SIST ISO 21338:2011
Water quality - Kinetic determination of the inhibitory effects of sediment, other solids and coloured samples on the light emission of Vibrio fischeri (kinetic luminescent bacteria test)
Water quality - Kinetic determination of the inhibitory effects of sediment, other solids and coloured samples on the light emission of Vibrio fischeri (kinetic luminescent bacteria test)
ISO 21338:2010 specifies the kinetic direct‑contact method for determining the inhibitory effect of suspensions of sediment and other solid samples, and also for problematic turbid or coloured aqueous samples on the light emission of the marine bacterium Vibrio fischeri (NRRL B‑11177).
This method is applicable to: a) sediment samples and water suspensions of sediments (fresh water, brackish, and seawater sediments); b) effluents (especially turbid and coloured); c) aqueous extracts (e.g. leachates, eluates, elutriates) of soil, solid waste, and other solid material (especially turbid and coloured).
Qualité de l'eau - Détermination cinétique des effets inhibiteurs des échantillons de sédiment, autres solides et des échantillons colorés sur la luminescence de Vibrio fischeri (essai cinétique de bactéries luminescentes)
Kakovost vode - Določevanje kinetike zaviralnih učinkov sedimenta, drugih trdnih in obarvanih vzorcev na emitirano svetlobo Vibrio fischeri (kinetic luminescent bacteria test)
Ta mednarodni standard opredeljuje kinetično metodo z neposrednim stikom za določevanje zaviralnih učinkov suspenzij sedimenta in drugih trdnih vzorcev ter tudi za problematične motne ali obarvane vodne vzorce na emitirano svetlobo morske bakterije Vibrio fischeri (NRRL B-11177).
Ta metoda se uporablja za:
a) sedimentne vzorce in vodne suspenzije sedimentov (sedimente sladke vode, nepitne vode in morske vode);
b) odplake (zlasti motne in obarvane);
c) vodne ekstrakte (npr. izlužki, eluati in elutriati), zemljine, trdne odpadke in druge trdne materiale(zlasti motne in obarvane).
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2011
.DNRYRVWYRGH'RORþHYDQMHNLQHWLNH]DYLUDOQLKXþLQNRYVHGLPHQWDGUXJLKWUGQLK
LQREDUYDQLKY]RUFHYQDHPLWLUDQRVYHWORER9LEULRILVFKHUL NLQHWLFOXPLQHVFHQW
EDFWHULDWHVW
Water quality - Kinetic determination of the inhibitory effects of sediment, other solids and
coloured samples on the light emission of Vibrio fischeri (kinetic luminescent bacteria
test)
Qualité de l'eau - Détermination cinétique des effets inhibiteurs des échantillons de
sédiment, autres solides et des échantillons colorés sur la luminescence de Vibrio
fischeri (essai cinétique de bactéries luminescentes)
Ta slovenski standard je istoveten z: ISO 21338:2010
ICS:
13.060.60 Preiskava fizikalnih lastnosti Examination of physical
vode properties of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
INTERNATIONAL ISO
STANDARD 21338
First edition
2010-07-15
Water quality — Kinetic determination of
the inhibitory effects of sediment, other
solids and coloured samples on the light
emission of Vibrio fischeri (kinetic
luminescent bacteria test)
Qualité de l'eau — Détermination cinétique des effets inhibiteurs des
échantillons de sédiment, autres solides et des échantillons colorés sur
la luminescence de Vibrio fischeri (essai cinétique de bactéries
luminescentes)
Reference number
©
ISO 2010
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
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
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.
© ISO 2010
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
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 copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2010 – All rights reserved
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Terms and definitions .2
4 Principle.2
5 Interferences .3
6 Reagents and materials .4
7 Apparatus.5
8 Sampling and sample pre-treatment .5
9 Procedure.6
10 Evaluation.7
11 Expression of results.9
12 Criteria of validity .11
13 Test report.11
Annex A (informative) Precision data .13
Annex B (informative) Typical kinetic curves from different samples .17
Annex C (informative) Dilution series .18
Bibliography.20
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.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
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.
