SIST EN 17899:2024

Water quality - Spectrophotometric determination of chlorophyll-a content by ethanol extraction for the routine monitoring of water quality

This document specifies a spectrophotometric method for determining the chlorophyll-a content as a measure of the amount of phytoplankton in surface water. Assuming a maximum sample volume of 2 l, chlorophyll-a content of 5 μg/l or more can be determined.

Wasserbeschaffenheit - Spektrophotometrische Bestimmung des Chlorophyll-a-Gehalts durch Ethanolextraktion für das Routinemonitoring der Wasserqualität

Dieses Dokument legt ein spektrophotometrisches Verfahren zur Bestimmung des Chlorophyll a-Gehalts als Maß für die Menge an Phytoplankton für alle Arten von Wasser fest. Bei einem angenommenen größten Probenvolumen von 2 l können Chlorophyll a-Gehalte von 5 µg/l oder mehr bestimmt werden. Die Bestimmungsgrenze kann von jedem Labor individuell berechnet werden und kann bis zu 0,5 µg/l bei Verwendung einer Probe von 2 l (oder mehr) und einer 50 mm-Küvette betragen.

Qualité de l'eau - Détermination spectrophotométrique de la teneur en chlorophylle a par extraction à l'éthanol pour la surveillance de routine de la qualité de l'eau

Le présent document décrit une méthode spectrophotométrique pour déterminer la teneur en chlorophylle a comme mesure de la quantité de phytoplancton dans tous les types d’eaux. En supposant que le volume d'échantillon maximal est de 2 l, des valeurs de teneur en chlorophylle a de 5 µg/l ou plus peuvent être déterminées. La limite de détermination peut être calculée par chaque laboratoire individuellement et peut atteindre 0,5 µg/l en utilisant 2 l d’échantillon (voire plus) et une cuve de 50 mm.

Kakovost vode - Spektrofotometrijsko določevanje klorofila-a po ekstrakciji z etanolom za rutinski monitoring kakovosti vode

General Information

• Status: Published
• Public Enquiry End Date: 02-Nov-2022
• Publication Date: 11-Nov-2024
• ICS: 13.060.70 - Examination of biological properties of water
• ICS: 71.040.50 - Physicochemical methods of analysis
• Technical Committee: KAV - Water quality
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 27-Sep-2024
• Due Date: 02-Dec-2024
• Completion Date: 12-Nov-2024
• Ref Project: EN 17899:2024 - Water quality - Spectrophotometric determination of chlorophyll-a content by ethanol extraction for the routine monitoring of water quality

Standards Content (Sample)

