling Between-target
Redox potential 5.2% 15% 14%
Dissolved iron 2.1% 10% 70%

For comparison, the statistical estimates are shown in Table A3.6 as obtained using ANOVA
and RANOVA.

Table A3.6: Relative expanded uncertainty (%, coverage factor 2) for analysis, sampling
and between-target (between wells) as obtained for dissolved iron during validation
using ANOVA and RANOVA calculations
Dissolved iron Analyses Sampling Between-target
ANOVA 1.6% 9.6% 70%
RANOVA 1.8% 9.9% 72%

The statistical estimates obtained with the range statistics during quality control (six sampling
occasions) are shown in Table A3.7.

Table A3.7: Relative expanded uncertainty (%, coverage factor 2) for analysis, sampling
and between-target (between occasions) as obtained during quality control using range
calculations
Analyses Sampling Between-target
Redox potential 18% 3.8% 23%
Dissolved iron 2.5% 3.6% 9.9%

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 Example A3

Table A3.8: Results and range calculations for the validation study, dissolved iron, basic data in bold, symbols used to describe
calculations only (T: target, S: sample, A: analysis, R: absolute differences, r: relative differences, n: numbers)
Well S1A1 S1A2 S2A1 S2A2 R1 = S1A1 S1A2 S1A1 + S1A2 r = R1 *100 R2 = S 2 A1 S 2 A2 S 2 A1 + S 2 A2 R2
h S1 = 1 S2 = r2 = *100 S = S1 + S 2 S1 S 2
number 2 S1 2 S2 2 r= *100
S
mg l-1 mg l-1 mg l-1 mg l-1 mg l-1 mg l-1 % mg l-1 mg l-1 % mg l-1 %
99.474 0.815 0.834 0.912 0.893 0.019 0.825 2.30 0.019 0.903 2.11 0.864 9.03
99.468 1.80 1.83 1.94 1.93 0.030 1.82 1.65 0.010 1.94 0.517 1.88 6.40
99.469 1.69 1.68 1.79 1.77 0.010 1.69 0.593 0.020 1.78 1.12 1.73 5.48
99.916 2.62 2.61 2.83 2.84 0.010 2.62 0.382 0.010 2.84 0.353 2.73 8.07
99.327 1.66 1.63 1.58 1.59 0.030 1.65 1.82 0.010 1.59 0.631 1.62 3.72
99.371 1.52 1.53 1.47 1.50 0.010 1.53 0.656 0.030 1.49 2.02 1.51 2.66
r = 7.413
1
r 2 = 6.750 S = 10.32 r = 35.36
nr1 = 6 nr 2 = 6 nr = 6 nr = 6
Analysis
rA =
r + r1 2 rA =
7.413 + 6.750
6+6
= 1.18 CV A =
rA i
CV A =
1.18
= 1.05
n r1 + n r 2 1.128 1.128
Sampling
rS + A =
r rS + A =
35.36
= 5.89 CVS + A =
rS + A
CVS + A =
5.89
= 5.22
CV S = CV S + A 2

CV A 2 j CVS = 5.22 2
1.05 2
2
= 5.17
nr 6 1.128 1.128 2
Between-target
ST + S + A =
S ST +S + A =
10.32
= 1.72
sT + S + A = s S k sT + S + A = 0.604
CVT + S + A =
sT + S + A
*100 CVT + S + A =
0.604
1.72
*100 = 35.1
nr 6 ST +S + A

CV S + A 2 5.17 2
2 CVS = 35.12 = 34.9
CVT = CVT + S + A 2
2

h
S1A1: sample 1 analysis 1.
i
The standard deviation can be obtained from the mean of relative differences between duplicate measurements by division with the statistical factor 1.128.
CV A 2
j
The sum of relative variances is CVS + A 2 = CVS 2 + with the factor on CV A 2 due to the mean of duplicate analyses being used.
2
k
s: standard deviation with n-1 degrees of freedom as obtained from most standard calculators and spreadsheets.
UfS:2007 Page 52
 Example A3
No groundwater samples had measurements of dissolved oxygen and dissolved iron
above 0.1 mg l-1, and the low redox potential measured (-110 to -200 mV) is
consistent with the absence of oxygen (<0.1 mg l-1) and the high dissolved iron
concentrations (0.92 to 2.8 mg l-1).

7 Comments
Overall, the validation data show that the variability in the aquifer (between-target)
was dominating the total uncertainty for dissolved iron, whereas sampling and
between-target uncertainties were of the same size for dissolved iron. Analytical
uncertainties were small (25%), and for dissolved iron were comparable to the
repeatability obtained in laboratory quality control (expanded uncertainty 2.1% as
compared to 1.9% respectively). If different wells were sampled, the sampling
uncertainty was 1015%.
For dissolved iron measured during validation, the use of ANOVA and RANOVA
calculations did not provide statistical estimates more than slightly different from
those obtained with the simple range calculations.
In the quality control scheme of monitoring, the variability between sampling
occasions (between-target, 9.9%) was dominating the total uncertainty for parameters
analysed as laboratory analysis (dissolved iron, 2.5% uncertainty), whereas the
analytical uncertainty (18%) was almost as important as the between-target
uncertainty (23%) for on-line measurements (redox potential). The reason for the
large contribution from on-line measurements is that during quality control, duplicate
on-line measurements were done with two different instruments in contrast to the
validation study done with one single instrument for both duplicate measurements.
Accordingly, the analytical uncertainty (instrument to instrument variation) for redox
potential was considerably larger in the quality control (18%) than in the validation
study (5.2%). For dissolved iron, the analytical uncertainty was comparable in
validation and in the subsequent quality control (2.1% and 2.5% respectively). The
sampling uncertainty was lower when sampling just one well on different occasions
during quality control (3.63.8%) than when sampling different wells at the same time
during validation (1015%). The uncertainty between-target (variation from one
sampling occasion to the next) during quality control was small for dissolved iron
(9.9%), but larger for redox potential (23%).
If a continuous control of sampling uncertainty had been required, the control data
could have been plotted in control charts in order to obtain an early warning of
excessive uncertainty (random errors) for each sampling occasion.
The number of replicates (six) in this study was less than used in most cases and the
risk of a decreased confidence in the uncertainty estimates should be considered in
evaluation of the results.
The uncertainty contribution from sampling bias was only addressed through
evaluation of the consistency of the measurements obtained from different,
interrelated chemical parameters (oxygen, dissolved iron, redox) and the evaluation
supported the conclusion that sampling and sample pre-treatment had succeeded in
avoiding bias due to oxygen impact and filter clogging.

8 Summary
The measurement uncertainty (% uncertainty with coverage factor 2) is summarised
below for dissolved iron.
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 Example A3
The data show that the requirement for less than 20% expanded measurement
uncertainty could be fulfilled for dissolved iron (sampling validation), and that the
required measurement uncertainty was in reality achieved during the routine
monitoring (sampling quality control). Furthermore, the data show that if an
improvement of the certainty of monitoring was required, the obvious point of
improvement would be increased monitoring density for dissolved iron (between-
target uncertainty dominating), whereas improvement of the on-line measurement
uncertainty could help for redox potential (large contribution of analysis uncertainty).

Dissolved iron Expanded uncertainty, coverage factor of 2 Between-target

in groundwater variability
Sampling Analysis Measurement
Validation 10% 2.1% 10% 35%l
Quality control 3.6% 2.5% 4.4% 9.9%m

9 Acknowledgements
The work presented here has been supported by Nordic Innovation Centre, the Soil
and Ground Water Contamination Committee of the Danish Council of Technical
Sciences and rhus County, Denmark. Fieldwork has skilfully been done by Mogens
Wium, GEO.

l
In the validation study, between-target variability was between wells.
m
In the quality control, between-target variability was between sampling occasions.

UfS:2007 Page 54
 Example A4

Example A4: Vitamin A in baby porridge containing fruit and milled

cereals

Measurand Uncertainty estimation

Analyte/ Unit Sector/ Sampling Purpose Design Statistics
Technique Matrix target
Vitamin A g 100 g-1 Food/ Produced Total Empirical One-way
(as in powder Baby batch measurement duplicate ANOVA
retinol)/ porridge- uncertainty method
HPLC powder
containing
fruit

1 Scope
The scope is to estimate the measurement uncertainty and contributions from
sampling and analyses. The estimates are based on samples from one type of baby
porridge taken from 10 different batches using a sampling protocol collecting
duplicate samples from each batch.

2 Scenario and sampling target

In the production of baby (infant) porridge, the vitamin A (retinol) is added as a
premix (together with vitamin D and vitamin C). The premix is a minor ingredient.
All ingredients are mixed thoroughly before distribution into packages. Earlier
analysis indicated a bigger variation in analytical result between packages than
expected. A measurement uncertainty of 2030% would be considered acceptable.
The question was raised whether the variation is due mainly to analytical uncertainty
or to sampling uncertainty. One of the theories suggests that the vitamin is locally
unevenly distributed within the package, and therefore will give bigger analytical
uncertainty if the test portion is too small (e.g. 35 g).n One possible explanation of
the heterogeneity is that the vitamin premix aggregates in small hot spots, due to
electrostatic interactions with the fruit particles in the porridge powder. The producers
recommend a test portion size of 4050 g whenever analysing vitamin A, D and C in
baby porridge powder.
In order to compare the measured vitamin A concentration against declared values
and European regulatory thresholds, an estimation of measurement uncertainty is
desirable. To determine the random component of the measurement uncertainty, an
empirical approach using the duplicate method (see Section 9.4.2) was chosen. To
estimate the systematic component a comparison with a reference value was made.

n
EN-12823-1 Foodstuffs determination of vitamin A by HPLC indicates a test sample of
approximately 210 g.
UfS:2007 Page 55
 Example A4
3 Sampling protocol
Normally a spot sampling approach is employed in which one sample (one package)
of a batch is used as a screening sample by comparing its content against the declared
values and legal limits.
Validation In this study two samples are collected from each of 10 different batches
of one type of baby porridge powder (i.e. 10 sampling targets). Each sample is equal
to one package of approximately 400 g powder.
Quality control Quality control (QC) of sampling from different types of baby
porridge is done by collecting two samples from each of eight batches of different
types of baby porridges (i.e. eight sampling targets). All the types of porridges contain
fruit in addition to milled cereals.
To ensure the quality in each package of the product at the time of the best before
date of the porridge powder, the producer wraps the product in an air-tight and light-
protecting bag. It is therefore assumed the degradation of the vitamin A is negligible
during normal shelf life. The sampling for the validation was performed at the place
of production. For QC, the samples were purchased partly at the place of production,
and partly at the retailers. When the samples were collected from retailers, care was
taken to collect the two samples (of each product) at different retailers but in addition
to assure the samples had the same batch marking. This is important to avoid adding
between-batch variations to the apparent sampling uncertainty, as the sampling
protocol in this case specifies sampling from a particular batch.

4 Study design empirical approach

An empirical (top-down) approach duplicate method was selected to provide
estimates of the random component of sampling uncertainty. The validation is
performed on one type of baby porridge containing fruit and milled cereals. In the
sampling for the QC different products of baby porridge (all containing fruit and
milled cereals) are tested to see if the estimate for measurement uncertainty from the
validation study is appropriate for different types of baby porridges containing fruit
and milled cereals.

