General Topics:

UNIT 15 GENERAL TOPICS: RELATED TO Related to Food

Testing Laboratories
FOOD TESTING LABORATORIES
Structure
15.0 Objectives
15.1 Introduction
15.2 Method Validation
15.2.1 When a Method should be Validated?
15.2.2 Who is Responsible for Ensuring Validation of a Test Method?
15.2.3 What should be Extent of Validation?
15.2.4 The Method Performance Parameters for a Test Method
15.2.5 Confirmation of Identity and Selectivity/Specificity
15.3 Ruggedness (or Robustness)
15.3.1 Determination
15.3.2 Recovery
15.4 Uncertainty of Measurement
15.4.1 What is Uncertainty?
15.4.2 How does it Arise?
15.4.3 Significance of Uncertainty in Analytical Measurements
15.4.4 What is Done about it?
15.4.5 Calibration Field
15.4.6 Testing Field
15.5 International Accreditation Aspects
15.5.1 Evaluation of Uncertainty
15.5.2 What is Best Practice?

15.6 Statements of Compliance - Effect of Uncertainty

15.6.1 Definition of Uncertainty
15.6.2 Quantity under Measurement
15.6.3 Uncertainty Sources
15.6.4 Uncertainty Components
15.6.5 Error and Uncertainty
15.7 Reference Materials
15.7.1 Types of Reference Materials
15.7.2 Desirable Reference Materials/Certified Reference Materials
15.8 Proficiency Testing
15.8.1 Inter Laboratory Test Comparisons
15.8.2 Proficiency Testing
15.8.3 Official Definitions
15.9 Why Proficiency Testing are Essential?
15.10 Different Types of Proficiency Test Schemes/Program
15.10.1 Essential Components of Proficiency Testing Programs
15.10.2 Availability of Proficiency Test Schemes/Programs
15.10.3 International Scene
15.10.4 National Scene

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Laboratory Quality 15.11 Codex Food Control Laboratory
Management System
15.12 Let Us Sum Up
15.13 Key Words
15.14 Answers to Check your Progress Exercise
15.15 Suggested Reading

15.0 OBJECTIVES
After reading this unit, we shall be able to:
• describe the importance of Method validation in the field of analysis and
apply the same for residue testing of food products;
• understand about uncertainty of measurement in testing and attempt to
apply for some simple analytical techniques; and
• understand the importance and necessity of participating in Proficiency
testing for a food testing laboratory learn about the requirements of Codex
Food Control laboratory.

15.1 INTRODUCTION
There are certain general concepts which are associated with different types of
analysis falling within the gamut of food testing. These are:
1) Method validation,
2) Measurement of Uncertainty,
3) Certified reference materials, and
4) Proficiency testing.
This Unit will make us familiar with these concepts which are very important
from the point of view of Quality Management systems in any food testing
laboratory and personnel from laboratories intending to implement systems as
per ISO 17025 must be familiar with.

15.2 METHOD VALIDATION

For analytical results to be fit for its intended purpose they must be sufficiently
reliable so that any decision based on it can be taken with confidence. It is not
only important to determine the correct result but also to be able to
demonstrate that it is correct and is fit for the purpose’. Method validation
enables chemists to demonstrate that a test method is ‘fit for purpose’.
Method validation is the process of defining an analytical requirement and
conforming that the method under consideration has performance capabilities
consistent with what the application requires. In the validation process, the
objective is to estimate the representativeness, repeatability and reproducibility
of the test method and to secure adequate evidence that the method is fit for its
intended purpose.
The Standardized test methods published by national and international
standards bodies (IS, ISO/IEC, DIN, etc.) or independent technical bodies
(ASTM, AOAC, IAEA, etc.), or regulatory bodies (IP, BP, EPA) should be
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considered validated for their intended applications (range and accuracy of General Topics:
measurement) and thus considered fit for intended purpose even if repeatability Related to Food
Testing Laboratories
and reproducibility are not known in detail. However the laboratory should
ensure that they apply the method correctly.

15.2.1 When a Method should be Validated?

A method needs to be validated under the following circumstances :
a) New method developed for particular problem:
Lab developed method or
Non-standard method
b) Standard methods used outside of their intended scope.
c) Amplification and modification of standard methods.
d) Established method revised to incorporate improvements or extended to a
new problem.
e) When quality control indicates an established method is changing with
time.
f) To demonstrate the equivalence between two methods, a new method and a
standard method.
Validation may appropriately include procedures for sampling, handling and
transportation.

15.2.2 Who is Responsible for Ensuring Validation of a Test

Method?
a) Standards Body publishing the test method.
b) Lab to establish performance data for their own use of the method, where
necessary.
c) Laboratory using a method is responsible for ensuring that the method used
is validated.
d) Method performance claims must be validated by a laboratory.

15.2.3 What should be Extent of Validation?

The extent of validation or revalidation required will depend on the nature of
the changes made in reapplying a standard method. Starting with a carefully
considered analytical specification provides a good base for planning the
validation process. A balance need to be struck between the need and extent of
validation and the efforts required to be put in terms of time and cost. The
design of method validation plan /protocol should normally cover the method
performance parameters needed to be characterised for validation.

15.2.4 The Method Performance Parameters for a Test Method

These are:
• selectivity/specificity and Confirmation of identity,
• Limit of detection,
• Limit of quantification,
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Laboratory Quality • Working and linear ranges,
Management System
• Recovery (Accuracy / trueness),
• Precision (Repeatability and Reproducibility), and
• Robustness and Ruggedness.

15.2.5 Confirmation of Identity and Selectivity/Specificity

It is the ability of a test method to determine accurately and specifically the
analyte of interest in the presence of other components in a sample matrix
under the stated conditions of the test.
a) Selectivity in qualitative terms is the extent to which other analytes
interfere with the determination of an analyte in accordance with a given
test procedure and in quantitative terms it is a constant, coefficient, index,
factor or a number used for the quantitative characterisation of
interferences.
For an analytical method it is essential to establish that the signal produced
at the measurement stage or any other measured property, which has been
attributed to the analyte, is only due to the analyte and not from the
presence of some other similar entities or present by chance. The extent of
interference from the interfering entity generally depends on the
effectiveness of the isolation stage and the selectivity/specificity of the
measurement stage. For example in pesticide residue analysis of food
products using chromatography technique the extent of interference would
depend on the extraction process and the selectivity and specificity on the
type of detector used. Mass detectors would be the most specific/selective.
Selectivity and specificity are measures which assess the reliability of
measurements in the presence of interferences. Where the measurement
stage is non-specific, and the interferences are known then it is possible to
state in the test method that certain analytes interfere, so that the analyst
can take appropriate precautions. At times it is desirable that in such case
measurement may be followed by confirmation of analyte identity by
independent techniques.
The selectivity of a method is usually investigated by studying its ability to
measure the analyte of interest in test portions to which specific
interferences have been deliberately introduced (those thought likely to be
present in samples).
Where it is unclear whether or not interferences are already present, the
selectivity of the method can be investigated by studying its ability to
measure the analyte compared to other independent methods / techniques.
b) Limit of Detection (LOD)
In trace analysis it is important to know what is the lowest concentration of
the analyte or property value that can be confidently detected by the
method. LOD of a test method is the lowest concentration of analyte in a
sample that can be detected, but not necessarily quantified under the
stated conditions of the test.
Mean and the standard deviation of the sample blank are likely to be
dependant on the matrix of the sample blank. Limit of detection will
therefore be matrix dependent.

