SIST EN 17359:2020
(Main)Stationary source emissions - Bioaerosols and biological agents - Sampling of bioaerosols and collection in liquids - Impingement method
Stationary source emissions - Bioaerosols and biological agents - Sampling of bioaerosols and collection in liquids - Impingement method
This Standard contains specifications for active sampling of bioaerosols from exhaust air flowing through a defined cross-section of a stack. It defines general principles that have to be taken into account during an isokinetic sampling campaign for bioaerosols by bubbling the exhaust air through a specific impinge designed for emission measurements.
In the Standard the application with culturable organisms is specified but the same principle might be applicable for other analysis methods (e.g. molecular and/or enzyme-based methods).
The impinger is designed to allow a sample volume flow of 1 m3/h to 1,8 m3/h, or 16 ℓ/min to 30 ℓ/min, respectively, and has been tested with regard to various microorganisms within broad concentration ranges
Emissionen aus stationären Quellen - Bioaerosole und biologische Agenzien - Probenahme von Bioaerosolen und Abscheidung in Flüssigkeiten - Impinger-Methode
Dieses Dokument enthält Festlegungen zur aktiven Probenahme von Bioaerosolen aus der Abluft, die durch einen definierten Querschnitt eines Abluftkanals strömt. Sie legt allgemeine Grundlagen fest, die bei einer isokinetischen Probenahmekampagne für Bioaerosole berücksichtigt werden müssen, wobei die Abluft durch einen spezifischen Impinger, der auf Emissionsmessungen ausgelegt ist, geleitet wird.
In diesem Dokument wird die Anwendung mit anzüchtbaren Organismen festgelegt, jedoch kann dasselbe Prinzip auch auf Verfahren angewendet werden, die nicht auf Kulturen basieren (z. B. molekulare und/oder enzymbasierte Verfahren).
Der Impinger ist auf einen Teilvolumenstrom von 1 m3/h bis 1,8 m3/h bzw. 16 l/min bis 30 l/min ausgelegt, und wurde für verschiedene Mikroorganismen in breiten Konzentrationsbereichen geprüft [1; 2; 3; 4].
Émissions de sources fixes - Bioaérosols et agents biologiques - Prélèvement des bioaérosols et collecte dans les liquides - Méthode d’impaction par bullage
Le présent document contient des spécifications relatives au prélèvement actif des bioaérosols présents dans l’air rejeté à travers une section définie d’une cheminée. Il définit des principes généraux qui doivent être pris en compte au cours d’une campagne de prélèvement isocinétique de bioaérosols, en faisant passer des bulles d’air rejeté à travers un bulleur spécifiquement conçu pour les mesurages d’émissions.
Bien que le présent document spécifie l’utilisation d’organismes cultivables, le même principe peut s’appliquer aux méthodes hors cultures (méthodes moléculaires et/ou de type enzymatique, par exemple).
Le bulleur est conçu pour obtenir un débit volumique d’échantillon de 1 m3/h à 1,8 m3/h, soit de 16 l/min à 30 l/min, et a été soumis à essai avec divers micro-organismes sur des gammes de concentrations étendues [1 ; 2 ; 3 ; 4].
Emisije nepremičnih virov - Bioaerosoli in biološki agensi - Vzorčenje bioaerosolov in zajem v tekočini - Metoda z izpiranjem
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 17359:2020
01-oktober-2020
Emisije nepremičnih virov - Bioaerosoli in biološki agensi - Vzorčenje
bioaerosolov in zajem v tekočini - Metoda z izpiranjem
Stationary source emissions - Bioaerosols and biological agents - Sampling of
bioaerosols and collection in liquids - Impingement method
Emissionen aus stationären Quellen - Bioaerosole und biologische Agenzien -
Probenahme von Bioaerosolen und Abscheidung in Flüssigkeiten - Impinger-Methode
Émissions de sources fixes - Bioaérosols et agents biologiques - Prélèvement des
bioaérosols et collecte dans les liquides - Méthode d’impaction par bullage
Ta slovenski standard je istoveten z: EN 17359:2020
ICS:
13.040.40 Emisije nepremičnih virov Stationary source emissions
SIST EN 17359:2020 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 17359:2020
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SIST EN 17359:2020
EN 17359
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2020
EUROPÄISCHE NORM
ICS 13.040.20
English Version
Stationary source emissions - Bioaerosols and biological
agents - Sampling of bioaerosols and collection in liquids -
Impingement method
Émissions de sources fixes - Bioaérosols et agents Emissionen aus stationären Quellen - Bioaerosole und
biologiques - Prélèvement des bioaérosols et collecte biologische Agenzien - Probenahme von Bioaerosolen
dans les liquides - Méthode d'impaction par bullage und Abscheidung in Flüssigkeiten - Impinger-Methode
This European Standard was approved by CEN on 5 July 2020.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17359:2020 E
worldwide for CEN national Members.
