Determination of the ultimate aerobic biodegradability of plastic materials in an aqueous medium - Method by analysis of evolved carbon dioxide (ISO 14852:2021)

This document specifies a method, by measuring the amount of carbon dioxide evolved, for the
determination of the degree of aerobic biodegradability of plastic materials, including those containing
formulation additives. The test material is exposed in a synthetic medium under standardized
laboratory conditions to an inoculum from activated sludge under aerobic conditions.
The conditions used in this document do not necessarily correspond to the optimum conditions allowing
maximum biodegradation to occur, but this test method is designed to measure the biodegradation of
plastic materials and give an indication of their potential biodegradability.
The method enables the assessment of the biodegradation to be improved by calculating a carbon
balance (optional, see Annex C).
The method applies to the following materials:
— natural and/or synthetic polymers, copolymers or mixtures thereof;
— plastic materials which contain additives such as plasticizers, colorants or other compounds;
— water-soluble polymers;
— materials which, under the test conditions, do not inhibit the microorganisms present in the
inoculum. Inhibitory effects can be determined using an inhibition control or by another appropriate
method (see, for example, ISO 8192[1]). If the test material is inhibitory to the inoculum, a lower test
concentration, another inoculum or a pre-exposed inoculum can be used.

Bestimmung der vollständigen aeroben Bioabbaubarkeit von Kunststoff-Materialien in einem wässrigen Medium - Verfahren mittels Analyse des freigesetzten Kohlenstoffdioxides (ISO 14852:2021)

Dieses Dokument legt ein Verfahren fest, mit dem durch Messung der gebildeten Menge an Kohlenstoffdioxid der Grad der aeroben Bioabbaubarkeit von Kunststoffen, einschließlich solcher Kunststoffe, die Formulierungs-Additive enthalten, bestimmt werden kann. Die Prüfsubstanz wird in einem synthetischen Medium unter standardisierten Laborbedingungen einem Inokulum aus Belebtschlamm, ausgereiftem Kompost oder Boden unter aeroben, mesophilen Bedingungen ausgesetzt.
Wird ein nicht adaptierter Belebtschlamm als Inokulum verwendet, kann das Prüfergebnis zur Beurteilung der aeroben Bioabbauvorgänge verwendet werden, die in der Umgebung einer Kläranlage auftreten. Wird ein gemischtes oder voradaptiertes Inokulum verwendet, kann das Verfahren dazu dienen, die potentielle Bioabbaubarkeit einer Prüfsubstanz zu untersuchen.
Die in diesem Dokument angewendeten Bedingungen entsprechen nicht notwendigerweise den optimalen Bedingungen, die das Auftreten eines maximalen Bioabbaus zulassen; aber dieses Prüfverfahren ist für die Messung der Bioabbaubarkeit von Kunststoffen und dafür vorgesehen, Erkenntnisse hinsichtlich deren potentieller Bioabbaubarkeit zu liefern.
Das Verfahren lässt durch die Berechnung einer Kohlenstoffbilanz (wahlweise siehe Anhang C) die Verbesserung der Beurteilung der Bioabbaubarkeit zu.
Das Verfahren ist auf folgende Substanzen anwendbar:
- natürliche und/oder synthetische Polymere, Copolymere oder Gemische aus diesen;
- Kunststoffe, die Additive, wie z. B. Weichmacher, Farbstoffe oder andere Verbindungen, enthalten;
- wasserlösliche Polymere;
- Substanzen, die unter Prüfbedingungen die im Inokulum vorhandenen Mikroorganismen nicht inhibieren. Inhibierungswirkungen lassen sich durch eine Inhibitionskontrolle oder ein anderes geeignetes Verfahren bestimmen (siehe z. B. ISO 8192 [1]). Wirkt die Prüfsubstanz inhibierend auf das Inokulum, kann eine geringere Prüfkonzentration, ein anderes oder ein voradaptiertes Inokulum verwendet werden.

Évaluation de la biodégradabilité aérobie ultime des matériaux plastiques en milieu aqueux - Méthode par analyse du dioxyde de carbone libéré (ISO 14852:2021)

Določanje končne aerobne biorazgradljivosti polimernih materialov v vodnem mediju - Metoda z analizo sproščenega ogljikovega dioksida (ISO 14852:2021)

General Information

Status
Published
Public Enquiry End Date
23-Sep-2020
Publication Date
21-Jul-2021
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
21-Jul-2021
Due Date
25-Sep-2021
Completion Date
22-Jul-2021

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SLOVENSKI STANDARD
SIST EN ISO 14852:2021
01-september-2021
Nadomešča:
SIST EN ISO 14852:2018
Določanje končne aerobne biorazgradljivosti polimernih materialov v vodnem
mediju - Metoda z analizo sproščenega ogljikovega dioksida (ISO 14852:2021)

Determination of the ultimate aerobic biodegradability of plastic materials in an aqueous

medium - Method by analysis of evolved carbon dioxide (ISO 14852:2021)

Bestimmung der vollständigen aeroben Bioabbaubarkeit von Kunststoff-Materialien in

einem wässrigen Medium - Verfahren mittels Analyse des freigesetzten
Kohlenstoffdioxides (ISO 14852:2021)