ISO 21338 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
iv © ISO 2010 – All rights reserved
Introduction
The method specified in this International Standard is a kinetic modification of the luminescent bacteria test
specified in ISO 11348. The kinetic method overcomes the problems arising from intense colour and turbidity
in the samples. There is no need for sedimentation or centrifugation of turbid samples, or for the correction of
colour as described in ISO 11348.
This kinetic method uses luminometers capable of dispensing luminescent bacteria to the samples and
measuring the luminescent intensity over a period of time. In the method, the bacterial suspension is
dispensed and mixed with the sample in the measurement chamber of the luminometer. Several suitable
instruments are commercially available, but only a few of them are capable of cooling the measurement
chamber to (15 ± 1) °C as specified in ISO 11348. However, if the bacterial suspension and test samples are
kept at (15 ± 1) °C in a thermo-block before the measurement and during the whole incubation, the actual
temperature during the contact time is (15 ± 1) °C.
The measurements specified in this International Standard can be carried out using freshly prepared bacteria,
as well as freeze- or liquid-dried bacterial preparations. The various bacterial preparations can deliver different
results, especially in the presence of heavy metals (see ISO 11348). The laboratories responsible for the
results have the opportunity to select the most suitable bacterial preparation based on expert judgement and
information about the samples to be tested.
INTERNATIONAL STANDARD ISO 21338:2010(E)
Water quality — Kinetic determination of the inhibitory effects
of sediment, other solids and coloured samples on the light
emission of Vibrio fischeri (kinetic luminescent bacteria test)
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This International Standard does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this International
Standard be carried out by suitably trained staff.
1 Scope
This International Standard specifies the kinetic direct-contact method for determining the inhibitory effect of
suspensions of sediment and other solid samples, and also for problematic turbid or coloured aqueous
samples on the light emission of the marine bacterium Vibrio fischeri (NRRL B-11177).
This method is applicable to:
a) sediment samples and water suspensions of sediments (fresh water, brackish, and seawater sediments);
b) effluents (especially turbid and coloured);
c) aqueous extracts (e.g. leachates, eluates, elutriates) of soil, solid waste, and other solid material
(especially turbid and coloured).
2 Normative references
The following referenced documents are indispensable for the application 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 5667-16:1998, Water quality — Sampling — Part 16: Guidance on biotesting of samples
ISO 5814, Water quality — Determination of dissolved oxygen — Electrochemical probe method
ISO 11348-1, Water quality — Determination of the inhibitory effect of water samples on the light emission of
Vibrio fischeri (Luminescent bacteria test) — Part 1: Method using freshly prepared bacteria
ISO 11348-2, Water quality — Determination of the inhibitory effect of water samples on the light emission of
Vibrio fischeri (Luminescent bacteria test) — Part 2: Method using liquid-dried bacteria
ISO 11348-3:2007, Water quality — Determination of the inhibitory effect of water samples on the light
emission of Vibrio fischeri (Luminescent bacteria test) — Part 3: Method using freeze-dried bacteria
3 Terms and definitions
For the purpose of this document, the following terms and definitions apply.
3.1
contact time
duration of contact between one object or substance and another
NOTE In the test, the contact time is the time available to control or sample for contact with the test bacteria.
3.2
control sample
sample used in a laboratory in order to check or monitor the instrument or measurement performance or to
monitor changes in a sample under investigation
3.3
correction factor
dimensionless multiplier to correct data for known influences affecting their values as measured
NOTE In the test, the correction factor, f , serves to correct the initial luminescence intensity of the sample.
kt
3.4
peak value
maximum signal recorded in response to a stimulus
NOTE In the test, the peak value is the maximum signal which is recorded immediately after all the bacteria are in
contact with the sample.
3.5
reference sample
when the effect or behaviour of a substance is known from previous tests (reference substance) and when this
substance is examined within the framework of a test series as test sample, this is called the reference
sample
NOTE Adapted from ISO 5667-16:1998.