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SIST EN 17359:2020
EN 17359:2020 (E)
Contents Page
European foreword . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 7
4 Symbols and abbreviations . 11
5 Principle of method . 13
6 Theoretical fundamentals . 14
6.1 Isokinetic sampling . 14
6.2 Determination of concentration and load of the microorganisms . 14
7 Device and materials . 16
7.1 General . 16
7.2 Device and methods for measurement of the exhaust air parameters for the
calculation of the main volume flow . 16
7.2.1 General . 16
7.2.2 Device for determination of the exhaust air velocity . 16
7.2.3 Device for determination of pressure, temperature and humidity . 17
7.3 Device for the sampling of bioaerosols . 17
7.3.1 General . 17
7.3.2 Material properties . 18
7.3.3 Entry nozzle, bend and sampling probe . 18
7.3.4 Emission impinger . 18
7.3.5 Suction device and device for measurement of the gas volume or respectively the gas
volume flow . 20
8 Sampling . 20
8.1 General . 20
8.2 Preparation of the sampling equipment . 20
8.2.1 General . 20
8.2.2 Preparation of the emission impinger . 20
8.2.3 Preparation of the entry nozzle and the sampling probe . 21
8.2.4 Determination of appropriate sampling probe and sampling flow . 21
8.3 Performing bioaerosol sampling . 24
8.3.1 Leak test and sampling . 24
8.3.2 Recovery of deposits upstream of the emission impinger . 25
8.3.3 Determination of the mass of the sampling liquid . 26
8.3.4 Field blank value . 26
8.3.5 Analytical blank value . 27
8.4 Transport and storage . 27
9 Analysis . 27
10 Evaluation . 27
10.1 General . 27
10.2 Transfer of the results by the analytical laboratory . 28
10.3 Sample gas volume during sampling. 28
10.4 Microorganism number calculation . 30
10.5 Load calculation . 31
2
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11 Performance characteristerics . 31
11.1 Measurement uncertainty . 31
11.2 Parameters for the determination of measuring uncertainity in practice . 32
12 Maintenance and quality assurance . 37
13 Sampling efficiency and limits of the method . 37
14 Interferences . 38
Annex A (informative) Practical example for moulds and bacteria . 39
A.1 General . 39
A.2 Determination of the measurement points . 39
A.3 Devices and materials . 39
A.3.1 General . 39
A.3.2 Devices and methods for measurement of the exhaust air parameters for the
calculation of the main volume flow . 39
A.3.2.1 General . 39
A.3.2.2 Devices for determination of the exhaust air velocity . 39
A.3.2.3 Devices for determination of pressure, temperature and humidity. 40
A.3.3 Devices for sampling of bioaerosols . 40
A.3.3.1 General . 40
A.3.3.2 Material properties . 40
A.3.3.3 Entry nozzle, bend and sampling probe . 40
A.3.3.4 Emission impinger . 40
A.3.3.5 Measurement system for isokinetic sample volume flow abstraction . 40
A.4 Sampling process . 40
A.4.1 General . 40
A.4.2 Preparation of the sampling equipment . 40
A.4.2.1 General . 40
A.4.2.2 Preparation of the emission impinger . 41
A.4.2.3 Preparation of the entry nozzle, band and sampling probe . 41
A.4.3 Measurement of the exhaust air parameters for isokinetic sampling . 41
A.4.4 Sampling . 42
A.4.4.1 General . 42
A.4.4.2 Recovery of deposits upstream of the emission impinger . 42
A.4.4.3 Field blank value . 42
A.4.5 Transport and storage . 43
Annex B (informative) Measurement uncertainty . 49
B.1 General . 49
B.2 Determination of measurement uncertainty . 49
3
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EN 17359:2020 (E)
B.2.1 Moulds . 49
B.2.2 Mesophilic bacteria . 50
B.2.3 Total cell count . 50
B.2.4 Measurements in the bioaerosol test channel . 50
B.3 Field blank value . 51
Annex C (normative) Summary of the requirements to the emission measurement . 52
Annex D (informative) Sample protocol for sampling and analysis . 54
D.1 Sampling . 54
D.2 Analysis . 55
Bibliography . 56
4
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EN 17359:2020 (E)
European foreword
This document (EN 17359:2020) has been prepared by Technical Committee CEN/TC 264 “Air quality”,
the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by February 2021, and conflicting national standards shall
be withdrawn at the latest by February 2021.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia,
Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland,
Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of North
Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United
Kingdom.
