SIST EN 16868:2019
(Main)Ambient air - Sampling and analysis of airbone pollen grains and fungal spores for networks related to allergy - Volumetric Hirst method
Ambient air - Sampling and analysis of airbone pollen grains and fungal spores for networks related to allergy - Volumetric Hirst method
This document specifies the procedure to sample continuously and analyse the concentration of airborne pollen grains and fungal spores in ambient air using the volumetric Hirst type sampler. This document describes both the sampling and the analysis procedures for the purpose of allergy networks. Other specifications may be required. A synergy exists between pollen and fungal spores' exposition and air pollution which requests some specific and complementary information in order to decrease the risk of health impact.
Außenluft - Probenahme und Analyse luftgetragener Pollen und Pilzsporen für Allergienetzwerke - Volumetrische Hirst-Methode
Dieses Dokument legt das Verfahren fest zur kontinuierlichen Probenahme und zur Analyse der Konzentration luftgetragener Pollenkörner und Pilzsporen in der Umgebungsluft mittels eines volumetrischen Sammlers nach dem Hirst-Prinzip [1] [2] [3] (siehe Anhang A) oder einer gleichwertigen, vergleichbare Daten liefernden Methode.
Dieses Dokument beschreibt sowohl die Probenahme als auch die Analyseverfahren für die Zwecke von Allergienetzwerken. Für die anderen in der Einleitung erwähnten Aufgaben können andere Spezifikationen erforderlich sein.
Air ambiant - Échantillonnage et analyse des grains de pollen et des spores fongiques aériens pour les réseaux relatifs à l'allergie - Méthode volumétrique de Hirst
La présente Norme européenne spécifie le mode opératoire pour l’échantillonnage en continu et l’analyse de la concentration en grains de pollen et spores fongiques dans l’air ambiant en utilisant l’échantillonneur volumétrique de Hirst [1] [2] [3] (voir l’Annexe A) ou toute autre méthode équivalente garantissant des données comparables.
La présente Norme européenne décrit les modes opératoires d’échantillonnage et d’analyse aux fins des réseaux relatifs à l’allergie. Pour les autres objectifs mentionnés dans l’introduction, des spécifications différentes peuvent être requises.
Zunanji zrak - Vzorčenje in analiza cvetnega prahu in trosov gliv v zraku za alergijsko omrežje - Volumetrična Hirstova metoda
Ta dokument določa postopek za stalno vzorčenje in analiziranje koncentracije cvetnega prahu ter trosov gliv v zunanjem zraku z volumetričnim vzorčevalnikom tipa Hirst. Ta dokument opisuje postopke vzorčenja in tudi analize za namen alergijskih omrežij. Morda so potrebne druge specifikacije. Med izpostavljenostjo pelodnim zrnom in trosom gliv ter onesnaženostjo zraka obstaja sinergija, za katero so potrebne nekatere določene in dodatne informacije, da se lahko zmanjša tveganje za vpliv na zdravje.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 16868:2019
01-september-2019
Nadomešča:
SIST-TS CEN/TS 16868:2016
Zunanji zrak - Vzorčenje in analiza cvetnega prahu in trosov gliv v zraku za
alergijsko omrežje - Volumetrična Hirstova metoda
Ambient air - Sampling and analysis of airbone pollen grains and fungal spores for
networks related to allergy - Volumetric Hirst method
Außenluft - Probenahme und Analyse luftgetragener Pollen und Pilzsporen für
Allergienetzwerke - Volumetrische Hirst-Methode
Air ambiant - Échantillonnage et analyse des grains de pollen et des spores fongiques
aériens pour les réseaux relatifs à l'allergie - Méthode volumétrique de Hirst
Ta slovenski standard je istoveten z: EN 16868:2019
ICS:
13.040.20 Kakovost okoljskega zraka Ambient atmospheres
SIST EN 16868:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 16868:2019
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SIST EN 16868:2019
EN 16868
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2019
EUROPÄISCHE NORM
ICS 13.040.20 Supersedes CEN/TS 16868:2015
English Version
Ambient air - Sampling and analysis of airborne pollen
grains and fungal spores for networks related to allergy -
Volumetric Hirst method
Air ambiant - Échantillonnage et analyse des grains de Außenluft - Probenahme und Analyse luftgetragener
pollen en suspension dans l'air et des spores fongiques Pollen und Pilzsporen für Allergienetzwerke -
pour les réseaux relatifs à l'allergie - Méthode Volumetrische Hirst-Methode
volumétrique de Hirst
This European Standard was corrected and reissued by the CEN-CENELEC Management Centre on 12 June 2019.
This European Standard was approved by CEN on 8 March 2019.
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, North
Macedonia, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATIO N
E UR O P ÄISCHES KOMITEE FÜR NORMUN G
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. EN 16868:2019 E
worldwide for CEN national Members.