Évaluation de la biodégradabilité aérobie ultime des matériaux plastiques en milieu

aqueux - Méthode par analyse du dioxyde de carbone libéré (ISO 14852:2021)
Ta slovenski standard je istoveten z: EN ISO 14852:2021
ICS:
83.080.01 Polimerni materiali na Plastics in general
splošno
SIST EN ISO 14852:2021 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 14852:2021
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SIST EN ISO 14852:2021
EN ISO 14852
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2021
EUROPÄISCHE NORM
ICS 83.080.01 Supersedes EN ISO 14852:2018
English Version
Determination of the ultimate aerobic biodegradability of
plastic materials in an aqueous medium - Method by
analysis of evolved carbon dioxide (ISO 14852:2021)

Évaluation de la biodégradabilité aérobie ultime des Bestimmung der vollständigen aeroben

matériaux plastiques en milieu aqueux - Méthode par Bioabbaubarkeit von Kunststoff-Materialien in einem

analyse du dioxyde de carbone libéré (ISO wässrigen Medium - Verfahren mittels Analyse des

14852:2021) freigesetzten Kohlenstoffdioxides (ISO 14852:2021)
This European Standard was approved by CEN on 19 June 2021.

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

© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 14852:2021 E

worldwide for CEN national Members.
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SIST EN ISO 14852:2021
EN ISO 14852:2021 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

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SIST EN ISO 14852:2021
EN ISO 14852:2021 (E)
European foreword

This document (EN ISO 14852:2021) has been prepared by Technical Committee ISO/TC 61 "Plastics"

in collaboration with Technical Committee CEN/TC 249 “Plastics” the secretariat of which is held by

NBN.

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 2022, and conflicting national standards shall

be withdrawn at the latest by January 2022.

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 EN ISO 14852:2018.

Any feedback and questions on this document should be directed to the users’ national standards

body/national committee. A complete listing of these bodies can be found on the CEN websites.

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 14852:2021 has been approved by CEN as EN ISO 14852:2021 without any modification.

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SIST EN ISO 14852:2021
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SIST EN ISO 14852:2021
INTERNATIONAL ISO
STANDARD 14852
Third edition
2021-06
Determination of the ultimate aerobic
biodegradability of plastic materials
in an aqueous medium — Method by
analysis of evolved carbon dioxide
Évaluation de la biodégradabilité aérobie ultime des matériaux
plastiques en milieu aqueux — Méthode par analyse du dioxyde de
carbone libéré
Reference number
ISO 14852:2021(E)
ISO 2021
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2021

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
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2021 – All rights reserved
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Principle ........................................................................................................................................................................................................................ 3

5 Test environment ................................................................................................................................................................................................. 4

6 Reagents ........................................................................................................................................................................................................................ 4

7 Apparatus ..................................................................................................................................................................................................................... 6

8 Procedure..................................................................................................................................................................................................................... 7

8.1 Test material ............................................................................................................................................................................................. 7

8.2 Reference material ............................................................................................................................................................................... 7

8.3 Preparation of the inoculum ....................................................................................................................................................... 7

8.3.1 General...................................................................................................................................................................................... 7

8.3.2 Inoculum from wastewater-treatment plants ........................................................................................ 8

8.4 Test .................................................................................................................................................................................................................... 8

9 Calculation and expression of results ..........................................................................................................................................10

9.1 Calculation ...............................................................................................................................................................................................10

9.1.1 Theoretical amount of carbon dioxide evolved by the test material................................10

9.1.2 Percentage biodegradation from CO evolution ................................................................................10

9.2 Expression and interpretation of results ......................................................................................................................10

10 Validity of results ..............................................................................................................................................................................................11

11 Test report ................................................................................................................................................................................................................11

Annex A (informative) Principle of a system for measuring evolved carbon dioxide (example) .......13

Annex B (informative) Examples of methods for the determination of evolved carbon dioxide .......14

Annex C (informative) Example of the determination of a carbon balance ..............................................................16

Annex D (informative) Example of a determination of the amount of water insoluble

polymer remaining at the end of a biodegradation test and the molecular mass of

the polymer .............................................................................................................................................................................................................18

Bibliography .............................................................................................................................................................................................................................19

© ISO 2021 – All rights reserved iii
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).

Attention is drawn to the possibility that some of the elements of this document can be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www .iso .org/ patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www .iso .org/

iso/ foreword .html.

This document was prepared by Technical Committee ISO/TC 61, Plastics, Subcommittee SC 14,

Environmental aspects, in collaboration with the European Committee for Standardization (CEN)

Technical Committee CEN/TC 249, Plastics, in accordance with the Agreement on technical cooperation

between ISO and CEN (Vienna Agreement).

This third edition cancels and replaces the second edition (ISO 14852:2018), which has been technically

revised. The main changes compared to the previous edition are as follows:

— in the Scope and Clause 8, soil and compost have been excluded for the inoculums used in this

document;

— in 8.4, number of flasks for checking the inoculum activity have been changed from three to two;

— the validity criteria has been revised to conform with ISO 14851.

Any feedback or questions on this document should be directed to the user’s national standards body. A

complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2021 – All rights reserved
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
Introduction

With the increasing use of plastics, their recovery and disposal have become a major issue. As a first

priority, recovery should be promoted. Biodegradable plastics are now emerging as one of the options

available to solve such environmental problems. Plastic materials, such as products or packaging, which

are sent to composting facilities should be potentially biodegradable. Therefore, it is very important to

determine the potential biodegradability of such materials and to obtain an indication of their potential

biodegradability.
© ISO 2021 – All rights reserved v
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SIST EN ISO 14852:2021
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SIST EN ISO 14852:2021
INTERNATIONAL STANDARD ISO 14852:2021(E)
Determination of the ultimate aerobic biodegradability
of plastic materials in an aqueous medium — Method by
analysis of evolved carbon dioxide

WARNING — Sewage, activated sludge can contain potentially pathogenic organisms. Therefore,

appropriate precautions should be taken when handling them. Toxic test compounds and those

whose properties are unknown should be handled with care.
1 Scope

This document specifies a method, by measuring the amount of carbon dioxide evolved, for the

determination of the degree of aerobic biodegradability of plastic materials, including those containing

formulation additives. The test material is exposed in a synthetic medium under standardized

laboratory conditions to an inoculum from activated sludge under aerobic conditions.

