SIST EN ISO 16014-5:2012

Plastics - Determination of average molecular mass and molecular mass distribution of polymers using size-exclusion chromatography - Part 5: Method using light-scattering detection (ISO 16014-5:2012)

This part of ISO 16014 specifies a general method for determining the average molecular mass and the molecular mass distribution of polymers using SEC-LS, i.e. size-exclusion chromatography coupled with lightscattering detection. The average molecular mass and the molecular mass distribution are calculated from molecular mass data and mass concentrations determined continuously with elution time. The molecular mass at each elution time is determined absolutely by combining a light-scattering detector with a concentrationsensitive detector. Therefore, SEC-LS is classified as an absolute method. For the applicability of the method, see ISO 16014-1:2012, Clause A.1.

Kunststoffe - Bestimmung der durchschnittlichen Molmasse und der Molmassenverteilung von Polymeren mittels Gelpermeationschromatographie - Teil 5: Lichtstreudetektionsverfahren (ISO 16014-5:2012)

Dieser Teil von ISO 16014 legt ein allgemeines Verfahren zum Bestimmen der durchschnittlichen Molmasse und der Molmassenverteilung von Polymeren durch GPC-LS, d. h. Gelpermeationschromatographie (GPC), gekoppelt mit Messung durch Lichtstreuung (LS), fest. Die durchschnittliche Molmasse und die Molmassenverteilung werden aus den Daten für die Molmasse und die Massenkonzentrationen berechnet, die kontinuierlich mit der Elutionszeit bestimmt werden. Die Molmasse bei jeder Elutionszeit wird als Absolutwert bestimmt, indem ein Detektor für die Lichtstreuung mit einem Konzentrationsdetektor kombiniert wird. Die GPC-LS wird deshalb als ein absolutes Verfahren klassifiziert.
Zur Anwendbarkeit des Verfahrens siehe ISO 16014-1:2012, Abschnitt A.1.

Plastiques - Détermination de la masse moléculaire moyenne et de la distribution des masses moléculaires de polymères par chromatographie d'exclusion stérique - Partie 5: Méthode utilisant la détection par diffusion lumineuse (ISO 16014-5:2012)

Polimerni materiali - Določevanje povprečne molekularne mase in porazdelitev molekularne mase polimerov z uporabo izključitvene kromatografije (SEC) - 5. del: Metoda s sipanjem svetlobe (ISO 16014-5:2012)

Ta del standarda ISO 16014 določa splošno metodo za ugotavljanje povprečne molekulske mase in porazdelitev molekulske mase polimerov z uporabo izključitvene kromatografije skupaj s sipanjem svetlobe (SEC-LS). Povprečna molekulska masa in porazdelitev molekulske mase se izračunata na podlagi podatkov o molekulski masi in koncentraciji mase, ki se kontinuirano določa z elucijskim časom. Molekulska masa se za posamezni elucijski čas določi absolutno s kombinacijo sipanja svetlobe in na koncentracijo občutljivega detektorja. Zato se izključitvena kromatografija skupaj s sipanjem svetlobe šteje za absolutno metodo. Za uporabo metode glejte točko A.1 standarda ISO 16014-1:2012.

General Information

• Status: Withdrawn
• Publication Date: 07-Aug-2012
• Withdrawal Date: 06-Aug-2019
• ICS: 83.080.01 - Plastics in general
• Technical Committee: IBLP - Paints and varnishes
• Current Stage: 9900 - Withdrawal (Adopted Project)
• Start Date: 24-Jul-2019
• Due Date: 16-Aug-2019
• Completion Date: 07-Aug-2019
• Ref Project: EN ISO 16014-5:2012 - Plastics - Determination of average molecular mass and molecular mass distribution of polymers using size-exclusion chromatography - Part 5: Method using light-scattering detection (ISO 16014-5:2012)

Relations

• Replaced By: SIST EN ISO 16014-5:2019 - Plastics - Determination of average molecular weight and molecular weight distribution of polymers using size-exclusion chromatography - Part 5: Light-scattering method (ISO 16014-5:2019)
• Effective Date: 01-Sep-2019