5
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1 Scope
This document contains specifications for active sampling of bioaerosols from exhaust air flowing
through a defined cross-section of a stack. It defines general principles that have to be taken into account
during an isokinetic sampling campaign for bioaerosols by bubbling the exhaust air through a specific
impinger designed for emission measurements.
In this document the application with culturable organisms is specified but the same principle might be
applicable for non-cultural based methods (e.g. molecular and/or enzyme-based methods).
3 3
The impinger is designed to allow a sample volume flow of 1 m /h to 1,8 m /h, or 16 l/min to 30 l/min,
respectively, and has been tested with regard to various microorganisms within broad concentration
1
ranges [1; 2; 3; 4].
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN 1040:2005, Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of
basic bactericidal activity of chemical disinfectants and antiseptics - Test method and requirements
(phase 1)
EN 13284-1:2017, Stationary source emissions - Determination of low range mass concentration of dust -
Part 1: Manual gravimetric method
EN 15259:2007, Air quality - Measurement of stationary source emissions - Requirements for measurement
sections and sites and for the measurement objective, plan and report
CEN/TS 16115-1, Ambient air quality - Measurement of bioaerosols - Part 1: Determination of moulds using
filter sampling systems and culture-based analyses
EN ISO 16911-1, Stationary source emissions - Manual and automatic determination of velocity and volume
flow rate in ducts - Part 1: Manual reference method (ISO 16911-1)
EN ISO 20988:2007, Air quality - Guidelines for estimating measurement uncertainty (ISO 20988:2007)
1
This method is accepted by convention as reference method for determination of total emissions under application
of an out stack configuration according to EN 13284-1.
6
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EN 17359:2020 (E)
3 Terms and definitions
For the purposes of this document, the following terms and definitions.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https://www.iso.org/obp
— IEC Electropedia: available at http://www.electropedia.org/
3.1
waste gas
exhaust air
carrier gases with solid, liquid or gaseous constituents (emissions)
Note 1 to entry: The carrier gases can be natural air (e.g. from stable ventilation) or process gases.
3.2
analytical blank value
value determined by a blank sample covering the analytical procedure to ensure that no significant
contamination occurs during the complete analytical procedure
Note 1 to entry: according to CEN/TS 16115-1
3.3
bacteria
large group of prokaryotic microorganisms with one chromosome in a nuclear region and which replicate
only asexually by cell division
[SOURCE: EN 13098:2019, 3.3]
Note 1 to entry: The current classification of bacteria, i.e. the grouping to genera and species, is done according
to molecular sequence-based, chemotaxonomic and physiological properties.
3.4
standard conditions
reference values for a dry gas at a pressure of 101,325 kPa rounded to 101,3 kPa and a temperature of
273,15 K rounded to 273 K
Note 1 to entry: Where a regulatory authority stipulates reference conditions, for example through a site Licence
or Permit, this shall override the conditions specified above.
3.5
bioaerosol-
airborne particles of biological origin
[SOURCE: EN 13098:2019, 3.1]
Note 1 to entry: Bioaerosols in the sense of this European Standard are all aggregations of particles in the
atmosphere to which fungi (spores, conidia, fragments of hyphae), bacteria, viruses and/or pollen as well as their
cell membrane components and metabolites (e.g. endotoxins, mycotoxins) are attached or that consist of the above
mentioned components.
7
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3.6
biological sampling efficiency
capacity of the sampler to maintain the viability of the airborne microorganisms during collection and
also to keep the microbial products intact
[SOURCE: EN 13098:2019, 3.4]
Note 1 to entry: The biological sampling efficiency considers the sampling stress occurring during sampling and
analysis in addition to the physical sampling efficiency. It is strain- and species specific.
