ISO/TS TECHNICAL SPECIFICATION 18827 First edition 2017-06 Nanotechnologies Electron spin resonance (EsR) as a method for measuring reactive oxygen species (ROS) generated by metal oxide nanomaterials Nanotechnologies - Résonance paramagnétique électronique (RPE) pour la mesure des especes réactives de I'oxygene (ROs) générées par des nanomateriaux sous forme d'oxyde métallique Reference number IS0/TS 18827:2017(E) International Organization for Standardization @ IS0 2017 =ZHEJIANG INST OF STANDARDIZATION C1 5956617 HS unde ted without license from IHS IS0/TS 18827:2017(E) COPYRIGHT PROTECTED DOCUMENT IS0 2017, Published in Switzerland All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form 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 copyrightoffice Ch. de Blandonnet 8 . CP 401 CH-1214 Vernier, Geneva, Switzerland Tel. +41 22 749 01 11 Fax +41 22 749 09 47 [email protected] www.iso.org Internatinir DrganizationforStandardization Licensee-ZHEJIANG INST OF STANDARDIZzRoISQ7 - All rights reserved No reproduction or networking permited without license from IHS IS0/TS 18827:2017(E) Contents Page Foreword ..V Introduction.. ..vi 1 Scope. 2 Normative references 3 Terms, definitions and abbreviations 3.1 Terms and definitions .1 3.2 Abbreviations... 4 Principle. .2 4.1 General .2 Spin trapping method .2 4.2 4.2.1 General. 2 4.2.2 DMPO ..2 4.2.3 BMPO .3 4.2.4 TPC .3 4.3 Positive control for generating free radicals .3 4.3.1 Fenton reaction[14] 3 4.3.2 Hypoxanthine-xanthine oxidase system[15] 3 4.3.3 Rose bengal photosensitization[16][1Z]. .4 5 Reagents. .4 6 Apparatus .4 7 Sampling. ..5 7.1 Preparation of test sample (metal oxide nanomaterial suspension) .5 7.2 Preparation of solution for generating the hydroxyl radical 5 7.2.1 FeSO4 solution. .5 7.2.2 H202 solution. .5 7.3 Preparation of solution for generating the superoxide anion radical 7.3.1 Phosphate buffer. .5 7.3.2 Hypoxanthine solution 5 7.3.3 Xanthineoxidasesolution 5 7.4 Preparation of solution for generating the singlet oxygen. .5 7.5 Preparation of spin trapping agent .6 7.5.1 General. .6 7.5.2 DMPO stock solution ..6 7.5.3 BMPO stock solution. .6 7.5.4 TPC stock solution. 7.6 Reaction of test sample and spin trapping agent .6 7.6.1 General. 7.6.2 DMPO reaction. 6 7.6.3 BMPO reaction 7.6.4 TPCreaction. 7 7.7 Reaction of positive control and spin trapping agent 7.7.1 DMPO radical adduct form (DMPO/OH) 7 7.7.2 BMPO radical adduct form (BMPO/OOH) .7 7.7.3 TPC radical adduct form (TPC/102) 7.8 Preparation of the standard sample for spin calculation .7 8 Interferences .8 8.1 Sampling. 8.2 Sampling time ..8 9 Procedure. 9.1 General. 9.2 Injection of sample .9 ii icensee=ZHEJIANG INST OF STANDARDIZATION C1 5956617 vided by IHS under license with ISO without license from IHS