SIST EN 71-11:2006
(Main)Safety of toys - Part 11: Organic chemical compounds - Methods of analysis
Safety of toys - Part 11: Organic chemical compounds - Methods of analysis
This Part 11 of the European Standard EN 71 for safety of toys specifies methods for the analysis of toy and toy material extracts prepared according to the sampling procedures in EN 71-10, to enable assessment of compliance with the chemical requirements specified in EN 71-9.
This European Standard specifies analytical methods for the identification and determination of the following groups of organic chemicals:
Flame retardants
Colourants
Primary aromatic amines
Monomers and solvents
Wood preservatives
Preservatives
Plasticisers
NOTE 1 Methods for formaldehyde in accessible textile components of toys; accessible paper components of toys; and accessible resin-bonded wood components of toys are specified in EN 71-9.
NOTE 2 The method for free formaldehyde as a preservative is specified in EN 71-10.
Sicherheit von Spielzeug - Teil 11: Organisch-chemische Verbindungen, Analysenverfahren
Dieser Teil 11 der Europäischen Norm EN 71 zur Sicherheit von Spielzeug legt Verfahren für die Analyse von Spielzeug- und Spielzeugmaterialextrakten fest, die nach den Probenahmeverfahren in EN 71-10 hergestellt wurden, um die Bewertung der Übereinstimmung mit den in EN 71-9 festgelegten chemischen Anforderungen zu ermöglichen.
Diese Norm legt Analysenverfahren zur qualitativen und quantitativen Bestimmung der folgenden Gruppen organischer Chemikalien fest:
- Flammschutzmittel (Flammverzögerungsmittel);
- Farbstoffe;
- primäre aromatische Amine;
- nicht-flüchtige Monomere und Lösemittel;
- flüchtige Monomere und Lösemittel;
- Holzschutzmittel;
- Konservierungsstoffe;
- Weichmacher.
ANMERKUNG 1 Verfahren für die qualitative und quantitative Bestimmung von Formaldehyd in zugänglichen textilen Bestandteilen von Spielzeug, zugänglichen Bestandteilen aus Papier und harzgebundenen Holzteilen von Spielzeug sind in EN 71-9 festgelegt.
ANMERKUNG 2 Das Verfahren für die Bestimmung von freiem Formaldehyd als Konservierungsstoff ist in EN 71-10 festgelegt.
Sécurité des jouets - Partie 11 : Composés organiques chimiques - Méthodes d'analyse
La présente partie 11 de la Norme européenne EN 71 relative a la sécurité des jouets spécifie des méthodes d’analyse des extraits de jouets et de matériaux jouets préparés selon les procédures d’échantillonnage de l’EN 71-10 pour permettre d’évaluer la conformité avec les exigences chimiques spécifiées dans l’EN 71-9.
La présente Norme européenne spécifie des méthodes d’analyse destinées a l’identification et au dosage des groupes suivants de produits chimiques organiques :
?retardateurs de flamme ;
?colorants ;
?amines aromatiques primaires ;
?monomeres et solvants ;
?produits de préservation du bois ;
?agents de conservation ;
?plastifiants.
NOTE 1 Les méthodes applicables au formaldéhyde dans les parties textile, papier, en bois stratifié avec résine, accessibles de jouets, sont spécifiées dans l'EN 71-9.
NOTE 2 La méthode applicable au formaldéhyde libre en tant que conservateur est spécifiée dans l'EN 71-10.
Varnost igrač – 11. del: Organske kemijske spojine – Analizne metode
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 71-11:2006
01-februar-2006
Varnost igrač – 11. del: Organske kemijske spojine – Analizne metode
Safety of toys - Part 11: Organic chemical compounds - Methods of analysis
Sicherheit von Spielzeug - Teil 11: Organisch-chemische Verbindungen,
Analysenverfahren
Sécurité des jouets - Partie 11 : Composés organiques chimiques - Méthodes d'analyse
Ta slovenski standard je istoveten z: EN 71-11:2005
ICS:
97.200.50 Igrače Toys
SIST EN 71-11:2006 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 71-11:2006
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SIST EN 71-11:2006
EUROPEAN STANDARD
EN 71-11
NORME EUROPÉENNE
EUROPÄISCHE NORM
November 2005
ICS 97.200.50
English Version
Safety of toys - Part 11: Organic chemical compounds - Methods
of analysis
Sécurité des jouets - Partie 11 : Composés chimiques Sicherheit von Spielzeug - Teil 11: Organisch-chemische
organiques dans les jouets - Méthodes d'analyse Verbindungen - Analysenverfahren
This European Standard was approved by CEN on 27 June 2005.
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 Central Secretariat 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 Central Secretariat has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia,
Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2005 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 71-11:2005: E
worldwide for CEN national Members.
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SIST EN 71-11:2006
EN 71-11:2005 (E)
Contents
Page
Foreword .3
Introduction.4
1 Scope .5
2 Normative references .5
3 Terms and definitions.5
4 Environmental, health and safety precautions .6
5 Methods of analysis.6
5.1 General .6
5.2 Flame retardants .6
5.3 Colourants.9
5.4 Primary aromatic amines.15
5.5 Monomers and solvents .19
5.6 Wood preservatives.37
5.7 Preservatives.41
5.8 Plasticisers.44
Annex A (informative) Methods of analysis for volatile solvents .48
Annex B (informative) Validation of test methods .65
Annex C (informative) Colourants – conformational analysis .66
Annex ZA (informative) Clauses of this European Standard addressing essential requirements or
other provisions of EU Directives.68
Bibliography.69
2
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SIST EN 71-11:2006
EN 71-11:2005 (E)
Foreword
This European Standard (EN 71-11:2005) has been prepared by Technical Committee CEN/TC 52 “Safety of
Toys”, the secretariat of which is held by DS.
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 May 2006, and conflicting national standards shall be withdrawn at the
latest by May 2006.
This European Standard has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association, and supports essential requirements of EU Directive(s).
For relationship with EU Directive(s), see informative Annex ZA, which is an integral part of this European
Standard.
This European Standard constitutes part 11 of the European Standard on Safety of Toys.
This part should be read in conjunction with parts 9 and 10.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic,
Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland
and United Kingdom.
3
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SIST EN 71-11:2006
EN 71-11:2005 (E)
Introduction
The European Standard EN 71 for “safety of toys” consists of the following parts:
Part 1: Mechanical and physical properties
Part 2: Flammability
Part 3: Migration of certain elements
Part 4: Experimental sets for chemistry and related activities
Part 5: Chemical toys (sets) other than experimental sets
Part 6: Graphical symbols for age warning labelling
Part 7: Finger paints – Requirements and test methods
Part 8: Swings, slides and similar activity toys for indoor and outdoor family domestic use
Part 9: Organic chemical compounds – Requirements
Part 10: Organic chemical compounds – Sample preparation and extraction
Part 11: Organic chemical compounds – Methods of analysis
The European Standards EN 71-9, EN 71-10 and EN 71-11 were mandated by the European Commission
(M/229) to address the risks presented by organic chemical compounds in toys by taking into account the
potential exposure and toxicological effects of those substances considered to present the greatest risk to
health.
