Animal feeding stuffs: Methods of sampling and analysis - Determination of pentachlorophenol (PCP) in feed materials and compound feed by LC-MS/MS

This document specifies a liquid chromatographic method with triple-quadrupole mass spectrometry (MS/MS) detection for the determination of pentachlorophenol (PCP) in feed materials and animal feed.
The limit of quantitation (LOQ) for the PCP determination in guar gum, fatty acid distillates (FAD) and animal feed is 10 µg/kg. Individual laboratories are responsible for ensuring that the equipment that they use will achieve this limit of quantification.
The method is validated in an international collaborative trial for pentachlorophenol in compound feed, guar gum and fatty acid distillate in the range between 9 µg/kg and 22 µg/kg.
The results of the collaborative trial, in which 16 laboratories participated, have shown that the method is applicable for the determination of PCP in compound feed, guar gum and FAD at the desired limit of 10 µg/kg. Satisfactory results were obtained for one compound feed sample, guar gum and the two FAD samples (HorRat <2), while for the second compound feed sample a HorRat value of 2,2 was obtained.

Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von Pentachlorphenol (PCP) in Futtermittel und Mischfuttermittel mittels LC-MS/MS

Dieses Dokument legt ein flüssigkeitschromatographisches Verfahren mit Nachweis mittels Triple-Quadrupol Massenspektrometrie (MS/MS) für die Bestimmung von Pentachlorphenol (PCP) in Einzelfutter  und Mischfuttermitteln fest.
Die Bestimmungsgrenze (LOQ) für die PCP Bestimmung in Guargummi, Fettsäuredestillaten (FAD) und Futtermitteln liegt bei 10 µg/kg. Die einzelnen Laboratorien sind selbst dafür verantwortlich sicherzustellen, dass die von ihm verwendeten Geräte diese Bestimmungsgrenze auch erreichen.
Das Verfahren ist in einem internationalen Ringversuch zur Pentachlorphenol Bestimmung in Mischfutter, Guargummi und Fettsäuredestillat im Bereich zwischen 9 µg/kg und 22 µg/kg validiert worden.
Die Ergebnisse des Ringversuchs, an dem 16 Laboratorien teilnahmen, haben gezeigt, dass das Verfahren für die Bestimmung von PCP in Mischfutter, Guargummi und FAD beim gewünschten Grenzwert von 10 µg/kg anwendbar ist. Zufriedenstellende Ergebnisse (HorRat Wert < 2) wurden für eine Mischfutter Probe, Guargummi und die beiden FAD Proben erhalten, während bei der zweiten Mischfutter Probe ein HorRat Wert von 2,2 erhalten wurde.

Aliments pour animaux : Méthodes d’échantillonnage et d’analyse - Détermination de la teneur en pentachlorophénol (PCP) dans les matières premières pour aliments des animaux et les aliments composés pour animaux par CL-SM/SM

Le présent document spécifie une méthode de chromatographie liquide couplée à la détection par spectrométrie de masse triple quadripôle (SM/SM) pour la détermination de la teneur en pentachlorophénol (PCP) dans les matières premières pour aliments des animaux et les aliments pour animaux.
La limite de quantification (LOQ) applicable à la détermination de la teneur en PCP dans la gomme de guar, les distillats d’acides gras (FAD) et les aliments pour animaux est de 10 µg/kg. Chaque laboratoire est tenu de s’assurer que l’équipement qu’il utilise atteindra cette limite de quantification.
La méthode a été validée lors d’un essai interlaboratoires international portant sur le pentachlorophénol présent dans les aliments composés pour animaux, la gomme de guar et les distillats d’acides gras dans la gamme allant de 9 µg/kg à 22 µg/kg.
Les résultats de l’essai interlaboratoires, auquel ont participé 16 laboratoires, ont montré que la méthode est applicable à la détermination de la teneur en PCP dans les aliments composés pour animaux, la gomme de guar et les FAD à la limite souhaitée de 10 µg/kg. Des résultats satisfaisants ont été obtenus pour un échantillon d’aliment composé, la gomme de guar et les deux échantillons de FAD (HorRat < 2), alors qu’une valeur de HorRat de 2,2 a été obtenue pour le deuxième échantillon d’aliment composé.

