SIST ISO 16000-16:2013
Indoor air - Part 16: Detection and enumeration of moulds - Sampling by filtration
Indoor air - Part 16: Detection and enumeration of moulds - Sampling by filtration
This part of ISO 16000 specifies requirements for long-term (0,5 h to several hours) sampling of moulds in indoor air by filtration. Following the instructions given, a sample is obtained for subsequent detection of moulds by cultivation after suspension according to ISO 16000-17, which is part of the complete measurement procedure. This part of ISO 16000 is not suitable for personal sampling.
Air intérieur - Partie 16: Détection et dénombrement des moisissures - Échantillonnage par filtration
L'ISO 16000-16:2008 sp�cifie les exigences d'�chantillonnage de longue dur�e (de 0,5 h � plusieurs heures) des moisissures dans l'air int�rieur par filtration. En suivant les instructions donn�es, un �chantillon est pr�lev� pour d�tection ult�rieure des moisissures par culture apr�s suspension, conform�ment � l'ISO 16000-17 qui fait partie du mode op�ratoire de mesurage complet.
L'ISO 16000-16:2008 ne convient pas pour l'�chantillonnage individuel.
Notranji zrak - 16. del: Ugotavljanje prisotnosti in števila gliv - Vzorčenje s filtriranjem
Ta del standarda ISO 16000 določa zahteve za dolgoročno (od 0,5 do nekaj ur) vzorčenje gliv v notranjem zraku s filtriranjem. Z upoštevanjem danih navodil se vzorec glede na standard ISO 16000-17, ki je del celotnega meritvenega postopka, pridobi za nadaljnje ugotavljanje prisotnosti gliv s kultivacijo. Ta del standarda ISO 16000 ni primeren za osebno vzorčenje.
General Information
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Standards Content (Sample)
INTERNATIONAL ISO
STANDARD 16000-16
First edition
2008-12-15
Indoor air —
Part 16:
Detection and enumeration of moulds —
Sampling by filtration
Air intérieur —
Partie 16: Détection et dénombrement des moisissures —
Échantillonnage par filtration
Reference number
ISO 16000-16:2008(E)
©
ISO 2008
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ISO 16000-16:2008(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
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.
COPYRIGHT PROTECTED DOCUMENT
© ISO 2008
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 2008 – All rights reserved
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ISO 16000-16:2008(E)
Contents Page
Foreword. iv
Introduction . vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions. 1
4 Principle. 3
5 Apparatus and materials. 3
5.1 Sampling device. 3
5.2 Materials . 4
6 Measurement procedure . 4
6.1 Preparation for sampling . 4
6.2 Sampling. 6
6.3 Sampling period. 6
6.4 Transport and storage. 6
7 Sampling efficiency and method limitations . 7
8 Calibration of flow rate, function control and maintenance of the sampling system . 7
8.1 Calibration of flow rate. 7
8.2 Function control and maintenance. 7
9 Quality assurance. 7
10 Sampling protocol . 8
11 Performance characteristics . 8
Annex A (informative) Recovery of spores on gelatine filters in combination
with polycarbonate filters . 9
Annex B (informative) Technical description of a suitable filtration device . 11
Annex C (informative) Sampling protocol. 14
Annex D (informative) Trials for method validation. 16
Bibliography . 20
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ISO 16000-16:2008(E)
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 16000-16 was prepared by Technical Committee ISO/TC 146, Air quality, Subcommittee SC 6, Indoor air.
ISO 16000 consists of the following parts, under the general title Indoor air:
⎯ Part 1: General aspects of sampling strategy
⎯ Part 2: Sampling strategy for formaldehyde
⎯ Part 3: Determination of formaldehyde and other carbonyl compounds — Active sampling method
⎯ Part 4: Determination of formaldehyde — Diffusive sampling method
⎯ Part 5: Sampling strategy for volatile organic compounds (VOCs)
⎯ Part 6: Determination of volatile organic compounds in indoor and test chamber air by active sampling on
®
Tenax TA sorbent, thermal desorption and gas chromatography using MS/FID
⎯ Part 7: Sampling strategy for determination of airborne asbestos fibre concentrations
⎯ Part 8: Determination of local mean ages of air in buildings for characterizing ventilation conditions
⎯ Part 9: Determination of the emission of volatile organic compounds from building products and
furnishing — Emission test chamber method
⎯ Part 10: Determination of the emission of volatile organic compounds from building products and
furnishing — Emission test cell method
⎯ Part 11: Determination of the emission of volatile organic compounds from building products and
furnishing — Sampling, storage of samples and preparation of test specimens
⎯ Part 12: Sampling strategy for polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins
(PCDDs), polychlorinated dibenzofurans (PCDFs) and polycyclic aromatic hydrocarbons (PAHs)
⎯ Part 13: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and
polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) — Collection on sorbent-backed filters
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ISO 16000-16:2008(E)
⎯ Part 14: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and
polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) — Extraction, clean-up and analysis by
high-resolution gas chromatography and mass spectrometry
⎯ Part 15: Sampling strategy for nitrogen dioxide (NO )
2
⎯ Part 16: Detection and enumeration of moulds — Sampling by filtration
⎯ Part 17: Detection and enumeration of moulds — Culture-based method
⎯ Part 23: Performance test for evaluating the reduction of formaldehyde concentrations by sorptive
building materials
⎯ Part 24: Performance test for evaluating the reduction of volatile organic compounds and carbonyl
compounds without formaldehyde concentrations by sorptive building materials
The following parts are under preparation:
⎯ Part 18: Detection and enumeration of moulds — Sampling by impaction
⎯ Part 19: Sampling strategy for moulds
⎯ Part 25: Determination of the emission of semi-volatile organic compounds by building products — Micro-
chamber method
⎯ Part 28: Sensory evaluation of emissions from building materials and products
The following parts are planned:
⎯ Part 20: Detection and enumeration of moulds — Sampling from house dust
⎯ Part 21: Detection and enumeration of moulds — Sampling from materials
⎯ Part 22: Detection and enumeration of moulds — Molecular methods
⎯ Part 27: Standard method for the quantitative analysis of asbestos fibres in settled dust
Furthermore,
⎯ ISO 12219-1 (under preparation), Indoor air — Road vehicles — Part 1: Whole vehicle test chamber —
Specification and method for the determination of volatile organic compounds in car interiors,
⎯ ISO 16017-1, Indoor, ambient and workplace air — Sampling and analysis of volatile organic compounds
by sorbent tube/thermal desorption/capillary gas chromatography — Part 1: Pumped sampling, and
⎯ ISO 16017-2, Indoor, ambient and workplace air — Sampling and analysis of volatile organic compounds
by sorbent tube/thermal desorption/capillary gas chromatography — Part 2: Diffusive sampling
focus on volatile organic compound (VOC) measurements.
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ISO 16000-16:2008(E)
Introduction
Mould is a common name for filamentous fungi from different taxonomic groups (Zygomycetes, Ascomycetes
[Ascomycota], Deuteromycetes). They form a mycelium (hyphae) and spores — namely conidiospores
(conidia), sporangiospores or ascospores — by which they become visible macroscopically. Most spores are
in the size range 2 µm to10 µm, some up to 30 µm and a very few up to 100 µm. Spores of some mould
genera are small and become airborne very easily (e.g. Aspergillus, Penicillium) while others are bigger
and/or embedded in a slime matrix (Stachybotrys, Fusarium) and less mobile.
Mould spores are widely distributed in the outdoor environment and, therefore, also occur in varying
concentrations indoors. Growth of moulds in indoor environments, however, should be considered a public
health problem because epidemiological studies have revealed that dampness and/or mould growth in homes
and health impairment of occupants are closely related.
Standardised methods for sampling, detection and enumeration of moulds including standards for sampling
strategies are important for comparative assessment of mould problems indoors. Before taking any
measurements, a measurement strategy is required.
[7]
The procedure specified in this part of ISO 16000 is based on VDI 4252-2 , which is widely used for
detection and enumeration of fungi in ambient air and was adapted to be suitable also for indoor environments.
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INTERNATIONAL STANDARD ISO 16000-16:2008(E)
Indoor air —
Part 16:
Detection and enumeration of moulds — Sampling by filtration
WARNING — The use of this part of ISO 16000 may involve hazardous materials, operations and
equipment. This part of ISO 16000 does not purport to address any safety problems associated with
its use. It is the responsibility of the user of this part of ISO 16000 to establish appropriate safety and
health practices and determine the applicability of regulatory limitations prior to use.
