prEN 18210
(Main)Algae and algae Products - Determination of the fatty acid composition
Algae and algae Products - Determination of the fatty acid composition
This document specifies a laboratory method for the determination of the absolute and relative fatty acid composition of micro and
macro algae by gas chromatography coupled to a flame ionisation detector (GC-FID) of the fatty acid methyl esters.
Algen und Algenprodukte - Bestimmung der Fettsäurezusammensetzung
Das Dokument umfasst die Bestimmung des Fettsäurenprofils von Algen und Algenprodukten, was nach den vom CEN verabschiedeten Definitionen Mikro- und Makroalgen einschließt. Diese Bestimmung muss ermöglichen, dass alle in einer maßgeblichen Menge (>1% der gesamten Fettsäuren) in der Algenmatrix enthaltenen Fettsäuren auf eine genaue und reproduzierbare Weise quantifiziert werden. Die Konzentration von jeder Fettsäure muss in relativen (in%) und, durch eine geeignete interne Standardlösung, absoluten (mg/gTrockenmasse) Werten verfügbar sein. Darüber hinaus muss das in der Norm beschriebene Verfahren einen praktischen, schnellen und sicheren technischen Ansatz sicherstellen, dessen Protokolldetails und gesamtes damit verbundenes Anwendungswissen einfach und in wirtschaftlicher Weise auf alle Stakeholder im Sektor übertragbar sein müssen. Die Norm muss dieses Ziel durch eine umfassende und ausführliche Beschreibung aller technischen Schritte von der Probe an sich (einschließlich Zustand und Form) bis zum gaschromatographischen Verfahren und zur Berechnung des Fettsäurengehalts sicherstellen. Die Formulierungen dürfen keine Risiken für Mehrdeutigkeiten oder Fehlinterpretationen bergen. Schließlich bezieht diese Verfahrensnorm Informationen aus anderen äquivalenten Normen, die auf andere Matrices angewendet werden, ein und berücksichtigt andere Normen, die die spezifische Behandlung oder das Extraktionsverfahren für die Probe vor der eigentlichen Fettsäurenanalyse betreffen.
Algues et produits à base d'algues - Détermination de la composition en acides gras
Le document couvre la détermination du profil en acides gras des algues et des produits à base d’algues, en incluant donc les micro- et macroalgues, conformément aux définitions adoptées par le CEN. Cette détermination doit permettre de quantifier de manière précise et reproductible tous les acides gras présents à un niveau significatif (> 1 % du total des acides gras) dans la matrice algale. La concentration de chaque acide gras doit être disponible en termes relatifs (en %) et, au moyen d’un étalon interne approprié, en termes absolus (mg/g de poids sec). De plus, la méthode décrite dans la norme doit garantir une approche technique pratique, rapide et sûre, dont les détails du protocole et l’ensemble du savoir-faire associé doivent être facilement et économiquement transférables à toutes les parties prenantes du secteur. La norme doit assurer cet objectif par une description complète et entièrement détaillée de toutes les étapes techniques, depuis l’échantillon lui-même (y compris son état et sa forme) jusqu’à la technique de chromatographie en phase gazeuse et le calcul de la teneur en acides gras. La formulation doit également éviter tout risque d’ambiguïté ou d’interprétation erronée. Enfin, cette norme méthodologique sera éclairée par d’autres normes équivalentes appliquées à d’autres matrices et prendra en compte d’autres normes concernant le traitement spécifique ou la procédure d’extraction de l’échantillon avant l’analyse des acides gras elle-même.
Alge in izdelki iz alg - Določanje sestave maščobnih kislin
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2025
Alge in izdelki iz alg - Določanje sestave maščobnih kislin
Algae and algae Products - Determination of the fatty acid composition
Algen und Algenprodukte - Bestimmung der Fettsäurezusammensetzung
Algues et produits à base d'algues - Détermination de la composition en acides gras
Ta slovenski standard je istoveten z: prEN 18210
ICS:
13.020.55 Biološki izdelki Biobased products
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
DRAFT
EUROPEAN STANDARD
NORME EUROPÉENNE
EUROPÄISCHE NORM
July 2025
ICS
English Version
Algae and algae Products - Determination of the fatty acid
composition
Algues et produits à base d'algues- Détermination de la Algen und Algenprodukte - Bestimmung der
composition en acides gras Fettsäurezusammensetzung
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 454.
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, 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, Türkiye 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
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 18210:2025 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Principle . 7
5 Apparatus . 7
6 Reagents and materials . 7
7 Sampling and sample handling . 8
8 Procedure . 8
8.1 General. 8
8.2 Specific procedure description . 8
8.2.1 Indirect method . 8
8.2.2 Direct method . 11
9 Calculations . 13
10 Quality control . 15
10.1 General. 15
10.2 Interlaboratory reproducibility . 15
11 Test report . 15
Annex A (informative) Results of the interlaboratory trial for fatty acids . 16
Annex B (Informative) Results of interlaboratory study for fatty acids . 48
Bibliography . 82
European foreword
This document (prEN 18210:2025) has been prepared by Technical Committee CEN/TC 454 “Algae and
algae products”, 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.
Introduction
The European Commission (EC) has requested the European Committee for Standardization (CEN) to
draft European standards or European standardization deliverables to support the implementation of
Article 3 of Directive 2009/28/EC for algae and algae-based products or intermediates. This request,
presented as Mandate M/5471, also contributes to the Communication on “Innovating for Sustainable
Growth: A Bio Economy for Europe”.