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 16014-5:2012
01-september-2012
3ROLPHUQLPDWHULDOL'RORþHYDQMHSRYSUHþQHPROHNXODUQHPDVHLQSRUD]GHOLWHY
PROHNXODUQHPDVHSROLPHURY]XSRUDERL]NOMXþLWYHQHNURPDWRJUDILMH6(&GHO
0HWRGDVVLSDQMHPVYHWOREH,62
Plastics - Determination of average molecular mass and molecular mass distribution of
polymers using size-exclusion chromatography - Part 5: Method using light-scattering
detection (ISO 16014-5:2012)
Kunststoffe - Bestimmung der durchschnittlichen Molmasse und der
Molmassenverteilung von Polymeren mittels Gelpermeationschromatographie - Teil 5:
Lichtstreudetektionsverfahren (ISO 16014-5:2012)
Plastiques - Détermination de la masse moléculaire moyenne et de la distribution des
masses moléculaires de polymères par chromatographie d'exclusion stérique - Partie 5:
Méthode utilisant la détection par diffusion lumineuse (ISO 16014-5:2012)
Ta slovenski standard je istoveten z: EN ISO 16014-5:2012
ICS:
83.080.01 Polimerni materiali na Plastics in general
splošno
SIST EN ISO 16014-5:2012 en,fr
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 16014-5:2012

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SIST EN ISO 16014-5:2012


EUROPEAN STANDARD
EN ISO 16014-5

NORME EUROPÉENNE

EUROPÄISCHE NORM
July 2012
ICS 83.080.01
English Version
Plastics - Determination of average molecular mass and
molecular mass distribution of polymers using size-exclusion
chromatography - Part 5: Method using light-scattering detection
(ISO 16014-5:2012)
Plastiques - Détermination de la masse moléculaire
Kunststoffe - Bestimmung der durchschnittlichen Molmasse
moyenne et de la distribution des masses moléculaires de und der Molmassenverteilung von Polymeren mittels
polymères par chromatographie d'exclusion stérique - Gelpermeationschromatographie - Teil 5:
Partie 5: Méthode utilisant la détection par diffusion Lichtstreudetektionsverfahren (ISO 16014-5:2012)
lumineuse (ISO 16014-5:2012)
This European Standard was approved by CEN on 30 June 2012.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16014-5:2012: E
worldwide for CEN national Members.

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SIST EN ISO 16014-5:2012
EN ISO 16014-5:2012 (E)
Contents Page
Foreword .3
2

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SIST EN ISO 16014-5:2012
EN ISO 16014-5:2012 (E)
Foreword
This document (EN ISO 16014-5:2012) has been prepared by Technical Committee ISO/TC 61 "Plastics" in
collaboration with Technical Committee CEN/TC 139 “Paints and varnishes” 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 January 2013, and conflicting national standards shall be withdrawn at
the latest by January 2013.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN/CENELEC Internal Regulations, the national standards organizations 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, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 16014-5:2012 has been approved by CEN as a EN ISO 16014-5:2012 without any
modification.

3

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SIST EN ISO 16014-5:2012

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SIST EN ISO 16014-5:2012
INTERNATIONAL ISO
STANDARD 16014-5
First edition
2012-07-01
Plastics — Determination of average
molecular mass and molecular mass
distribution of polymers using size-
exclusion chromatography —
Part 5:
Method using light-scattering detection
Plastiques — Détermination de la masse moléculaire moyenne
et de la distribution des masses moléculaires de polymères par
chromatographie d’exclusion stérique —
Partie 5: Méthode utilisant la détection par diffusion lumineuse
Reference number
ISO 16014-5:2012(E)
©
ISO 2012

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2012
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail [e-mail]
Web www.iso.org
Published in Switzerland
ii © ISO 2012 – All rights reserved