SLOVENSKI STANDARD
01-december-2024
Kakovost vode - Spektrofotometrijsko določevanje klorofila-a po ekstrakciji z
etanolom za rutinski monitoring kakovosti vode
Water quality - Spectrophotometric determination of chlorophyll-a content by ethanol
extraction for the routine monitoring of water quality
Wasserbeschaffenheit - Spektrophotometrische Bestimmung des Chlorophyll-a-Gehalts
durch Ethanolextraktion für das Routinemonitoring der Wasserqualität
Qualité de l'eau - Détermination spectrophotométrique de la teneur en chlorophylle a par
extraction à l'éthanol pour la surveillance de routine de la qualité de l'eau
Ta slovenski standard je istoveten z: EN 17899:2024
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN 17899
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2024
EUROPÄISCHE NORM
ICS 13.060.70; 71.040.50
English Version
Water quality - Spectrophotometric determination of
chlorophyll-a content by ethanol extraction for the routine
monitoring of water quality
Qualité de l'eau - Détermination spectrophotométrique Wasserbeschaffenheit - Bestimmung des Chlorophyll-
de la teneur en chlorophylle a par extraction à l'éthanol a-Gehalts durch Ethanolextraktion für die
pour la surveillance de routine de la qualité de l'eau routinemäßige Überwachung der Wasserqualität
This European Standard was approved by CEN on 10 June 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 17899:2024 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 . 7
5 Interferences . 7
6 Reagents . 7
7 Apparatus . 8
8 Procedure. 9
8.1 General . 9
8.2 Sample pre-treatment . 9
8.3 Filtration . 9
8.4 Extraction . 9
8.5 Measurement . 10
9 Quality assurance . 10
9.1 Blank values . 10
9.2 pH after acidification . 10
9.3 Wavelength setting . 10
9.4 Ratio A/A' . 11
10 Calculations . 11
11 Expression of results . 12
12 Test report . 12
Annex A (informative) Performance data . 13
A.1 Interlaboratory trial study in the Netherlands: . 13
A.2 Validation interlaboratory trial in Germany: . 13
Annex B (informative) Theoretical backgrounds . 16
B.1 Meaning of the determination of the chlorophyll-a content . 16
B.2 Acidifying extracts . 16
B.3 Calculation of Phaeopigment concentration . 16
Annex C (informative) Control samples . 17
C.1 Introduction . 17
C.2 Surface water . 17
C.3 Frozen spinach extracts . 17
C.4 Freeze-dried kale . 17
C.5 Algae culture. 18
Annex D (informative) SCOR-Unesco method for determination of chlorophyll in sea-water . 19
D.1 Measurement of chlorophyll a, b, c and d with SCOR-Unesco method using chromatic
equations . 19
D.2 Performance characteristics and comparison of methods . 20
Bibliography . 21
European foreword
This document (EN 17899:2024) has been prepared by the Technical Committee CEN/TC 230 “Water
analysis”, the secretariat of which is held by DIN.
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 January 2025, and conflicting national standards shall
be withdrawn at the latest by January 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.
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
Chlorophyll-a is the most common essential photosynthetic pigment present in photoautotrophic
plankton organisms. It is the main component of the dynamically regulated photosystem of these
organisms, in which other accessory pigments are involved, some of which are chemically very similar to
chlorophyll-a.
The chlorophyll-a content depends on the species composition of the phytoplankton, the time of day and
season, the place and the depth of sampling. It is also suitable for quantifying the change in the algal
biomass (cell proliferation) in biological tests to check the toxicity of substances dissolved in water.
The chlorophyll concentration of a water sample can provide information about the trophic state of a
water body. It is used as an easily determinable measure of the phytoplankton biomass and is a key
variable in many trophy scoring systems. Even if this value cannot be used as an absolute measure for the
phytoplankton biomass, the determination of the chlorophyll-a concentration together with other
biomass and bioactivity parameters provides information about the quantitative occurrence and the
potential metabolic performance of the phytoplankton in waters.
Due to the sensitivity of chlorophyll to light, acids and enzymes, there is currently no universally
applicable routine analytical method that enables an accurate, artefact-free and at the same time simple
determination of the chlorophyll-a content in water samples containing phytoplankton. The extractive,
spectrophotometric method described in this document therefore provides an operationally defined
value. As extracting agent hot ethanol is used. Various other extractants (e.g. acetone or methanol) are
described in the literature, but these may have lower extraction efficiency or are toxicologically
problematic.
WARNING — Persons using this document should be familiar with usual laboratory practice. This
document does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document be
carried out by suitably qualified staff.
Annexes A, B, C and D of this document are for information only.
1 Scope
This document describes a spectrophotometric method for determining the chlorophyll-a content
corrected for phaeopigments as a measure of the amount of phytoplankton for all types of surface water
including marine water. The determination limit is usually 2 µg/l to 5 µg/l and is calculated by each
laboratory individually. It can be as low as 0,5 µg/l using 2 l of sample (or even more) and a 50 mm
cuvette.
NOTE In some measurement programs like marine studies on time series data and ecological
status/classification no correction for phaeopigments is used and acidification is omitted, e.g. as recommended by
OSPAR.
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 5667-3, Water quality — Sampling — Part 3: Preservation and handling of water samples
(ISO 5667-3)
3 Terms and definitions
For the purpose
...