EN 17294:2019
(Main)Animal feeding stuffs - Methods of sampling and analysis - Determination of organic acids by Ion Chromatography with Conductivity Detection (IC-CD)
Animal feeding stuffs - Methods of sampling and analysis - Determination of organic acids by Ion Chromatography with Conductivity Detection (IC-CD)
This document specifies a method for the determination of organic acids in animal feeding stuffs by Ion Chromatography with conductivity detection (IC-CD).
The method is intended to be used for the determination of formic acid, lactic acid, propionic acid, citric acid, fumaric acid and malic acid as active substances in feed additives, premixtures, feed materials, compound feed and water and for acetic acid in a limited manner in the same matrices. This method determines the total extractable concentration of the above mentioned organic acids and their salts.
It is advisable that the user of this standard determines the working range of the method for each organic acid. The lower limit of the working range depends on the matrix and the interferences encountered. It is advisable that a working range between 10 mg/l and 100 mg/l is achievable.
The method was successfully tested in an inter-laboratory study in concentrations between 0,02 % up to 27 % of the above mentioned organic acids.
NOTE Limitation occurs during simultaneous determination of high concentration of lactic acid and low concentration of acetic acid. If the ratio of concentration of lactic acid to acetic acid exceeds factor 20, the determination of acetic acid is not guaranteed.
On the basis of the referred working range, sample weight and extraction volume, limits of quantification (LOQ), as calculated (Table 1) can be achievable.
Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung organischer Säuren mittels Ionenchromatographie mit Leitfähigkeitsdetektion (IC-CD)
Dieses Dokument legt ein Verfahren zur Bestimmung organischer Säuren in Futtermitteln mittels Ionen-chromatographie mit Leitfähigkeitsdetektion (en: IC-CD, ion chromatography with conductivity detection) fest.
Das Verfahren ist für die Verwendung zur Bestimmung von Ameisensäure, Milchsäure, Propionsäure, Zitronensäure, Fumarsäure und Apfelsäure als Wirkstoffe in Futtermittelzusatzstoffen, Vormischungen, Futtermittel-Ausgangserzeugnissen, Mischfuttermitteln und Wasser sowie für Essigsäure in begrenztem Maße in denselben Matrices vorgesehen. Dieses Verfahren ermittelt die extrahierbare Gesamtkonzentration der vorstehend aufgeführten organischen Säuren und ihrer Salze.
Dem Anwender dieser Norm wird empfohlen, den Arbeitsbereich des Verfahrens für jede organische Säure zu ermitteln. Der untere Grenzwert des Arbeitsbereiches hängt von der Matrix und den aufgetretenen Störungen ab. Es wird empfohlen, dass ein Arbeitsbereich zwischen 10 mg/l und 100 mg/l eingehalten wird.
Das Verfahren wurde im Rahmen einer Ringversuchsstudie bei Konzentrationen von 0,02 % bis zu 27 % der vorgenannten organischen Säuren erfolgreich geprüft.
ANMERKUNG Bei der gleichzeitigen Bestimmung einer hohen Konzentration von Milchsäure und einer niedrigen Konzentration von Essigsäure tritt eine Einschränkung auf. Wenn das Verhältnis der Konzentration der Milchsäure zu der von Essigsäure den Faktor 20 überschreitet, ist die Bestimmung der Essigsäure nicht sichergestellt.
Auf der Grundlage des zugehörigen Arbeitsbereichs, der Probenmasse und des Extraktionsvolumens können Bestimmungsgrenzen (en: LOQ, limits of quantification) wie berechnet (Tabelle 1) erreichbar sein.
Aliments des animaux - Méthodes d’échantillonnage et d’analyse - Dosage des acides organiques par chromatographie ionique avec détection conductimétrique (CI-DC)
Le présent document spécifie une méthode de dosage des acides organiques dans les aliments pour animaux par chromatographie ionique avec détection conductimétrique (CI-DC).
La méthode est destinée à permettre le dosage de l’acide formique, de l’acide lactique, de l’acide propionique, de l’acide citrique, de l’acide fumarique et de l’acide malique présents comme substances actives dans les additifs pour l’alimentation animale, les prémélanges, les matières premières pour aliments des animaux, les aliments composés pour animaux, ainsi que l’eau et l’acide acétique de manière limitée dans les mêmes matrices. Cette méthode détermine la concentration totale extractible des acides organiques susmentionnés et de leurs sels.
Il est recommandé à l’utilisateur de la présente norme de déterminer le domaine de mesure de la méthode pour chaque acide organique. La limite inférieure du domaine de mesure dépend de la matrice et des interférences rencontrées. Il est recommandé de pouvoir atteindre un domaine de mesure compris entre 10 mg/l et 100 mg/l.