1 Scope
This part of ISO 16000 specifies requirements for long-term (0,5 h to several hours) sampling of moulds in
indoor air by filtration. Following the instructions given, a sample is obtained for subsequent detection of
moulds by cultivation after suspension according to ISO 16000-17, which is part of the complete measurement
procedure.
This part of ISO 16000 is not suitable for personal sampling.
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 16000-17, Indoor air — Part 17: Detection and enumeration of moulds — Culture-based method
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
aerodynamic diameter
diameter of a sphere of relative density 1 with the same terminal velocity due to gravitational force in calm air
as the particle, under the prevailing conditions of temperature, pressure and relative humidity
NOTE Adapted from ISO 7708:1995, 2.2.
3.2
biological preservation efficiency
capacity of the sampler to maintain the viability of the airborne microorganisms during collection and also to
keep the microbial products intact
[6]
[EN 13098:2000 ]
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ISO 16000-16:2008(E)
3.3
filamentous fungus
fungus growing in the form of filaments of cells known as hyphae
NOTE 1 Hyphae aggregated in bundles are called mycelia.
NOTE 2 The term “filamentous fungi” differentiates fungi with hyphal growth from yeasts.
3.4
filtration
collection of particles suspended in gas or liquid by flow through a porous medium
[6]
[EN 13098:2000 ]
NOTE In this part of ISO 16000, filtration is understood as the separation of microorganisms or moulds from a
defined volume of air by means of filters.
3.5
colony forming unit
cfu
unit by which the culturable number of microorganisms is expressed
[6]
[EN 13098:2000 ]
NOTE 1 One colony forming unit can originate from one single microorganism, from aggregates of many
microorganisms as well as from one or many microorganisms attached to a particle.
NOTE 2 The number of colonies can depend on the cultivation conditions.
3.6
cultivation
〈air quality〉 growing of microorganisms on culture media
3.7
field blank
〈air quality〉 sample taken in an identical manner as the real sample, but without sucking air through the
sampling apparatus
NOTE The resulting blank represents the number of cfu entering the sample simply by handling the filter during
sampling. The results of the field blanks are not used for correction of measurement results but to detect sampling errors
(see ISO 16000-17).
3.8
microorganism
any microbiological entity, cellular or non cellular, capable of replication or of transferring genetic material, or
entities that have lost these properties
[6]
[EN 13098:2000 ]
3.9
mould
〈air quality〉 filamentous fungi from several taxonomic groups namely Zygomycetes, Ascomycetes
(Ascomycota) and Deuteromycetes (fungi imperfecti)
NOTE Moulds form different types of spores depending on the taxonomic group they belong to, namely
conidiospores (conidia), sporangiospores or ascospores.
3.10
physical sampling efficiency
capacity of the sampler to collect particles with specific sizes suspended in air
[6]
[EN 13098:2000 ]
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ISO 16000-16:2008(E)
3.11
total sampling efficiency
product of the physical sampling efficiency and the biological preservation efficiency
[6]
[EN 13098:2000 ]
4 Principle
During filtration, a defined air quantity is sucked through a filter — on or in which separation of the suspended
particles occurs.
Airborne moulds are collected on gelatine filters resulting in a high total sampling efficiency (see Annex A).
Polycarbonate filters are used below the gelatine filters to enhance stability (see Annex A). Filters other than
those of gelatine may be used provided they have been shown to have a relative recovery of at least 90 % of
the mass recovered on the gelatine type.
The sampling device is constructed for the detection of particles of the size of mould spores (> 1 µm to
~30 µm). To achieve this, the flow velocity of the filter shall be in the range 100 mm/s to 250 mm/s.
3
NOTE 1 If a filter with a diameter of 80 mm is used, this flow velocity is achieved by a flow rate of about 1,5 m /h to
3
3,3 m /h (25 l/min to 55 l/min).
NOTE 2 This method has been validated for a flow velocity of 217 mm/s. The physical sampling efficiency for other
velocities may be lower.
NOTE 3 Particles > 30 µm are also retained by the filters. If the filter holder is operated in a hanging position (e.g.
outdoor measurements with strong winds or rain) it is nonetheless possible that bigger particles may not reach the filter
holder due to their inertia.
After sampling, the mould spores are cultivated and counted. This procedure is specified in ISO 16000-17.
5 Apparatus and materials
5.1 Sampling device
The following components are needed.
5.1.1 Stand, to position the sampling head at the sampling height needed.
5.1.2 Sampling head, to position the filter holder with the inserted filters in a hanging position, if necessary.
A bent pipe or hose connection can be used to connect the sampling head to the sampling apparatus. The
inner diameter of the pipe or hose shall be 8 mm to 10 mm.
5.1.3 Filter holder, sterile (disposable or sterilizable), to insert the filters.
1)
5.1.4 Filters, of gelatine , sterile, of pore size 3 µm, and of polycarbonate, sterile, of pore size 0,8 µm (see
Annex A).
5.1.5 Vacuum pump, ensuring a constant flow rate during continuous operation.
The flow rate has to be adjusted to produce a flow velocity at the filter in the range 100 mm/s to 250 mm/s
(see Clause 4).
1) Sartorius Stedim Biotech, Göttingen, is an example of a suitable commercial supplier. This information is given for the
convenience of users of this International Standard and does not constitute an endorsement by ISO of this supplier.
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ISO 16000-16:2008(E)
5.1.6 Gas volume meter, to determine the volume, in operating cubic metres, of gas sucked in at the
sampling head.
5.1.7 Timer, for presetting time and duration of sampling.
5.1.8 Protective housing, to protect the sampling device from harmful environmental conditions (optional,
mainly for outdoor use).
The distance between the upper edge of the protective housing and the lower edge of the sampling head
should be at least 400 mm.
5.2 Materials
5.2.1 Sterile container, for filter containment during transport; e.g. Petri dishes.
5.2.2 Insulated container, for sample transport.
5.2.3 Protective gloves, preferably sterile disposable gloves or disinfected.
5.2.4 Disinfectant, e.g. iso-propanol or ethanol (70 % volume fraction).
5.2.5 Sterile tweezers, to handle the filters.
6 Measurement procedure
6.1 Preparation for sampling
Assemble the sampling equipment according to Figure 1.
A detailed example of a suitable sampling device is given in Annex B.
Connect the gas volume meter between the pump and the sampling head in order to determine the sampling
air volume without any interference caused by the leakage flow rate of the pump. The volume of the sampled
3
air is displayed in operating cubic metres to an accuracy of 0,01 m . The temperature and pressure within the
gas volume meter and in the air are continuously monitored.
The sampling air volume in operating cubic metres referenced to air conditions, V , is calculated electronically
B
using Equation (1):
Tp
AG
VV= (1)
BG
Tp
GA
where
p is the air pressure;
A
p is the air pressure within the gas volume meter;
G
T is the air temperature;
A
T is the temperature within the gas volume meter;
G
V is the sampling air volume measured by the gas volume meter.
G
Prepare the required number of sampling devices in accordance with the measurement task and the
measurement strategy. The air flow rate of the sampling apparatus should not exceed 10 % of the air change
rate of the room in question. If the air change rate is unknown, the air volume sampled per hour shall not
exceed 10 % of the volume of the room.
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ISO 16000-16:2008(E)
Dimensions in metres
Key
1 filter holder with optional sampling head 5 vacuum pump p ambient air pressure
A
for suspension
2 gas volume meter (e.g. orifice plate, 6 timer p sample gas pressure
G
thermal mass flow rate meter)
a
3 electronic circuit for conversion into 7 filter for abraded material T ambient air
A
operating cubic metres temperature
4 display for sampling air volume in 8 optional protective housing against T sample gas
G
operating cubic metres adverse environmental conditions temperature
V sample gas volume
B
a
If a rotary vane vacuum pump is used; air flow shall be released horizontally.
Figure 1 — Schematic setup of the sampling device
NOTE 1 Low volume sampling can be used for small rooms, for example children’s rooms. High volume sampling may
be considered for very large indoor areas if the noise produced by the sampler is of no consequence, provided that the
constraints on sampling rate or air volume specified above are observed.
The fluctuation of the flow velocity during sampling should be not more than ± 2 %. During sampling, the flow
rate of the sampled air shall not be reduced by more than 10 % as a result of the increasing filter loading.
NOTE 2 There are self-regulating sampling systems to overcome these faults.
It is recommended to check the equipment for completeness and functionality with a check list.
Verify the calibration validity of the sampling device. Conduct new calibration, if necessary, prior to the
measurements (see Clause 8).
Use sterile filters and sterile filter holders for the measurements. Maintain filter sterility up to the moment of
sampling. Protect filters from dust, heat, and strong vibrations during transport.