4.1 Validation
Samples are collected on-line (just after the filling operation of packages) at random
times. Two samples (two packages, each of approximately 400 g) are collected from
each of 10 production units (batches) of one type of baby porridge powder.

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 Example A4
Figure A4.1: Sampling for validation. Two samples are taken from each of 10
production units/batches of the same type of baby porridge

Batch 1 Batch 10
S2
.. Etc. S2
S1

S1

Sample B10S2 Sample B10S1

Sample B1S2 Sample B1S1

Test sample 1 Test sample 1

Test sample 1 Test sample 1 Test sample 2 Test sample 2

Test sample 2 Test sample 2

4.2 Quality control

For quality control (QC) two samples are collected from one batch of each of eight
different types of baby porridges, containing fruit and milled cereals. The porridges
are products from three different producers. The samples (except for two types of
porridges) were provided by two of the producers. The rest was bought at the retailer.

Figure A4. 2: Sampling for QC. Two samples are taken from one batch of each
of eight different types of baby porridge

Product 1 Product 8
S2
S1
S1 .. Etc. S2

Sample P1S1
Sample P1S2 (1 package)
(1 package)

Test sample 1
Test sample 1

Test sample 2
Test sample 2

5 Sampling and analysis in the laboratory

The analytical work is done by The National Institute of Nutrition and Seafood
Research (NIFES). The laboratory is accredited according to EN ISO/IEC 17025.

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 Example A4
The laboratory participates in Laboratory Proficiency Tests (FAPAS and Bipea) with
good results (in the period 20002005, |Z-score|<1). The method was validated using
a certified reference material (CRM). Data concerning the laboratory performance is
given in Table A4.1.

Table A4.1: Methods and performance data from quality control laboratory
analyses
Parameters Vitamin A determined as retinol
Method HPLC normal phase column UV-detection
Repeatability 2RSD (%) = 6
Within-reproducibility 2RSD(%) = 8
Measurement uncertainty 14% (95% confidence interval)
Recovery Standard addition, in lab: 90110%
Based on laboratory proficiency tests (in
period 19992005), different matrixes: 88
113%, mean recovery 100.5%
Limit of quantification (LOQ) 0.14 mg kg-1
CRM used NIST 2383 baby food (mixed food composite)
CRM certified level 0.80 0.15 mg kg-1 (95% confidence interval)
CRM analysed value 0.77 0.14 mg kg-1 (n=28, 95% confidence
interval)

5.1 Secondary sampling

A mechanical sample divider (Retsch) is used to split the samples. From each of the
primary samples, four test samples are collected: two portions of approximately 35 g
and two portions of approximately 4050 g.

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 Example A4

Figure A4.3: Splitting of the primary sample to make four test samples

Batch 1 Batch 10
S2
S1
S1 .. Etc. S2

Sample B1S1
Sample B1S2
(1 package)

Test sample S2A1 Test sample S2B2

(40-50 g) (3 5 g)

Test sample S2B1 Test sample S2A2

(3-5 g) (40 50 g)

5.2 Analyses
The analytical method is based on EN-12823-1 (Foodstuffs Determination of
vitamin A by HPLC Part 1: Measurement of all-trans-retinol and 13-cis-retinol).
Retinol is saponified by using ethanolic potassium hydroxide containing antioxidants.
Vitamin A is extracted by using hexane. Analysis is performed by using high
performance liquid chromatography (HPLC), with a UV detector.
In the validation, for each of the primary samples, two analyses are performed on test
samples of 4050 g and two analyses on test samples of 35 g. In the QC two
analyses are performed on test samples of 4050 g. On each test sample only one
analytical determination is performed (no analytical duplicates).

6 Information from the producer

Data for estimating the true value of vitamin A in baby porridge are provided by the
producer (Nestl) of the product chosen for the validation, see Table A4.2.

UfS:2007 Page 59
 Example A4
Table A4.2: Product data provided by the producer
Oatmeal porridge with bananas and
Product
apricots (Nestl)
Weight of batch, including premix
1092 kg
(1 batch = 2 mixing containers)
Weight of added vitamin-premix in batch 1.228 kg
Vitamin A in premix (data from the Certificate
9016 IU g-1 = 2705 g g-1 (retinol)
of Analysis)
Vitamin A added to the batch 304 g 100 g-1 (retinol)
Vitamin A in ingredients according to the
45 g 100 g-1 (retinol)
product specification
Estimated true value of vitamin A 349 g 100 g-1 (retinol)
Retinol (sum of trans- and cis-
Vitamin A declared as
retinol)

7 Results
Test sample 40 g baby porridge

Table A4.3: Validation data from the same product, results given in g 100 g-1
powder
Batch S1A1 S1A2 S2A1 S2A2
B1 402 325 361 351
B2 382 319 349 362
B3 332 291 397 348
B4 280 278 358 321
B5 370 409 378 460
B6 344 318 381 392
B7 297 333 341 315
B8 336 320 292 306
B9 372 353 332 337
B10 407 361 322 382

S1 and S2: Primary samples from sampling location 1 and 2 of one production batch
A1 and A2: Analyses of duplicate test samples of a primary sample S
Analysed mean value (test sample 40 g): 348 g 100 g-1

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 Example A4
Test sample 4 g baby porridge

Table A4.4: Validation data same product, results given in g 100 g-1 powder
Batch S1B1 S1B2 S2B1 S2B2
B1 400 491 323 355
B2 413 159 392 434
B3 315 391 252 454
B4 223 220 357 469
B5 462 343 262 293
B6 353 265 305 456
B7 298 234 152 323
B8 425 263 417 353
B9 622 189 291 272
B10 292 397 142 568

S1 and S2: Primary samples from sampling location 1 and 2 of one production batch
B1 and B2: Analyses of duplicate test samples of a primary sample S
Analysed mean value (test sample 4 g): 341 g 100 g-1

7.1 Calculations
The ANOVA calculation can be done by using available tools in Excel, Minitab,
SPSS etc. In this study the calculations are done in an excel spreadsheet and the
details are shown in Section 10 ANOVA calculations.
Calculation of uncertainty of analyses, one-way ANOVA, test sample 40 g

Table A4.5: Results from ANOVA calculations uncertainty of analyses sum

of squares of differences, within-groups (SS-Error). For details see Section A4.11
Standard deviation,
SSE-Anal Degrees of Variance SDanal Relative standard deviation
freedom (df) RSDanal(%)
(g 100 g-1)2 = SSE-Anal /df
= (SSEAnal) /df
(g 100 g-1)2 = (SD / X a)*100%
(g 100 g-1)
16595 20 829.75 28.805 8.28

Calculation of uncertainty of sampling, one-way ANOVA, test sample 4 g

Table A4.6: Results from ANOVA calculations uncertainty of sampling sum

of squares of differences. For details see Section A4.11
Variance
SSS Degrees of Standard deviation, Relative standard
-1 2 freedom (df) VSamp= SDsamp deviation RSDsamp(%)
(g 100 g )
(SSS/dfS SSEAnal/dfA)/2 = V Samp (g 100 g-1) = (SD / X s)*100%
(g 100 g-1)2
14231 10 296.7 17.22 4.95

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 Example A4
Calculation of measurement uncertainty 40 g test sample
The RSD (%) value from the ANOVA calculation can be used as an estimate of the
standard uncertainty u (%). The analytical laboratory has estimated the analytical
standard uncertainty to be 7%, which is lower than the random analytical component
for this sample type of 8.28%. The higher value of these two is used in the
calculations. Combining the RSD values from tables A4.5 and A4.6 with Equation 1,
the results can be written as in Table A4.7.

umeas = (u samp ) 2 + (u anal ) 2 (Equation A1)

Table A4.7: Measurement uncertainty 40 g test sample

Measurement uncertainty, ANOVA calculations 40 g test samples
Sampling Analytical Total
Uncertainty u (%) 4.95 8.28 9.7
Expanded uncertainty U (%) = 2*u
With a coverage factor of 2 (i.e. 95% 9.9 17 20
confidence)

Calculation of uncertainty of analyses, one-way ANOVA, test sample 4g

The same calculations are used as for the test sample size of 40 g (see Table A4.14, in
Section 11 of this example).

Table A4.8: Results from ANOVA calculations uncertainty of analyses, 4 g test

sample sum of squares of differences, within groups (SS-Error)
Standard deviation,
Degrees of Variance SDanal Relative standard deviation
SSE RSDanal(%)
freedom (df) = SSE /df
(g 100 g-1)2 = SSE /df
(N*2-N)=20 (g 100 g-1)2 = (SD / X a)*100%
(g 100 g-1)

312206.5 20 15610.325 124.9413 36.6800

Calculation of uncertainty of sampling, one-way ANOVA, test sample 4 g

Table A4.9: Results from ANOVA calculations uncertainty of sampling, 4 g test

sample sum of squares of differences
Variance Standard deviation,
SSS Degrees of SDsamp Relative standard deviation
VSamp=
-1 2 freedom (df) RSDsamp(%)
(g 100 g )
(SSS/dfS = V Samp
SSEAnal/dfA)/2 = (SD / X s)*100%
(g 100 g-1)2 (g 100 g-1)

- 2662.15
102860.25 10 -2662.15 Conventionally set to zero
Set to zero

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 Example A4
The same calculations are used as for the test sample size of 40 g (Table A4.15,
Section 11 of this example).
The negative value of VSamp in Table A4.9 indicates that SDSamp is small compared to
the calculated value of SDanal. In this case, the estimates of SDanal and SDsamp using
robust ANOVA confirmed the smaller sampling standard deviation; the robust
ANOVA estimates were uSamp(%) = 6.9% and uAnal(%) = 30%.
As the sampling is identical for the experiments with 40 g and 4 g test samples (and
the uncertainty of sampling therefore should be the same), an RSDsamp (%) = 5%
( 4.95 see table A4.7) is used as an estimate.
Calculation of measurement uncertainty 4 g test sample
Using the calculated RSD (%) value in Tables A4.5 and A4.6 as an estimate of the
measurement uncertainty and combining with Equation A1 the results can be written
as follows (Table A4.10).

Table A4.10: Measurement uncertainty 4 g test sample

Measurement uncertainty, ANOVA calculations 4 g test samples
*Sampling Analytical Measurement
Uncertainty u (%) 5 36.7 37
Expanded uncertainty U (%) = 2*u
With a coverage factor of 2 (i.e. 95% 10 73.4 74
confidence)

* The u (%) value is derived from calculations using 40 g test samples

7.2 Effect of the size of test sample on measurement uncertainty

The baby porridge powder looks homogeneous, and therefore a low measurement
uncertainty (u) is expected. However, analyses of the powder indicated a surprisingly
large u when using a test sample size of 4 g (the CEN-standard EN-12823-1 indicates
a test sample size of approximately 210 g). The producers recommended using a test
sample size of 4050 g.
The validation tests gave the following results

Table A4.11: Comparing measurement uncertainty test samples of 40 g and 4 g

Test sample size Measurement Expanded measurements
uncertainty (umeas) uncertainty Umeas
40 g test sample 9.7% 20%
4 g test sample 37% 74%

It can be concluded that u40g << u4g. A Umeas of approximately 20% is acceptable,
using the manufacturers criterion of 2030%, while a Umeas of 74% is considered to
be too high, taking into account the matrix and production conditions of this type of
product.

