SIST-TS CEN/TS 17455:2020

SLOVENSKI STANDARD
kSIST-TS FprCEN/TS 17455:2019
01-december-2019
Krma - Metode vzorčenja in analize - Izvedbena merila v posameznem laboratoriju
in v primerjalnem preskusu validirane metode analiz za določanje mikotoksinov
Animal feeding stuffs - Methods of sampling and analysis - Performance criteria for
single laboratory validated and ring-trial validated methods of analysis for the
determination of mycotoxins
Kriterienansatz für Methoden zur Analyse von Mycotoxinen in Futtermitteln
Aliments des animaux - Méthodes d’échantillonnage et d’analyse - Critères de
performance des méthodes d’analyse des mycotoxines validées dans un seul laboratoire
ou suite à un essai interlaboratoires
Ta slovenski standard je istoveten z: FprCEN/TS 17455
ICS:
65.120 Krmila Animal feeding stuffs
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
kSIST-TS FprCEN/TS 17455:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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kSIST-TS FprCEN/TS 17455:2019


FINAL DRAFT
TECHNICAL SPECIFICATION
FprCEN/TS 17455
SPÉCIFICATION TECHNIQUE

TECHNISCHE SPEZIFIKATION

October 2019
ICS 65.120; 71.040; 71.040.50
English Version

Animal feeding stuffs - Methods of sampling and analysis -
Performance criteria for single laboratory validated and
ring-trial validated methods of analysis for the
determination of mycotoxins
Aliments des animaux - Méthodes d'échantillonnage et Kriterienansatz für Methoden zur Analyse von
d'analyse - Critères de performance des méthodes Mycotoxinen in Futtermitteln
d'analyse des mycotoxines validées dans un seul
laboratoire ou suite à un essai interlaboratoires


This draft Technical Specification is submitted to CEN members for Vote. It has been drawn up by the Technical Committee
CEN/TC 327.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
United Kingdom.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.

Warning : This document is not a Technical Specification. It is distributed for review and comments. It is subject to change
without notice and shall not be referred to as a Technical Specification.


EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. FprCEN/TS 17455:2019 E
worldwide for CEN national Members.

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Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Relation to other technical document definitions or concepts . 14
4.1 Reporting of sum parameters . 14
4.2 Requirements for multi-mycotoxin methods . 14
4.3 Identification criteria . 14
4.4 Limit of quantification (determination) and limit of detection . 14
5 Performance Criteria for analytical methods for mycotoxins . 15
5.1 General Criteria for methods of analysis for mycotoxins . 15
5.1.1 Overall concepts . 15
5.1.2 Screening methods for mycotoxins . 15
5.1.3 Confirmatory methods for mycotoxins . 15
5.2 Performance characteristics . 16
5.2.1 General . 16
5.2.2 Recovery . 16
5.2.3 RSD . 17
R
5.2.4 RSD . 20
r
5.2.5 RSD . 20
Ri
5.2.6 Limit of quantification . 21
5.2.7 Minimum applicability range . 21
Bibliography . 23