Krma: metode vzorčenja in analize - Določevanje pentaklorofenola (PCP) v sestavinah krme in krmni mešanici z LC-MS/MS

General Information

Status
Published
Public Enquiry End Date
24-Apr-2019
Publication Date
18-Jun-2020
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
19-May-2020
Due Date
24-Jul-2020
Completion Date
19-Jun-2020

Buy Standard

Standard
EN 17362:2020
English language
19 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day
Draft
prEN 17362:2019
English language
18 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN 17362:2020
01-julij-2020
Krma: metode vzorčenja in analize - Določevanje pentaklorofenola (PCP) v
sestavinah krme in krmni mešanici z LC-MS/MS
Animal feeding stuffs: Methods of sampling and analysis - Determination of
pentachlorophenol (PCP) in feed materials and compound feed by LC-MS/MS
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von
Pentachlorphenol (PCP) in Futtermittel und Mischfuttermittel mittels LC-MS/MS
Aliments pour animaux : Méthodes d’échantillonnage et d’analyse - Détermination de la
teneur en pentachlorophénol (PCP) dans les matières premières pour aliments des
animaux et les aliments composés pour animaux par CL-SM/SM
Ta slovenski standard je istoveten z: EN 17362:2020
ICS:
65.120 Krmila Animal feeding stuffs
SIST EN 17362:2020 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST EN 17362:2020

---------------------- Page: 2 ----------------------
SIST EN 17362:2020


EN 17362
EUROPEAN STANDARD

NORME EUROPÉENNE

April 2020
EUROPÄISCHE NORM
ICS 65.120
English Version

Animal feeding stuffs: Methods of sampling and analysis -
Determination of pentachlorophenol (PCP) in feed
materials and compound feed by LC-MS/MS
Aliments pour animaux : Méthodes d'échantillonnage Futtermittel: Probenahme- und
et d'analyse - Détermination de la teneur en Untersuchungsverfahren - Bestimmung von
pentachlorophénol (PCP) dans les matières premières Pentachlorphenol (PCP) in Futtermittel und
pour aliments des animaux et les aliments composés Mischfuttermittel mittels LC-MS/MS
pour animaux par CL-SM/SM
This European Standard was approved by CEN on 7 March 2020.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.

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.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17362:2020 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
Contents Page
European foreword . 3
1 Scope . 4
2 Normative references . 4
3 Terms and definitions . 4
4 Principle . 7
5 Reagents and materials . 7
5.1 General . 7
5.2 Chemicals . 7
6 Apparatus . 10
6.1 Analytical balance, accuracy to 0,1 mg . 10
6.2 Centrifuge . 10
6.3 Water bath . 10
6.4 Liquid chromatograph connected to a triple-quadrupole mass spectrometer . 10
7 Sampling . 11
8 Preparation of the test sample . 12
9 Procedure. 12
9.1 General . 12
9.2 Test portions and extraction . 12
9.3 LC-MS/MS . 13
10 Calculation and expression of results . 13
10.1 General . 13
10.2 Calibration criteria . 14
10.3 Identification and confirmation. 14
10.4 Calculation . 14
10.5 Recovery . 15
11 Precision . 15
11.1 Collaborative test . 15
11.2 Repeatability limit . 15
11.3 Reproducibility limit . 15
12 Test report . 16
Annex A (informative) Collaborative trial. 17
Bibliography . 19

2

---------------------- Page: 4 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
European foreword
This document (EN 17362:2020) 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 shall be given the status of a national standard, either by publication of an identical text
or by endorsement, at the latest by October 2020, and conflicting national standards shall be withdrawn
at the latest by October 2020.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document has been prepared under a standardization request given to CEN by the European
Commission and the European Free Trade Association.
According to the CEN-CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to implement this document: 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 the United
Kingdom.
3