The conditions used in this document do not necessarily correspond to the optimum conditions allowing

maximum biodegradation to occur, but this test method is designed to measure the biodegradation of

plastic materials and give an indication of their potential biodegradability.

The method enables the assessment of the biodegradation to be improved by calculating a carbon

balance (optional, see Annex C).
The method applies to the following materials:
— natural and/or synthetic polymers, copolymers or mixtures thereof;

— plastic materials which contain additives such as plasticizers, colorants or other compounds;

— water-soluble polymers;

— materials which, under the test conditions, do not inhibit the microorganisms present in the

inoculum. Inhibitory effects can be determined using an inhibition control or by another appropriate

[1]

method (see, for example, ISO 8192 ). If the test material is inhibitory to the inoculum, a lower test

concentration, another inoculum or a pre-exposed inoculum can be used.
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.

ISO 8245, Water quality — Guidelines for the determination of total organic carbon (TOC) and dissolved

organic carbon (DOC)
3 Terms and definitions
For the purposes of this document, the following terms and definitions 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/
© ISO 2021 – All rights reserved 1
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
3.1
ultimate aerobic biodegradability

breakdown ratio by expressed as percentage of an organic compound by microorganisms in the presence

of oxygen into carbon dioxide, water and mineral salts of any other elements present (mineralization)

plus new biomass
3.2
activated sludge

mixture of organic materials and biomass produced in the aerobic treatment of waste water by the

growth of bacteria and other microorganisms in the presence of dissolved oxygen
3.3
concentration of suspended solids

amount of solids obtained by filtration or centrifugation of a known volume of activated sludge (3.2)

and drying at about 105 °C to constant mass
3.4
dissolved inorganic carbon
DIC

part of the inorganic carbon in water which cannot be removed by specified phase separation

Note 1 to entry: Phase separation can be achieved for example by centrifugation at 40 000 m⋅s for 15 min or by

membrane filtration using membranes with pores of 0,2 µm to 0,45 µm diameter.
3.5
theoretical amount of evolved carbon dioxide
ThCO

maximum theoretical amount of carbon dioxide evolved after completely oxidizing a chemical

compound, calculated from the molecular formula
Note 1 to entry: It is calculated from the molecular formula.

Note 2 to entry: It is expressed as milligrams of carbon dioxide evolved per milligram or gram of test compound.

3.6
total organic carbon
TOC
amount of carbon bound in an organic compound

Note 1 to entry: It is expressed as milligrams of carbon per 100 mg of the compound.

[SOURCE: ISO 17556:2012, 3.14]
3.7
dissolved organic carbon
DOC

part of the organic carbon in water which cannot be removed by specified phase separation

Note 1 to entry: Phase separation can be achieved for example by centrifugation at 40 000 m⋅s for 15 min or by

membrane filtration using membranes with pores of 0,2 µm to 0,45 µm diameter.
3.8
lag phase

time from the start of a test until adaptation and/or selection of the degrading microorganisms is

achieved and the degree of biodegradation of a chemical compound or organic matter has increased to

about 10 % of the maximum level of biodegradation (3.9)
Note 1 to entry: It is measured in days.
2 © ISO 2021 – All rights reserved
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
3.9
maximum level of biodegradation

degree of biodegradation of a chemical compound or organic matter in a test, above which no further

biodegradation takes place during the test
Note 1 to entry: It is measured in per cent.
3.10
biodegradation phase

time from the end of the lag phase (3.8) of a test until the plateau phase (3.11) has been reached

Note 1 to entry: It is measured in days.
3.11
plateau phase
time from the end of the biodegradation phase (3.10) until the end of a test
Note 1 to entry: It is measured in days.
3.12
pre-exposure

pre-incubation of an inoculum (3.14) in the presence of the chemical compound or organic matter under

test, with the aim of enhancing the ability of the inoculum to biodegrade the test material by adaptation

and/or selection of the microorganisms
3.13
pre-conditioned

pre-incubated inoculum (3.14) under the conditions of the subsequent test in the absence of the chemical

compound or organic matter under test, with the aim of improving the test by acclimatization of the

microorganisms to the test conditions
3.14
inoculum
microorganisms or other material used in an inoculation (3.15)
Note 1 to entry: Also called inoculant.
3.15
inoculation

introduction of microorganisms into a culture medium in order to start a biological process

4 Principle

The biodegradability of a plastic material is determined using aerobic, mesophilic microorganisms

in an aqueous system. The test mixture contains an inorganic medium, the organic test material (the

sole source of carbon and energy) with a concentration between 100 mg/l and 2 000 mg/l of organic

carbon and activated sludge as the inoculum. If higher concentrations of test material are used, then an

optimised test medium should be applied.

NOTE Lower concentrations such as those between 20 mg/l and 40 mg/l of organic carbon have been tested

and found suitable.

The mixture is agitated in test flasks and aerated with carbon-dioxide-free air over a period of time

depending on the biodegradation kinetics, but not exceeding 2 months. The carbon dioxide evolved

during the microbial degradation is determined by a suitable analytical method, examples of which are

given in Annexes A and B.

