SIST-TS CEN ISO/TS 29843-1:2015
(Main)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)
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)
This part of ISO/TS 29843 specifies an extended method for the extraction and determination of both hospholipid fatty acids (PLFA) and phospholipid ether lipids (PLEL) from soils. ISO/TS 29843-2 specifies a simple method for the extraction of only PLFA from soils.
Bodenbeschaffenheit - Bestimmung der Diversität von Bodenmikroorganismen - Teil 1: Verfahren mittels Phospholipidfettsäure(PLFA)-Analyse und Phospholipidetherlipid(PLEL)-Analyse (ISO/TS 29843-1:2010)
Dieser Teil von ISO/TS 29843 legt ein erweitertes Verfahren zur Extraktion und Bestimmung von Phospho¬lipid-fettsäuren (PLFA) und Phospholipidetherlipiden (PLEL) aus Böden fest.
ISO/TS 29843 2 beschreibt ein einfaches Verfahren zur Extraktion ausschließlich von PLFA aus Böden.
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)
L'ISO/TS 29843-1:2010 spécifie une méthode étendue permettant d'extraire et de doser les acides gras phospholipidiques (PLFA) et les lipides éther phospholipidiques (PLEL) du sol.
L'ISO/TS 29843-2 spécifie une méthode simple permettant d'extraire uniquement les PLFA du sol.
Kakovost tal - Določanje mikrobne raznolikosti tal - 1. del: Analiza fosfolipidnih maščobnih kislin (PLFA) in fosfolipidnih etrov (PLEL) (ISO/TS 29843-1:2010)
Ta del standarda ISO/TS 29843 določa razširjeno metodo za ekstrakcijo in določanje
fosfolipidnih maščobnih kislin in fosfolipidnih etrov iz tal. Standard ISO/TS 29843-2 določa preprosto metodo samo za ekstrakcijo fosfolipidnih maščobnih kislin iz tal.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN ISO/TS 29843-1:2015
01-januar-2015
.DNRYRVWWDO'RORþDQMHPLNUREQHUD]QROLNRVWLWDOGHO $QDOL]DIRVIROLSLGQLK
PDãþREQLKNLVOLQ 3/)$ LQIRVIROLSLGQLKHWURY 3/(/ ,6276
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
SIST-TS CEN ISO/TS 29843-1:2015 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TS CEN ISO/TS 29843-1:2015
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SIST-TS CEN ISO/TS 29843-1:2015
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,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, 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: 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.
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SIST-TS CEN ISO/TS 29843-1:2015
CEN ISO/TS 29843-1:2014 (E)
Contents
Page
Foreword .3
2
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SIST-TS CEN ISO/TS 29843-1:2015
CEN ISO/TS 29843-1:2014 (E)
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.
3
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SIST-TS CEN ISO/TS 29843-1:2015
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SIST-TS CEN ISO/TS 29843-1:2015
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
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SIST-TS CEN ISO/TS 29843-1:2015
ISO/TS 29843-1:2010(E)
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ii © ISO 2010 – All rights reserved
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SIST-TS CEN ISO/TS 29843-1:2015
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
2
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 2010 – All rights reserved iii
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SIST-TS CEN ISO/TS 29843-1:2015
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
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.
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
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
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
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SIST-TS CEN ISO/TS 29843-1:2015
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-linked isoprenoid side chains (PLEL: phospholipid ether lipid) allow for
taxonomic differentiation within complex microbial communities (References [5] and [7]). This approach is now
well established in soil ecology and serves as a phenotypic and thus complementary tool to genotypic
(molecular genetic) approaches for determining microbial diversity.
Different methodologies for determination of soil fatty acids are available. These methodologies present
different levels of complexity when applied and provide different levels of resolution in the description of soil
microbial communities.
The determination of total PLFA and PLEL provides a quantitative measure of the viable biomass of soil:
microorganisms of all three domains of the biosphere (bacteria, fungi and archaebacteria). Viable microbes
have an intact membrane, which contains phospholipids. Cellular enzymes hydrolyze and release the
phosphate group within minutes or hours following cell death (Reference [6]).
Apart from taxonomic descriptions, the PLFA technique enables the determination of physiological changes
within microbial consortia. For instance, the monoenic PLFA 16:1ω7c and 18:1 ω7c are increasingly converted
to the cyclopropyl fatty acids cy17:0 and cy19:0 in Gram-negative bacteria in response to environmental
stress (Reference [2]).
Besides the method described in this part of ISO/TS 29843, other methods for the determination of PLFA are
available (References [3] and [6]). With these methods, only bacterial and fungal PLFA can be estimated; the
determination of hydroxy-substituted fatty acids (PLOH), non-ester-linked (NEL) fatty acids and PLEL is not
possible.
© ISO 2010 – All rights reserved v
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SIST-TS CEN ISO/TS 29843-1:2015
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SIST-TS CEN ISO/TS 29843-1:2015
TECHNICAL SPECIFICATION ISO/TS 29843-1:2010(E)
Soil quality — Determination of soil microbial diversity —
Part 1:
Method by phospholipid fatty acid analysis (PLFA)
and phospholipid ether lipids (PLEL) analysis
1 Scope
This part of ISO/TS 29843 specifies an extended method for the extraction and determination of both
phospholipid fatty acids (PLFA) and phospholipid ether lipids (PLEL) from soils.
ISO/TS 29843-2 specifies a simple method for the extraction of only PLFA from soils.
2 Normative references
The following referenced documents are indispensable for the application 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 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil
under aerobic conditions for the assessment of microbiological processes, biomass and diversity in the
laboratory
3 Abbreviated terms
FAME fatty acid methyl ester(s)
(EL-)PLFA (ester-linked) phospholipid fatty acid(s)
PLEL phospholipid ether lipid(s)
SATFA saturated fatty acid(s)
MUFA mono-unsaturated fatty acid(s)
PUFA poly-unsaturated fatty acid(s)
PLOH hydroxy-substituted fatty acid(s)
NEL-PLFA non-ester-linked phospholipid fatty acid(s)
UNSFA unsubstituted fatty acid(s)
UNOH hydroxy-substituted fatty acid(s)
GC/MS gas chromatography/mass spectrometry
© ISO 2010 – All rights reserved 1
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