This European Standard specifies methods of analysis to enable assessment of compliance with the chemical
requirements specified in EN 71-9 when toy and toy material extracts have been prepared according to the
sampling procedures in EN 71-10.
This part on methods of analysis should be read in conjunction with EN 71-9, which contains requirements for
certain organic chemical compounds in toys, and EN 71-10, which describes sample preparation and
extraction procedures.
This European Standard takes into account the opinion of the Toxicology Section of the Scientific Advisory
Committee published in 1992 (EUR 13976), which recommended that certain groups of chemical compounds
used in toys and toy materials need to be given special attention. In drafting this European Standard CEN/TC
52 has considered organic chemicals that can be classified within the following groups:
Solvents
Preservatives
1
Plasticisers (excluding phthalate plasticisers)
Flame retardants
Monomers
Biocides (wood preservatives)
Processing aids
Colouring agents
1
Phthalate plasticisers were specifically excluded from the scope of mandate M/229.
4
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SIST EN 71-11:2006
EN 71-11:2005 (E)
1 Scope
This Part 11 of the European Standard EN 71 for safety of toys specifies methods for the analysis of toy and
toy material extracts prepared according to the sampling procedures in EN 71-10, to enable assessment of
compliance with the chemical requirements specified in EN 71-9.
This European Standard specifies analytical methods for the identification and determination of the following
groups of organic chemicals:
Flame retardants
Colourants
Primary aromatic amines
Monomers and solvents
Wood preservatives
Preservatives
Plasticisers
NOTE 1 Methods for formaldehyde in accessible textile components of toys; accessible paper components of toys; and
accessible resin-bonded wood components of toys are specified in EN 71-9.
NOTE 2 The method for free formaldehyde as a preservative is specified in EN 71-10.
2 Normative references
The following referenced documents are indispensable for the application of this European Standard. For
dated references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
EN 71-9:2005, Safety of toys – Part 9: Organic chemical compounds – Requirements
EN 71-10:2005, Safety of toys – Part 10: Organic chemical compounds – Sample preparation and extraction
EN ISO 3696, Water for analytical laboratory use – Specification and test methods (ISO 3696:1987)
3 Terms and definitions
For the purposes of this European Standard, the following terms and definitions apply.
3.1
action limit
routinely-achievable limit of quantification for a particular substance using the specified method of analysis
3.2
aqueous migrate
liquid obtained after extracting a toy material according to the procedure specified in Clause 6 of EN 71-
10:2005
3.3
test portion
portion of the laboratory sample prepared for analysis
3.4
toy material
material from which toys and toy components are made
NOTE This definition differs from that given in EN 71-3
5
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EN 71-11:2005 (E)
4 Environmental, health and safety precautions
When preparing this European Standard, consideration was given to the minimisation of environmental
impacts caused by the use of the methods of analysis.
It is the users’ responsibility to use safe and proper techniques in handling materials in the methods of
analysis specified in this European Standard.
Consult manufacturers for specific details such as material safety data sheets and other
recommendations.
Wear protective goggles and coats in all laboratory areas.
Be careful about substances, which are toxic and/or human carcinogens.
A fume cupboard shall be used during preparation of organic solvent solutions.
Solvents shall be disposed of in accordance with environmental requirements.
5 Methods of analysis
5.1 General
All chemicals used for analysis shall be of analytical grade (pro analysis) or, if unavailable, the best technical
grade. Water shall be of grade 3 according to EN ISO 3696 or of a comparable quality, and demonstrably free
from analytes of interest.
The precision of volumetric glassware should be grade A.
The analysis of toys and toy materials for chemical compounds for which limits are given in Tables 2 A to 2 I
of EN 71-9:2005 shall be performed in accordance with the sampling procedures specified in EN 71-10 and
the methods of analysis described in this European Standard. Alternative methods of analysis are acceptable
only if they are capable of achieving at least the accuracy and precision of the methods described in this
European Standard; an adequate sensitivity; and have been validated to show that the results are equivalent
to those of these standard methods.
5.2 Flame retardants
NOTE Methods are given for pentabromodiphenyl ether and octabromodiphenyl ether in order to enable compliance
with Directive 2003/11/EC of the European Parliament and of the Council to be demonstrated for textile toy materials.
5.2.1 Principle
Flame retardants are determined in acetonitrile extracts of toy materials by liquid chromatography with diode-
array and mass spectrometry detection (LC-DAD-MS) using the external standard method of calibration.
6
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EN 71-11:2005 (E)
5.2.2 Standards, reagents and solvents
5.2.2.1 Standards
2*
5.2.2.1.1 Pentabromodiphenyl ether , CAS No. 32534-81-9
3
5.2.2.1.2 Octabromodiphenyl ether * , CAS No. 32536-52-0
5.2.2.1.3 Tri-o-cresyl phosphate, CAS No. 78-30-8
5.2.2.1.4 Tris(2-chloroethyl) phosphate, CAS No. 115-96-8
5.2.2.2 Reagents and solvents
5.2.2.2.1 Acetonitrile
5.2.2.2.2 Dichloromethane
5.2.2.2.3 Ammonium acetate, anhydrous
5.2.2.2.4 Acetic acid, glacial
5.2.2.2.5 Ammonium acetate, 10 mmol/l aqueous solution, pH 3,6
Transfer (0,77 0,01) g ammonium acetate (5.2.2.2.3) into a 1 000-ml volumetric flask, add 980 ml of water,
±
adjust the pH to 3,6 ± 0,1 with glacial acetic acid and make up to the mark with water.
5.2.2.3 Stock standard solution (100 mg/l)
Weigh, to the nearest 0,1 mg, (10 ± 1) mg of each flame retardant (5.2.2.1) into a 100-ml volumetric flask.
Add 25 ml of acetonitrile (5.2.2.2.1) and mix carefully to dissolve. Place in an ultrasonic bath for 10 min to
ensure complete dissolution. Make up to the mark with acetonitrile.
The stability of the mixed stock standard solution should be checked regularly. It should be stable for up to 6
months when stored in the dark at (4 ± 2) °C.
5.2.3 Apparatus
Liquid chromatograph with diode-array and mass spectrometer detectors
The following LC-DAD-MS conditions for flame retardant determination have been found to be suitable:
4
Column: C18, 80 Å, 3,5 µm, double endcapped, (Zorbax Eclipse XDB , or equivalent)
2,1 mm x 150 mm
Guard column: C18, 80 Å, 4 mm x 2,0 mm,
Mobile phase A: Ammonium acetate solution, 10 mmol/l, pH 3,6 (5.2.2.2.5)
Mobile phase B: Acetonitrile
Gradient: see Table 1
2
This substance is also known as pentabromodiphenyl oxide.