The level of biodegradation is determined by comparing the amount of carbon dioxide evolved with

the theoretical amount (ThCO ) and expressed in per cent. The test result is the maximum level of

biodegradation, determined from the plateau phase of the biodegradation curve. Optionally, a carbon

balance can be calculated to give additional information on the biodegradation (see Annex C).

© ISO 2021 – All rights reserved 3
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SIST EN ISO 14852:2021
ISO 14852:2021(E)

Unlike ISO 9439, which is used for a variety of organic compounds, this document is specially designed

for the determination of the biodegradation of plastic materials. The special requirements necessary

affect the choice of the inoculum and the test medium, and there is the possibility of improving the

evaluation of the biodegradability by calculating a carbon balance.
5 Test environment

Incubation shall take place in the dark or in diffuse light in an enclosure which is free from vapours

inhibitory to microorganisms and which is maintained at a constant temperature, preferably between

20 °C and 25 °C, to an accuracy of ±1 °C.
6 Reagents
Use only reagents of recognized analytical grade.

6.1 Distilled or deionized water, free of toxic substances (copper in particular) and containing less

than 2 mg/l of DOC.
6.2 Test medium.

Depending on the purpose of the test, different test media can be used. For example, if a test material

is used at higher concentrations, use the optimized test medium (6.2.2) with higher buffering capacity

and nutrient concentrations.
6.2.1 Standard test medium.
6.2.1.1 Solution A.
Dissolve the following in water (6.1) and make up to 1 000 ml.
anhydrous potassium dihydrogen phosphate (KH PO ) 8,5 g
2 4
anhydrous dipotassium hydrogen phosphate (K HPO ) 21,75 g
2 4
disodium hydrogen phosphate dihydrate (Na HPO ⋅ 2H O) 33,4 g
2 4 2
ammonium chloride (NH Cl) 0,5 g

The correct composition of the solution can be checked by measuring the pH, which should be 7,4.

6.2.1.2 Solution B.

Dissolve 22,5 g of magnesium sulfate heptahydrate (MgSO ⋅ 7H O) in water (6.1) and make up to

4 2
1 000 ml.
6.2.1.3 Solution C.

Dissolve 36,4 g of calcium chloride dihydrate (CaCl ⋅ 2H O) in water (6.1) and make up to 1 000 ml.

2 2
6.2.1.4 Solution D.

Dissolve 0,25 g of iron(III) chloride hexahydrate (FeCl ⋅ 6H O) in water (6.1) and make up to 1 000 ml.

3 2

Prepare this solution freshly before use to avoid precipitation, or add a drop of concentrated

hydrochloric acid (HCl) or a drop of 0,4 g/l aqueous solution of ethylenediaminetetraacetic acid (EDTA).

4 © ISO 2021 – All rights reserved
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SIST EN ISO 14852:2021
ISO 14852:2021(E)
6.2.1.5 Preparation.

To prepare 1 l of test medium, add the following, to about 500 ml of water (6.1):

— 10 ml of solution A (6.2.1.1);
— 1 ml of each of solutions B (6.2.1.2), C (6.2.1.3), D (6.2.1.4).
Make up to 1 000 ml with water (6.1).

Prepare the test medium freshly before use. The solutions A up to C can be stored up to 6 months in the

dark at room temperature. The same applies for solution D in case HCl or EDTA has been added.

6.2.2 Optimized test medium.

This optimized medium is highly buffered and contains more inorganic nutrients. This is necessary to

keep the pH constant in the system during the test, even at high concentrations of the test material. The

medium contains about 2 400 mg/l of phosphorus and 50 mg/l of nitrogen and is therefore suitable

for concentrations in the test material of up to 2 000 mg/l of organic carbon. If higher or lower test-

material concentrations are used, increase or decrease respectively the nitrogen content to keep the

C:N ratio at about 40:1.
6.2.2.1 Solution E.
Dissolve the following in water (6.1) and make up to 1 000 ml.
anhydrous potassium dihydrogen phosphate (KH PO ) 37,5 g
2 4
disodium hydrogen phosphate dihydrate (Na HPO ⋅ 2H O) 87,3 g
2 4 2
ammonium chloride (NH Cl) 2,0 g
6.2.2.2 Solution F (trace-element solution, optional).

Dissolve in 10 ml of aqueous HCl solution (25 %, 7,7 mol/l), in the following sequence:

a) 70 mg of ZnCl ;
b) 100 mg of MnCl ⋅ 4H O;
2 2
c) 6 mg of H BO ;
3 3
d) 190 mg of CoCl ⋅ 6H O;
2 2
e) 3 mg of CuCl ⋅ 2H O;
2 2
f) 240 mg of NiCl ⋅ 6H O;
2 2
g) 36 mg of Na MoO ⋅ 2H O;
2 4 2
h) 33 mg of Na WO ⋅ 2H O;
2 4 2
i) 26 mg of Na SeO ⋅ 5H O.
2 3 2
Make up to 1 000 ml with water (6.1).
6.2.2.3 Solution G (vitamin solution, optional).

Dissolve in 100 ml of water (6.1) 0,6 mg of biotine, 2,0 mg of niacinamide, 2,0 mg of p-aminobenzoate,

1,0 mg of panthotenic acid, 10,0 mg of pyridoxal hydrochloride, 5,0 mg of cyanocobalamine, 2,0 mg of

folic acid, 5,0 mg of riboflavin, 5,0 mg of DL-thioctic acid and 1,0 mg of thiamine dichloride or use a

© ISO 2021 – All rights reserved 5
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SIST EN ISO 14852:2021
ISO 14852:2021(E)

solution of 15 mg of yeast extract in 100 ml of water (6.1). Filter the solution for sterilization using

membrane filters (see 7.7).

