EN 17706:2024
(Main)Plant biostimulants - Determination of inorganic arsenic
Plant biostimulants - Determination of inorganic arsenic
This document specifies a method for extraction, separation, and determination of inorganic arsenic (iAs) in plant biostimulants using anion-exchange high performance liquid chromatography (HPLC) or ion chromatography (IC) coupled to ICP-MS.
This document is applicable to the blends of fertilizing products where a blend is a mix of at least two of the following component EU fertilising products categories: Fertilizers, Liming Materials, Soil Improvers, Growing Media, Inhibitors, Plant Biostimulants, and where the following category Plant Biostimulants is the highest percentage in the blend by mass or volume, or in the case of liquid form by dry mass. If Plant Biostimulants is not the highest percentage in the blend, the European Standard for the highest percentage of the blend applies. In case a blend of fertilizing products is composed of components in equal quantity or in case the component EU fertilising products used for the blend have identical formulations , the user decides which standard to apply.
Pflanzen-Biostimulanzien - Bestimmung von anorganischem Arsen
Dieses Dokument legt ein Verfahren zur Extraktion, Trennung und Bestimmung von anorganischem Arsen in Pflanzen-Biostimulanzien mit Anionenaustausch-Hochleistungs-Flüssigchromatograph (HPLC) oder Ionen¬chromatographie (IC) gekoppelt mit ICP MS fest.
Dieses Dokument ist für die Mischungen von Düngeprodukten anwendbar, in denen eine Mischung aus mindestens zwei der folgenden Kategorien von Komponenten-EU Düngeprodukten besteht: Düngemittel, Kalkdünger, Bodenverbesserungsmittel, Kultursubstrate, Hemmstoffe, Pflanzen-Biostimulanzien und wenn die folgende Kategorie Pflanzen-Biostimulanzien prozentual den höchsten Massen- oder Volumenanteil in der Mischung bzw., bei flüssiger Form, den höchsten Trockenmasseanteil aufweist. Wenn Pflanzen-Biostimulanzien nicht den höchsten prozentualen Anteil in der Mischung ausmachen, ist die für den höchsten prozentualen Anteil geltende Europäische Norm anwendbar. Falls eine Düngemittelmischung aus Komponenten gleicher Menge besteht oder falls die für die Mischung verwendeten Komponenten-EU Düngeprodukte identische Formulierungen haben, entscheidet der Anwender, welche Norm angewendet wird.
Biostimulants des végétaux - Dosage de l'arsenic inorganique
Le présent document spécifie une méthode d’extraction, de séparation et de dosage de l’arsenic inorganique (iAs) dans les biostimulants des végétaux par chromatographie en phase liquide haute performance (HPLC) avec échange d’anions ou par chromatographie ionique (IC) couplée à une ICP-MS.
Le présent document est applicable aux mélanges de fertilisants constitués d’au moins deux composants appartenant aux catégories de fertilisants UE suivantes : engrais, amendements minéraux basiques, amendements du sol, supports de culture, inhibiteurs, biostimulants des végétaux et, dans lesquels la catégorie Biostimulants des végétaux représente le pourcentage le plus élevé du mélange en masse ou en volume, ou dans le cas d’un liquide, en masse sèche. Si les biostimulants des végétaux ne représentent pas le pourcentage le plus élevé du mélange, la Norme européenne relative au composant représentant le pourcentage le plus élevé du mélange s’applique. Dans le cas où un mélange de fertilisants contient des composants en quantité égale ou dans le cas où les composants des fertilisants UE utilisés pour le mélange ont des formulations identiques, l’utilisateur décide quelle norme appliquer.
Rastlinski biostimulanti - Določanje anorganskega arzena
Ta dokument določa metodo za ekstrakcijo, ločevanje in določanje anorganskega arzena (iAs) v rastlinskih biostimulantih z anionsko-izmenjevalno tekočinsko kromatografijo visoke ločljivosti (HPLC) ali ionsko kromatografijo (IC) v povezavi z masno spektrometrijo z induktivno sklopljeno plazmo (ICP-MS).