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
Contents Page
Foreword . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Symbols . 2
5 Principle . 2
5.1 SEC . 2
5.2 Light-scattering SEC . 2
6 Reagents . 2
6.1 Eluent . 2
6.2 Reagent for column evaluation . 3
6.3 Calibration standards . 3
6.4 Reagent for flow rate marker . 3
6.5 Additives . 3
7 Apparatus . 3
7.1 General . 3
7.2 Eluent reservoir . 4
7.3 Pumping system . 4
7.4 Injector . 4
7.5 Columns . 4
7.6 Detector . 5
7.7 Tubing . 5
7.8 Temperature control . 5
7.9 Recorder and plotter . 5
7.10 Data-processing system . 5
7.11 Other components . 5
8 Procedure . 5
8.1 Preparation of calibration solutions . 5
8.2 Preparation of a solution for determining the L-point . 5
8.3 Preparation of sample solutions . 5
8.4 Preparation of solutions for column performance evaluation . 6
8.5 Setting up the apparatus . 6
8.6 Operating parameters . 6
8.7 Number of determinations . 6
9 Calibration . 6
9.1 Calibration of concentration-sensitive detector and light-scattering detector . 6
9.2 Determination of delay volume . 8
9.3 Normalization of detector sensitivity . 8
9.4 Determination of refractive index increment . 8
10 Data acquisition and processing . 8
10.1 Data acquisition . 8
10.2 Evaluation of data and correction of chromatograms . 8
10.3 Data processing . 9
11 Expression of results .10
11.1 Calibration curve .10
11.2 Calculation of average molecular mass .12
11.3 Differential molecular mass distribution curve .12
11.4 Cumulative molecular mass distribution curve .12
12 Precision .13
13 Test report .13
© ISO 2012 – All rights reserved iii

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
13.1 General .13
13.2 Apparatus and measurement parameters .13
13.3 Calibration of the system .13
13.4 Calibration curve .14
13.5 Results .14
Annex A (informative) Round-robin test .15
Annex B (informative) Information on light scattering .17
Annex C (informative) Calibration curve in low molecular mass range .21
Bibliography .23
iv © ISO 2012 – All rights reserved

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 16014-5 was prepared by Technical Committee ISO/TC 61, Plastics, Subcommittee SC 5, Physical-
chemical properties.
ISO 16014 consists of the following parts, under the general title Plastics — Determination of average molecular
mass and molecular mass distribution of polymers using size-exclusion chromatography:
— Part 1: General principles
— Part 2: Universal calibration method
— Part 3: Low-temperature method
— Part 4: High-temperature method
— Part 5: Method using light-scattering detection
© ISO 2012 – All rights reserved v

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SIST EN ISO 16014-5:2012

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SIST EN ISO 16014-5:2012
INTERNATIONAL STANDARD ISO 16014-5:2012(E)
Plastics — Determination of average molecular mass and
molecular mass distribution of polymers using size-exclusion
chromatography —
Part 5:
Method using light-scattering detection
1 Scope
This part of ISO 16014 specifies a general method for determining the average molecular mass and the
molecular mass distribution of polymers using SEC-LS, i.e. size-exclusion chromatography coupled with light-
scattering detection. The average molecular mass and the molecular mass distribution are calculated from
molecular mass data and mass concentrations determined continuously with elution time. The molecular mass
at each elution time is determined absolutely by combining a light-scattering detector with a concentration-
sensitive detector. Therefore, SEC-LS is classified as an absolute method.
For the applicability of the method, see ISO 16014-1:2012, Clause A.1.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced document
(including any amendments) applies.
ISO 472, Plastics — Vocabulary
ISO 16014-1:2012, Plastics — Determination of average molecular mass and molecular mass distribution of
polymers using size-exclusion chromatography — Part 1: General principles
ISO 16014-2, Plastics — Determination of average molecular mass and molecular mass distribution of polymers
using size-exclusion chromatography — Part 2: Universal calibration method
ISO 16014-3:2012, Plastics — Determination of average molecular mass and molecular mass distribution of
polymers using size-exclusion chromatography — Part 3: Low-temperature method
ISO 16014-4:2012, Plastics — Determination of average molecular mass and molecular mass distribution of
polymers using size-exclusion chromatography — Part 4: High-temperature method
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 472 and ISO 16014-1 and the
following apply.
3.1
light-scattering detection
LS detection
a technique for determining the mass or size of polymer molecules in solution by measuring the light scattered
by the polymer molecules
© ISO 2012 – All rights reserved 1