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 Example A4
It can therefore be concluded that a test sample weight of 4 g is not fit for purpose
when analysing vitamin A (retinol) in baby porridge powder containing milled cereals
and fruit. A test sample size of 4050 g is recommended. This also supports the theory
that the vitamin is unevenly distributed in the product, possible as local hot spots
due to electrostatic interactions.

7.3 Quality control

According to Section 13.2.2 of this Guide, the principal tool in quality control is
replication. This is minimally executed by taking two samples from each target by a
complete (and suitably randomised) duplication of the sampling protocol. There is
only a need to analyse the sample once and the difference between the results
D = x1 x2 is calculated. In this study each sample was analysed twice, but the
comparisons were made between one analyses of each sample (double set).
In the quality control study, test portions of 40 g were used. According to
declarations, the products contain different amounts of vitamin A.

Table A4.12: Quality control data for test portion 40 g of different products
Product Producer Porridge powder ingredients S1A1 S1A2 S2A1 S2A2
P1 1 Oat, rice and pear 322 319 350 375
P2 1 Oat, rye, rice and pear 332 317 358 393
P3 1 Wheat, banana and apple 443 430 461 388
P4 1 Wheat and apple 318 383 390 334
P5 2 Oat, rice and banana 252 219 265 227
P6 2 Wheat and apple 274 239 233 217
P7 2 Oat, rice and apple 206 225 198 195
P8 3 Wheat, spelt, oat and apple (organic product) 392 335 375 416

S1 and S2: Primary samples (laboratory samples) from sampling locations 1 and 2 of
one batch from each product.
A1 and A2: Analyses on two test samples from each laboratory sample.

Quality control calculation and control chart

The validated uncertainties of sampling and analysis are usamp and uanal respectively.
The construction of a control chart is described in Section 13.2. In the case of baby
porridge (40 g test sample) the following calculations can be made:

Warning limit: WL = 2.38 * u 2 anal + u 2 samp = 2.83 * (4.95 2

+ 8.282 )% = 27%

Action limit: AL = 3.69 * (4.95 2

+ 8.28 2 )% = 36%

Central line: CL = 1.128 * (4.95 2

+ 8.28 2 )% = 11%

UfS:2007 Page 64
 Example A4
Table A4.13: Quality control: calculation of differences D and D (%) between
samples from a batch

Sample S1 Sample S2 D(%) = ( D / x) * 100%

D = xS1 xS 2 x
Product Analyses XS1 XS2
P1 A1 322 350 28 336 8
P2 332 358 26 345 8
P3 443 461 18 452 4
P4 318 390 72 354 20
P5 252 265 13 259 5
P6 274 233 41 254 16
P7 206 198 8 202 4
P8 392 375 17 384 4
P1 A2 319 375 56 347 16
P2 317 393 76 355 21
P3 430 388 42 409 10
P4 383 334 49 359 14
P5 219 227 8 223 4
P6 239 217 22 228 10
P7 225 195 30 210 14
P8 335 416 81 376 22

The calculated D (%) in Table A4.13 can be compared directly with the action limit,
or the results can be presented in a control chart, see Figure A4.4.

UfS:2007 Page 65
 Example A4

Figure A4.4: Control chart: quality control analyses of vitamin A in baby

porridge containing cereals and fruits

40
Difference between results - d(%)

30

Calculated: D (%)
Action limit (36 %)
20
Warning limit (27 %)
Central line (11%)

10

0
P1-A1

P2-A1

P3-A1

P4-A1

P5-A1

P6-A1

P7-A1

P8-A1

P1-A2

P2-A2

P3-A2

P4-A2

P5-A2

P6-A2

P7-A2

P8-A2
Product/analyses

The control chart in Figure A4.4 shows that when collecting duplicated samples from
the same batch, the difference between analytical results D (%) is smaller than the
action limit. All the calculated difference are in fact smaller than the warning limit of
27%.
The measurement uncertainty determined in the validation step is therefore considered
suitable for the quality control of the sampling of baby porridge containing milled
cereals and fruit.
If the normal procedure is to analyse one sample from each batch, it is recommended
that duplicate samples are collected from the same batch at least in one of ten of the
sampled batches.
Measurement uncertainty
Sampling uncertainty
Calculations from the validation study gave an expanded sampling uncertainty Usamp
(%) = 10% (40 g test sample see Table A4.7). The calculated uncertainty does not
include contributions to the uncertainty due to between protocol and between
samplers differences.
Analytical uncertainty
Calculation from the validation study gave an expanded measurement uncertainty of
analyses (Uanal) of 17% 40 g test sample. The laboratory reports their own
estimation of the analytical uncertainty (see Table A4.1): 2*RSDinlab(%) = 14%.
2*RSDinlab(%) is used as an estimate of Uanal in the laboratory. The Uanal found in the
validation study was at the same level but still a little bigger than the Uanal reported by
the laboratory.
The certified reference material (CRM) used is 2383 (NIST) baby food composite.
The CRM is a mix of different foods of plant and animal origins and the uncertainty
found when analysing the CRM might not be identical with that found when analysing
baby porridge powder. Laboratory data for the CRM 2383 is included in the table
below.
UfS:2007 Page 66
 Example A4

Mean value mg Laboratory

CRM 2383 kg-1 U (%)95% bias (%)
Certified 0.80 0.15 18.8 -
Analysed 0.77 0.14 18.2 - 3.75

The measurement uncertainty and the bias determined for the CRM could be allowed
for in the analytical measurement uncertainty (as in the NordTest UFS Guide,
Example 2), but as the matrix in the validation study is different from that for the
CRM used, we chose not to include it in this study.
Total measurement uncertainty
Calculations from the validation study gave an expanded measurement uncertainty
Umeas(%) = 20% (40 g test sample see Table A4.7).
Systematic bias
The laboratory reports a recovery of normally 90110%. Recovery based on
laboratory proficiency tests 19992005: 88113%. The results for the PT indicate no
(or very small) systematic bias. Analyses of CRM 2383 in the laboratory gives a mean
analysed value of 96.3% of the certified value which indicates a small bias (-3.7%).
As the matrix of the CRM baby food composite is different to the baby porridge,
and the analytical method includes an extraction, the bias determined when analysing
the CRM might not be representative for the analyses of baby porridge.
In the validation study, the mean value of retinol was determined to be 348 g 100 g-1
(when using a test sample of 40 g). According to data provided by the producer (see
Table A4.2), the true value for retinol was calculated to be 349 g 100 g-1 porridge
powder. This gives a recovery of 99.7% of the true value. This indicates that the
systematic error due to sampling and analyses is small and might be negligible when
analysing baby porridge-powder containing milled cereals and fruits on the
condition that a test sample of 4050 g is used.

8 Comments
When a test sample of approximately 40 g is used, the retinol concentration C in baby
porridge-powder containing milled cereals and fruit should be reported with the
expanded measurement uncertainty, i.e. C20% of the measured value C (95%
confidence).
When baby porridge-powder containing milled cereals and fruit is to be analysed, it is
recommended to use a relatively large test sample of approximately 4050 g and not
210 g as indicated in the official CEN method (EN-12823-1). As the analytical
uncertainty (40 g test sample) was bigger than the normal analytical uncertainty of the
laboratory, even larger samples than 40 g might be considered.

UfS:2007 Page 67
 Example A4
9 Summary
Measurement uncertainty for 40 g test samples Sample
Sampling Analytical Total Typical between-target variation
RSDB(%) of the mean values of
analyses of the batches in the
validation test (see Table A4.15)
Uncertainty u (%) = 4.95 8.3 9.7 8.2
RSD (%)
#
Expanded uncertainty 9.9 16.6 19.4 16.4
U (%) = 2*u
#
With a coverage factor of 2 (i.e. 95% confidence)

10 Acknowledgements
Nestl (Norway) is thanked for its enthusiastic cooperation and in addition for
providing samples to the project (validation and quality control study). Also Smaafolk
Tine Norske Meierier is thanked for kindly offering us samples for the quality
control study. The National Institute of Nutrition and Seafood Research (NIFES) is
thanked for the analytical contribution (analyses and information on the laboratory
QA-system). The study was done with financial support from the Nordic Innovation
Centre and the Norwegian Food Safety Authority.

UfS:2007 Page 68
 Example A4

11 ANOVA calculation, vitamin A in baby porridge details

Calculation of uncertainty of analyses, one-way ANOVA, test sample 40 g

Table A4.14: ANOVA calculations uncertainty of analyses sum of squares of

differences, within groups (SS-Error)
Mean value each
Squares of differences
-1
Analyses (g 100 g ) sample
within groups (g 100 g-1)2
(g 100 g-1)

Sample A1 =xi j =xi1 A2 = xi j = xi2 2

xi = ( xi1+ xi2)/2 (xi- x i)
B1-S1 402 325 363.5 1482.25
B2-S1 382 319 350.5 992.25
B3-S1 332 291 311.5 420.25
B4-S1 280 278 279 1
B5-S1 370 409 389.5 380.25
B6-S1 344 318 331 169
B7-S1 297 333 315 324
B8-S1 336 320 328 64
B9-S1 372 353 362.5 90.25
B10-S1 407 361 384 529
B1-S2 361 351 356 25
B2-S2 349 362 355.5 42.25
B3-S2 397 348 372.5 600.25
B4-S2 358 321 339.5 342.25
B5-S2 378 460 419 1681
B6-S2 381 392 386.5 30.25
B7-S2 341 315 328 169
B8-S2 292 306 299 49
B9-S2 332 337 334.5 6.25
B10-S2 322 382 352 900
Mean value of measurements: 2
SS-Error (SSE):
20 20 20
X a =1/20 *
i =1
x i = 347.85 g 100 g-1 = [(xi1- x i)2+(xi2- x i)2]= 2* (xi- x i)2
i =1 i =1

Standard deviation,
Degrees of Variance SDanal Relative standard deviation
SSE RSDanal(%)
freedom (df) = SSE /df
(g 100 g-1)2 = SSE /df
(N*2-N)=20 (g 100 g-1)2 = (SD / X a)*100%
(g 100 g-1)
16595 20 829.75 28.80538 8.280978
Notes on Table A4.14.
1. Calculation of SS-Error in this case two test samples are analysed for each laboratory sample, therefore:

(xi1 xi )2 = (xi 2 xi )2 SSE = [(xi1 xi )2 + (xi 2 xi )2 ] = 2 (xi1 xi )2

20 20

i =1 i =1

UfS:2007 Page 69
 Example A4
If the number of test samples analysed is greater than two, the squares of differences will be not be equal and the
20

(x xi )
n
2
calculation to be done is the following: SSE = ij
i =1 j=1

2. df = (N*n-N)=(20*2-20)= 20 where N is the number of samples and n is the number of test samples analysed of
each batch.

