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)

Kakovost tal - Določevanje mikrobne raznolikosti tal - 2. del: Analiza fosfolipidnih maščobnih kislin (PLFA) z enostavno ekstrakcijsko metodo PLFA (ISO/TS 29843-2:2021)

General Information

Status
Published
Public Enquiry End Date
02-May-2021
Publication Date
11-Oct-2021
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
18-Aug-2021
Due Date
23-Oct-2021
Completion Date
12-Oct-2021

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SLOVENSKI STANDARD
SIST-TS CEN ISO/TS 29843-2:2021
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
SIST-TS CEN ISO/TS 29843-2:2021 en,fr,de

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

---------------------- Page: 1 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
---------------------- Page: 2 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
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.

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. CEN ISO/TS 29843-2:2021 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
CEN ISO/TS 29843-2:2021 (E)
Contents Page

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

---------------------- Page: 4 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
CEN ISO/TS 29843-2:2021 (E)
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.
---------------------- Page: 5 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
---------------------- Page: 6 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
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
---------------------- Page: 7 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
ISO/TS 29843-2:2021(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
---------------------- Page: 8 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
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 2021 – All rights reserved iii
---------------------- Page: 9 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
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
---------------------- Page: 10 ----------------------
SIST-TS CEN ISO/TS 29843-2:2021
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 . Furthermore, combined with the use of isotope ( C or C) labelled substrates, the

lipid methods can also be used to identify the metabolically active part of the microbial community.

This approach is well established in soil ecology and serves as a phenotypic, and thus complementary,

tool to genotyping approaches for determining microbial diversity. Apart from taxonomic descriptions,

the PLFA technique enables the determination of physiological changes within microbial consortia. For

instance, the monoenic PLFA 16:1ω7c and 18:1ω7c are increasingly converted to the cyclopropyl fatty

[7]

acids cy17:0 and cy19:0 in Gram-negative bacteria in response to environmental stress .

Different methodologies are available for the determination of soil fatty acids. These methodologies

present different levels of complexity when applied and provide different levels of resolution in the

description of soil microbial communities. ISO/TS 29843-1 deals with the generally called “extended

PLFA extraction method” while this document deals with the generally called “simple PLFA extraction

[8],[9]
method” .

This document is accessible to most research and analytical laboratories involved in soil sciences. This

methodology can be used for a wide range of soils. It provides a broad diversity measurement of a soil

microbial community at the phenotypic level. It can be applied to biomass estimation and can be used to

differentiate microbial communities among different soil samples (with the aid of an adapted statistical

method). This method is especially adapted for detecting rapid changes in the soil microbial community

structure. It can also be used to give a rough description of microbial groups present in soil samples

[6]

(e.g. Gram-positive bacteria, actinomycetes, fungi ). Table 1 (adapted from Table 1 in Reference [8])

...

SLOVENSKI STANDARD
kSIST-TS FprCEN ISO/TS 29843-2:2021
01-april-2021

Kakovost tal - Določanje mikrobne raznolikosti tal - 2. del: Analiza fosfolipidnih

maščobnih kislin (PLFA) z enostavno ekstrakcijsko metodo PLFA (ISO/PRF 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/PRF TS 29843-

2:2021)

Bodenbeschaffenheit - Bestimmung der Diversität von Bodenmikroorganismen - Teil 2:

Verfahren mittels Phospholipidfettsäure(PLFA)-Analyse unter Verwendung des einfachen

PLFA-Extraktionsverfahrens (ISO/PRF 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/PRF TS 29843-2:2021)
Ta slovenski standard je istoveten z: FprCEN ISO/TS 29843-2
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
kSIST-TS FprCEN ISO/TS 29843-2:2021 en,fr,de

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

---------------------- Page: 1 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
---------------------- Page: 2 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
TECHNICAL ISO/TS
SPECIFICATION 29843-2
Second edition
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
PROOF/ÉPREUVE
Reference number
ISO/TS 29843-2:2021(E)
ISO 2021
---------------------- Page: 3 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
ISO/TS 29843-2:2021(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 PROOF/ÉPREUVE © ISO 2021 – All rights reserved
---------------------- Page: 4 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
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 2021 – All rights reserved PROOF/ÉPREUVE iii
---------------------- Page: 5 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
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 PROOF/ÉPREUVE © ISO 2021 – All rights reserved
---------------------- Page: 6 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
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 . Furthermore, combined with the use of isotope ( C or C) labelled substrates, the

lipid methods can also be used to identify the metabolically active part of the microbial community.