*
There are no requirements in EN 71-9 for this substance.
3
This substance is also known as octabromodiphenyl oxide.
4
Zorbax Eclipse XDB is an example of a suitable product available commercially. This information is given for the
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
7
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EN 71-11:2005 (E)
Injection volume: 5 µl
Run time: 45 min
Flow: 0,3 ml/min
DAD mode: 240 nm ± 20 nm
DAD range: 200 nm to 800 nm
Nebulizer: 200 Kpa
Dry gas: 10 l/min
MS range: 110 m/z to 500 m/z
MS mode: Scan positive
Ionisation: ESI+
Fragmentor: 80 V
Table 1 – Gradient program
Time Mobile phase A Mobile phase B
min % %
0 60 40
7 40 60
17 2 98
35 2 98
45 60 40
5.2.4 Procedure
5.2.4.1 Calibration solutions
Prepare a series of mixed flame retardant calibration solutions from the stock standard solution (5.2.2.3) at
1,0 mg/l, 2,0 mg/l, 4,0 mg/l and 8,0 mg/l concentrations in acetonitrile.
5.2.4.2 Determination
Proceed to liquid chromatographic determination using the conditions described in 5.2.3. Inject the calibration
solutions (5.2.4.1) and the extract obtained at 8.1.1 of EN 71-10:2005.
5.2.4.3 Identification
For a positive identification, the peak purity factor should achieve a match of at least 85 %.
5.2.5 Calculation of analyte concentration
Determine the concentration of a flame retardant in the acetonitrile extract from a calibration graph produced
from the calibration solutions.
Calculate the concentration of a flame retardant in the sample using the following equation:
C [mg /l ]
comp, solvent
Conc [mg / kg ] = x 10 (1)
A
where
C is the concentration of a flame retardant in acetonitrile extract
comp, solvent
A is the mass in grams of the test portion taken for analysis (see 8.1.1 of EN 71-10:2005).
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5.2.6 Limits and precision
Table 2 – Limits and precision
Component Action limit RSD Recovery
mg/kg % at 5 mg/l % at 100 mg/kg
(equivalent to from fabric
50 mg/kg in sample)
a
Pentabromodiphenyl ether 2,0 103
(total of 3 isomers)
a
Octabromodiphenyl ether 1,2 99
(total of 4 isomers)
Tri-o-cresyl phosphate 50 2,4 69
Tris(2-chloroethyl) 50 2,6 102
phosphate
a
The limit in Directive 2003/11/EC is 0,1 % by mass (1 000 mg/kg)
Correlation coefficient (r): > 0,995
5.2.7 Test report
The test report shall contain the following information:
a) description and identification of the product and material tested;
b) reference to this European Standard;
c) identification of flame retardants in the extract of the test portion;
d) amount of each flame retardant identified expressed as a concentration (mg/kg) in the toy material;
e) any deviations from the test procedure specified;
f) date of test.
5.3 Colourants
5.3.1 Principle
Colourants are identified and semi-quantified in extracts of toy materials by liquid chromatography with diode-
array detection (LC-DAD). If a positive identification is obtained, confirmation can be achieved using liquid
chromatography with mass spectrometry detection (LC-MS).
5.3.2 Standards, reagents and solvents
NOTE Pure materials for these colourants are not readily available and their composition can vary. A supplier of a
suitable colourant is indicated for each analyte.
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5
5.3.2.1 Standards
5.3.2.1.1 Disperse Blue 1, C.I. 64500
e.g. Sigma Aldrich 21 564-3
5.3.2.1.2 Disperse Blue 3, C.I. 61505
e.g. Sigma Aldrich 21 565-1
5.3.2.1.3 Disperse Blue 106
e.g. Fluka 28241
5.3.2.1.4 Disperse Blue 124
e.g. Fluka 21620
5.3.2.1.5 Disperse Yellow 3, C.I. 11855
e.g. Sigma Aldrich 21 568-6
5.3.2.1.6 Disperse Orange 3, C.I. 11005
e.g. Sigma Aldrich 36 479-7
5.3.2.1.7 Disperse Orange 37
e.g. Fluka 21603
5.3.2.1.8 Disperse Red 1, C.I. 11110
e.g. Sigma Aldrich 34 420-6
5.3.2.1.9 Solvent Yellow 1, C.I. 11000
e.g. Sigma Aldrich 18 636-8
5.3.2.1.10 Solvent Yellow 2, C.I. 11020
e.g. Sigma Aldrich 11 449-9
5.3.2.1.11 Solvent Yellow 3, C.I. 11160
e.g. Sigma Aldrich 12 156-8
5.3.2.1.12 Basic Red 9, C.I. 42500
e.g. Sigma Aldrich 21 559-7
5
The suppliers of the colourants mentioned in this subclause are examples of suppliers of suitable products available
commercially. This information is given for the convenience of users of this European Standard and does not constitute an
endorsement by CEN.
10
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5.3.2.1.13 Basic Violet 1, C.I. 42535
e.g. Sigma Aldrich 19 809-9
5.3.2.1.14 Basic Violet 3, C.I.42555
e.g. Sigma Aldrich 86 099-9
5.3.2.1.15 Acid Red 26, C.I.16150
e.g. Sigma Aldrich 19 976-1
5.3.2.1.16 Acid Violet 49, C.I. 42640
e.g. Sigma Aldrich S334294
5.3.2.2 Reagents and solvents
5.3.2.2.1 Tetrabutylammonium hydroxide solution, 40 % in water
5.3.2.2.2 Citric acid
5.3.2.2.3 Ammonium acetate, anhydrous
5.3.2.2.4 Acetonitrile
5.3.2.2.5 Tetrahydrofuran
5.3.2.2.6 Ethanol, absolute
5.3.2.2.7 Ammonium hydroxide, approx. 35 % (V/V)
5.3.2.2.8 Acetic acid, glacial
5.3.2.2.9 Ammonium acetate, 10 mmol/l aqueous solution, pH 3,6
Transfer (0,77 ± 0,01) g ammonium acetate (5.3.2.2.3) into a 1 000-ml volumetric flask, add 980 ml of water,
adjust the pH to 3,6 ± 0,1 with glacial acetic acid and make up to the mark with water.
5.3.2.2.10 Citrate-buffered tetrabutylammonium hydroxide solution
Transfer (13,6 ± 0,1) g tetrabutylammonium hydroxide solution (5.3.2.2.1) and (2,8 ± 0,1) g citric acid into a
1 000-ml volumetric flask, add 980 ml of water, adjust the pH to 9,0 ± 0,1 with ammonium hydroxide
(5.3.2.2.7) and make up to the mark with water.
5.3.3 Standard solutions
5.3.3.1 General
When preparing stock solutions of each colourant, purity values shall be taken into account. Store the stock
standard solutions in a refrigerator at (4 ± 2) °C.