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European foreword
This document (FprCEN/TS 17455:2019) has been prepared by Technical Committee CEN/TC 327
“Animal feeding stuffs - Methods of sampling and analysis”, the secretariat of which is held by NEN.
This document is currently submitted to the Formal Vote.
This document has been prepared under a standardization request given to CEN by the European
Commission and the European Free Trade Association.
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Introduction
The European Committee for Standardization (CEN) selects and elaborates methods of analysis for
organic contaminants that are to become European Standards. These standards can be used for those
contaminants that are subject to regulation. When used for this purpose, the main functions of a
standard is to enable feed manufacturers to determine with reasonable certainty whether a
consignment may be put on the market and to enable regulatory authorities to determine equitably
whether feedstuffs on the market comply with legal or recommended limits.
CEN/TC 327/working group (WG) 5 decided to establish a criteria guide in order to allow
benchmarking of methods of analysis for their fitness for purpose [1]. The performance criteria laid
down therein are based on published data, collected from official reports on inter-laboratory studies
[2]-[12].
Where performance characteristics are absent or limited in availability, the criteria were estimated
based on the experiences and opinions of the experts of the CEN working group. The selection criteria
could need updating in future revisions of this document, if newer or more accurate data on method
performance characteristics become available. This document lists relevant performance parameters
and gives information on their definition. It further describes how they can be practically obtained; it
indicates guidance values demonstrating fitness for purpose. As a result, these guidance values can be
expected from (derived by) experienced analytical laboratories.
This document may contain useful information for CEN members, the European Commission, the
EFTA secretariat, other governmental agencies or outside bodies. The criteria in this CEN report are
used as guidance in the CEN/TC 327/WG 5. In general, method performance criteria are generated
for collaborative trials or for single laboratory validation studies. The first case describes how a
method performs when used by several laboratories, giving greater confidence that the method is
applicable and can be implemented with the obtained method performance. The second case only
demonstrates how a method performs in a particular laboratory. As a result, single laboratory
validation should be conducted with care to avoid misinterpretation. This is important as
reproducibility (between laboratory variance) can only be derived from collaborative trials.
Furthermore, performance parameters such as limit of detection (LOD) and limit of quantification
(LOQ) are laboratory specific as the quantitative measurement capability strongly depends on the
instrumental conditions used by each laboratory.
This document relates to current EU legislation concerning performance requirements of analytical
methods for mycotoxins in food and feed chain, such as Regulation (EC) No 2017/625 on official
controls performed to ensure the verification of compliance with feed and food law, animal health
and animal welfare rules [13] as well as Regulation (EC) No 401/2006 laying down the methods of
sampling and analysis for the official control of the levels of mycotoxins in foodstuffs [14]. The latter
is specific for food, however as recent legislation on some mycotoxins does not specify the intended
final use, either food or feed, rather the raw commodity (Commission Recommendation
2013/165/EU on the presence of T-2 and HT-2 toxin in cereals and cereal products) performance
requirements for analytical methods are in the future anticipated to be equivalent independent of the
commodities' destination purpose.
Further this criteria approach document also relates to pre-existing standards such as the ISO 5725-
series, CR 13505 [15] and CEN/TR 16059 [16].
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1 Scope
This document specifies performance criteria for the selection of single-laboratory validated or
collaborative study validated methods of analysis of mycotoxins in feed. The terms and definition of
the relevant parameters for method validation are included. The performance requirements and
characteristics are provided. This document could serve as a guide:
— to assess the quality of new European Standard methods under validation;
— to review the quality of previous collaborative trials;
— to confirm the extension of the scope of an already published European Standard applied to
other analyte concentrations or matrices; or
— to evaluate the fitness-for-purpose of single-validated methods.
The performance criteria can apply to methods dedicated to the determination of mycotoxins.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at http://www.iso.org/obp
3.1
accuracy
closeness of agreement between a test result and the accepted reference value
Note 1 to entry: 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.
Note 2 to entry: It is assessed by determining trueness and precision.
[SOURCE: ISO 5725-1:1994, 3.6, see [17], modified; 2002/657/EEC, see [18]modified]
3.2
applicability
scope of the analytical method; description of the analytes, matrices, and concentration ranges (mass