3.7
endotoxin
constituent of the external membrane of Gram-negative bacteria (lipopolysaccharide), consisting of a
complex lipid, lipid A, which is covalently bound to a polysaccharide
Note 1 to entry: "Free endotoxin" is liberated after cell death and by budding from living cells. Lipid A is the active
(toxic) part and is a potent pro-inflammatory substance and may induce febrile, bronchial and other symptoms in
exposed workers. The composition and the toxicity of endotoxin differs between species.
[SOURCE: EN 13098:2019, 3.9]
3.8
load
product of measured concentration of bioaerosols and the volumetric flow rate of the stack of the plant
3.9
overall blank sample
field blank sample
sample taken at the plant site in an identical manner to the normal samples in the series, except that the
sampling system is not inserted into the duct and no gas is sampled during test duration
Note 1 to entry: In this standard the term field blank sample is used.
3.10
total cell count
total number of viable and dead cells in a given volume
Note 1 to entry: The total cell count can be determined e.g. by DAPI staining.
Note 2 to entry: When for example stained with DNA fluorescent dyes like DAPI (4,6-Diamidino-2-phenylindole-
dihydrochloride), living and dead cells are jointly counted and cannot be distinguished.
3.11
main volume flow
volumetric flow of the flue gas in the sampling plane
3.12
impingement
separation of airborne particles in liquids by different mechanisms, i.e. impaction, diffusion, interception
and sedimentation
8
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3.13
Colony Forming Unit
CFU
unit by which the culturable number of microorganisms is expressed
[SOURCE: EN 13098:2019, 3.6 and 3.7]
Note 1 to entry: One Colony Forming Unit can originate from one single microorganism, an aggregate of many
microorganisms or from one or many microorganisms attached to one particle.
Note 2 to entry: The number of colonies which develop depends on cultivation conditions.
3.14
culture based analyses
cultivation
growing of microorganisms on culture media
[SOURCE: ISO 16000-16:2008, 3.6]
3.15
non-culture-based analyses
methods which do not rely on cultural methods including: microscopy, molecular methods, cytometry,
fluorescence etc.
3.16
indicator organisms
microorganisms which are characteristic for the emissions of a specific type of plant and can be detected
with currently available methods of sampling and analysis
Note 1 to entry: Indicator organisms which are characteristic of a specific source (process) can also occur in air
not influenced by the respective source, but mostly in minor concentrations. This is explained by the fact that many
microorganisms are found ubiquitously.
3.17
sampling line
measurement line
line in the sampling plane along which the sampling points are located, bounded by the inner duct wall
[SOURCE: EN 15259:2007, 3.15]
3.18
measurement site
place on the flue gas duct in the area of the sampling plane consisting of structures and technical
equipment, for example measurement platforms, measurement ports and energy supply
[SOURCE: EN 15259:2007, 3.11 – modified]
3.19
sampling point
measurement point
specific position on a sampling line at which a sample is extracted or the measurand directly determined
[SOURCE: EN 15259:2007, 3.16]
9
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3.20
sampling plane
measurement plane
plane normal to the centreline of the duct at the sampling position
[SOURCE: EN 15259:2007, 3.13]
Note 1 to entry: In the case of emission measurements of area sources, the total surface corresponds to the
sampling plane.
3.21
microbial air pollution
concentrations of airborne microorganisms that exceed natural concentrations or compositions which
differ in type from those occurring naturally
3.22
microorganism
microbial entity, either cellular or non cellular, that is capable of multiplication or transfer of genetic
material, or entities that have lost these properties
[SOURCE: EN 13098:2019, 3.18]
3.23
physical sampling efficiency
capacity of the sampling device to collect particles suspended in air
[SOURCE: EN 13098:2019, 3.20]
3.24
fungi
unicellular yeasts and filamentous fungi from several taxonomic groups namely zygomycetes
(Mucormycotina and Entomophthoromycotina), ascomycetes (Ascomycota) and mitosporic fungi
(deuteromycetes, fungi imperfecti) [7]
Note 1 to entry: Moulds form different types of spores depending on the taxonomic group they belong to, namely
conidiospores (conidia), sporangiospores or ascospores.
