SIST EN ISO 16373-3:2014

Textiles - Dyestuffs - Part 3: Method for determination of certain carcinogenic dyestuffs (method using triethylamine/methanol) (ISO 16373-3:2014)

EN ISO 16373-3 specifies a method for the detection and quantitative determination of the presence of carcinogenic dyestuffs as listed in Table 1 in dyed, printed or coated textile products by chromatographic analysis of their extracts.

Textilien - Farbstoffe - Teil 3: Verfahren zur Bestimmung von bestimmten karzinogenen Farbstoffen (Triethylamin/Methanol-Verfahren) (ISO 16373-3:2014)

Dieser Teil der ISO 16373 legt ein Verfahren zum Nachweis und zur quantitativen Bestimmung des Vorhandenseins der in Tabelle 1 aufgeführten karzinogenen Farbstoffe in gefärbten, bedruckten oder beschichteten Textilerzeugnissen durch eine chromatographische Analyse der Extrakte fest.

Textiles - Colorants - Partie 3: Méthode de détermination de certains colorants cancérogènes extractibles (méthode en utilisant la triéthylamine / méthanol) (ISO 16373-3:2014)

L'ISO 16373-3:2014 spécifie une méthode pour la détection et la détermination quantitative de la présence des colorants cancérigènes répertoriés dans les produits textiles teints, imprimés ou revêtus, par analyse chromatographique de leurs extraits.

Tekstilije - Barvila - 3. del: Metoda za ugotavljanje določenih kancerogenih barvil (metoda z uporabo trietilamina/metanola) (ISO 16373-3:2014)

Standard EN ISO 16373-3 določa metodo za odkrivanje in kvantitativno ugotavljanje prisotnosti določenih kancerogenih barvil, ki so navedena v tabeli 1, v barvanih, tiskanih ali prevlečenih tekstilnih izdelkih s pomočjo masne kromatografije njihovih ekstraktov.

General Information

• Status: Published
• Publication Date: 03-Jul-2014
• ICS: 59.080.01 - Textiles in general
• ICS: 87.060.10 - Pigments and extenders
• Technical Committee: ITEK - Textile and textile products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 05-Jun-2014
• Due Date: 10-Aug-2014
• Completion Date: 04-Jul-2014
• Ref Project: EN ISO 16373-3:2014 - Textiles - Dyestuffs - Part 3: Method for determination of certain carcinogenic dyestuffs (method using triethylamine/methanol) (ISO 16373-3:2014)

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 16373-3:2014
01-september-2014
7HNVWLOLMH%DUYLODGHO0HWRGD]DXJRWDYOMDQMHGRORþHQLKNDQFHURJHQLKEDUYLO
PHWRGD]XSRUDERWULHWLODPLQDPHWDQROD,62
Textiles - Dyestuffs - Part 3: Method for determination of certain carcinogenic dyestuffs
(method using triethylamine/methanol) (ISO 16373-3:2014)
Textilien - Farbstoffe - Teil 3: Verfahren zur Bestimmung von bestimmten karzinogenen
Farbstoffen (Triethylamin/Methanol-Verfahren) (ISO 16373-3:2014)
Textiles - Colorants - Partie 3: Méthode de détermination de certains colorants
cancérogènes extractibles (méthode en utilisant la triéthylamine / méthanol) (ISO 16373-
3:2014)
Ta slovenski standard je istoveten z: EN ISO 16373-3:2014
ICS:
59.080.01 Tekstilije na splošno Textiles in general
87.060.10 Pigmenti in polnila Pigments and extenders
SIST EN ISO 16373-3:2014 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 16373-3:2014

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SIST EN ISO 16373-3:2014

EUROPEAN STANDARD
EN ISO 16373-3

NORME EUROPÉENNE

EUROPÄISCHE NORM
June 2014
ICS 59.080.01
English Version
Textiles - Dyestuffs - Part 3: Method for determination of certain
carcinogenic dyestuffs (method using triethylamine/methanol)
(ISO 16373-3:2014)
Textiles - Colorants - Partie 3: Méthode de détermination de Textilien - Farbstoffe - Teil 3: Verfahren zur Bestimmung
certains colorants cancérigènes (méthode à la triéthylamine von bestimmten karzinogenen Farbstoffen
et au méthanol) (ISO 16373-3:2014) (Triethylamin/Methanol-Verfahren) (ISO 16373-3:2014)
This European Standard was approved by CEN on 17 April 2014.