fractions) for which a method of analysis can be used satisfactorily to determine compliance with a
given standard (i.e. CEN, ISO, CODEX)
Note 1 to entry: In addition to a statement of the range of capability of satisfactory performance for each
factor, the statement of applicability (scope) also includes warnings as to known interference by other analytes,
or inapplicability to certain matrices and situations.
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3.3
bias
difference between the expectation of the test results (x) and an accepted reference value (x )
ref
Note 1 to entry: The bias can be expressed in absolute or relative terms (b or b(%), respectively) as:
b xx−
 (1)
ref
xx−
ref
b % ×100 (2)
( )
x
ref
[SOURCE: ISO 5725-1:1994, 3.8, see [17], modified; Eurachem 2014, see [19], modified]
3.4
inter-laboratory comparison
organisation, performance and evaluation of measurements or tests on the same or similar items by
two or more laboratories in accordance with predetermined conditions
Note 1 to entry: These conditions include that the materials have a suitable homogeneity and acceptable
stability that is documented.
Note 2 to entry: The larger the number of participating laboratories, the greater the confidence that can be
placed in the resulting statistical parameters.
Note 3 to entry: Vague terms such as “round-robins”, “intercalibrations”, “ring tests”, etc. should not be used.
Studies involving several laboratories with each preparing its own test materials and using its own methods,
are sometimes called cooperative studies.
[SOURCE: ISO/IEC 17043:2010, 3.4, see [20], modified]
3.4.1
collaborative study
inter-laboratory comparison aiming at the evaluation of the performance characteristics of a method
Note 1 to entry It means analysing the same sample by the same method to determine the performance
characteristics of the method.
Note 2 to entry The study covers random measurement error and laboratory bias.
Note 3 to entry: The reported results are used to estimate the performance characteristics of the method,
such as precision parameters like repeatability (within laboratory) and reproducibility (between laboratories).
When necessary and possible, other pertinent characteristics such as systematic error and/or recovery can be
reported. These can include bias, recovery, internal quality control parameters, sensitivity, limit of
quantification, and applicability.
[SOURCE: 2002/657/EEC, see [18], modified]
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3.4.2
proficiency testing
evaluation of participant performance against pre-established criteria by means of inter-laboratory
comparisons
Note 1 to entry: In practise the study consists of one or more analyses or measurements by a group of
laboratories on one or more homogeneous, stable test samples by the method selected or used by each
laboratory.
Note 2 to entry: The reported results are compared with those from other laboratories or with the known or
assigned reference value, usually with the objective of evaluating or improving laboratory performance.
Note 3 to entry Proficiency testing means analysing the same sample allowing laboratories to choose their
own methods, provided these methods are used under routine conditions.
[SOURCE: 2002/657/EEC, see [18], modified; ISO/IEC 17043:2010, 3.7, see [20], modified]
3.4.3
reference material characterisation
inter-laboratory comparison aiming at the assignment of values to reference materials and
assessment of their suitability for use in specific test or measurement procedures
Note 1 to entry: The assigned values are usually the reference value (best estimate of the “true value”) to a
quantity (concentration/mass fraction) in the test material, usually with a stated uncertainty.
3.5
limit of detection
x
LOD
minimum amount or concentration of the analyte in a test sample which can be detected reliably but
not necessarily quantified, as demonstrated by a collaborative trial or other appropriate validation
Note 1 to entry: Guidelines for the sound generation of LOD values can be found in [23].
Note 2 to entry: For analytical systems where the validation range does not include or approach it, the
detection limit does not need to be part of a validation.
Note 3 to entry: The various conceptual approaches to the subject depend on the estimate of precision at or
near zero concentration (mass fraction) under repeatability or reproducibility conditions.
[SOURCE: Thompson et al. 2002, see [21], modified; ISO 24276:2006, 3.1.6, see [22], modified]
3.6
limit of quantification
x
LOQ
lowest concentration or amount of the analyte in a test sample which can be quantitatively
determined with an acceptable level of precision and accuracy, as demonstrated by a collaborative
trial or other appropriate validation
Note 1 to entry: Rules for the sound generation of LOQ values can be found in [23].
Note 2 to entry: Measurements below LOQ are not void of information content. However, they are not of use
for method criteria assessment
[SOURCE: Thompson et al. 2002, see [21], modified; ISO 24276:2006, 3.1.7, see [22], modified]
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3.7
linearity
ability (within a given range) of an analytical procedure to obtain test results which are directly
proportional to the concentration (mass fraction) of analyte in the sample
[SOURCE: VAM/III/5626/94, see [24]]
3.8
lowest validated level
lowest concentration (mass fraction) investigated in the frame of a collaborative study or a single-
