CEN ISO/TS 29843-1:2014

SLOVENSKI STANDARD
01-januar-2015
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Soil quality - Determination of soil microbial diversity - Part 1: Method by phospholipid
fatty acid analysis (PLFA) and phospholipid ether lipids (PLEL) analysis (ISO/TS 29843-
1:2010)
Qualité du sol - Détermination de la diversité microbienne du sol - Partie 1: Méthode par
analyse des acides gras phospholipidiques (PLFA) et par analyse des lipides éther
phospholipidiques (PLEL) (ISO/TS 29843-1:2010)
Ta slovenski standard je istoveten z: CEN ISO/TS 29843-1:2014
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.

TECHNICAL SPECIFICATION
CEN ISO/TS 29843-1
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
October 2014
ICS 13.080.30
English Version
Soil quality - Determination of soil microbial diversity - Part 1:
Method by phospholipid fatty acid analysis (PLFA) and
phospholipid ether lipids (PLEL) analysis (ISO/TS 29843-1:2010)
Qualité du sol - Détermination de la diversité microbienne Bodenbeschaffenheit - Bestimmung der Diversität von
du sol - Partie 1: Méthode par analyse des acides gras Bodenmikroorganismen - Teil 1: Verfahren mittels
phospholipidiques (PLFA) et par analyse des lipides éther Phospholipidfettsäure(PLFA)-Analyse und
phospholipidiques (PLEL) (ISO/TS 29843-1:2010) Phospholipidetherlipid(PLEL)-Analyse (ISO/TS 29843-
1:2010)
This Technical Specification (CEN/TS) was approved by CEN on 11 August 2014 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.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
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Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 29843-1:2014 E
worldwide for CEN national Members.

Contents
Page
Foreword .3

Foreword
The text of ISO/TS 29843-1:2010 has been prepared by Technical Committee ISO/TC 190 "Soil quality" of the
International Organization for Standardization (ISO) and has been taken over as CEN ISO/TS 29843-1:2014
by Technical Committee CEN/TC 345 “Characterization of soils” 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 [and/or CENELEC] 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 announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO/TS 29843-1:2010 has been approved by CEN as CEN ISO/TS 29843-1:2014 without any
modification.
TECHNICAL ISO/TS
SPECIFICATION 29843-1
First edition
2010-10-01
Soil quality — Determination of soil
microbial diversity —
Part 1:
Method by phospholipid fatty acid
analysis (PLFA) and phospholipid ether
lipids (PLEL) analysis
Qualité du sol — Détermination de la diversité microbienne du sol —
Partie 1: Méthode par analyse des acides gras phospholipidiques
(PLFA) et par analyse des lipides éther phospholipidiques (PLEL)

Reference number
ISO/TS 29843-1:2010(E)
©
ISO 2010
ISO/TS 29843-1:2010(E)
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ii © ISO 2010 – All rights reserved

ISO/TS 29843-1:2010(E)
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Abbreviated terms .1
4 Principle.2
5 Reagents and materials .3
5.1 Soil .3
5.2 Reagents.3
5.3 Buffers and standards .4
5.4 Apparatus.4
6 Procedures.5
6.1 Lipid extraction (Bligh-Dyer-extraction).5
6.2 Separation of lipids by sI-column.5
6.3 PLFA analysis .5
6.3.1 Mild alkaline hydrolysis .5
6.3.2 NH column: Separation of FAME from OH-substituted FAME (= PLOH) and
unsaponifiable lipids.5
6.3.3 SCX column: Separation of unsubstituted ester-linked PLFA (EL-PLFA).6
6.3.4 Acidic methylation of unsaponifiable lipids and separation into UNOH and UNSFA .6
6.3.5 TMSI derivatization of PLOH and UNOH (see 5.2.22).6
6.3.6 DMDS derivatization of MUFA (see 5.2.8).6
6.4 PLEL analysis .7
6.4.1 General .7
6.4.2 Acidic methylation.7
6.4.3 Cleavage of etherbonds with hydroiodic acid (HI).7
6.4.4 Reductive dehalogenization with zinc.7
6.5 Measurement of PLFA/PLEL fractions .7
7 Identification and calculation.8
Bibliography.9

ISO/TS 29843-1:2010(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
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International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
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International Standard requires approval by at least 75 % of the member bodies casting a vote.
In other circumstances, particularly when there is an urgent market requirement for such documents, a
technical committee may decide to publish other types of document:
⎯ an ISO Publicly Available Specification (ISO/PAS) represents an agreement between technical experts in
an ISO working group and is accepted for publication if it is approved by more than 50 % of the members
of the parent committee casting a vote;
⎯ an ISO Technical Specification (ISO/TS) represents an agreement between the members of a technical
committee and is accepted for publication if it is approved by 2/3 of the members of the committee casting
a vote.
An ISO/PAS or ISO/TS is reviewed after three years in order to decide whether it will be confirmed for a
further three years, revised to become an International Standard, or withdrawn. If the ISO/PAS or ISO/TS is
confirmed, it is reviewed again after a further three years, at which time it must either be transformed into an
International Standard or be withdrawn.
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.
ISO/TS 29843-1 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological methods.
ISO/TS 29843 consists of the following parts, under the general title Soil quality — Determination of soil
microbial diversity:
⎯ Part 1: Method by phospholipid fatty acid analysis (PLFA) and phospholipid ether lipids (PLEL) analysis
⎯ Part 2: Method by phospholipid fatty acid analysis (PLFA) using the “simple PLFA extraction method”
iv © ISO 2010 – All rights reserved

ISO/TS 29843-1:2010(E)
Introduction
Phospholipids are essential components of membranes of all living cells, and their fatty acid (PLFA:
phospholipid fatty acids) or ether-linke
...