The former working group CEN Technical Board Working Group 218 “Algae” was created in 2016 to
develop a work programme as part of this Mandate. The technical committee CEN/TC 454 'Algae and
algae products' was established to carry out the work programme that will prepare a series of standards.
The importance of algae and algae-based products or intermediates has increased significantly in Europe
as these products have been shown to be a valuable source, including but not limited to, of carbohydrates
(comprising, in particular, hydrocolloids with a large commercial importance, such as agar, alginate or
carrageenan), proteins, lipids, and several pigments. These materials are suitable for use in a wide range
of applications from food and feed purposes to other sectors, such as textiles, cosmetics, cosmeceuticals,
pharmaceuticals, biopolymers, biofuel, and fertilizer/biostimulants. Standardization of analytical
methods has been highlighted as having an important role in promoting the use of algae and algae
products.
The work of CEN/TC 454 should contribute to the reliability of the supply chain, thereby improving the
confidence of industry and consumers in algae, which include macroalgae, microalgae, cyanobacteria,
Labyrinthulomycetes, algae-based products or intermediates and should promote and support
commercialization of products of the European algae industry.
In this context, the fatty acid profile of algae and algal products is an important aspect for assessing the
potential nutritional quality and biological activity of algal biomass, thus paving the way for multiple
relevant and high added-value applications.
1 Scope
The document encompasses the determination of the fatty acid profile in algae and algae products,
thereby including micro- and macroalgae, according to the definitions adopted by CEN. This
determination shall enable that all fatty acids present at a significant level (> 1 % of the total fatty acids)
in the algal matrix are quantified in an accurate and reproducible way. The concentration of each fatty
acid shall be available in relative (in %) and, by means of an appropriate internal standard, absolute
(mg/g dw) terms. Moreover, the method described in the standard shall ensure a practical, quick, and
safe technical approach, whose protocol details and all related know-how shall be easily and
economically transferrable to all the sector stakeholders. The standard shall ensure this objective by a
comprehensive and fully detailed description of all technical steps from the sample itself (including its
state and form) to the gas chromatographic technique and the calculation of the fatty acid content. The
wording shall also avoid any risk of ambiguity or wrong interpretation. Finally, this methodological
standard will be informed by other equivalent standards applied to other matrices and will take into
account other standards concerning specific treatment or extractive procedure of the sample prior to the
fatty acid analysis itself.
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 17399, Algae and algae products - Terms and definitions
EN 17605:2022, Algae and algae products - Methods of sampling and analysis - Sample treatment
EN 17908, Algae and algae products — Methods of sampling and analysis — Determination of total lipids
content using the Ryckebosch-Foubert method
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 17399 and the following apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https://www.iso.org/obp/
— IEC Electropedia: available at https://www.electropedia.org/
3,1
total lipid content
all lipid substances extracted from the test portion under the operating conditions specified, expressed
in mg/g or g/kg relative to dry weight
3,2
transesterification
chemical reaction involving the conversion of fatty acid constituents regardless of their specific chemical
form (triacylglycerols, free fatty acids, phospholipids, etc.) into fatty acid methyl esters
3,3
direct transesterification approach
methodological approach consisting of the transesterification of all fatty acid constituents present in the
sample regardless of their specific chemical form (triacylglycerols, free fatty acids, phospholipids, etc.)
3,4
indirect transesterification approach
methodological approach consisting of the transesterification of all fatty acid constituents present in the
oil extracted from the sample by the Ryckebosch method regardless of their specific chemical form
(triacylglycerols, free fatty acids, phospholipids, etc.)
3,5
derivatization
first part of the process of preparing samples to be analysed by Gas Chromatography and comprising the
steps of sample preparation for transesterification and those corresponding to the transesterification
reaction itself
3,6
fatty acid methyl ester extraction
second part of the process of preparing samples to be analysed by Gas Chromatography and comprising
the steps of extracting the fatty acid methyl esters from the transesterification reaction mixture into an
organic phase and the purification of this phase
3,7
fatty acid methyl ester analysis
final part of the analytical method corresponding to the injection of the prepared fatty acid methyl esters
in a Gas Chromatography system for detection, identification, and quantification
3,8
internal standard
fatty acid that is chemically similar to the fatty acids in algae, but it is not present in the sample, being
added in the form of methyl ester in a constant known amount to every sample prior to the derivatization
process
3,9
fatty acid methyl ester peak area
area of a specific fatty acid methyl ester in a Gas Chromatography chromatogram
3,10
relative fatty acid methyl ester peak area
share in percentage of the area of a specific fatty acid methyl ester within the sum of the areas of all fatty
acid methyl esters in a Gas Chromatography chromatogram
3,11
total relative fatty acid content
sum of all identified and significant (> 1 % of the sum of the areas of all fatty acid methyl esters) relative
fatty acid methyl ester peak areas in a Gas Chromatography chromatogram
3,12
relative fatty acid content
share in percentage of a particular relative fatty acid methyl ester peak area in the total relative fatty acid
content in the test portion under the specified operating conditions
3,13
total absolute fatty acid content
concentration of all fatty acid constituents in the algal sample corresponding to the fatty acid methyl
esters identified by Gas Chromatography and expressed in mg of fatty acid methyl ester/g of sample or g
fatty acid methyl ester/kg of sample in dry weight terms
3,14
absolute fatty acid content
concentration of a specific fatty acid constituent in the algal sample corresponding to a fatty acid methyl
ester identified by Gas Ch
...
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