Soil quality - Measurement of enzyme activity patterns in soil samples using colorimetric substrates in micro-well plates (ISO 20130:2018)

This document specifies a method for the measurement of several hydrolase activities (arylamidase, arylsulfatase, β-galactosidase, α-glucosidase, β-glucosidase, N-acetyl-glucosaminidase, acid, alkaline and global phosphatases, urease) simultaneously (or not) in soil samples, using colorimetric substrates. Enzyme activities of soil vary seasonally and depend on soil chemical, physical and biological characteristics. This method can be applied either to detect harmful effects on soil enzyme activities derived from toxic substances or other anthropogenic agents in contaminated soils against a control soil, or to test chemicals.

Bodenbeschaffenheit - Messung von Enzymaktivitätsmustern in Bodenproben mit kolorimetrischen Substraten in Mikrotiterplatten (ISO 20130:2018)

Dieses Dokument legt ein Verfahren zur gemeinsamen (oder getrennten) Messung verschiedener Hydrolase-Aktivitäten (Arylamidase, Arylsulfatase, β Galactosidase, α Glucosidase, β Glucosidase, N Acetyl-Glucosa¬minidase, saure, alkalische und globale Phosphatasen, Urease) in Bodenproben unter Verwendung von kolorimetrischen Substraten fest. Die Enzymaktivitäten im Boden variieren je nach Jahreszeit und sind abhängig von chemischen, physikalischen und biologischen Eigenschaften des Bodens.
Diese Methode kann entweder zum Nachweis schädlicher Auswirkungen auf die Enzymaktivitäten im Boden durch toxische Substanzen oder andere anthropogene Stoffe in kontaminierten Böden gegenüber einem Kontrollboden oder zur Prüfung von Chemikalien angewendet werden.

Qualité du sol - Mesure de l'activité enzymatique dans des échantillons de sol en utilisant des substrats colorimétriques (ISO 20130:2018)

Le présent document spécifie une méthode de mesure simultanée (ou non) de plusieurs activités des hydrolases (arylamidase, arylsulfatase, β-galactosidase, α-glucosidase, β-glucosidase, N-acétylglucosaminidase, phosphatases acides, alcalines et globales, uréase) dans des échantillons de sol en utilisant des substrats colorimétriques. Les activités enzymatiques du sol varient en fonction des saisons et dépendent des caractéristiques chimiques, physiques et biologiques du sol. Cette méthode peut être appliquée soit pour la détection des effets nocifs de substances toxiques ou d'autres agents anthropiques dans un sol contaminé par comparaison avec un sol de référence, soit pour la réalisation d'essais sur des produits chimiques.

Kakovost tal - Merjenje encimske aktivnosti v vzorcih tal s kolorometričnimi substrati na mikrotitrskih ploščah (ISO 20130:2018)

General Information

Status
Published
Publication Date
05-May-2020
Withdrawal Date
29-Nov-2020
Current Stage
6060 - Definitive text made available (DAV) - Publishing
Start Date
06-May-2020
Due Date
07-Mar-2022
Completion Date
06-May-2020

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SLOVENSKI STANDARD
01-november-2020
Kakovost tal - Merjenje encimske aktivnosti v vzorcih tal s kolorometričnimi
substrati na mikrotitrskih ploščah (ISO 20130:2018)
Soil quality - Measurement of enzyme activity patterns in soil samples using colorimetric
substrates in micro-well plates (ISO 20130:2018)
Bodenbeschaffenheit - Messung von Enzymaktivitätsmustern in Bodenproben mit
kolorimetrischen Substraten in Mikrotiterplatten (ISO 20130:2018)
Qualité du sol - Mesure de l'activité enzymatique dans des échantillons de sol en utilisant
des substrats colorimétriques (ISO 20130:2018)
Ta slovenski standard je istoveten z: EN ISO 20130:2020
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 20130
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2020
EUROPÄISCHE NORM
ICS 13.080.30
English Version
Soil quality - Measurement of enzyme activity patterns in
soil samples using colorimetric substrates in micro-well
plates (ISO 20130:2018)
Qualité du sol - Mesure de l'activité enzymatique dans Bodenbeschaffenheit - Messung von
des échantillons de sol en utilisant des substrats Enzymaktivitätsmustern in Bodenproben mit
colorimétriques (ISO 20130:2018) kolorimetrischen Substraten in Mikrotiterplatten (ISO
20130:2018)
This European Standard was approved by CEN on 13 April 2020.

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, Turkey 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
© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20130:2020 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
The text of ISO 20130:2018 has been prepared by Technical Committee ISO/TC 190 "Soil quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 20130:2020 by
Technical Committee CEN/TC 444 “Environmental characterization of solid matrices” the secretariat of
which is held by NEN.
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 November 2020, and conflicting national standards
shall be withdrawn at the latest by November 2020.
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.
According to the CEN-CENELEC Internal Regulations, the national standards organizations 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, Turkey and the
United Kingdom.
Endorsement notice
The text of ISO 20130:2018 has been approved by CEN as EN ISO 20130:2020 without any modification.

INTERNATIONAL ISO
STANDARD 20130
First edition
2018-07
Soil quality — Measurement of
enzyme activity patterns in soil
samples using colorimetric substrates
in micro-well plates
Qualité du sol — Mesure de l'activité enzymatique dans des
échantillons de sol en utilisant des substrats colorimétriques
Reference number
ISO 20130:2018(E)
©
ISO 2018
ISO 20130:2018(E)
© ISO 2018
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting
on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address
below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2018 – All rights reserved

ISO 20130:2018(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions, symbols and abbreviated terms . 1
3.1 Terms and definitions . 1
3.2 Symbols and abbreviated terms. 1
4 Principle . 2
5 Reactives . 2
5.1 Buffers and reageants . 2
5.2 Substrates and standards . 5
5.2.1 Preparation of standard solutions . 5
5.2.2 Preparation of substrate solutions . 5
6 Apparatus and materials. 7
7 Procedure. 8
7.1 Establishment of calibration curves . 8
7.1.1 General. 8
7.1.2 Solution of PNP . . 8
7.1.3 Solution of β-naphthylamine . 8
7.1.4 Solution of ammonium chloride . 8
7.2 Sampling . 9
7.2.1 Sample preparation . 9
7.2.2 Addition of substrate .10
7.2.3 Absorbance measurements .11
7.2.4 Measurements of urease activities .12
8 Calculation of results .12
9 Expression of results .12
10 Validity criteria .12
11 Interlaboratory validation .13
12 Test report .13
Annex A (informative) Validation of the method and intralaboratory tests for evaluating
soil enzymatic activities with colorimetric method .14
Annex B (informative) International ring test for evaluating soil enzymatic activities
with colorimetric method .19
Bibliography .28
ISO 20130:2018(E)
Foreword
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