Calculation of uncertainty of sampling, one-way ANOVA, test sample 40 g

Table A4.15: ANOVA calculations uncertainty of sampling sum of squares of

differences
2
x
i xi1 + xi 2 xi 3 + xi 4
2
S1A1=x i1 S1A2=xi2 S2A1=xi3 S2A2=xi4 xi xi xi
2 2
402 325 361 351 359.75 14.0625 14.0625
382 319 349 362 353 6.25 6.25
332 291 397 348 342 930.25 930.25
280 278 358 321 309.25 915.0625 915.0625
370 409 378 460 404.25 217.5625 217.5625
344 318 381 392 358.75 770.0625 770.0625
297 333 341 315 321.5 42.25 42.25
336 320 292 306 313.5 210.25 210.25
372 353 332 337 348.5 196 196
407 361 322 382 368 256 256

10

2 2 2 2
x + xi 2 x + xi 2 x + xi 4 x + xi 4
SS Samp = i1 x i + i1 xi + i 3 xi + i 3 xi

i =1 2 2 2 2
10

2 2
x + xi 2 x + xi 4
= 2 * i1 xi + 2 * i 3 x i = 14231

i =1 2 2

Mean value of all measurements x = 347.85 RSDSamp(%)=(SDSamp/ x )*100%= 4.95%

dfS =10 (see table note)
SSEAnal = 16595 (see Table A4.14)
dfA = 20 (see Table A4.14)
Variance VSamp= (SSS/dfS SSA/dfA)/2
SDSamp = Vsamp = 17.224
= (14231/10 16595/20)/2 = 296.675

Notes on Table A4.15.

x + xi 2 x i 3 + xi 4
1. The difference d between the mean value x of the two values i1 and to each of
2 2
the values are identical. The expression could therefore be written as
10 2 10 xi1 + xi 2
2

SS Samp = 4 * d i = 4 * xi
i =1
i =1 2
2. dfs= (NB*n-NB)=(10*2-10)= 10 where NB is the number of batches and n is the number of primary samples (=
laboratory samples) analysed for each batch.

UfS:2007 Page 70
 Example A5
Example A5: Enzyme in chicken feed

Measurand Uncertainty estimation

Analyte/ Unit1 Sector/ Sampling Purpose Design Statistics
Technique Matrix target
Enzyme/ % m/m Food & 25 kg bag Total Modelling Summation
HPLC (i.e. mass Feed/ uncertainty with of
fraction) Chicken feed (weak links sampling component
in theory (of variances
measurement Gy)
chain)
1
Including reporting base

1 Scope
The scope is to estimate the sampling uncertainty with the given sampling protocol by
applying Gys sampling theory (Section 10.2). The analyte is an added enzyme
ingredient in the feed. Sampling theory provides realistic estimates only if all
sampling and sample splitting operations are carried out obeying the rules of sampling
correctness; it is assumed in this example that no gross errors are present and that
incorrect sampling errors are negligible.

2 Scenario and sampling target

An enzyme product is used as an additive in chicken feed (density = 0.67 g cm-3). The
nominal concentration of the enzyme is 0.05% m/m. The enzyme powder has a
density of 1.08 g cm-3. Powders are carefully mixed. The size distribution of the
enzyme particles was known and it was estimated that the characteristic particle size
was d = 1.00 mm and the size factor was g = 0.5. The purpose of this exercise is to
estimate the total uncertainty of the protocol (i.e. as fundamental sampling error,
Section 10.2.7 and Figure 4) used for estimating the average content in each 25 kg bag
employed to ship the product to customers.

3 Study design, using a modelling approach (bottom-up)

A model is constructed using sampling theory as described in Section 10.2. The
parameters are either measured directly, or estimated, and assumed to be single values
and to be constant within and between each bag.

4 Sampling and analysis in the laboratory

The actual concentration of the enzyme in the sampling target, which is identified as a
25 kg bag, is estimated by taking a 500 g primary sample from it.
The material from the primary sample is ground to a particle size of <0.5 mm. Then
the enzyme is extracted from a 2 g test portion by using a suitable solvent and the
concentration is determined by using liquid chromatography. The relative standard
deviation of the chromatographic measurement, estimated from the laboratory quality
control data, is 5%.
UfS:2007 Page 71
 Example A5
5 Results
To estimate the fundamental sampling error (FSE, Section 10.2.7, Figure 4) of the two
sampling steps, we have the to evaluate the material properties (Table A5.1).

Table A5.1: Input values for estimation of sampling uncertainty by the modelling
approach, using sampling theory
Primary Secondary sample Comment
sample
M1 = 500 g M2 = 2.0 g Sample sizes
ML1= 25,000 ML2 = 500 g Lot (sampling target) sizes
g
d1 = 0.1 cm d2 = 0.05 cm Particle sizes
g1 = 0.5 g2 = 0.25 Estimated size distribution factors
Both samples
aL = 0.05% m/m Mean concentration of enzyme in the lot
= 100% m/m Enzyme concentration in enzyme particles
c = 1.08 g cm-3 Density of enzyme particles
m = 0.67 g cm-3 Density of matrix particles
f = 0.5 Default shape factor for spheroidal
particles
=1 Liberation factor for liberated particles

These material properties give for the constitution factor (Equation 7) the value c =
2160 g cm-3 and for the sampling constants (Equation 6) C values
C1 = 540 g cm-3 and C2 = 270 g cm-3
Equation 5 can be used to give estimates of the standard deviation for each sampling
step (as estimates of the standard uncertainty).
sr1 = 0.033 = 3.3% . Primary sample
sr2 = 0.13 = 13% . Secondary sample
sr3 = 0.05 = 5% . Analytical determination
The total relative standard deviation (st, combined uncertainty) can now be estimated
by applying the rule of propagation of errors; for i errors we have:

st = s 2
ri = 0.143 = 14.3 %

The relative expanded uncertainty, with a coverage factor of 2, is therefore 28.6%

(excluding analytical uncertainties associated with systematic effects, such as
analytical bias).

6 Comments
The largest source of uncertainty in the whole measurement process is identified as
that generated in preparing the test portion (2 g) for the extraction of the enzyme.
No additional allowance has been made for uncertainties associated with systematic
effects during analysis, and incorrect sampling errors (and sampling bias) have been
assumed to be negligible.
UfS:2007 Page 72
 Example A5
7 Assessment of fitness for purpose of these measurements
If it is decided that the overall uncertainty of 28.6% is not fit for purpose (Section 16),
then it is the step in which the test portion is prepared that needs to be modified, to
reduce the overall uncertainty. Either a larger sample should be used for the
extraction, or the primary sample should be pulverised to a finer particle size,
whichever is more economic in practice. The model can also be used to predict either
the increase in mass, or reduction in particle size, that is required to achieve the
uncertainty that will be considered fit for purpose (e.g. Appendix E).

8 Reporting and interpretation

Measurement of the enzyme concentration reported for each 25 kg bag should have an
attached uncertainty of 28.6% of the concentration value. The continued use of this
uncertainty value will depend on the periodic checking of the validity of the values
and assumptions used in its calculation.

9 Summary

Measurement uncertainty*
Sampling Analytical Total
26.8% (rel) 10.0% (rel) 28.6% (rel)
* with coverage factor of 2 (i.e. 95% confidence)

UfS:2007 Page 73
 Example A6

Example A6: Cadmium and phosphorous in agricultural top soil by

modelling approach

Measurand Uncertainty estimation

Analyte/ Unit Sector/ Sampling Purpose Design Statistics
Technique Matrix target
Cd: GF-ZAAS mg kg-1 Environ- Arable soil Total Modelling Summation
direct solid mental/ uncertainty approach of
to air dried 143 x 22 m,
sampling agricultural (with (using component
basis depth 30 cm
top soil contributions exploratory variances
from each measure-
P: Ca-Acetate sampling ments for
Lactate (CAL) effect) single
method effects)

1 Scope
Estimation of the overall uncertainty of measurement by summation of individual
uncertainty contributions from sampling, sample preparation and analysis using the
modelling approach.

2 Scenario and sampling target

Investigation aims to estimate the mean concentration of cadmium and phosphorus in
top soil of a target that is an area of arable land of 0.32 hectare (specification of the
measurand). Sampling used composite samples in a protocol that is commonly applied
to agricultural control.

3 Sampling protocol
The target area was sampled using a stratified protocol, with a sampling density of
approximately 20 increments per hectare, to a depth of 30 cm, using a soil auger.

4 Study design cause-and-effect modelling approach (Section 10.1)

4.1 Identification of effects in the measurement

The following sources can be considered as potential significant contributors to the
uncertainty in the general case.
4.1.1 Sampling
The spatial distribution of the analyte over a two-dimensional object creates two
different uncertainty components long range point selection error (Appendix C2.3):
The sampling variance of the analyte content between composite samples from
different locations characterises the statistical distribution of the analyte over the
target area. This value often depends on the distance between sampling
points/sampling locations.

UfS:2007 Page 74
 Example A6
If the spatial pattern of the analyte on the area is not represented by the sampling
pattern (sampling strategy), sampling bias may occur.
With the use of a sampling tool, different effects may appear, such as point
materialisation error (Figure 3). This may occur due to an ill-defined reference level
of the soil (e.g. due to undulation of the soil surface or difficulties in the definition of
the horizons), or variation in the actual sample depth or in soil density (e.g. by
moisture content), or by selective loss of soil material from the sampling device.
These effects only lead to an uncertainty contribution, if there is a depth gradient in
the analyte content (a third dimension to the target body). For this reason, these
effects, which are difficult to determine one by one, are summarised collectively as
the depth-effect.
4.1.2 Sample preparation
The physical sample preparation comprises the step from the field sample to the
laboratory sample. Mechanical treatment, such as disaggregation, sieving, grinding
and splitting steps, reduce the amount of soil material. With these steps errors may
arise due to variation in duration and forces of mechanical treatment, heterogeneity,
segregation of different soil (particle) fractions and particles size distribution. A
periodic point selection error (Figure 3) may occur due to variation in moisture
content of the dried soil sample by sorption/desorption of water from air to an
equilibrium state (depending on the humidity and properties of the sample material,
e.g. particle size).
4.1.3 Analysis
The analysis is the third step of the measurement process, which is connected with
different kinds of effects that give rise to uncertainty contributions. The analytical
uncertainty of the laboratory samples can be estimated by previously published
procedures [1, 35]. The separation of the laboratory sample into analytical test
samples will add to the sampling uncertainty; specifically, another fundamental error
may occur. However, the random component of this sampling effect is included in the
analytical repeatability precision between test samples. A significant systematic
component should be avoided by proper mixing of the sampling powder.

4.2 Cause-and-effect diagram

Figure A6.1 shows the cause-and-effect diagram for the measurement process. In the
sampling and sample preparation steps the sources of uncertainty contributions are
given; for the analysis, only the analytical quality parameters are indicated.