3.25
sample volumetric flow
volumetric flow taken off from the main stream for determination of the measured component
[SOURCE: EN 15259:2007, 3.29]
3.26
sampling train
fully assembled sampling system as per Figure 1
10
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4 Symbols and abbreviations
The symbols as outlined below are used throughout this standard:
Symbol Unit Meaning
A 2 sampling plane of the duct
m
A 2 area calculated on the basis of the diameter δ of the nozzle
S m S
c 3 3 number concentration of microorganisms in the sample flow
CFU/m ;TCC/m
C 3 3 number concentration of microorganisms in the exhaust air
CFU/m ;TCC/m
∑Cplate CFU sum of counted colonies on all evaluated plates
d dilution factor of the first evaluated dilution (for example d =
–
2
100 at a dilution degree 10 )
D additional dilution factor, which results from the fact that only
one aliquot of the dilutions has been outplated
3
f kg/m sample gas volume humidity at the volume meter (relative to
GM
the standard conditions)
3
f kg/m humidity in the exhaust air
n number of plates at the lowest evaluable dilution (higher
1
concentration)
n number of plates at the nearest higher evaluable dilution (lower
2
concentration)
N number of duplicate analyses
N CFU number of bacteria in cfu
CFU
p pipetting factor
p Pa dynamic pressure
d
p kPa absolute exhaust air pressure
G
p kPa absolute pressure at the gas volume meter
GM
p kPa standard pressure (101,3 kPa)
N
p kPa ambient air pressure
A
p kPa absolute pressure at the float-type flow meter inlet
R
p kPa static pressure in the duct
st
% volume fraction of water vapour in the humid exhaust air
r
H O, G
2
% volume fraction of water vapour in the humid exhaust air at the
r
H O, GM
2
gas volume meter
s 3 3 standard deviation under repeatability conditions
CFU/m ; TCC/m
T
...
SLOVENSKI STANDARD
oSIST prEN 17359:2019
01-marec-2019
[Not translated]
Stationary source emissions - Bioaerosols and biological agents - Sampling of
bioaerosols and collection in liquids - Impingement method
Emissionen aus stationären Quellen - Bioaerosole und biologische Agenzien -
Probenahme von Bioaerosolen und Abscheidung in Flüssigkeiten - Impinger-Methode
Ta slovenski standard je istoveten z: prEN 17359
ICS:
13.040.40 (PLVLMHQHSUHPLþQLKYLURY Stationary source emissions
oSIST prEN 17359:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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oSIST prEN 17359:2019
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oSIST prEN 17359:2019
DRAFT
EUROPEAN STANDARD
prEN 17359
NORME EUROPÉENNE
EUROPÄISCHE NORM
February 2019
ICS
English Version
Stationary source emissions - Bioaerosols and biological
agents - Sampling of bioaerosols and collection in liquids -
Impingement method
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 264.
If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations
which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.
This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other
language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC
Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17359:2019 E
worldwide for CEN national Members.
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oSIST prEN 17359:2019
prEN 17359:2019 (E)
Contents Page
European foreword . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Symbols and abbreviations . 10
5 Principle of method . 12
6 Theoretical fundamentals . 13
6.1 Isokinetic sampling . 13
6.2 Determination of concentration and load of the microorganisms . 14
7 Device and materials . 15
7.1 General . 15
7.2 Device and methods for measurement of the exhaust air parameters for the
calculation of the main volume flow . 15
7.2.1 General . 15
7.2.2 Device for determination of the exhaust air velocity . 15
7.2.3 Device for determination of pressure, temperature and humidity . 16
7.3 Device for the sampling of bioaerosols . 16
7.3.1 General . 16
7.3.2 Material properties . 17
7.3.3 Entry nozzle, bend and sampling probe . 17
7.3.4 Emission impinger . 17
7.3.5 Suction device and device for measurement of the gas volume or respectively the gas
volume flow . 19
8 Sampling . 19
8.1 General . 19
8.2 Preparation of the sampling equipment . 19
8.2.1 General . 19
8.2.2 Preparation of the emission impinger . 20
8.2.3 Preparation of the entry nozzle and the sampling probe . 20
8.2.4 Determination of appropriate sampling probe and sampling flow . 20
8.3 Performing bioaerosol sampling . 23
8.3.1 Leak test and sampling (E) . 23
8.3.2 Recovery of deposits upstream of the emission impinge . 24
8.3.3 Determination of the mass of the sampling liquid . 25
8.3.4 Field blank value . 25
8.3.5 Analytical blank value (taken at the analysis laboratory) . 25
8.4 Transport and storage . 26
