oSIST prEN 18197:2025
(Main)Algae and algae products - Determination of the amino acid profile of micro- and macroalgae
Algae and algae products - Determination of the amino acid profile of micro- and macroalgae
This document describes a method for determining the amino acid profile of algal biomass.
It specifies a method for the determination, in one single analysis, of the following amino acids: alanine, arginine, aspartic acid (combined with asparagine), cystine (dimer of cysteine, combined with cysteine), glutamic acid (combined with glutamine), glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine and valine.
This method does not apply to the determination of tryptophan. The existing draft standard ISO/DIS 4214 – Milk and milk products – Determination of amino acids in infant formula and other dairy products will be evaluated and adapted.
Algen und Algenprodukte - Bestimmung des Aminosäuren Profils von Mikro- und Makroalgen
Dieses Dokument beschreibt ein Verfahren zur Bestimmung des Aminosäurenprofils von Mikro- und Makroalgen.
Es legt ein Verfahren für die quantitative Bestimmung der folgenden Aminosäuren in einer einzigen Analyse fest: Alanin, Arginin, Asparaginsäure (kombiniert mit Asparagin), Cystin (Dimer von Cystein, kombiniert mit Cystein), Glutaminsäure (kombiniert mit Glutamin), Glycin, Histidin, Isoleucin, Leucin, Lysin, Methionin, Phenylalanin, Prolin, Serin, Threonin, Tyrosin und Valin.
Dieses Verfahren gilt nicht für die Bestimmung von Tryptophan.
Algues et produits à base d'algues - Détermination du profil en acides aminés des micro et macroalgues
Le présent document décrit une méthode de détermination du profil en acides aminés des micro- et macroalgues.
Il spécifie une méthode pour la détermination quantitative, en une seule analyse, des acides aminés suivants : alanine, arginine, acide aspartique (combiné à l’asparagine), cystine (dimère de la cystéine, combiné à la cystéine), acide glutamique (combiné à la glutamine), glycine, histidine, isoleucine, leucine, lysine, méthionine, phénylalanine, proline, sérine, thréonine, tyrosine et valine.
Cette méthode ne s’applique pas à la détermination du tryptophane
Alge in izdelki iz alg - Določanje aminokislinskega profila mikro- in makroalg
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-julij-2025
Alge in izdelki iz alg - Določanje aminokislinskega profila mikro- in makroalg
Algae and algae products - Determination of the amino acid profile of micro- and
macroalgae
Algen und Algenprodukte - Bestimmung des Aminosäuren Profils von Mikro- und
Makroalgen
Algues et produits à base d'algues - Détermination du profil en acides aminés des micro
et macroalgues
Ta slovenski standard je istoveten z: prEN 18197
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
June 2025
ICS
English Version
Algae and algae products - Determination of the amino
acid profile of micro- and macroalgae
Algues et produits à base d'algues - Détermination du Algen und Algenprodukte - Bestimmung des
profil en acides aminés des micro et macroalgues Aminosäuren Profils von Mikro- und Makroalgen
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 18197:2025 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 6
5 Reagents . 7
6 Reagents and standard preparation . 8
6.1 Reagents common to UV and MS detection . 8
6.2 Reagents specific for UV detection . 10
6.3 Reagents specific for MS detection . 11
7 Apparatus . 12
8 Procedure. 13
8.1 Procedure with UV detection . 13
8.1.1 General . 13
8.1.2 Sample preparation . 13
8.1.3 Cystine Calibration standards preparation . 13
8.1.4 Hydrolysis (of samples and cystine standards) . 14
8.1.5 Neutralization and dilution (of samples and converted cystine standards) . 14
8.1.6 Amino acids calibration standards preparation (not needed to be acid hydrolysed) . 14
8.1.7 Derivatization (of samples, cystine standards and amino acids standards) . 14
8.1.8 UHPLC separation and UV detection . 15
8.1.9 Peak identification and integration . 16
8.2 Procedure with MS detection . 16
8.2.1 General . 16
8.2.2 Sample preparation . 16
8.2.3 Cystine Calibration standards preparation . 17
8.2.4 Hydrolysis (of samples and cystine standards) . 17
8.2.5 Neutralization and dilution (of samples and converted cystine standards) . 17
8.2.6 Amino acids calibration standards preparation (not needed to be acid hydrolysed) . 18
8.2.7 Derivatization (of samples, cystine standards and amino acids standards) . 18
8.2.8 UHPLC separation and MS detection . 18
8.2.9 Peak identification and integration . 21
9 Calculation and expression of results . 21
9.1 Calibration curve . 21
9.2 Amino acid calculation . 22
10 Precision . 23
10.1 General . 23
10.2 Repeatability . 23
10.3 Reproducibility . 23
11 Test report . 25
Annex A (informative) Precision data . 26
Annex B (informative) Chromatogram examples . 33
Annex C (informative) Equivalence between UV and MS methods . 35
Annex D (informative) Calculation of nitrogen-to-protein conversion factor. 38
Bibliography . 40
European foreword
This document (prEN 18197:2025) 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
This document has been prepared by the experts of CEN/TC 454 'Algae and algae products'.
The European Committee for Standardization (CEN) was requested by the European Commission (EC) 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 interest in algae and algae-based products or intermediates has increased significantly in Europe as
a valuable source including but not limited to, carbohydrates, 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 textile, cosmetics, biopolymers, biofuel and fertilizer/biostimulants. Standardization was
identified as having an important role in order to promote the use of algae and algae products.
The work of CEN/TC 454 should improve 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 will promote and support
commercialisation of the European algae industry.
Determining amino acids is important for the industry to measure specific amino acids and establish the
whole amino acid profile of algae and algae products. Moreover, total amino acids can be used to calculate
the protein content of a sample with accuracy.
The goal of this document is to give algae producers and algae products industries a recommendation for
an analysis method for amino acids which can be utilized for algae. The document relies strongly on
standards ISO 4214 | IDF 254 Milk and milk products - Determination of amino acids in infant formula and
other dairy products [1], AOAC 2018.06, Total Amino Acids in Infant Formulas and Adult Nutritionals [2],
and AACC 07-50.01, Total Amino Acids by UHPLC-UV [3], and adds the possibility to use mass
spectrometry for the determination of amino acids.
1 Scope
This document describes a method for determining the amino acid profile of micro- and macroalgae.
It specifies a method for the quantitative determination, in one single analysis, of the following amino
acids: alanine, arginine, aspartic acid (combined with asparagine), cystine (dimer of cysteine, combined
with cysteine), glutamic acid (combined with glutamine), glycine, histidine, isoleucine, leucine, lysine,
methionine, phenylalanine, proline, serine, threonine, tyrosine and valine.
This method does not apply to the determination of tryptophan.
2 Normative references
The following document is 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 17399:2024, Algae and algae products - Vocabulary
EN 17605:2022, Algae and algae products - Methods of sampling and analysis - Sample treatment
3 Terms and definitions
For the purposes of this document, the terms and definitions in EN 17399 apply.
ISO and IEC maintain terminological 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/
4 Principle
Proteins are hydrolysed in 6 mol/l hydrochloric acid (HCl) for 24 h at 110 °C in the presence of phenol
and 3-3′-dithiodipropionic acid (DDP). Phenol is added to prevent halogenation of tyrosine. DDP is added
to convert cystine and cys
...
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