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 For qualitative measurements LOD is defined as the concentration General Topics:
threshold below which specificity becomes unreliable. The threshold may Related to Food
Testing Laboratories
vary if the experiment is repeated at another time with different reagents,
fortification, spiking materials, etc.
c) Limit of Quantitation (LOQ)
The Limit of quantitation (LOQ) is the lowest concentration of analyte that
can be determined with an acceptable level of repeatability precision and
trueness under the stated conditions of the test.
Sometimes, LOQ is also known as ‘limit of determination’, ‘limit of
Reporting’.
Limit of Quantification (LOQ) – The Content equal to or greater than the
lowest concentration point on the calibration curve - AOAC Definition
d) Working & Linear Ranges
For any quantitative method, it is necessary to determine the range of
analyte concentrations over which the method can be applied. At the lower
end of the concentration range the limiting factors are the LOD and/or
LOQ values.
At the upper end of the concentration range limitations will be imposed by
various effects depending on the instrument response system, they could
also be in terms of saturation of detector. In a complex method like residue
analysis it will be a combination of analyte quantity in matrix, the
extraction efficiency and instrument (detection) sensitivity.
Within the working range there may exist a linear response range. Within
the linear range signal response will have a linear relationship to analyte
concentration. The extent of this range may be established during the
evaluation of the working range.
Analysts should keep following aspects in mind while working on an
analytical method:
a) In general, linearity checks would require 10 points corresponding to
10 different concentrations.
b) Evaluation of the working and linear ranges may be required on a day-
to-day basis if they are directly linked to calibration.
c) Within the linear range, 1-3 calibration point may be sufficient, to
establish the slope of the calibration line. Elsewhere in the working
range, multi-point (preferably 6+) calibration would be necessary.
d) The relationship of instrument response to concentration does not have
to be perfectly linear for a method to be effective but the curve should
be repeatable from day to day.
e) The working and linear range may be different for different matrices
according to the effect of interferences arising from the matrix.
e) Accuracy
Accuracy expresses the closeness of a result to a true value. Method
validation seeks to quantify the likely accuracy of results by assessing both
systematic and random effects on results. Accuracy is, therefore, normally
studied as two components: ‘trueness’ and ‘precision’ The Trueness (of a
method) is an expression of how close the mean of a set of results

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Laboratory Quality (produced by the method) is to the true values. Trueness is generally
Management System
assessed comparison of mean results from a method with known values.
Trueness is assessed against a reference value (true value or conventional
true value). Two basic techniques for estimation are checking against
reference values of CRM or from another standard method.
Precision is a measure of how close results are to one another and is
usually expressed by measures such as standard deviation, which describe
the spread of results. Precision is normally determined for specific
circumstances which in practice can be very varied.
The two most common precision measures are repeatability and
reproducibility.
An increasingly common expression of accuracy is ‘measurement
uncertainty’, which provides a single figure expression of accuracy.
f) Sensitivity
This is effectively the gradient of the response curve, i.e. the change in
instrument response which corresponds to a change in analyte
concentration. Where the response has been established as linear with
respect to concentration, i.e. within the linear range of the method, and the
intercept of the response curve has been determined, sensitivity is a useful
parameter to calculate and use in formulae for quantitation. Sensitivity is
sometimes used to refer to limit of detection but this use is not generally
approved, because in a method the limit of detection is dependent on many
parameters and instrument sensitivity alone.

15.3 RUGGEDNESS (OR ROBUSTNESS)

A measure of an effectiveness of an analytical method is how well its
performance stands up to less than perfect implementation. In any method,
there will be certain stages which, if not carried out sufficiently carefully, will
have a severe effect on method performance and may even result in the method
not working at all.
These stages should be identified, usually as part of method development, and
if possible, their influence on method performance evaluated using
‘ruggedness tests’, sometimes also called ‘robustness tests’.

15.3.1 Determination
This involves making deliberate variations to the method, and investigating the
subsequent effect on performance.

It is then possible to identify the variables in the method which have the most
significant effect and ensure that, when using the method, they are closely
controlled. Where there is a need to improve the method further, improvements
can probably be made by concentrating on those parts of the method known to
be critical. Ruggedness is normally evaluated during method development,
typically by the originating laboratory, before collaborating with other
laboratories.
Ruggedness tests are normally applied to investigate the effect on either
precision or accuracy.

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15.3.2 Recovery General Topics:
Related to Food
Testing Laboratories
Analytical methods generally do not require the analyst to always measure all
of the analyte of interest present in the sample. Analyst may be interested in
only one type of analyte and may choose a method which is designed to
determine only the analyte of interest. It is necessary to assess the efficiency of
the method in detecting the analyte interest as present in deferent matrices.
This determination of efficiency of the method (recovery) can easily be done
using a sample of known analyte concentration, which is the best way of doing
so. However such reference samples are not easily available. Although this
method is not as good as use of reference samples, the difficulty can be
overcome by preparing a synthetic mixture by spiking test portions with the
analyte at various concentrations, then extracting the fortified test portions and
measure the analyte concentration.
The techniques used for determination of performance of a method could be
one or combinations of the following:
a) Calibration using reference standards or reference material;
b) Comparison of results achieved with other methods;
c) Systematic assessment of factors influencing result;
d) Assessment of the uncertainty of results based on scientific understanding
of the theoretical principles and practical experience; and
e) Inter laboratory comparison.

15.4 UNCERTAINTY OF MEASUREMENT

Although the results of analytical measurements are very significant when
decisions are required to be made based on test / analysis results, it is an
accepted fact that true value of a measurand (a quantity subject to
measurement) is indeterminate, except when known in terms of theory. What is
normally obtained as a result of measurement process is at best an estimate of
or approximation to the true value. Even when appropriate corrections for
known or suspected components of error have been applied, there still remains
an uncertainty, that is, a doubt about how well the result of measurement
represents the true value of the quantity being measured. This fact would not
normally inspire much confidence.

15.4.1 What is Uncertainty?

However, when used in a technical sense as in 'measurement uncertainty' or
'uncertainty of a test result' it carries a specific meaning. It is a parameter,
associated with the result of a measurement (e.g. a calibration or test) that
defines the range of the values that could reasonably be attributed to the
measured quantity. When uncertainty is evaluated and reported in a specified
way it indicates the level of confidence that the value actually lies within the
range defined by the uncertainty interval. Normally the uncertainty of
measurement is a parameter that characterizes the dispersion of the true
values which could reasonably be attributed to the measurand. The parameter
may be, for example, the standard deviation or a given multiple of it, or the
half-width of an interval having a stated level of confidence. Hence when one
is certain about the uncertainty, one can make appropriate conclusion/
decision.
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Laboratory Quality 15.4.2 How does it Arise?
Management System
Any measurement is subject to imperfections; some of these are due to
random effects, such as short-term fluctuations in temperature, humidity and
air-pressure or variability in the performance of the measurer. Repeated
measurements will show variation because of these random effects. Other
imperfections are due to the practical limits to which correction can be made
for systematic effects, such as offset of a measuring instrument, drift in its
characteristics between calibrations, personal bias in reading an analogue
scale or the uncertainty of the value of a reference standard.

