ISO/TS TECHNICAL 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: Methode par analyse des acides gras phospholipidiques (PLFA) et par analyse des lipides éther phospholipidiques (PLEL) Reference number ISO/TS 29843-1:2010(E) O @ ISO 2010 HS under I without license from IHS Not for Resale ISO/TS 29843-1:2010(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by IsO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below COPYRIGHTPROTECTEDDOCUMENT @ISO2010 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, isO's member body in the country of the requester. ISO copyright office Case postale 56. CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail [email protected] Web www.iso.org Published in Switzerland @ ISO 2010 - All rights reserved I without license from IHS Not for Resale ISO/TS 29843-1:2010(E) Contents Page Foreword .iv Introduction 1 Scope 2 Normative references. 3 Abbreviated terms 4 Principle... 5 Reagents and materials 5.1 Soil.. 5.2 Reagents. 5.3 Buffers and standards 5.4 Apparatus.. 6 Procedures. 6.1 Lipid extraction (Bligh-Dyer-extraction). 6.2 Separation of lipids by sl-column.... 6.3 PLFA analysis .... 6.3.1 Mild alkaline hydrolysis .. 6.3.2 NH2 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).... 6.3.4 Acidic methylation of unsaponifiable lipids and separation into UNOH and UNSFA 6.3.5 TMSI derivatization of PLOH and UNOH (see 5.2.22)... 6.3.6 DMDS derivatization of MUFA (see 5.2.8).. 6.4 PLEL analysis ... 6.4.1 General 6.4.2 Acidic methylation... 6.4.3 Cleavage of etherbonds with hydroiodic acid (Hl)..... 6.4.4 Reductive dehalogenization with zinc.. 6.5 Measurement of PLFA/PLEL fractions.... 7 Identification and calculation Bibliography. CopyrightInternatinal Oorganizain forStandardizalinghts reserved ii ted without license from IHS Not for Resale