Ta dokument se uporablja za mešanice sredstev za gnojenje, ki vsebujejo najmanj dve od naslednjih kategorij sredstev za gnojenje EU: gnojila, sredstva za apnenje, izboljševalci tal, rastni substrati, inhibitorji in rastlinski biostimulanti, pri čemer rastlinski biostimulanti predstavljajo največji masni ali prostorninski odstotek mešanice ali, v primeru tekoče oblike, odstotek suhe mase. Če rastlinski biostimulanti ne predstavljajo največjega odstotka v mešanici, se uporablja evropski standard za največji odstotek mešanice. Če vsebuje mešanica sredstev za gnojenje sestavine v enakih količinah ali če imajo posamezna sredstva za gnojenje EU, uporabljena za mešanico, enake formulacije1, standard za uporabo izbere uporabnik.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-februar-2025
Nadomešča:
SIST-TS CEN/TS 17706:2023
Rastlinski biostimulanti - Določanje anorganskega arzena
Plant biostimulants - Determination of inorganic arsenic
Pflanzen-Biostimulanzien - Bestimmung von anorganischem Arsen
Biostimulants des végétaux - Dosage de l'arsenic inorganique
Ta slovenski standard je istoveten z: EN 17706:2024
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17706
EUROPEAN STANDARD
NORME EUROPÉENNE
November 2024
EUROPÄISCHE NORM
ICS 65.080 Supersedes CEN/TS 17706:2022
English Version
Plant biostimulants - Determination of inorganic arsenic
Biostimulants des végétaux - Dosage de l'arsenic Pflanzen-Biostimulanzien - Bestimmung von
inorganique anorganischem Arsen
This European Standard was approved by CEN on 26 August 2024.
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, Türkiye 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
© 2024 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17706:2024 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 . 6
5 Reagents . 6
6 Apparatus . 8
7 Sampling . 9
8 Procedure . 9
8.1 Sample preparation . 9
8.2 Water bath extraction . 9
8.3 Preparation of the calibration solutions . 9
8.4 Measurement . 9
8.4.1 General instrumental conditions . 9
8.4.2 HPLC-ICP/MS measurement of inorganic arsenic . 10
9 Calculation and expression of the results . 12
9.1 Integration of peaks . 12
9.2 Calculation of inorganic arsenic in the samples . 12
10 Test report . 13
Annex A (informative) Examples of typical chromatographic separation . 14
Annex B (informative) Results of the inter-laboratory study . 18
B.1 Inter-laboratory tests . 18
B.2 Statistical results for determination of inorganic arsenic . 18
Annex ZA (informative) Relationship of this European Standard and the essential
requirements of Regulation (EU) 2019/1009 making available on the market of EU
fertilising products aimed to be covered . 20
Bibliography . 21
European foreword
This document (EN 17706:2024) has been prepared by Technical Committee CEN/TC 455 “Plant
Biostimulants”, the secretariat of which is held by AFNOR.
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 May 2025, and conflicting national standards shall be
withdrawn at the latest by May 2025.
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 supersedes CEN TS 17706:2022.
In comparison with the previous edition no technical changes were applied in this document. The
following main editorial changes have been made:
— Introduction;
— Scope – text regarding blends changed;
— Annex B – results of the interlaboratory study added;
— Annex ZA added;
— Bibliography reordered.
This document has been prepared under a Standardization Request addressed to CEN by the European
Commission. The Standing Committee of the EFTA States subsequently approves these requests for its
Member States.
For relationship with EU Legislation, see informative Annex ZA, which is an integral part of this document.
Any feedback and questions on this document should be directed to the users’ national standards body.
A complete listing of these bodies can be found on the CEN website.
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, Türkiye and the United
Kingdom.
Introduction
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
Regulation (EU) 2019/1009 of 5 June 2019 [1] laying down rules on the making available on the market
of EU fertilising products (“FPR” or “Fertilising Products Regulation”).
This standardization request, presented as SR M/564 and relevant amendments, also contributes to the
Communication on “Innovating for Sustainable Growth: A Bio economy for Europe”. The interest in plant
biostimulants has increased significantly in Europe as a valuable tool to use in agriculture.
Standardization was identified as having an important role in order to promote the use of biostimulants.
The work of CEN/TC 455 seeks to improve the reliability of the supply chain, thereby improving the
confidence of farmers, industry, and consumers in biostimulants, and will promote and support
commercialisation of the European biostimulant industry.
The standard is based on a mild acid oxidative extraction of the arsenic species followed by liquid
chromatography (HPLC or IC) coupled to the element-specific detector ICP-MS for the determination of
the mass fraction of inorganic arsenic.
The other standards developed for determination of inorganic arsenic content in animal feedingstuffs
and foodstuffs were studied as a basis of the described method [2], [3], [4], [5].
The inter-laboratory study reflects the final statistical characteristics of the method for the determination
of inorganic arsenic content in plant biostimulants. The results are given in Annex B (informative).
WARNING — Persons using this document should be familiar with usual laboratory practice. This
document does not purport to address all of the safety issues, if any, associated with its use. It is the
responsibility of the user to establish appropriate health and safety practices and to ensure compliance
with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this document are carried out
by suitably trained staff.