15.4.3 Significance of Uncertainty in Analytical Measurements

The uncertainty is a quantitative indication of the quality of the result. It gives
an answer to the question - how well does the result represent the value of the
quantity being measured? It allows users of the result to assess its reliability,
for example for the purposes of comparison of results from different sources or
with reference values. In fact, it is expected that a statement of results of
measurement (as a process) is complete only if it contains both the values.
Without such an indication, measured results cannot be compared, either
among themselves or with reference values given in a specification or
standard. Confidence in the comparability of results can help to reduce barriers
to trade.
Often, a result is compared with a limiting value defined in a specification or
regulation. In this case, knowledge of the uncertainty shows whether the result
is well within the acceptable limits or only just makes it. Occasionally, a result
is so close to the limit that the risk associated with the possibility that the
property that was measured may not fall within the limit, once the uncertainty
has been allowed for, must be considered.
Suppose that a customer has the same test done in more than one laboratory,
perhaps on the same sample, more likely on what they may regard as an
identical sample of the same product. Would we expect the laboratories to get
identical results? Only within limits, in actual practice! But when the results
are close to the specification limit, it may be that one laboratory indicates
failure, whereas another indicates a pass. Now what the user/customer of the
test result to do? In cases like this an appropriate decision would have been
facilitated, if the uncertainty of the result had been known by the customer.

15.4.4 What is Done about it?

The standard ISO/IEC 17025: 2005 (General requirements for the competence
of testing and calibration laboratories) specifies requirements for reporting
and evaluating uncertainty of measurement. The problems presented by these
requirements vary in nature and severity depending on the technical field and
whether the measurement is a calibration or test.

15.4.5 Calibration Field

Calibration is characterised by the facts that:
1) Repeated measurements can be made,
2) Uncertainty of reference instruments is provided at each stage down the
calibration chain, starting with the national standard, and

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3) Customers are aware of the need for a statement of uncertainty in order to General Topics:
ensure that the instrument meets their requirements. Related to Food
Testing Laboratories
Consequently, calibration laboratories are used to evaluating and reporting
uncertainty. In accredited laboratories, the uncertainty evaluation is subject to
assessment by the accreditation body and is quoted on calibration certificates
issued by the laboratory.

15.4.6 Testing Field

The situation in testing is not as well-developed and many difficulties,
particular to different testing areas are encountered. For example, in
destructive tests the opportunity to repeat the test is limited to another sample,
often at significant extra cost and with the additional uncertainty due to
sample variation. Even when repeat tests are technically feasible such an
approach may be uneconomic. In some cases a test may not be defined well
enough by the standard, leading to potentially inconsistent application and
thus another source of uncertainty. In many tests, there will be uncertainty
components that need to be evaluated on the basis of previous data and
experience, in addition to those evaluated from calibration certificates and
manufacturers, specifications.
a) Requirement as per ISO 17025:2005
i) Estimation of uncertainty of measurement.
ii) A calibration laboratory, or a testing laboratory performing its own
calibrations, shall have and shall apply a procedure to estimate the
uncertainty of measurement for all calibrations and types of
calibrations.
iii) Testing laboratories shall have and apply procedures for estimating
uncertainty of measurement. In certain cases, the nature of the test
method may preclude rigorous, metrologically and statistically valid,
calculation of uncertainty of measurement. In these cases, the
laboratory shall at least attempt to identify all the components of
uncertainty and make a reasonable estimation, and shall ensure that the
form of reporting of the results does not give a wrong impression of
the uncertainty. Reasonable estimation shall be based on knowledge of
the performance of the method and on the measurement scope and
shall make use of, for example, previous experience and validation
data.
NOTE 1: The degree of rigor needed in an estimation of uncertainty of
measurement depends on factors such as:
• The requirements of the test method.
• The requirements of the client.
• The existence of narrow limits on which decisions on conformation
to a specification are based.
NOTE 2: In those cases, where a well recognised test method specifies
limits to the values of major sources of uncertainty of measurement
and specifies the form of presentation of calculated results, the
laboratory is considered to have satisfied the clauses following the test
method and reporting instructions.

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Laboratory Quality iv) When estimating the uncertainty of measurement, all uncertainty
Management System
components which are of importance in the given situation shall be
taken into account using appropriate methods of analysis.
NOTE 1: Sources contributing to the uncertainty include, but are not
necessarily limited to the reference standards and reference materials
used, methods and equipment used, environmental conditions,
properties and conditions of the item being tested or calibrated, and the
chemist.
NOTE 2: The predicted long term behaviour of the tested and/or
calibrated item is not normally taken into account when estimating the
measurement uncertainty.

15.5 INTERNATIONAL ACCREDITATION

ASPECTS
Accreditation bodies are responsible for ensuring that accredited laboratories
meet the requirements of ISO/IEC 17025. The standard requires appropriate
methods of analysis to be used for estimating uncertainty of measurement.
These methods are considered to be those based on the Guide to the
expression of uncertainty of measurement, published by ISO and endorsed by
the major international professional bodies. It is a weighty document and the
international accreditation community has taken up its principles and along
with other bodies such as EURACHEM/CITAC, has produced simplified or
more specific guidance based on them. EURACHEM/CITAC Guide CG 4 is
most relevant for analytical Measurements, and forms the basis for this
training program.

15.5.1 Evaluation of Uncertainty

Uncertainty is a consequence of the unknown sign of random effects and
limits to corrections for systematic effects and is therefore expressed as a
quantity, i.e. an interval about the result. It is evaluated by combining a
number of uncertainty components. The components are quantified either by
evaluation of the results of several repeated measurements or by estimation
based on data from records, previous measurements, knowledge of the
equipment and experience of the measurement.
In most cases, repeated measurement results are distributed about the average
in the familiar bell-shaped curve or normal distribution, in which there is a
greater probability that the value lies closer to the mean than to the extremes.
The evaluation from repeated measurements is done by applying a relatively
simple mathematical formula. This is derived from statistical theory and the
parameter that is determined is the standard deviation.
Uncertainty components quantified by means other than repeated
measurements are also expressed as standard deviations, although they may
not always be characterised by the normal distribution. For example, it may be
possible only to estimate that the value of a quantity lies within bounds (upper
and lower limits) such that there is an equal probability of it lying anywhere
within those bounds. This is known as a rectangular distribution. There are
simple mathematical expressions to evaluate the standard deviation for this
and a number of other distributions encountered in measurement. An
interesting one that is sometimes encountered, e.g. in EMC measurements, is
the U-shaped distribution.
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The method of combining the uncertainty components is aimed at producing a General Topics:
realistic rather than pessimistic combined uncertainty. This usually means Related to Food
Testing Laboratories
working out the square root of the sum of the squares of the separate
components (the root sum square method). The combined standard uncertainty
may be reported as it stands (the one standard deviation level), or usually, an
expanded uncertainty is reported. This is the combined standard uncertainty
multiplied by what is known as a coverage factor. The greater this factor the
larger the uncertainty interval and correspondingly, the higher the level of
confidence that the value lies within that interval. For a level of confidence of
approximately 95%, a coverage factor of 2 is used. When reporting
uncertainty it is important to indicate the coverage factor or state the level of
confidence, or both.

15.5.2 What is Best Practice?

Use of sector-specific procedures for estimation of uncertainty is considered a
good practice. However such sector specific guidance, especially in testing
fields is still not available in several fields to enable laboratories to evaluate
uncertainty consistently. Further even though the practice of reporting
uncertainty with the test results is not prevalent in testing field, laboratories
are encouraged to evaluate uncertainty in order to assess the quality of their
own results, especially when the results are close to any specified limit. The
process of evaluation highlights those aspects of a test or calibration that
produce the greatest uncertainty components, thus indicating where
improvements could be beneficial. Conversely, it can be seen whether larger
uncertainty contributions could be accepted from some sources without
significantly increasing the overall interval. This could give the opportunity to
use cheaper, less sensitive equipment or provide justification for extending
calibration intervals.
Uncertainty evaluation is best done by personnel who are thoroughly familiar
with the test or calibration and understand the limitations of the measuring
equipment and the influences of external factors, e.g. environment. Records
should be kept showing the assumptions that were made, e.g. concerning the
distribution functions referred to above, and the sources of information for the
estimation of component uncertainty values, e.g. calibration certificates,
previous data, experience of the behaviour of relevant materials.