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
3.2
refractive index increment
dn/dc
rate of change of the refractive index n of a polymer solution as a function of the mass concentration c
NOTE 1 It is also called the “specific refractive index increment” in the literature.
NOTE 2 The limiting value of dn/dc at zero concentration is commonly used in light scattering.
3.3
L-point
measured data point of a low molecular mass compound on the graph of molecular mass vs. elution time used
for justification of the polynominal fit of the calibration curve and/or construction of the calibration curve.
NOTE In the lower molecular mass region, the LS signal is too low to calculate molecular mass. Therefore the
measurement of the L-point is needed for justification or construction of the molecular mass calibration curve for the whole
range of molecular mass. The L-point is determined by measuring an oligomer of the polymer or an organic compound with
a similar chemical structure to the oligomer.
4 Symbols
R radius of gyration of a polymer molecule in solution nm
g
3 −2
A second virial coefficient for a polymer molecule in solution cm⋅mol⋅g
2
−3
c mass concentration of polymer in solution g⋅cm
−1
dn/dc refractive index increment cm⋅g
H excess signal intensity of a concentration detector at the ith elution time
i
I excess signal intensity of scattered light at the ith elution time
LS,i
3
V volume eluted during data acquisition time (interval) cm
e
5 Principle
5.1 SEC
For a discussion of size-exclusion chromatography in general, see ISO 16014-1:2012, Clause 4.
5.2 Light-scattering SEC
In SEC-LS, polymer molecules eluted from the SEC columns are irradiated by a beam of monochromatic
visible light. The light scattered by the molecules is continuously detected by a light-scattering detector. Since
the eluate is a dilute polymer solution, the intensity of the scattered light is approximately proportional to the
product of the molecular mass and the mass concentration of the polymer molecules. The scattered-light
intensity divided by the concentration therefore gives the molecular mass at a particular elution time. The
values of the molecular mass and the mass concentration or mass fraction at each elution time are used to
calculate the molecular mass distribution and the average molecular mass of the polymer.
6 Reagents
6.1 Eluent
For a general discussion of eluents, see ISO 16014-1:2012, 5.1.
For examples of eluents used for SEC measurements at temperatures below and above 60 °C, see Annex B of
ISO 16014-3:2012 and Annex B of ISO 16014-4:2012, respectively.
2 © ISO 2012 – All rights reserved

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
6.2 Reagent for column evaluation
For examples of low molecular mass compounds used for column evaluation, see ISO 16014-3:2012, 5.2, for
measurements at temperatures below 60 °C and ISO 16014-4:2012, 5.2, for those above 60 °C.
6.3 Calibration standards
Since the Rayleigh ratios of toluene and benzene are well-known, these solvents are recommended for
determining the calibration constant of the light-scattering detector (see Annex B, Clause B.2).
Aqueous solutions of potassium chloride (KCl) or sodium chloride (NaCl) are used for determining the calibration
constant of a refractive index detector. The concentration dependence of the differential refractive index of the
solutions is used to calculate the constant.
A low molecular mass, monodisperse polymer is used to determine the delay volume between the light-
scattering and concentration-sensitive detectors. This polymer may also be used to calibrate the angular
dependence of the detector sensitivity of a multiple-angle light-scattering detector. The radius of gyration R of
g
the polymer molecule used to calibrate the detector sensitivity, should preferably be less than 10 nm. A radius
of gyration less than 5 nm is desirable. Other compounds with a well-known R value may also be used.
g
Polymer reference materials are used for molecular mass calibration ranges from 20 000 to 50 000.
Low molecular mass organic compounds or oligomers of the polymer in the sample under investigation are
used for determining the “L-point”.
6.4 Reagent for flow rate marker
See ISO 16014-1:2012, 5.4.
For examples of compounds suitable for use as a flow rate marker, see ISO 16014-3:2012, 5.4, for measurements
at temperatures below 60 °C and ISO 16014-4:2012, 5.4, for those above 60 °C.
6.5 Additives
See ISO 16014-1:2012, 5.5.
Some examples of additives are given in ISO 16014-3:2012, 5.5, for measurements below 60 °C and
ISO 16014-4:2012, 5.5, for those above 60 °C.
7 Apparatus
7.1 General
A typical schematic diagram of an SEC-LS system is shown in Figure 1, which is similar to that shown in
ISO 16014-1:2012, Figure 1. The main difference is that a light-scattering detector is connected in series with
the concentration-sensitive detector. The light-scattering detector and concentration-sensitive detector may
also be connected in parallel. Any component that meets the performance requirements specified for this
method may be used.
Either commercially available SEC-LS systems or SEC-LS systems assembled in the laboratory may be used
for this method, provided they meet the levels of performance required.
© ISO 2012 – All rights reserved 3