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EN 16868:2019 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 10
5 Sampling . 10
5.1 Equipment . 10
5.1.1 Apparatus . 10
5.1.2 Sampling support . 14
5.1.3 Installation conditions . 16
5.2 Operating procedure . 16
5.2.1 Preparation of the coating medium . 16
5.2.2 Support preparation . 17
5.2.3 Changing of the drum . 18
6 Analysis . 18
6.1 Equipment . 18
6.2 Operating procedure . 19
6.2.1 Support . 19
6.2.2 Mounting medium . 19
6.3 Methodology for counting . 19
6.3.1 Glass slide preparation for microscopy analysis for drum tape . 19
6.3.2 Optical microscopy . 21
6.3.3 Identification . 22
6.3.4 Counting method . 22
6.3.5 Data recording . 22
6.3.6 Conversion factor . 23
7 Performance characteristics for pollen and fungal spores counts . 24
7.1 General . 24
7.2 Integrated uncertainty assessment . 24
7.3 Uncertainty from counting error and counting routine. 24
7.4 Measurement uncertainty relating to sampling efficiency . 24
7.5 Measurement uncertainty relating to capture film, adhesive and specimen
preparation . 24
7.6 Measurement uncertainty relating to time discrimination . 25
7.7 Measurement uncertainty related to the detection limit . 25
7.8 Measurement uncertainty in relation to the calibration of the flow rate . 25
7.9 Measurement uncertainty relating to spatial representativity . 25
8 Quality assurance . 25
8.1 General . 25
8.2 Measurement site/trap . 25
8.2.1 Control . 25
8.2.2 Characterization of the site and its ambient conditions (passport of sampling site) . 25
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8.2.3 Spatial representativity . 26
8.3 Analyst . 26
8.4 Intra- and interlaboratory quality assessments . 26
8.4.1 General . 26
8.4.2 Repeatability . 26
8.4.3 Reproducibility and accuracy . 26
8.4.4 Sensitivity and specificity . 27
8.5 Network monitoring management . 27
Annex A (informative) Hirst type volumetric trap . 28
Annex B (informative) Pictures of impaction support . 29
Annex C (informative) Material Safety Data Sheets . 31
Annex D (informative) Identification key . 32
Bibliography . 37
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European foreword
This document (EN 16868:2019) 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 November 2019, and conflicting national standards shall
be withdrawn at the latest by November 2019.
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.
This document supersedes CEN/TS 16868:2015.
The main changes with respect to the previous edition are listed below:
a) the title has been changed;
b) modifications have been made to the Introduction, the Scope and Clauses 3, 4, 5 and 6;
c) new paragraphs have been added to Clauses 7 and 8;
d) modifications have been made to all Annexes;
e) Figures D.2 and D.3 have been modified;
f) the Bibliography has been readjusted;
g) editorial changes have been made.
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, 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
the United Kingdom.
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Introduction
Biological particles (pollen and fungal spores) are present in the air, causing health impacts at various
levels. In Europe, a lot of people suffer from pollinosis due to pollen and/or fungal spores (EFA, European
Federation of Allergy and Airways Diseases Patients Association, 2017). Pollen grains and fungal spores
are considered in some Member States as an air pollutant as well as particles suspended in the air
(PM ). In Europe, European Aerobiology Society (EAS) in coordination with International
10,2,5
Association for Aerobiology (IAA) manage the methodology of sampling, analysis, quality control,
development and information.
Persons and institutions involved in pollen forecasting have a scientific and public health responsibility.
A pollen forecast is a guideline for allergen avoidance with a direct influence on pollen allergy sufferers
and their behaviour. Pollen allergy sufferers are in need of such information since pollen allergy affects
their quality of life and pollen and spores are an abundant, environmental allergen. The health state of
pollen allergy sufferers should never be risked due to inadequate forecasts, financial interests or deficient
working routines applied in the fundamental work such as pollen data evaluation and all involved
processes (maintenance of the device, preparation, evaluation, handling and processing of data).
Further pollen data should be included in therapy (immunotherapy at least for one year) to objectify the
benefit of the personal therapy.
For the sampling and analysis of biological particles different methodology and operating procedures are
used.
Information on airborne pollen and spore concentration (counts and analyses) plays an important role
in aerobiology, as well as in other disciplines and fields of application, such as biodiversity, agriculture,
forestry, phytopathology, meteorology, climatology, paleo-ecology/-climatology, forensic science,
bioterrorism and health (sensitization and allergy). The method described in this European Standard is
aimed for the purposes of networks related to allergy. Besides, it may also be useful for other applications
mentioned above.
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1 Scope
This document specifies the procedure to sample continuously and to analyse the concentration of
airborne pollen grains and fungal spores in ambient air using the volumetric Hirst type sampler [1] [2]
[3] (see Annex A) or an even equivalent method assuring comparable data.
