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

Status
Published
Public Enquiry End Date
04-Dec-2017
Publication Date
25-Jun-2019
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
13-Jun-2019
Due Date
18-Aug-2019
Completion Date
26-Jun-2019

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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
...

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