---------------------- Page: 5 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
1 Scope
This document specifies a liquid chromatographic method with triple-quadrupole mass spectrometry
(MS/MS) detection for the determination of pentachlorophenol (PCP) in feed materials and compound
feed.
The limit of quantitation (LOQ) for the PCP determination in guar gum, fatty acid distillates (FAD) and
compound feed is 10 µg/kg. Individual laboratories are responsible for ensuring that the equipment that
they use will achieve this limit of quantification.
The method is validated in an international collaborative trial for pentachlorophenol in compound feed,
guar gum and fatty acid distillate in the range between 9 µg/kg and 22 µg/kg.
The results of the collaborative trial, in which 16 laboratories participated, have shown that the method
is applicable for the determination of PCP in compound feed, guar gum and FAD at the desired limit of
10 µg/kg. Satisfactory results were obtained for one compound feed sample, guar gum and the two FAD
samples (HorRat < 2), while for the second compound feed sample a HorRat value of 2,2 was obtained.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 6498, Animal feeding stuffs — Guidelines for sample preparation (ISO 6498)
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 https://www.iso.org/obp
3.1
calibration
complete set of operations which estimates under specified conditions the calibration function from
observations of the response variable obtained on reference states
[SOURCE: ISO 3534-2:2006, 3.5.13 [1]]
3.2
collaborative trial
interlaboratory comparisons
organization, performance and evaluation of measurements or tests on the same or similar items by two
or more laboratories in accordance with predetermined conditions
[SOURCE: EN ISO/IEC 17043:2010, 3.4 [2]]
4

---------------------- Page: 6 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
3.3
feed material
products of vegetable or animal origin, whose principal purpose is to meet animals’ nutritional needs, in
their natural state, fresh or preserved, and products derived from the industrial processing thereof, and
organic or inorganic substances, whether or not containing feed additives, which are intended for use in
oral animal-feeding either directly as such, or after processing, or in the preparation of compound feed,
or as carrier of premixtures
[SOURCE: Regulation (EC) No 767/2009 (Article 3(2)(g)) [3]]
3.4
HorRat
ratio of the reproducibility relative standard deviation to that calculated from the Horwitz equation
–0,15
Note 1 to entry: Predicted relative standard deviation PRSD = 2 C
R
HorRatR = RSDR / PRSDR (1)
HorRatr = RSDr / PRSDr (2)
Note 2 to entry: If applied to within-laboratory studies, the normal range of HorRat(r) is 0,30 to 1,30.
−6
Note 3 to entry: To check proper calculation of PRSDR, a C of 10 should give a PRSDR of 16 %. C is concentration
expressed as a mass fraction (both numerator and denominator expressed in the same units). The HorRat is
indicative of method performance for a large majority of methods in chemistry. Normal values lie between 0,50 and
2,00.
[SOURCE: ISO 16577:2016, 3.75 [4]]
3.5
limit of quantitation
measured quantity value, obtained by a given measurement procedure, which is the lowest concentration
of a measurand that can be determined with an acceptable level of repeatability precision and trueness
Note 1 to entry: The “limit of quantitation” is not a concept defined in ISO/IEC Guide 99:2007 [5] but has been
defined as described above.
Note 2 to entry: The abbreviation LOQ is sometimes used.
3.6
precision
closeness of agreement between indications or measured quantity values obtained by replicate
measurements on the same or similar objects under specified conditions
Note 1 to entry: Measurement precision is usually expressed numerically by measures of imprecision, such as
standard deviation, variance, or coefficient of variation under the specified conditions of measurement.
Note 2 to entry: The ‘specified conditions’ can be, for example, repeatability conditions of measurement,
intermediate precision conditions of measurement, or reproducibility conditions of measurement (see ISO 5725-1
[6]).
Note 3 to entry: Measurement precision is used to define measurement repeatability, intermediate measurement
precision, and measurement reproducibility.
Note 4 to entry: Sometimes 'measurement precision' is erroneously used to mean 'measurement accuracy'.
5

