DIN EN 16343-2013 Cosmetics - Analysis of cosmetic products - Determination of 3-iodo-2-propynyl butylcarbamate (IPBC) in cosmetic preparations LC-MS methods German version EN 1634.pdf

1、August 2013 Translation by DIN-Sprachendienst.English price group 8No 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).ICS

2、71.100.70!%fA“2046730www.din.deDDIN EN 16343Cosmetics Analysis of cosmetic products Determination of 3-iodo-2-propynyl butylcarbamate (IPBC) in cosmeticpreparations, LC-MS methods;English version EN 16343:2013,English translation of DIN EN 16343:2013-08Kosmetische Mittel Untersuchung von kosmetische

3、n Mitteln Bestimmung von 3-Iod-2-propinylbutylcarbamat (IPBC) in kosmetischen Mitteln,LC-MS-Verfahren;Englische Fassung EN 16343:2013,Englische bersetzung von DIN EN 16343:2013-08Cosmtiques Analyse des produits cosmtiques Dtermination quantitative du carbamate 3-iodo-2-propynylbutyle (IPBC),mthodes

4、CL-SM;Version anglaise EN 16343:2013,Traduction anglaise de DIN EN 16343:2013-08www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 11 pages 07.13 DIN EN 16343:2013-08 2 A comma is used as the decimal marker. National foreword This document

5、 (EN 16343:2013) 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 Products Standards Committee), Working C

6、ommittee NA 057-07-01 AA Kosmetische Mittel. EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16343 May 2013 ICS 71.100.70 English Version Cosmetics - Analysis of cosmetic products - Determination of 3-iodo-2-propynyl butylcarbamate (IPBC) in cosmetic preparations, LC-MS methods Cosmtiques - Ana

7、lyse des produits cosmtiques - Dtermination quantitative du carbamate 3-iodo-2-propynylbutyle (IPBC), mthodes CL-SM Kosmetische Mittel - Untersuchung von kosmetischen Mitteln - Bestimmung von 3-Iod-2-propinylbutylcarbamat (IPBC) in kosmetischen Mitteln, LC-MS-Verfahren This European Standard was app

8、roved by CEN on 25 April 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-date lists and bibliographical references concerning such national

9、 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

10、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, Hun

11、gary, 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 EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Managem

12、ent 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 16343:2013: EEN 16343:2013 (E) 2 Contents Page Foreword . 3 1 Scope 4 2 Principle 4 3 Reagents . 4 4 Apparatus and equipment . 5 5 P

13、rocedure . 5 6 Evaluation . 7 7 Test report 7 Annex A (informative) Results of the inter-laboratory test . 8 Bibliography 9 DIN EN 16343:2013-08 EN 16343:2013 (E) 3 Foreword This document (EN 16343:2013) has been prepared by Technical Committee CEN/TC 392 “Cosmetics”, the secretariat of which is hel

14、d by AFNOR. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by November 2013, and conflicting national standards shall be withdrawn at the latest by November 2013. Attention is drawn to the possibili

15、ty 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 bo

16、und 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, Portug

17、al, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 16343:2013-08 EN 16343:2013 (E) 4 1 Scope This European Standard specifies a method for the quantitative determination of 3-iodo-2-propynyl butylcarbamate (IPBC) in the concentration range from 0,005 g

18、/100 g to 0,1 g/100 g - Annex V No. 56 in Regulation (EC) No 1223/2009 on cosmetic products. 2 Principle IPBC is extracted from the cosmetic preparation using methanol. IPBC present in the sample extract is separated using reverse phase HPLC with mass specific detection (LC-MS or LC-MS/MS). Quantita

19、tive determination of IPBC is made using the external standard method of calibration or standard addition. 3 Reagents 3.1 General If not otherwise specified, analytical-grade chemicals shall be used; water shall be distilled or of a suitable purity. “Solution” shall be understood as an aqueous solut