Solutions F and G are optional and are not required if a sufficient concentration of the inoculum is used,

such as activated sludge. It is recommended that 1 ml portions be prepared and kept refrigerated until

use.
6.2.2.4 Preparation.
To prepare 1 l of test medium, add, to about 800 ml of water (6.1)
— 100 ml of solution E (6.2.2.1), and

— 1 ml of each of solutions B (6.2.1.2), C (6.2.1.3), D (6.2.1.4) and, optionally, F (6.2.2.2) and G (6.2.2.3).

Make up to 1 000 ml with water (6.1) and measure the pH.

The correct composition of the test medium can be checked by measuring the pH, which should be

7,0 ± 0,2.
7 Apparatus

7.1 Ensure that all glassware is thoroughly cleaned and, in particular, free from organic or toxic matter.

Usual laboratory equipment are required, plus the following.

7.2 Test flasks, glass vessels (e.g. bottles or conical flasks) designed to allow gas purging and shaking

or stirring, and fitted with tubing impermeable to CO . The vessels shall be located in a constant-

temperature room or in a thermostatted apparatus (e.g. water-bath).

7.3 CO -free-air production system, capable of supplying CO -free air at a flow rate between 50 ml/

2 2

min and 100 ml/min to each test flask, held constant to within ±10 % (see example of system, including

test vessels, in Annex A).

7.4 Analytical instrument for determining carbon dioxide, consisting of any suitable ap

...

SLOVENSKI STANDARD
oSIST prEN ISO 14852:2020
01-september-2020
Določanje končne aerobne biorazgradljivosti polimernih materialov v vodnem
mediju - Metoda z analizo sproščenega ogljikovega dioksida (ISO/DIS 14852:2020)

Determination of the ultimate aerobic biodegradability of plastic materials in an aqueous

medium - Method by analysis of evolved carbon dioxide (ISO/DIS 14852:2020)

Bestimmung der vollständigen aeroben Bioabbaubarkeit von Kunststoff-Materialien in

einem wässrigen Medium - Verfahren mittels Analyse des freigesetzten
Kohlenstoffdioxides (ISO/DIS 14852:2020)

Évaluation de la biodégradabilité aérobie ultime des matériaux plastiques en milieu

aqueux - Méthode par analyse du dioxyde de carbone libéré (ISO/DIS 14852:2020)
Ta slovenski standard je istoveten z: prEN ISO 14852
ICS:
83.080.01 Polimerni materiali na Plastics in general
splošno
oSIST prEN ISO 14852:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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oSIST prEN ISO 14852:2020
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oSIST prEN ISO 14852:2020
DRAFT INTERNATIONAL STANDARD
ISO/DIS 14852
ISO/TC 61/SC 14 Secretariat: DIN
Voting begins on: Voting terminates on:
2020-07-02 2020-09-24
Determination of the ultimate aerobic biodegradability
of plastic materials in an aqueous medium — Method by
analysis of evolved carbon dioxide

Évaluation de la biodégradabilité aérobie ultime des matériaux plastiques en milieu aqueux — Méthode

par analyse du dioxyde de carbone libéré
ICS: 83.080.01
THIS DOCUMENT IS A DRAFT CIRCULATED
This document is circulated as received from the committee secretariat.
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
NOT BE REFERRED TO AS AN INTERNATIONAL
STANDARD UNTIL PUBLISHED AS SUCH.
IN ADDITION TO THEIR EVALUATION AS
ISO/CEN PARALLEL PROCESSING
BEING ACCEPTABLE FOR INDUSTRIAL,
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
Reference number
NATIONAL REGULATIONS.
ISO/DIS 14852:2020(E)
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. ISO 2020
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oSIST prEN ISO 14852:2020
ISO/DIS 14852:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020

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

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below or ISO’s member body in the country of the requester.
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Published in Switzerland
ii © ISO 2020 – All rights reserved
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oSIST prEN ISO 14852:2020
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Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Principle ........................................................................................................................................................................................................................ 3

5 Test environment ................................................................................................................................................................................................. 4

6 Reagents ........................................................................................................................................................................................................................ 4

7 Apparatus ..................................................................................................................................................................................................................... 6

8 Procedure..................................................................................................................................................................................................................... 7

8.1 Test material ............................................................................................................................................................................................. 7

8.2 Reference material ............................................................................................................................................................................... 7

8.3 Preparation of the inoculum ....................................................................................................................................................... 7

8.3.1 General...................................................................................................................................................................................... 7

8.3.2 Inoculum from wastewater-treatment plants ........................................................................................ 8

8.4 Test .................................................................................................................................................................................................................... 8

9 Calculation and expression of results ............................................................................................................................................. 9

9.1 Calculation .................................................................................................................................................................................................. 9

9.1.1 Theoretical amount of carbon dioxide evolved by the test material................................... 9

9.1.2 Percentage biodegradation from CO evolution ................................................................................10

9.2 Expression and interpretation of results ......................................................................................................................10

10 Validity of results ..............................................................................................................................................................................................11

11 Test report ................................................................................................................................................................................................................11

Annex A (informative) Principle of a system for measuring evolved carbon dioxide (example) .......13

Annex B (informative) Examples of methods for the determination of evolved carbon dioxide .......14

Annex C (informative) Example of the determination of a carbon balance ..............................................................16

Annex D (informative) Example of a determination of the amount of water insoluble

polymer remaining at the end of a biodegradation test and the molecular mass of

the polymer .............................................................................................................................................................................................................18

Bibliography .............................................................................................................................................................................................................................19

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Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www .iso .org/ patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www .iso .org/

iso/ foreword .html.