11
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5.3.3.2 Stock standard solution (50 µg/ml), mix 1
Weigh, to the nearest 0,1 mg, (5 ± 1) mg of each colourant listed below into a 100-ml volumetric flask. Add
50 ml of ethanol (5.3.2.2.6) and mix carefully to dissolve. Place in an ultrasonic bath for 15 min to ensure
complete dissolution. Make up to the mark with ethanol.
Disperse Blue 1
Disperse Blue 106
Disperse Blue 124
Disperse Orange 3
Disperse Orange 37
Solvent Yellow 1
Solvent Yellow 2
Solvent Yellow 3
Basic Red 9
Basic Violet 1
Basic Violet 3
5.3.3.3 Stock standard solution (50 µg/ml), mix 2
Weigh, to the nearest 0,1 mg, (5 ± 1) mg of each colourant listed below into a 100-ml volumetric flask. Add
50 ml of ethanol and mix carefully to dissolve. Place in an ultrasonic bath for 15 min to ensure complete
dissolution. Make up to the mark with ethanol.
Disperse Blue 3
Disperse Yellow 3
Disperse Red 1
Acid Red 26
Acid Red 49
5.3.4 Apparatus
5.3.4.1 PTFE membrane filter, 0,45 µm
5.3.4.2 Ultrasonic bath
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5.3.4.3 Liquid chromatograph with diode-array detector
The following LC-DAD conditions for colourant analysis have been found to be suitable:
6
Column: C18, 100 Å, 5 µm, endcapped, (Luna C18(2) , or equivalent), 250 mm x 4,6 mm
6
Guard column: 2 x C18, 100 Å, 5 µm, endcapped, (Luna C18(2) , or equivalent)
Column temperature: 25 °C
Mobile phase A: Citrate-buffered tetrabutylammonium hydroxide solution (5.3.2.2.10)
Mobile phase B: Tetrahydrofuran
Mobile phase C: Acetonitrile
Gradient: see Table 3
Run time: 45 min
Flow rate 0,8 ml/min
Injection volume: 5 µl to 50 µl
Analysis time: 35 min
Wavelength range: 275 nm to 760 nm
Resolution factor: 4,8 nm
Acquisition rate: 1 spectrum/second
Table 3 – Gradient program
Time Mobile phase A Mobile phase B Mobile phase C
min % % %
0 80,0 10,0 10,0
2,50 80,0 10,0 10,0
30,0 5,0 48,0 47,0
35,0 5,0 48,0 47,0
45,0 80,0 10,0 10,0
5.3.5 Procedure
5.3.5.1 Calibration solutions
Prepare two series of colourant calibration solutions from the stock standard solutions of mix 1 (5.3.3.2) and
mix 2 (5.3.3.3) at 1 mg/l, 2 mg/l, 3 mg/l, 4 mg/l and 5 mg/l concentrations in ethanol.
5.3.5.2 Determination
Proceed to liquid chromatographic determination using the conditions described in 5.3.4.3. Inject the
calibration solutions of both mix 1 and mix 2 (5.3.5.1) and the ethanol phase obtained at 8.1.3, 8.2.1, 8.3.1,
8.4.1, 8.5.1, 8.6.1, 8.7.1, 8.8.1 or 8.9.1 of EN 71-10:2005, as appropriate.
5.3.5.3 Identification
For a positive identification, the peak purity factor should achieve a match of at least 85 %.
6
Luna C18(2) is an example of a suitable product available commercially. This information is given for the convenience of
users of this European Standard and does not constitute an endorsement by CEN of this product.
13
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5.3.6 Calculation of analyte concentration
Determine the concentration of a colourant in the ethanolic extract from a calibration graph produced from the
calibration solutions.
Calculate the concentration of a colourant in the sample using the following equation:
C [mg /l ]
comp, solvent
Conc [mg / kg ] = xD (2)
A
where
C is the concentration of a colourant in ethanolic extract
comp, solvent
A is the mass in grams of the test portion taken for analysis (see 8.1.3, 8.2.1, 8.3.1, 8.4.1,
8.5.1, 8.6.1, 8.7.1, 8.8.1 or 8.9.1 of EN 71-10:2005, as appropriate)
D is the dilution factor; generally 10, but see 8.5 of EN 71-10:2005 for aqueous liquid toy
material samples.
5.3.7 Limits and precision
Table 4 – Limits and precision
Substance Action limit RSD
mg/kg % at 5 mg/l
(equivalent to 10 mg/kg
in a sample)
Disperse Blue 1 10 1,8
Disperse Blue 3 10 4,9
Disperse Blue 106 10 4,4
Disperse Blue 124 10 2,2
Disperse Yellow 3 10 0,3
Disperse Orange 3 10 1,6
Disperse Orange 37 10 2,8
Disperse Red 1 10 1,6
Solvent Yellow 1 10 1,1
Solvent Yellow 2 10 1,1
Solvent Yellow 3 10 1,6
Basic Red 9 10 1,1
Basic Violet 1 10 1,5
Basic Violet 3 10 1,0
Acid Red 26 10 2,1
Acid Violet 49 10 1,4
Correlation coefficient (r): > 0,995
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5.3.8 Additional information
5.3.8.1 LC-DAD spectral library
It is advisable to compile a spectral library of all colours listed in 5.3.2.1 using the processing software that is
used to operate the liquid chromatograph. Details of retention times and Lambda max of each colourant
should be recorded together with peak purity data, if available.
It has been observed that several of the colourants separated into two or more chromatographic peaks. The
colourants concerned were C.I. Acid Red 26, C.I. Disperse Blue 3, C.I. Acid Violet 49 and C.I. Basic Violet 1.
In an attempt to characterise these peaks, an LC-MS method was developed in tandem with the LC-DAD
method. The conditions used for the LC-MS analysis are detailed in Annex C.
5.3.9 Test report
The test report shall contain the following information:
a) description and identification of the product and material tested;
b) reference to this European Standard;
c) identification of colourants in the ethanolic extract of the test portion;
d) amount of each colourant identified expressed as a concentration (mg/kg) in the toy material;
e) whether a confirmation test has been carried out and, if so, the technique used and its result;
f) any deviations from the test procedure specified;
g) date of test.
5.4 Primary aromatic amines
5.4.1 Principle
Aromatic amines are determined in extracts of toy materials by gas chromatography with mass spectrometry
detection (GC-MS) using the external standard method of calibration combined with appropriate internal
standards.
15
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SIST EN 71-11:2006
EN 71-11:2005 (E)
5.4.2 Standards, reagents and solvents
5.4.2.1 Standards
5.4.2.1.1 Benzidine
5.4.2.1.2 Aniline
5.4.2.1.3 2-Naphthylamine
5.4.2.1.4 3,3’-Dichlorobenzidine
5.4.2.1.5 3,3’-Dimethoxybenzidine
5.4.2.1.6 3,3’-Dimethylbenzidine
5.4.2.1.7 2-Methoxyaniline
5.4.2.1.8 o-Toluidine
5.4.2.1.9 4-Chloroaniline
5.4.2.2 Reagents and solvents
Acetonitrile
5.4.2.2.1
tert-Butyl methyl ether
5.4.2.2.2
5.4.2.2.3 n-Hexane
7
5.4.2.2.4 Chromabond XTR (Porous granulated kieselguhr)
5.4.2.3 Stock standard solution (100 mg/l)
Weigh, to the nearest 0,1 mg, (10 ± 1) mg of each aromatic amine (5.4.2.1) into a 100-ml volumetric flask.