15.6 STATEMENTS OF COMPLIANCE - EFFECT

OF UNCERTAINTY
This is a difficult area and what is to be reported should be considered in the
context of the client's needs. In particular, consideration should be given to the
possible consequences and risks associated with a result that is close to the
specification limit. The uncertainty may be such as to raise real doubt about
the reliability of pass/fail statements. When uncertainty is not taken into
account, then the larger the uncertainty, the greater are the chances of passing
failures and failing passes. A lower uncertainty is usually attained by using
better equipment, better control of environment, and ensuring consistent
performance of the test.
For some products it may be appropriate for the user to make a judgment of
compliance, based on whether the result is within the specified limits with no
allowance made for uncertainty. This is often referred to as shared risk, since
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Laboratory Quality the end user takes some of the risk of the product not meeting specification.
Management System
The implications of that risk may vary considerably. Shared risk may be
acceptable in non-safety critical performance, for example the EMC
characteristics of a domestic radio or TV. However, when testing a heart
pacemaker or components for aerospace purposes, the user may require that
the risk of the product not complying has to be negligible and would need
uncertainty to be taken into account. An important aspect of shared risk is that
the parties concerned agree on the uncertainty that is acceptable; otherwise
disputes could arise later.

In Conclusion Uncertainty is an unavoidable part of any measurement and it

starts to matter when results are close to a specified limit. A proper evaluation
of uncertainty is good professional practice and can provide laboratories and
customers with valuable information about the quality and reliability of the
result. Although common practice in calibration, there is some way to go with
expression of uncertainty in testing, but there is growing activity in the area
and in time, uncertainty statements will be the norm.

15.6.1 Definition of Uncertainty

The definition of the term uncertainty (of measurement) as taken from the
current version adopted for the International Vocabulary of Basic and General
Terms in Metrology is:

“A parameter associated with the result of a measurement, that characterises

the dispersion of the values that could reasonably be attributed to the
measurand”.
Note 1: The parameter may be, for example, a standard deviation (or a
given multiple of it), or the width of a confidence interval.
Note 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 standard deviations. The other components,
which also can be characterised by standard deviations, are evaluated
from assumed probability distributions based on experience or other
information. The ISO Guide refers to these different cases as Type A
and Type B estimations respectively.

15.6.2 Quantity under Measurement

In many cases in chemical analysis, the measurand will be the concentration*
of an analyte. However chemical analysis is also carried out to measure other
quantities, e.g. colour, pH, etc., and therefore the general term "measurand" is
normally used for describing the quantity under measurement.

The definition of uncertainty given above focuses on the range of values that
the analyst believes could reasonably be attributed to the measurand.
* The unqualified term “concentration” applies to any of the particular quantities mass
concentration, amount concentration, number concentration or volume concentration unless
units are quoted (e.g. a concentration quoted in mg l-1 is evidently a mass concentration).
Further many other quantities used to express composition, such as mass fraction, substance
content and mole fraction, can be directly related to concentration.

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In general use, the word uncertainty relates to the general concept of doubt. In General Topics:
this guide, the word uncertainty, without adjectives, refers either to a parameter Related to Food
Testing Laboratories
associated with the definition above, or to the limited knowledge about a
particular value. Uncertainty of measurement does not imply doubt about the
validity of a measurement on the contrary, knowledge of the uncertainty
implies increased confidence in the validity of a measurement result.

15.6.3 Uncertainty Sources

In practice the uncertainty on the result may arise from many possible sources,
including examples such as incomplete definition, sampling, matrix effects and
interferences, environmental conditions, uncertainties of masses and
volumetric equipment, reference values, approximations and assumptions
incorporated in the measurement method and procedure, and random variation
(a fuller description of uncertainty sources is given in subsequent chapters).

15.6.4 Uncertainty Components

In estimating the overall uncertainty, it may be necessary to take each source
of uncertainty and treat it separately to obtain the contribution from that
source. Each of the separate contributions to uncertainty is referred to as an
uncertainty component. When expressed as a standard deviation, an
uncertainty component is known as a standard uncertainty. If there is
correlation between any components then this has to be taken into account by
determining the covariance. However, it is often possible to evaluate the
combined effect of several components. This may reduce the overall effort
involved and where components whose contribution is evaluated together are
correlated, there may be no additional need to take account of the correlation.
For a measurement result y, the total uncertainty, termed as combined
standard uncertainty and denoted by uc(y), is an estimated standard deviation
equal to the positive square root of the total variance obtained by combining all
the uncertainty components, however evaluated, using the law of propagation
of uncertainty.
For most purposes in analytical chemistry, an expanded uncertainty U,
should be used. The expanded uncertainty provides an interval within which
the value of the measurand is believed to lie with a higher level of confidence.
U is obtained by multiplying uc(y), the combined standard uncertainty, by a
coverage factor k. The choice of the factor k is based on the level of
confidence desired. For an approximate level of confidence of 95%, k is 2.
NOTE: The coverage factor k should always be stated so that the combined
standard uncertainty of the measured quantity can be recovered for
use in calculating the combined standard uncertainty of other
measurement results that may depend on that quantity.

15.6.5 Error and Uncertainty

It is important to distinguish between error and uncertainty. Error is defined as
the difference between an individual result and the True value of the measured.
As such, error is a single value. In principle, the value of a known error can be
applied as a correction to the result.
NOTE: Error is an idealised concept and errors cannot be known exactly,
especially since true value itself is an indeterminate value.
101
Laboratory Quality Uncertainty, on the other hand, takes the form of a range, and, if estimated for
Management System
an analytical procedure and defined sample type, may apply to all
determinations so described. In general, the value of the uncertainty cannot be
used to correct a measurement result.
To illustrate further the difference, the result of an analysis after correction
may by chance be very close to the value of the measured, and hence have a
negligible error. However, the uncertainty may still be very large, simply
because the analyst is very unsure of how close that result is to the value.
The uncertainty of the result of a measurement should never be interpreted as
representing the error itself, nor the error remaining after correction.
An error is regarded as having two components, namely, a random component
and a systematic component.
Random error typically arises from unpredictable variations of influence
quantities. These random effects give rise to variations in repeated
observations of the measured. The random error of an analytical result cannot
be compensated for, but it can usually be reduced by increasing the number of
observations.
NOTE: The experimental standard deviation of the arithmetic mean or
average of a series of observations is not the random error of the
mean, although it is so referred to in some publications on
uncertainty. It is instead a measure of the uncertainty of the mean due
to some random effects. The exact value of the random error in the
mean arising from these effects cannot be known.
Systematic error is defined as a component of error which, in the course of a
number of analysis of the same measured, remains constant or varies in a
predictable way. It is independent of the number of measurements made and
cannot therefore be reduced by increasing the number of analyses under
constant measurement conditions.
Constant systematic errors, such as failing to make an allowance for a reagent
blank in an analysis, or inaccuracies in a multi-point instrument calibration, are
constant for a given level of the measurement value but may vary with the
level of the measurement value.
Effects which change systematically in magnitude during a series of analyses,
caused, for example by inadequate control of experimental conditions, give
rise to systematic errors that are not constant.
EXAMPLES:
1) A gradual increase in the temperature of a set of samples during a chemical
analysis can lead to progressive changes in the result.
2) Sensors and probes that exhibit ageing effects over the time-scale of an
experiment can also introduce non-constant systematic errors.
The result of a measurement should be corrected for all recognised significant
systematic effects.
NOTE: Measuring instruments and systems are often adjusted or calibrated
using measurement standards and reference materials to correct for
systematic effects. The uncertainties associated with these standards
and materials and the uncertainty in the correction must still be taken
into account.
102
A further type of error is a spurious error, or blunder. Errors of this type General Topics:
invalidate a measurement and typically arise through human failure or Related to Food
Testing Laboratories
instrument malfunction. Transposing digits in a number while recording data,
an air bubble lodged in a spectrophotometer flow through cell, or accidental
cross-contamination of test items are common examples of this type of error.
Measurements for which errors such as these have been detected should be
rejected and no attempt should be made to incorporate the errors into any
statistical analysis. However, errors such as digit transposition can be corrected
(exactly), particularly if they occur in the leading digits.
Spurious errors are not always obvious and, where a sufficient number of
replicate measurements is available, it is usually appropriate to apply an outlier
test to check for the presence of suspect members in the data set. Any positive
result obtained from such a test should be considered with care and, where
possible, referred back to the originator for confirmation. It is generally not
wise to reject a value on purely statistical grounds.
The procedures used for estimation of Uncertainties generally do not to allow
for the possibility of spurious errors/blunders.