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

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16373-3:2014 E
worldwide for CEN national Members.

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SIST EN ISO 16373-3:2014
EN ISO 16373-3:2014 (E)
Contents page
Foreword .3
2

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SIST EN ISO 16373-3:2014
EN ISO 16373-3:2014 (E)
Foreword
This document (EN ISO 16373-3:2014) has been prepared by Technical Committee ISO/TC 38 “Textiles” in
collaboration with Technical Committee CEN/TC 248 “Textiles and textile products” the secretariat of which is
held by BSI.
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 December 2014, and conflicting national standards shall be withdrawn
at the latest by December 2014.
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, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 16373-3:2014 has been approved by CEN as EN ISO 16373-3:2014 without any modification.


3

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SIST EN ISO 16373-3:2014

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SIST EN ISO 16373-3:2014
INTERNATIONAL ISO
STANDARD 16373-3
First edition
2014-06-01
Textiles — Dyestuffs —
Part 3:
Method for determination of certain
carcinogenic dyestuffs (method using
triethylamine/methanol)
Textiles - Colorants —
Partie 3: Méthode de détermination de certains colorants
cancérigènes (méthode à la triéthylamine et au méthanol)
Reference number
ISO 16373-3:2014(E)
©
ISO 2014

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2014
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested 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 2014 – All rights reserved

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Terms and definitions . 1
3 Principle . 1
4 Safety precautions . 2
4.1 General . 2
4.2 Handling. 2
5 Apparatus . 2
6 Reagents . 3
7 Test specimen sampling and preparation . 3
7.1 General . 3
8 Procedure. 3
8.1 Extraction . 3
8.2 Detection, identification and quantification of carcinogenic dyestuffs . 4
9 Test report . 4
Annex A (informative) Chromatographic analysis . 5
Annex B (informative) Round robin test results .22
Bibliography .27
© ISO 2014 – All rights reserved iii

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2. www.iso.org/directives
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received. www.iso.org/patents
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity
assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers
to Trade (TBT) see the following URL: Foreword - Supplementary information
The committee responsible for this document is ISO/TC 38.
ISO 16373 consists of the following parts, under the general title Textiles — Dyestuffs:
— Part 1: General principles of testing coloured textiles for dyestuff identification
— Part 2: General method for the determination of extractable dyestuffs including allergenic and
carcinogenic dyestuffs (method using pyridine-water)
— Part 3: Method for determination of certain carcinogenic dyestuffs (method using triethylamine/methanol)
iv © ISO 2014 – All rights reserved