Add 25 ml of acetonitrile (5.4.2.2.1) and mix carefully to dissolve. Place in an ultrasonic bath for 10 min to
ensure complete dissolution. Make up to the mark with acetonitrile.
The stability of the mixed stock standard solution should be checked regularly. It should be stable for up to 6
months when stored in the dark at (4 ± 2) °C.
7
Chromabond XTR is an example of a suitable product available commercially. This information is given for the
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
16
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EN 71-11:2005 (E)
5.4.3 Apparatus
5.4.3.1 Ultrasonic bath
5.4.3.2 Vortex® shaker
5.4.3.3 Centrifuge
5.4.3.4 Gas chromatograph with mass spectrometer detector
Ensure that the gas chromatograph has been fully cleaned before analysis, as the determination of aromatic
amines by this technique is affected by other contaminants. It is recommended that the inlet sleeve is
deactivated and an amine-specific column used.
The following GC-MS conditions for primary aromatic amine determination have been found to be suitable:
Injector:
Mode: Splitless 0,5 min
Carrier gas: Helium 0,8 ml/min
I
...
SLOVENSKI STANDARD
SIST EN 71-11:2006
01-februar-2006
9DUQRVWLJUDþ±GHO 2UJDQVNHNHPLMVNHVSRMLQH±$QDOL]QHPHWRGH
Safety of toys - Part 11: Organic chemical compounds - Methods of analysis
Sicherheit von Spielzeug - Teil 11: Organisch-chemische Verbindungen,
Analysenverfahren
Sécurité des jouets - Partie 11 : Composés organiques chimiques - Méthodes d'analyse
Ta slovenski standard je istoveten z: EN 71-11:2005
ICS:
97.200.50 ,JUDþH Toys
SIST EN 71-11:2006 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 71-11:2006
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SIST EN 71-11:2006
EUROPEAN STANDARD
EN 71-11
NORME EUROPÉENNE
EUROPÄISCHE NORM
November 2005
ICS 97.200.50
English Version
Safety of toys - Part 11: Organic chemical compounds - Methods
of analysis
Sécurité des jouets - Partie 11 : Composés chimiques Sicherheit von Spielzeug - Teil 11: Organisch-chemische
organiques dans les jouets - Méthodes d'analyse Verbindungen - Analysenverfahren
This European Standard was approved by CEN on 27 June 2005.
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 Central Secretariat 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 Central Secretariat has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia,
Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2005 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 71-11:2005: E
worldwide for CEN national Members.
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SIST EN 71-11:2006
EN 71-11:2005 (E)
Contents
Page
Foreword .3
Introduction.4
1 Scope .5
2 Normative references .5
3 Terms and definitions.5
4 Environmental, health and safety precautions .6
5 Methods of analysis.6
5.1 General .6
5.2 Flame retardants .6
5.3 Colourants.9
5.4 Primary aromatic amines.15
5.5 Monomers and solvents .19
5.6 Wood preservatives.37
5.7 Preservatives.41
5.8 Plasticisers.44
Annex A (informative) Methods of analysis for volatile solvents .48
Annex B (informative) Validation of test methods .65
Annex C (informative) Colourants – conformational analysis .66
Annex ZA (informative) Clauses of this European Standard addressing essential requirements or
other provisions of EU Directives.68
Bibliography.69
2
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EN 71-11:2005 (E)
Foreword
This European Standard (EN 71-11:2005) has been prepared by Technical Committee CEN/TC 52 “Safety of
Toys”, the secretariat of which is held by DS.
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 May 2006, and conflicting national standards shall be withdrawn at the
latest by May 2006.
This European Standard has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association, and supports essential requirements of EU Directive(s).
For relationship with EU Directive(s), see informative Annex ZA, which is an integral part of this European
Standard.
This European Standard constitutes part 11 of the European Standard on Safety of Toys.
This part should be read in conjunction with parts 9 and 10.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic,
Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland
and United Kingdom.
3
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SIST EN 71-11:2006
EN 71-11:2005 (E)
Introduction
The European Standard EN 71 for “safety of toys” consists of the following parts:
Part 1: Mechanical and physical properties
Part 2: Flammability
Part 3: Migration of certain elements
Part 4: Experimental sets for chemistry and related activities
Part 5: Chemical toys (sets) other than experimental sets
Part 6: Graphical symbols for age warning labelling
Part 7: Finger paints – Requirements and test methods
Part 8: Swings, slides and similar activity toys for indoor and outdoor family domestic use
Part 9: Organic chemical compounds – Requirements
Part 10: Organic chemical compounds – Sample preparation and extraction
Part 11: Organic chemical compounds – Methods of analysis
The European Standards EN 71-9, EN 71-10 and EN 71-11 were mandated by the European Commission
(M/229) to address the risks presented by organic chemical compounds in toys by taking into account the
potential exposure and toxicological effects of those substances considered to present the greatest risk to
health.
This European Standard specifies methods of analysis to enable assessment of compliance with the chemical
requirements specified in EN 71-9 when toy and toy material extracts have been prepared according to the
sampling procedures in EN 71-10.
This part on methods of analysis should be read in conjunction with EN 71-9, which contains requirements for
certain organic chemical compounds in toys, and EN 71-10, which describes sample preparation and
extraction procedures.
This European Standard takes into account the opinion of the Toxicology Section of the Scientific Advisory
Committee published in 1992 (EUR 13976), which recommended that certain groups of chemical compounds
used in toys and toy materials need to be given special attention. In drafting this European Standard CEN/TC
52 has considered organic chemicals that can be classified within the following groups:
Solvents
Preservatives
1
Plasticisers (excluding phthalate plasticisers)
Flame retardants
Monomers
Biocides (wood preservatives)
Processing aids
Colouring agents
1
Phthalate plasticisers were specifically excluded from the scope of mandate M/229.
4
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SIST EN 71-11:2006
EN 71-11:2005 (E)
1 Scope
This Part 11 of the European Standard EN 71 for safety of toys specifies methods for the analysis of toy and
toy material extracts prepared according to the sampling procedures in EN 71-10, to enable assessment of
compliance with the chemical requirements specified in EN 71-9.
This European Standard specifies analytical methods for the identification and determination of the following
groups of organic chemicals:
Flame retardants
Colourants
Primary aromatic amines
Monomers and solvents
Wood preservatives
Preservatives
Plasticisers
NOTE 1 Methods for formaldehyde in accessible textile components of toys; accessible paper components of toys; and
accessible resin-bonded wood components of toys are specified in EN 71-9.