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ISO 16000-16:2008(E)
6.2 Sampling
Sampling is usually conducted at a height of 0,75 m to 1,5 m above ground. For special purposes, other
heights might be applicable. Take care that no settled house dust is sucked into the sampling device when
sampling at low heights.
NOTE The orientation of the filter holder in indoor environments with only small air currents is of minor importance. A
hanging sampling head is only necessary in case of strong air movements (e.g. comparative measurements outdoors).
Mount the sterile filter holders containing the sterile filters on the sampling apparatus without any
contamination (preferably by using sterile disposable gloves). Inspect the filters in the filter holders visually for
integrity and exact, airtight fitting of the seat.
Start the sampling device in accordance with the manufacturer’s operating instructions.
Record the measurement task and measurement location in the sampling protocol (an example is given in
Annex C), as well as data concerning the exact positioning of the measurement apparatus, type of sampling
equipment, date and time of each individual sampling, sampling period, flow rate and, for ambient air
measurements, wind direction and wind velocity. Also record other parameters (e.g. temperature, relative
humidity) as well as specific circumstances, anomalies or interferences that may be relevant to the analyses
of the mould problem (e.g. perception of odour, type and location of possible additional emission sources).
After sampling, remove the filters and filter holders from the sampling apparatus using sterile or disinfected
gloves, and check the filters for integrity. Pack the filters in sealed containers (see 5.2) in order to avoid any
secondary contamination.
Multiple measurements using different sampling periods are recommended. This is especially important when
the level of the anticipated concentration of moulds is not known.
Take a minimum of one field blank for each measurement object preferably in the middle of the measurement
series. For this purpose, place a sterile filter holder with filter in the sampling head with the pump switched off,
then remove, pack and analytically process the filter in the same manner as the loaded filters. Avoid a
prolonged exposure of the filter to the surrounding air.
6.3 Sampling period
The sampling period is determined by the measurement task and the expected mould concentration. Usual
sampling times are 30 min to several hours.
6.4 Transport and storage
Protect loaded filters from disturbing influences (sunshine, humidity or desiccation, heat and dust, etc.), and
transport them to the laboratory immediately after sampling with the sampling side facing upwards in sealed
containers (see 5.2). The temperature during transport shall not exceed the incubation temperature,
(25 ± 3) °C. If necessary, cool samples during transport, but take care not to freeze them. Record the
conditions during transport (temperature, humidity, duration). Process samples, if possible, immediately, and
not later than 48 h after the end of the sampling period. Store samples in the laboratory at a temperature not
exceeding the incubation temperature.
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ISO 16000-16:2008(E)
7 Sampling efficiency and method limitations
The limitations of the method are determined by the physical and biological sampling efficiency.
Physical sampling efficiency of both gelatine and polycarbonate filters is in excess of 95 % for moulds with an
aerodynamic diameter range of > 1 µm using a flow velocity at the filter of v = 217 mm/s (Annex A).
The effect of desiccation — influencing the biological preservation efficiency — is not uniform, but depends on
temperature, relative humidity, sampling time at the moment of measuring, as well as the type of mould.
Fortunately, most mould spores are relatively insensitive towards desiccation, so that the filter method can
usually be successfully applied in this field. Validation trials have shown reduced recovery of Cladosporium
spp. during very hot and sunny weather (see Clause D.2).
8 Calibration of flow rate, function control and maintenance of the sampling system
8.1 Calibration of flow rate
Perform calibration of the sampling apparatus by means of a certified reference volume meter having a
measurement accuracy of not more than ± 2 % expressed in operational cubic metres, referenced to air
conditions. Connect the reference volume meter to the air inlet of the sampling apparatus. Make sure that the
air inlet orifice of the reference apparatus is unobstructed. After a successful adjustment of the flow rate,
check the display accuracy of the sampling apparatus against the reference volume meter. The air volume
sucked through the sampling apparatus for a duration of 60 min shall be indicated with an accuracy of ± 1 %
compared to the reference volume meter. The time after which a steady flow velocity is reached should not
exceed 5 s.
The usual verification of the flow rate (function control) depends on the stability of the apparatus. Carry out a
complete calibration prior to starting a new measurement programme or following significant changes, e.g.
when new or repaired equipment is used or after pump servicing. If the flow rate determined using the transfer
standard deviates more than 2 % from the value required for correct operation of the inlet, adjust the flow
controller according to the manufacturer's instructions.
8.2 Function control and maintenance
Carry out maintenance of mechanical parts of the sampling system (inlet and connecting pipe work) including
leak check according to the manufacturer’s instructions.
Check the overall method by determining field blanks (see also 6.2).
9 Quality assurance
It is important to ascertain the exact operating mode of the sampling device (lack of leaks/determination
...
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.ILOWULUDQMHPAir intérieur - Partie 16: Détection et dénombrement des moisissures - Échantillonnage par filtrationIndoor air - Part 16: Detection and enumeration of moulds - Sampling by filtration13.040.20Kakovost okoljskega zrakaAmbient atmospheresICS:Ta slovenski standard je istoveten z:ISO 16000-16:2008SIST ISO 16000-16:2013en,fr01-december-2013SIST ISO 16000-16:2013SLOVENSKI
STANDARD
SIST ISO 16000-16:2013
Reference numberISO 16000-16:2008(E)© ISO 2008
INTERNATIONAL STANDARD ISO16000-16First edition2008-12-15Indoor air — Part 16: Detection and enumeration of moulds — Sampling by filtration Air intérieur — Partie 16: Détection et dénombrement des moisissures — Échantillonnage par filtration
SIST ISO 16000-16:2013
ISO 16000-16:2008(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 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.
COPYRIGHT PROTECTED DOCUMENT
©
ISO 2008 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 2008 – All rights reserved
SIST ISO 16000-16:2013
ISO 16000-16:2008(E) © ISO 2008 – All rights reserved iiiContents Page Foreword.iv Introduction.vi 1 Scope.1 2 Normative references.1 3 Terms and definitions.1 4 Principle.3 5 Apparatus and materials.3 5.1 Sampling device.3 5.2 Materials.4 6 Measurement procedure.4 6.1 Preparation for sampling.4 6.2 Sampling.6 6.3 Sampling period.6 6.4 Transport and storage.6 7 Sampling efficiency and method limitations.7 8 Calibration of flow rate, function control and maintenance of the sampling system.7 8.1 Calibration of flow rate.7 8.2 Function control and maintenance.7 9 Quality assurance.7 10 Sampling protocol.8 11 Performance characteristics.8 Annex A (informative)
Recovery of spores on gelatine filters in combination with polycarbonate filters.9 Annex B (informative)
Technical description of a suitable filtration device.11 Annex C (informative)
Sampling protocol.14 Annex D (informative)
Trials for method validation.16 Bibliography.20
SIST ISO 16000-16:2013
ISO 16000-16:2008(E) iv © ISO 2008 – All rights reserved 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 16000-16 was prepared by Technical Committee ISO/TC 146, Air quality, Subcommittee SC 6, Indoor air. ISO 16000 consists of the following parts, under the general title Indoor air: ⎯ Part 1: General aspects of sampling strategy ⎯ Part 2: Sampling strategy for formaldehyde ⎯ Part 3: Determination of formaldehyde and other carbonyl compounds — Active sampling method ⎯ Part 4: Determination of formaldehyde — Diffusive sampling method ⎯ Part 5: Sampling strategy for volatile organic compounds (VOCs) ⎯ Part 6: Determination of volatile organic compounds in indoor and test chamber air by active sampling on Tenax TA® sorbent, thermal desorption and gas chromatography using MS/FID ⎯ Part 7: Sampling strategy for determination of airborne asbestos fibre concentrations ⎯ Part 8: Determination of local mean ages of air in buildings for characterizing ventilation conditions ⎯ Part 9: Determination of the emission of volatile organic compounds from building products and furnishing — Emission test chamber method ⎯ Part 10: Determination of the emission of volatile organic compounds from building products and furnishing — Emission test cell method ⎯ Part 11: Determination of the emission of volatile organic compounds from building products and furnishing — Sampling, storage of samples and preparation of test specimens ⎯ Part 12: Sampling strategy for polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polycyclic aromatic hydrocarbons (PAHs) ⎯ Part 13: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) — Collection on sorbent-backed filters SIST ISO 16000-16:2013
ISO 16000-16:2008(E) © ISO 2008 – All rights reserved v⎯ Part 14: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) — Extraction, clean-up and analysis by high-resolution gas chromatography and mass spectrometry ⎯ Part 15: Sampling strategy for nitrogen dioxide (NO2) ⎯ Part 16: Detection and enumeration of moulds — Sampling by filtration ⎯ Part 17: Detection and enumeration of moulds — Culture-based method ⎯ Part 23: Performance test for evaluating the reduction of formaldehyde concentrations by sorptive building materials ⎯ Part 24: Performance test for evaluating the reduction of volatile organic compounds and carbonyl compounds without formaldehyde concentrations by sorptive building materials The following parts are under preparation: ⎯ Part 18: Detection and enumeration of moulds — Sampling by impaction ⎯ Part 19: Sampling strategy for moulds ⎯ Part 25: Determination of the emission of semi-volatile organic compounds by building products — Micro-chamber method ⎯ Part 28: Sensory evaluation of emissions from building materials and products The following parts are planned: ⎯ Part 20: Detection and enumeration of moulds — Sampling from house dust ⎯ Part 21: Detection and enumeration of moulds — Sampling from materials ⎯ Part 22: Detection and enumeration of moulds — Molecular methods ⎯ Part 27: Standard method for the quantitative analysis of asbestos fibres in settled dust Furthermore, ⎯ ISO 12219-1 (under preparation), Indoor air — Road vehicles — Part 1: Whole vehicle test chamber — Specification and method for the determination of volatile organic compounds in car interiors, ⎯ ISO 16017-1, Indoor, ambient and workplace air — Sampling and analysis of volatile organic compounds by sorbent tube/thermal desorption/capillary gas chromatography — Part 1: Pumped sampling, and ⎯ ISO 16017-2, Indoor, ambient and workplace air — Sampling and analysis of volatile organic compounds by sorbent tube/thermal desorption/capillary gas chromatography — Part 2: Diffusive sampling focus on volatile organic compound (VOC) measurements.