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

Introduction
Due to concerns of consumers over safety and hygiene, many countries have introduced regulations
regarding carcinogenic dyestuffs in textile articles. To support international and national regulations
the development of a test method is very important and this part of ISO 16373 does just that.
The ISO 16373 series deal with dyestuffs used in textile for qualification and quantification.
1)
— ISO 16373-1 includes the definition of the dyestuff, and classes the description of some procedures
to identify qualitatively the dyestuff class used in textile material. The other parts of ISO 16373 are
related to the quantification of some dyestuffs.
— In ISO 16373-2, the principle of the test method is based on extraction using pyridine-water solution,
which has been found to be the most efficient solution to extract a large range of dyestuffs, including
allergenic and carcinogenic dyestuffs.
— In this part of ISO 16373, the principle of the test method is based on extraction using triethylamine-
methanol solution. This solution has been found to be efficient at extracting some dyestuffs in some
cases.
Additional information related to the recovery rate (to characterize the extraction efficiency) obtained
from the application of ISO 16373-2 and this part of ISO 16373 is summarized in ISO 16373-1:—, Annex B.
It is important to note that there are other test methods related to azo dyes, for which a reduction of the
extracted azo dyes leads to the release of some aromatic amines to be detected and determined using
[6][7]
chromatography.
1) To be published.
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SIST EN ISO 16373-3:2014
INTERNATIONAL STANDARD ISO 16373-3:2014(E)
Textiles — Dyestuffs —
Part 3:
Method for determination of certain carcinogenic dyestuffs
(method using triethylamine/methanol)
1 Scope
This part of ISO 16373 specifies a method for the detection and quantitative determination of the
presence of carcinogenic dyestuffs as listed in Table 1 in dyed, printed or coated textile products by
chromatographic analysis of their extracts.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1
textile
woven fabric, knitted fabric, etc., formed by the interlocking of fibres and yarns having a certain cohesion
and which is generally intended for clothing or furniture applications
Note 1 to entry: Textiles often include certain types of non-woven fabrics.
2.2
carcinogenic dyestuff
substance yielding a dye that is a substance known to be or suspected of being a human carcinogen
3 Principle
The dyestuff of a coloured test specimen is extracted by means of a solvent in an ultrasonic bath under
specified conditions. The extract is analysed using either a high-performance liquid chromatography
photodiode array detector (HPLC-DAD) or a high-performance liquid chromatography mass spectrometer
(HPLC-MSD).
The carcinogenic dyestuffs are listed in Table 1.
Table 1 — List of carcinogenic dyestuffs
C.I. Constitution
C.I. Generic name CAS number
number
   C.I. Basic Red 9   569–61–9 42500
   C.I. Disperse Orange 11   82–28–0 60700
   C.I. Disperse Yellow 3   2832–40–8 11855
   C.I. Acid Red 114   6459–94–5 23635
   C.I. Acid Red 26   3761–53–3 16150
   C.I. Direct Black 38   1937–37–7 30235
   C.I. Direct Red 28   573–58–0 22120
   C.I. Disperse Blue 1   2475–45–8 64500
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

Table 1 (continued)
C.I. Constitution
C.I. Generic name CAS number
number
   C.I. Basic Violet 14   632–99–5 42510
   C.I. Direct Blue 6   2602–46–2 22610
   C.I. Direct Brown 95   16071–86–6 30145
4 Safety precautions
4.1 General
Warning — The dyestuffs targeted in this part of ISO 16373 are classified as substances known
to be or suspected of being human carcinogens.
4.2 Handling
It is the user’s responsibility to ensure any handling and disposal of these substances are in strict
accordance with the appropriate national health and safety regulations.
It is the user’s responsibility to use safe and proper techniques in handling materials in this test
method. Consult manufacturers for specific details, such as material safety data sheets and other
recommendations.
Good laboratory practice should be followed. Wear safety glasses in all laboratory areas and
single-use dust respirator while handling the dyestuff powder.
5 Apparatus
5.1 Ultrasonic bath, capable of heating to and maintaining at (50 ± 5) °C and output power of 40 W,
oscillating frequency, 42 kHz, or equivalent.
5.2 Coil condenser, for chemical testing use.
5.3 Vacuum rotary evaporator, capable of operating at water evaporation capacity of a maximum of
25 ml/min, or equivalent.
5.4 Round bottom flask, of 200 ml in capacity.
5.5 Pipettes, of 1 ml and 10 ml in capacity.
5.6 Volumetric flask, of 10 ml, 100 ml and 1 l in capacity.
5.7 High-performance liquid chromatography (HPLC) system and diode array detector (DAD) or
mass spectroscope (MSD).
5.8 Test tube, of 100 ml in capacity, with a silicone plug.
NOTE For details of the high-performance liquid chromatography equipment, see Annex A.
5.9 Analytical balance, of 0,001 g in resolution.
2 © ISO 2014 – All rights reserved