9 Analysis . 26
10 Evaluation . 26
10.1 General . 26
10.2 Transfer of the results by the analytical laboratory . 26
10.3 Sample gas volume during sampling. 27
10.4 Microorganism number calculation . 29
2
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10.5 Load calculation . 29
11 Performance characteristerics . 30
11.1 Measurement uncertainty (see VDI 2100-6, chapter 5.1 and 5.2) . 30
11.2 Parameters for the determination of measuring uncertainity in practice (most text is
from VDI 2267-1, chapter 8.2.1 and chapter 8.2.2.1) . 30
12 Maintenance and quality assurance . 35
13 Sampling efficiency and limits of the method . 36
14 Interferences . 36
Annex A (informative) Practical example for moulds and bacteria . 37
A.1 General . 37
A.2 Determination of the measurement points . 37
A.3 Devices and materials . 37
A.3.1 General . 37
A.3.2 Devices and methods for measurement of the exhaust air parameters for the
calculation of the main volume flow . 37
A.3.2.1 General . 37
A.3.2.2 Devices for determination of the exhaust air velocity . 37
A.3.2.3 Devices for determination of pressure, temperature and humidity. 38
A.3.3 Devices for sampling of bioaerosols . 38
A.3.3.1 General . 38
A.3.3.2 Material properties . 38
A.3.3.3 Entry nozzle, bend and sampling probe . 38
A.3.3.4 Emission impinger . 38
A.3.3.5 Measurement system for isokinetic sample volume flow abstraction . 38
A.4 Sampling process . 38
A.4.1 General . 38
A.4.2 Preparation of the sampling equipment . 38
A.4.2.1 General . 38
A.4.2.2 Preparation of the emission impinger . 39
A.4.2.3 Preparation of the entry nozzle, band and sampling probe . 39
A.4.3 Measurement of the exhaust air parameters for isokinetic sampling . 39
A.4.4 Sampling . 39
A.4.4.1 General . 39
A.4.4.2 Recovery of deposits upstream of the emission impinger . 40
A.4.4.3 Field blank value . 40
A.4.5 Transport and storage . 40
Annex B (informative) . 44
B.1 Measurement uncertainty . 44
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B.2 Measurement uncertainty . 44
B.2.1 Moulds . 44
B.2.2 Mesophilic bacteria . 45
B.2.3 Total cell count . 45
B.2.4 Measurements in the bioaerosol test channel . 45
B.3 Field blank value . 45
Annex C (normative) Summary of the requirements to the emission measurement . 47
Annex D (informative) Sample protocol for sampling and analysis . 49
D.1 Sampling . 49
D.2 Analysis . 50
Bibliography . 51
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European foreword
This document (prEN 17359:2019) has been prepared by Technical Committee CEN/TC 264 “Air quality”,
the secretariat of which is held by DIN.
This document is currently submitted to the CEN Enquiry.
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1 Scope
This document contains specifications for active sampling of bioaerosols from exhaust air flowing
through a defined cross-section of a stack. It defines general principles that have to be taken into account
during an isokinetic sampling campaign for bioaerosols by bubbling the exhaust air through a specific
impinge designed for emission measurements.
In this document the application with culturable organisms is specified but the same principle might be
applicable for non-cultural based methods (e.g. molecular and/or enzyme-based methods).
3 3
The impinger is designed to allow a sample volume flow of 1 m /h to 1,8 m /h, or 16 l/min to 30 l/min,
respectively, and has been tested with regard to various microorganisms within broad concentration
1)
ranges [1; 2; 3; 4].
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN 1040:2005, Chemical disinfectants and antiseptics — Quantitative suspension test for the evaluation of
basic bactericidal activity of chemical disinfectants and antiseptics — Test method and requirements (phase
1)
EN 13284-1:2017, Stationary source emissions — Determination of low range mass concentration of dust
— Part 1: Manual gravimetric method
EN 15259:2007, Air quality — Measurement of stationary source emissions — Requirements for
measurement sections and sites and for the measurement objective, plan and report
CEN/TS 16115-1, Ambient air quality — Measurement of bioaerosols — Part 1: Determination of moulds
using filter sampling systems and culture-based analyses
EN ISO 16911-1, Stationary source emissions — Manual and automatic determination of velocity and
volume flow rate in ducts — Part 1: Manual reference method (ISO 16911-1)
EN ISO 20988:2007, Air quality — Guidelines for estimating measurement uncertainty (ISO 20988:2007)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at http://www.iso.org/obp
1)
This method is accepted by convention as reference method for determination of total emissions under application
of an out stack configuration according to EN 13284-1.