SIST ISO 16000-16:2013
ISO 16000-16:2008(E) vi © ISO 2008 – All rights reserved Introduction Mould is a common name for filamentous fungi from different taxonomic groups (Zygomycetes, Ascomycetes [Ascomycota], Deuteromycetes). They form a mycelium (hyphae) and spores — namely conidiospores (conidia), sporangiospores or ascospores — by which they become visible macroscopically. Most spores are in the size range 2 µm to10 µm, some up to 30 µm and a very few up to 100 µm. Spores of some mould genera are small and become airborne very easily (e.g. Aspergillus, Penicillium) while others are bigger and/or embedded in a slime matrix (Stachybotrys, Fusarium) and less mobile. Mould spores are widely distributed in the outdoor environment and, therefore, also occur in varying concentrations indoors. Growth of moulds in indoor environments, however, should be considered a public health problem because epidemiological studies have revealed that dampness and/or mould growth in homes and health impairment of occupants are closely related. Standardised methods for sampling, detection and enumeration of moulds including standards for sampling strategies are important for comparative assessment of mould problems indoors. Before taking any measurements, a measurement strategy is required. The procedure specified in this part of ISO 16000 is based on VDI 4252-2 [7], which is widely used for detection and enumeration of fungi in ambient air and was adapted to be suitable also for indoor environments.
SIST ISO 16000-16:2013
INTERNATIONAL STANDARD ISO 16000-16:2008(E) © ISO 2008 – All rights reserved 1Indoor air — Part 16: Detection and enumeration of moulds — Sampling by filtration WARNING — The use of this part of ISO 16000 may involve hazardous materials, operations and equipment. This part of ISO 16000 does not purport to address any safety problems associated with its use. It is the responsibility of the user of this part of ISO 16000 to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 1 Scope This part of ISO 16000 specifies requirements for long-term (0,5 h to several hours) sampling of moulds in indoor air by filtration. Following the instructions given, a sample is obtained for subsequent detection of moulds by cultivation after suspension according to ISO 16000-17, which is part of the complete measurement procedure. This part of ISO 16000 is not suitable for personal sampling. 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 16000-17, Indoor air — Part 17: Detection and enumeration of moulds — Culture-based method 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 aerodynamic diameter diameter of a sphere of relative density 1 with the same terminal velocity due to gravitational force in calm air as the particle, under the prevailing conditions of temperature, pressure and relative humidity NOTE Adapted from ISO 7708:1995, 2.2. 3.2 biological preservation efficiency capacity of the sampler to maintain the viability of the airborne microorganisms during collection and also to keep the microbial products intact [EN 13098:2000 [6]] SIST ISO 16000-16:2013
ISO 16000-16:2008(E) 2 © ISO 2008 – All rights reserved 3.3 filamentous fungus fungus growing in the form of filaments of cells known as hyphae NOTE 1 Hyphae aggregated in bundles are called mycelia. NOTE 2 The term “filamentous fungi” differentiates fungi with hyphal growth from yeasts. 3.4 filtration collection of particles suspended in gas or liquid by flow through a porous medium [EN 13098:2000 [6]] NOTE In this part of ISO 16000, filtration is understood as the separation of microorganisms or moulds from a defined volume of air by means of filters. 3.5 colony forming unit cfu unit by which the culturable number of microorganisms is expressed [EN 13098:2000 [6]] NOTE 1 One colony forming unit can originate from one single microorganism, from aggregates of many microorganisms as well as from one or many microorganisms attached to a particle. NOTE 2 The number of colonies can depend on the cultivation conditions. 3.6 cultivation 〈air quality〉=growing of microorganisms on culture media 3.7 field blank 〈air quality〉 sample taken in an identical manner as the real sample, but without sucking air through the sampling apparatus NOTE The resulting blank represents the number of cfu entering the sample simply by handling the filter during sampling. The results of the field blanks are not used for correction of measurement results but to detect sampling errors (see ISO 16000-17). 3.8 microorganism any microbiological entity, cellular or non cellular, capable of replication or of transferring genetic material, or entities that have lost these properties [EN 13098:2000 [6]] 3.9 mould 〈air quality〉 filamentous fungi from several taxonomic groups namely Zygomycetes, Ascomycetes (Ascomycota) and Deuteromycetes (fungi imperfecti) NOTE Moulds form different types of spores depending on the taxonomic group they belong to, namely conidiospores (conidia), sporangiospores or ascospores. 3.10 physical sampling efficiency capacity of the sampler to collect particles with specific sizes suspended in air [EN 13098:2000 [6]] SIST ISO 16000-16:2013
ISO 16000-16:2008(E) © ISO 2008 – All rights reserved 33.11 total sampling efficiency product of the physical sampling efficiency and the biological preservation efficiency [EN 13098:2000 [6]] 4 Principle During filtration, a defined air quantity is sucked through a filter — on or in which separation of the suspended particles occurs. Airborne moulds are collected on gelatine filters resulting in a high total sampling efficiency (see Annex A). Polycarbonate filters are used below the gelatine filters to enhance stability (see Annex A). Filters other than those of gelatine may be used provided they have been shown to have a relative recovery of at least 90 % of the mass recovered on the gelatine type. The sampling device is constructed for the detection of particles of the size of mould spores (> 1 µm to ~30 µm). To achieve this, the flow velocity of the filter shall be in the range 100 mm/s to 250 mm/s. NOTE 1 If a filter with a diameter of 80 mm is used, this flow velocity is achieved by a flow rate of about 1,5 m3/h to 3,3 m3/h (25 l/min to 55 l/min). NOTE 2 This method has been validated for a flow velocity of 217 mm/s. The physical sampling efficiency for other velocities may be lower. NOTE 3 Particles > 30 µm are also retained by the filters. If the filter holder is operated in a hanging position (e.g. outdoor measurements with strong winds or rain) it is nonetheless possible that bigger particles may not reach the filter holder due to their inertia. After sampling, the mould spores are cultivated and counted. This procedure is specified in ISO 16000-17. 5 Apparatus and materials 5.1 Sampling device The following components are needed. 5.1.1 Stand, to position the sampling head at the sampling height needed. 5.1.2 Sampling head, to position the filter holder with the inserted filters in a hanging position, if necessary. A bent pipe or hose connection can be used to connect the sampling head to the sampling apparatus. The inner diameter of the pipe or hose shall be 8 mm to 10 mm. 5.1.3 Filter holder, sterile (disposable or sterilizable), to insert the filters. 5.1.4 Filters, of gelatine 1), sterile, of pore size 3 µm, and of polycarbonate, sterile, of pore size 0,8 µm (see Annex A). 5.1.5 Vacuum pump, ensuring a constant flow rate during continuous operation. The flow rate has to be adjusted to produce a flow velocity at the filter in the range 100 mm/s to 250 mm/s (see Clause 4).
