CEN ISO/TS 29843-2:2021

SLOVENSKI STANDARD
01-november-2021
Nadomešča:
SIST-TS CEN ISO/TS 29843-2:2014
Kakovost tal - Določevanje mikrobne raznolikosti tal - 2. del: Analiza fosfolipidnih
maščobnih kislin (PLFA) z enostavno ekstrakcijsko metodo PLFA (ISO/TS 29843-
2:2021)
Soil quality - Determination of soil microbial diversity - Part 2: Method by phospholipid
fatty acid analysis (PLFA) using the simple PLFA extraction method (ISO/TS 29843-
2:2021)
Bodenbeschaffenheit - Bestimmung der Diversität von Bodenmikroorganismen - Teil 2:
Verfahren mittels Phospholipidfettsäure(PLFA)-Analyse unter Verwendung des einfachen
PLFA-Extraktionsverfahrens (ISO/TS 29843-2:2021)
Qualité du sol - Détermination de la diversité microbienne du sol - Partie 2: Méthode par
analyse des acides gras phospholipidiques (PLFA) en utilisant la méthode simple
d'extraction des PLFA (ISO/TS 29843-2:2021)
Ta slovenski standard je istoveten z: CEN ISO/TS 29843-2:2021
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.

CEN ISO/TS 29843-2
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
July 2021
TECHNISCHE SPEZIFIKATION
ICS 13.080.30 Supersedes CEN ISO/TS 29843-2:2014
English Version
Soil quality - Determination of soil microbial diversity -
Part 2: Method by phospholipid fatty acid analysis (PLFA)
using the simple PLFA extraction method (ISO/TS 29843-
2:2021)
Qualité du sol - Détermination de la diversité Bodenbeschaffenheit - Bestimmung der Diversität von
microbienne du sol - Partie 2: Méthode par analyse des Bodenmikroorganismen - Teil 2: Verfahren mittels
acides gras phospholipidiques (PLFA) en utilisant la Phospholipidfettsäure(PLFA)-Analyse unter
méthode simple d'extraction des PLFA (ISO/TS 29843- Verwendung des einfachen PLFA-
2:2021) Extraktionsverfahrens (ISO/TS 29843-2:2021)
This Technical Specification (CEN/TS) was approved by CEN on 28 June 2021 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

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United Kingdom.
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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. CEN ISO/TS 29843-2:2021 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (CEN ISO/TS 29843-2:2021) has been prepared by Technical Committee ISO/TC 190
"Soil quality" in collaboration with Technical Committee CEN/TC 444 “Environmental characterization
of solid matrices” the secretariat of which is held by NEN.
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 ISO/TS 29843-2:2014.
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 announce this Technical Specification: 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/TS 29843-2:2021 has been approved by CEN as CEN ISO/TS 29843-2:2021 without any
modification.
TECHNICAL ISO/TS
SPECIFICATION 29843-2
Second edition
2021-07
Soil quality — Determination of soil
microbial diversity —
Part 2:
Method by phospholipid fatty acid
analysis (PLFA) using the simple PLFA
extraction method
Qualité du sol — Détermination de la diversité microbienne du sol —
Partie 2: Méthode par analyse des acides gras phospholipidiques
(PLFA) en utilisant la méthode simple d'extraction des PLFA
Reference number
ISO/TS 29843-2:2021(E)
©
ISO 2021
ISO/TS 29843-2:2021(E)
© ISO 2021
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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ii © ISO 2021 – All rights reserved

ISO/TS 29843-2:2021(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Symbols and abbreviated terms (except chemical products and reagents) .1
5 Principle . 2
6 Test materials . 2
6.1 Soil . 2
6.2 Reagents. 2
6.3 Apparatus . 4
7 Procedures . 5
7.1 Lipid extraction (Bligh-Dyer extraction) . 5
7.2 Separation of lipids by SI column . 5
7.3 Derivatization — Transmethylation — Clean-up . 6
7.4 PLFA analysis . 6
Bibliography . 7
ISO/TS 29843-2:2021(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 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 190, Soil quality, Subcommittee SC 4,
Biological characterization, in collaboration with the European Committee for Standardization (CEN)
Technical Committee CEN/TC 444, Environmental characterization of solid matrices, in accordance with
the Agreement on technical cooperation between ISO and CEN (Vienna Agreement).
This second edition cancels and replaces the first edition (ISO/TS 29843-2:2011), which has been
technically revised.
The main changes compared to the previous edition are as follows:
— addition of specification for qualitative and quantitative analysis of PLFAs;
— use of BAME (qualitative) or FAME (quantitative) standards;
— use of GC-MS apparatus;
— precisions in 7.2 and 7.3;
— possibility to use commercial cartridges in addition to, or replacement of, home-made cartridges;
— update of bibliographic references.
A list of all the parts in the ISO/TS 29843 series can be found on the ISO website.
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

ISO/TS 29843-2:2021(E)
Introduction
Phospholipids are essential components of membranes of all living cells. Extracted from soil samples
in fatty acid form (phospholipid fatty acids, PLFA) or ether-linked isoprenoid side chains (phospholipid
ether lipid, PLEL), they provide quantitative and qualitative insights into the soil's viable/active
microbial biomass. Cellular enzymes hydrolyse and release the phosphate group within minutes to
[1]
hours following cell death . The determination of total PLFA and PLEL provides a quantitative measure
of the viable biomass of soil, i.e. microorganisms of all three primary domains of the biosphere (bacteria,
archaea and microeukaryota). PLFA and PLEL can also allow for rough taxonomic differentiation within
[2],[3]
complex microbial communities . Each microbial species contains several fatty acids, with a total
[4]
composition in PLFA subject to the environmental conditions . The approach is performed to evaluate
[5]
biomass and shifts in microbial community composition , in what regards dominance of main groups
[6] 13 14
of organisms . Furthermor
...