laboratory validation
3.9
precision
closeness of agreement between independent test results obtained under stipulated conditions
Note 1 to entry: Precision depends only on the distribution of random errors and does not relate to the true
value, conventional true value or specified value.
Note 2 to entry: The measure of precision is usually expressed in terms of imprecision and computed as a
standard deviation of the test results. Less precision is reflected by a larger standard deviation.
Note 3 to entry: “Independent test results” means results obtained in a manner not influenced by any
previous result on the same or similar test object. Quantitative measure of precision depends critically on the
stipulated conditions. Repeatability and reproducibility conditions are particular sets of extreme conditions.
[SOURCE: ISO 5725-1:1994, 3.12, see [17]]
3.9.1
repeatability
RSD
r
precision under repeatability conditions
[SOURCE: ISO 5725-1:1994, 3.13, see [17]]
3.9.1.1
repeatability conditions
conditions where independent test results are obtained with the same method on identical test items
in the same laboratory by the same operator using the same equipment within short intervals of time
[SOURCE: ISO 5725-1:1994, 3.14, see [17]]
3.9.1.2
repeatability standard deviation
s
r
standard deviation of test results obtained under repeatability conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test results under repeatability
conditions.
Note 2 to entry: Similarly “repeatability variance” and “repeatability coefficient of variation” could be defined
and used as measures of the dispersion of the test results under repeatability conditions.
[SOURCE: ISO 5725-1:1994, 3.15, see [17]]
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3.9.1.3
repeatability relative standard deviation
RSD
r
relative standard deviation of test results obtained under repeatability conditions
Note 1 to entry: The repeatability relative standard deviation (RSD ) can be expressed as follows:
r
s
r
RSD (%) × 100 (4)
r
c
mean
where:
c is the mean concentration (mass fraction).
mean
3.9.1.4
repeatability limit
r
value less than or equal to which the absolute difference between two test results obtained under
repeatability conditions may be expected to be within a probability of 95 %
Note 1 to entry: The repeatability limit (r) can be expressed as follows:
rs22× (5)
r
[SOURCE: ISO 5725-1:1994, 3.16, see [17]]
3.9.2
intermediate precision
precision under within-laboratory reproducibility conditions
3.9.2.1
within-laboratory reproducibility conditions
conditions where test results are obtained with the same method on identical test items on different
days with preferably different operators using different equipment
3.9.2.2
within-laboratory reproducibility standard deviation
s
Ri
standard deviation of test results obtained under within-laboratory reproducibility conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test results under within-laboratory
reproducibility conditions
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3.9.2.3
within-laboratory reproducibility relative standard deviation
RSD
Ri
relative standard deviation of test results obtained under within-laboratory reproducibility
conditions
Note 1 to entry: The within-laboratory reproducibility relative standard deviation (RSD ) can be expressed
Ri
as follows:
s
Ri
RSD % × 100
(6)
( )
Ri
c
mean
where
s is the standard deviation, calculated from results generated under within-laboratory
Ri
reproducibility conditions;
c is the mean concentration (mass fraction).
mean
3.9.3
reproducibility
precision under reproducibility conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test results under reproducibility
conditions.
Note 2 to entry: Equivalent terms used are “reproducibility variance” and “reproducibility coefficient of
variation” but should not be used in this context.
[SOURCE: ISO 5725-1:1994, 3.17, see [17], modified]
3.9.3.1
reproducibility conditions
conditions where test results are obtained with the same method on identical test items in different
laboratories with different operators using different equipment
[SOURCE: ISO 5725-1:1994, 3.18, see [17]]
3.9.3.2
reproducibility standard deviation
s
R
standard deviation of test results obtained under reproducibility conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test results under reproducibility
conditions.
Note 2 to entry: Similarly “reproducibility variance” and “reproducibility coefficient of variation” could be
defined and used as measures of the dispersion of the test results under reproducibility conditions.
[SOURCE: ISO 5725-1:1994, 3.19, see [17]]
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3.9.3.3
reproducibility relative standard deviation
RSD
R
relative standard deviation, calculated from results generated under reproducibility conditions
Note 1 to entry: The reproducibility relative standard deviation (RSD ) can be expressed as follows:
R
s
R
RSD (%) × 100 (7)
R
c
mean
where
S is the standard deviation, calculated from results generated under reproducibility conditions
R
c is the mean concentration (mass fraction).
mean
3.9.3.4
reproducibility limit
R
value less than or equal to which the absolute difference between two test results obtained under
reproducibility conditions may be expected to be within a probability of 95 %
Note 1 to entry: The reproducibility limit (R) can be expressed as follows:
R 22× s
(8)
R
[SOURCE: ISO 5725-1:1994, 3.20, see [17], modifed]
3.10
recovery
R