---------------------- Page: 7 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
[SOURCE: ISO IEC Guide 99:2007, 2.15 [5]]
3.7
repeatability
precision under repeatability conditions
Note 1 to entry: Repeatability can be expressed quantitatively in terms of the dispersion characteristics of the
results.
[SOURCE: ISO 3534-2:2006, 3.3.5 [1]]
3.8
repeatability limit
r
repeatability critical difference for a specified probability of 95 %
[SOURCE: ISO 3534-2:2006, 3.3.9 [1]]
3.9
repeatability standard deviation
standard deviation of test results or measurement results obtained under repeatability conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test or measurement results under
repeatability conditions.
Note 2 to entry: Similarly, “repeatability variance” and “repeatability coefficient of variation” can be defined and
used as measures of the dispersion of test or measurement results under repeatability conditions.
[SOURCE: ISO 3534-2:2006, 3.3.7 [1]]
3.10
reproducibility
precision under reproducibility conditions
Note 1 to entry: Reproducibility can be expressed quantitatively in terms of the dispersion characteristics of the
results.
Note 2 to entry: Results are usually understood to be corrected results.
[SOURCE: ISO 3534-2:2006, 3.3.10 [1]]
3.11
reproducibility limit
R
reproducibility critical difference for a specified probability of 95 %
[SOURCE: ISO 3534-2:2006, 3.3.14 [1]]
6

---------------------- Page: 8 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
3.12
reproducibility standard deviation
standard deviation of test results or measurement results obtained under reproducibility conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test or measurement results under
reproducibility conditions.
Note 2 to entry: Similarly, “reproducibility variance” and “reproducibility coefficient of variation” can be defined
and used as measures of the dispersion of test or measurement results under reproducibility conditions.
[SOURCE: ISO 3534-2:2006, 3.3.12 [1]]
4 Principle
In order to check for the presence of PCP, a test portion of sample material is fortified with internal
13
standards ( C-PCP). The test portion is extracted using a QuEChERS approach for compound feed and
FAD.
A ‘reversed’ QuEChERS approach is used for guar gum. For FAD samples, lipids are removed prior to
QuEChERS extraction by the addition of sulfuric acid.
Final extracts from all matrices are analysed by liquid chromatography coupled to triple quadrupole mass
spectrometry, operated in negative electrospray ionization mode.
Identification is done on the basis of retention time and mass-to-charge ratio. Quantification is done using
the internal standard method.
5 Reagents and materials
5.1 General
Use only reagents of recognized analytical grade and with a purity suitable for organochlorine residue
analysis. Check the purity of the reagents by performing a blank test under the same conditions as used
in the method. The chromatogram should not show any interfering impurity at the retention time of
compounds of interest.
5.2 Chemicals
5.2.1 Diethylamine (DEA)
5.2.2 Acetonitrile (ACN)
5.2.3 Methanol
5.2.4 Acetone
5.2.5 n-Hexane
5.2.6 Sulphuric acid (concentrated, 95 % – 98 %)
5.2.7 Deionized water
7

---------------------- Page: 9 ----------------------
SIST EN 17362:2020
EN 17362:2020 (E)
5.2.8 Sodium hydroxide
5.2.8.1 Sodium hydroxide solution (10 M)
Weigh 4 g of sodium hydroxide (5.2.8) and add 10 ml deionized water (5.2.7) to achieve a concentration
of 10 M. Store at room temperature in a closed glass bottle with a plastic cap or stopper.
5.2.9 Magnesium sulphate
5.2.10 Sodium chloride
5.2.11 Formic acid
5.2.12 Ammonium formate
5.2.12.1 Ammonium formate solution (1 M)
Weigh 6,3 g ammonium formate (5.2.12) and add 100 ml deionized water (5.2.7) to achieve a
concentration of 1 M. Store the solution at room temperature. The solution is stable under these
conditions for at least 1 month.
5.2.13 Mobile phase A
Take 1 ml ammonium formate solution (1 M) (5.2.12.1) and add 999 ml deionized water (5.2.7) and 20 µL
formic acid (5.2.11). Mix well. Store the solution at room temperature. The solution is stable under these
conditions for at least 1 month.
5.2.14 Mobile phase B
Take 1 ml ammonium formate solution (1 M) (5.2.12.1) and add 50 ml deionized water (5.2.7), 949 ml
methanol (5.2.3) and 20 µl formic acid (5.2.11) and mix well. Store the solution at room temperature. The
solution is stable under these conditions for at least 1 month.
5.2.15 Pentachlorophenol (PCP sodium salt, 95 % purity)
5.2.15.1 PCP Stock solution 1 (2 000 µg/ml)
Weigh 25 mg (± 0,01 mg) of PCP (5.2.15) (taking impurities into consideration) and add 12,5 ml
deionized water (5.2.7) to achieve a concentration of 2 000 µg/ml. Store the solution in a refrigerator at
4 °C (± 3 °C). The solution is stable under these conditions for at least 12 months.
5.2.15.2 PCP Stock solution 2 (1 ng/µl)
Dilute 50 µl of PCP Stock solution 1 (5.2.15.1) to 100 ml deionized water (5.2.7) in a volumetric flask
(5.2.20). Store the solutio
...