4.3 Model equation

The input quantities of the sampling effects discussed above are not constituent parts
of the equation from which the measurement result is calculated. An appropriate
model equation for the overall measurement process can be established, however, by
introducing respective nominal correction factors on the analytical result:
x site = x anly f b-loc f strat f depth f prep f dry

where
xsite = measurement result
x analy = mean from the analysis of test samples

UfS:2007 Page 75
 Example A6
fb-loc = correction factor for deviation between locations
fstrat = correction factor for bias due to sampling strategy
fdepth = correction factor for the depth effect
fprep = correction factor for errors during mechanical sample preparation
fdry = correction factor for deviation of moisture content

Figure A6.1: Cause-and-effect diagram for soil sampling on arable land (Rw is
within-laboratory reproducibility)
Long range Point
point selection materialisation

spatial analyte pattern 0 - level

depth
sampling pattern loss of material
number of increments moisture content

sampling strategy "depth effect"

sample preparation
RW
xsite
heterogeneity
selective loss
bias
mech. force
drying
humidity
C ref
temperature
material properties
Mechanical sample Analysi s
preparation

If no significant bias is detected, all correction factors can be set to unity so that the
best estimate for the measurand is given by:
x site = x anly
Because of the simplicity of the model equation (only factors), and assuming
independence between the factors, the combined uncertainty can be achieved by
variance addition of the relative standard uncertainties from the various effects:
u site = 2
u anly + u 2b-loc + u strat
2
+ u depth
2
+ u 2prep + u dry
2

5 Sampling and analysis in the laboratory

The sample mass was reduced by cone and quartering, air dried and sieved to select
the grain size <2 mm.
The analysis was performed by the following methods for cadmium using Zeeman-
GF-AAS (direct solid sampling) and for phosphorus using the Calcium-Acetate-
Lactate (CAL) method (the analytical measurement procedures are described
elsewhere in separate protocols).

UfS:2007 Page 76
 Example A6
6 Results of evaluation of individual effects in this case study
The estimation of the standard uncertainty from the analyte distribution over the target
area is based on a modified increment sampling based on the sampling protocol. For
elucidation of the outcome of single effects additional exploratory measurements have
been carried out.

6.1 Variation between locations

The area was divided into nine squares (A, B, C 1, 2, 3), and three increments are
taken from each of five squares (crosswise over the area). The increments from each
square are combined, resulting in five separate composite samples. These samples are
treated and analysed separately. The mean of the single results constitutes the
measurement result in agreement with the specification of the measurand.
The analytical results for both analytes under investigation are shown in Table A6.1.
The standard deviation between these values (ssqr) reflects the variation between the
composite samples for each nominate square.
The standard uncertainty in the overall mean value (i.e. the measurement result) due
to this effect can be estimated by considering the number of samples between
locations using the standard error on the mean:
s sqr
u b-loc =
n b-loc

Table A6.1: Measured concentration of cadmium and phosphorus in five squares

Cd P
Square mg kg-1 mg kg-1
A1 0.270 124
A3 0.285 112
B2 0.343 120
C1 0.355 118
C3 0.343 105
x sqr 0.319 116
s sqr 0.039 8.0
(12%) (6.5%)
ub-loc 5.4% 2.9%
The table shows the mean value across the five squares (the measurement result), the standard
deviation calculated from these values (ssqr), and the estimated uncertainty contribution from
the standard error on the mean (ub-loc).

6.2 Sampling strategy

Inspection of the analyte contents between the squares (Table A6.1) shows no notable
differences for phosphorus in any direction (neither vertical, nor horizontal, nor
diagonal). So, no significant bias (e.g. 0.5% ) in the measurement result can be
expected for this analyte from this source.

UfS:2007 Page 77
 Example A6
For cadmium both A squares show a considerably lower analyte content than the B
and C squares. Such a gradient was not unexpected for this particular area because the
C squares lay on a forest boundary, while the A squares border on grassland and the 1
and 3 squares lay between other arable land areas. It is well known that in the upper
horizon of forest soils accumulation of heavy metal occurs, which can influence
adjacent areas.
A hypothesis-based sampling pattern was applied to look for such an effect.
However, the values measured with this sampling strategy only detected a minor
systematic effect. A standard uncertainty of 1% is therefore inserted into the
uncertainty budget for cadmium for sampling strategy.

6.3 Depth effect

For revealing the collection of effects referred to as the depth effect, the following
exploratory experiment was performed.
Increment cores are taken for a depth of 35 cm within the five test squares. From
these cores segments of 2530 cm and of 3035 cm are separated and combined.
Table A6.2 shows the analytical results for these samples.

Table A6.2: Depth experiments

Cd P
mg kg-1 mg kg-1
c- (2530 cm) 0.14 47
c+ (3035 cm) 0.10 35
x- 0.34 124
x+ 0.30 109
x 0.04 15
udepth 3.5% 3.7%
The table shows the average content of the depth horizons from five cores from different
locations, the calculated content limits and the estimated uncertainty contribution
Both analytes show a statistically significant negative gradient with respect to depth.
The uncertainty due to the depth effect was estimated by considering the analyte
content of the soil layers below and above the reference depth (c-, c+) by the following
model.
The maximum variation in the sampling depth is assumed to be not more than 10%
(i.e. 2733 cm). From these data the lower and upper content limits (x -, x+), related to
the mean content of an auger core of nominal depth, are estimated according to:
x - 0.1 c _ x + 0.1 c +
x_= x+ =
0.9 1.1
The difference between x- and x+ (xdepth) is assumed to be the maximum deviation
from the mean content due to depth variation of the increments.
If a rectangular distribution for the deviation in depth is assumed, the standard
uncertainty in the mean value (Table A6.2) can be estimated by:
x depth / 2
u depth =
3
UfS:2007 Page 78
 Example A6
6.4 Splitting
The primary field samples were split in half, seven times by a coning and quartering
procedure resulting in a laboratory sample that was 1/64 of the original mass.
To reveal the splitting effect the following exploratory experiment was performed.
In the first splitting step the second half of the material was not discarded, but
considered as a duplicate sample, which was treated like the original sample and
analysed separately. Table A6.3 shows the relative standard deviations between the
duplicates of each of the five squares for both analytes.
As a simple approximation, the mean of the relative standard deviations is taken as the
standard uncertainty of the splitting step
u split = ssplit

Note: The observed large spread of standard deviations between the duplicates must
be expected. The 2-distribution for df = 1 shows high probability for very low values
and a moderate probability for large values.

Table A6.3: Relative standard deviations between duplicate split samples and the
mean of these standard deviations for both analytes
Cd P
Square (%) (%)
A1 0.44 1.49
A3 9.17 2.80
B2 5.32 0.84
C1 3.24 8.88
C3 0.44 1.81
ssplit 3.7 3.3

6.5 Drying
For the drying effect no experiment was performed, but information from the
literature was used to estimate the effect. A moisture content between 1 and 3% has
been found for a large number of air-dried soil samples [44]. According to the
sampling protocol, the measurand refers to air-dried soil material. Consequently, no
correction for moisture content is required for the concentration measurements.
However, a range of xdry = 2% difference in moisture content must be considered.
Assuming a rectangular distribution across this range, the standard uncertainty for
both analytes can be estimated as:
x dry / 2
u dry = = 0 .6 %
3

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 Example A6
6.6 Analysis
The uncertainty from the analytical process for cadmium and phosphorus (Tables
A6.4 and A6.5) were estimated from quality control data, using the Nordtest-approach
[25].

Table A6.4: Standard uncertainty components and combined uncertainty in the

analysis of the soil sample for cadmium

RW Uncertainty from within-laboratory reproducibility, uRw = 3.6%

evaluated from the repeatability standard deviation of
the mean from n=10 test samples and the instrument
stability over the working session of one day
cref Uncertainty of the certified value of a CRM uref = 2.7%
bias No uncertainty contribution from laboratory bias, -
because the results are corrected for the day-to-day bias
of the CRM measurements

sbias Uncertainty contribution from the standard deviation of ubias = 2.7%

the mean (n=3) from the day-to-day analysis of the
CRM
Combined analytical uncertainty uanly = 5.2%

Table A6.5: Standard uncertainty components and combined uncertainty in the

analysis of the soil sample for phosphorus

RW Uncertainty from within-laboratory reproducibility, uRw= 1.7%

evaluated from the repeatability standard deviation of
the mean from n=1 test samples
cref Uncertainty for the trueness of the results estimated as ubias = 9.5%
bias the reproducibility precision sR from one inter-
laboratory comparison (worse case estimate)
sbias
Combined analytical uncertainty uanly = 9.7%

6.7 Uncertainty budget and measurement result

Table A6.6 lists the evaluated standard uncertainty from the effects under
consideration. The combined uncertainty is calculated from these contributions.

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 Example A6

Table A6.6: Relative standard uncertainties from the considered effects and the
combined uncertainty for both analytes
Relative standard
uncertainty (%)
Effect
Cd P
Variation between locations 5.4 2.9
Sampling strategy 1.0 0.5
Depth 3.5 3.7
Splitting 3.7 3.3
Drying 0.6 0.6
Analysis 5.2 9.7
Combined uncertainty 9.1 11.3

Measurement result:
Cd: 0.32 0.06 mg kg-1
P: 116 26 mg kg-1
(coverage factor of 2 for approx. 95% confidence level)

7 Comments

7.1 Contribution of effects

Table A6.6 shows that the sampling/sample preparation process contributes
considerably to the overall measurement uncertainty. To recognise and to assess the
relevance of single effects/process steps several aspects must be considered:
7.1.1 The between-location effect depends on the homogeneity of the target area and
the total number of increments taken from each square. Former investigations show
that 20 increments per hectare of arable land yield an uncertainty contribution in the
order of the analytical uncertainty.
7.1.2 The error due to the sampling strategy is difficult to quantify, but can often be
much larger than that observed in this case study. Practically it can only be controlled
by expert judgement of the large-scale distribution of the analyte over the area and
the choice of an appropriate sampling strategy.
7.1.3 With the model calculation of the depth effect, it is treated as an unknown
systematic error, that is, the deviation in depth occurs with all increments (more or
less) in the same direction. This may be realistic under specific conditions; for
example, a dry sandy soil tends to drop out at the lower end of the auger so that the
average increment depth would be too small. If such an effect is detected, then the
correction of the systematic deviation is possible and only the random error
component must be considered (i.e. the uncertainty decreases with the factor of
1/nincr). Training of the sampler may reduce this point materialisation error.
7.1.4 The splitting effect is hard to control because initial mass reduction is often
performed in the field. It can contribute significantly if the method of mass reduction

UfS:2007 Page 81
 Example A6
is inappropriate or performed carelessly. Consequently, training of the sampling
personnel is of great importance.
7.1.5 The effect of moisture content for air dried soil samples seems to be negligible in
this case.
7.1.6 The uncertainty of the analytical process can contribute the dominating
proportion to the combined measurement uncertainty (e.g. for cadmium). It can be
controlled if the standard methods of analytical quality assurance are adhered to (e.g.
periodical use of CRMs and participation on inter-laboratory comparisons).
Uncertainty from this source may be dominant when the analyte concentration is close
to the analytical detection limit.
7.1.7 Effects that were not considered in this case study include the duration and
extent of the forces within the grinding and sieving process, and the wetness of the
soil body during the sampling process. The influence of these effects was considered
not to be significant, although these assumptions should be verified.

8 Assessment of fitness for purpose of these measurements

For a routine measurement according to the sampling protocol one composite sample
from approximately 10 increments must be analysed in duplicate.
In this case study for estimation of uncertainty contributions from single effects, 10
additional increments are taken and 20 (composite) samples are prepared and analysed
in total.
This additional effort and cost is not appropriate for routine measurements. However,
if measurements on arable land are the main type of investigation conducted by the
laboratory, such an exploratory investigation might be valuable for getting a typical
value of the sampling error component for these measurements. Furthermore, an
evaluation of the error components (i.e. uncertainty budget) will also be useful to
optimise the measurement process.