NOTE 2 The method for free formaldehyde as a preservative is specified in EN 71-10.
2 Normative references
The following referenced documents are indispensable for the application of this European Standard. For
dated references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
EN 71-9:2005, Safety of toys – Part 9: Organic chemical compounds – Requirements
EN 71-10:2005, Safety of toys – Part 10: Organic chemical compounds – Sample preparation and extraction
EN ISO 3696, Water for analytical laboratory use – Specification and test methods (ISO 3696:1987)
3 Terms and definitions
For the purposes of this European Standard, the following terms and definitions apply.
3.1
action limit
routinely-achievable limit of quantification for a particular substance using the specified method of analysis
3.2
aqueous migrate
liquid obtained after extracting a toy material according to the procedure specified in Clause 6 of EN 71-
10:2005
3.3
test portion
portion of the laboratory sample prepared for analysis
3.4
toy material
material from which toys and toy components are made
NOTE This definition differs from that given in EN 71-3
5
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SIST EN 71-11:2006
EN 71-11:2005 (E)
4 Environmental, health and safety precautions
When preparing this European Standard, consideration was given to the minimisation of environmental
impacts caused by the use of the methods of analysis.
It is the users’ responsibility to use safe and proper techniques in handling materials in the methods of
analysis specified in this European Standard.
Consult manufacturers for specific details such as material safety data sheets and other
recommendations.
Wear protective goggles and coats in all laboratory areas.
Be careful about substances, which are toxic and/or human carcinogens.
A fume cupboard shall be used during preparation of organic solvent solutions.
Solvents shall be disposed of in accordance with environmental requirements.
5 Methods of analysis
5.1 General
All chemicals used for analysis shall be of analytical grade (pro analysis) or, if unavailable, the best technical
grade. Water shall be of grade 3 according to EN ISO 3696 or of a comparable quality, and demonstrably free
from analytes of interest.
The precision of volumetric glassware should be grade A.
The analysis of toys and toy materials for chemical compounds for which limits are given in Tables 2 A to 2 I
of EN 71-9:2005 shall be performed in accordance with the sampling procedures specified in EN 71-10 and
the methods of analysis described in this European Standard. Alternative methods of analysis are acceptable
only if they are capable of achieving at least the accuracy and precision of the methods described in this
European Standard; an adequate sensitivity; and have been validated to show that the results are equivalent
to those of these standard methods.
5.2 Flame retardants
NOTE Methods are given for pentabromodiphenyl ether and octabromodiphenyl ether in order to enable compliance
with Directive 2003/11/EC of the European Parliament and of the Council to be demonstrated for textile toy materials.
5.2.1 Principle
Flame retardants are determined in acetonitrile extracts of toy materials by liquid chromatography with diode-
array and mass spectrometry detection (LC-DAD-MS) using the external standard method of calibration.
6
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EN 71-11:2005 (E)
5.2.2 Standards, reagents and solvents
5.2.2.1 Standards
2*
5.2.2.1.1 Pentabromodiphenyl ether , CAS No. 32534-81-9
3
5.2.2.1.2 Octabromodiphenyl ether * , CAS No. 32536-52-0
5.2.2.1.3 Tri-o-cresyl phosphate, CAS No. 78-30-8
5.2.2.1.4 Tris(2-chloroethyl) phosphate, CAS No. 115-96-8
5.2.2.2 Reagents and solvents
5.2.2.2.1 Acetonitrile
5.2.2.2.2 Dichloromethane
5.2.2.2.3 Ammonium acetate, anhydrous
5.2.2.2.4 Acetic acid, glacial
5.2.2.2.5 Ammonium acetate, 10 mmol/l aqueous solution, pH 3,6
Transfer (0,77 0,01) g ammonium acetate (5.2.2.2.3) into a 1 000-ml volumetric flask, add 980 ml of water,
±
adjust the pH to 3,6 ± 0,1 with glacial acetic acid and make up to the mark with water.
5.2.2.3 Stock standard solution (100 mg/l)
Weigh, to the nearest 0,1 mg, (10 ± 1) mg of each flame retardant (5.2.2.1) into a 100-ml volumetric flask.
Add 25 ml of acetonitrile (5.2.2.2.1) and mix carefully to dissolve. Place in an ultrasonic bath for 10 min to
ensure complete dissolution. Make up to the mark with acetonitrile.
The stability of the mixed stock standard solution should be checked regularly. It should be stable for up to 6
months when stored in the dark at (4 ± 2) °C.
5.2.3 Apparatus
Liquid chromatograph with diode-array and mass spectrometer detectors
The following LC-DAD-MS conditions for flame retardant determination have been found to be suitable:
4
Column: C18, 80 Å, 3,5 µm, double endcapped, (Zorbax Eclipse XDB , or equivalent)
2,1 mm x 150 mm
Guard column: C18, 80 Å, 4 mm x 2,0 mm,
Mobile phase A: Ammonium acetate solution, 10 mmol/l, pH 3,6 (5.2.2.2.5)
Mobile phase B: Acetonitrile
Gradient: see Table 1
2
This substance is also known as pentabromodiphenyl oxide.
*
There are no requirements in EN 71-9 for this substance.
3
This substance is also known as octabromodiphenyl oxide.
4
Zorbax Eclipse XDB is an example of a suitable product available commercially. This information is given for the
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
7
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EN 71-11:2005 (E)
Injection volume: 5 µl
Run time: 45 min
Flow: 0,3 ml/min
DAD mode: 240 nm ± 20 nm
DAD range: 200 nm to 800 nm
Nebulizer: 200 Kpa
Dry gas: 10 l/min
MS range: 110 m/z to 500 m/z
MS mode: Scan positive
Ionisation: ESI+
Fragmentor: 80 V
Table 1 – Gradient program
Time Mobile phase A Mobile phase B
min % %
0 60 40
7 40 60
17 2 98
35 2 98
45 60 40
5.2.4 Procedure
5.2.4.1 Calibration solutions
Prepare a series of mixed flame retardant calibration solutions from the stock standard solution (5.2.2.3) at
1,0 mg/l, 2,0 mg/l, 4,0 mg/l and 8,0 mg/l concentrations in acetonitrile.
5.2.4.2 Determination
Proceed to liquid chromatographic determination using the conditions described in 5.2.3. Inject the calibration
solutions (5.2.4.1) and the extract obtained at 8.1.1 of EN 71-10:2005.
5.2.4.3 Identification
For a positive identification, the peak purity factor should achieve a match of at least 85 %.
5.2.5 Calculation of analyte concentration
Determine the concentration of a flame retardant in the acetonitrile extract from a calibration graph produced
from the calibration solutions.
Calculate the concentration of a flame retardant in the sample using the following equation:
C [mg /l ]
comp, solvent
Conc [mg / kg ] = x 10 (1)
A
where
C is the concentration of a flame retardant in acetonitrile extract
comp, solvent
A is the mass in grams of the test portion taken for analysis (see 8.1.1 of EN 71-10:2005).