15.7 REFERENCE MATERIALS

The reliability of analytical/measurement data play a very important role in the
decision making process involved in commerce and trade and in verifying
conformity to various regulations, health and safety aspects or reciprocal
comparisons. Reference materials (RM’s) are very important for realising a
number of aspects of measurement quality and reliability which help in correct
decision making.
Reference Materials are required for use in any one or more of the following
activities which are essential components of a measurement process:
a) Calibration of Instrument;
b) Standardisation of analytical measurement;
c) Method validation studies;
d) Estimation of uncertainty of measurement;
e) Training of analysts / testing personnel;
f) Internal quality control; and
g) External quality assurance (QA) (proficiency testing).
Availability of authentic and credible reference Materials (Reference Material
with certified value - CRM) is one of the very important factors affecting
laboratory’s capability to produce reliable test results.
In an international comparison/competitiveness scenario brought about by
accreditation, it is of utmost importance that test results produced by different
accredited labs are comparable and use of reliable reference materials plays
one of the most important roles in achieving this.
In fact in most of modern methods of chemical analysis using techniques like
Chromatography, spectroscopy/spectrometry, instrumentation involving mass
detectors, etc., the results are directly related to reference materials which are
used for calibration/standardization of instruments. Hence the need for
reference material is the highest in chemical analysis involving such types of
techniques.
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Laboratory Quality As per ISO definition (as given in ISO-Guides 30:1992 ) Reference Material
Management System
(RM) is a Material or substance one or more of whose property values are
sufficiently homogeneous and well established to be used for the calibration of
an apparatus, the assessment of a measurement method, or for assigning values
to materials.
Another term which is used in relation to Reference materials is Certified
Reference Materials (CRM) which is defined in ISO-Guides 30:1992 as:
Reference material, accompanied by a certificate, one or more of whose
property values are certified by a procedure which establishes its traceability to
an accurate realization of the unit in which the property values are expressed,
and for which each certified value is accompanied by an uncertainty at a stated
level of confidence.
Most results of analytical measurements are directly related to reference
materials which are used for the calibration of the measurement process. For
fulfilling the requirements of the laboratory accreditation criteria ISO
17025:2005, two decisive preconditions are required to be met in order to
obtain a result which is traceable to the SI. First, the used reference material
itself must carry an SI-traceable value and an attached uncertainty. Secondly,
the whole measurement procedure - from sampling to calculation of the result -
must be fully validated and the uncertainty must be evaluated according to
common rules. While the analyst is required to validate his procedure and to
evaluate his measurement uncertainty by himself, he has to depend on
reference material supplier to ensure that the value declared on the label of the
reference material is traceable to the SI. Thus, the reference material plays a
very important role as it serves as a transfer standard in the traceability chain.
And in a laboratory accreditation scenario certified reference material
(Reference Material with a valid certificate) is an essential component.

15.7.1 Types of Reference Materials

RMs may be used to support measurements concerned with chemical
composition, biological, clinical, physical, engineering properties, etc. They
may be characterised for ‘identity’ (e.g. chemical structure, fibre type,
microbiological species/cultures etc.) or for ‘property values’ (e.g. amount of
specified chemical entity, hardness etc.). Some commonly encountered
categories of reference materials are as follows:
a) Chemical Composition - Reference materials, being either pure chemical
compounds or representative sample matrices, either natural (e.g. metals)
or with added analytes (e.g. animal fats spiked with pesticides for residues
analysis), characterised for one or more chemical or physicochemical
property values;
b) Biological and Clinical Properties - Materials similar to above category,
but characterised for one or more biochemical or clinical property values;
c) Physical Properties - Materials characterised for one or more physical
property values, e.g. melting point, viscosity, density;
d) Engineering Properties - Materials characterised for one or more
engineering property values (e.g. hardness, tensile strength, surface
characteristics, electrical resistance, etc.;
e) Miscellaneous Materials.

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15.7.2 Desirable Reference Materials/Certified Reference General Topics:
Related to Food
Materials Testing Laboratories

a) It must be uniform and homogenous in composition and pure in cases

where relevant;
b) The exact property value must be known;
c) Traceability to SI units should be established;
d) Its shelf life (if relevant) must be known;
e) The Measurement Uncertainty of the property value must be known; and
f) It should be available in fairly large quantities for effective distribution.
In addition to the above for Certified Reference Material, it is desirable that:
a) The reference value for relevant property/characteristics is known and is
certified by a procedure which establishes its traceability to an accurate
realization of the unit in which the property values are expressed.
b) The validity of the ‘certification’ and uncertainty data, including
conformance of key procedures should be established in line with the
requirements of ISO Guide 35 and other relevant ISO requirements.
c) Both the producer and the material should have a track record of good
performance, for example when - a RM in use has been subjected to an
inter-laboratory comparison, cross-checked by use of different methods, or
there is experience of use in a number of laboratories over a period of
years.
d) Each CRM should be accompanied by a certificate stating the property
value and an uncertainty value at a stated level of confidence in line with
the requirements of ISO Guide 31 and a report covering the
characterisation, certification and statistical analysis procedures, complying
with ISO Guide 35.
e) There should be a demonstrated compliance of the production of the
reference materials with quality standards such as ISO Guides 34 or ILAC
requirements, or compliance of the measurement of property values with
ISO/IEC 17025 requirements.

15.8 PROFICIENCY TESTING

A variety of means are adopted by the laboratories for monitoring the
validity/reliability of test data generated and the performance of tests
undertaken, which include – Replicate tests using same or different test
methods, Retesting/recalibration of retained items, correlation of results,
comparison of test results with other laboratories through participation in Inter
laboratory test comparison or Proficiency testing programs, etc. These are also
prescribed in ISO 17025:2005 as a means of ensuring quality of testing by
laboratories. Among those listed above, participation in Proficiency testing
programs are generally independent assessments, hence most preferred.
Even the laboratory accreditation systems as per ISO/IEC 17025, recommend a
regular independent assessment of the technical performance of a laboratory,
as an important means of assuring the validity of analytical measurements and
also as a part of the overall quality strategy.
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Laboratory Quality 15.8.1 Inter Laboratory Test Comparisons
Management System
Inter laboratory Test Comparisons are conducted for a number of purposes and
may be used by participating laboratories and other parties for the following
purposes:
a) To determine the performance of individual laboratories for specific tests
or measurements and to monitor laboratories' continuing performance.
b) To identify problems in laboratories and initiate remedial actions, which
may be related, for example, to individual staff performance or calibration
of instrumentation, etc.
c) To establish the effectiveness and comparability of new test or
measurement methods and similarly to monitor established methods.
d) To provide additional confidence to laboratory customers.
e) To identify Inter laboratory differences.
f) To determine the performance characteristics of a test method - often
known as collaborative trials, Inter laboratory trials, round-robins to
determine its fitness for use.
g) For assigning values to reference materials (RMs) and assess their
suitability for use in specific test or measurement procedures as part of
CRM production.