This document describes both the sampling and the analysis procedures for the purpose of networks
related to allergy. For the other tasks mentioned in the introduction, other specifications may be required.
2 Normative references
There are no normative references in this document.
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
NOTE For general terms, see [4] [5].
3.1
measurement accuracy
accuracy of measurement
accuracy
closeness of agreement between a measured quantity value and a true quantity value of a measurand
Note 1 to entry: The concept ‘measurement accuracy’ is not a quantity and is not given a numerical quantity
value. A measurement is said to be more accurate when it offers a smaller measurement error.
Note 2 to entry: The term “measurement accuracy” should not be used for measurement trueness and the term
“measurement precision” should not be used for ‘measurement accuracy’, which, however, is related to both these
concepts.
Note 3 to entry: Measurement accuracy is sometimes understood as closeness of agreement between measured
quantity values that are being attributed to the measurand.
[SOURCE: JCGM 200:2012]
3.2
clockwork
mechanism with a spring and toothed gearwheels, used to drive a mechanical clock, toy or other device
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3.3
combined standard measurement uncertainty
combined standard uncertainty
standard measurement uncertainty that is obtained using the individual standard measurement
uncertainties associated with the input quantities in a measurement model
Note 1 to entry: In case of correlations of input quantities in a measurement model, covariances must also be
taken into account when calculating the combined standard measurement uncertainty; see also ISO/IEC Guide
98-3:2014 [22].
3.4
defatted
surface conditions after clearing with a fat removing substance
3.5
drum
cylindrical device for the mounting of a sticky tape
3.6
exine
outer wall of pollen grain, also called an exosporium
3.7
eyepiece
lens or combination of lenses in an optical instrument through which the eye views the image formed by
the objective lens or lenses; ocular
3.8
flow meter
instrument for measuring the flow rate of a fluid in a pipe
3.9
flow rate
amount of fluid (air volume) that flows in a given time
3.10
fungal spore
sexual or asexual reproductive unit of fungi, capable of developing a new individual
3.11
hood
metal cover or canopy for a stove, ventilator, etc
3.12
impaction
sampling of airborne particles by inertial separation on any surface (e.g. of an adhesive)
3.13
magnetic stirrer
object or mechanical device used for stirring something
3.14
magnification
magnifying power of an instrument
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3.15
microscope
optical instrument having a magnifying lens or a combination of lenses for inspecting objects too small to
be seen or too small to be seen distinctly and in detail by the unaided eye
3.16
objective
optics (in a telescope, microscope, camera, or other optical system), the lens or combination of lenses,
that first receive the rays from the object and form the image in the focal plane of the eyepiece, as in a
microscope, or on a plate or screen as in a camera
Note 1 to entry: Also called object glass, object lens, objective lens.
3.17
orifice
opening or aperture, as of a tube or pipe; a mouthpiece with a slot-like opening on the side of the trap
3.18
particle
pollen and spores
3.19
pollen
male gametophyte of seed plants (either angiosperms or gymnosperms)
3.20
measurement precision
precision
closeness of agreement between indications or measured quantity values obtained by replicate
measurements on the same or similar objects under specified conditions
Note 1 to entry: Measurement precision is usually expressed numerically by measures of imprecision, such as
standard deviation, variance, or coefficient of variation under the specified conditions of measurement.
Note 2 to entry: The ‘specified conditions’ can be, for example, repeatability conditions of measurement,
intermediate precision conditions of measurement, or reproducibility conditions of measurement
(see ISO 5725-1:1994, [6]).
Note 3 to entry: Measurement precision is used to define measurement repeatability, intermediate measurement
precision, and measurement reproducibility.
Note 4 to entry: Sometimes “measurement precision” is erroneously used to mean measurement accuracy.
[SOURCE: JCGM 200:2012]
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3.21
repeatability condition of measurement
repeatability condition
condition of measurement, out of a set of conditions that includes the same measurement procedure,
same operators, same measuring system, same operating conditions and same location, and replicate
measurements on the same or similar objects over a short period of time
Note 1 to entry: A condition of measurement is a repeatability condition only with respect to a specified set of
repeatability conditions.
Note 2 to entry: In chemistry, the term “intra-serial precision condition of measurement” is sometimes used to
designate this concept.
[SOURCE: JCGM 200:2012]
3.22
reproducibility condition of measurement
reproducibility condition
condition of measurement, out of a set of conditions that includes different locations, operators,
measuring systems, and replicate measurements on the same or similar objects
Note 1 to entry: The different measuring systems may use different measurement procedures.
Note 2 to entry: A specification should give the conditions changed and unchanged, to the extent practical.