9 Reporting and interpretation

Measurements of the mean concentration for this area of top soil have expanded
uncertainty values that can be expressed as either 0.06 mg kg-1 or 18.2% of the
concentration value for cadmium, and 26 mg kg-1or 22.6% for phosphorus.

10 Summary

Measurement uncertainty*
Analyte Sampling Analytical Total
Cd 15.0% 10.4% 18.2%
P 11.6% 19.4% 22.6%
* with coverage factor of 2 (i.e. for 95% confidence)

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 Appendix B: Terminology

Appendix B: Terminology

Accuracy The closeness of agreement between a test result and the accepted
reference value.

Note: The term accuracy, when applied to a set of test results, involves a
combination of random components and a common systematic error or
bias component.

ISO 3534-1: 3.11 (1993) [9]

Bias The difference between the expectation of the test result and an
accepted reference value.

Note: Bias is a measure of the total systematic error as contrasted to

random error. There may be one or more systematic error components
contributing to the bias. A larger systematic difference from the accepted
reference value is reflected by a larger bias value.

ISO 3534-1: 3.13 (1993) [9]

Composite sample Two or more increments/sub-samples mixed together in appropriate
(also average and portions, either discretely or continuously (blended composite
aggregate) sample), from which the average value of a desired characteristic
may be obtained.

ISO 11074-2: 3.10 (1998) [45], AMC (2005) [50]

Duplicate One of the two (or more*) samples or sub-samples obtained
(replicate) sample separately at the same time by the same sampling procedure or sub-
sampling procedure. *for replicate sample

Note: Each duplicate sample is obtained from a separate sampling point

within the sampling location.

Adapted from ISO 11074-2: 2.14 (1998) [45], ISO 1998 was
formally adapted from ISO 3534-1 (1993) [9], AMC (2005) [50]
Error of result The test result minus the accepted reference value (of the
characteristic).

Note: Error is the sum of random errors and systematic errors.

ISO 3534-1: 3.8 (1993) [9]

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 Appendix B: Terminology
Fitness for purpose The degree to which data produced by a measurement process
enables a user to make technically and administratively correct
decisions for a stated purpose.

Note: As defined for analytical science.

Thompson and Ramsey (1995) [24]

Homogeneity, The degree to which a property or constituent is uniformly
heterogeneity distributed throughout a quantity of material.

Notes:
1. A material may be homogeneous with respect to one analyte or
property but heterogeneous with respect to another.
2. The degree of heterogeneity (the opposite of homogeneity) is the
determining factor of sampling error.

IUPAC (1990) [46]; ISO 11074-2: 1.6 (1998) [45]

Increment Individual portion of material collected by a single operation of a
sampling device.

IUPAC (1990) [46], AMC (2005) [50]

Laboratory sample Sample as prepared for sending to the laboratory and intended for
inspection or testing.

ISO Standard 78-2 (1999) [47]

Measurand Particular quantity subject to measurement.

ISO-GUM (1993) [2]

Measurement see Uncertainty of measurement
Uncertainty

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 Appendix B: Terminology
Precision The closeness of agreement between independent test results
obtained under stipulated conditions.

Notes:
1. Precision depends only on the distribution of random errors and does
not relate to the true value or the specified value.
2. The measure of precision usually is expressed in terms of imprecision
and computed as a standard deviation of the test results. Less precision is
reflected by a lower standard deviation.
3. Independent test results means results obtained in a manner not
influenced by any previous result on the same or similar test object.
Quantitative measures of precision depend critically on the stipulated
conditions. Repeatability and reproducibility conditions are particular
sets of extreme stipulated conditions.

ISO 3534-1: 3.14 (1993) [9]

Primary sample The collection of one or more increments or units initially taken
from a population.

Note: The term primary, in this case, does not refer to the quality of the
sample, rather the fact that the sample was taken during the earliest stage
of measurement.

IUPAC (1990) [46], AMC (2005) [50]

Random error of A component of the error which, in the course of a number of test
result results for the same characteristic, remains constant or varies in an
unpredictable way.

Note: It is not possible to correct for random error.

ISO 3534-1: 3.9 (1993) [9]

Random sample A sample of n sampling units taken from a population in such a way
that each of the possible combinations of n sampling units has a
particular probability of being taken.

ISO 3534-1: 4.8 (1993) [9]

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 Appendix B: Terminology
Random sampling; The taking of n items from a lot of N items in such a way that all
simple random possible combinations of n items have the same probability of being
sampling chosen.

Notes:
1. Random selection can never be replaced by ordinary haphazard or
seemingly purposeless choice; such procedures are generally insufficient
to guarantee randomness.
2. The phrase random sampling applies also to sampling from bulk or
continuous materials but its meaning requires specific definition for each
application.

ISO 7002: A.34 (1986) [48]

Reference sampling Characterisation of an area, using a single sampling device and a
single laboratory, to a detail allowing the set-up of a distribution
model in order to predict element concentrations, with known
uncertainty, at any sampling point.

IUPAC (2005) [49]

Reference sampling The analogue in sampling of a reference material or certified
target reference material (in chemical analysis).

Note: A sampling target, one or more of whose element concentrations

are well characterised in terms of spatial/time variability. The analogue
in sampling of a reference material or a certified reference material (in
chemical analysis) (notes adapted from IUPAC (2003) draft
recommendations; originally defined in ISO Guide 30: 1992).

Thompson and Ramsey (1995) [24]

Representative Sample resulting from a sampling plan that can be expected to
sample reflect adequately the properties of interest in the parent population.

IUPAC (1990) [46], ISO 11074-2: 1.9 (1998) [45], AMC (2005)
[50]
Sample A portion of material selected from a larger quantity of material.

IUPAC (1990) [46], ISO 11074-2 (1998) [45], AMC (2005) [50]

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 Appendix B: Terminology
Sample preparation The set of material operations (such as reduction of sizes, mixing,
dividing etc.) that may be necessary to transform an aggregated or
bulk sample into a laboratory or test sample.

Note: The sample preparation should not, as far as possible, modify the
ability of the sample to represent the population from which it was taken.

Adapted from ISO 3534-1: 4.30 (1993) [9]

Sample Collective noun for all procedures used for conditioning a sample to
pre-treatment a defined state which allows subsequent examination or analysis or
long-term storage.

Adapted from ISO 11074-2: 6.1 (1998) [45]

Sample size Number of items or the quantity of material constituting a sample.

ISO 11074-2: 4.26 (1998) [45], ISO 7002: A.40 (1986) [48]
Sampler Person (or group of persons) carrying out the sampling procedures
at the sampling point.

Note: The term sampler does not refer to the instrument used for
sampling, i.e. the sampling device.

Adapted from ISO 11074-2 (1998) [45]

Sampling Process of drawing or constituting a sample.

Note: For the purpose of soil investigation sampling also relates to the
selection of locations for the purpose of in situ testing carried out in the
field without removal of material (from ISO 1998).

ISO 11074-2 (1998) [45], ISO 3534-1 (1993) [9]

Sampling bias The part of the total measurement bias attributable to the sampling.

AMC (2005) [50]

Sampling location The place where sampling occurs within the sampling target.
Perhaps used for location within which duplicate (or replicate)
samples are taken at particular sampling points.
Sampling plan Predetermined procedure for the selection, withdrawal,
preservation, transportation and preparation of the portions to be
removed from a population as a sample.
IUPAC (1990) [46], ISO 11074-2 (1998) [45], AMC (2005) [50]

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 Appendix B: Terminology
Sampling point The place where sampling occurs within the sampling location.
Perhaps used for specific point where duplicate (or replicate)
sample taken, within a sampling location.

Note: The accuracy at which a sampling point is located in space or time

depends on the surveying method. Duplicate samples are taken from
sampling points that reflect this accuracy.
Sampling precision The part of the total measurement precision attributable to the
sampling.

AMC (2005) [50]

Sampling Operational requirements and/or instructions relating to the use of a
procedure particular sampling plan; i.e. the planned method of selection,
withdrawal and preparation of sample(s) from a lot to yield
knowledge of the characteristic(s) of the lot.

ISO 3534-1: 4.5 (1993) [], ISO 11704-2 [45] (in part), adopted by
AMC (2005) [50]
Sampling target Portion of material, at a particular time, that the sample is intended
to represent.

Notes:
1. The sampling target should be defined prior to designing the sampling
plan.
2. The sampling target may be defined by Regulations (e.g. lot size).
3. If the properties and characteristics (e.g. chemical composition) of the
certain area or period are of interest and must be known then it can be
considered a sampling target.

AMC (2005) [50]

Sampling see Uncertainty from sampling
uncertainty
Sub-sample A sample taken from a sample of a population.

Notes:
1. It may be selected by the same method as was used in selecting the
original sample, but need not be so.
2. In sampling from bulk materials, sub-samples are often prepared by
sample division. The sub-sample thus obtained is also called a divided
sample.

ISO 3534-1: 4.8 (1993) [9]

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 Appendix B: Terminology
Sub-sampling Process of selection of one or more sub-samples from a sample of a
(sample division) population.

ISO 11074-2 (1998) [45]

Systematic error of A component of the error which, in the course of a number of test
result results for the same characteristic, remains constant or varies in a
predictable way.

Note: Systematic errors and their causes may be known or

unknown.

ISO 3534-1: 3.10 (1993) [9]

Systematic Sampling by some systematic method.
sampling

ISO 3534-1: 4.15 (1993) [9], ISO 11074-2 [45]

Test portion Quantity of material, of proper size for measurement of the
concentration or other property of interest, removed from the test
sample.

IUPAC (1990) [46], ISO 11074-2: 3.17 (1998) [45], AMC (2005)
[50]
Test sample Sample, prepared from the laboratory sample, from which the test
portions are removed for testing or analysis.

IUPAC (1990) [46], ISO 11074-2: 3.16 (1998) [45], AMC (2005)
[50]
Trueness The closeness of agreement between the average value obtained
from a large series of test results and an accepted reference value.

Notes:
1. The measure of trueness is usually expressed in terms of bias.
2. The trueness has been referred to as accuracy of the mean. This usage
is not recommended.

ISO 3534-1: 3.12 (1993) [9]

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 Appendix B: Terminology
Uncertainty Parameter, associated with the result of a measurement, that
(of measurement) characterises the dispersion of the values that could reasonably be
attributed to the measurand.

Notes:
1. The parameter may be, for example, a standard deviation (or a given
multiple of it), or the half width of an interval having a stated level of
confidence.
2. Uncertainty of measurement comprises, in general, many components.
Some of these components may be evaluated from the statistical
distribution of the results of series of measurements and can be
characterised by experimental standard deviations. The other
components, which can also be characterised by standard deviations, are
evaluated from assumed probability distributions based on experience or
other information.
3. It is understood that the result of the measurement is the best estimate
of the value of the measurand, and that all components of uncertainty,
including those arising from systematic effects, such as components
associated with corrections and reference standards, contribute
dispersion.
4. (added) If measurand is defined in terms of the quantity within the
sampling target, then uncertainty from sampling is included within
uncertainty of measurement.