This document was prepared by Technical Committee ISO/TC 61, Plastics, Subcommittee SC 14,

Environmental aspects.

This third edition cancels and replaces the second edition (ISO 14852:2018), which has been technically

revised. The main changes compared to the previous edition are as follows:

— in Scope and Clause 8, soil and compost have been excluded for the inoculums used in this document;

— in 8.4, number of flask for checking the inoculum activity have been changed from three to two

— the validity criteria has been revised to comply with ISO 14851;

Any feedback or questions on this document should be directed to the user’s national standards body. A

complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2020 – All rights reserved
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Introduction

With the increasing use of plastics, their recovery and disposal have become a major issue. As a first

priority, recovery should be promoted. Biodegradable plastics are now emerging as one of the options

available to solve such environmental problems. Plastic materials, such as products or packaging, which

are sent to composting facilities should be potentially biodegradable. Therefore, it is very important to

determine the potential biodegradability of such materials and to obtain an indication of their potential

biodegradability.
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oSIST prEN ISO 14852:2020
DRAFT INTERNATIONAL STANDARD ISO/DIS 14852:2020(E)
Determination of the ultimate aerobic biodegradability
of plastic materials in an aqueous medium — Method by
analysis of evolved carbon dioxide

WARNING — Sewage, activated sludge may contain potentially pathogenic organisms. Therefore,

appropriate precautions should be taken when handling them. Toxic test compounds and those

whose properties are unknown should be handled with care.
1 Scope

This document specifies a method, by measuring the amount of carbon dioxide evolved, for the

determination of the degree of aerobic biodegradability of plastic materials, including those containing

formulation additives. The test material is exposed in a synthetic medium under standardized

laboratory conditions to an inoculum from activated sludge under aerobic, mesophilic conditions.

If an unadapted activated sludge is used as the inoculum, the test result can be used to assess the aerobic

biodegradation processes which occur in a waste water treatment plant environment. If a mixed or pre-

exposed inoculum is used, the method can be used to investigate the potential biodegradability of a test

material.

The conditions used in this document do not necessarily correspond to the optimum conditions allowing

maximum biodegradation to occur, but this test method is designed to measure the biodegradation of

plastic materials and give an indication of their potential biodegradability.

The method enables the assessment of the biodegradation to be improved by calculating a carbon

balance (optional, see Annex C).
The method applies to the following materials:
— natural and/or synthetic polymers, copolymers or mixtures thereof;

— plastic materials which contain additives such as plasticizers, colorants or other compounds;

— water-soluble polymers;

— materials which, under the test conditions, do not inhibit the microorganisms present in the

inoculum. Inhibitory effects can be determined using an inhibition control or by another appropriate

[1]

method (see, for example, ISO 8192 ). If the test material is inhibitory to the inoculum, a lower test

concentration, another inoculum or a pre-exposed inoculum can be used.
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.

ISO 8245, Water quality — Guidelines for the determination of total organic carbon (TOC) and dissolved

organic carbon (DOC)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
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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/
3.1
ultimate aerobic biodegradation

breakdown of an organic compound by microorganisms in the presence of oxygen into carbon dioxide,

water and mineral salts of any other elements present (mineralization) plus new biomass

3.2
activated sludge

mixture of organic materials and biomass produced in the aerobic treatment of waste water by the

growth of bacteria and other microorganisms in the presence of dissolved oxygen
3.3
concentration of suspended solids in an activated sludge

amount of solids obtained by filtration or centrifugation of a known volume of activated sludge and

drying at about 105 °C to constant mass
3.4
dissolved inorganic carbon
DIC

part of the inorganic carbon in water which cannot be removed by specified phase separation

Note 1 to entry: Phase separation can be achieved for example by centrifugation at 40 000 m⋅s for 15 min or by

membrane filtration using membranes with pores of 0,2 µm to 0,45 µm diameter.
3.5
theoretical amount of evolved carbon dioxide
ThCO

maximum theoretical amount of carbon dioxide evolved after completely oxidizing a chemical

compound, calculated from the molecular formula

Note 1 to entry: It is expressed as milligrams of carbon dioxide evolved per milligram or gram of test compound.

3.6
total organic carbon
TOC
amount of carbon bound in an organic compound

Note 1 to entry: It is expressed as milligrams of carbon per 100 mg of the compound.

[SOURCE: ISO 17556:2012, 3.14]
3.7
dissolved organic carbon
DOC

part of the organic carbon in water which cannot be removed by specified phase separation

Note 1 to entry: Phase separation can be achieved for example by centrifugation at 40 000 m⋅s for 15 min or by

membrane filtration using membranes with pores of 0,2 µm to 0,45 µm diameter.
3.8
lag phase

time from the start of a test until adaptation and/or selection of the degrading microorganisms is

achieved and the degree of biodegradation of a chemical compound or organic matter has increased to

about 10 % of the maximum level of biodegradation
Note 1 to entry: It is measured in days.
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3.9
maximum level of biodegradation

degree of biodegradation of a chemical compound or organic matter in a test, above which no further

biodegradation takes place during the test
Note 1 to entry: It is measured in per cent.
3.10
biodegradation phase

time from the end of the lag phase of a test until the plateau phase has been reached