La méthode a été soumise à essai avec succès lors d’une étude interlaboratoires à des concentrations allant de 0,02 % à 27 % pour les acides organiques susmentionnés.
NOTE La détermination simultanée d’une concentration élevée d’acide lactique et d’une faible concentration d’acide acétique présente des limites. Si le rapport de la concentration d’acide lactique à la concentration d’acide acétique dépasse un facteur de 20, le dosage de l’acide acétique n’est pas garanti.
En s’appuyant sur le domaine de mesure mentionné, le poids de l’échantillon et le volume d’extraction, il est possible d’atteindre les limites de quantification (LQ), telles que calculées (Tableau 1).
Krma - Metode vzorčenja in analize - Določevanje organskih kislin z ionsko kromatografijo in detekcijo na osnovi prevodnosti (IC-CD)
Ta dokument določa metodo za določevanje organskih kislin v krmi z ionsko kromatografijo in detekcijo na osnovi prevodnosti (IC-CD).
Ta metoda je predvidena za določevanje mravljične kisline, mlečne kisline, propionske kisline, citronske kisline, fumarne kisline in jabolčne kisline kot aktivnih snovi v dodatkih za krmo, premiksih, krmnih mešanicah in vodi ter za varnostne preglede ocetne kisline pri istih matrikah. Ta metoda določa skupno koncentracijo omenjenih organskih kislin in njihovih soli, ki jo je mogoče ekstrahirati.
Uporabnik tega standarda mora določiti delovni razpon metode za vsako organsko kislino. Spodnja meja delovnega razpona je odvisna od matrice in motenj, ki se pojavijo. Delovni razpon med 10 [mg/l] in 100 [mg/l] naj bi bil dosegljiv.
Metoda je bila uspešno preskušena v medlaboratorijski študiji pri koncentracijah od 0,02 % do 27 % omenjenih organskih kislin.
OPOMBA: Omejitev se pojavi pri hkratnem določevanju velike koncentracije mlečne kisline in majhne koncentracije ocetne kisline. Če razmerje med koncentracijo mlečne in ocetne kisline presega faktor 20, potem določevanje ocetne kisline ni zagotovljeno.
Na podlagi omenjenega delovnega razpona, teže vzorca in volumna ekstrakta naj bi bile mejne vrednosti kvantifikacije (LOQ) dosegljive, kot so izračunane (preglednica 1).
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-oktober-2019
Krma - Metode vzorčenja in analize - Določevanje organskih kislin z ionsko
kromatografijo in detekcijo na osnovi prevodnosti (IC-CD)
Animal feeding stuffs - Methods of sampling and analysis - Determination of organic
acids by Ion Chromatography with Conductivity Detection (IC-CD)
Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung organischer
Säuren mittels Ionenchromatographie mit Leitfähigkeitsdetektion (IC-CD)
Aliments des animaux - Méthodes d’échantillonnage et d’analyse - Dosage des acides
organiques par chromatographie ionique avec détection conductimétrique (CI-DC)
Ta slovenski standard je istoveten z: EN 17294:2019
ICS:
65.120 Krmila Animal feeding stuffs
71.040.40 Kemijska analiza Chemical analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17294
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2019
EUROPÄISCHE NORM
ICS 65.120; 71.040.40
English Version
Animal feeding stuffs - Methods of sampling and analysis -
Determination of organic acids by Ion Chromatography
with Conductivity Detection (IC-CD)
Aliments des animaux - Méthodes d'échantillonnage et Futtermittel - Probenahme- und
d'analyse - Dosage des acides organiques par Untersuchungsverfahren - Bestimmung organischer
chromatographie ionique avec détection Säuren mittels Ionenchromatographie mit
conductimétrique (CI-DC) Leitfähigkeitsdetektion (IC-CD)
This European Standard was approved by CEN on 24 June 2019.
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
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17294:2019 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 6
4 Principle . 6
5 Reagents . 6
6 Apparatus . 8
7 Sampling . 11
8 Preparation of test sample . 11
9 Procedure. 11
9.1 General . 11
9.2 Calibration . 13
9.3 Measurement of the calibration standards and sample solutions . 13
9.4 Confirmation of identity . 13
10 Calculation and expression of results . 14
11 Precision . 14
11.1 Interlaboratory test . 14
11.2 Repeatability . 15
11.3 Reproducibility . 15
12 Test report . 19
Annex A (normative) Flow Chart . 20
Annex B (informative) Examples of measuring equipment and appropriate eluents . 21
Annex C (informative) Precision data . 25
Bibliography . 33
European foreword
This document (EN 17294:2019) 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 European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by February 2020, and conflicting national standards shall
be withdrawn at the latest by Feburary 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 European Standard: 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.