1) Sartorius Stedim Biotech, Göttingen, is an example of a suitable commercial supplier. This information is given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of this supplier. SIST ISO 16000-16:2013
ISO 16000-16:2008(E) 4 © ISO 2008 – All rights reserved 5.1.6 Gas volume meter, to determine the volume, in operating cubic metres, of gas sucked in at the sampling head. 5.1.7 Timer, for presetting time and duration of sampling. 5.1.8 Protective housing, to protect the sampling device from harmful environmental conditions (optional, mainly for outdoor use). The distance between the upper edge of the protective housing and the lower edge of the sampling head should be at least 400 mm. 5.2 Materials 5.2.1 Sterile container, for filter containment during transport; e.g. Petri dishes. 5.2.2 Insulated container, for sample transport. 5.2.3 Protective gloves, preferably sterile disposable gloves or disinfected. 5.2.4 Disinfectant, e.g. iso-propanol or ethanol (70 % volume fraction). 5.2.5 Sterile tweezers, to handle the filters. 6 Measurement procedure 6.1 Preparation for sampling Assemble the sampling equipment according to Figure 1. A detailed example of a suitable sampling device is given in Annex B. Connect the gas volume meter between the pump and the sampling head in order to determine the sampling air volume without any interference caused by the leakage flow rate of the pump. The volume of the sampled air is displayed in operating cubic metres to an accuracy of 0,01 m3. The temperature and pressure within the gas volume meter and in the air are continuously monitored. The sampling air volume in operating cubic metres referenced to air conditions, VB, is calculated electronically using Equation (1): AGBGGATpVVTp= (1) where pA is the air pressure; pG is the air pressure within the gas volume meter; TA is the air temperature; TG is the temperature within the gas volume meter; VG is the sampling air volume measured by the gas volume meter. Prepare the required number of sampling devices in accordance with the measurement task and the measurement strategy. The air flow rate of the sampling apparatus should not exceed 10 % of the air change rate of the room in question. If the air change rate is unknown, the air volume sampled per hour shall not exceed 10 % of the volume of the room. SIST ISO 16000-16:2013
ISO 16000-16:2008(E) © ISO 2008 – All rights reserved 5Dimensions in metres
Key 1 filter holder with optional sampling head for suspension 5 vacuum pump pA ambient air pressure 2 gas volume meter (e.g. orifice plate, thermal mass flow rate meter) 6 timer pG sample gas pressure 3 electronic circuit for conversion into operating cubic metres 7 filter for abraded material a TA ambient air temperature 4 display for sampling air volume in operating cubic metres 8 optional protective housing against adverse environmental conditions TG sample gas temperature
VB sample gas volume a If a rotary vane vacuum pump is used; air flow shall be released horizontally. Figure 1 — Schematic setup of the sampling device
NOTE 1 Low volume sampling can be used for small rooms, for example children’s rooms. High volume sampling may be considered for very large indoor areas if the noise produced by the sampler is of no consequence, provided that the constraints on sampling rate or air volume specified above are observed. The fluctuation of the flow velocity during sampling should be not more than ± 2 %. During sampling, the flow rate of the sampled air shall not be reduced by more than 10 % as a result of the increasing filter loading. NOTE 2 There are self-regulating sampling systems to overcome these faults. It is recommended to check the equipment for completeness and functionality with a check list. Verify the calibration validity of the sampling device. Conduct new calibration, if necessary, prior to the measurements (see Clause 8). Use sterile filters and sterile filter holders for the measurements. Maintain filter sterility up to the moment of sampling. Protect filters from dust, heat, and strong vibrations during transport. SIST ISO 16000-16:2013
ISO 16000-16:2008(E) 6 © ISO 2008 – All rights reserved 6.2 Sampling Sampling is usually conducted at a height of 0,75 m to 1,5 m above ground. For special purposes, other heights might be applicable. Take care that no settled house dust is sucked into the sampling device when sampling at low heights. NOTE The orientation of the filter holder in indoor environments with only small air currents is of minor importance. A hanging sampling head is only necessary in case of strong air movements (e.g. comparative measurements outdoors). Mount the sterile filter holders containing the sterile filters on the sampling apparatus without any contamination (preferably by using sterile disposable gloves). Inspect the filters in the filter holders visually for integrity and exact, airtight fitting of the seat. Start the sampling device in accordance with the manufacturer’s operating instructions. Record the measurement task and measurement location in the sampling protocol (an example is given in Annex C), as well as data concerning the exact positioning of the measurement apparatus, type of sampling equipment, date and time of each individual sampling, sampling period, flow rate and, for ambient air measurements, wind direction and wind velocity. Also record other parameters (e.g. temperature, relative humidity) as well as specific circumstances, anomalies or interferences that may be relevant to the analyses of the mould problem (e.g. perception of odour, type and location of possible additional emission sources). After sampling, remove the filters and filter holders from the sampling apparatus using sterile or disinfected gloves, and check the filters for integrity. Pack the filters in sealed containers (see 5.2) in order to avoid any secondary contamination. Multiple measurements using different sampling periods are recommended. This is especially important when the level of the anticipated concentration of moulds is not known. Take a minimum of one field blank for each measurement object preferably in the middle of the measurement series. For this purpose, place a sterile filter holder with filter in the sampling head with the pump switched off, then remove, pack and analytically process the filter in the same manner as the loaded filters. Avoid a prolonged exposure of the filter to the surrounding air. 6.3 Sampling period The sampling period is determined by the measurement task and the expected mould concentration. Usual sampling times are 30 min to several hours. 6.4 Transport and storage Protect loaded filters from disturbing influences (sunshine, humidity or desiccation, heat and dust, etc.), and transport them to the laboratory immediately after sampling with the sampling side facing upwards in sealed containers (see 5.2). The temperature during transport shall not exceed the incubation temperature, (25 ± 3) °C. If necessary, cool samples during transport, but take care not to freeze them. Record the conditions during transport (temperature, humidity, duration). Process samples, if possible, immediately, and not later than 48 h after the end of the sampling period. Store samples in the laboratory at a temperature not exceeding the incubation temperature. SIST ISO 16000-16:2013
ISO 16000-16:2008(E) © ISO 2008 – All rights reserved 77 Sampling efficiency and method limitations The limitations of the method are determined by the physical and biological sampling efficiency. Physical sampling efficiency of both gelatine and polycarbonate filters is in excess of 95 % for moulds with an aerodynamic diameter range of > 1 µm using a flow velocity at the filter of v = 217 mm/s (Annex A). The effect of desiccation — influencing the biological preservation efficiency — is not uniform, but depends on temperature, relative humidity, sampling time at the moment of measuring, as well as the type of mould. Fortunately, most mould spores are relatively insensitive towards desiccation, so that the filter method can usually be successfully applied in this field. Validation trials have shown reduced recovery of Cladosporium spp. during very hot and sunny weather (see Clause D.2). 8 Calibration of flow rate, function control and maintenance of the sampling system 8.1 Calibration of flow rate Perform calibration of the sampling apparatus by means of a certified reference volume meter having a measurement accuracy of not more than ± 2 % expressed in operational cubic metres, referenced to air conditions. Connect the reference volume meter to the air inlet of the sampling apparatus. Make sure that the air inlet orifice of the reference apparatus is unobstructed. After a successful adjustment of the flow rate, check the display accuracy of the sampling apparatus against the reference volume meter. The air volume sucked through the sampling apparatus for a duration of 60 min shall be indicated with an accuracy of ± 1 % compared to the reference volume meter. The time after which a steady flow velocity is reached should not exceed 5 s. The usual verification of the flow rate (function control) depends on the stability of the apparatus. Carry out a complete calibration prior to starting a new measurement programme or following significant changes, e.g. when new or repaired equipment is used or after pump servicing. If the flow rate determined using the transfer standard deviates more than 2 % from the value required for correct operation of the inlet, adjust the flow controller according to the manufacturer's instructions. 8.2 Function control and maintenance Carry out maintenance of mechanical parts of the sampling system (inlet and connecting pipe work) including leak check according to the manufacturer’s instructions. Check the overall method by determining field blanks (see also 6.2). 9 Quality assurance It is important to ascertain the exact operating mode of the sampling device (lack of leaks/determination of the sampling volume). Additionally, give special attention to the pump and the sampling head equipped with filters. The laboratory shall implement quality assurance measures to be documented and made available any time (see ISO/IEC 17025 [5]). SIST ISO 16000-16:2013
ISO 16000-16:2008(E) 8 © ISO 2008 – All rights reserved 10 Sampling protocol Label the samples for unique identification.
...