1 Scope
This document specifies a method for extraction, separation, and determination of inorganic arsenic (iAs)
in plant biostimulants using anion-exchange high performance liquid chromatography (HPLC) or ion
chromatography (IC) coupled to ICP-MS.
This document is applicable to the blends of fertilizing products where a blend is a mix of at least two of
the following component EU fertilising products categories: Fertilizers, Liming Materials, Soil Improvers,
Growing Media, Inhibitors, Plant Biostimulants, and where the following category Plant Biostimulants is
the highest percentage in the blend by mass or volume, or in the case of liquid form by dry mass. If Plant
Biostimulants is not the highest percentage in the blend, the European Standard for the highest
percentage of the blend applies. In case a blend of fertilizing products is composed of components in equal
quantity or in case the component EU fertilising products used for the blend have identical formulations ,
the user decides which standard to apply.
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 17702-1:2024, Plant biostimulants — Sampling and sample preparation — Part 1: Sampling
EN 17702-2:2024, Plant biostimulants — Sampling and sample preparation — Part 2: Sample preparation
EN 17704:2024, Plant biostimulants — Determination of dry matter
3 Terms and definitions
For the purposes of this document, the following terms and definitions 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
analyte
parameter to be determined
3.2
blank calibration solution
solution prepared in the same way as the calibration solution but leaving out the analytes
3.3
blank test solution
solution prepared in the same way as the test sample solution but omitting the test portion
An example of such a blend is a product with two claimed functions consisting of a non-microbial plant
biostimulant and an organic fertilizer composed of 1kg/kg of plant biostimulant from seaweed.
3.4
calibration solution
solution used to calibrate the instrument, prepared from stock solutions by adding acids, buffer, reference
element and salts as needed
3.5
stock solution
solution with accurately known analyte concentration(s), prepared from pure chemicals
4 Principle
This document specifies a method for the determination of inorganic arsenic in plant biostimulants.
Inorganic arsenic consists of arsenite As (III) and arsenate As(V). A representative test portion of the
sample is treated with a diluted nitric acid and hydrogen peroxide solution in a heated water bath. By this
means the sample is solubilised, arsenic species are extracted into solution and As (III) is oxidized to
As(V). The inorganic arsenic is selectively separated from other arsenic compounds using anion exchange
HPLC (High Performance Liquid Chromatography) coupled online to the element-specific detector ICP-
MS (Inductively Coupled Plasma Mass Spectrometer) for the determination of the mass fraction of the
inorganic arsenic. External calibration with solvent matrix-matched standards is used for the
quantification of the amount of the inorganic arsenic. Alternatively, IC (ion chromatography) coupled to
ICP-MS can be used.
A preliminary determination of the total arsenic in aqua regia extracts by ICP-AES (EN 17701-1 [6] and
EN 17701-2 [7]) could reduce the number of the samples where the determination of iAs is necessary
because if the content of aqua regia (total) extractable arsenic is lower than the legislative limit for iAs
then the determination of iAs is not necessary.
5 Reagents
When using a method of high sensitivity like ICP-MS and HPLC the control of the blank levels of water,
acid and other reagents is very important. The reagents shall be of adequate purity and of recognized
analytical grade. The concentration of arsenic species in the reagents and water used shall be negligible
and low enough not to affect the results of the determination. Generally ultra-pure water from
a purification system and nitric acid of minimum p.a. quality is recommended.
5.1 Water with an electrical conductivity not higher than 0,1 mS/m at 25 °C, having a resistivity
greater than 18,2 MΩ⋅cm.
5.2 Nitric acid (HNO ), concentrated, ≥ 65 % (mass fraction), mass concentration of approximately
ρ(HNO ) 1,4 g/ml.
Use only nitric acid available with high purity (minimum p.a. quality) in order to avoid potential
contamination.
5.3 Hydrogen peroxide, H O not less than 30 % (mass fraction).
2 2
High purity is essential to avoid potential contamination. Commercially available hydrogen peroxide for
analysis should be tested for contamination of arsenic prior to use. It is necessary to prevent peroxide
degradation and ensure the stability of the solution, this is in the discretion of the analyst to use only
H O of adequate quality.
2 2
5.4 Extraction solution, 0,1 mol/l HNO in 3 % (volume fraction) H O .
3 2 2
Add 6,5 ml of HNO (5.2) and thereafter 100 ml of hydrogen peroxide (5.3) into 800 ml water (5.1) in
a 1 000 ml volumetric flask. Fill the flask to the mark with water (5.1.). This solution is prepared on the
day of use.