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

6 Reagents
Unless otherwise specified, use only reagents of recognized analytical grade.
6.1 Acetonitrile.
6.2 Methanol.
6.3 Hexane.
6.4 0,25 % tri-ethylamine methanol solution, 2,5 ml triethylamine is dissolved in methanol and
made up to 1 l.
6.5 10 mmol/l ammonium acetate aqueous solution, 0,77 g ammonium acetate is dissolved in water
and made up to 1 l.
6.6 Carcinogenic dyestuffs. Use only carcinogenic dyestuffs of reagent grade of the highest purity
available on the market, or dyestuffs of which quantities of the dye are manufactured in controlled
environments within Europe under the control of the EU creating standard dyestuffs.
6.7 Standard solution of carcinogenic dyestuffs.
An amount of each carcinogenic dyestuff is weighed accurately in the range of 1 mg to 10 mg and
transferred quantitatively to a 10 ml volumetric flask, and then made up to volume with methanol (6.2)
to prepare a standard solution in the range of 100 ug/ml to 1 000 ug/ml.
The standard solution may be diluted properly and four solutions with known concentrations may be
made to draw the calibration curve. As an example, the range of concentration of standard solutions for
the calibration curve can be recommended to be from 1 μg/ml to 100 μg/ml.
7 Test specimen sampling and preparation
7.1 General
The test specimen shall be selected based on the following criteria:
— parts of the textile article;
— nature of the fibre component (fibre composition);
— colours.
Prepare a test specimen of maximum 1,0 g (≤1,0 g) by cutting the laboratory sample up into small pieces
2
no larger than 1 cm . Determine the mass of the test specimen to the nearest 0,01 g and record it as m
E
(see 8.2).
8 Procedure
8.1 Extraction
8.1.1 Cleansing
If required, remove oil, grease or other fatty matter from the surface of the test specimen by soaking it
in 100 ml hexane (6.3) for 5 min in an ultrasonic bath (5.1) at ambient temperature.
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

Remove and drain the test specimen.
8.1.2 Extraction of dyestuff
Place 1,0 g of the test specimen in a 100 ml test tube. Add 100 ml of the 0,25 % tri-ethylamine methanol
solution (6.4) and seal the test tube using a silicone plug. Heat the tube in an ultrasonic bath until a
temperature of 50 °C ± 2 °C is reached and maintained this temperature for 3 h.
8.1.3 Concentration of extract and preparation of analysis solution
Transfer the extract obtained according to 8.1.2 to a 200 ml round bottom flask (5.4) and place it in a
vacuum rotary evaporator (5.3) in the water bath at 40 °C ± 2 °C until all the liquid has evaporated.
Dissolve the residue in 1 ml of methanol. Filter the solution through a 0,45 μm PTFE filter. If the resultant
measurement is higher than the calibrated range of the chromatograph, dilute the solution further with
methanol.
8.2 Detection, identification and quantification of carcinogenic dyestuffs
Detection of carcinogenic dyestuffs is performed by the chromatographic analysis using the apparatus
described in 5.7. When the carcinogenic dyestuffs are identified by comparing with peaks of reference
carcinogenic dyestuffs, quantification is performed using a calibration curve, which is drawn by using a
minimum of four points obtained from an HPLC analysis of the standard solution (6.7) and the correlation
coefficient of the linear curve should be 0,99 in the range of concentration of 1 μg/ml to 100 μg/ml.
Quantification is executed by the method of HPLC/DAD. When a large amount of foreign substances are
detected, HPLC/MSD is recommended for identification and quantification.
The concentration of carcinogenic dyestuff in the specimen is calculated as a mass fraction of the
specimen, w (μg/g), as given by Formula (1):
ρ ×V
s
w = (1)
m
E
where
ρ is the interpolated concentration of carcinogenic dyestuff, in micrograms per millilitre (μg/
s
ml);
V is the final solution volume made up to according to 8.1.2, in millilitres (ml);
m is the mass of the test specimen, in millilitres (ml);
E
9 Test report
The test report shall include the following:
a) reference to this part of ISO 16373, i.e. ISO 16373-3;
b) kind, origin and designation of the specimen (partial specimen, if applicable);
c) detection method and quantification method;
d) results reported as level and detection limit for each of the carcinogenic dyestuffs (μg/g);
e) any deviation from the procedure.
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

Annex A
(informative)