8
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EN 71-11:2005 (E)
5.2.6 Limits and precision
Table 2 – Limits and precision
Component Action limit RSD Recovery
mg/kg % at 5 mg/l % at 100 mg/kg
(equivalent to from fabric
50 mg/kg in sample)
a
Pentabromodiphenyl ether 2,0 103
(total of 3 isomers)
a
Octabromodiphenyl ether 1,2 99
(total of 4 isomers)
Tri-o-cresyl phosphate 50 2,4 69
Tris(2-chloroethyl) 50 2,6 102
phosphate
a
The limit in Directive 2003/11/EC is 0,1 % by mass (1 000 mg/kg)
Correlation coefficient (r): > 0,995
5.2.7 Test report
The test report shall contain the following information:
a) description and identification of the product and material tested;
b) reference to this European Standard;
c) identification of flame retardants in the extract of the test portion;
d) amount of each flame retardant identified expressed as a concentration (mg/kg) in the toy material;
e) any deviations from the test procedure specified;
f) date of test.
5.3 Colourants
5.3.1 Principle
Colourants are identified and semi-quantified in extracts of toy materials by liquid chromatography with diode-
array detection (LC-DAD). If a positive identification is obtained, confirmation can be achieved using liquid
chromatography with mass spectrometry detection (LC-MS).
5.3.2 Standards, reagents and solvents
NOTE Pure materials for these colourants are not readily available and their composition can vary. A supplier of a
suitable colourant is indicated for each analyte.
9
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EN 71-11:2005 (E)
5
5.3.2.1 Standards
5.3.2.1.1 Disperse Blue 1, C.I. 64500
e.g. Sigma Aldrich 21 564-3
5.3.2.1.2 Disperse Blue 3, C.I. 61505
e.g. Sigma Aldrich 21 565-1
5.3.2.1.3 Disperse Blue 106
e.g. Fluka 28241
5.3.2.1.4 Disperse Blue 124
e.g. Fluka 21620
5.3.2.1.5 Disperse Yellow 3, C.I. 11855
e.g. Sigma Aldrich 21 568-6
5.3.2.1.6 Disperse Orange 3, C.I. 11005
e.g. Sigma Aldrich 36 479-7
5.3.2.1.7 Disperse Orange 37
e.g. Fluka 21603
5.3.2.1.8 Disperse Red 1, C.I. 11110
e.g. Sigma Aldrich 34 420-6
5.3.2.1.9 Solvent Yellow 1, C.I. 11000
e.g. Sigma Aldrich 18 636-8
5.3.2.1.10 Solvent Yellow 2, C.I. 11020
e.g. Sigma Aldrich 11 449-9
5.3.2.1.11 Solvent Yellow 3, C.I. 11160
e.g. Sigma Aldrich 12 156-8
5.3.2.1.12 Basic Red 9, C.I. 42500
e.g. Sigma Aldrich 21 559-7
5
The suppliers of the colourants mentioned in this subclause are examples of suppliers of suitable products available
commercially. This information is given for the convenience of users of this European Standard and does not constitute an
endorsement by CEN.
10
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5.3.2.1.13 Basic Violet 1, C.I. 42535
e.g. Sigma Aldrich 19 809-9
5.3.2.1.14 Basic Violet 3, C.I.42555
e.g. Sigma Aldrich 86 099-9
5.3.2.1.15 Acid Red 26, C.I.16150
e.g. Sigma Aldrich 19 976-1
5.3.2.1.16 Acid Violet 49, C.I. 42640
e.g. Sigma Aldrich S334294
5.3.2.2 Reagents and solvents
5.3.2.2.1 Tetrabutylammonium hydroxide solution, 40 % in water
5.3.2.2.2 Citric acid
5.3.2.2.3 Ammonium acetate, anhydrous
5.3.2.2.4 Acetonitrile
5.3.2.2.5 Tetrahydrofuran
5.3.2.2.6 Ethanol, absolute
5.3.2.2.7 Ammonium hydroxide, approx. 35 % (V/V)
5.3.2.2.8 Acetic acid, glacial
5.3.2.2.9 Ammonium acetate, 10 mmol/l aqueous solution, pH 3,6
Transfer (0,77 ± 0,01) g ammonium acetate (5.3.2.2.3) into a 1 000-ml volumetric flask, add 980 ml of water,
adjust the pH to 3,6 ± 0,1 with glacial acetic acid and make up to the mark with water.
5.3.2.2.10 Citrate-buffered tetrabutylammonium hydroxide solution
Transfer (13,6 ± 0,1) g tetrabutylammonium hydroxide solution (5.3.2.2.1) and (2,8 ± 0,1) g citric acid into a
1 000-ml volumetric flask, add 980 ml of water, adjust the pH to 9,0 ± 0,1 with ammonium hydroxide
(5.3.2.2.7) and make up to the mark with water.
5.3.3 Standard solutions
5.3.3.1 General
When preparing stock solutions of each colourant, purity values shall be taken into account. Store the stock
standard solutions in a refrigerator at (4 ± 2) °C.
11
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EN 71-11:2005 (E)
5.3.3.2 Stock standard solution (50 µg/ml), mix 1
Weigh, to the nearest 0,1 mg, (5 ± 1) mg of each colourant listed below into a 100-ml volumetric flask. Add
50 ml of ethanol (5.3.2.2.6) and mix carefully to dissolve. Place in an ultrasonic bath for 15 min to ensure
complete dissolution. Make up to the mark with ethanol.
Disperse Blue 1
Disperse Blue 106
Disperse Blue 124
Disperse Orange 3
Disperse Orange 37
Solvent Yellow 1
Solvent Yellow 2
Solvent Yellow 3
Basic Red 9
Basic Violet 1
Basic Violet 3
5.3.3.3 Stock standard solution (50 µg/ml), mix 2
Weigh, to the nearest 0,1 mg, (5 ± 1) mg of each colourant listed below into a 100-ml volumetric flask. Add
50 ml of ethanol and mix carefully to dissolve. Place in an ultrasonic bath for 15 min to ensure complete
dissolution. Make up to the mark with ethanol.