NORME ISO
INTERNATIONALE 16000-16
Première édition
2008-12-15
Air intérieur —
Partie 16:
Détection et dénombrement des
moisissures — Échantillonnage par
filtration
Indoor air —
Part 16: Detection and enumeration of moulds — Sampling by filtration
Numéro de référence
ISO 16000-16:2008(F)
©
ISO 2008
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ISO 16000-16:2008(F)
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© ISO 2008
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quelque forme que ce soit et par aucun procédé, électronique ou mécanique, y compris la photocopie et les microfilms, sans l'accord écrit
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ii © ISO 2008 – Tous droits réservés
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ISO 16000-16:2008(F)
Sommaire Page
Avant-propos. iv
Introduction . vii
1 Domaine d'application. 1
2 Références normatives . 1
3 Termes et définitions. 1
4 Principe. 3
5 Appareillage et matériels . 3
5.1 Dispositif d'échantillonnage. 3
5.2 Matériels . 4
6 Mode opératoire de mesurage. 4
6.1 Préparation pour l'échantillonnage. 4
6.2 Échantillonnage . 6
6.3 Durée d'échantillonnage . 7
6.4 Transport et stockage . 7
7 Efficacité d'échantillonnage et limites de la méthode . 7
8 Étalonnage du débit, vérification du fonctionnement et entretien du système
d'échantillonnage. 7
8.1 Étalonnage du débit. 7
8.2 Vérification du fonctionnement et entretien . 8
9 Assurance qualité. 8
10 Protocole d'échantillonnage. 8
11 Caractéristiques de performance. 9
Annexe A (informative) Récupération des spores sur des filtres en gélatine associés à des filtres
en polycarbonate . 10
Annexe B (informative) Description technique d'un dispositif de filtration approprié . 12
Annexe C (informative) Protocole d'échantillonnage. 15
Annexe D (informative) Essais de validation de la méthode . 17
Bibliographie . 21
© ISO 2008 – Tous droits réservés iii
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ISO 16000-16:2008(F)
Avant-propos
L'ISO (Organisation internationale de normalisation) est une fédération mondiale d'organismes nationaux de
normalisation (comités membres de l'ISO). L'élaboration des Normes internationales est en général confiée
aux comités techniques de l'ISO. Chaque comité membre intéressé par une étude a le droit de faire partie du
comité technique créé à cet effet. Les organisations internationales, gouvernementales et non
gouvernementales, en liaison avec l'ISO participent également aux travaux. L'ISO collabore étroitement avec
la Commission électrotechnique internationale (CEI) en ce qui concerne la normalisation électrotechnique.
Les Normes internationales sont rédigées conformément aux règles données dans les Directives ISO/CEI,
Partie 2.
La tâche principale des comités techniques est d'élaborer les Normes internationales. Les projets de Normes
internationales adoptés par les comités techniques sont soumis aux comités membres pour vote. Leur
publication comme Normes internationales requiert l'approbation de 75 % au moins des comités membres
votants.
L'attention est appelée sur le fait que certains des éléments du présent document peuvent faire l'objet de
droits de propriété intellectuelle ou de droits analogues. L'ISO ne saurait être tenue pour responsable de ne
pas avoir identifié de tels droits de propriété et averti de leur existence.
L'ISO 16000-16 a été élaborée par le comité technique ISO/TC 146, Qualité de l'air, sous-comité SC 6, Air
intérieur.
L'ISO 16000 comprend les parties suivantes, présentées sous le titre général Air intérieur:
⎯ Partie 1: Aspects généraux de la stratégie d'échantillonnage
⎯ Partie 2: Stratégie d'échantillonnage du formaldéhyde
⎯ Partie 3: Dosage du formaldéhyde et d'autres composés carbonylés — Méthode par échantillonnage actif
⎯ Partie 4: Dosage du formaldéhyde — Méthode par échantillonnage diffusif
⎯ Partie 5: Stratégie d'échantillonnage pour les composés organiques volatils (COV)
⎯ Partie 6: Dosage des composés organiques volatils dans l'air intérieur des locaux et enceintes d'essai par
®
échantillonnage actif sur le sorbant Tenax TA , désorption thermique et chromatographie en phase
gazeuse utilisant MS/FID
⎯ Partie 7: Stratégie d'échantillonnage pour la détermination des concentrations en fibres d'amiante en
suspension dans l'air
⎯ Partie 8: Détermination des âges moyens locaux de l'air dans des bâtiments pour caractériser les
conditions de ventilation
⎯ Partie 9: Dosage de l'émission de composés organiques volatils de produits de construction et d'objets
d'équipement — Méthode de la chambre d'essai d'émission
⎯ Partie 10: Dosage de l'émission de composés organiques volatils de produits de construction et d'objets
d'équipement — Méthode de la cellule d'essai d'émission
iv © ISO 2008 – Tous droits réservés
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ISO 16000-16:2008(F)
⎯ Partie 11: Dosage de l'émission de composés organiques volatils de produits de construction et d'objets
d'équipement — Échantillonnage, conservation des échantillons et préparation d'échantillons pour essai
⎯ Partie 12: Stratégie d'échantillonnage des polychlorobiphényles (PCB), des polychlorodibenzo-p-dioxines
(PCDD), des polychlorodibenzofuranes (PCDF) et des hydrocarbures aromatiques polycycliques (HAP)
⎯ Partie 13: Dosage des polychlorobiphényles (PCB) de type dioxine et des polychlorodibenzo-p-dioxines
(PCDD)/polychlorodibenzofuranes (PCDF) totaux (en phase gazeuse et en phase particulaire) —
Collecte sur des filtres adsorbants
⎯ Partie 14: Dosage des polychlorobiphényles (PCB) de type dioxine et des polychlorodibenzo-p-dioxines
(PCDD)/polychlorodibenzofuranes (PCDF) totaux (en phase gazeuse et en phase particulaire) —
Extraction, purification et analyse par chromatographie en phase gazeuse haute résolution et
spectrométrie de masse
⎯ Partie 15: Stratégie d'échantillonnage du dioxyde d'azote (NO )
2
⎯ Partie 16: Détection et dénombrement des moisissures — Échantillonnage par filtration
⎯ Partie 17: Détection et dénombrement des moisissures — Méthode par culture
⎯ Partie 23: Essai de performance pour l'évaluation de la réduction des concentrations en formaldéhyde
par des matériaux de construction sorptifs
⎯ Partie 24: Essai de performance pour l'évaluation de la réduction des concentrations en composés
organiques volatils et en composés carbonyles sans formaldéhyde, par des matériaux de construction
sorptifs
Les parties suivantes sont en cours de préparation:
⎯ Partie 18: Détection et dénombrement des moisissures — Échantillonnage par impaction
⎯ Partie 19: Stratégie d'échantillonnage des moisissures
⎯ Partie 25: Détermination de l'émission de composés organiques semi-volatils des produits de
construction — Méthode de la micro-chambre
⎯ Partie 28: Évaluation sensorielle des émissions des matériaux et des produits de construction
Les parties suivantes sont prévues:
⎯ Partie 20: Détection et dénombrement des moisissures — Échantillonnage à partir de poussières
domestiques
⎯ Partie 21: Détection et dénombrement des moisissures — Échantillonnage à partir de matériaux
⎯ Partie 22: Détection et dénombrement des moisissures — Méthodes moléculaires
⎯ Partie 27: Méthode normalisée pour l'analyse quantitative des fibres d'amiante dans la poussière
En outre,
⎯ l'ISO 12219-1 (en préparation), Air intérieur — Véhicules routiers — Partie 1: Enceinte d'essai d'un
véhicule complet — Spécification et méthode de détermination des composés organiques volatils dans
les habitacles de voitures
© ISO 2008 – Tous droits réservés v
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ISO 16000-16:2008(F)
⎯ l'ISO 16017-1, Air intérieur, air ambiant et air des lieux de travail — Échantillonnage et analyse des
composés organiques volatils par tube à adsorption/désorption thermique/chromatographie en phase
gazeuse sur capillaire — Partie 1: Échantillonnage par pompage
⎯ l'ISO 16017-2, Air intérieur, air ambiant et air des lieux de travail — Échantillonnage et analyse des
composés organiques volatils par tube à adsorption/désorption thermique/chromatographie en phase
gazeuse sur capillaire — Partie 2: Échantillonnage par diffusion
traitent plus particulièrement des mesurages liés aux composés organiques volatils (COV).
vi © ISO 2008 – Tous droits réservés
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ISO 16000-16:2008(F)
Introduction
Le terme «moisissure» est le nom commun des champignons filamenteux appartenant à différents groupes
taxonomiques [Zygomycètes, Ascomycètes (Ascomycota) et Deutéromycètes]. Ils forment un mycélium
(hyphes) et des spores, à savoir des conidiospores (conidies), des sporangiospores ou des ascospores, qui
les rendent visibles à l'œil nu. La plupart des spores mesurent de 2 µm à 10 µm, certaines atteignent 30 µm et,
dans de rares cas, certaines peuvent mesurer jusqu'à 100 µm. Les spores de certains genres de moisissure
sont de petite taille et se mettent facilement en suspension dans l'air (par exemple, Aspergillus, Penicillium)
tandis que d'autres sont plus grandes et/ou intégrées à une matrice visqueuse (Stachybotrys, Fusarium), ce
qui les rend moins mobiles.