Chromatographic analysis
A.1 Chromatographic analysis — General
As the instrumental equipment of laboratories might vary, no generally applicable instructions can
be provided for chromatograph analysis. Therefore, the following parameters have been successfully
tested and used. See Figures A.1 to A.14 and Table A.1.
A.2 High-performance liquid chromatography/diode array detector (HPLC/DAD)
See Table A.1.
Table A.1 — Condition of HPLC/DAD
Eluent 1: 10 mmol/l ammonium acetate
Eluent 2: Acetonitrile
Column Inertsil ODS-3, 150 mm × 3,0 mm, 5 μm
Flow rate: 0,8ml/min
Gradient Time (min) Eluent 2 concentrations
Time programme Initial       5 %
30         60 %
40         60 %
40,1         5 %
50           5 %
Column temperature: 45 °C
Injection volume: 5,0 μl
Determination: DAD
Quantification: 540 nm (for Basic Red 9)
480 nm (for Disperse Orange 11)
350 nm (for Disperse Yellow 3)
510 nm (for Acid Red 114)
510 nm (for Acid Red 26)
600 nm (for Direct Black 38)
500 nm (for Direct Red 28)
Remark   Columns of equivalent quality may be used.
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

mAU
542,14
20
15
10
286,87
5
0
200 300 400 500 600 700
nm
Figure A.1 — UV spectrum of Basic Red 9
mAU
30
20
Basic Red 9
10
0
05 10 15 20 25 30 35 40
min
Figure A.2 — HPLC/DAD Chromatogram at 540 nm detection
mAU
247,83
20
15
10
477,32
5
0
200 300 400 500 600 700
nm
Figure A.3 — UV spectrum of Disperse Orange 11
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

mAU
10,0
7,5
5,0
Disperse Orange 11
2,5
0,0
05 10 15 20 25 30 35 40
min
Figure A.4 — HPLC/DAD Chromatogram at 480 nm detection
mAU
9
354,40
8
7
6
249,50
5
4
3
2
1
0
200 300 400 500 600 700
nm
Figure A.5 — UV spectrum of Disperse Yellow 3
mAU
10,0
7,5
Disperse Yellow 3
5,0
2,5
0,0
05 10 15 20 25 30 35 40
min
Figure A.6 — HPLC/DAD Chromatogram at 350 nm detection
mAU
35
30
25
511,92
20
15
364,27
10
5
0
200 300 400 500 600 700
nm
Figure A.7 — UV spectrum of Acid Red 114
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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

mAU
15
10
Acid Red 114
5
0
05 10 15 20 25 30 35 40
min
Figure A.8 — HPLC/DAD Chromatogram at 510 nm detection
mAU
30
507,93
25
20
15
327,64
10
5
0
200 300 400 500 600 700
nm
Figure A.9 — UV spectrum of Acid Red 26
mAU
15
10
Acid Red 26
5
0
05 10 15 20 25 30 35 40
min
Figure A.10 — HPLC/DAD Chromatogram at 510 nm detection
mAU
9
601,17
8
7 470,13
6
5
4
3
2
1
0
300 400 500 600 700
nm
Figure A.11 — UV spectrum of Direct Black 38
8 © ISO 2014 – All rights reserved

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

mAU
10,5
7,5
Direct Black 38
5,0
2,5
0,0
05 10 15 20 25 30 35 40
min
Figure A.12 — HPLC/DAD Chromatogram at 600 nm detection
mAU
40
503,73
35
339,23
30
25
20
15
10
5
0
300 400 500 600 700
nm
Figure A.13 — UV spectrum of Direct Red 28
mAU
40
30
Direct Red 28
20
10
0
05 10 15 20 25 30 35 40
min
Figure A.14 — HPLC/DAD Chromatogram at 500 nm detection
A.3 High-performance liquid chromatography/mass analysis detector
(HPLC/MSD)
A.3.1 LC/MS SIM (selected ion monitoring) method
See Figures A.15 to A.28 and Table A.2.
Table A.2 — Condition of the HPLC/MSD
Eluent 1 10 mmol/l ammonium acetate
Eluent 2 Acetonitrile
Column Inertsil ODS-3, 150 mm × 3,0 mm, 5 μm
Flow rate 0,8 ml/min
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ISO 16373-3:2014(E)

