DIN EN 16344-2013 Cosmetics - Analysis of cosmetic products - Screening for UV-filters in cosmetic products and quantitative determination of 10 UV-filters by HPLC German version E.pdf

1、November 2013 Translation by DIN-Sprachendienst.English price group 13No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).I

2、CS 71.100.70!%*,d“2070965www.din.deDDIN EN 16344Cosmetics Analysis of cosmetic products Screening for UV-filters in cosmetic products and quantitativedetermination of 10 UV-filters by HPLC;English version EN 16344:2013,English translation of DIN EN 16344:2013-11Kosmetische Mittel Untersuchung von ko

3、smetischen Mitteln Screening und quantitative Bestimmung von 10 UV-Filtern in Sonnenschutzmitteln,HPLC-Verfahren;Englische Fassung EN 16344:2013,Englische bersetzung von DIN EN 16344:2013-11Cosmtiques Analyse des produits cosmtiques Dtection des filtres UV dans les produits cosmtiques et dterminatio

4、n quantitative de 10filtres UV par CLHP;Version anglaise EN 16344:2013,Traduction anglaise de DIN EN 16344:2013-11www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 27 pages 10.13 DIN EN 16344:2013-11 2 A comma is used as the decimal marke

5、r. National foreword This document (EN 16344:2013) has been prepared by Technical Committee CEN/TC 392 “Cosmetics” (Secretariat: AFNOR, France). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Produc

6、ts Standards Committee), Working Committee NA 057-07-01 AA Kosmetische Mittel. EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16344 July 2013 ICS 71.100.70 English Version Cosmetics - Analysis of cosmetic products - Screening for UV-filters in cosmetic products and quantitative determination o

7、f 10 UV-filters by HPLC. Cosmtiques - Analyse des produits cosmtiques - Dtection des filtres UV dans les produits cosmtiques et dtermination quantitative de 10 filtres UV par CLHP Kosmetische Mittel - Untersuchung von kosmetischen Mitteln - Screening und quantitative Bestimmung von 10 UV-Filtern in

8、Sonnenschutzmitteln, HPLC-Verfahren This European Standard was approved by CEN on 29 May 2013. 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-da

9、te 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

10、 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, For

11、mer 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 COMIT

12、 EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16344:2013: EEN 16344:2013 (E) 2 Contents Page Foreword . 3 Introduc

13、tion 4 1 Scope 5 2 Terms and definitions 5 3 Principle 5 4 Reagents . 6 5 Apparatus and equipment . 11 6 Sampling . 11 7 Procedure . 12 8 Evaluations . 14 9 Test report 15 Annex A (informative) Results of the inter-laboratory test 16 Annex B (informative) Sample Chromatogram 1 . 22 Annex C (informat

14、ive) Sample chromatogram 2 24 Bibliography 25 DIN EN 16344:2013-11 EN 16344:2013 (E) 3 Foreword This document (EN 16344:2013) has been prepared by Technical Committee CEN/TC 392 “Cosmetics”, the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national sta

15、ndard, either by publication of an identical text or by endorsement, at the latest by January 2014, and conflicting national standards shall be withdrawn at the latest by January 2014. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent right

16、s. 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 organisations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cy

17、prus, 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 Unit

18、ed Kingdom. DIN EN 16344:2013-11 EN 16344:2013 (E) 4 Introduction Reference is made to the relevant annex of the Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products: Annex VI List of UV-filters allowed in cosmetic products. DIN EN 16344

19、:2013-11 EN 16344:2013 (E) 5 1 Scope This European Standard specifies a multi-screening method using reversed-phase HPLC for the detection of UV-filters listed in the cosmetic regulations. The method is applicable for the quantitative determination of 10 UV-filters, which are mainly used in emulsion

20、-based cosmetic products and sunscreen sprays particularly with regard to the maximum concentration listed in the cosmetic regulation. Other analytical methods for the qualification and quantification of UV-filters may be used if they lead to comparable results. 2 Terms and definitions For the purpo

21、ses of this document, the following terms and definitions apply. 2.1 UV-filters organic chemical compounds that absorb ultraviolet light and inorganic substances that reflect, scatter and absorb UV light Note 1 to entry: The UV-filters and UV-absorber of this method are only organic chemical compoun

22、ds and are used in sunscreen products to protect the skin against UV radiation. 3 Principle The UV-filters are extracted with an acetone/methanol mixture. For the qualitative detection of the listed UV-filters and the quantitative determination of the 10 validated UV-filter reversed phase HPLC with

23、UV (DAD) detection is used. The method is also applicable for the quantification of the other listed UV-filters after proper validation. Quantitative determination of samples containing the following UV-filters require the use of additional extraction methods and determinations: Terephthalylidene Di

24、camphor Sulfonic Acid (TDSA) and Disodium Phenyl Dibenzimidazole Tetrasulfonate (DPDT) are additionally extracted with methanolic-aqueous sodium hydroxide solution. Methylene Bis-benzotriazolyl Tetramethylbutylphenol (MBBT) and Bis-Ethylhexyloxyphenol Methoxyphenyl Triazine (BEMT) are additionally e