3.6
test sample
test sample is made from the sample by means of various preparatory steps specific to the sample and the
test, e.g. by dissolving, homogenizing, sedimenting, filtering, neutralizing or aeration
[ISO 5667-16:1998]
4 Principle
The inhibition of light emission by cultures of Vibrio fischeri is measured kinetically by following the light
emission of cultures from the very beginning of the assay. This is accomplished by dispensing the luminescent
bacteria suspension into the sample in a cuvette or other suitable vessel (e.g. microtiter plate) already in the
measuring position in the luminometer. The light emission is measured and recorded from the moment of
dispensing of the bacterial suspension to the sample until the maximum value has been reached and not only
at the maximum value of intensity (peak value) which usually occurs within 5 s of mixing, and after a contact
time of 15 min and 30 min or optionally 5 min (Figure 1).
2 © ISO 2010 – All rights reserved
Key
t time
y relative light units
1 start measurement
2 inject bacteria
3 record peak value from 0 s to 5 s
4 mix the sample before recording signal at 30 min
Figure 1 — Principal schematic protocol for the kinetic luminescent bacteria test
Vibrio fischeri suspension is dispensed and mixed into the sample in the measurement chamber of the
luminometer.
The test criterion is the decrease of the lumi
...
INTERNATIONAL ISO
STANDARD 21338
First edition
2010-07-15
Water quality — Kinetic determination of
the inhibitory effects of sediment, other
solids and coloured samples on the light
emission of Vibrio fischeri (kinetic
luminescent bacteria test)
Qualité de l'eau — Détermination cinétique des effets inhibiteurs des
échantillons de sédiment, autres solides et des échantillons colorés sur
la luminescence de Vibrio fischeri (essai cinétique de bactéries
luminescentes)
Reference number
©
ISO 2010
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
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
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.
© ISO 2010
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
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 copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2010 – All rights reserved
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Terms and definitions .2
4 Principle.2
5 Interferences .3
6 Reagents and materials .4
7 Apparatus.5
8 Sampling and sample pre-treatment .5
9 Procedure.6
10 Evaluation.7
11 Expression of results.9
12 Criteria of validity .11
13 Test report.11
Annex A (informative) Precision data .13
Annex B (informative) Typical kinetic curves from different samples .17
Annex C (informative) Dilution series .18
Bibliography.20
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.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
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.
ISO 21338 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
iv © ISO 2010 – All rights reserved
Introduction
The method specified in this International Standard is a kinetic modification of the luminescent bacteria test
specified in ISO 11348. The kinetic method overcomes the problems arising from intense colour and turbidity
in the samples. There is no need for sedimentation or centrifugation of turbid samples, or for the correction of
colour as described in ISO 11348.
This kinetic method uses luminometers capable of dispensing luminescent bacteria to the samples and
measuring the luminescent intensity over a period of time. In the method, the bacterial suspension is
dispensed and mixed with the sample in the measurement chamber of the luminometer. Several suitable
instruments are commercially available, but only a few of them are capable of cooling the measurement
chamber to (15 ± 1) °C as specified in ISO 11348. However, if the bacterial suspension and test samples are
kept at (15 ± 1) °C in a thermo-block before the measurement and during the whole incubation, the actual
temperature during the contact time is (15 ± 1) °C.
The measurements specified in this International Standard can be carried out using freshly prepared bacteria,
as well as freeze- or liquid-dried bacterial preparations. The various bacterial preparations can deliver different
results, especially in the presence of heavy metals (see ISO 11348). The laboratories responsible for the
results have the opportunity to select the most suitable bacterial preparation based on expert judgement and
information about the samples to be tested.
INTERNATIONAL STANDARD ISO 21338:2010(E)
Water quality — Kinetic determination of the inhibitory effects
of sediment, other solids and coloured samples on the light
emission of Vibrio fischeri (kinetic luminescent bacteria test)
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This International Standard does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this International
Standard be carried out by suitably trained staff.
1 Scope
This International Standard specifies the kinetic direct-contact method for determining the inhibitory effect of
suspensions of sediment and other solid samples, and also for problematic turbid or coloured aqueous
samples on the light emission of the marine bacterium Vibrio fischeri (NRRL B-11177).
This method is applicable to:
a) sediment samples and water suspensions of sediments (fresh water, brackish, and seawater sediments);
b) effluents (especially turbid and coloured);
c) aqueous extracts (e.g. leachates, eluates, elutriates) of soil, solid waste, and other solid material
(especially turbid and coloured).