ISO GUM: B.2.18 (1993) [2]

Uncertainty from The part of the total measurement uncertainty attributable to
sampling sampling.
Note. Also called sampling uncertainty
IUPAC (2005) [49]

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 Appendix C: Useful statistical procedures

Appendix C: Useful statistical procedures

C1. Estimating bias between two sampling methods, by using paired samples
The paired-sample method is effected by collecting one sample, according to both of
the sampling protocols under consideration, from each of a number ( n > 20
preferably) of targets. The method is especially suitable for comparing a new
candidate protocol against an established protocol in routine use, but is also generally
applicable. For each method the sampling procedure has to be randomised in some
fashion, for example by starting the collection of increments at a random position
within the target and orientating the increment grid in a random direction. The
samples collected are analysed under randomised repeatability conditions, so that
analytical bias is cancelled out.
The design, shown below in Figure C1.1, ensures a minimum of extra work at each
target, so that the experiment can be executed at low cost without interrupting the
flow of routine sampling. The result is also rugged, because it is derived from data
collected from many typical but different targets. It therefore represents the average
bias between the results of the two protocols, rather than the bias encountered in a
single target, which may turn out to be atypical.

Figure C1.1: Design of experiment to estimate the bias between two sampling
methods

Design of experiment to estimate the bias between two sampling methods A and B, by collecting paired
samples at each target.

The first stage of the examination of the results is to check whether the paired
differences are dependent on the concentration of the analyte. This is particularly
likely to happen if the concentration range encountered in successive targets is wide.
A scatterplot provides a useful visual check. Where there is no dependence, the bias
estimate is the mean of the signed paired differences and this mean can be tested for
significant difference from zero in the usual fashion. In the example shown in Figure
C1.2, there is no apparently significant dependence between the signed difference and
the concentration, and the bias between the methods is not significantly different from
zero at the 95% level of confidence by the two-sample t-test. Where there is a clear
bias that is dependent on concentration, as in Figure C1.3, the bias should be
expressed as a function of concentration. In the instance illustrated, there is evidence
(established by the functional relationship method [40]) of a significant rotational bias
with a trend expressed by the equation Result (B) = Result (A) 1.2.
.
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 Appendix C: Useful statistical procedures
Figure C1.2: No significant bias or trend

Differences between results of two sampling protocols applied to 25 targets, as a function of the
concentration. There is no significant bias and no suggestion of a dependence of bias on concentration.

Figure C1.3: Significant bias and trend

Differences between results of two sampling protocols applied to 33 targets, plotted as a function of the
concentration. There is a significant bias (because 27/33 results are negative) and the absolute bias
increases with increasing concentration.

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 Appendix C: Useful statistical procedures

C2. Further description of sampling errors from sampling theory

C2.1 Weighting error (SWE) forms its own class. It is created, for example, if the lot
(sampling target) consists of sub-lots of different sizes but the mean concentration is
estimated as a simple mean, without taking the sizes of the sub-lots into account. The
correct method is to calculate the weighted mean by using the sizes of the sub-lots as
weights. In analysis of moving material, weighting error is generated if the flow-rate
varies but is not taken into account in calculating the mean; in this case the flow-rates
should be recorded simultaneously with sampling and used as weights in calculating
the mean. Another option is to use a sampling device that cuts samples whose size is
proportional to the flow-rate and use the sample sizes as weights in calculating the
mean. It should be noted that if a composite sample is made from sub-samples then
proportional sampling should be used; otherwise a weighting error is generated in the
composite sample.
C2.2 Grouping and segregation error (GSE) is the second error term related to
short range errors. It is caused by the fact that the sample is normally not taken
fragment by fragment, but as a group of fragments. If there is segregation in the
material, this causes this type of error. This error is not normally estimated. Gy has
shown, however, that if the sampling is correctly done GSE is smaller than, or at
maximum equal to, the fundamental sampling error (FSE).
C2.3 Point selection error (PSE). When the mean of a continuous object (e.g.
process stream, river, polluted site, ...) is estimated by using discrete samples, the
uncertainty of the mean depends on the sampling strategy, because the results are
usually autocorrelated. This error is called point selection error (PSE) and it depends
on the sampling strategy. Three basic strategies can be applied for picking the samples
(see Figure C2.1):
1) Random sampling: Time or location of the sampling points are randomly
distributed along the target.
2) Stratified (random) sampling: The lot is first divided into N sub-lots of equal
sizes and within each sub-lot the sampling point is randomly assigned.
3) Systematic (stratified) sampling: All N samples are collected at equal
distances (one-dimensional case) or on a fixed symmetric pattern (targets
which from the sampling point of view have two or more dimensions).

Estimation of the standard deviation of the mean of the lot

sp
Random sampling: s (a L ) =
N
s strat
Stratified sampling: s (a L ) =
N
s sys
Systematic sampling: s (a L ) =
N
sstrat and ssys are standard deviation estimates, where the autocorrelation has been
taken into account.

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 Appendix C: Useful statistical procedures
Normally the order is sp > sstrat > ssys, except when in systematic sampling the
sampling frequency is a multiple of process frequency. In this case the systematic
sampling is the worst choice and the mean may be biased.

Figure C2.1: Sampling strategies

100
CONCENTRATION

50
Random selection
0
0 5 10 15 20 25 30
TIME
CONCENTRATION

100

50
Stratified selection
0
0 5 10 15 20 25 30
TIME
CONCENTRATION

100

50
Systematic selection
0
0 5 10 15 20 25 30
TIME

Ten samples selected from the target by using random, stratified random and
systematic stratified sample selection.

Estimation of the PSE

The distribution heterogeneity of a one-dimensional lot can be characterised by
carrying out a variographic experiment, i.e. N samples are collected from the target by
using systematic sample selection. N should be at least 30, preferably 60100.
Proportional cross-stream sampling should be used or if not possible (when large gas
or liquid streams are sampled) the flow-rate should be recorded simultaneously with
the sampling time. From these results the experimental heterogeneity hi can be
calculated as the relative variation about the lot mean (or mean of the sampling
target). When N samples of size Mi are collected and analysed (results are ai). Mi can
be also the flow-rate, if proportional sampling cannot be carried out.
ai a L M i
hi = ( i =1, 2,K, N )
aL M
where aL is the weighted mean of the lot:

M a 1 Mi
aL =
i i
= ( M )a
M
i
i N
The standard deviation of the heterogeneity h is equal to the relative standard
deviation of the lot or process, sp.

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 Appendix C: Useful statistical procedures
To characterise the variability of the process an experimental variogram is calculated
from the heterogeneities:

( )
N j
1 N
Vj = hi hi
2( N j ) i = 1 + j
2
, j = 1,2, K ,
2
The variogram has to be integrated to estimate the PSE for different sampling
strategies. Gy uses a robust numerical integration.

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 Appendix C: Useful statistical procedures

C3. Sources of software for calculations

Classical analysis of variance (ANOVA) is available in most general spreadsheet

software for one-way ANOVA. F-tests and other standard statistical tests for the
normal distribution are also implemented in most spreadsheets.

Programs for general robust statistical methods in general, and for robust ANOVA in
particular, are available from RSC/AMC
(http://www.rsc.org/Membership/Networking/InterestGroups/Analytical/AMC/Softwa
re/index.asp).

Outlier tests (e.g. Grubbs or Dixons) are less generally available, as is software for
the range method. The range method can, however, be implemented relatively simply
using maximum and minimum functions in a spreadsheet.
The range calculations (demonstrated in Section 7 of Appendix A3) are easily
performed using standard spreadsheets, and an example can be downloaded from
http://team.sp.se/analyskvalitet/sampling/default.aspx.

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 Appendix D: Alternative experimental designs for empirical uncertainty
estimation

Appendix D: Alternative experimental designs for empirical

uncertainty estimation

1. Multi-level designs to estimate other component effects

The general balanced design for the empirical estimation of uncertainty (Figure 2)
includes the uncertainty from the physical sample preparation in with the sample
step. An alternative experimental design (Figure D.1) can be used to make a separate
estimate of the uncertainty from this source (sprep). Two sub-samples from both of the
two primary samples are prepared separately (grey boxes in Figure D.1). Duplicate
test portions are taken from these sub-samples so that the analytical contribution can
also be estimated. The standard robust ANOVA can be used to separate all of these
sources of variance (Figure A1.2, and Appendix C3), by selecting two different sub-
sets of four measurements, shown in Figure D.1. Full details of the application of this
design to food sampling are given elsewhere [30].

Figure D.1: Experimental design utilised for the estimation of uncertainty from
sample preparation, as well as that from sampling and analysis

Sampling

Sample 1 Sample 2
(S1) (S2)
2kg 2kg

Prep. 1 Prep. 2 Prep. 1 Prep. 2

(P1) (P2) (P1) (P2)
1kg 1kg 1kg 1kg

Analysis 1 Analysis 2 Analysis 1 Analysis 2 Analysis 1 Analysis 1

(A1) (A2) (A1) (A2) (A1) (A1)

S1P1A1 S1P1A2 S1P2A1 S1P2A2

S1P1A1 S1P2A1 S2P1A1 S2P2A1

* The upper section depicts the three-layered and unbalanced experimental design. The additional layer in this experimental
design, required for the evaluation of sprep, is shown by the grey boxes. The lower section (shaded) shows the data groupings
required for the application of ANOVA so as to provide estimates of ssamp, sprep and sanal, i.e. the statistical design. Figure taken
from [30] with permission of Royal Society of Chemistry.

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 Appendix D: Alternative experimental designs for empirical uncertainty
estimation

2. Simplified and unbalanced designs, to reduce the cost of implementation

Figure D.2 Two simplified alternatives to the full balance design (Figure 2) that
can be applied to reduce the cost of estimating the measurement uncertainty
using the empirical approach: (a) the simplified balanced design, and (b) the
unbalanced design

Sampling Target

(a)
Sample 1 Sample 2

Measurement Uncertainty

Analysis 1 Analysis 1

Sampling Target
(b)

Sampling Uncertainty Sample 1 Sample 2

Analytical Uncertainty Analysis 1 Analysis 2 Analysis 1

The simplified design (Figure D.2a) has the same duplicated samples as in the full
balanced design (Figure 2), but does not include and duplicated chemical analyses.
The uncertainty estimated using this design gives the total measurement uncertainty,
without any values for the components of the uncertainty from the sampling or the
analysis. If these components are required, the analytical uncertainty can be estimated
externally by the laboratory, and removed from the total uncertainty, to give a
separate estimate of the sampling uncertainty, using Equation 1. The main advantage
of this design is that the analytical cost of implementation is only half of that for the
full balanced design, for the same number of duplicated samples. Alternatively, twice
the number of duplicated samples can be taken, from twice the number of targets to
increase their representativeness for the same expenditure on chemical analysis.
The unbalanced design (Figure D.2b) is intermediate between these two designs, with
only one analytical duplicate carried out on one of the duplicated samples. This has
the advantage of giving estimates of the sampling and analytical components of the
uncertainty, as well as the total measurement uncertainty (with the same caveats
expressed as for the full balanced design in Section 9.4.2). The analytical costs are
reduced by 25% compared with those for the fully balanced case. The degrees of
freedom in this case are similar for both the analytical and sampling estimates of
variance, which is more cost-effective than the extra degrees of freedom for the
analytical uncertainty in the fully balanced case.
Classical ANOVA can be applied to the output of both of these designs using many
different spreadsheet software packages (Appendix C3), but robust ANOVA has not
yet been developed for this case.