SLOVENSKI STANDARD
oSIST prEN 17362:2019
01-april-2019
.UPDPHWRGHY]RUþHQMDLQDQDOL]H'RORþHYDQMHSHQWDNORURIHQROD 3&3 Y
VHVWDYLQDKNUPHLQNUPQLPHãDQLFL]/&0606
Animal feeding stuffs: Methods of sampling and analysis - Determination of
pentachlorophenol (PCP) in feed materials and compound feed by LC-MS/MS
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von
Pentachlorphenol (PCP) in Futtermittel und Mischfuttermittel mittels LC-MS/MS
Aliments pour animaux - Méthodes d'échantillonnage et d'analyse - Détermination de la
teneur en pentachlorophénol (PCP) dans les matières premières pour aliments des
animaux et les aliments composés pour animaux par CL-SM/SM
Ta slovenski standard je istoveten z: prEN 17362
ICS:
65.120 Krmila Animal feeding stuffs
oSIST prEN 17362:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
oSIST prEN 17362:2019

---------------------- Page: 2 ----------------------
oSIST prEN 17362:2019


DRAFT
EUROPEAN STANDARD
prEN 17362
NORME EUROPÉENNE

EUROPÄISCHE NORM

February 2019
ICS 65.120
English Version

Animal feeding stuffs: Methods of sampling and analysis -
Determination of pentachlorophenol (PCP) in feed
materials and compound feed by LC-MS/MS

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

If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations
which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.

This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other
language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC
Management Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, 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 European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.


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. prEN 17362:2019 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)
Contents Page
European foreword . 3
1 Scope . 4
2 Normative references . 4
3 Terms and definitions . 4
4 Principle . 7
5 Reagents and materials . 7
5.1 General . 7
5.2 Chemicals . 7
6 Apparatus . 10
7 Sampling . 11
8 Preparation of the test sample . 11
9 Procedure . 11
9.1 General . 11
9.2 Test portions and extraction . 12
9.3 LC-MSMS . 13
10 Calculation and expression of results . 13
10.1 General . 13
10.2 Calibration criteria . 13
10.3 Identification and confirmation . 13
10.4 Calculation . 14
10.5 Recovery . 15
11 Precision . 15
11.1 Collaborative test . 15
11.2 Repeatability limit . 15
11.3 Reproducibility limit . 15
12 Test report . 15
Annex A (informative) Collaborative trial . 16
Bibliography . 18

2

---------------------- Page: 4 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

European foreword
This document (prEN 17362:2019) 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 CEN Enquiry.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
3

---------------------- Page: 5 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

1 Scope
This document specifies a liquid chromatographic method with triple-quadrupole mass spectrometry
(MS/MS) detection for the determination of pentachlorophenol (PCP) in feed materials and animal feed.
The limit of quantitation (LOQ) for the PCP determination in guar gum, fatty acid distillates (FAD) and
animal feed is 10 µg/kg. Individual laboratories are responsible for ensuring that the equipment that
they use will achieve this limit of quantification.
The method is validated in an international collaborative trial for pentachlorophenol in compound feed,
guar gum and fatty acid distillate in the range between 9 µg/kg and 22 µg/kg.
The results of the collaborative trial, in which 16 laboratories participated, have shown that the method
is applicable for the determination of PCP in compound feed, guar gum and FAD at the desired limit of
10 µg/kg. Satisfactory results were obtained for one compound feed sample, guar gum and the two FAD
samples (HorRat <2), while for the second compound feed sample a HorRat value of 2,2 was obtained.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 6498, Animal feeding stuffs — Guidelines for sample preparation (ISO 6498)
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
calibration
complete set of operations which estimates under specified conditions the calibration function from
observations of the response variable obtained on reference states
[SOURCE: ISO 3534-2:2006, 3.5.13]
3.2
collaborative trial
interlaboratory comparisons
organization, performance and evaluation of measurements or tests on the same or similar items by
two or more laboratories in accordance with predetermined conditions
[SOURCE: EN ISO/IEC 17043:2010, 3.4]
4