Table A.2 (continued)
Gradient Time (min) Eluent 2 concentrations
Time programme Initial       5 %
30         60 %
40         60 %
40,1        5 %
50          5 %
Column temperature: 45 °C
Injection volume: 5,0 μl
Detection: Four pile pole or ion trap mass detector
SIM(selected ion monitoring) method
Mass spectrum
Ionization: ESI electro spray ionizing method and positive/negative
ion detection
Monitor channel: positive Q1 m/z 288 (for Basic Red 9)
positive Q1 m/z 238 (for Disperse Orange 11)
positive Q1 m/z 270 (for Disperse Yellow 3)
negative Q1 m/z 785 (for Acid Red 114)
negative Q1 m/z 435 (for Acid Red 26)
positive Q1 m/z 738 (for Direct Black 38)
positive Q1 m/z 653 (for Direct Red 28)
Impressed voltage: 5 000 V
Temperature of spray: 500 °C
Remark   Columns of equivalent quality may be used.
288
6,0e7
Basic Red 9
5,0e7 m/z 288
4,0e7
3,0e7
2,0e7
1,0e7
5,0e6
200 300 400 500 600 700 800 900 1000 1100 1200
m/z, amu
Figure A.15 — Mass spectrum of Basic Red 9
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18,1
9,0e5
8,0e5
7,0e5
6,0e5
5,0e5
Basic Red 9
4,0e5
3,0e5
2,0e5
1,0e5
5,0e4
0.0
24 68 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50
Time, min
Figure A.16 — SIM Chromatogram of Basic Red 9
238
2,0e7
Disperse Orange 11
1,8e7
m/z 238
1,6e7
1,4e7
1,2e7
1,0e7
8,0e6
6,0e6
4,0e6
2,0e6
200 300 400 500 600 700 800 900 1000 1100 1200
m/z, amu
Figure A.17 — Mass spectrum of Disperse Orange 11
2,8e5
25,9
2,4e5
2,0e5
Disperse Orange 11
1,6e5
1,2e5
8,0e4
4,0e4
26 10 14 18 22 26 30 34 38 42 46 50
Time, min
Figure A.18 — SIM chromatogram of Disperse Orange 11
© ISO 2014 – All rights reserved 11

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SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

270
3,0e7
Disperse Yellow 3
2,6e7
m/z 270
2,2e7
1,8e7
1,4e7
1,0e7
6,0e6
2,0e6
200 300 400 500 600 700 800 900 1000 1100 1200
m/z, amu
Figure A.19 — Mass spectrum of Disperse Yellow 3
26,4
8,0e4
7,0e4
6,0e4 Disperse Yellow 3
5,0e4
4,0e4
3,0e4
2,0e4
1,0e4
26 10 14 18 22 26 30 34 38 42 46 50
Time, min
Figure A.20 — SIM chromatogram of Disperse Yellow 3
1,8e7
Acid Red 114
1,6e7
m/z 785
1,4e7
1,2e7
1,0e7
8,0e6
6,0e6
4,0e6
2,0e6
m/z, amu
Figure A.21 — Mass spectrum of Acid Red 114
12 © ISO 2014 – All rights reserved
785
392
422
200 300 400 500 600 700 800 900 1000 1100 1200

---------------------- Page: 24 ----------------------

SIST EN ISO 16373-3:2014
ISO 16373-3:2014(E)

2,0e4
1,8e4
1,6e4
Acid Red 114
1,4e4
1,2e4
1,0e4
8000
6000
4000
2000
Time, min
Figure A.22 — SIM chromatogram of Acid Red 114
1,8e7
Acid Red 26
1,6e7
m/z 435
1,4e7
1,2e7
1,0e7
8,0e6
6,0e6
4,0e6
2,0e6
m/z, amu
Figure A.23 — Mass spectrum of Acid Red 26
11,2
1,6e5
1,4e5
Acid Red 26
1,2e5
1,0e5
8,0e4
6,0e4
4,0e4
2,0e4
26 10 14 18 22 26 30 34 38 42 46 50
Time, min
Figure A.24 — SIM chromatogram of Acid Red 2
...