ISO/TS 5094:2023

TECHNICAL ISO/TS
SPECIFICATION 5094
First edition
2023-02
Nanotechnologies — Assessment of
peroxidase-like activity of metal and
metal oxide nanoparticles
Nanotechnologies — Evaluation de l'activité de type peroxidase des
nanoparticules métalliques et d’oxydes métalliques
Reference number
© ISO 2023
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ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms, definitions and abbreviated terms . 1
3.1 Terms and definitions . 1
3.2 Abbreviated terms . 2
4 Principle . 2
5 Physicochemical characterization of metal or metal oxide NPs . 3
6 Apparatus and reagents .4
6.1 Apparatus and appliances . 4
6.2 Reagents . 4
7 Solution preparation .5
7.1 General requirements . 5
7.2 TMB solution . 5
7.3 Buffer solution . 5
7.4 Nanoparticle dispersion solution . 5
7.5 Additional control solution . 5
8 Measurement procedure . 6
8.1 Measurement condition . 6
8.2 Measurement procedure . 6
8.3 Measurement of reagent blank absorption . 7
8.4 Positive control measurement . . 7
8.5 Additional control measurement . 7
9 Data analysis . 7
10 Measurement uncertainties . 8
11 Test report . 9
Annex A (informative) Measurement for the mass from the tested metal and metal oxide
nanoparticles .10
Annex B (informative) Example of calculation and uncertainty evaluation from the
peroxidase-like activity of iron oxide nanoparticles .11
Bibliography .17
iii
Foreword
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iv
Introduction
Enzymes are the biological catalysts that control biochemical reactions. The enzyme peroxidase is a
metalloenzyme with many isoforms. It catalyses the oxidation of various organic substrates by hydrogen
peroxide, which is used extensively in biochemistry applications. Metal and metal oxide nanoparticles
have a wide range of applications in biomedicine, environment protection, and some other fields, such
as magnetic separation, detection, anti-bacterial, degradation of contaminants, medical imaging and
tumour therapy. In recent years, an intrinsic peroxidase-like activity was observed in some metal and
metal oxide nanoparticles, which means that these metal and metal oxide nanoparticles can catalyse
the oxidation of substrates of natural peroxidase by hydrogen peroxide under mild reaction conditions
in comparable efficiency and kinetics. Iron oxide (Fe O ) nanoparticles are one representative
3 4
material, and cobalt oxide (Co O ) nanoparticles, copper oxide (CuO) nanoparticles, manganese oxide
3 4
(MnO ) nanoparticles, vanadium oxide (V O ) nanoparticles, gold (Au) nanoparticles and platinum (Pt)
2 2 5
nanoparticles have been reported to have the peroxidase-like activity as well. These findings extend
enzyme mimics from organic compounds to inorganic nanomaterials.
Certain metal and metal oxide nanoparticles can catalyse the transfer of electrons from H O to
2 2
colorimetric indicator under physiological condition. This phenomenon is like the colorimetric reaction
mediated by peroxidase and thus is called as peroxidase-like catalysis. Such catalytic property can
be used to produce colorimetric, chemiluminescent or electrochemical signals which have great
potential applications in biosensors, electrochemical sensors and immunoassays. The nanoparticles
with peroxidase-like activity may have anti-tumour, antibacterial or antioxidant functions in biological
system. In addition, the nanoparticles with such activity can have potential impacts on health, safety
and the environment. Therefore, it is important to assess the peroxidase-like activity of a nanoparticle
in practical applications.
The peroxidase-like activity of nanoparticles strongly depends on multiple factors including the
composition, size, surface chemistry and crystal structure of the nanoparticles, as well as the
measurement conditions. Therefore, it is important to establish a standard method for assessing the
peroxidase-like activity of metal and metal oxide nanoparticles.
This document provides a specification for the assessment of peroxidase-like activity of metal and
metal oxide nanoparticles. This protocol is useful to enterprises, research laboratories or institutions
and metrological organizations that are working on nanomaterials used in biomedical applications and
environment protection.
v
TECHNICAL SPECIFICATION ISO/TS 5094:2023(E)
Nanotechnologies — Assessment of peroxidase-like
activity of metal and metal oxide nanoparticles
1 Scope
This document specifies a method for assessing the peroxidase-like activity of metal and metal oxide
nanoparticles by spectrophotometry. This document can serve as a reference for the measurements of
peroxidase-like activities in other types of nanoparticles.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements 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 18153:2003, In vitro diagnostic medical devices — Measurement of quantities in biological samples —
Metrological traceability of values for catalytic concentration of enzymes assigned calibrators and control
materials
ISO/TS 80004-2, Nanotechnologies — Vocabulary — Part 2: Nano-objects
3 Terms, definitions and abbreviated terms
For the purposes of this document, the terms and definitions given in ISO/TS 80004-2 and the following
apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1 Terms and definitions
3.1.1
nanoparticle
nano-object with all external dimensions in the nanoscale where the lengths of the longest and the
shortest axes of the nano-object do not differ significantly
[SOURCE: ISO/TS 80004-2:2015, 4.4, modified — Note 1 to entry has been removed.]
3.1.2
catalytic activity
property of a component corresponding to the catalysed substance rate of conversion of a specified
chemical reaction, in a specified measurement system
Note 1 to entry: In this document, the “component” is one kind of metal or metal oxide nanoparticles.
Note 2 to entry: In this document, the catalytic activity is the peroxidase-like activity of metal and metal oxide
nanoparticles.
−1
Note 3 to entry: The coherent derived SI unit is “katal” (kat), equal to “mole per second” (mol·s ).
[SOURCE: ISO 18153:2003, 3.2, modified — Notes 1, 2 and 3 to entry have been added.]
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