It is recommended that the total volume needed for the analysis is estimated and only this amount is
produced in the day of use.
5.5 Ammonium carbonate, (NH ) CO , mass fraction w ≥ 99,999 %, for preparation of the mobile
4 2 3
phase solution.
5.6 Aqueous ammonia, (NH (aq).) w ≥ 25 %, for adjustment of pH in the mobile phase.
5.7 Methanol, (CH OH), HPLC grade, for preparation of the mobile phase solution.
5.8 Mobile phase, e.g. 50 mmol/l ammonium carbonate in 3 % (volume fraction) methanol at pH 10,3.
Dissolve 4,80 g of ammonium carbonate (5.5) in approximately 800 ml water (5.1). Adjust the pH to 10,3
with aqueous ammonia (5.6) and add 30 ml of methanol (5.7) and then fill up to 1 000 ml with water
(5.1). Prior to use filter the mobile phase solution through a 0,45 µm filter using a filtering device (6.4).
The optimal concentration of ammonium carbonate in the mobile phase depends on the analytical column
used (e.g. brand, particle size and dimensions) and should be verified in advance. The appropriate
concentration of ammonium carbonate (usually between 10 mmol/l and 50 mmol/l) is highly dependent
on the column used and is up to the discretion of the analyst. It should fulfil the criteria for sufficient
resolution of the arsenate peak.
Methanol is added to the mobile phase in order to enhance the signal intensity for arsenic. The
concentration of methanol to achieve the highest signal to noise ratio depends on the instrument used
and should be identified by the analyst.
Different mobile phase may be also used according to the instructions of the manufacturer of the column,
but it is necessary to verify optimal separation conditions.
For example, Agilent column G3154-65001 with a guard column G3154-65002 and a mobile phase
recommended for this column, as a mixture of potassium dihydrogen phosphate (KH PO ) 2 mmol/l,
2 4
ethylenediaminetetraacetic acid disodium salt dihydrate (EDTA disodium salt) 0,2 mmol/l and the pH 6,0
adjusted with sodium hydroxide (NaOH) 1 mol/l, were successfully used for the analysis.
5.9 Arsenic (V) standard stock solution, with an arsenic (V) mass concentration of 1 000 mg/l.
The use of commercial standards of arsenic As (V), with a mass concentration of 1 000 mg/l is
recommended.
5.10 Arsenic (V) standard solution I, with an arsenic (V) mass concentration of 10 mg/l in 2 %
(volume fraction) HNO .
Pipette 1 ml of arsenic standard stock solution (5.9) into a 100 ml volumetric flask. Add 2 ml of nitric acid
(5.2), fill to the mark with water (5.1) and mix well. This solution is stable in a refrigerator at least one
week.
5.11 Arsenic (V) standard solution II, with an arsenic (V) mass concentration of 1 mg/l.
2 ®
Agilent is a registered trademark of Agilent Technologies Inc. This information is given for the convenience of
users of this document and does not constitute an endorsement by CEN of the product named.
Pipette 10 ml of arsenic standard solution I (5.10) into a 100 ml volumetric flask, fill to the mark with
water (5.1) and mix well. This solution should be prepared on the same day of use.
5.12 Solution for checking chromatographic separation, containing organic arsenic compounds (e.g.
10 μg/l) monomethylarsenous acid (MMA), dimethylarsinic acid (DMA) and arsenobetaine (AB), as well
as arsenate (e.g. 10 μg/l), arsenite (e.g. 10 μg/l) and chloride (e.g. 100 mg/l).
This solution is recommended to demonstrate satisfactory resolution of individual arsenic species,
possible interferents and to find out how the chromatographic conditions should be optimized (e.g. by
changing the mobile phase concentration or the mobile phase flow rate).
The solution shall be prepared in water (5.1), not in the extraction solution, to check the retention time
of the individual arsenic species, their visual presentation in the chromatogram and how the peaks are
separated.
6 Apparatus
6.1 Common laboratory glass (plastic) ware.
Plastic volumetric flasks are recommended for the preparation of the mobile phase and calibration
solutions. All glassware and plastic ware shall be adequately cleaned and stored to avoid any
contamination.
6.2 Laboratory grinder, capable of grinding to a particle size less than 0,5 mm.
6.3 Analytical balance, capable of weighing to an accuracy of 1 mg or better.
6.4 Filtering device, for filtration of mobile phase with a filter, pore size 0,45 µm.
6.5 Shaking heated water bath, capable of maintaining 90 °C.
Some fine materials can form a thin layer on the surface of the extraction solution and the contact of the
sample with the ext
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