This approach is well established in soil ecology and serves as a phenotypic, and thus complementary,

tool to genotyping approaches for determining microbial diversity. Apart from taxonomic descriptions,

the PLFA technique enables the determination of physiological changes within microbial consortia. For

instance, the monoenic PLFA 16:1ω7c and 18:1ω7c are increasingly converted to the cyclopropyl fatty

[7]

acids cy17:0 and cy19:0 in Gram-negative bacteria in response to environmental stress .

Different methodologies are available for the determination of soil fatty acids. These methodologies

present different levels of complexity when applied and provide different levels of resolution in the

description of soil microbial communities. ISO/TS 29843-1 deals with the generally called “extended

PLFA extraction method” while this document deals with the generally called “simple PLFA extraction

[8],[9]
method” .

This document is accessible to most research and analytical laboratories involved in soil sciences. This

methodology can be used for a wide range of soils. It provides a broad diversity measurement of a soil

microbial community at the phenotypic level. It can be applied to biomass estimation and can be used to

differentiate microbial communities among different soil samples (with the aid of an adapted statistical

method). This method is especially adapted for detecting rapid changes in the soil microbial community

structure. It can also be used to give a rough description of microbial groups present in soil samples

[6]

(e.g. Gram-positive bacteria, actinomycetes, fungi ). Table 1 (adapted from Table 1 in Reference [8]),

presents a comparison of the sensitivity of the “extended PLFA” versus “simple PLFA” techniques.

Table 1 — Comparison of the sensitivity of the “simple” and “extended” PLFA techniques

in characterizing shifts in the composition of microbial communities
Property PLFA (simple) PLFA (extended)
Ability to differentiate between two communities Yes Yes
(with the aid of multivariate statistical methods)
Applicability for biomass estimation Yes Yes
Ability to register all single components of an entire No Yes
community structure (“fingerprint”)
Ability to register FAs other than EL-FAs No Yes
Estimation of number of FAs in soil samples <50 200 to 400
Capacity to determine the linkage of the FAs to lipids Yes, EL Yes, EL, NEL
in the molecule
Capacity to detect defined FAs in lower concentrations No Yes
in the soil extract
Capacity to detect unusual FAs in the soil extract No Yes
Number of available signatures of FAs for defined Few High numbers
organisms
Relationships of FAs widespread in the profile High Natura
© ISO 2021 – All rights reserved PROOF/ÉPREUVE v
---------------------- Page: 7 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
ISO/TS 29843-2:2021(E)
Table 1 (continued)
Property PLFA (simple) PLFA (extended)
Ability to identify the organisms causing the shift in No Basically yes
microbial community
FA fatty acid
EL ester-linked
NEL non-ester–linked

This method has been derived from the one first proposed in Reference [10]. This revised method

[11]

has been widely used and has also been discussed and compared to the extended PLFA extraction

[8],[9]
method in peer-reviewed articles .
vi PROOF/ÉPREUVE © ISO 2021 – All rights reserved
---------------------- Page: 8 ----------------------
kSIST-TS FprCEN ISO/TS 29843-2:2021
TECHNICAL SPECIFICATION ISO/TS 29843-2:2021(E)
Soil quality — Determination of soil microbial diversity —
Part 2:
Method by phospholipid fatty acid analysis (PLFA) using
the simple PLFA extraction method
1 Scope

This document specifies a simple method for the extraction of only phospholipid fatty acids (PLFA)

from soils.
2 Normative references
The following documents are referred to in the
...

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