Note 1 to entry: It is measured in days.
3.11
plateau phase
time from the end of the biodegradation phase until the end of a test
Note 1 to entry: It is measured in days.
3.12
pre-exposure

pre-incubation of an inoculum in the presence of the chemical compound or organic matter under test,

with the aim of enhancing the ability of the inoculum to biodegrade the test material by adaptation

and/or selection of the microorganisms
3.13
pre-conditioning

pre-incubation of an inoculum under the conditions of the subsequent test in the absence of the chemical

compound or organic matter under test, with the aim of improving the test by acclimatization of the

microorganisms to the test conditions
3.14
inoculum
microorganisms or other material used in an inoculation
Note 1 to entry: Also called inoculant.
3.15
inoculation

introduction of microorganisms into a culture medium in order to start a biological process

4 Principle

The biodegradability of a plastic material is determined using aerobic, mesophilic microorganisms

in an aqueous system. The test mixture contains an inorganic medium, the organic test material (the

sole source of carbon and energy) with a concentration between 100 mg/l and 2 000 mg/l of organic

carbon, and activated sludge as the inoculum. If higher concentrations of test material are used then an

optimised test medium should be applied.

NOTE Lower concentrations such as between 20 mg/l and 40 mg/l of organic carbon have been tested and

found suitable.

The mixture is agitated in test flasks and aerated with carbon-dioxide-free air over a period of time

depending on the biodegradation kinetics, but not exceeding 2 months. The carbon dioxide evolved

during the microbial degradation is determined by a suitable analytical method, examples of which are

given in Annexes A and B.

The level of biodegradation is determined by comparing the amount of carbon dioxide evolved with

the theoretical amount (ThCO ) and expressed in per cent. The test result is the maximum level of

biodegradation, determined from the plateau phase of the biodegradation curve. Optionally, a carbon

balance may be calculated to give additional information on the biodegradation (see Annex C).

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Unlike ISO 9439, which is used for a variety of organic compounds, this document is specially designed

for the determination of the biodegradation of plastic materials. The special requirements necessary

affect the choice of the inoculum and the test medium, and there is the possibility of improving the

evaluation of the biodegradability by calculating a carbon balance.
5 Test environment

Incubation shall take place in the dark or in diffuse light in an enclosure which is free from vapours

inhibitory to microorganisms and which is maintained at a constant temperature, preferably between

20 °C and 25 °C, to an accuracy of ±1 °C.
6 Reagents
Use only reagents of recognized analytical grade.

6.1 Distilled or deionized water, free of toxic substances (copper in particular) and containing less

than 2 mg/l of DOC.
6.2 Test medium.

Depending on the purpose of the test, different test media may be used. For example, if a test material

is used at higher concentrations, use the optimized test medium (6.2.2) with higher buffering capacity

and nutrient concentrations.
6.2.1 Standard test medium.
6.2.1.1 Solution A.
Dissolve the following in water (6.1) and make up to 1 000 ml.
anhydrous potassium dihydrogen phosphate (KH PO ) 8,5 g
2 4
anhydrous dipotassium hydrogen phosphate (K HPO ) 21,75 g
2 4
disodium hydrogen phosphate dihydrate (Na HPO ⋅ 2H O) 33,4 g
2 4 2
ammonium chloride (NH Cl) 0,5 g

The correct composition of the solution can be checked by measuring the pH, which should be 7,4.

6.2.1.2 Solution B.

Dissolve 22,5 g of magnesium sulfate heptahydrate (MgSO ⋅ 7H O) in water (6.1) and make up to

4 2
1 000 ml.
6.2.1.3 Solution C.

Dissolve 36,4 g of calcium chloride dihydrate (CaCl ⋅ 2H O) in water (6.1) and make up to 1 000 ml.

2 2
6.2.1.4 Solution D.

Dissolve 0,25 g of iron(III) chloride hexahydrate (FeCl ⋅ 6H O) in water (6.1) and make up to 1 000 ml.

3 2

Prepare this solution freshly before use to avoid precipitation, or add a drop of concentrated

hydrochloric acid (HCl) or a drop of 0,4 g/l aqueous solution of ethylenediaminetetraacetic acid (EDTA).

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6.2.1.5 Preparation.

To prepare 1 l of test medium, add the following, to about 500 ml of water (6.1):

— 10 ml of solution A (6.2.1.1);
— 1 ml of each of solutions B (6.2.1.2), C (6.2.1.3), D (6.2.1.4).
Make up to 1 000 ml with water (6.1).

Prepare the test medium freshly before use. The solutions A up to C may be stored up to 6 months in the

dark at room temperature. The same applies for solution D in case HCl or EDTA has been added.

6.2.2 Optimized test medium.

This optimized medium is highly buffered and contains more inorganic nutrients. This is necessary to

keep the pH constant in the system during the test, even at high concentrations of the test material. The

medium contains about 2 400 mg/l of phosphorus and 50 mg/l of nitrogen and is therefore suitable

for concentrations in the test material of up to 2 000 mg/l of organic carbon. If higher or lower test-

material concentrations are used, increase or decrease respectively the nitrogen content to keep the

C:N ratio at about 40:1.
6.2.2.1 Solution E.
Dissolve the following in water (6.1) and make up to 1 000 ml.
anhydrous potassium dihydrogen phosphate (KH PO ) 37,5 g
2 4
disodium hydrogen phosphate dihydrate (Na HPO ⋅ 2H O) 87,3 g
2 4 2
ammonium chloride (NH Cl) 2,0 g
6.2.2.2 Solution F (trace-element solution, optional).