15.8.2 Proficiency Testing

It is the use of Inter laboratory comparisons to determine the performance of
individual laboratories for specific tests or measurements.
How is it Done?
Inter-comparisons of test results can be organized by the laboratory itself and
involve only a few other laboratories (Inter laboratory test comparison), or the
laboratory can participate in an independent inter-comparison services
provided by a designated lab or professional provider (PT provider), where
usually a number of other laboratories participate.
A Proficiency Testing (PT) scheme/program is a system for objectively
evaluating laboratory results by external means, and includes regular
comparison of a laboratory’s results at intervals with those of other
laboratories. This is achieved by the scheme/program coordinator regularly or
through one-time distribution of homogeneous test samples to participating
laboratories for analysis and reporting of the data. Each distribution of test
samples is referred to as a round. The main objective of a PT scheme is to help
the participating laboratory to assess the accuracy of its test results.

15.8.3 Official Definitions

Inter laboratory comparison (ILC) is the organisations performance and
evaluation of calibrations/tests on the same or similar calibration/test items by
two or more laboratories in accordance with predetermined conditions.
Proficiency testing (PT) is the determination of the calibration or testing
performance of a laboratory or the testing performance of an inspection body
by means of inter laboratory comparison.

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 General Topics:
15.9 WHY PROFICIENCY TESTING ARE Related to Food
ESSENTIAL? Testing Laboratories

While individual laboratories use Inter laboratory comparisons/PT Program

data for internal verifications, accreditation bodies like the NABL use
proficiency testing as part of the overall assessment of a laboratory’s ability to
perform tests competently. It is increasingly considered as an essential part of
the accreditation program, complementing the on-site laboratory assessment by
technical experts, and providing information to support the accreditation
decision process. In many countries, in certain specialized areas, which are
generally in the regulated sector, like food testing, drinking water and
environmental testing, participation in certain specified PT programs by the
laboratories is considered essential as per the requirements stipulated in the
regulation.
In view of the above all laboratory accreditation bodies in their accreditation
criteria/system specify the minimum amount of proficiency testing (Self
initiated Inter laboratory comparison studies in absence of availability of PT
programs) and the frequency of participation, appropriate in relation to other
surveillance activities.
Further notwithstanding what is stated in ISO 17025 and the accreditation
procedure of the laboratory accreditation body, proficiency testing is an
excellent way to validate one’s own measurement/analysis processes.
Proficiency tests can validate the participating laboratory’s
measurement/analytical method, technical training of personnel, traceability of
standards and uncertainty budgets. Even though the laboratory may had been
most rigorous in developing the above mentioned laboratory processes, it is
important to use proficiency testing as an additional way to verify that all
aspects of the laboratory measurement system are sound and hence are
considered very essential.

15.10 DIFFERENT TYPES OF PROFICIENCY

TEST SCHEMES/ PROGRAM
Proficiency testing techniques vary depending on the nature of the test item,
the method in use and the number of laboratories participating. Most possess
the common feature of comparison of results obtained by one laboratory with
those obtained by one or more other laboratories. In some schemes, one of the
participating laboratories may have a controlling, coordinating, or reference
function.
The most common types of proficiency testing schemes/programs are -
Measurement comparison schemes, Inter laboratory testing schemes, Split-
sample testing scheme, Qualitative schemes, Known-value schemes, and
Partial-process schemes.

15.10.1 Essential Components of Proficiency Testing

Programs
There are two distinct processes involved in designing and organizing a
Proficiency testing program, which are concerning the following:
a) Sample or the artifact (as in case of testing of discrete articles or
calibration), and;
107
Laboratory Quality b) The statistical processing and evaluation of data for drawing accurate
Management System
conclusions on performance of participating laboratories.
Both the steps need to be designed and carried out appropriately in order to
ensure useful and accurate conclusions arising out of the Proficiency testing
program.
The following factors related to sample or the artifact have direct influence on
quality and reliability of Proficiency testing performed.
a) Sample/artifact quality, homogeneity / reliability – samples/artifacts are
needed to be carefully selected keeping in mind all essential components.
The selection process should consider the uncertainty attainable,
transportation and packaging conditions required.
b) Homogeneity of the sample/artifact – This needs to be ensured through
appropriate homogeneity studies.
c) Availability of all relevant information in respect of stability of the
sample/artifact at least from the point of characterization (including during
transportation) to the point of testing/calibration; attainable uncertainty,
etc.
d) Choice of appropriate reference laboratory, where required, for accurate
characterization (Reference Value and uncertainty determination, relevant
specially for calibration) of artifact/sample.
Other important factors which have direct influence on the conclusions drawn
on the basis of participation of proficiency testing are:
a) Statistical design of the proficiency testing program, with appropriate
choice of test statistics used. This is required to be decided at the planning
stage itself.
b) Appropriate statistical treatment of the data generated by the participating
laboratories for drawing accurate and meaningful conclusions.
c) Design and quality of final report – Important from users point of view.
d) Completion of the entire program within reasonable time frame as planned
for during the design stage and prompt delivery of report to participating
laboratories.

15.10.2 Availability of Proficiency Test Schemes/Program

With growth of laboratories and corresponding realization of importance of the
test results in decision making processes in many economic, health and safety
areas, the requirements of reliable and accurate results has grown
tremendously.
In recent years both laboratories and accreditation bodies have increasingly
realised the importance of proficiency tests as tool not only for helping the
laboratories to assess their own performance and helping them in improving
reliability of test/calibration data but also as an excellent tool to demonstrate
and assess the performance of laboratories with regard to specific
measurement, testing, and analytical tasks. Today it is also well appreciated
that Proficiency tests can support accreditation processes and indirectly, give a
feedback on the quality and impact of accreditation. Despite these benefits the
participation in proficiency tests is generally hampered not only by the
restricted offer of suitable proficiency testing schemes in many testing areas
108
but also by the lack of information on their availability. However General Topics:
internationally this scene has changed in recent years. Related to Food
Testing Laboratories

15.10.3 International Scene

Today a large number of Proficiency Test Providers are available especially
spread over in USA, Europe and Australia – some laboratories in government
sector, some association level laboratories, some are National Measurement
Institutes of different countries and many others are totally independent private
professional PT providers. Some of these have been in existence as early as in
1970’s. A very comprehensive list of PT providers world over in different test
and calibration areas is available on the website (www.nata.asn.au) of
Australian accreditation body NATA (National Association of Testing
Authorities, Australia).
In fact, in Europe, in an effort to collate information about PT providers world
over and create a network of PT providers, in 1998, 16 organisations in 16
European countries joined forces. A network was accordingly established
under the auspices of the European Co-operation for Accreditation (EA) and
the European laboratory organisations, EURACHEM and EUROLAB, in
subsequent years supported by organisation like ILAC, EA, IRMM, IAAC
and many others from USA Latin America. The partners of this network
extensively surveyed the availability of PT schemes and their characteristics
within their respective countries. All this was entered into a database which
was made public on the internet in the year 2000. A website was created to
give access to the database, and to provide additional information concerning
PT. The network, website and database were called EPTIS for European PT
Information System. The European Union supported the foundation of EPTIS
with financial contribution convinced about its contribution to strengthening
PT as a valuable proof of technical competence and its role to help removing
technical barriers to trade. After the EU funding of the project ended, the 16
partners agreed to continue their co-operation on a voluntary basis. They
formalized their cooperation in a joint Protocol in 2001.
Besides constant updating of the database a new section for PT providers who
wish to announce their forthcoming PT rounds has been installed. They also
include information regarding accredited PT programs. It is apparent from the
information contained in database that today large number of PT providers
internationally in various testing and calibration fields and their number is
growing by the day.
With a number of PT providers now available world over, in recent years the
need for performance evaluation of Proficiency test providers to assess their
fitness for purpose, by an independent body has also been felt. Many of the
laboratory accreditation bodies in USA, Europe and that in Australia have
introduced Accreditation of PT providers.