25、xtracted with a mixture of tetrahydrofuran/acetone. In the case of an unsatisfactory peak shape, Butyl Methoxydibenzoylmethane (BMDM) is additionally extracted with a mixture of acetone/methanol/EDTA. The quantitative determination is made by means of RP-HPLC with UV (DAD). The UV-spectra are compar

26、ed with the reference spectra in a database. The concentration of each UV-filter determined in accordance with this method is reported in g/100 g. This method has been tested in an inter-laboratory test on specific cosmetic matrix (q.v. Annex A). The user should verify the performance of the method

27、in their laboratory for each different matrix and pay particular attention to the recommended quality control elements. DIN EN 16344:2013-11 EN 16344:2013 (E) 6 4 Reagents 4.1 General If not otherwise specified, analytical-grade chemicals shall be used. Water shall be distilled or of a corresponding

28、 purity. “Solution“ shall be understood as an aqueous solution unless otherwise specified. 4.2 Methanol (MeOH), HPLC grade. 4.3 Acetone, HPLC grade. 4.4 Tetrahydrofuran (THF), HPLC grade. 4.5 Ammonia solution, mass fraction = 25 g/100 g. 4.6 Sodium hydroxide solution, molar concentration c = 1 mol/l

29、. 4.7 Ethylenediaminetetraacetic acid (EDTA) disodium salt dihydrate (Na2EDTA 2H2O, CAS 6381-92-6, purity 99 %). 4.8 EDTA solution Weigh 1,8 g of EDTA disodium salt dihydrate (4.7) into a 100 ml volumetric flask and fill up to the calibration mark with water. 4.9 Ethanol, HPLC grade. 4.10 Lauryl Tri

30、methyl Ammonium Bromide (LTAB, synonym: dodecyltrimethylammonium bromide. CAS 1119-94-4), if possible HPLC quality (purity 98 %). 4.11 Ammonium bromide (CAS 12124-97-9, purity 99 %). 4.12 Reference substances DIN EN 16344:2013-11 EN 16344:2013 (E) 7 Table 1 Polar UV-filters (calibration solution in

31、methanol) EUaAbbrev. INCIband other common names 4.12.1 A2 CBM Camphor Benzalkonium Methosulfate, CAS 52793-97-2 4.12.2 A6 PBSA Phenylbenzimidazole Sulfonic Acid (2-phenylbenzimidazole-5-sulfonic acid), CAS 27503-81-7 4.12.3 A7 TDSA Terephthalylidene Dicamphor Sulfonic Acid, CAS 90457-82-2, present

32、as triethanolamine salt (molecular weight m = 860 g/mol), free acid (molecular weight m = 562 g/mol) 4.12.4 A22 B-4/5 Benzophenone-4/5 (2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, Sulisobenzone), CAS 4065-45-6 4.12.5 A24 DPDT Disodium Phenyl Dibenzimidazole Tetrasulfonate, CAS 180898-37-7 4.12.

33、6 A28 DHHB Diethylamino Hydroxybenzoyl Hexyl Benzoate, CAS 302776-68-7 aEU = serial number in accordance with Annex VI of (EC) No 1223/2009. bINCI = International Nomenclature of Cosmetic Ingredients. Table 2 Medium polar UV-filters (calibration solution in methanol acetone (1:1) EUaAbbrev. INCIband

34、 other common names 4.12.7 A4 B-3 Benzophenone-3 (oxybenzonum, 2-hydroxy-4-methoxy-benzophenone), CAS 131-57-7 4.12.8 A10 OC Octocrylene (2-ethylhexyl-2-cyano-3,3-diphenylacrylate), CAS 6197-30-4 4.12.9 A12 EHMC Ethylhexyl Methoxycinnamate (octylmethoxycinnamate), CAS 5466-77-3 4.12.10 A14 IMC Isoam

35、yl p-Methoxycinnamate, CAS 71617-10-2 4.12.11 A18 MBC 4-Methylbenzylidene Camphor (3-(4-methylbenzylidene)-dl-camphor), CAS 36861-47-9 4.12.12 A19 3-BC 3-Benzylidene Camphor, CAS 15087-24-8 4.12.13 A21 EHDP Ethylhexyl Dimethyl PABA (2-ethylhexyl-4-dimethylaminobenzoate), CAS 21245-02-3 aEU = serial

36、number in accordance with Annex VI of (EC) No 1223/2009. bINCI = International Nomenclature of Cosmetic Ingredients. Table 3 Non polar UV-filters (calibration solution in THF) EUaAbbrev. INCIband other common names 4.12.14 A3 HMS Homosalate (3,3,5-trimethylcyclohexylsalicylate), CAS 118-56-9 4.12.15