Dissolve in 10 ml of aqueous HCl solution (25 %, 7,7 mol/l), in the following sequence:

a) 70 mg of ZnCl ;
b) 100 mg of MnCl ⋅ 4H O;
2 2
c) 6 mg of H BO ;
3 3
d) 190 mg of CoCl ⋅ 6H O;
2 2
e) 3 mg of CuCl ⋅ 2H O;
2 2
f) 240 mg of NiCl ⋅ 6H O;
2 2
g) 36 mg of Na MoO ⋅ 2H O;
2 4 2
h) 33 mg of Na WO ⋅ 2H O;
2 4 2
i) 26 mg of Na SeO ⋅ 5H O.
2 3 2
Make up to 1 000 ml with water (6.1).
6.2.2.3 Solution G (vitamin solution, optional).

Dissolve in 100 ml of water (6.1) 0,6 mg of biotine, 2,0 mg of niacinamide, 2,0 mg of p-aminobenzoate,

1,0 mg of panthotenic acid, 10,0 mg of pyridoxal hydrochloride, 5,0 mg of cyanocobalamine, 2,0 mg of

folic acid, 5,0 mg of riboflavin, 5,0 mg of DL-thioctic acid and 1,0 mg of thiamine dichloride or use a

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solution of 15 mg of yeast extract in 100 ml of water (6.1). Filter the solution for sterilization using

membrane filters (see 7.6).

Solutions F and G are optional and are not required if a sufficient concentration of the inoculum is used,

e.g. activated sludge. It is recommended that 1 ml portions be prepared and kept refrigerated until use.

6.2.2.4 Preparation.
To prepare 1 litre of test medium, add, to about 800 ml of water (6.1),
— 100 ml of solution E (6.2.2.1), and

— 1 ml of each of solutions B (6.2.1.2), C (6.2.1.3), D (6.2.1.4) and, optionally, F (6.2.2.2) and G (6.2.2.3).

Make up to 1 000 ml with water (6.1) and measure the pH.

The correct composition of the test medium can be checked by measuring the pH, which should be

7,0 ± 0,2.
7 Apparatus

7.1 General. Ensure that all glassware is thoroughly cleaned and, in particular, free from organic or

toxic matter.
Required is usual laboratory equipment, plus the following.

7.2 Test flasks: glass vessels (e.g. bottles or conical flasks) designed to allow gas purging and shaking

or stirring, and fitted with tubing impermeable to CO . The vessels shall be located in a constant-

temperature room or in a thermostatted apparatus (e.g. water-bath).

7.3 CO -free-air production system, capable of supplying CO -free air at a flow rate between 50 ml/

2 2

min and 100 ml/min to each test flask, held constant to within ±10 % (see example of system, including

test vessels, in Annex A).

7.4 Analytical instrument for determining carbon dioxide, consisting of any suitable apparatus

with sufficient accuracy, e.g. a CO or DIC analyser or apparatus for titrimetric determination after

complete absorption in a basic solution (see examples in Annex B). Note that, if an analyser with an IR

detector, for instance, is used, CO -free air is not necessary.

7.5 Analytical equipment for measuring total organic carbon (TOC) and dissolved organic

carbon (DOC), see ISO 8245.
7.6 Analytical balance.

7.7 Centrifuge, or filtration device with membrane filters (0,45 µm pore size) which neither adsorb

nor release organic carbon significantly.
7.8 pH-meter.
7.9 Magnetic stirrer or shaking device.
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8 Procedure
8.1 Test material

The test material shall be of known mass and contain sufficient carbon to yield CO in a quantity

that can be adequately measured by the analytical system used. Calculate the TOC from the chemical

formula or determine it by a suitable analytical technique (e.g. elemental analysis or measurement in

accordance with ISO 8245) and calculate the ThCO . Use a concentration of a test material such that

the TOC content is at least 30 mg/l preferably 100 mg/l. The maximum amount of test material is

limited by the oxygen supply to the test system and the test medium used. When using higher amount

of test material, the optimized test medium (6.2.2) should be used and, in any case the test-material

concentration shall be such that the TOC does not exceed about 2 000 mg/l, i.e. a C:N ratio of about 40:1.

If higher concentrations are to be tested, increase the nitrogen amount in the test medium.

NOTE Lower amount of test materials, corresponding to a TOC content between 20 mg/l and 40 mg/l, have

been tested and found suitable.

The test material should preferably be used in powder form, but it may also be introduced as films,

pieces, fragments or shaped articles. The form and shape of the test material may influence its

biodegradation. Similar shapes should preferably be used if different kinds of plastic material are

to be compared. If the test material is used in the form of a powder, particles of known, narrow size

distribution should be used. A particle-size distribution with the maximum at 250 μm diameter is

recommended. Also, the size of the test equipment used may depend on the form of the test material.

It should be ascertained that no substantial mechanical aberrations occur due to the test conditions,

for example due to the type of stirring mechanism used. Processing of the test material (e.g. the use of

powder in the case of composites) should not influence significantly the degradation behaviour of the

material. Optionally, record the hydrogen, oxygen, nitrogen, phosphorus and sulfur contents and the

molecular mass of a polymeric test material, using for example liquid exclusion chromatography (see,

[7]

for example, ASTM D 3536–91 or any other applicable standard method). Preferably, plastic materials

without additives such as plasticizers should be tested. When the material does contain such additives,

information on their biodegradability will be needed to assess the biodegradability of the polymeric

material itself.
For details on how to h
...

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