ISO TS 18827-2017 Nanotechnologies - Electron spin resonance (ESR) as a method for measuring reactive oxygen species (ROS) generated by metal oxide nanomaterial.pdf
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1、 ISO 2017 Nanotechnologies Electron spin resonance (ESR) as a method for measuring reactive oxygen species (ROS) generated by metal oxide nanomaterials Nanotechnologies Rsonance paramagntique lectronique (RPE) pour la mesure des espces ractives de loxygne (ROS) gnres par des nanomatriaux sous forme
2、doxyde mtallique TECHNICAL SPECIFICATION ISO/TS 18827 Reference number ISO/TS 18827:2017(E) First edition 2017-06 ISO/TS 18827:2017(E)ii ISO 2017 All rights reserved COPYRIGHT PROTECTED DOCUMENT ISO 2017, Published in Switzerland All rights reserved. Unless otherwise specified, no part of this publi
3、cation 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 ISOs member body in the count
4、ry of the requester. ISO copyright office Ch. de Blandonnet 8 CP 401 CH-1214 Vernier, Geneva, Switzerland Tel. +41 22 749 01 11 Fax +41 22 749 09 47 copyrightiso.org www.iso.org ISO/TS 18827:2017(E)Foreword v Introduction vi 1 Scope . 1 2 Normative references 1 3 T erms, definitions and abbr e viati
5、ons 1 3.1 Terms and definitions . 1 3.2 Abbreviations . 2 4 Principle 2 4.1 General . 2 4.2 Spin trapping method . 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 Hypoxanthinexanthine oxidase syst
6、em 15. 3 4.3.3 Rose bengal photosensitization 16174 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 FeSO 4solution 5 7.2.2 H 2 O 2solution 5 7.3 Preparation of
7、 solution for generating the superoxide anion radical . 5 7.3.1 Phosphate buffer 5 7.3.2 Hypoxanthine solution . 5 7.3.3 Xanthine oxidase solution 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
8、7.5.3 BMPO stock solution. 6 7.5.4 TPC stock solution 6 7.6 Reaction of test sample and spin trapping agent 6 7.6.1 General 6 7.6.2 DMPO reaction . 6 7.6.3 BMPO reaction . 7 7.6.4 TPC reaction . 7 7.7 Reaction of positive control and spin trapping agent 7 7.7.1 DMPO radical adduct form (DMPO/OH) . 7
9、 7.7.2 BMPO radical adduct form (BMPO/OOH) 7 7.7.3 TPC radical adduct form (TPC/ 1 O 2 ) 7 7.8 Preparation of the standard sample for spin calculation . 7 8 Interferences 8 8.1 Sampling . 8 8.2 Sampling time 8 9 Procedure. 8 9.1 General . 8 9.2 Injection of sample 9 ISO 2017 All rights reserved iii
10、Contents Page ISO/TS 18827:2017(E)9.3 ESR measurement .10 10 Examples of results .16 10.1 DMPO/OH .16 10.2 BMPO/OOH .16 10.3 TPC/ 1 O 216 10.4 TEMPOL .17 Bibliography .18 iv ISO 2017 All rights reserved ISO/TS 18827:2017(E) Foreword ISO (the International Organization for Standardization) is a world
11、wide 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 t
12、hat 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 do
13、cument 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 Direct
14、ives, 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 deve
15、lopment 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 on the voluntary
16、nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following URL: w w w . i s o .org/ iso/ foreword .htm
17、l. This document was prepared by Technical Committee ISO/TC 229, Nanotechnologies. ISO 2017 All rights reserved v ISO/TS 18827:2017(E) Introduction Recently, the use of metal or metal oxide-based nanomaterials has dramatically increased in biomedical and industrial applications. However, the scienti
18、fic basis for the cytotoxicity and genotoxicity of most manufactured nanomaterials are not fully understood. An important mechanism of nanotoxicity is the generation of reactive oxygen species (ROS). The study on the hazardous effects of metal oxide nanomaterials is still in its initial stage. The a
19、bility to generate ROS is one main source of toxicity of metal oxide nanomaterials. Overproduction of ROS can induce oxidative stress, resulting in cells failing to maintain normal physiological redox-regulated functions. This in turn may lead to DNA damage, unregulated cell signalling, change in ce
20、ll motility, cytotoxicity, apoptosis and cancer initiation. There are critical determinants that can affect the generation of ROS. The critical determinants include size, shape, particle surface, surface positive charges, surface-containing groups, particle dissolution, metal ion release from nanome
21、tals and nanometal oxides, UV light activation, aggregation, mode of interaction with cells, inflammation and pH of the medium 1 . Thus, to detect and quantify ROS formation on the surface of metal oxide nanomaterials, this document suggests the electron-spin-resonance (ESR) method. Amongst ROS, the
22、 most biologically relevant and widely studied are hydroxyl radical (OH), superoxide anion radical (O 2 - ), singlet oxygen ( 1 O 2 ) and hydrogen peroxide (H 2 O 2 ). However, direct detection of some free radicals (e.g. superoxide anion and hydroxyl radical) is very difficult or impossible 2in sol
23、ution at room temperature. ESR spin trapping is a valuable tool in the study of transient free radicals 3 . Spin trapping is a technique, developed in the late 1960s, where a nitrone or nitroso compound (a spin trap) reacts with a target free radical to form a stable and distinguishable free radical
24、 (spin adducts) to be detected by ESR spectroscopy. Spin adducts can be observed directly by ESR spectroscopy. The ESR spectra of these spin adducts are unique and provide a fingerprint for the presence of ROS. This document specifies methods of detection by ESR of 5,5-dimethyl-1-pyrroline-N-oxide (
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