CEN/TC 454 - Algae and algae products

This document specifies a laboratory method for the determination of chlorophyll a content in algae. The method was initially tested and evaluated on the microalgae species Nannochloropsis sp. and a heat treated algal product tomato soup with Nannochloropsis sp. supplement, and the macro algae species Ulva sp., Furcellaria lumbricalis, and Saccharina latissima. During an Interlaboratory Trial the method was tested on the microalgae species Nannochloropsis sp. and the macro algae species Saccharina latissima. The microalgae species Nannochloropsis sp. and Phaodactlylum sp. and the macro algae species Ulva sp. and Saccharina latissima were tested in a Round Robin test. This document is only validated for chlorophyll a, but it can be used for other chlorophylls as well.

This document defines the terms related to functions, products, and properties of algae and algae products. In order to better pack the methodologies, algae are regarded as a functional group of organisms consisting of microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes.

This document specifies methods for the measurement of energy content and main elements balances of algae from cultivation or from wild growth and algae products to provide biomass, intended for renewable algal raw material used as bioenergy and in bio-based products.
This document also specifies carbon source parameters specific to algae as bio-based and it is applicable to studies covering algae production life cycle assessment (LCA) e.g. algal biomass farming or wild collection.
This document does not apply to methods of algae and algae products sampling, harvesting and pre/postprocessing.
This document does not apply to algae and algae products intended for the food and feed sector.

This document specifies a set of principles and rules that algae producers, algae products industries, laboratories or other entities that collect algae and algae products samples can follow for the definition of their own sampling programs and sampling protocols.
In the context of this document, algae are a functional group that include microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes.
As algae and their production processes are so diverse, this document does not define a specific sampling program and/or a specific sampling protocol. Instead, this document specifies the aspects that can be considered when defining its own sampling program and protocol.
This document describes when, where and how to draw a representative sample. For guidance on sample preparation of dry and wet samples of micro- and macroalgae, and algae products, please refer to EN 17605.
This document is to be used for the collection of samples for lot characterization for commercial or legal/regulatory purposes. However, this document can also be used for any type of sampling of algae, including samples for quality control during production.

This document specifies a laboratory method for the determination of the total lipid content in micro- and macroalgae by the Ryckebosch-Foubert method.

This document specifies the sample preparation of dry and wet samples of algae and algae products. This document enables laboratories analysing algae samples to report accurate dry weight percentages and to obtain representative samples possible for further examination.

This document describes product specifications, product characteristics and other relevant information for algae and algae products for food, nutraceutical and animal feed applications. This document is a general overview of available limits, procedures and analytical methods applicable to algae and algae products used for food and feed applications.
This document does not apply to pharmaceutical, cosmetics, fertilizer/biostimulants, chemical and biofuel applications.

This document specifies a method for the detection and identification of microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes by using morphological methods and/or molecular methods.
The morphological methods in this document are applicable to harvested wet biomass and to harvested dried unground biomass from microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes that have been grown and/or harvested for further processing and/or use.
The molecular methods in this document are applicable to harvested wet biomass and to harvested dried and/or ground biomass from microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes that have been grown and/or harvested for further processing and/or use.
This document describes a toolbox, consisting of several identification methods that can be chosen according to the applicability and purpose of the identification:
-   morphological methods based on observation and referring to scientific literature on taxonomy:
-   macroscopic identification;
-   light microscopic identification.
-   molecular methods for sequencing and blasting of sequences:
-   16S rDNA sequencing;
-   18S rDNA sequencing;
-   rbcL DNA sequencing;
-   ITS sequencing;
-   COX1 gene sequencing;
-   tufA gene sequencing.
This document does not deal with genetic purity of the biomass or quantification of the identified taxa.
This document is not suitable for the analysis of highly processed biomass with highly degraded DNA where the fragments’ length are not sufficient for amplification of the targets and the morphological characteristics cannot be assessed.

This document specifies the methods to be used for the determination of productivity of algae growth sites.
This document excludes methods for sampling, harvesting and pre-/postprocessing. Excluded as well is “wild growth”, which is defined as algae growing in nature without human interference except when harvesting the algae.

The purpose of this document is to provide an overview on how quality indicating parameters for algae and algae products and intermediates relevant for chemical and bioenergy applications can be handled and to identify the need for future standards development for chemicals, bioenergy and biofuels applications.
This document does not provide instructions on handling of technical requirements in existing legislations.

This document gives an overview of recommendations on product specifications, and other relevant information, for algae and algae products for cosmetics industry.
This document does not apply to food and feed applications.
This document does not provide instructions on handling of technical requirements in existing legislations.

This document gives an overview of recommendations on product specifications, and other relevant information, for algae and algae products for pharmaceutical applications.
This document does not apply to food and feed applications.
This document does not provide instructions on handling of technical requirements in existing legislations.

This document describes a method for the determination of inorganic arsenic in algae and algae products by anion-exchange HPLC-ICP-MS following water bath extraction. The method is specifically designed for seaweeds containing inorganic arsenic and arsenosugar 408 (i.e. sulfate-arsinoylriboside). The peaks of inorganic arsenic and arsenosugar 408 are separated by gradient elution.
The method was initially tested and evaluated on the algae species Ascophyllum nodosum, Fucus vesiculosus and Saccharina latissima. Given the limited number of participating laboratories in the interlaboratory studies, this document is only validated for Ascophyllum nodosum and Saccharina latissima, but it can also be used for other algae species.

This document describes the application of Kjeldahl and Dumas methods for the determination of nitrogen content in algae and their relevant products.
The method was initially tested and evaluated on the algae species Nannochloropsis sp. and Palmaria palmata. The study validated this document for both algae species. This method can also be used for other algae species.

This document specifies a method for the quantitative determination of total uronic acids by High Performance Anion Exchange Chromatography coupled with Pulsed Amperometric Detection (HPAEC-PAD) after acid hydrolysis of the samples for algae and algae products. It specifies a method for the determination in one single analysis of mannuronic, glucuronic and guluronic acids in brown seaweed, and mannuronic and guluronic acids in alginate products. The sum of the individual uronic acid values is used for determining the total uronic acid content.

This 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 enables 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 will be available in relative (in %) and, by means of an appropriate internal standard, absolute (mg/g dw) terms. Moreover, the method described in this standard ensures a practical and safe technical approach, whose protocol details and all related know-how will be easily and economically transferrable to all the sector stakeholders. This document ensures 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 avoids 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.

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.

This document specifies a laboratory method for the quantification of phycocyanin content in the genus Arthrospira (Spirulina) by a
spectrophotometric method.

This European standard specifies a laboratory method for the determination of the
carotenoid  content in microalgae. The method is based on the development of the
chlorophyll a standard, which resulted in prEN 18034. This method has been validated
for the microalgae species  Nannochloropsis and Phaodactlylum. This standard is only
validated for fucoxanthin, beta carotene and lutein content, but could be used for other
carotenoids as well. Given small adaptations to be able to measure smaller
concentrations, this method could also be used for macro algae.
This standard is developed in CEN/TC 454/WG 6 Product test methods.

This Technical Report describes quality designations and indications for algae and directly derived products from algae production required for or by food/feed/nutraceuticals/animal food producers and industry. This TR does not apply to pharmacuetical, cosmetics and chemical applications.  
Note: This TR does not provide instructions on existing handling of technical requirements in existing legislations.

This document defines the terms related to functions, products, and properties of algae and algae products. In order to better pack the methodologies, algae are regarded as a functional group of organisms consisting of microalgae, macroalgae, cyanobacteria and Labyrinthulomycetes.