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3.1
waste gas
exhaust air
carrier gases with solid, liquid or gaseous constituents (emissions), the carrier gases can be natural air
(e.g., from stable ventilation) or process gases (e.g. from combustion processes)
3.2
aerodynamic diameter
3
diameter of a sphere of density 1 g/cm 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
[SOURCE: ISO 7708:1995, 2.2]
3.3
analytical blank value
value determined by a blank sample covering the analytical procedure to ensure that no significant
contamination occurs during the complete analytical procedure (according to CEN/TS 16115-1)
3.4
bacteria
large group of prokaryotic microorganisms with one chromosome in a nuclear region and which replicate
only asexually by cell division
[SOURCE: EN 13098:2000, 3.2]
Note 1 to entry: The current classification of bacteria, i.e. the grouping to genera and species, is done according
to molecular sequence-based, chemotaxonomic and physiological properties.
3.5
standard conditions
reference values for a dry gas at a pressure of 101,325 kPa rounded to 101,3 kPa and a temperature of
273,15 K rounded to 273 K. Where a regulatory authority stipulates reference conditions, for example
through a site Licence or Permit, this shall override the conditions specified above
3.6
bioaerosols
airborne particles of biological origin
[SOURCE: EN 13098:2000, 3.3]
Note 1 to entry: Bioaerosols in the sense of this European Standard are all aggregations of particles in the
atmosphere to which fungi (spores, conidia, fragments of hyphae), bacteria, viruses and/or pollen as well as their
cell membrane components and metabolites (e.g. endotoxins, mycotoxins) are attached or that consist of the above
mentioned components.
3.7
biological sampling efficiency
capacity of the sampler to maintain the viability of the airborne microorganisms during collection and
also to keep the microbial products intact
[SOURCE: EN 13098:2000, 3.4]
Note 1 to entry: The biological sampling efficiency considers the sampling stress occurring during sampling and
analysis in addition to the physical sampling efficiency. It is strain- and species specific.
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3.8
endotoxins
constituent of the external membrane of Gram-negative bacteria (lipopolysaccharide), consisting of a
complex lipid, lipid A, which is covalently bound to a polysaccharide
Note 1 to entry: "Free endotoxin" is liberated after cell death and by budding from living cells. Lipid A is the active
(toxic) part and is a potent pro-inflammatory substance and may induce febrile, bronchial and other symptoms in
exposed workers. The composition and the toxicity of endotoxin differs between species.
[SOURCE: EN 13098:2000, 3.8]
3.9
load
product of measured concentration of bioaerosols and the volumetric flow rate of the stack of the plant
3.10
overall blank sample
field blank sample
sample taken at the plant site in an identical manner to the normal samples in the series, except that the
sampling system is not inserted into the duct and no gas is sampled during test duration
Note 1 to entry: In this standard the term field blank sample is used.
3.11
total cell count
total number of viable and dead cells in a given volume; the total cell count can be determined e.g. by
DAPI staining
Note 1 to entry: When for example stained with DNA fluorescent dyes like DAPI (4,6-Diamidino-2-phenylindole-
dihydrochloride), living and dead cells are jointly counted and cannot be distinguished.
3.12
main volume flow
volumetric flow of the flue gas in the sampling plane
[SOURCE: EN ISO 16911-1]
3.13
impingement
separation of airborne particles in liquids by different mechanisms, i.e. impaction, diffusion, interception
and sedimentation
3.14
Colony Forming Unit
CFU
unit by which the culturable number of microorganisms is expressed
[SOURCE: EN 13098:2000, 3.5]
Note 1 to entry: One Colony Forming Unit can originate from one single microorganism, an aggregate of many
microorganisms or from one or many microorganisms attached to one particle.
Note 2 to entry: The number of colonies which develop depends on cultivation conditions.
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3.15
culture based analyses
growing of microorganisms on culture media
[SOURCE: ISO 16000-16:2008, 3,6]
3.16
non-culture based analyses
methods which do not rely on cultural methods including: microscopy, molecular methods, cytometry,
fluorescence etc.