---------------------- Page: 6 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

3.3
detection limit
limit of detection
measured quantity value, obtained by a given measurement procedure, for which the probability of
falsely claiming the absence of a component in a material is β, given a probability α of falsely claiming
its presence
Note 1 to entry: IUPAC recommends default values for α and β equal to 0,05.
Note 2 to entry: The abbreviation LOD is sometimes used.
Note 3 to entry: The term 'sensitivity' is discouraged for ‘detection limit’.
[SOURCE: ISO/IEC Guide 99:2007, 4.18]
3.4
feed material
products of vegetable or animal origin, whose principal purpose is to meet animals’ nutritional needs,
in their natural state, fresh or preserved, and products derived from the industrial processing thereof,
and organic or inorganic substances, whether or not containing feed additives, which are intended for
use in oral animal-feeding either directly as such, or after processing, or in the preparation of compound
feed, or as carrier of premixtures
[SOURCE: Regulation (EC) No 767/2009 (Article 3, 1g)] [1]
3.5
HorRat
ratio of the reproducibility relative standard deviation to that calculated from the Horwitz equation
–0,15
Note 1 to entry: Predicted relative standard deviation PRSDR = 2 C
HorRat = RSD / PRSD (1)
RR R
HorRat = RSD / PRSD (2)
r r R
Note 2 to entry: If applied to within-laboratory studies, the normal range of HorRat(r) is 0,30 – 1,30.
Note 3 to entry: To check proper calculation of PRSD , a C of 10−6 should give a PRSD of 16 %. C is concentration
R R
expressed as a mass fraction (both numerator and denominator expressed in the same units). The HorRat is
indicative of method performance for a large majority of methods in chemistry. Normal values lie between 0,50
and 2,00.
[SOURCE: ISO 16577:2016, 3.75]
3.6
precision
closeness of agreement between indications or measured quantity values obtained by replicate
measurements on the same or similar objects under specified conditions
Note 1 to entry: Measurement precision is usually expressed numerically by measures of imprecision, such as
standard deviation, variance, or coefficient of variation under the specified conditions of measurement.
5

---------------------- Page: 7 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

Note 2 to entry: The ‘specified conditions’ can be, for example, repeatability conditions of measurement,
intermediate precision conditions of measurement, or reproducibility conditions of measurement (see
ISO 5725-1) [2].
Note 3 to entry: Measurement precision is used to define measurement repeatability, intermediate measurement
precision, and measurement reproducibility.
Note 4 to entry: Sometimes 'measurement precision' is erroneously used to mean 'measurement accuracy'.
[SOURCE: ISO IEC Guide 99:2007, 2.15]
3.7
repeatability
precision under repeatability conditions
Note 1 to entry: Repeatability can be expressed quantitatively in terms of the dispersion characteristics of the
results.
[SOURCE: ISO 3534-2:2006, 3.3.5]
3.8
repeatability limit
r
repeatability critical difference for a specified probability of 95 %
[SOURCE: ISO 3534-2:2006, 3.3.9]
3.9
repeatability standard deviation
standard deviation of test results or measurement results obtained under repeatability conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test or measurement results under
repeatability conditions.
Note 2 to entry: Similarly, “repeatability variance” and “repeatability coefficient of variation” can be defined and
used as measures of the dispersion of test or measurement results under repeatability conditions.
[SOURCE: ISO 3534-2:2006, 3.3.7]
3.10
reproducibility
precision under reproducibility conditions
Note 1 to entry: Reproducibility can be expressed quantitatively in terms of the dispersion characteristics of the
results.
Note 2 to entry: Results are usually understood to be corrected results.
[SOURCE: ISO 3534-2:2006, 3.3.10]
6

---------------------- Page: 8 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