37、 A8 BMDM Butyl Methoxydibenzoylmethane (4-tert-butyl-4-methoxydibenzoylmethane), CAS 70356-09-1 4.12.16 A15 EHT Ethylhexyl Triazone (octyltriazone), CAS 88122-99-0 4.12.17 A16 DTS Drometrizole Trisiloxane (2-benzotriazole-2-yl-methylphenol trisiloxane), CAS 155633-54-8 4.12.18 A17 DEBT Diethylhexyl

38、Butamido Triazone, CAS 154702-15-5 4.12.19 A20 EHS Ethylhexyl Salicylate (2-ethylhexylsalicylate), CAS 118-60-5 4.12.20 A23 MBBT Methylene Bis-Benzotriazol Tetramethylbutylphenol, CAS 103597-45-1 4.12.21 A25 BEMT Bis-Ethylhexyloxyphenol Methoxyphenyl Triazine (anisotriazine), CAS 187393-00-6 aEU = s

39、erial number in accordance with Annex VI of (EC) No 1223/2009. bINCI = International Nomenclature of Cosmetic Ingredients. DIN EN 16344:2013-11 EN 16344:2013 (E) 8 Table 4 Other UV-filter or UV absorber (not listed in Annex VI of the (EC) No 1223/2009) Abbrev. INCIband other common names 4.12.22 MA

40、Menthyl Anthranilate, CAS 134-09-8 4.12.23 PABA PABA (4-aminobenzoic acid), CAS 150-13-0 4.12.24 B-1 Benzophenone-1 (2,4-dihydroxybenzophenone), CAS 131-56-6 4.12.25 B-2 Benzophenone-2 (2,2,4,4-tetrahydroxybenzophenone), CAS 131-55-5 4.12.26 DPLT Dimethyl-PABAmidopropyl Laurdimonium Tosylate, CAS 15

41、6679-41-3 4.12.27 B-6 Benzophenone-6 (2,2-dihydroxy-4,4-dimethoxybenzophenone), CAS 131-54-4 4.12.28 B-8 Benzophenone-8 (2,2-dihydroxy-4-methoxybenzophenone, dioxybenzone),CAS 131-53-3 4.12.29 B-9 Benzophenone-9 (disodium 3,3-carbonylbis4-hydroxy-6-methoxybenzenesulphonate), CAS 76656-36-5 4.12.30 B

42、-10 Benzophenone-10 (2-hydroxy-4-methoxy-4-methylbenzophenone), CAS 1641-17-4 4.12.31 SA Sodium Salicylate, CAS 54-21-7 bINCI = International Nomenclature of Cosmetic Ingredients. NOTE The following UV-filters listed in Annex VI of the Regulation (EC) No 1223/2009 are not part of this method: A 9: 3

43、-(4-sulfo)-benzylidene-bornan-2-one, CAS 56039-58-8 A 11: polymer of N-(2(and 4)-(2-oxoborn-3-ylidenemethyl)benzyl)acrylamid, CAS 113783-61-2 (the substances are no longer commercially available) A 13: ethoxylated ethyl-4-aminobenzoate, CAS 116242-27-4 (can be determined only qualitatively) A 26: di

44、methicodiethylbenzalmalonate, CAS 207574-74-1 (cannot be determined using this standard) 4.13 Extraction solution 4.13.1 Acetone/methanol mixture (for the preparation of the reference solutions and for extraction). Mix 500 ml of acetone (4.3) and 500 ml of methanol (4.2) in a 1 000 ml conical flask.

45、 4.13.2 Acetone/tetrahydrofuran mixture (for extraction of non-polar UV-filters). Mix 500 ml of acetone (4.3) and 500 ml of tetrahydrofuran (4.4) in a 1 000 ml conical flask. 4.13.3 Methanolic-aqueous sodium hydroxide solution (for extraction of polar UV-filters). Mix 800 ml of methanol (4.2) and 20

46、0 ml of water and 10 ml of sodium hydroxide solution (4.6) in a 1 000 ml conical flask. 4.13.4 Acetone/methanol/EDTA mixture (for extraction of BMDM-containing samples with unsatisfactory peak shape). Add 1,0 ml of EDTA solution (4.8) to 200 ml of acetone/methanol mixture (4.13.1) in a 200 ml conica

47、l flask. As the solution is supersaturated with regard to EDTA it shall be used on the same day. DIN EN 16344:2013-11 EN 16344:2013 (E) 9 4.14 Reference solutions 4.14.1 Stock solutions The stock solutions are prepared in accordance with Table 5, Table 6 and Table 7. For each substance, weigh the in

48、itial weights with an accuracy of 0,1 mg to the mentioned volume. One stock solution is prepared per substance. For the initial weight, the purity of the substances shall be taken into account and if applicable be converted to an initial weight of 4 mg/ml of “active substance“ in each case. The stock solutions can be stored for at least one month in a refrigerator between 2 C and 8 C in the absence of light. Table 5 Scheme for preparing the stock solutions of polar UV-filters Polar UV-filter Initial we