rec
percentage of the true concentration of a substance recovered during the analytical procedure
Note 1 to entry: An ideal approach to demonstrate that recovery is consistent over the whole working range
is to conduct a series at least 4 fortification experiments over the whole working range. Linearity shall be
demonstrated by a “lack of fit” test. The slope of the curve for recovery determination ( × 100 for expression in
%), provided the fortified increment mass fraction (c ) is plotted as X-axis against the measured mass fraction
+
(c - c ) on the Y-axis.
f 0
Note 2 to entry: Practical information for considerations in setting up recovery experiments are given in [25].
Note 3 to entry: The relative recovery (R ) can be expressed as:
rec
x
R (%) × 100
(9)
rec
x
ref
where:
x is the measured concentration (mass fraction);
x is the reference concentration (mass fraction)
ref
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Note 4 to entry: For fortified (spiked) samples, the relative recovery (R ) is the difference observed after
rec
analysis of the test material and fortified (spiked) to the test material and can be expressed as:
xx−
f0
R (%) × 100 (10)
rec
x
spike
where:
x is the measured concentration (mass fraction) in the spiked or fortified sample;
f
x is the measured concentration (mass fraction) in the unfortified sample;
0
x is the spiked/added concentration (mass fraction).
spike
Note 5 to entry: Bias and recovery are correlated as follows: Recovery (%) + bias (%) = 100 %.
Note 6 to entry: The amount added for recovery estimation should be a substantial fraction of, or more than,
the amount present in the unfortified material. Ideally, the unfortified material should contain no measurable
level of the analyte under test.
Note 7 to entry: A true or assigned value is known only in cases of fortified materials, certified reference
materials, or by analysis by another (presumably unbiased) method. The concentration (mass fraction) in the
unfortified material is obtained by direct analysis or by the method of standard additions. In other cases, there
is no direct measure of bias, and consensus values derived from the collaborative study itself often can be used
for the reference point.
[SOURCE: EuraChem Guide, see [19], modified]
3.11
ruggedness
resistance to changes in the results produced by an analytical method when minor deviations are
made from the experimental conditions described in the procedure
Note 1 to entry: It is the susceptibility of an analytical method to changes in experimental conditions which
can be expressed as a list of the sample materials, analytes, storage conditions, environmental and/or sample
preparation conditions under which the method can be applied as presented or with specified minor
modifications that are likely to occur during the analytical procedure.
Note 2 to entry: For all experimental conditions which could in practice be subject to fluctuation (e.g.
stability of reagents, composition of the sample, pH, temperature) any variations which could affect the
analytical result should be indicated.
Note 3 to entry: The term robustness is not recommended [24], as ruggedness is already been used by
Thomason et al. [21] and additional wording could lead to confusion.
[SOURCE: Thompson et al. 2002, see [21], modified; 2002/657/EEC, see [18], modified]
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3.12
selectivity
degree to which a method can quantify the analyte accurately in the presence of interferences
Note 1 to entry: For the determination and identification of mycotoxin the interferences are usually matrix
(feed material) derived components of similar physico-chemical behaviour, either exhibiting similar properties
during detection and/or separation such as sample clean-up or chromatographic separation.
Note 2 to entry: IUPAC recommends the use of the term “selectivity” and discourages the use of the term
“specificity”. IUPAC clarified this overlap by expressing the view that “Specificity is the ultimate of Selectivity”
Note 3 to entry: Ideally, selectivity should be evaluated for any important interferences likely to be present.
It is particularly important to check interferences that are likely, on physico-chemical principles, to respond to
the test. As a general principle, selectivity should be sufficiently good for any interferences to be ignored.
[SOURCE: Thompson et al. 2002, see [21], modified; Wessmann et al. 2001, see [26], modifed]
3.13
sensitivity
gradient or slope of the calibration function of a method
Note 1 to entry: As this is usually arbitrary, depending on instrumental settings, it is not useful in validation.
It could be useful in quality assurance procedures, however, to test whether an instrument is performing to a
co
...