UfS:2007 Page 98
 Appendix E: Modifying sampling uncertaitny using predictions from sampling
theory

Appendix E: Modifying sampling uncertainty using predictions from

sampling theory
Once uncertainty from sampling has been estimated, and if it is found not to be fit for
purpose, there may be a need to modify this level of uncertainty. Predictions on how
to achieve this modification can be made using sampling theory (Section 10.2).
Several theories predict that the sampling variance is inversely proportional to the
mass of the sample taken (e.g. Equation 5). This leads to the prediction that any
required modification of uncertainty of sampling (from usamp1 to usamp2) can be
calculated by changing the mass of the sample (from ms1 to ms2) using the relationship
ms2 = (usamp1 / usamp2)2 . ms1 ..(Equation E1)
This approach can usefully be illustrated using the case study of nitrate in lettuce in
Example A1. The sampling uncertainty was shown not to be fit for purpose (by the
method in Section 16.3), and the optimal uncertainty required was calculated to be
lower by a factor of approximately 2. Equation E1 predicts that this should be
achieved by increasing the sample mass by a factor of 4 (i.e. 22). The implementation
of this prediction by increasing the number of increments from 10 heads to 40 heads
of lettuce per batch, did achieve the predicted reduction in the sampling uncertainty in
this case (i.e. by a factor of 1.80, which is not statistically significantly different from
the predicted improvement of 2.0) [38]. Such successful predictions are not always
achieved in practice. In a different example for the determination of moisture in
butter, a predicted reduction of 3.7 in the usamp, was calculated to require an increase
in ms by a factor of 14. In practice this increase in sample mass only produced an
experimental improvement of 1.3. The inability of this model to predict the change in
sampling uncertainty was probably due to the nature of the heterogeneity of the
analytes in this particular material [51].

UfS:2007 Page 99
 Appendix F: References

Appendix F: References
1 Ellison S L R, Roesslein M, Williams A (eds) (2000) Eurachem/CITAC Guide: Quantifying
Uncertainty in Analytical Measurement, Eurachem, 2nd edition, ISBN 0 948926 15 5. Available from
the Eurachem secretariat, or from LGC Limited (London).
2 ISO (1993) Guide to the Expression of Uncertainty in Measurement (GUM). Geneva (2nd printing
1995).
3 Ellison S L R, Roesslein M, Williams A (eds) (1995) Eurachem Guide: Quantifying Uncertainty in
Analytical Measurement, Eurachem, 1st edition.
4 Analytical Methods Committee (1995) Uncertainty of measurement: implications of its use in
analytical science. Analyst, 120, 23032308.
5 ISO/TS 21748 (2004) Guidance for the use of repeatability, reproducibility and trueness estimates in
measurement uncertainty estimation. ISO, Geneva.
6 Gy P M (1979) Sampling of Particulate Materials Theory and Practice. Elsevier, Amsterdam,
431pp.
7 Codex (2004) General Guidelines on Sampling. CAC/GL-2004
(http://www.codexalimentarius.net/download/standards/10141/CXG_050e.pdf)
8 Nordtest (2007) Uncertainty from sampling. A Nordtest handbook for sampling planners and
sampling quality assurance and uncertainty estimation. NT tec 604/TR604 (www.nordicinnovation.net)
9 ISO 3534-1: 1993 Statistics Vocabulary and Symbols, International Organization for
Standardization, Geneva
10 Ramsey M H, Squire S, Gardner M J (1999) Synthetic reference sampling target for the estimation
of measurement uncertainty. Analyst, 124 (11), 17011706.
11 Ramsey M H (1998) Sampling as a source of measurement uncertainty: techniques for
quantification and comparison with analytical sources. Journal of Analytical Atomic Spectrometry, 13,
97104.
12 Lyn J A, Ramsey M H, Coad D S, Damannt A P, Wood R, Boon K A The duplicate method of
uncertainty estimation: are 8 targets enough? (Submitted to Analyst)
13 De Zorzi P, Belli M, Barbizzi S, Menegon S, Deluisa A (2002) A practical approach to assessment
of sampling uncertainty. Accreditation and Quality Assurance, 7, 182188.
14 Kurfurst U, Desaules A, Rehnert A, Muntau H (2004) Estimation of measurement uncertainty by the
budget approach for heavy metal content in soils under different land use. Accreditation and Quality
Assurance, 9, 6475.
15 Minkkinen P (2004) Practical applications of sampling theory. Chemometrics and Intelligent Lab.
Systems, 74, 8594.
16 Gy P M (1998) Sampling for Analytical Purposes. John Wiley & Sons Ltd, Chichester.
17 Gy P M (1992) Sampling of Heterogeneous and Dynamic Material Systems. Elsevier, Amsterdam.
18 Gy P M (2004) Proceedings of First World Conference on Sampling and Blending. Special Issue of
Chemometrics and Intelligent Lab. Systems, 74, 770.
19 Pitard F F (1993) Pierre Gy's Sampling Theory and Sampling Practice. CRC Press, Boca Raton,
2nd edition.
20 Smith P L (2001) A Primer for Sampling Solids, Liquids and Gases Based on the Seven Sampling
Errors of Pierre Gy. ASA SIAM, USA.
21 Ramsey M H (1993) Sampling and analytical quality control (SAX) for improved error estimation
in the measurement of heavy metals in the environment, using robust analysis of variance. Applied
Geochemistry, 2, 149153.
22 Nordtest Sampler Certification, Version 1-0, Nordic Innovation Centre (2005).
23 Thompson M, Wood R (1995) Harmonised guidelines for internal quality control in analytical
chemistry laboratories. Pure and Applied Chemistry, 67, 649666.

UfS:2007 Page 100

 Appendix F: References
References (Continued)
24 Thompson M, Ramsey M H (1995) Quality concepts and practices applied to sampling an
exploratory study. Analyst, 120, 261270.
25 Magnusson B, Naykki T, Hovind H, Krysell M (2003) Handbook for Calculation of Measurement
Uncertainty. NORDTEST report TR 537 (project 1589-02). Can be obtained from
www.nordicinnovation.net/nordtest.cfm under link Rapporter.
26 Ramsey M H, Squire S, Gardner M J (1999) Synthetic reference sampling target for the estimation
of measurement uncertainty. Analyst, 124 (11), 17011706.
27 Thompson M, Coles B J, Douglas J K (2002) Quality control of sampling: proof of concept.
Analyst, 127, 174177.
28 Farrington D, Jervis A, Shelley S, Damant A, Wood R, Thompson M (2004) A pilot study of routine
quality control of sampling by the SAD method, applied to packaged and bulk foods. Analyst, 129,
359363.
29 Codex (2004) Guidelines on measurement uncertainty CAC/GL 54-2004.
30 Lyn J A, Ramsey M H, Fussel RJ, Wood R (2003) Measurement uncertainty from physical sample
preparation: estimation including systematic error. Analyst, 128 (11), 1391 1398
31 Uncertainty and Assessment of Compliance Eurachem (in preparation).
32 Measurement Uncertainty and Conformance Testing: Risk Analysis. Joint Committee on General
Metrology, Working group 1/Subcommittee 3 (in preparation).
33 Ramsey M H, Thompson M, Hale M (1992) Objective evaluation of precision requirements for
geochemical analysis using robust analysis of variance. Journal of Geochemical Exploration, 44, 23
36.
34 Thompson M, Fearn T (1996) What exactly is fitness for purpose in analytical measurement?
Analyst, 121, 275278.
35 Commission Regulation (EC) No 563/2002 of 2 April 2002 amending Regulation (EC) No
466/2001 setting maximum levels for certain contaminants in foodstuffs, Official Journal of the
European Communities, L 86/5 to L 86/6.
36 European Directive 79/700/EEC. OJ L 207, 15.8.1979, p26.
37 Ramsey M H, Lyn J A, Wood R (2001) Optimised uncertainty at minimum overall cost to achieve
fitness-for-purpose in food analysis. Analyst, 126, 17771783.
38 Lyn, J A, Palestra I M, Ramsey M H, Damant A P, Wood R (2007) Modifying uncertainty from
sampling to achieve fitness for purpose: a case study on nitrate in lettuce. Accreditation and Quality
Assurance: Journal for Quality, Comparability and Reliability in Chemical Measurement, 12, 67-74.
39 Thompson M, Walsh J N A (1989) Handbook of Inductively Coupled Plasma Spectrometry.
Blackie, Glasgow.
40 Analytical Methods Committee (2002) Technical Brief No.10, Royal Society of Chemistry, London.
41 Ramsey M H, Taylor P D, Lee J C (2002) Optimized contaminated land investigation at minimum
overall cost to achieve fitness-for-purpose. Journal of Environmental Monitoring, 4 (5), 809814.
42 Lee J C, Ramsey M H (2001) Modelling measurement uncertainty as a function of concentration: an
example from a contaminated land investigation. Analyst, 126 (10), 17841791.
43 Ramsey M H, Argyraki A (1997) Estimation of measurement uncertainty from field sampling:
implications for the classification of contaminated land. Science of the Total Environment, 198, 243
257.
44 Dahinden R, Desaules A (1994) Die Vergleichbarkeit von Schwermetallanalysen in Bodenproben
von Dauerbeobachtungsflchen, Eidgenssische Forschungsanstalt fr Agrikulturchemie und
Umwelthygiene. Liebefeld, Switzerland, p27.
45 ISO 11074-2: 1998 Soil Quality Vocabulary. Part 2: Terms and definitions related to sampling,
International Organization for Standardization, Geneva.
46 IUPAC (1990) Nomenclature for sampling in analytical chemistry (Recommendations 1990),
prepared for publication by Horwitz W, Pure and Applied Chemistry, 62, 11931208.

UfS:2007 Page 101

 Appendix F: References
References (Continued)

47 ISO Standard 78-2: Chemistry Layouts for Standards Part 2: Methods of Chemical Analysis
(Second Edition, 1999).
48 ISO 7002: 1986 Agricultural food products Layout for a standard method of sampling from a lot,
First Edition, International Organization for Standardization, Geneva.
49 IUPAC (2005) Terminology in Soil Sampling (IUPAC Recommendations 2005), prepared for
publication by De Zorzi P, Barbizzi S, Belli M, Ciceri G, Fajgelj A, Moore D, Sansone U, and Van der
Perk M. Pure and Applied Chemistry, 77 (5), 827841.
50 AMC (2005) Analytical Methods Committee Technical Brief No 19. Terminology the key to
understanding analytical science. Part 2: Sampling and sample preparation
(http://www.rsc.org/images/brief19_tcm18-25963.pdf).
51 Lyn J A, Ramsey M H, Damant A, Wood R (2005), Two stage application of the OU method: a
practical assessment, Analyst, 130, 1271-1279

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