15.10.4 National Scene

The national laboratory accreditation system has been in existence in India
since 1991/92. The need for participation in PT program was primarily driven
by the requirements given in the laboratory accreditation criteria. Subsequently
the need for participation in PT programs increased with stricter norms being
prescribed as a result of revision of Accreditation criteria for laboratories
(ISO/IEC 17025:2005 and also the earlier version of 1999) and recently with
revision in Criteria for accreditation bodies (ISO/IEC 17011:2004).
109
Laboratory Quality Further to meet the requirements of APLAC MR001 it is mandatory for all
Management System
accrediting bodies, who are APLAC MRA partners, to undertake proficiency
testing programs for its accredited laboratories. All applicant laboratories are
required to successfully participate in at least one Inter Laboratory Proficiency
Testing in accordance with ISO/IEC Guide-43 (I)- 1996.
However there are hardly any independent, professional PT providers in the
country. The gap is partly met through organization of PT programs by NABL
for its accredited as well as applicant laboratories. Non-accredited laboratories
are also invited to participate in these programs. The programs are run through
nodal laboratories identified by NABL. NABL has also prepared and published
NABL - 162 Guidelines for Proficiency Testing programs for Testing and
Calibration Laboratories for conducting PT programs. Further, as advised by
NABL, accredited laboratories regularly participate in the PT programs
organized by APLAC and other international bodies.
NABL also encourages participation of applicant / accredited laboratories in
Proficiency Testing programs conducted by other national and international
bodies. Accredited laboratories are encouraged to participate in as many of
these external programs as possible to enhance their quality assurance.

15.11 CODEX FOOD CONTROL LABORATORY

Food Control Laboratory is a term generally used for describing a laboratory
which is used for testing of food products under a regulatory framework.
CODEX has prescribed guidelines for management of Food Control
Laboratories vide its standard CAC/GL 28-1995, Rev.1-1997.
Vide this standard it has recommended that following Protocols and Guidelines
should be implemented by the food control laboratories for the purpose of
Quality Assurance.
a) “International Harmonized Protocol for the Proficiency Testing of
(Chemical) Analytical Laboratories”, published by Pure & Applied
Chemistry (IUPAC), 65 (1993) 2132-2144 and J. AOAC International, 76
(1993) 926-940; The contents of the harmonised protocol include main
topics like Definitions, Organisation (Protocol) of Proficiency Testing
Schemes, Generalised Statistical Procedure for the Analysis of Results -
Approaches to Data Analysis in Proficiency.
b) “Protocol for the Design, Conduct and Interpretation of Method
Performance Studies”, published by Pure & Applied Chemistry (IUPAC),
67 (1995) 331-343 – The protocol covers definition of all related terms. It
also includes details on Design of Method performance study, statistical
analysis and system of reporting.
c) “Harmonized Guidelines for Internal Quality Control in Analytical
Chemistry Laboratories”, Published by Pure & Applied. Chem., 67 (1995)
649-666 - The topics covered are Definitions, Quality assurance practices
and Internal quality control, Internal quality control procedures, IQC and
within run precision, Control materials in IQC, etc.

# Check Your Progress Exercise 1

Note: a) Use the space below for your answers.
b) Compare your answers with those given at the end of the unit.

110
1) What are the desirable characteristics of reference materials/certified General Topics:
reference material? Related to Food
Testing Laboratories
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
2) Explain two essential components of proficiency testing program?
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….

3) What is uncertainty and how does it arise?

…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
4) List the method performance parameters for a test method?
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….
…………………………………………………………………………….

15.12 LET US SUM UP

Essential components of Laboratory accreditation system, which will ensure
reliability of analytical data, are Method Validation, Uncertainty of
Measurement and Proficiency Testing. There is a great significance and use of
method validation in chemical residue analysis in food products. The extent of
validation or revalidation required will depend on the nature of the changes
made in applying a standard method. Starting with a carefully considered
analytical specification provides a good base for planning the validation
process. A balance need to be struck between the need and extent of validation
and the efforts required to be put in terms of time and cost. The design of
method validation plan/protocol should normally cover the method
performance parameters needed to be characterised for validation. Proficiency
testing and its significance in laboratory accreditation and different types of
proficiency testing schemes have been discussed here. The most common types
of proficiency testing schemes/programs are - Measurement comparison
schemes, Inter laboratory testing schemes, Split-sample testing scheme,
Qualitative schemes, Known-value schemes, Partial-process schemes. What is
uncertainty of measurement and what are essential components of its
estimation, evaluation of effect of uncertainty on statement of compliance all
have been elaborated here. Essential features of Codex Food Control
111
Laboratory Quality Laboratory - a laboratory which is used for testing of food products under a
Management System
regulatory framework is also discussed in this unit.