[SOURCE: JCGM 200:2012]
3.23
sensitivity
measurement of the proportion of search particle which is correctly identified
3.24
slide
rectangular piece of glass on which an object is mounted or placed for examination under a microscope
3.25
specificity
measurement of the proportion of non-searched particles which are correctly identified as different from
the searched particles
3.26
standard measurement uncertainty
standard uncertainty of measurement
standard uncertainty
measurement uncertainty expressed as a standard deviation
3.27
taxa
taxonomic groups of any rank, such as a species, genus, family or other rank
3.28
trap
sampling device
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3.29
vacuum pump
pump or device by which a partial vacuum can be produced
3.30
wind vane
mechanical device attached to an elevated structure; rotates freely depending on the direction of the wind
4 Principle
Ambient air is sampled by a volumetric suction system and directed towards a suitably coated sampling
surface through a specific orifice oriented towards the wind; the particles contained in the sampled air
are deposited by impaction on a continuously moving adhesive acceptor surface. The deposit on the
sampling surface is examined with an optical microscope in order to identify and count the pollen and
fungal spores per area (deposition rates). Using this method allows to calculate concentrations as a daily
mean or an hourly mean. The sampling is usually done at low-volume rate (10 l/min). It allows a
continuous sampling for up to seven days [7] [8] [9].
5 Sampling
5.1 Equipment
5.1.1 Apparatus
The sampling device and its functional principles are shown schematically in Figures 1, 2 and 3.
The complete sampling system (so called “trap”) containing the motor, the vacuum pump, the orifice, the
rotating drum, the wind vane, the clockwork system, the impaction support shall be:
— resistant to corrosion;
— well attached (i.e. resistant to wind-blow, etc.);
— always horizontal (at the head level).
The commercial devices that meet the requirements are presented in Annex A. For the different purposes,
refer to the specific publications.
The wind vane allows permanent rotation of the trap head so that the orifice faces the wind. The rain
shield ensures a weather protection for the orifice (i.e. rainfall).
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Key
1 wind vane
2 impact unit
3 rain shield
4 orifice (inlet)
5 screw for flow rate adjustment
6 vacuum pump
(Source: RNSA)
Figure 1 — Schematic figure and picture of a sampling device operating on the Hirst type
impactor – General view
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Key
1 orifice (inlet)
2 drum
3 clock
4 connection to vacuum pump
(Source: RNSA)
Figure 2 — Schematic figure of a sampling device operating on the Hirst type impactor –
Schematic view
5.1.1.1 Suction pump
The suction pump works 24 h a day and continuously throughout the year at the same flow rate. The
power supply may be either mains or battery driven (solar panels). The electric motor is capable of
continuous operation.
The suction system is, for instance, a vacuum pump. The flow rate of suction shall be regularly controlled
and adjusted accordingly.
The recommended flow rate is 10 l/min with a maximum permissible deviation of ± 10 % (±1 l/min).
The manufacturer shall ensure the flow specification and provide a flow verification and calibration
procedure that allows the user to ensure compliance with this specification throughout the life of the
trap. The validity of the calibration method recommended with an error of less than 10 % in the accuracy
of flow rate between calibration and operation shall be certified by the manufacturer.
As the accuracy of the calibration depends on the air resistance and characteristic curves of the individual
trap, vacuum pump and flow meter used [10], the validity has to be certified accordingly, i.e. specifically
for the trap/vacuum pump/flow meter combination delivered by the manufacturer.
The flow rate shall be checked at every change of the impaction support.
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Key
1 lid
2 start reference pointer
3 lock nut
4 orifice position
5 trapping surface
Figure 3 — The Hirst volumetric trap showing 7-day lid assembly with drum
5.1.1.2 Orifice (inlet) (Figure 2)
The orifice shall have the following dimensions (with associated tolerances):
— rectangular opening: 14 mm (±0,1 mm) × 2 mm (±0,1 mm);
— specific orifice length: from 19 mm to 25 mm;
— distance D from the inside orifice to the drum without the tape: 0,70 mm (±0,1 mm).
The depth allows the non-turbulence of laminar flow and directs the mixture of air and particles towards
the coated support. In consequence, an efficient particle impaction for pollen grains and fungal spores,
induced by the laminar flow, is ensured.
The distance D between orifice and drum shall be 0,70 mm (±0,1 mm) (see Figure 4 – distance D = A-B).
The distance D allows efficient particle impaction for pollen grains and fungal spores. It shall be controlled
[11] [12] [13].
The orifice should be directed into the air-stream using a wind vane.
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Key
1 drum
2 orifice
3 cover
A 20,5 mm or 22,5 mm, depending on supplier
B 19,8 mm or 21,8 mm, depending on supplier
C drum diameter 110 mm to 112 mm
D 0,7 mm (±0,1 mm)
Figure 4 — Pollen trap (Head of Hirst system)
5.1.2 Sampling support
5.1.2.1 General
Two possibilities are widely used depending on the requested sampling period as a sampling support:
— A glass slide for microscopy (76 mm × 26 mm) on which a transparent tape is fixed
(48 mm × 19 mm) reagents (see Figure B.1 in Annex B) for one
coated with specific day of sampling.