20、ion unless otherwise specified. 3.2 Iodopropynyl butylcarbamat, CAS number: 55406-53-6 (Supplier : Sigma-Aldrich1)(521949), Dr. Ehrenstorfer GmbH1)(C 14335000). 3.3 Methanol, HPLC grade, CAS number: 67-56-1. 3.4 Formic acid, CAS number: 64-18-6. 3.5 Tetrahydrofuran (THF), CAS number: 109-99-9. 3.6 P

21、ropan-2-ol, CAS number: 67-63-0. 3.7 Eluents 3.7.1 Eluent A, 1 ml of formic acid (3.4) is mixed with 1 000 ml of water. 3.7.2 Eluent B, Methanol (3.3). 3.8 IPBC stock solution, = 1 mg/ml. Weigh approximately 0,05 g of IPBC (3.2) into a 50-ml-volumetric flask. Firstly, dissolve in a small amount of m

22、ethanol (3.3) and then fill up to the calibration mark with methanol. This solution has a shelf life of 8 weeks if it is stored in a refrigerator. 3.9 Calibration solutions (standard solutions) 5,0 ml of the stock solution (3.8) is transferred into a 50-ml-volumetric flask and filled to the calibrat

23、ion mark with methanol (3.3), ( = 0,1 mg/ml or 100 g/ml). From this solution, at least 5 solutions are prepared by dilution to obtain IPBC concentrations of = 0,05 g/ml to = 1,0 g/ml. These solutions have a shelf life of 8 weeks if they are stored in a cool place. Examples of dilutions are given in

24、Table 1. 1) This is an example of a suitable product available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. DIN EN

25、 16343:2013-08 EN 16343:2013 (E) 5 Table 1 Calibration solutions No. Preparation Dilution Concentration g/ml 3.9 5 ml 3.8 filled up to 50 ml 1:10 100 3.10 10 ml 3.9 filled up to 100 ml 1:100 10 3.11 10 ml 3.10 filled up to 100 ml 1:1000 1 3.12 4 ml 3.10 filled up to 50 ml 1:1250 0,8 3.13 5 ml 3.10 f

26、illed up to 100 ml 1:2000 0,5 3.14 10 ml 3.11 filled up to 50 ml 1:5000 0,2 3.15 5 ml 3.11 filled up to 50 ml 1:10000 0,1 4 Apparatus and equipment 4.1 Standard laboratory equipment 4.2 Membrane filter, in the form of a disposable syringe filter, pore width: 0,2 m2). 4.3 High-performance liquid chro

27、matograph, suitable for gradient elution with mass-specific detector. 4.4 Analytical separation column, the following parameters have proved useful: RP 18 Phase, 5 m, 150 mm 2 mm, e.g. Zorbax1), Spherisorb1), Phenomenex-Luna1)or equivalent. If a precolumn is used, it shall have the same analytical p

28、roperties as the separation column. 5 Procedure 5.1 Sample preparation Weigh approximately 200 mg of the sample to the nearest 0,1 mg into a 20-ml-volumetric flask (alternatively a 50-ml-volumetric flask). Add 1,5 ml of THF (3.5) and shake. Add 10 ml of methanol (3.3), and allow to dissolve or suspe

29、nd for 5 min in the ultrasonic bath at room temperature. Allow to cool to room temperature and fill to the calibration mark with methanol (3.3). The sample solution is diluted with methanol (3.3) to a ratio of 1:10, filtered through a membrane filter (4.2) and then analysed using LC-MS or LC-MS/MS.

30、For poorly soluble or suspendable matrices it is recommended to partially dissolve the sample by adding 2 ml of 2-propanol (3.6) instead of THF (3.5) or to stir it with a magnetic stirrer for 30 min prior to the treatment in the ultrasonic bath. 5.2 Liquid chromatography (LC) conditions When using t

31、he apparatus (4.3) and column (4.4), the following conditions have shown to be useful (see Table 2): 2) The ring test was performed using 0,2 m filter. DIN EN 16343:2013-08 EN 16343:2013 (E) 6 Table 2 Gradient programme Time min Fraction eluent A % Fraction eluent B % 0 85 15 8 10 90 12 10 90 13 85