Les spores de moisissures sont disséminées un peu partout dans l'environnement extérieur et se retrouvent
ainsi en concentration variable à l'intérieur des bâtiments. Il convient toutefois de considérer la croissance des
moisissures dans les environnements intérieurs comme un problème d'hygiène. En effet, des études
épidémiologiques ont montré que l'humidité et/ou la croissance des moisissures dans les logements sont
étroitement liées aux problèmes de santé affectant les habitants.
L'existence de méthodes normalisées pour l'échantillonnage, la détection et le dénombrement des
moisissures, y compris des normes relatives à des stratégies d'échantillonnage, est importante pour
l'évaluation comparative des problèmes liés aux moisissures à l'intérieur des bâtiments. Avant de procéder à
tout mesurage, une stratégie d'échantillonnage est nécessaire.
[7]
Le mode opératoire spécifié dans la présente partie de l'ISO 16000 repose sur le guide VDI 4252-2 , qui est
couramment utilisé pour la détection et le dénombrement de champignons dans l'air ambiant et qui a été
adapté pour convenir également à des environnements intérieurs.
© ISO 2008 – Tous droits réservés vii
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NORME INTERNATIONALE ISO 16000-16:2008(F)
Air intérieur —
Partie 16:
Détection et dénombrement des moisissures —
Échantillonnage par filtration
AVERTISSEMENT — L'utilisation de la présente partie de l'ISO 16000 peut impliquer des matériaux,
des opérations et un équipement dangereux. La présente partie de l'ISO 16000 n'a pas pour but de
traiter tous les problèmes de sécurité qui sont liés à son utilisation. Il incombe à l'utilisateur d'établir
des pratiques appropriées en matière d'hygiène et de sécurité, et de déterminer l'applicabilité des
limites réglementaires avant son utilisation.
1 Domaine d'application
La présente partie de l'ISO 16000 spécifie les exigences d'échantillonnage de longue durée (de 0,5 h à
plusieurs heures) des moisissures dans l'air intérieur par filtration. En suivant les instructions données, un
échantillon est prélevé pour détection ultérieure des moisissures par culture après suspension, conformément
à l'ISO 16000-17 qui fait partie du mode opératoire de mesurage complet.
La présente partie de l'ISO 16000 ne convient pas pour l'échantillonnage individuel.
2 Références normatives
Les documents de référence suivants sont indispensables pour l'application du présent document. Pour les
références datées, seule l'édition citée s'applique. Pour les références non datées, la dernière édition du
document de référence s'applique (y compris les éventuels amendements).
ISO 16000-17, Air intérieur — Partie 17: Détection et dénombrement des moisissures — Méthode par culture
3 Termes et définitions
Pour les besoins du présent document, les termes et définitions suivants s'appliquent.
3.1
diamètre aérodynamique
diamètre d'une sphère de densité relative 1 possédant la même vitesse terminale de chute dans l'air calme
liée à la gravité que celle de la particule, dans les mêmes conditions de température, de pression et
d'humidité relative
NOTE Adapté de l'ISO 7708:1995, 2.2.
3.2
efficacité de conservation biologique
capacité de l'échantillonneur à conserver la viabilité des micro-organismes en suspension dans l'air pendant
la collecte et à conserver les produits microbiens intacts
[6]
[EN 13098:2000 ]
© ISO 2008 – Tous droits réservés 1
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ISO 16000-16:2008(F)
3.3
champignon filamenteux
champignon poussant sous la forme de filaments de cellules appelés hyphes
NOTE 1 Les hyphes agrégées en faisceaux sont appelées mycélia.
NOTE 2 Le terme «champignon filamenteux» distingue les champignons à hyphes des levures.
3.4
filtration
prélèvement de particules en suspension dans un gaz ou un liquide par passage à travers un milieu poreux
[6]
[EN 13098:2000 ]
NOTE Dans la présente partie de l'ISO 16000, la filtration désigne la séparation des micro-organismes ou des
moisissures d'un volume défini d'air au moyen de filtres.
3.5
unité formant colonie
ufc
unité dans laquelle est exprimé le nombre de micro-organismes cultivables
[6]
[EN 13098:2000 ]
NOTE 1 Une unité formant colonie peut provenir d'un seul micro-organisme, d'agrégats de nombreux micro-
organismes ou d'un ou plusieurs micro-organismes liés à une particule.
NOTE 2 Le nombre de colonies dépend des conditions de culture.
3.6
culture
〈qualité de l'air〉 croissance de micro-organismes sur des milieux de culture
3.7
blanc de site
〈qualité de l'air〉 échantillon prélevé de manière identique à un échantillon réel, mais sans aspirer d'air dans
l'appareillage d'échantillonnage
NOTE Le blanc obtenu représente le nombre d'ufc pénétrant dans l'échantillon simplement par manipulation du filtre
au cours de l'échantillonnage. Les résultats des blancs de site ne sont pas utilisés pour corriger les résultats de mesurage,
mais pour détecter des erreurs d'échantillonnage (voir l'ISO 16000-17).
3.8
micro-organisme
entité microbiologique, cellulaire ou non, capable de se reproduire ou de transférer du matériau génétique, ou
entité ayant perdu ces propriétés
[6]
[EN 13098:2000 ]
3.9
moisissure
〈qualité de l'air〉 champignon filamenteux appartenant à différents groupes taxonomiques, à savoir
Zygomycètes, Ascomycètes (Ascomycota) et Deutéromycètes (champignons imparfaits)
NOTE Les moisissures forment différents types de spores selon le groupe taxonomique auquel elles appartiennent, à
savoir des conidiospores (conidies), des sporangiospores ou des ascospores.
3.10
efficacité physique d'échantillonnage
capacité de l'échantillonneur à recueillir des particules de tailles spécifiques en suspension dans l'air
[6]
[EN 13098:2000 ]
2 © ISO 2008 – Tous droits réservés
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ISO 16000-16:2008(F)
3.11
efficacité totale d'échantillonnage
produit de l'efficacité physique d'échantillonnage par l'efficacité de conservation biologique
[6]
[EN 13098:2000 ]
4 Principe
Pendant la filtration, une quantité définie d'air est aspirée à travers un filtre, sur lequel ou dans lequel la
séparation des particules en suspension a lieu.
Les moisissures en suspension dans l'air sont prélevées sur des filtres en gélatine, ce qui permet d'atteindre
une efficacité totale d'échantillonnage élevée (voir Annexe A). Des filtres en polycarbonate sont utilisés sous
les filtres en gélatine afin d'accroître la stabilité (voir Annexe A). Il est possible d'utiliser des filtres autres que
ceux en gélatine, à condition d'avoir démontré qu'ils présentent un taux de récupération relative d'au moins
90 % par comparaison avec les filtres en gélatine.
Le dispositif d'échantillonnage est prévu pour détecter les particules de la taille des spores de moisissures
(> 1 µm et jusqu'à environ 30 µm). Pour ce faire, la vitesse d'écoulement sur le filtre doit être comprise entre
100 mm/s et 250 mm/s.
3
NOTE 1 Avec un filtre de 80 mm de diamètre, cette vitesse d'écoulement est atteinte avec un débit d'environ 1,5 m /h
3
à 3,3 m /h (25 l/min à 55 l/min).
NOTE 2 La présente méthode a été validée pour une vitesse d'écoulement de 217 mm/s. L'efficacité physique
d'échantillonnage peut être inférieure pour d'autres vitesses.
NOTE 3 Des particules > 30 µm sont également retenues par les filtres. Si le porte-filtre est utilisé dans une position
suspendue (par exemple, en cas de mesurages à l'extérieur en présence de vents violents ou de pluie), des particules
plus grosses peuvent toutefois ne pas atteindre le porte-filtre en raison de leur inertie.
Une fois prélevées, les spores de moisissures sont cultivées et dénombrées. Le mode opératoire est spécifié
dans l'ISO 16000-17.