— A drum (110 mm to 112 mm in diameter) on which a transparent coated flexible tape is attached for
seven days of sampling (see Figure B.2 in Annex B). The length of this tape ranges from 345 mm to
350 mm (±0,5 mm) depending on the size of the drum.
The sampling support is driven by a clockwork with a scrolling speed ranging from 2 mm/h to 14 mm/h
(±0,01 mm/h) depending on the sampling period.
The sampling support shall scroll regularly in front of the back outlet of the orifice. Sampling shall always
be continuous and stable and not be stopped during the requested sampling period.
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5.1.2.2 Transparent tape
The transparent tape is coated with an adhesive in order to fix the particles.
The following requirements shall be fulfilled:
— The transparent tape shall be not hygroscopic.
— The thickness of the whole transparent tape shall not be changed over time, and should not be
affected by operational conditions (temperature between –20 °C to +60 °C or humidity between
20 % and 100 %).
— It shall be transparent to allow the passing of microscopic light.
— The length shall be adapted to the support used (see require
...
SLOVENSKI STANDARD
oSIST prEN 16868:2017
01-november-2017
=XQDQML]UDN9]RUþHQMHLQDQDOL]DFYHWQHJDSUDKXLQWURVRYJOLYY]UDNX]D
DOHUJLMVNRRPUHåMH9ROXPHWULþQD+LUVWRYDPHWRGD
Ambient air - Sampling and analysis of airborne pollen grains and fungal spores for
allergy networks - Volumetric Hirst method
Außenluft - Probenahme und Analyse luftgetragener Pollen und Pilzsporen für
Allergienetzwerke - Volumetrische Hirst-Methode
Air ambiant - Échantillonnage et analyse des grains de pollen et des spores fongiques
aériens pour les réseaux relatifs à l'allergie - Méthode volumétrique de Hirst
Ta slovenski standard je istoveten z: prEN 16868
ICS:
13.040.20 Kakovost okoljskega zraka Ambient atmospheres
oSIST prEN 16868:2017 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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oSIST prEN 16868:2017
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oSIST prEN 16868:2017
DRAFT
EUROPEAN STANDARD
prEN 16868
NORME EUROPÉENNE
EUROPÄISCHE NORM
September 2017
ICS 13.040.20 Will supersede CEN/TS 16868:2015
English Version
Ambient air - Sampling and analysis of airbone pollen
grains and fungal spores for networks related to allergy -
Volumetric Hirst method
Air ambiant - Échantillonnage et analyse des grains de Außenluft - Probenahme und Analyse luftgetragener
pollen et des spores fongiques aériens pour les réseaux Pollen und Pilzsporen für Allergienetzwerke -
relatifs à l'allergie - Méthode volumétrique de Hirst Volumetrische Hirst-Methode
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: Avenue Marnix 17, B-1000 Brussels
© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 16868:2017 E
worldwide for CEN national Members.
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oSIST prEN 16868:2017
prEN 16868:2017 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 9
5 Sampling . 9
5.1 Equipment . 9
5.1.1 Apparatus . 9
5.1.2 Sampling support . 12
5.1.3 Wind vane and rain shield . 14
5.1.4 Complete sampling system . 15
5.2 Operating procedure . 15
5.2.1 Preparation of the coating medium . 15
5.2.2 Support preparation . 16
5.2.3 Changing of the drum . 17
6 Analysis . 17
6.1 Equipment . 17
6.2 Operating procedure . 18
6.2.1 Support . 18
6.2.2 Mounting medium . 18
6.3 Methodology for counting . 19
6.3.1 Glass slide preparation for microscopy analysis for drum tape . 19
6.3.2 Optical microscopy [14] [16] . 21
6.3.3 Identification [15] [16] . 21
6.3.4 Counting method . 21
6.3.5 Data recording . 21
6.3.6 Conversion factor . 22
7 Performance characteristics for pollen and fungal spores counts [10] [11] [17] [18] . 23
7.1 General . 23
7.2 Integrated uncertainty assessment . 23
7.3 Uncertainty from counting error and counting routine. 23
7.4 Measurement uncertainty relating to sampling efficiency . 24
7.5 Measurement uncertainty relating to capture film, adhesive and specimen
preparation . 24
7.6 Measurement uncertainty relating to time discrimination . 24
7.7 Measurement relating to the detection limit and dealing with values below the
detection limit . 24
7.8 Measurement uncertainty in relation to the calibration of the flow rate . 24
7.9 Measurement uncertainty relating to spatial representativity . 24
8 Quality assurance . 24
8.1 General . 24
8.2 Measurement site/trap . 25
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8.2.1 Control . 25
8.2.2 Characterization of the site and its ambient conditions (passport of sampling site) . 25
8.2.3 Spatial representativity . 25
8.3 Analyst . 25
8.4 Intra- and interlaboratory quality assessments . 25
8.4.1 General . 25
8.4.2 Repeatability . 25
8.4.3 Reproducibility and accuracy . 26
8.4.4 Sensitivity and specificity . 26
8.5 Network monitoring management . 26
Annex A (informative) Hirst type volumetric trap . 28
Annex B (informative) . 29
Pictures of impaction support
Annex C (informative) Material Safety Data Sheets. 31
Annex D (informative) Key of determination . 32
Bibliography . 36
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European foreword
This document (prEN 16868:2017) 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.