32、15 25 85 15 Column: RP 18 Phase, 5 m, 150 mm 2 mm Injection volume: 1 l to 10 l Flow rate: 0,2 ml/min Column oven temperature: 25 C 5.3 Detection 5.3.1 General The detection and quantitative determination can be performed by evaluating the mass traces of IPBC or by evaluating the fragment ions. To a

33、void false low results by adduct formation occurring in the MRM mode of the Electrospray (ESI) method, the APCI (Atmospheric Pressure Chemical Ionisation) method should be used for ionisation. 5.3.2 MS detection in Selected Ion Monitoring (SIM) mode Mass traces: m/z 282 M+H+and m/z 304 M+Na+. Evalua

34、tion is based on the total ion current (from the sum of the two masses). 5.3.3 MS detection in Multiple Reaction Monitoring (MRM) mode Protonated molecule ion: 282 M+H+Fragment ion 1: 57 Fragment ion 2: 165 Evaluation is based on the most sensitive fragment ion. Since the analyte may form adducts wi

35、th sodium ions, false low results can be obtained if larger amounts of sodium ions are present in the sample. If ESI ionisation is used, standard addition should therefore be performed using the MRM mode so as to ensure reliable quantification results. When performing the standard addition, the cont

36、ent of the added IPBC should not exceed the content expected in the sample. DIN EN 16343:2013-08 EN 16343:2013 (E) 7 6 Evaluation 6.1 Identification and quantitative determination The IPBC is identified by comparing the retention times of the sample with those of calibration solutions. The quantitat

37、ive determination of the analyte is performed based on a calibration function or the standard addition. The calibration solutions are chromatographed in accordance with the conditions given in 5.2. The IPBC concentration is calculated from the calibration by linear regression on the basis of the obt

38、ained peak area. 6.2 Calculation The preservative content w in g/100 g, with respect to the sample, is calculated using the following formula: 10001000100=mFVcw (1) where w is the IPBC content, in g/100 g; c is the IPBC concentration in the sample solution, in g/ml, determined from the calibration f

39、unction; m is the initial weight of the sample, in g; V is the volume of the sample measurement solution, in ml; F is the dilution factor, if required. The result is given in g/100 g, rounded to three decimal places. 7 Test report The test report shall contain the following data: a) information nece

40、ssary for the identification of the sample (type, origin and designation of the sample); b) a reference to this European Standard; c) name of the laboratory performing the test; d) the date and type of sampling procedure (if known); e) the date of receipt and date of analysis; f) the date of test; g

41、) the test results and the units in which they have been expressed; h) justification of any deviations from this official method; i) operations not specified in the method or regarded as optional, which might have affected the results. DIN EN 16343:2013-08 EN 16343:2013 (E) 8 Annex A (informative) R

42、esults of the inter-laboratory test This method has been developed by the working group “Cosmetics” of the German Federal Office of Consumer Protection and Food Safety (BVL) for the purpose of implementing Section 64 of the Food and Feed Code (LFGB). It has been tested in an inter-laboratory test wi

43、th a total of 13 participants. The following statistical data presented in Table A.1 have been determined for shower gel and cream in an inter-laboratory test with 13 participating laboratories using external quantification. Table A.1 Statistical data of the inter-laboratory test Parameter LC-MS met

44、hod IPBC content, in g/100 g Shower gel Cream Number of participating laboratories 13 13 Number of outliers 3 2 Number of laboratories after elimination of the outliers 10 11 Mean value x , g/100 g 0,013 0,019 Repeatability r, g/100 g 0,003 0,003 Repeatability standard deviation sr, g/100 g 0,001 0,

45、001 Reproducibility R, g/100 g 0,006 0,005 Reproducibility standard deviation sR, g/100 g 0,002 0,002 Recovery, % 101,4 103,7 DIN EN 16343:2013-08 EN 16343:2013 (E) 9 Bibliography 1 Frauen M, Steinhart H, Rapp C, Hintze U. (2001): Rapid quantification of iodopropynyl butylcarbamate as the preservative in cosmetic formulations using high-performance liquid chromatography-electrospray mass spectrometry. J Pharm Biomed Anal. 25 (5-6):965-70 DIN EN 16343:2013-08