5 Appareillage et matériels
5.1 Dispositif d'échantillonnage
Les éléments suivants sont nécessaires.
5.1.1 Support, pour placer la tête d'échantillonnage à la hauteur d'échantillonnage souhaitée.
5.1.2 Tête d'échantillonnage, pour suspendre, si nécessaire, le porte-filtre contenant les filtres.
Il est possible de relier la tête d'échantillonnage à l'appareillage d'échantillonnage à l'aide d'un tube coudé ou
d'un flexible. Le diamètre interne du tube ou du flexible doit être de 8 mm à 10 mm.
5.1.3 Porte-filtre, stérile (jetable ou stérilisable) pour introduire les filtres.
1)
5.1.4 Filtres, en gélatine , stériles, de 3 µm de dimension de pores et filtres en polycarbonate, stériles, de
0,8 µm de dimension de pores (voir Annexe A).
1) Sartorius Stedim Biotech, Göttingen, Allemagne, est un exemple de fournisseur approprié. Cette information est
donnée à l'intention des utilisateurs de la présente Norme internationale et ne signifie nullement que l'ISO approuve ou
recommande le fournisseur ainsi désigné.
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ISO 16000-16:2008(F)
5.1.5 Pompe à vide, pour assurer un débit constant tout au long de l'opération.
Le débit est à adapter afin d'atteindre une vitesse d'écoulement sur le filtre comprise entre 100 mm/s et
250 mm/s (voir Article 4).
5.1.6 Compteur à gaz, pour déterminer le volume de gaz aspiré au niveau de la tête d'échantillonnage, en
mètres cubes effectifs.
5.1.7 Minuterie, pour prérégler l'heure et la durée d'échantillonnage.
5.1.8 Boîtier de protection, pour protéger le dispositif d'échantillonnage des conditions environnementales
défavorables (facultatif, utile en particulier pour un usage en extérieur).
Il convient que la distance entre le bord supérieur du boîtier de protection et le bord inférieur de la tête
d'échantillonnage soit d'au moins 400 mm.
5.2 Matériels
5.2.1 Récipient stérile, pour le confinement pendant le transport. Par exemple, boîtes de Petri.
5.2.2 Récipient isolant, pour transporter l'échantillon.
5.2.3 Gants de protection, de préférence, gants stériles jetables ou désinfectés.
5.2.4 Désinfectant, par exemple, isopropanol ou éthanol (à 70 % en fraction volumique).
5.2.5 Pinces stériles, pour manipuler les filtres.
6 Mode opératoire de mesurage
6.1 Préparation pour l'échantillonnage
Assembler la ligne d'échantillonnage conformément à la Figure 1.
Un exemple détaillé de dispositif d'échantillonnage approprié est donné à l'Annexe B.
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ISO 16000-16:2008(F)
Dimensions en mètres
Légende
1 porte-filtre avec tête d'échantillonnage 5 pompe à vide p pressure barométrique
A
facultative pour suspension
2 compteur à gaz (par exemple, débitmètre 6 minuterie p pression du gaz
G
massique thermique à diaphragme) prélevé
a
3 circuit électronique pour la conversion en 7 filtre pour le produit d'abrasion T température de l'air
A
mètres cubes effectifs ambiant
4 écran d'affichage du volume d'air prélevé 8 boîtier de protection si nécessaire, en T température du gaz
G
en mètres cubes effectifs cas de conditions environnementales prélevé
défavorables
V volume de gaz prélevé
B
a
En cas d'utilisation d'une pompe à vide à palettes rotatives, l'air doit être émis horizontalement.
Figure 1 — Configuration schématique du dispositif d'échantillonnage
Raccorder le compteur de gaz entre la pompe et la tête d'échantillonnage afin de déterminer le volume d'air
prélevé sans interférence due au débit de fuite de la pompe. Le volume d'air prélevé s'affiche en mètres
3
cubes effectifs et avec une précision à 0,01 m près. La température et la pression à l'intérieur du compteur à
gaz et dans l'air sont enregistrées en continu.
Le volume d'air prélevé en mètres cubes effectifs par rapport aux conditions ambiantes, V , est calculé
B
électroniquement à l'aide de l'Équation (1):
Tp
AG
VV= (1)
BG
Tp
GA
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ISO 16000-16:2008(F)
où
p est la pression de l'air;
A
p est la pression de l'air à l'intérieur du compteur à gaz;
G
T est la température de l'air;
A
T est la température à l'intérieur du compteur à gaz;
G
V est le volume d'air prélevé mesuré par le compteur à gaz.
G
Préparer le nombre de dispositifs d'échantillonnage requis en fonction de la tâche de mesurage et de la
stratégie de mesurage. Il convient que le débit d'air du dispositif d'échantillonnage ne dépasse pas 10 % du
taux de renouvellement de l'air dans la pièce en question. Si le taux de renouvellement de l'air est inconnu, le
volume d'air prélevé par heure ne doit pas dépasser 10 % du volume de la pièce.
NOTE 1 Un prélèvement de faible volume peut être réalisé dans les petites pièces, telles que les chambres d'enfant.
Un prélèvement de grand volume peut être envisagé pour les très grands espaces intérieurs, à condition que le bruit émis
par l'échantillonneur soit sans conséquence et que les contraintes de débit d'échantillonnage et de volume d'air spécifiées
ci-dessus soient respectées.
Il convient que la fluctuation de la vitesse d'écoulement lors du prélèvement ne soit pas supérieure à ± 2 %.
Au cours de l'échantillonnage, la réduction du débit d'air prélevé en raison de l'encrassement du filtre ne doit
pas dépasser 10 %.
NOTE 2 Certains systèmes d'échantillonnage disposent d'une fonction de réglage automatique qui compense ce
problème.
Il est recommandé de vérifier que l'équipement est complet et fonctionnel au moyen d'une liste de contrôle.
Vérifier la validité de l'étalonnage du dispositif d'échantillonnage. Si nécessaire, procéder à un nouvel
étalonnage avant d'effectuer les mesurages (voir Article 8).
Réaliser les mesurages avec des filtres et des porte-filtres stériles. Garantir la stérilité des filtres jusqu'au
moment de l'échantillonnage. Protéger les filtres de la poussière, de la chaleur et des fortes vibrations au
cours du transport.
6.2 Échantillonnage
L'échantillonnage est généralement réalisé à une hauteur de 0,75 m à 1,5 m du sol. Dans certains cas,
d'autres hauteurs peuvent s'appliquer. Veiller à ne pas aspirer de poussière domestique sédimentée dans le
dispositif d'échantillonnage en cas d'échantillonnage à faible hauteur.
NOTE L'orientation du porte-filtre dans les environnements intérieurs avec peu de courants d'air est secondaire. La
suspension de la tête d'échantillonnage est nécessaire uniquement en cas de mouvements d'air importants (par exemple,
mesurages comparatifs en extérieur).
Monter les porte-filtres stériles contenant les filtres stériles sur l'appareillage d'échantillonnage sans
contamination (de préférence à l'aide de gants jetables stériles). Vérifier par inspection visuelle l'intégrité et
l'assise étanche à l'air des filtres placés dans les porte-filtres.
Mettre en marche le dispositif d'échantillonnage conformément au mode d'emploi du fabricant.
Noter la tâche et le lieu de mesurage dans le protocole d'échantillonnage (un exemple est donné dans
l'Annexe C), de même que les données relatives à la position exacte de l'appareillage de mesurage, le type
d'équipement d'échantillonnage, la date et l'heure de chaque échantillonnage individuel, la durée
d'échantillonnage, le débit et, pour les mesurages de l'air ambiant, la direction et la vitesse du vent. Consigner
également d'autres paramètres (tels que la température, l'humidité relative), ainsi que les circonstances
spécifiques, les anomalies et les interférences pouvant être pertinentes pour les analyses des moisissures
(par exemple, perception d'odeur, type et emplacement d'autres sources d'émissions possibles).
6 © ISO 2008 – Tous droits réservés
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ISO 16000-16:2008(F)
Après échantillonnage, retirer les filtres et les porte-filtres de l'appareillage d'échantillonnage à l'aide de gants
stériles ou désinfectés, et vérifier l'intégrité des filtres. Placer les filtres dans des récipients hermétiques
(voir 5.2) afin d'éviter toute contamination secondaire.
Il est recommandé de procéder à plusieurs mesurages avec des durées d'échantillonnage différentes. Ceci
est particulièrement important lorsque le niveau de concentration en moisissures attendu est inconnu.
Réaliser au moins un blanc de site pour chaque objet de mesurage, de préférence a
...
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