This document will supersede CEN/TS 16868:2015.
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Introduction
Biological particles (pollen and fungal spores) are present in the air, causing health impacts at various
levels. In Europe, nearly 18 % to 20 % of people suffer from pollinosis due to pollen and/or fungal spores.
well as
Pollen grains and fungal spores are considered in some Member States as an air pollutant as
particles suspended in the air (PM , ). In Europe, European Aerobiology Society (EAS) in
10 2,5 coordination
with International Association for Aerobiology (IAA) manage problems of sampling, analysis, quality
control, development and information.
Persons and institutions involved in pollen forecasting have a scientific and public health responsibility.
A pollen forecast is a guideline for allergen avoidance with a direct influence on pollen allergy sufferers
and their behavior. Pollen allergy sufferers are in need of such information since pollen allergy affects
their quality of life and pollen and spores are an abundant, environmental allergen. The health state of
pollen allergy sufferers should never be risked due to inadequate forecasts, financial interests or deficient
working routines applied in the fundamental work such as pollen data evaluation and all involved
processes (maintenance of the device, preparation, evaluation, handling and processing of data).
For the sampling and analysis of biological particles different methodology and operating procedures are
used.
Sampling equipment is diversified (see Annex A). Analysis is based on optical light microscopy for
identification and counting pollen grains and fungal spores.
Elements and reagents used during sampling and analysis have very specific properties and require to be
handled carefully.
Information on airborne pollen and spore concentration (counts and analyses) plays an important role
in aerobiology, as well as in other disciplines and fields of application, such as biodiversity, agriculture,
forestry, phytopathology, meteorology, climatology, paleo-ecology/-climatology, forensic science,
bioterrorism and health (sensitization and allergy). The method described in this European Standard is
aimed for the purposes of networks related to allergy. Besides, it may also be useful for other applications
mentioned above.
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1 Scope
This European Standard specifies the procedure to sample continuously and analyses the concentration
of airborne pollen grains and fungal spores in ambient air using the volumetric Hirst type sampler [1] [2]
[3] (see Annex A) or even equivalent method assuring comparable data.
This European Standard describes both the sampling and the analysis procedures for the purpose of
networks related to allergy. For the other tasks mentioned in the introduction, other specifications may
be required.
2 Normative references
Not applicable.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
accuracy
closeness of agreement between a measured quantity value and a true quantity value of a measurement
3.2
bench
long work table in a workshop or laboratory
3.3
clockwork
mechanism with a spring and toothed gearwheels, used to drive a mechanical clock, toy or other device
3.4
combined standard measurement uncertainty
obtained using the individual standard measurement uncertainties associated with the input quantities
in a measurement model
3.5
defatted
surface conditions after clearing with a fat removing substance
3.6
drum
cylindrical device for the mounting of a sticky tape
3.7
exine
outer wall of pollen grain, also called an exosporium
3.8
eyepiece
lens or combination of lenses in an optical instrument through which the eye views the image formed by
the objective lens or lenses; ocular
3.9
flow meter
instrument for measuring the flow rate of a fluid in a pipe
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3.10
flow rate
amount of fluid (e.g. air) that flows in a given time
3.11
fungal spore
reproductive unit of a fungi capable of giving rise to a new individual with or without sexual fusion
3.12
hood
metal cover or canopy for a stove, ventilator, etc.
3.13
impaction
sampling of airborne particles by inertial separation on any surface (e.g. of an adhesive)
3.14
magnetic stirrer
object or mechanical device used for stirring something
3.15
magnification
magnifying power of an instrument, e.g. this microscope should give a magnification of about × 100
3.16
microscope
optical instrument having a magnifying lens or a combination of lenses for inspecting objects too small to
be seen or too small to be seen distinctly and in detail by the unaided eye
3.17
objectives
optics (in a telescope, microscope, camera, or other optical system), the lens or combination of lenses,
that first receive the rays from the object and form the image in the focal plane of the eyepiece, as in a
microscope, or on a plate or screen as in a camera
Note 1 to entry: Also called object glass, object lens, objective lens.
3.18
orifice
opening or aperture, as of a tube or pipe; a mouthpiece with a slotlike opening on the side of the trap
3.19
particle
minute portion of matter
3.20
pollen
male gametophyte of flowering plants, consisting of microscopic grain discharged from the anthers
(Angiosperms) or from a male cone (Gymnosperms)
Note 1 to entry: Each pollen grain contains two male gametes (only one can fertilize the female ovule).