15.13 KEY WORDS

Accuracy : The accuracy of a method is expressed as
the closeness of an obtained result to a
true value.
Calibration : Calibration is a set of operations that
establish, under specified conditions, the
relationship between values of quantities
indicated by a measuring instrument or
measuring system, or values represented
by a material measure, and the
corresponding values realized by
standards.
Certified Reference : Reference material, accompanied by a
Material (CRM) certificate, one or more of whose property
values are certified by a procedure, which
establishes its traceability to an accurate
realisation of the units in which the
property values are expressed, and for
which each certified value is accompanied
by an uncertainty at a stated level of
confidence.
Limit of Detection (LoD) : The lowest content that can be measured
with reasonable statistical certainty. It is
also defined as the lowest concentration of
analyte in a sample that can be detected,
but not necessarily quantified under the
stated conditions of the test.
Limit of Quantitation : It is the lowest concentration of analyte
(LoQ) that can be determined with an acceptable
level of repeatability precision and
trueness under the stated conditions of the
test. It is also defined as the Content equal
to or greater than the lowest concentration
point on the calibration curve.
Also sometimes known as ‘limit of
determination’, ‘limit of Reporting’.
Linear Range : It is the range of analyte concentrations
over which the method gives test results
proportional to the concentration of the
analyte.
Linearity : Defines the ability of the method to obtain
test results proportional to the
concentration of analyte.
Measurement : A parameter associated with the result of a
Uncertainty measurement that characterises the
dispersion of the values that could
reasonably be attributed to the measurand.
112
Method Validation : It is the confirmation by examination and General Topics:
Related to Food
through provisions of objective evidence Testing Laboratories
that the particular requirements for a
specific intended use are fulfilled (ISO
definition).
Precision : It is a measure of how close results are to
one another and is usually expressed by
measures such as standard deviation,
which describe the spread of results. The
two most common precision measures are
‘Repeatability’ and Reproducibility’.
Proficiency Testing : A recognized way for a laboratory to
(Inter Laboratory monitor its performance against both its
Comparisons) own requirements and the requirements of
Accreditation bodies
Quality Control (QC) : The operational techniques and activities
that are used to fulfill requirements for
quality. Examples of QC: Control
charting; blank determinations; spiked
samples; repeat determinations; blind
samples.
Range (Measuring- : Set of values of measurands for which the
Working) error of a measuring instrument is
intended to lie within specified limits.
Recovery : The fraction of analyte added to a test
sample (fortified or spiked sample) prior
to analysis, the unfortified and fortified
samples, percentage recovery (%R) is
calculated as follows:
%R = [(CF-CU)/CA] x 100
Where CF is the concentration of analyte
measured in the fortified sample; CU is
the concentration of analyte measured in
the unfortified sample; CA is the
concentration of analyte added (measured
value, not determined by method) in
fortified sample.
Reference Material : Material or substance one or more of
(Rm) whose property values are sufficiently
homogeneous and well established to be
used for the calibration of an apparatus,
the assessment of a measurement method,
or for assigning values to materials.
Repeatability : It is the smallest expected precision. It
gives an idea of the sort of variability to be
expected when a method is performed by a
single analyst on one piece of equipment
over a short timescale, i.e. the sort of
variability to be expected between results
when a sample is analysed in duplicate.
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Laboratory Quality
Management System
Reproducibility : It is the largest measure of precision
normally encountered. Reproducibility
gives an idea of the sort of variability to be
expected when a method is performed by
different laboratories, different analyst,
different equipment, different conditions,
over a long timescale, i.e. the sort of
variability to be expected during inter
laboratory comparisons.
Robustness: : The robustness of an analytical procedure
is a measure of its capacity to remain
unaffected by small, but deliberate
variations in method parameters and
provides an indication of its reliability
during normal usage.
Ruggedness : The ruggedness of an analytical procedure
is the measure of its capacity to remain
unaffected by variations in the
environmental and/or operating conditions
as likely to arise in different environment
during Inter laboratory studies.
Sample : A portion of material selected to represent
a larger body of material.
Sample Handling : This refers to the manipulation to which
samples are exposed during the sampling
process, from the selection from the
original material through to the disposal of
all samples and test portions.
Sample Preparation : This describes the procedures followed to
select the test portion from the sample (or
sub-sample) and includes: in-laboratory
processing; mixing; reducing; coning &
quartering; riffling; and milling &
grinding.
Selectivity/Specificity : The ability of a test method to determine
(Confirmation Of accurately and specifically the analyte of
Identity) interest in the presence of other
components in a sample matrix under the
stated conditions of the test.
Sensitivity : This is effectively the gradient of the
response curve, i.e. the change in
instrument response which corresponds to
a change in analyte concentration, where
the response has been established as linear
with respect to concentration (within the
linear range of the method) and the
intercept of the response curve has been
determined.
Sub-Sample : This refers to a portion of the sample
obtained by selection or division; an
114
 individual unit of the lot taken as part of General Topics:
the sample or; the final unit of multi-stage Related to Food
Testing Laboratories
sampling.
Test Portion : This refers to the actual material weighed
or measured for the analysis.
Traceability : Property of the result of a measurement or
the value of a standard whereby it can be
related to stated references, usually
national or international standards,
through an unbroken chain of comparisons
all having stated uncertainties.
Trueness : The ‘trueness’ (of a method) is an
expression of how close the mean of a set
of results (produced by the method) is to
the true value. Trueness is normally
expressed in terms of bias.

15.14 ANSWERS TO CHECK YOUR PROGRESS "

EXERCISE
Your answer should include following points:
Check Your Progress Exercise 1

1) a) It must be uniform and homogenous in composition and pure in cases

where relevant;
b) The exact property value must be known;
c) Traceability to SI units should be established;
d) Its shelf life (if relevant) must be known;
e) The Measurement Uncertainty of the property value must be known;
and
f) It should be available in fairly large quantities for effective distribution.

2) There are two distinct processes involved in designing and organizing a

Proficiency testing program, which are concerning the following:
a) Sample or the artifact (as in case of testing of discrete articles or
calibration), and
b) The statistical processing and evaluation of data for drawing accurate
conclusions on performance of participating laboratories. Both the steps
need to be designed and carried out appropriately in order to ensure
useful and accurate conclusions arising out of the Proficiency testing
program.

3) a) It is a parameter, associated with the result of a measurement (eg a

calibration or test) that defines the range of the values that could
reasonably be attributed to the measured quantity. When uncertainty is
evaluated and reported in a specified way it indicates the level of
confidence that the value actually lies within the range defined by the
uncertainty interval.
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Laboratory Quality b) Any measurement is subject to imperfections; some of these are due to
Management System
random effects, such as short-term fluctuations in temperature,
humidity and air-pressure or variability in the performance of the
measurer. Repeated measurements will show variation because of these
random effects. Other imperfections are due to the practical limits to
which correction can be made for systematic effects, such as offset of a
measuring instrument, drift in its characteristics between calibrations,
personal bias in reading an analogue scale or the uncertainty of the
value of a reference standard.

4) ● Selectivity/specificity and Confirmation of identity,

• Limit of detection,
• Limit of quantification,
• Working and linear ranges,
• Recovery (Accuracy/trueness),
• Precision (Repeatability and Reproducibility), and
• Robustness and Ruggedness.

15.15 SUGGESTED READING

Accreditation for Chemical Laboratories, UKAS Publication ref: LAB 27,
Edition 1, 2000.

CITAC- EURACHEM Guide to Quality in Analytical Chemistry - An Aid to

Accreditation, Edition 2002.

EEE/ RM/ 062rev3 The Selection and use of Reference Materials - A Basic
Guide for Laboratories and Accreditation Bodies. A EA-EURACHEM
publication.

EURACHEM Guide “The Fitness for Purpose of Analytical Methods - A

Laboratory Guide to Method Validation and Related Topics”, First Edition,
1998

EURACHEM/CITAC Guide CG 4 “Quantifying Uncertainty in Analytical

Measurement”, Second Edition, 2000.

Guide 43 Part 2:1997 Proficiency Testing by Inter Laboratory Comparisons -

Part 2: Selection and use of Proficiency Testing Schemes by Laboratory
Accreditation Bodies.

ILAC-G12:2000 Guidelines for the Requirements for the Competence of

Reference Material Producers.

ISO Guide 30:1992 Terms and Definitions used in Connection with Reference
Materials; ISO Guide 31:2000 Contents of Certificates of Reference
Materials; ISO Guide 32:1997 Calibration of Chemical Analysis and use of
Certified Reference Materials; ISO Guide 33 :2000 Use of Certified Reference
Materials;

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ISO Guide 43 Part 1:1997 Proficiency Testing by Inter Laboratory General Topics:
Comparisons – Part 1: Development and Operation of Proficiency Testing Related to Food
Testing Laboratories
Schemes and ISO.

ISO Guide 34:2000 Quality System Guidelines for the Production of Reference
Materials;

ISO Guide 35:2006 Certification of Reference Materials - General and

Statistical Principles.

NABL 103 - Specific Guidelines for Chemical Testing Laboratories.

Technical note: c&b-002, February 2005 “Quality Assurance of Equipment

Commonly used in Chemical and Biological Testing Laboratories”, Issued by
Singapore accreditation council - Singapore laboratory accreditation scheme
(sac-singlas).

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