Disperse Blue 3
Disperse Yellow 3
Disperse Red 1
Acid Red 26
Acid Red 49
5.3.4 Apparatus
5.3.4.1 PTFE membrane filter, 0,45 µm
5.3.4.2 Ultrasonic bath
12
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5.3.4.3 Liquid chromatograph with diode-array detector
The following LC-DAD conditions for colourant analysis have been found to be suitable:
6
Column: C18, 100 Å, 5 µm, endcapped, (Luna C18(2) , or equivalent), 250 mm x 4,6 mm
6
Guard column: 2 x C18, 100 Å, 5 µm, endcapped, (Luna C18(2) , or equivalent)
Column temperature: 25 °C
Mobile phase A: Citrate-buffered tetrabutylammonium hydroxide solution (5.3.2.2.10)
Mobile phase B: Tetrahydrofuran
Mobile phase C: Acetonitrile
Gradient: see Table 3
Run time: 45 min
Flow rate 0,8 ml/min
Injection volume: 5 µl to 50 µl
Analysis time: 35 min
Wavelength range: 275 nm to 760 nm
Resolution factor: 4,8 nm
Acquisition rate: 1 spectrum/second
Table 3 – Gradient program
Time Mobile phase A Mobile phase B Mobile phase C
min % % %
0 80,0 10,0 10,0
2,50 80,0 10,0 10,0
30,0 5,0 48,0 47,0
35,0 5,0 48,0 47,0
45,0 80,0 10,0 10,0
5.3.5 Procedure
5.3.5.1 Calibration solutions
Prepare two series of colourant calibration solutions from the stock standard solutions of mix 1 (5.3.3.2) and
mix 2 (5.3.3.3) at 1 mg/l, 2 mg/l, 3 mg/l, 4 mg/l and 5 mg/l concentrations in ethanol.
5.3.5.2 Determination
Proceed to liquid chromatographic determination using the conditions described in 5.3.4.3. Inject the
calibration solutions of both mix 1 and mix 2 (5.3.5.1) and the ethanol phase obtained at 8.1.3, 8.2.1, 8.3.1,
8.4.1, 8.5.1, 8.6.1, 8.7.1, 8.8.1 or 8.9.1 of EN 71-10:2005, as appropriate.
5.3.5.3 Identification
For a positive identification, the peak purity factor should achieve a match of at least 85 %.
6
Luna C18(2) is an example of a suitable product available commercially. This information is given for the convenience of
users of this European Standard and does not constitute an endorsement by CEN of this product.
13
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EN 71-11:2005 (E)
5.3.6 Calculation of analyte concentration
Determine the concentration of a colourant in the ethanolic extract from a calibration graph produced from the
calibration solutions.
Calculate the concentration of a colourant in the sample using the following equation:
C [mg /l ]
comp, solvent
Conc [mg / kg ] = xD (2)
A
where
C is the concentration of a colourant in ethanolic extract
comp, solvent
A is the mass in grams of the test portion taken for analysis (see 8.1.3, 8.2.1, 8.3.1, 8.4.1,
8.5.1, 8.6.1, 8.7.1, 8.8.1 or 8.9.1 of EN 71-10:2005, as appropriate)
D is the dilution factor; generally 10, but see 8.5 of EN 71-10:2005 for aqueous liquid toy
material samples.
5.3.7 Limits and precision
Table 4 – Limits and precision
Substance Action limit RSD
mg/kg % at 5 mg/l
(equivalent to 10 mg/kg
in a sample)
Disperse Blue 1 10 1,8
Disperse Blue 3 10 4,9
Disperse Blue 106 10 4,4
Disperse Blue 124 10 2,2
Disperse Yellow 3 10 0,3
Disperse Orange 3 10 1,6
Disperse Orange 37 10 2,8
Disperse Red 1 10 1,6
Solvent Yellow 1 10 1,1
Solvent Yellow 2 10 1,1
Solvent Yellow 3 10 1,6
Basic Red 9 10 1,1
Basic Violet 1 10 1,5
Basic Violet 3 10 1,0
Acid Red 26 10 2,1
Acid Violet 49 10 1,4
Correlation coefficient (r): > 0,995
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SIST EN 71-11:2006
EN 71-11:2005 (E)
5.3.8 Additional information
5.3.8.1 LC-DAD spectral library
It is advisable to compile a spectral library of all colours listed in 5.3.2.1 using the processing software that is
used to operate the liquid chromatograph. Details of retention times and Lambda max of each colourant
should be recorded together with peak purity data, if available.
It has been observed that several of the colourants separated into two or more chromatographic peaks. The
colourants concerned were C.I. Acid Red 26, C.I. Disperse Blue 3, C.I. Acid Violet 49 and C.I. Basic Violet 1.
In an attempt to characterise these peaks, an LC-MS method was developed in tandem with the LC-DAD
method. The conditions used for the LC-MS analysis are detailed in Annex C.
5.3.9 Test report
The test report shall contain the following information:
a) description and identification of the product and material tested;
b) reference to this European Standard;
c) identification of colourants in the ethanolic extract of the test portion;
d) amount of each colourant identified expressed as a concentration (mg/kg) in the toy material;
e) whether a confirmation test has been carried out and, if so, the technique used and its result;
f) any deviations from the test procedure specified;
g) date of test.
5.4 Primary aromatic amines
5.4.1 Principle
Aromatic amines are determined in extracts of toy materials by gas chromatography with mass spectrometry
detection (GC-MS) using the external standard method of calibration combined with appropriate internal
standards.
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SIST EN 71-11:2006
EN 71-11:2005 (E)
5.4.2 Standards, reagents and solvents
5.4.2.1 Standards
5.4.2.1.1 Benzidine
5.4.2.1.2 Aniline
5.4.2.1.3 2-Naphthylamine
5.4.2.1.4 3,3’-Dichlorobenzidine
5.4.2.1.5 3,3’-Dimethoxybenzidine
5.4.2.1.6 3,3’-Dimethylbenzidine
5.4.2.1.7 2-Methoxyaniline
5.4.2.1.8 o-Toluidine
5.4.2.1.9 4-Chloroaniline
5.4.2.2 Reagents and solvents
Acetonitrile
5.4.2.2.1
tert-Butyl methyl ether
5.4.2.2.2
5.4.2.2.3 n-Hexane
7
5.4.2.2.4 Chromabond XTR (Porous granulated kieselguhr)
5.4.2.3 Stock standard solution (100 mg/l)
Weigh, to the nearest 0,1 mg, (10 ± 1) mg of each aromatic amine (5.4.2.1) into a 100-ml volumetric flask.
Add 25 ml of acetonitrile (5.4.2.2.1) and mix carefully to dissolve. Place in an ultrasonic bath for 10 min to
ensure complete dissolution. Make up to the mark with acetonitrile.
The stability of the mixed stock standard solution should be checked regularly. It should be stable for up to 6
months when stored in the dark at (4 ± 2) °C.
7
Chromabond XTR is an example of a suitable product available commercially. This information is given for the
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
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SIST EN 71-11:2006
EN 71-11:2005 (E)
5.4.3 Apparatus
5.4.3.1 Ultrasonic bath
5.4.3.2 Vortex® shaker
5.4.3.3 Centrifuge
5.4.3.4 Gas chromatograph with mass spectrometer detector
Ensure that the gas chromatograph has been fully cleaned before analysis, as the determination of aromatic
amines by this technique is affected by other contaminants. It is recommended that the inlet sleeve is
deactivated and an amine-specific column used.
The following GC-MS conditions for primary aromatic amine determination have been found to be suitable:
Injector:
Mode: Splitless 0,5 min
C
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