Note 2 to entry: Pollen are transported by wind, water, animals (e.g. insects).
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3.21
precision
closeness of agreement between indications or measured quantity values obtained by replicate
measurements on the same objects under specified conditions
3.22
repeatability
condition of measurement, out of a set of conditions that includes the same measurement procedure,
same operators, same measuring system, same operating conditions and same location, and replicate
measurements on the same objects over a short period of time
3.23
reproducibility
condition of measurement, out of a set of conditions that includes different locations, operators,
measuring systems, and replicate measurements on the same objects
3.24
sensitivity
in aerobiology, measurement of the proportion of search particle which is correctly identified
3.25
slide
rectangular piece of glass on which an object is mounted or placed for examination under a microscope
3.26
specificity
in aerobiology, measurement of the proportion of non-searched particles which are correctly identified
as different from the searched particles
3.27
standard measurement uncertainty
measurement uncertainty expressed as a standard deviation
3.28
suction
production of a negative pressure by the removal of air to force fluid into a vacant space
3.29
taxon/taxa
taxonomic group of any rank, such as a species, genus, family or other rank
3.30
trap
device used to collect something
3.31
vacuum
space from which the air has been completely or partly removed
3.32
vacuum pump
pump or device by which a partial vacuum can be produced
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3.33
wind vane
mechanical device attached to an elevated structure; rotates freely depending on the direction of the wind
4 Principle
Ambient air is sampled by a volumetric suction system and directed towards a suitably coated sampling
surface through a specific orifice oriented towards the wind; the particles contained in the sampled air
are deposited by impaction on a continuously moving adhesive acceptor surface. The sampling surface is
then examined with an optical microscope in order to identify and count the pollen and fungal spores per
area (deposition rates). Using this method allows to calculate concentrations as a daily mean or a 2-hour
mean. The sampling is usually done at low-volume rate (10 l/min). It allows a continuous sampling for
up to seven days [4] [5] [6].
5 Sampling
5.1 Equipment
5.1.1 Apparatus
5.1.1.1 Motorised suction pump
The motorised suction pump shall work 24 h a day and continuously throughout the year at the same
flow rate. The power supply may be either mains or battery driven (solar panels). The electric motor shall
be capable of continuous operation.
The suction system is, for instance, a vacuum pump. The flow rate of suction shall be adjusted by a flow
control valve.
The recommended flow rate is 10 l/min with an acceptable error in precision and accuracy of less than
10 % (± 1 l/min).
The validity of the calibration method recommended with an error of less than 10 % in the accuracy of
flow rate between calibration and operation shall be certified by the manufacturer. As the accuracy of the
calibration depends on the air resistance and characteristic curves of the individual trap, vacuum pump
and flow meter used [19], the validity has to be certified accordingly, i.e. specifically for the trap / vacuum
pump / flow meter combination delivered by the manufacturer.
The flow rate shall be checked at every change of the impaction support with a validated method.
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Key
1 lid
2 start reference pointer
3 lock nut
4 orifice start position
5 trapping surface
Figure 1a) — The Hirst volumetric trap showing 7-day lid assembly with drum
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Key
1 lid
3 lock nut
6 trapping surface on slide
Figure 1b) — The Hirst volumetric trap showing 24-h lid assembly with slide
5.1.1.2 Specific orifice
The orifice shall have the following dimensions (with associated tolerances):
— rectangular opening: 14 mm (±0,1 mm) × 2 mm (±0,1 mm);
— depth of the orifice: > 19 mm;
— distance from the inside orifice to the drum without the tape: 0,70 mm (±0,1 mm).
The depth allows the non-turbulence of laminar flow and directs the mixture of air and particles towards
the coated support. In consequence, an efficient particle impaction for pollen grains and fungal spores,
induced by the laminar flow, is ensured.
The outlet of the orifice shall be 0,70 mm (±0,1 mm) of the coated support (see Figure 2 – distance A-B).
The distance allows efficient particle impaction for pollen grains and fungal spores. It shall be controlled
[7] [8] [9].
The orifice should be directed into the air-stream using a wind vane.
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Key
1 drum
2 orifice
3 cover
A 20,5 mm or 22,5 mm, depending on supplier
B 19,8 mm or 21,8 mm, depending on supplier
C drum diameter 110 mm to 112 mm
A-B 0,7 ((± 0,1 mm)
Figure 2 — Pollen trap (Head of Hirst system)
5.1.2 Sampling support
5.1.2.1 General
Two possibilities are widely used depending on the requested sampling period as a sampling support:
— a glass slide for microscopy (76mm x 26mm) on which a transparent tape is fixed (48mm x 19mm)
coated with specific reagents (see Figure B.1 in Annex B) for one day of sampling.