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
Key
1 eluent reservoir 5 columns 9 computer
2 pump 6 light-scattering detector 10 printer
3 in-line filter 7 concentration-sensitive detector 11 to waste
4 injector 8 display
Figure 1 — Schematic diagram of a typical SEC-LS system
7.2 Eluent reservoir
See ISO 16014-1:2012, 6.2, and ISO 16014-3:2012, 6.2.
7.3 Pumping system
See ISO 16014-1:2012, 6.3, and ISO 16014-3:2012, 6.3.
7.4 Injector
See ISO 16014-1:2012, 6.4, and ISO 16014-3:2012, 6.4.
7.5 Columns
7.5.1 General
See ISO 16014-1:2012, 6.5.1, ISO 16014-3:2012, 6.5, and ISO 16014-4:2012, 6.5.
7.5.2 Determination of theoretical plate number
See ISO 16014-1:2012, 6.5.2.
7.5.3 Determination of resolution factor
See ISO 16014-1:2012, 6.5.3.
7.5.4 Determination of asymmetry factor
See ISO 16014-1:2012, 6.5.4.
4 © ISO 2012 – All rights reserved

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
7.6 Detector
7.6.1 Concentration-sensitive detector
See ISO 16014-1:2012, 6.6.
7.6.2 Light-scattering detector
This detector shall continuously monitor the intensity of the light scattered by the eluent coming off the columns.
Commercially available light-scattering detectors that may be used include single detectors set at a very low
angle and detectors which can be set up at two or more angles.
To avoid band-broadening of the chromatogram, the volume of the flow cell shall be as small as possible.
7.7 Tubing
See ISO 16014-1:2012, 6.7.
7.8 Temperature control
See ISO 16014-1:2012, 6.8.
7.9 Recorder and plotter
See ISO 16014-1:2012, 6.9.
7.10 Data-processing system
See ISO 16014-1:2012, 6.10.
7.11 Other components
See ISO 16014-1:2012, 6.11.
An in-line filter is necessary to remove any particulates which might cause noise (spikes) in the output of the
light-scattering detector.
8 Procedure
8.1 Preparation of calibration solutions
Prepare solutions of a monodisperse polymer for determining the delay volume between the two detectors.
The concentration of the solutions shall be such that the light-scattering detector and concentration-sensitive
detector provide a signal intensity sufficient for data handling. A typical concentration of the polymer is 5 mg/ml
to 10 mg/ml for low molecular mass polymers.
These polymer solutions may also be used for correcting or normalizing the sensitivity of the light-scattering detector.
8.2 Preparation of a solution for determining the L-point
A solution for determining the L-point may be prepared, if required, by dissolving appropriate oligomers or other low
molecular mass compounds in a suitable solvent. Typically, the concentration of this solution is 1 mg/ml to 5 mg/ml.
8.3 Preparation of sample solutions
See ISO 16014-3:2012, 7.2, for measurements below 60 °C and ISO 16014-4:2012, 7.2, for those above 60 °C.
© ISO 2012 – All rights reserved 5

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SIST EN ISO 16014-5:2012
ISO 16014-5:2012(E)
8.4 Preparation of solutions for column performance evaluation
See ISO 16014-3:2012, 7.3.
8.5 Setting up the apparatus
See ISO 16014-3:2012, 7.4.
8.6 Operating parameters
8.6.1 Flow rate
See ISO 16014-3:2012, 7.5.1.
8.6.2 Injection masses and injection volumes
See ISO 16014-3:2012, 7.5.2.
8.6.3 Column temperature
See ISO 16014-3:2012, 7.5.3.
8.6.4 Detector sensitivity
The signal intensity depends on the amount of sample injected, on the specific refractive index increment dn/dc
for a
...