3.11
reproducibility limit
R
reproducibility critical difference for a specified probability of 95 %
[SOURCE: ISO 3534-2:2006, 3.3.14]
3.12
reproducibility standard deviation
standard deviation of test results or measurement results obtained under reproducibility conditions
Note 1 to entry: It is a measure of the dispersion of the distribution of test or measurement results under
reproducibility conditions.
Note 2 to entry: Similarly, “reproducibility variance” and “reproducibility coefficient of variation” can be defined
and used as measures of the dispersion of test or measurement results under reproducibility conditions.
[SOURCE: ISO 3534-2:2006, 3.3.12]
4 Principle
In order to check for the presence of PCP, a test portion of sample material is fortified with internal
standards (13C-PCP). The test portion is extracted using a QuEChERS approach for animal feed and FAD.
A ‘reversed’ QuEChERS approach is used for guar gum. For FAD samples, lipids are removed prior to
QuEChERS extraction by the addition of sulfuric acid.
Final extracts from all matrices are analysed by liquid chromatography coupled to triple quadrupole
mass spectrometry, operated in negative electrospray ionization mode.
Identification is done on the basis of retention time and mass. Quantification is done using the internal
standard method.
5 Reagents and materials
5.1 General
Use only reagents of recognized analytical grade and with a purity suitable for OC residue analysis.
Check the purity of the reagents by performing a blank test under the same conditions as used in the
method. The chromatogram should not show any interfering impurity at the retention time of
compounds of interest.
5.2 Chemicals
5.2.1 Diethylamine (DEA)
5.2.2 Acetonitrile
5.2.3 Methanol
5.2.4 Acetone
5.2.5 n-Hexane
7

---------------------- Page: 9 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

5.2.6 Sulfuric acid
5.2.7 Deionized water
5.2.8 Sodium hydroxide
5.2.8.1 Sodium hydroxide solution (10 M)
Weigh 4 g of sodium hydroxide (5.2.8) and add 10 ml deionized water (5.2.7) to achieve a concentration
of 10 M. Store at room temperature in a closed glass bottle.
5.2.9 Magnesium sulphate
5.2.10 Sodium chloride
5.2.11 Formic acid
5.2.12 Ammonium formate
5.2.12.1 Ammonium formate solution (1 M)
Weigh 6,3 g ammonium formate (5.2.12) and add 100 ml deionized water (5.2.7) to achieve a
concentration of 1 M. Store the solution at room temperature. The solution is tenable under these
conditions during at least 1 month.
5.2.13 Mobile phase A
Take 1 ml ammonium formate solution (1 M) (5.2.12.1) and add 999 ml deionized water (5.2.7) and
20 µL formic acid (5.2.11). Mix well. Store the solution at room temperature. The solution is tenable
under these conditions during at least 1 month.
5.2.14 Mobile phase B
Take 1 ml ammonium formate solution (1 M) (5.2.12.1) and add 50 ml deionized water (5.2.7), 949 ml
methanol (5.2.3) and 20 µl formic acid (5.2.11) and mix well. Store the solution at room temperature.
The solution is tenable under these conditions during at least 1 month.
5.2.15 Pentachlorophenol (PCP sodium salt, 95% purity)
5.2.15.1 PCP Stock solution 1 (2000 µg/ml)
Weigh 25 mg (± 0,01 mg) of PCP (5.2.15) (taking impurities into consideration) and add 12,5 ml
deionized water (5.2.7) to achieve a concentration of 2000 µg/ml. Store the solution in a refrigerator at
4 °C (± 3 °C). The solution is tenable under these conditions during at least 12 months.
5.2.15.2 PCP Stock solution 2 (1 ng/µl)
Dilute 50 µl of PCP Stock solution 1 (5.2.15.1) to 100 ml deionized water (5.2.7) in a volumetric flask
(5.2.20). Store the solution in a refrigerator at 4 °C (± 3 °C). The solution is tenable under these
conditions during at least 12 months.
5.2.15.3 PCP Working solution 1 (0,1 ng/µl)
Take 100 µL of PCP Stock solution 2 (5.2.15.2) and add 900 µl acetonitrile (5.2.2). Prepare this solution
daily.
8

---------------------- Page: 10 ----------------------
oSIST prEN 17362:2019
prEN 17362:2019 (E)

5.2.15.4 PCP Working solution 2 (0,01 ng/µl)
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.