— a drum (110 to 112mm in diameter) on which a transparent coated flexible tape is attached for
seven days of sampling. The length of this tape ranges from 345 to 350 mm (+/- 0,5mm) depending
on the size of the drum.
The sampling support is driven by a clockwork or an electric motor with a scrolling speed ranging from
2mm/h to 14mm/h (+-0,01mm/h) depending on the sampling period.
The sampling support shall scroll regularly in front of the back outlet of the orifice. Sampling shall always
be continuous and stable and not be stopped during the requested sampling period.
5.1.2.2 Transparent tape
The transparent tape is coated with an adhesive in order to fix the particles.
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The following requirements shall be fulfilled:
— The transparent tape shall be water-insoluble.
— The thickness of the whole transparent tape shall not be changed over time, and should not be
affected by operational conditions (temperature between – 20 °C to + 60 °C or humidity between 20
% and 100 %).
— It shall be perfectly transparent to allow the passing of microscopic light.
— The length shall be adapted to the support used.
5.1.2.3 Reagents
Two transparent coating products (solubilized in specific solvents or not) are useable: Vaseline
(petroleum jelly) or silicone.
5.1.2.3.1 Vaseline (petroleum jelly)
The main characteristics of Vaseline (petroleum jelly) (n° CAS: [8009 03 8], see Annex C, are the following:
— odorless;
— colorless;
— viscous liquid;
— boiling point: 68 °C;
— purity > 99 %;
— risk of diarrhea and other stomach problems if absorbed or in case of repeated exposure.
NOTE Material Safety Data Sheet (MSDS) need to be consulted to obtain special instructions before use (see
Annex C).
Solubilize the Vaseline (petroleum jelly) with toluene (n° CAS [108 88 3], see Annex C). It is also possible
to use insolubilized Vaseline (petroleum jelly) if there is a guarantee to have a regular thickness along the
strip.
The purity of Vaseline (petroleum jelly) and toluene shall be > 99 %.
5.1.2.3.2 Silicone
The main characteristics of silicone (n° CAS [90337 93 2], see Annex C) are the following:
— odorless;
— colorless to white;
— pasty;
— high viscosity stable from – 20 °C to +150 °C;
— flammable over 400 °C;
— irritating to eyes;
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NOTE Material Safety Data Sheet (MSDS) need to be consulted to obtain special instructions before use (see
Annex C).
— non-biodegradable.
Solubilize the silicone with a specific solvent (see Annex C).
The physical properties of the adhesive medium remain unaltered at temperatures between -20 °C and
+50 °C, making it suitable for the majority of bioclimatic zones.
5.1.3 Wind vane and rain shield
The wind vane allows permanent rotation of the trap head so that the orifice faces the wind. The rain
shield ensures a weather protection for the orifice (i.e. rainfall) (see Figure 3).
Key
1 wind vane
2 lid assembly
3 rain shield
4 orifice
5 rotation lock
6 motor cover
Figure 3a) — Schematic picture of a Hirst seven day volumetric spore sampling system
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Figure 3b) — Photo of a Hirst volumetric spore sampler [Source: RNSA]
5.1.4 Complete sampling system
The complete sampling system (so called “trap”) containing the motor, the vacuum pump, the orifice, the
rotating drum, the wind vane, the clockwork system, the impaction support shall be
— resistant to corrosion;
— well attached (i.e. resistant to wind-blow, etc.);
— always horizontal (at the head level).
The commercial devices that meet the requirements are presented in Annex A. For the different purposes,
refer to the specific publications.
For pollen grains and fungal spores monitoring purposes, the following requirements and conditions for
sampler positioning of the trap shall be fulfilled:
— The sampler shall be placed on a readily accessible, flat, horizontal surface. It should be on the roof
of a building, and away from the edge of the building in order to reduce the effects of turbulence.
— Care shall be taken to ensure that adjacent buildings do not screen the sampler or interfere with the
airflow. The sampler should be ideally placed on the roof of a building at more than 2 m from the
edge; the height above ground level depends on the city and on the height of neighbouring buildings.
— The sampler itself shall be elevated between 100 cm to 150 cm from the roof in order to avoid
turbulence between air layers and possible re-suspension of particles from the roof.
— The sampler shall not be placed in the vicinity of fixed or mobile sources of mass emission of
biological or non-biological particles. Proximity to non-biological particle sources may favour the
massive presence of residues in samples, which considerably hinder identification.
5.2 Operating procedure
5.2.1 Preparation of the coating medium
Prepare the coating medium as follows:
— Vaseline (petroleum jelly (18 g (± 1 g)) and toluene (1l) (purity > 99 %);
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— under the hood, add Vaseline (petroleum jelly) to toluene stirring until completely dispersed and
leave it to stand for about 48 h with periodic shaking to obtain a homogenous fluid solution;
— spread the solution on the tape, i.e. with a
...
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