3.17
indicator organism
microorganisms which are characteristic for the emissions of a specific type of plant and can be detected
with currently available methods of sampling and analysis; indicator organisms which are characteristic
of a specific source (process) can also occur in air not influenced by the respective source, but mostly in
minor concentrations; this is explained by the fact that many microorganisms are found ubiquitously
3.18
sampling line
measurement line
line in the sampling plane along which the sampling points are located, bounded by the inner duct wall
[SOURCE: EN 15259:2007, 3.15]
3.19
sampling site
measurement site
place on the flue gas duct in the area of the sampling plane consisting of structures and technical
equipment, for example measurement platforms, measurement ports and energy supply
[SOURCE: EN 15259:2007, 3.11]
3.20
sampling point
measurement point
specific position on a sampling line at which a sample is extracted or the measurand directly determined
[SOURCE: EN 15259:2007, 3.16]
3.21
sampling plane
measurement plane
plane normal to the centreline of the duct at the sampling position
[SOURCE: EN 15259:2007, 3.13]
Note 1 to entry: In the case of emission measurements of area sources, the total surface corresponds to the
sampling plane.
3.22
microbial air pollution
concentrations of airborne microorganisms that exceed natural concentrations or compositions which
differ in type from those occurring naturally
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3.23
microorganism
microbial entity, either cellular or non cellular, that is capable of multiplication or transfer of genetic
material, or entities that have lost these properties
[SOURCE: EN 13098:2000, 3.16]
3.24
physical sampling efficiency
capacity of the sampling device to collect particles suspended in air (adapted from EN 13098:2000, 3.17)
3.25
fungi
unicellular yeasts and filamentous fungi from several taxonomic groups namely zygomycetes
(Mucormycotina and Entomophthoromycotina), ascomycetes (Ascomycota) and mitosporic fungi
(deuteromycetes, fungi imperfecti) [7]
Note 1 to entry: Moulds form different types of spores depending on the taxonomic group they belong to, namely
conidiospores (conidia), sporangiospores or ascospores.
3.26
sample volumetric flow
volumetric flow taken off from the main stream for determination of the measured component
[SOURCE: EN 15259:2007, 3.29]
3.27
sampling train
fully assembled sampling system as per Figure 1
4 Symbols and abbreviations
The symbols as outlined below are used throughout this standard:
Symbol Unit Meaning
A 2 sampling plane of the duct
m
A 2 area calculated on the basis of the diameter δ of the nozzle
S m S
c 3 3 number concentration of microorganisms in the sample flow
CFU/m ;TCC/m
C 3 3 number concentration of microorganisms in the exhaust air
CFU/m ;TCC/m
∑Cplate CFU sum of counted colonies on all evaluated plates
d dilution factor of the first evaluated dilution (for example d = 100 at a
–
2
dilution degree 10 )
D additional dilution factor, which results from the fact that only one
aliquot of the dilutions has been outplated
3
f kg/m sample gas volume humidity at the volume meter (relative to the
GM
standard conditions)
3
f kg/m humidity in the exhaust air
n number of plates at the lowest evaluable dilution (higher
1
concentration)
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Symbol Unit Meaning
n number of plates at the nearest higher evaluable dilution (lower
2
concentration)
N number of duplicate analyses
N CFU number of bacteria in cfu
CFU
p pipetting factor
p Pa dynamic pressure
d
p kPa absolute exhaust air pressure
G
p kPa absolute pressure at the gas volume meter
GM
p kPa standard pressure (101,3 kPa)
N
p kPa ambient air pressure
A
p kPa absolute pressure at the float-type flow meter inlet
R
p kPa static pressure in the duct
st
% volume fraction of water vapour in the humid exhaust air
r
H O, G
2
% volume fraction of water vapour in the humid exhaust air at the gas
r
H O, GM
2
volume meter
s 3 3 standard deviation under repeatability conditions
CFU/m ; TCC/m
T K absolute exhaust air temperature
G
T K absolute temperature at the gas volume meter
GM
T K standard temperature (273 K)
N
T K temperature at the float-type flow meter inlet
R
u m/s flow velocity at the measurement point i
G,i
uaverage m/s average flow velocity within the measurement section
u m/s flow velocity in the entry nozzle
S
u uncertainty
v 3 sample gas volume
m
v ml volume of pitted aliquot
b
3
v m sample gas volume relative to the condition at the gas volume meter
GM
(temperature, pressure, humidity)
vN,tr 3 sample gas volume (relative to the standard conditions 273 K, 101,3
m
kPa and dry gas volume)
v ml volume of pipetting (plates 1-4)
p
v ml filtered volume in the impinger sampling liquid
1
v ml total volume of the impinger sampling liqui
...
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