Introduction
Organic acids and their salts such as citric acid, formic acid, lactic acid, acetic acid, propionic acid, fumaric
acid, benzoic acid and sorbic acid are animal feed additives which play an important role in animal feeding
by improving the animals’ performance and decreasing the development of (pathogenic) microorganisms
in the intestine especially in the pig production. Concerning the feed legislation the substances can be
used for different purposes depending on its functions and properties. According to their functional
principle or their function, the relevant organic acid could be allocated within one or more of the
functional groups mentioned in Annex I of Regulation (EC) no. 1831/2003 as preservative, acidity
regulators, flavouring compounds, silage additives or other zoo-technical additives.
1 Scope
This document specifies a method for the determination of organic acids in animal feeding stuffs by Ion
Chromatography with conductivity detection (IC-CD).
The method is intended to be used for the determination of formic acid, lactic acid, propionic acid, citric
acid, fumaric acid and malic acid as active substances in feed additives, premixtures, feed materials,
compound feed and water and for acetic acid in a limited manner in the same matrices. This method
determines the total extractable concentration of the above mentioned organic acids and their salts.
It is advisable that the user of this standard determines the working range of the method for each organic
acid. The lower limit of the working range depends on the matrix and the interferences encountered. It is
advisable that a working range between 10 mg/l and 100 mg/l is achievable.
The method was successfully tested in an inter-laboratory study in concentrations between 0,02 % up to
27 % of the above mentioned organic acids.
NOTE Limitation occurs during simultaneous determination of high concentration of lactic acid and low
concentration of acetic acid. If the ratio of concentration of lactic acid to acetic acid exceeds factor 20, the
determination of acetic acid is not guaranteed.
On the basis of the referred working range, sample weight and extraction volume, limits of
quantification (LOQ), as calculated (Table 1) can be achievable.
Table 1 — Limits of quantification (LOQ)
Organic acid LOQ
mg/kg
Formic acid 200
Lactic acid 200
Propionic acid 200
Citric acid 200
Fumaric acid 200
Acetic acid 200
Malic acid 200
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 3696, Water for analytical laboratory use — Specification and test methods (ISO 3696)
EN ISO 6498, Animal feeding stuffs — Guidelines for sample preparation (ISO 6498)
EN ISO 10304-1, Water quality — Determination of dissolved anions by liquid chromatography of ions —
Part 1: Determination of bromide, chloride, fluoride, nitrate, nitrite, phosphate and sulfate (ISO 10304-1)
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
feed additives
substances, micro-organisms or preparations, other than feed material and premixtures, which are
intentionally added to feed or water
[SOURCE: Regulation (EC) No 1831/2003/Article 2/2 a [1]]
3.2
animal feeding stuffs
any substance or product,including additives, whether processed, partially processed or unprocessed,
intended to be used for oral feeding to animals
[SOURCE: Regulation (EC) No 178/2002/Article 2/4 [2]]
4 Principle
The sample is extracted with water. The extract is filtered or centrifuged and – if necessary – diluted. The
amount of organic acids extracted from the sample is determined with ion chromatography (IC) in
conjunction with conductivity detection (CD) using external calibration.
When using CD it is essential that the eluent shows a sufficiently low conductivity. For this reason, CD is
usually combined with a suppressor device (cation exchanger), which will reduce the conductivity of the
eluent and transform the sample acids into their respective salts.
The method description follows a proven approach where the chromatographic resolution R shall be
checked to ensure that it complies with the required separation conditions in accordance with
EN ISO 10304-1.
WARNING — The use of this European Standard can involve hazardous materials, operations and
equipment. This standard does not purport to address all the safety problems associated with its use. It
is the responsibility of the user of this European Standard to establish appropriate safety and health
practices and determine the applicability of regulatory limitations prior to use.
5 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified.
5.1 Water, grade 1 in accordance with EN ISO 3696
5.2 Formic acids, lactic acid, propionic acid, citric acid, acetic acid, malic acid standard solution,
c = 1 000 [mg/l]
Single acids standard solutions with adequate and required specification are commercially available
(ready-to-use solutions).
5.3 Alternative preparation of stock solution based single standard substances
NOTE Differentiation between the enantiomers (D, L, DL) is not of interest.
5.3.1 Citric acid, minimum 99 % purity.
5.3.2 Malic acid, minimum 99 % purity.
5.3.3 Formic Acid, minimum 98 % purity.
5.3.4 Lactic acid, minimum 85 % purity.
NOT
...
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