2 Normative references
The following referenced documents are indispensable for the application 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 5667-16:1998, Water quality — Sampling — Part 16: Guidance on biotesting of samples
ISO 5814, Water quality — Determination of dissolved oxygen — Electrochemical probe method
ISO 11348-1, Water quality — Determination of the inhibitory effect of water samples on the light emission of
Vibrio fischeri (Luminescent bacteria test) — Part 1: Method using freshly prepared bacteria
ISO 11348-2, Water quality — Determination of the inhibitory effect of water samples on the light emission of
Vibrio fischeri (Luminescent bacteria test) — Part 2: Method using liquid-dried bacteria
ISO 11348-3:2007, Water quality — Determination of the inhibitory effect of water samples on the light
emission of Vibrio fischeri (Luminescent bacteria test) — Part 3: Method using freeze-dried bacteria
3 Terms and definitions
For the purpose of this document, the following terms and definitions apply.
3.1
contact time
duration of contact between one object or substance and another
NOTE In the test, the contact time is the time available to control or sample for contact with the test bacteria.
3.2
control sample
sample used in a laboratory in order to check or monitor the instrument or measurement performance or to
monitor changes in a sample under investigation
3.3
correction factor
dimensionless multiplier to correct data for known influences affecting their values as measured
NOTE In the test, the correction factor, f , serves to correct the initial luminescence intensity of the sample.
kt
3.4
peak value
maximum signal recorded in response to a stimulus
NOTE In the test, the peak value is the maximum signal which is recorded immediately after all the bacteria are in
contact with the sample.
3.5
reference sample
when the effect or behaviour of a substance is known from previous tests (reference substance) and when this
substance is examined within the framework of a test series as test sample, this is called the reference
sample
NOTE Adapted from ISO 5667-16:1998.
3.6
test sample
test sample is made from the sample by means of various preparatory steps specific to the sample and the
test, e.g. by dissolving, homogenizing, sedimenting, filtering, neutralizing or aeration
[ISO 5667-16:1998]
4 Principle
The inhibition of light emission by cultures of Vibrio fischeri is measured kinetically by following the light
emission of cultures from the very beginning of the assay. This is accomplished by dispensing the luminescent
bacteria suspension into the sample in a cuvette or other suitable vessel (e.g. microtiter plate) already in the
measuring position in the luminometer. The light emission is measured and recorded from the moment of
dispensing of the bacterial suspension to the sample until the maximum value has been reached and not only
at the maximum value of intensity (peak value) which usually occurs within 5 s of mixing, and after a contact
time of 15 min and 30 min or optionally 5 min (Figure 1).
2 © ISO 2010 – All rights reserved
Key
t time
y relative light units
1 start measurement
2 inject bacteria
3 record peak value from 0 s to 5 s
4 mix the sample before recording signal at 30 min
Figure 1 — Principal schematic protocol for the kinetic luminescent bacteria test
Vibrio fischeri suspension is dispensed and mixed into the sample in the measurement chamber of the
luminometer.
The test criterion is the decrease of the luminescence at each endpoint compared to the peak value, taking
into account a correction factor, f , which is measured from intensity changes of control samples during the
kt
exposure time. The inhibitory effect of the sample can be determined as the lowest ineffective dilution (LID)
value, or as effective concentration (EC or EC ) values by means of dilution series (e.g. as described in
20 50
Annex C). The LID value is the most concentrated test batch tested at which the inhibition of light emission is
<20 %. For higher levels of inhibition, the dilution effect relationship can be determined graphically or by
statistical analysis. The inhibition by a sample is expressed as the concentrations which results in 20 % and
50 % light reduction compared to the blank (EC and EC ). This value is interpolated within the dilution
20 50
series.
No extra correction procedures for colour and turbidity are needed because these factors remain the same
during the whole measurement. Inhibition at different contact times and different sample concentrations yields
complete kinetic toxicological data about the sample (inhibition, expressed as a percentage vs. concentration
vs. time) and enables assumptions to be made about the nature of the contaminants if compared with existing
data (see Annex B).
5 Inter
...
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